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Purpose: Vibrio vulnificus (V. Vulnificus) infection is characterized by rapid onset, aggressive progression, and challenging treatment. Bacterial resistance poses a significant challenge for clinical anti-infection treatment and is thus the subject of research. Enhancing host infection tolerance represents a novel infection prevention strategy to improve patient survival. Our team initially identified cytochrome P4501A1 (CYP1A1) as an important target owing to its negative modulation of the body's infection tolerance. This study explored the superior effects of the CYP1A1 inhibitor bergamottin compared to antibiotic combination therapy on the survival of mice infected with multidrug-resistant V. Vulnificus and the protection of their vital organs., Methods: An increasing concentration gradient method was used to induce multidrug-resistant V. Vulnificus development. We established a lethal infection model in C57BL/6J male mice and evaluated the effect of bergamottin on mouse survival. A mild infection model was established in C57BL/6J male mice, and the serum levels of creatinine, urea nitrogen, aspartate aminotransferase, and alanine aminotransferase were determined using enzyme-linked immunosorbent assay to evaluate the effect of bergamottin on liver and kidney function. The morphological changes induced in the presence of bergamottin in mouse organs were evaluated by hematoxylin and eosin staining of liver and kidney tissues. The bacterial growth curve and organ load determination were used to evaluate whether bergamottin has a direct antibacterial effect on multidrug-resistant V. Vulnificus. Quantification of inflammatory factors in serum by enzyme-linked immunosorbent assay and the expression levels of inflammatory factors in liver and kidney tissues by real-time quantitative polymerase chain reaction were performed to evaluate the effect of bergamottin on inflammatory factor levels. Western blot analysis of IκBα, phosphorylated IκBα, p65, and phosphorylated p65 protein expression in liver and kidney tissues and in human hepatocellular carcinomas-2 and human kidney-2 cell lines was used to evaluate the effect of bergamottin on the nuclear factor kappa-B signaling pathway. One-way ANOVA and Kaplan-Meier analysis were used for statistical analysis., Results: In mice infected with multidrug-resistant V. Vulnificus, bergamottin prolonged survival (p = 0.014), reduced the serum creatinine (p = 0.002), urea nitrogen (p = 0.030), aspartate aminotransferase (p = 0.029), and alanine aminotransferase (p = 0.003) levels, and protected the cellular morphology of liver and kidney tissues. Bergamottin inhibited interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in serum (IL-1β: p = 0.010, IL-6: p = 0.029, TNF-α: p = 0.025) and inhibited the protein expression of the inflammatory factors IL-1β, IL-6, TNF-α in liver (IL-1β: p = 0.010, IL-6: p = 0.011, TNF-α: p = 0.037) and kidney (IL-1β: p = 0.016, IL-6: p = 0.011, TNF-α: p = 0.008) tissues. Bergamottin did not affect the proliferation of multidrug-resistant V. Vulnificus or the bacterial load in the mouse peritoneal lavage fluid (p = 0.225), liver (p = 0.186), or kidney (p = 0.637)., Conclusion: Bergamottin enhances the tolerance of mice to multidrug-resistant V. Vulnificus infection. This study can serve as a reference and guide the development of novel clinical treatment strategies for V. Vulnificus., (Copyright © 2024 Chinese Medical Association. Production and hosting by Elsevier B.V. All rights reserved.)

Chuanxiong Rhizoma is a well-known Sichuan-specific herbal medicine. Its original plant, Ligusticum chuanxiong, has been cultivated asexually for a long time. L. chuanxiong has sexual reproductive disorders, which restricts its germplasm innovation. However, there is little research on the reproductive system of L. chuanxiong. This study is based on a comparative anatomical research approach, using morphological dissection, paraffin sectioning, staining and compression, and combined with scanning electron microscopy technology, to observe and compare the flowers, fruits, and seeds at various stages of reproductive growth of L. chuanxiong and its wild relative L. sinense. The results showed that the meiosis of pollen mother cells is abnormal in L. chuanxiong anthers, and the size and number of microspores are uneven and inconsistent in the tetrad stage. tapetum cells are not completely degenerated during anther development. During the pollen ripening stage, there are fine cracks in the anther wall, while most anthers could not release pollen normally. The surface of mature pollen grains is concave and partially deformed, and the pollens are all inactive and cannot germinate in vitro. The starch, polysaccharides, and lipids in the pollen were insufficient. The filaments of L. chuanxiong are short at the flowering stage and recurved downward. Double-hanging fruits were observed in the fruiting stage, being wrinkled; with shriveled seeds. Compared with L. sinense at the same stage, the anthers of L. sinense developed normally, and the pollen grains are vigorous and can germinate in vitro. The double-hanging fruits of L. sinense are full and normal; at the flowering period, the filaments are long and erect, significantly higher than the stigma. Mature blastocysts are visible in the ovary of both L. chuanxiong and L. sinense, and there is no significant difference in stigmas. The conclusion is that during the development of L. chuanxiong stamens, the meiosis of pollen mother cells is abnormal, and tetrad, tapetum, filament and other pollen structures develop abnormally. L. chuanxiong has the characteristic of male infertility, which is an important reason for its sexual reproductive disorders.

Background: Arbuscular mycorrhizal fungi (AMF) form symbiotic relationships with various terrestrial plants and have attracted considerable interest as biofertilizers for improving the quality and yield of medicinal plants. Despite the widespread distribution of AMFs in Salvia miltiorrhiza Bunge's roots, research on the impact of multiple AMFs on biomass and active ingredient accumulations has not been conducted. In this study, the effects of five native AMFs (Glomus formosanum, Septoglomus constrictum, Rhizophagus manihotis, Acaulospora laevis, and Ambispora gerdemannii) and twenty-six communities on the root biomass and active ingredient concentrations of S. miltiorrhiza were assessed using the total factor design method., Results: Thirty-one treatment groups formed symbiotic relationships with S. miltiorrhiza based on the pot culture results, and the colonization rate ranged from 54.83% to 89.97%. AMF communities had higher colonization rates and total phenolic acid concentration than single AMF, and communities also appeared to have higher root fresh weight, dry weight, and total phenolic acid concentration than single inoculations. As AMF richness increased, there was a rising trend in root biomass and total tanshinone accumulations (ATTS), while total phenolic acid accumulations (ATP) showed a decreasing trend. This suggests that plant productivity was influenced by the AMF richness, with higher inoculation benefits observed when the communities contained three or four AMFs. Additionally, the affinities of AMF members were also connected to plant productivity. The inoculation effect of closely related AMFs within the same family, such as G. formosanum, S. constrictum, and R. manihotis, consistently yielded lower than that of mono-inoculation when any combinations were applied. The co-inoculation of S. miltiorrhiza with nearby or distant AMFs from two families, such as G. formosanum, R. manihotis, and Ac. laevis or Am. gerdemannii resulted in an increase of ATP and ATTS by more than 50%. AMF communities appear to be more beneficial to the yield of bioactive constituents than the single AMF, but overall community inoculation effects are related to the composition of AMFs and the relationship between members., Conclusion: This study reveals that the AMF community has great potential to improve the productivity and the accumulation of bioactive constituents in S. miltiorrhiza, indicating that it is an effective way to achieve sustainable agricultural development through using the AMF community., (© 2023. The Author(s).)

Alzheimer's disease (AD) is a neurodegenerative disorder with cognitive impairment and abnormal mental behaviour. There is currently no effective cure. The development of early diagnostic markers and the mining of potential therapeutic targets are one of the important strategies. This study aimed to explore potential biomarkers or therapeutic targets related to AD in the hippocampus and prefrontal cortex, two brain regions highly related to AD. Differentially expressed genes and miRNAs between AD patients and healthy controls were obtained from the Gene Expression Omnibus database. The mRNA-miRNA network was constructed and key genes involved in AD were screened out by protein-protein interaction analysis, and were subsequently verified by independent datasets and qPCR in an AD mouse model. Our findings showed that six hub genes including CALN1, TRPM7, ATR, SOCS3, MOB3A and OGDH were believed to be involved in the pathogenesis of AD. Western blot analysis further determined that CALN1, ATR and OGDH were the possible biomarkers and therapeutic targets for AD. In addition, 6 possible miRNAs biomarkers have also been verified by qPCR on AD animal models. Our findings may benefit clinical diagnosis and early prevention of AD., (© 2022 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Introduction: As the main manifestation of gallstone disease, biliary colic (BC) is an episodic attack that brings patients severe pain in the right upper abdominal quadrant. Although acupuncture has been documented with significance to lead to pain relief, the immediate analgesia of acupuncture for BC still needs to be verified, and the underlying mechanism has yet to be covered. Therefore, this trial aims first to verify the immediate pain-alleviation characteristic of acupuncture for BC, then to explore its influence on the peripheral sensitised acupoint and central brain activity., Methods and Analysis: This is a randomised controlled, paralleled clinical trial, with patients and outcome assessors blinded. Seventy-two patients with gallbladder stone disease presenting with BC will be randomised into a verum acupuncture group and the sham acupuncture group. Both groups will receive one session of immediate acupuncture treatment. Improvements in patients' BC will be evaluated by the Numeric Rating Scale, and the pain threshold of acupoints will also be detected before and after treatment. During treatment, brain neural activity will be monitored with functional near-infrared spectroscopy (fNIRS), and the needle sensation will be rated. Clinical and fNIRS data will be analysed, respectively, to validate the acupuncture effect, and correlation analysis will be conducted to investigate the relationship between pain relief and peripheral-cerebral functional changes., Ethics and Dissemination: This trial has been approved by the institutional review boards and ethics committees of the First Teaching Hospital of Chengdu University of Traditional Chinese Medicine, with the ethical approval identifier 2019 KL-029, and the institutional review boards and ethics committees of the First People's Hospital of Longquanyi District, with the ethical approval identifier AF-KY-2020071. The results of this trial will be disseminated through peer-reviewed publications and conference abstracts or posters., Trial Registration Number: CTR2000034432., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

The study is aimed to reveal the fluctuation of the inorganic elements in the rhizosphere soil of Ligusticum chuanxiong during their whole growth period, and explore the relationship between that fluctuation and the formation of radial striations character in the rhizomes. During the cultivation period of L. chuanxiong, the rhizosphere soil samples were taken regularly, and the content of 26 inorganic elements were determined by X-ray fluorescence spectrometry(XRF). Then the difference between the radial striations and un-radial striations rhizomes were analyzed for their fluctuation of the inorganic elements. The results showed that there were different "key period" and "key elements" in the rhizosphere elements content of L. chuanxiong rhizome with radial and un-radial striations, and different element coordination and antagonistic relationship. The key fluctuation period of rhizosphere elements in un-radial striations group were in 0-60 and 60-150 days, of which 22 elements such as Na, Mg, Al were the key elements in 0-60 days, and 5 elements such as Sr, Hf, Pb, Co, Ce were the key elements in 60-150 days. The key fluctuation period of rhizosphere elements in radial striations group were in 0-60 and 210-270 days, of which four elements such as Na, Co, Ce, As are the key change elements in 0-60 days, and 18 elements such as Mg, Al, Si are the key change elements in 210-270 days. At the same time, the study showed that the fluctuation of inorganic elements in rhizosphere soil coincided with the growth and development process of L. chuanxiong and the key period of the formation of "radial striations rhizome". The key stage which the rapid growth of lateral buds of rhizome affected the formation of radial striations is 60-150 days after planting, while the increase of Sr and Co elements is likely to be an important reason for the expansion of lateral buds of rhizome and the failure to form typical "radial striations rhizome" in un-radial striations group.

Objectives: To investigate the prevalence and transmission of mcr-3 among Salmonella enterica serotype Typhimurium and 1,4,[5],12:i:-., Methods: A total of 4724 clinical Salmonella isolates were screened for the presence of mcr-3 in China during 2014-19. The clonal relationship of the mcr-3-positive isolates and their plasmid contents and complete sequence were also characterized based on WGS data from the Illumina and MinION platforms., Results: We identified 10 mcr-3-positive isolates, and all were MDR, mostly resistant to colistin, cefotaxime, ciprofloxacin, doxycycline and florfenicol. mcr-3 was co-present with blaCTX-M-55-qnrS1 on hybrid ST3-IncC-FII conjugatable plasmids (n = 6) and an ST3-IncC non-conjugatable plasmid (n = 1) and embedded into a pCHL5009T-like IncFII plasmid on the Salmonella chromosome (n = 3). Four distinctive genetic contexts surrounded mcr-3 and all but one were closely related to each other and to the corresponding region of IncFII plasmid pCHL5009T. IS15DI was most likely the vehicle for integration of mcr-3-carrying IncFII plasmids into ST3-IncC plasmids and the chromosome and for shaping the MDR regions. In addition, a phylogenetic tree based on the core genome revealed a unique Salmonella lineage (≤665 SNPs) that contained these 10 mcr-3-positive isolates and another 38 (33 from patients) mcr-3-positive Salmonella from five countries. In particular, most of the 51 mcr-3-positive isolates belonged to ST34 and harboured diverse antibiotic resistance genes (ARGs), including mcr-3-blaCTX-M-55-qnrS1, and possessed similar ARG profiles., Conclusions: Our findings revealed global clonal spread of MDR ST34 Salmonella from clinical isolates co-harbouring mcr-3 with blaCTX-M-55 and qnrS1 and a flexibility of mcr-3 co-transmittance with other ARGs mediated by mobile genetic elements., (© The Author(s) 2020. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Microecology was directly or indirectly involved in the growth and development, metabolism process, and component accumulation of traditional Chinese medicine(TCM) in various ways, which affected the formation and changes of the geoherbalism of TCM. It was one of the main tasks of traditional Chinese medical microecology(TCMM) to reveal the relationship among microecological structure and its change rule and the quality effect of TCM. The heterogeneity of soil environment caused by geographical and climatic factors, as well as the discreteness limitation caused by isolation factors such as distance and host selection, were the main causes of the differentiation of microecological geography of TCM. The microecology of TCM had important influences and contributions on the distinctive origin and quality of Dao-di herbs, which was mainly reflected in the formation of excellent germplasm(including disease and insect resistance, drought resistance, salt resistance, cold resistance, etc.), the increase of yield, the formation of medicinal parts, the metabolism and accumulation of effective components, the time limit of harvesting, and the toxicity, increasing efficiency or reducing toxicity of TCM in the processing, the changes of product efficiency after introduction, and the authenticity of fungus medicine. With the vigorous development of metabonomics and modern information technology, the following aspects would become the future research trends, including the microecologically mediated biogenic pathway of chemical components, the metabolic synthesis reactor of TCM based on the microecological quantitative effect relationship, the cultivation of genuine Chinese medicine based on reconstruction of microecological structure, the origin identification barcode traceability technology, and the toxicity reduction and efficiency enhancement technology of TCM based on the microecological.

Background: The aim of this study was to investigate the association of the formation of neutrophil extracellular traps (NETs) with gut leakage in type 1 (T1D) and type 2 diabetes (T2D)., Methods: In all, 105 subjects (56 T1D, 49 T2D) were included in the study. Eight biomarkers of NET formation and gut leakage (ie, protein arginine deiminase type 4 [PAD4], neutrophil elastase [NE], proteinase 3 [PR3], complement 5a [C5a], α 1 -antitrypsin [AAT], DNase I, zonulin, and lipopolysaccharide [LPS]) were measured in serum samples by ELISA. Neutrophils were isolated and stimulated by phorbol myristate acetate to form NETs in vitro. Neutrophil intracellular contents were then collected and used as antigens to detect anti-neutrophil cytoplasmic antibodies (ANCA) in the serum., Results: There was an increase in NET-associated proteins (PAD4, NE, PR3, C5a, AAT and DNase I) in new-onset T1D patients but not in those with T2D. Of PAD4, NE, and PR3, PAD4 was found to be the most sensitive biomarker for the diagnosis of T1D. Furthermore, circulating levels of zonulin and LPS were not only increased, but were also strongly correlated with NET formation and ANCA generation in T1D patients., Conclusions: This study provides evidence that increased formation of NETs, particularly PAD4, is closely associated with gut leakage in T1D but not T2D, and suggests that microorganisms and the release of neutrophil cytoplasmic antigen during the formation of NETs may be involved in the pathogenesis of T1D., (© 2018 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and John Wiley & Sons Australia, Ltd.)

It is necessary to examine the characteristics of hydrological cycle in Gaoyou area of Jiang-su Province in response to climate change and flood disasters. In this study, 121 atmospheric rain samples were collected and environmental factors were recorded from July 2015 to October 2017. We analyzed the hydrogen and oxygen stable isotopic composition of rain and identify moisture sources in this area. Results showed that the hydrogen and oxygen isotopic composition of rain had an seasonal variation, which was positive in the winter but negative in the summer. D-excess value was higher in winter than that in summer. On the annual scale, significantly negative relationships between δD (δ 18 O) and temperature and between δD (δ 18 O) and precipitation indicated the "anti-temperature effect" and "precipitation amount effect", respectively. On the seasonal scale, there was no obvious "temperature effect" but "precipitation amount effect" in autumn and winter. Results from the HYSPLIT model showed that the precipitation in this area came mainly from ocean evaporation that was influenced by the Chinese South Sea, Indian Ocean and Pacific Ocean. Precipi-tation in other seasons mainly came from the water vapor mixture from the Eurasian continent, Atlantic Ocean, Arctic Ocean, as well as local evaporation. The seasonal pattern of δD and δ 18 O values in precipitation was mainly influenced by monsoon activity and El Niño-Southern Oscillation (ENSO). In addition, the precipitation isotopes clearly indicated the shift in climates from El Niño to La Niña.

In the long-term evolution, microbes and hosts coexist widely, forming a symbiotic microecosystem and resulting the complex interactions of the metabolism. With the application of microecological theory in Chinese materia medica science, two main points have been accepted gradually. On the one hand, the prevention and treatment of human diseases by traditional Chinese medicines can be achieved through the correction and adjustment of the imbalance of the human microecosystem. On the other hand, the microecosystem can regulate the quality of traditional Chinese medicines in real time, and further affect their curative effect. Thus, a new discipline, Traditional Chinese Medical Microecology, has been gradually established. In this review, the background, theoretical structure, research directions, key problems and the relationship with human microecology of Traditional Chinese Medical Microecology were systematically summarized and prospected for promoting its development. Moreover, this review provides a reference protocol for further discoursing the microecological mechanism involving the efficacy of traditional Chinese medicines., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Background: The purpose of this article is to introduce the modified technique of preservation of the inferior alveolar nerve (IAN) during mandibulectomy for a benign lesion., Methods: Five cases of osteofibrous hyperplasia and 3 cases of centricity osteomyelitis were included. During surgery, the IAN was marked using a planned cutting guide. Using an oscillating saw, the depth of the osteotomy along the IAN was controlled until the bone cortex was cut through. After splitting, the bony section was removed, leaving the neurovascular bundle intact. The sensation of the lower lip was evaluated using current perceptive threshold testing during follow-up., Results: After follow-up for 6-27 months, no recurrence or secondary deformity was found. One patient had severe sensory disturbance., Conclusion: With the use of a cutting guide and osteotomy tricks, mandibulectomy with preservation of the IAN can be accurately performed., (© 2017 Wiley Periodicals, Inc.)

The aim of this study was to investigate the osseous characteristics of Chinese temporomandibular joint (TMJ) and detect the size clusters for total joint prostheses design.Computer tomography (CT) data from 448 Chinese adults (226 male and 222 female, aged from 20 to 83 years, mean age 39.3 years) with 896 normal TMJs were chosen from the Department of Radiology in the Shanghai 9th People's Hospital. Proplan CMF 1.4 software was used to reconstruct the skulls. Three-dimensional (3D) measurements of the TMJ fossa and condyle-ramus units with 13 parameters were performed. Size clusters for prostheses design were determined by hierarchical cluster analyses, nonhierarchical (K-means) cluster analysis, and discriminant analysis.The glenoid fossa was grouped into 3 clusters, and the condyle-ramus units were grouped into 4 clusters. Discriminant analyses were capable of correctly classifying 97.24% of the glenoid fossa and 94.98% of the condyle-ramus units. The means and standard deviations for the parameter values in each cluster were determined.Fossa depth and angles between the condyle and ramus were important parameters for Chinese TMJ prostheses design. 3D measurements and cluster analysis of the osseous morphology of the TMJ provided an anatomical reference and identified the dimensions of the minimum numbers of prosthesis sizes required for Chinese TMJ replacement.

Objective: To get an understanding of the microflora of endophytic actinomycetes in Ligusticum chuanxiong,and to obtain the resource of antagonistic strains against Ligusticum chuanxiong root rot disease., Methods: Actinomycetes in stem nodes and rhizomes of Ligusticum chuanxiong were isolated and purified by a series of means,namely tissue block method, homogenate technique and plat streaking method. Further, dual culture and inhibition zone method were employed to test the antagonistic activity of these strains. To identify strains with potential precisely,both microscopic observation and DNA sequencing were conducted., Results: 83 strains of Ligusticum chuanxiong endophytic actinomycetes were identified, which consisted of 13 species groups. Among all of them,species identified as Streptomyces scopuliridis( KF600747. 1), Streptomyces griseorubiginosus( AB706352. 1) and Streptomyces agglomeratus( LC055413. 1) showed antagonistic activities against four kinds of pathogenic fungi of Ligusticum chuanxiong root rot disease., Conclusion: Actinomycetes which belong to Griseorubroviolaceus groups of streptomyce are potential biocontrol microbes to Ligusticum chuanxiong root rot,showing importance to the production of Chuanxiong Rhizoma.

Objective: To identify the pathogens causing root rot disease on Ligusticum chuanxiong from the main producing areas in Sichuan, and to provide gist of management of this disease in the future., Methods: The diseased rhizomes of Ligusticurn chuanxiong with root rot symptom were sampled in two main producing areas in Sichuan. According to Koch's postulate, the pathogenicity on Ligusticum chuanxiong rhizomes of the isolates was confirmed. The representative isolates were identified based on their morphological characteristics and ribosomal DNA internal transcribed spacer(rDNA-ITS) sequences., Results: The isolates(f2-16, 3-2, f4-19 and f5-7)which caused root rot disease held high homology reached 99%, 99%, 100% and 100% respectively, when compared with GenBank sequences GQ229075, JQ796755, JN232136 and FJ481024., Conclusion: The causing agents of root rot disease on Ligusticum chuanxiong belong to Fusarium solani, Fusarium oxysporum, Plectosphaerella cucumerina and Phoma glomerata. Pathogens from different regions are varied.

Purpose: Piezosurgery has been used widely in oral and maxillofacial surgery, but there has been no report systematically describing an osteotomy method with piezosurgery for complicated mandibular third molar removal. The aim of this study was to introduce 4 osteotomy methods using piezosurgery and evaluate their effects., Materials and Methods: A retrospective study was conducted of patients with a complicated impacted mandibular third molar requiring extraction. The predictor variable was the extraction technique. Four osteotomy methods using piezosurgery were tested according to different impaction types: method 1 involved complete bone removal; method 2 involved segmental bone removal; method 3 involved bone removal combined with tooth splitting; and method 4 involved block bone removal. Outcome variables were success rate, operative time, major complications (including nerve injury, mandible fracture, severe hematoma, or severe edema), and serious pyogenic infection. Data were analyzed using descriptive statistics., Results: The study was composed of 55 patients with 74 complicated impacted mandibular third molars. All impacted mandibular third molars were removed successfully. The average surgical time was 15 minutes (range, 8 to 26 minutes). Thirty-eight molars (51.4%) were extracted by method 1, 18 molars (24.3%) by method 2, 12 molars (16.2%) by method 3, and 6 molars (8.1%) by method 4. Two cases (2.7%) developed postoperative infections and recovered within 1 week using drainage and antibiotic administration., Conclusion: The 4 osteotomy methods with piezosurgery provide effective ways of removing complicated impacted mandibular third molars., (Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II (BMPR2), E1A binding protein p300 (EP300), transforming growth factor-β2 (TGFβ2), and tumor necrosis factor, and alpha-induced protein 3 (TNFAIP3) gene expression patterns in B-cell malignancies were studied., Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcript-polymerase chain reaction (qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as reference., Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control (P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control (P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group (P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3 (r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control (P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines (P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells (P<0.05)., Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.

Background: Local resection of the mass was reported to treat the condylar osteochodroma in some cases. The purpose of this study was to evaluate the feasibility and the indications of the local resection., Methods: In all, 47 patients with osteochondroma of the mandibular condyle were treated from January 2002 to March 2012. The decision to perform local resection depended on 2 factors: there was a stalk existing between the mass and the condyle, and the condylar surface was involved less than 1/2., Results: Local resection of the mass was performed in 38 cases (80.1%). These masses were removed by 4 methods: direct removal (18 cases), pushed-out by a screw and steel wire (14 cases), excision in multiple pieces (3 cases), and temporary osteotomy of the zygomatic arch (3 cases). In the follow-up period, there was no recurrence., Conclusion: Local resection of the mass was a more conservative procedure to treat the solitary osteochondroma with a stalk., (Copyright © 2013 Wiley Periodicals, Inc.)

The purpose of this study was to evaluate the guiding effect of computer-assisted design technique in local resection of exostosis osteochondroma (EOC) from the mandibular condyle. Eight patients diagnosed with EOC through computed tomographic scan were selected from January 2011 to March 2012. SurgiCase CMF 5.0.0.32 software was used for preoperative design. The osteotomy angle, depth, and tumor shape were measured to guide the surgery. The tumors were resected from the stalk 2 mm within the normal tissue to restore the unaffected condyle as much as possible. Postoperative computed tomographic scans showed that the tumors had been resected completely in all patients. The mean (SD) error between the design and the result was 1.82 (1.25) mm. Five patients had more than 1-year follow-up without recurrence. Computer-assisted design is a good way to help local resection of EOC. It can improve the accuracy of tumor resection and keep the unaffected condyle as much as possible for function.

Objective: To research the diversity of endophytic fungal communities among Ligusticum chuanxiong growing at 5 areas in Sichuan province, and illuminate the developing mechanism of geoherbs from the microecological perspective., Method: The PCR-DGGE and DNA sequencing techniques were used to analyze the endophytic fungi community of L. chuanxiong., Result: The community of endophytic fungi present difference among different growing areas. Though minor difference were found among individuals at the same area, similarity among individuals from the same growing areas were higher significantly than those from different growing areas. Compared with the other 4 growing areas, L. chuanxiong from Shiyang town, Dujiangyan city had more abundant endophytic fungi and low similarity to others, and which probably had special types of fungi., Conclusion: The abundant and stable endophytic fungal community is an important factor for the development of geoherb L. chuanxiong at Shiyang town, Dujiangyan city.

Objective: To explore the functions of tumor susceptibility gene 101 (TSG101) in the invasion and metastasis of gastric cancer cells by cell culture., Methods: The TSG101 eukaryotic expression and empty plasmids were transfected into gastric cancer cell line SGC7901. After screening with G418, single cell clone was selected and cultured. The expression of TSG101 was detected by reverse transcription (RT)-PCR and Western blotting. Cells were divided into TSG101 eukaryotic expression plasmid and blank control groups. Then the relationship was examined between TSG101 expression and tumor invasion and metastasis through the invasion, mobile, adhesion and damage scar experiment., Results: The expression levels of TSG101 in mRNA and protein in the TSG101 eukaryotic expression group were significantly higher than those of the plasmid and blank control groups (0.85 ± 0.09 vs 0.55 ± 0.07, 0.45 ± 0.07 and 29.4 ± 1.2 vs 17.0 ± 0.4, 15.9 ± 0.4, all P < 0.05). The cell number of TSG101 eukaryotic expression group through Matrigel, laminin, type IV collagen protein (84 ± 14, 128 ± 10, 62 ± 7) were significantly higher than those of the plasmid group (55 ± 9, 77 ± 10, 31 ± 6) and blank control group (48 ± 8, 76 ± 9, 24 ± 5, all P < 0.01). The number of cells adherent to Matrigel, laminin, type IV collagen protein of the TSG101 eukaryotic expression group (0.97 ± 0.04, 1.34 ± 0.04, 0.90 ± 0.01) were obviously higher than those of the plasmid group (0.53 ± 0.03, 0.75 ± 0.05, 0.42 ± 0.02) and blank control group (0.60 ± 0.03, 0.72 ± 0.03, 0.40 ± 0.01, all P < 0.01). The number of TSG101 eukaryotic expression group cell migrating to membrane lower surface was obviously higher than that of the plasmid group and blank control group (87 ± 13 vs 54 ± 8, 48 ± 7, all P < 0.01). The fusion speed of the TSG101 eukaryotic expression group was faster than that of plasmid and blank control groups after cultivating for 24 and 48 h., Conclusions: TSG101 expression increases significantly in SGC-7901 cells after a stable transfection of TSG101 eukaryotic expression plasmids. Also the capacities of invasion and metastasis become markedly enhanced.

Objective: B-cell lymphoma 2 (Bcl-2) is an important member of the Bcl-2 family of proteins that regulate the induction of apoptosis. This study aims to investigate whether Bcl-2 small interfering RNA (siRNA) combined with miR-15a oligonucleotides (ODN) could enhance methotrexate (MTX)-induced apoptosis in Raji cells., Methods: Chemically synthesized miR-15a ODN and Bcl-2 siRNA were transfected in Raji cells by using a HiPerFect Transfection Reagent and then combined with MTX. Expression levels of Bcl-2 protein were detected by Western blot. Cell proliferation was determined by CCK8 assay. The rate of cell apoptosis was determined by Annexin V/PI double staining. The morphology of apoptotic cells was observed by Hoechst-33 258 staining., Results: After the cells were transfected with miR-15a ODN combined with Bcl-2 siRNA, Bcl-2 protein levels were evidently decreased. CCK8 assay showed that cell proliferation was significantly decreased and was significantly lower in miR-15a ODN combined with Bcl-2 siRNA plus MTX group than in miR-15a ODN with methotrexate group, Bcl-2 siRNA with MTX group, and single MTX group (P<0.05). Hoechst 33258 staining revealed numerous apoptotic cells. AnnexinV/PI double staining showed that the apoptotic rates were (13.13±1.60)%, (34.47±2.96)%, (32.87±3.48)%, and (45.47±2.16)% in MTX, Bcl-2 siRNA plus MTX, miR-15a ODN plus MTX, and miR-15a ODN combined with Bcl-2 siRNA plus MTX groups, respectively. Among these groups, the apoptotic rate of miR-15a ODN combined with Bcl-2 siRNA plus MTX group was the highest; this apoptotic rate was also significantly different from that of miR-15a ODN or Bcl-2 siRNA plus MTX (P<0.05)., Conclusions: Bcl-2 siRNA combined with miR-15a ODN could enhance MTX-induced apoptosis in Raji cells. Bcl-2 siRNA and miR-15a combined with MTX may be a useful approach to improve the treatment effects on lymphoma.

Objective: In order to better understand the nature of Salmonella infection in diarrheal patients in Guangdong province, the study analyzed the serum types, antibiotic resistance and molecular determinants of the isolated Salmonella strains., Methods: In year 2010, 8405 diarrhea patients from 16 surveillant hospital in Guangzhou, Zhongshan, Dongguan, Zhuhai, Maoming, Yangjiang and Jiangmen cities in Guangdong province, were recruited in the study. A total of 8405 fecal specimen were collected and subjected to Salmonella isolation and culture. The isolated Salmonella strains were further analyzed via serotyping, antimicrobial susceptibility testing, and PFGE. The χ(2) test was applied to compare the differences between the isolated Salmonella strains in different seasons and districts. BioNumerics software was used to analyze the PFGE results in order to determine the correlation between different Salmonella strains., Results: The positive rate of the surveillant Salmonella in Guangdong province was 3.58% (301/8405) in 2010; with the gender ratio at 1.34:1 (166/124). Salmonella infection was found in all age groups, and most in infants, accounting for 57.48% (173/301). The isolated rates of Salmonella were separately 3.48% (61/1751), 4.97% (134/2695), 3.08% (73/2370) and 2.08% (33/1589) in the four seasons; and the difference was statistically significant (χ(2) = 27.29, P < 0.01). The isolated rates of Salmonella in different regions were as follows: Zhuhai 15.43% (25/162), Maoming 7.53% (18/239), Dongguan 6.51% (39/599), Yangjiang 3.64% (14/385), Zhongshan 3.03% (70/2309), Guangzhou 2.90% (126/4349) and Jiangmen 2.49% (9/362). The difference between regions was statistically significant (χ(2) = 100.75, P < 0.01). Except one strain of the isolated Salmonella cannot be serotyped, the other 300 strains were divided into 42 serotypes, of which Salmonella typhimurium and Salmonella enteritidis were dominant, account for 45.18% (136/301) and 10.96% (33/301) respectively. Although over 85% of Salmonella were sensitive to cephalosporin, ACSSuT resistance patterns (defined as resistance to at least ampicillin, chloramphenicol, streptomycin, sulfamethoxazole and tetracycline) reached 34.88% (105/301), the highest resistant rate was found in serotype Salmonella typhimurium, as high as 65.44% (89/136). 136 strains of Salmonella typhimurium were divided into 51 PFGE types, showed great genetic diversity. 33 strains of Salmonella enteritidis were divided into 18 PFGE types. The strains with same PFGE pattern may have different drug-resistant patterns, and vice versa., Conclusion: Salmonella typhimurium and Salmonella enteritidis were the dominant serotypes causing infectious diarrhea in Guangdong province. Cephalosporin was the primary choice in clinical medicine. However, Salmonella typhimurium was resistant to drug most seriously in Guangdong province. There was no significant correlation between Salmonella resistance patterns and PFGE type.

Objective: To understand the distribution, molecular characteristics and virulence genes of the O1 and O139 Vibrio cholerae isolates from the Pearl River Estuary water., Methods: Vibrio cholerae isolates collected from the Pearl River estuary waters from January 2009 to December 2010, were tested by PCR for eight virulence-related genes, including cholera toxin (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), hemolysin (hlyA), toxin-coregulated pilus (tcpA), outer membrane protein (ompU), and the regulatory protein genes (tcpI, toxR). Genetic relation was assessed by pulsed-field gel electrophoresis (PFGE) and the patterns were clustered by BioNumerics., Results: From 1152 aquatic samples, 69 isolates were identified, including 41 Inaba, 18 Ogawa and 10 O139. All the isolates showed ctxA negative, while the hlyA and toxR genes were positive in all the isolates. 34.15% (14/41) of the Inaba strains were hlyA(+) toxR(+) ompU(+) ace(+) zot(+) tcpI(+), while 66.67% (12/18) belonged to Ogawa strains and 70% (7/10) of the O139 strains were hlyA(+) toxR(+). Through PFGE analysis, the O1 isolates formed three clusters in this study. The patterns of O1 isolates differed widely, with the similarity as 72.8% - 100.0%, while the patterns of O139 isolates having the similarity of 69.9% - 95.5%., Conclusion: The non-toxigenic O1 and O139 V. cholerae had a wide distribution in the environment of Pearl River estuary water during the non-epidemic period of cholera. All the aquatic isolates presented diversities on the related virulent genes.

Objective: To study the serotypes, virulence features and molecular characteristics of Vibrio parahaemolyticus isolated in food poisoning cases and surveillance program on diarrhea patients in Guangdong, 2009., Methods: 95 Vibrio parahaemolyticus strains from food poisoning cases and 15 strains from surveillance program on diarrhea patients were serotyped and detected for tdh (thermostable direct hemolysin, tdh) and trh (tdh-related hemolysin gene, trh) by PCR. 81 sero-variant Vibrio parahaemolyticus strains were selected through PFGE subtyping., Results: There were 15 Vibrio parahaemolyticus strains isolated from surveillance program on diarrhea patients and 95 strains were isolated from 11 Vibrio parahaemolyticus-caused food poisoning cases in 2009. Among these strains, O3:K6 (46.67% and 44.21%) and O4:K8 (33.33% and 28.42%) were the dominant serotypes, but not the 7 food-borne strains. There were 93 (84.54%) tdh(+)trh(-), 13 (11.81%) tdh(-)trh(-), and 3 (3.65%) tdh(+)trh(+) strains. The similarity value was between 57.7% to 100.0% of the 81 strains after PFGE sub-typing method and 36 PFGE subtypes were identified. PFGE001 and PFGE029 appeared to be the dominant subtypes., Conclusion: O3:K6 and O4:K8 were the most dominant serotypes in Vibrio parahaemolyticus-caused diarrhea and food poisoning cases in Guangdong and tdh were detected in most of the strains. Dominant PFGE subtypes were causing both sporadic and outbreak cases in different areas in Guangdong province.

Objective: To understand the phenotypic characteristics of foodborne Vibrio parahaemolyticus in Guangdong province through carrying out a comprehensive comparison including pulse field gel electrophoresis, ribotyping and serotyping., Methods: 74 different Vibrio parahaemolyticus strains isolated from seafood and cases due to food poisoning in Guangdong province were under serotyping and susceptibility testing, in addition to the testing of direct heat hemolysin (tdh) and the heat hemolysin-related hemolysin hormone (trh) via PCR. Ribosomal genotyping (ribotyping) with EcoR I restriction enzyme was utilized on 74 different Vibrio parahaemolyticus isolates, whereas pulsed-field gel electrophoresis (PFGE) with the Not I restriction enzyme was used on 74 different Vibrio parahaemolyticus isolates. BioNumerics software was used to compare the isolates from different sources, times and places in order to elicit the correlation between different strains., Results: Although Vibrio parahaemolyticus was 100.00% sensitive to chloramphenicol, it still presented different levels of resistance against 13 other antibiotics. Among the 74 different strains of Vibrio parahaemolyticus under testing, 24.32% showed positive for the tdh virulence gene, whereas 4.05% positive for trh. 74 different Vibrio parahaemolyticus strains were found to belong to 26 serotypes, where the O5:K17 and O2:K28 serotypes were dominant in those isolates that causing seafood-poisoning. The O3:K6 serotype was found to be the dominant of those isolates that causing food-poisoning. Based on ribosomal genotyping, the 74 Vibrio parahaemolyticus isolates were divided into 62 different ribotypes, whereas the 74 strains of Vibrio parahaemolyticus were divided into 67 different PFGE types, thus exhibiting considerable genetic diversities of the strains., Conclusion: Majority of the isolates causing food-poisoning carried tdh virulence gene. PFGE was shown to have the highest resolution, followed by ribotyping with serotyping being the lowest, where the combination of the three could improve the resolution.

The concurrence of 2 independent neoplasias in the ipsilateral parotid and the temporomandibular joint (TMJ) region was infrequently reported. In this article, we present a unique case characterized by the coexistence of a rare salivary gland tumor, basal cell adenoma, of the parotid gland with a ganglion cyst in the ipsilateral TMJ region. A special surgical procedure was also presented here using a modified preauricular incision and transcondylar approach for extracapsular dissection of both lesions. Previously published literature are about the traditional treatment of benign parotid tumors using partial or total parotidectomy with the preservation of facial nerve mainly via an S-shaped submandibular incision and approach. Here, for the first time, a special surgical method using preauricular transcondylar approach for this unique case of synchronous occurrence of parotid tumor and TMJ cyst is reported.

Objective: To understand the genetic polymorphism of Salmonella and Staphylococcus aureus in Guangdong province, as well as to explore methods for identifying and tracing the source of these two foodborne pathogens., Methods: Using the automated ribotyping system, two foodborne pathogens were tested with either EcoR I or Pvu II restriction enzymes. BioNumerics software was then applied for image analysis, database establishment and other corresponding analysis., Results: Digestion of 32 Salmonella isolates with Pvu II yielded 19 different ribotypes, and digestion of 14 Salmonella isolates with EcoR I yielded 2 different ribotypes. Staphylococcus aureus isolates showed greater genetic diversity, whereas EcoR I digestion of 49 different isolates yielded 31 different ribotypes., Conclusion: Unique Salmonella and Staphylococcus aureus isolates could be identified through ribotyping. Although Salmonella serotyping and ribotyping were not strongly correlated, the combination of both restriction enzymes could be used to more effectively identify the genetic relationship among different strains as well as the source of food poisoning. Thus, not only could the genetic relationships amongst the different strains be inferred through ribotyping skills, the source of food poisoning and mode of transmission could also be determined under the use of this method.

Dentigerous cysts are thought to be caused by a developmental abnormality derived from the reduced enamel epithelium of the tooth forming organ. Most typical dentigerous cysts are those associated with the third molar teeth of the mandible, but rarely involve impacted supernumerary teeth in the anterior maxilla. Swelling and/or pain may be the major complaints of the patients. Herein, we review the literature spanning the past 22 years concerning dentigerous cysts associated with supernumerary teeth in the anterior maxilla, and present four additional cases with emphasis given to the clinicopathological characteristics of this type of dentigerous cyst.

Objective: To understand the infection of Salmonella (S.) in patients with diarrhea and outbreaks caused by Salmonella to identify the serotypes, resistance to antibiotics and PFGE types of the strains from the surveillance program in Guangdong province., Methods: S. strains from patients with diarrhea were detected, and all the positive strains collected in routine and outbreak surveillance programs, were tested by serum agglutination, antibiotic susceptibility and PFGE., Results: 71 S. strains were isolated from 1922 stool samples in 2008, with positive rate as 3.7%. 85 S. strains were isolated from 2110 stool samples in 2009, with positive rate as 4.0%. All the 156 strains were divided into 37 serotypes, with S. serotype typhimurium and enteritidis as the most common serotypes. 10 incidents of food poisoning were detected, of which 4 were caused by enteritidis and 3 by typhimurium. A suspected outbreak by enteritidis was discovered and under epidemiological investigation. The findings indicated that 2 of the 4 patients from this outbreak were infected with identical enteritidis isolates. 80% of the 229 isolates were found susceptible to cephalosporins and quinolone and 59.3% of them were multiresistant to the antibiotics., Conclusion: S. enteritidis and S. typhimurium were the most common serotypes that caused infectious diarrhoea and food poisoning in Guangdong province.

Objective: To prepare a DNA Microarray that can detect 8 common species of food borne bacterial pathogens in south China., Methods: All the 70mer oligo probes were designed on the characteristic genome loci of the 8 species of food borne bacterial pathogens. Eight subarrays corresponding to the 8 food borne bacterial pathogens were spotted onto the slide and integrated into a pan-array on the chip. A number of identified and known bacterial samples from the storage bank were selected for the validation test., Results: Based on the PPR ranking, for LM sub-array, the PPR of the 3 Listeria bacteria LM, Lin and Liv was 68.8%, 51.8% and 59.6%, respectively, while that of the non-Listeria bacterial samples was all below 43%. For VC sub-array, the PPR of VC sample was 54.1% and that of the non-VC bacterial samples was lower than 17.2%. For VP sub-array, the PPR was 66.7% for VP sample and below 24.2% for non-VP bacterial samples. For Sal sub-array, the PPR was 55.9% for Sal sample and below 50.5% for non-Sal bacterial samples. For Shi sub-array, the PPR of Shi sample and the non-Shi bacterial samples was 53.8% and below 36.6%, respectively. For SA sub-array, the PPR of SA sample and non-SA bacterial samples was 65.2% and below 22.7%, respectively. For CJ sub-array, the PPR of the 2 Campylobacter bacteria CJ and CC were 88.2% and 58.8%, respectively, and that of the non-Campylobacter bacterial samples was lower than 35.3%. For EC sub-array, the PPR of EC sample was 47.9%, and that of the non-EC bacterial samples was lower than 41.6%. Evaluation of the Biosafood-8 chip developed in this study by 18 biological samples from different origins demonstrated its good specificity and accuracy in the identification of the pathogens., Conclusion: The chip we developed can clearly differentiate the target food borne pathogenic bacteria and non-target bacteria and allows specific and accurate identification of the species of the tested bacteria isolates.

Mesenchymal stem cells (MSCs) constitute an interesting cellular source to promote brain regeneration after Parkinson's disease. MSCs have significant advantages over other stem cell types, and greater potential for immediate clinical application. The aim of this study was to investigate whether MSCs from the human placenta could be induced to differentiate into dopaminergic cells. MSCs from the human placenta were isolated by digestion and density gradient fractionation, and their cell surface glycoproteins were analyzed by flow cytometry. These MSCs were cultured under conditions promoting differetiation into adipocytes and osteoblasts. Using a cocktail that includes basic fibroblast growth factor (bFGF), all trans retinoic acid (RA), ascorbic acid (AA) and 3-isobutyl-1-methylxanthine (IBMX), the MSCs were induced in vitro to become dopamine (DA) neurons. Then, the expression of the mRNA for the Nestin and tyrosine hydroxylase (TH) genes was assayed via RT-PCR. The expression of the Nestin, dopamine transporter (DAT), neuronal nuclear protein (NeuN) and TH proteins was determined via immunofluorescence. The synthesized and secreted DA was determined via ELISA. We found that MSCs from the human placenta exhibited a fibroblastoid morphology. Flow cytometric analyses showed that the MSCs were positive for CD44 and CD29, and negative for CD34, CD45, CD106 and HLA-DR. Moreover, they could be induced into adipocytes and osteocytes. When the MSCs were induced with bFGF, RA, AA and IBMX, they showed a change in morphology to that of neuronal-like cells. The induced cells expressed Nestin and TH mRNA, and the Nestin, DAT, NeuN and TH proteins, and synthesized and secreted DA. Our results suggest that MSCs from the human placenta have the ability to differentiate into dopaminergic cells.

It has been reported that genes encoding antigens of bacterial and viral pathogens can be expressed in plants and are shown to induce protection antibodies. The structural protein E2 of classical swine fever virus (CSFV), which has been shown to carry critical epitopes, has been expressed in different systems. Here, we report the expression of CFSV E2 gene in tobacco chloroplasts. Mice immunized with leaf extracts elicited specific antibodies. This indicated that the expressed E2 proteins had a certain degree of immunogenicity. To our knowledge, this is the first report showing induction of protective antibody in response to classical swine fever virus (CSFV) by immunization with antigen protein E2 expressed in tobacco chloroplasts, which will open a new way to protection from CSFV by plant chloroplasts as bioreactors.

Objective: To establish molecular typing of Listeria monocytogenes isolates by pulsed-field gel electrophoresis (PFGE) for studying the epidemiologic characteristics of Listeria monocytogenes isolated from foodstuff in Guangdong province and to build up PFGE typing database of Listeria monocytogenes isolates for identifying the infectious resource of the outbreaks and other epidemiologic investigation., Methods: "Standardized Protocol for Molecular Subtyping of Listeria monocytogenes by PFGE" was followed. BioNumerics software was applied on image analysis, database establishment, comparative and corresponding analysis., Results: 107 Listeria monocytogenes isolates were typed by PFGE, 41 PFGE types were observed among the isolates. The PFGE types were dispersive among these isolates. Listeria monocytogenes isolates were most frequently isolated in raw chicken while the most PFGE types were found in this type of food. The positive rate was relatively high in cold and iced foods. Only 1-2 DNA fragment difference occurred in 26 Listeria monocytogenes isolates by PFGE, so high degree of relatedness remained among these isolates. There were unique PFGE patterns in the regions of Shaoguan and Huizhou. From time to time, a number of isolates remained close relationship., Conclusion: PFGE typing of the 107 Guangdong Listeria monocytogenes isolates demonstrated relative genetic diversity but a number of the isolates showed close relatedness.

Bcl-2, a prominent member of the family of proteins, is responsible for dys-regulation of apoptosis and resistance to chemotherapy and radiotherapy. This study investigated whether small hairpin RNA (shRNA) targeting Bcl-2 could render A549 cells more susceptible to gamma radiation-induced apoptosis. Recombinant Bcl-2 shRNAs expression vector were transfected into A549 cells with Lipofectamine 2000. Transfected cells were screened in 800 mg/ml G418 screening medium, and after stable transfection, silencing was examined. Expression of the Bcl-2 protein was assayed using Western blot in A549 cells. Inhibition of cell growth was assessed by a MTT assay. Apoptosis was determined by morphological observation and flow cytometry. Expression levels of Bcl-2 protein from A549 cells decreased after stable transfection with Bcl-2 shRNAs. No differences in Bcl-2 protein levels between control shRNA group and untreated cells were noted. After stable transfection with Bcl-2 shRNAs the viability of cells was less than after stable transfection with those with control shRNAs and untransfected A549, respectively (P<0.05). Control shRNA had no significant effect on growth of cells. Radiation significantly inhibited the growth of cells stably transfected with Bcl-2 shRNA (P<0.05). No difference in survival between the cells with control shRNA and untransfected cells was noted. Using Giemsa staining, cells stably transfected with Bcl-2 shRNA combined with radiation at 48 h displayed changes of apoptosis. After treatment with radiation apoptotic rates of the A549 cells stably transfected with Bcl-2 shRNA significantly increased (P<0.05), compared with the cells with control shRNA and untransfected cells. shRNAs against the Bcl-2 mRNA increases radiation-induced apoptosis in A549 cells.

The expression vector, pBI121CTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed by fused PCR and transferred into potato (Solanum tuberosum L.) by Agrobacterium-mediated transformation. Transformed plants were obtained by selecting on kanamycin-resistant medium strictly and regenerated. The transgenic plantlets were identified by PCR, Southern-blot and the production of fused protein was confirmed and quantified by Western-blot and ELISA assays. The results showed that the fused genes were expressed stablely under the control of specific-tuber patatin promoter. The expressed fused proteins have a certain degree of immunogenicity.

The expression of classical swine fever virus (CSFV) structural protein E2 in different vectors, which has been shown to carry critical epitopes, has been established. Here, we reported a Chlamydomonas reinhardtii chloroplast expression vector, P64E2, containing classical swine fever virus structural protein E2 gene, which was constructed and transferred to C. reinhardtii by biolistic bombardment method. The transformants were identified by PCR, Southern blotting, Western blotting after selecting on resistant media. ELISA quantification assay showed that the expressed E2 protein accumulated up to 1.5-2% of the total soluble protein. The results of the study on the immunological activity indicated that the protein E2 expressed in C. reinhardtii chloroplasts could elicit animal bodies to produce antibodies against protein E2.

Objective: To investigate the patterns of condylar fractures associated with temporomandibular joint ankylosis (TMJA) and treatment methods and results based on the different types of ankylosis., Methods: Forty-two joints of ankylosis in 31 patients with were categorized to four groups according to Sawhney's classification and undergone surgical treatment as follows: a joint release and disc reposition for Type I ankylosis, a dissection of bony block and disc reposition for Type II; a dissection of full-joint and employment of the temporal myofascial flap as interposition for Type III; a radically dissection of full joint followed by ramus distraction osteogenesis and genioplasty for Type IV. All of patients were followed up for 9 to 54 months with an average of 30 months. The range of mouth opening and temporomandibular joint (TMJ) function were assessed. Condylar fractures were retrospectively investigated on the patterns and the course of ankylosis development. Macroscopical visualization on the osseously ankylosed sites and disc displacement were analyzed in comparison with the radiological findings., Results: Condylar sagittal and comminuted fractures were most susceptible to TMJA. Early fibrous ankylosis occurred usually at the 4th or 5th month post-traumatically with an average month opening of 18.3 mm. The articular discs were found displaced in all cases and early bony bridge formed at a limited area where there was no disc as cushion. During fellow-up, considerable improvement in mandibular movement was attained with a stable joint function and mouth opening range of over 30 mm except for two cases in which ankylosis relapsed., Conclusions: Condylar sagittal and comminuted fractures are most likely to cause ankylosis. Early surgical intervention could reduce the disc and avoid the later ankylosis.

Aim: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver cells in vitro., Methods: 14.5-day-old mouse fetal liver-derived cells were cultured, and were induced by 200 micromol/L butylated hydroxyanisole (BHA) combined with PI3K inhibitor LY294002 (20 micromol/L), and then were incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting or RT-PCR., Results: There was low level of neurofilament-L (NF-L) and brain factor-1 (BF-1) but no neurofilament-H (NF-H) and tyrosine hydroxylase (TH) in fetal liver cells. BHA promoted significantly expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver cells. NF-L mRNA increased 5.8 fold, NF-H mRNA 8.0 fold, BF-1 mRNA 2.68 fold, and TH mRNA 30 fold, respectively (all P < 0.01 vs untreated cells). NF-L protein increased 11.29 fold, NF-H 5.5 fold, BF-1 2.53 fold, TH 4.76 fold. Moreover, expression of these BHA-induced genes were inhibited by PI3K inhibitor LY294002., Conclusion: BHA induced neural differentiation of fetal liver cells through PI3K.

Background & Objective: Antisense oligodeoxynucleotides (ASODN) targeting bcl-2 have in vitro and in vivo antitumor activities. We have identified a novel antisense sequence in the coding region of bcl-2 mRNA that could enhance chemosensitivity and radiosensitivity of leukemia cells and lung carcinoma cells. This study was designed to investigate inhibitory effects of bcl-2 ASODN on development and growth ability of human lung carcinoma NCI-H460 cells xenograft in nude mice., Methods: bcl-2 ASODN was added to NCI-H460 cells. Inhibitory rate of cell growth was assayed by MTT assay. bcl-2 ASODN-treated NCI-H460 cells were implanted subcutaneously into nude mice to observe tumor growth and calculate tumor inhibitory rate. Seven days after subcutaneous implantation of untreated NCI-H460 cells, the tumor-bearing mice were randomized into 3 groups: control group, ASODN group, and nonsense oligodeoxynucleotides (NSODN) group. bcl-2 ASODN, bcl-2 NSODN, and saline were separately injected into the tumor bodies of mice in relevant groups for 3 week. The tumor weight and volume were measured, and the morphology of tumor cells was observed under microscope., Results: bcl-2 ASODN reduced the growth of NCI-H460 cells in a time-dependent manner. Inhibitory rate of NCI-H460 cells was significantly higher in bcl-2 ASODN group than in bcl-2 NSODN group (P<0.05). The tumorigenic ability of NCI-H460 cells was reduced by bcl-2 ASODN, with the maximum tumor growth inhibitory rate of 87.5% (P<0.01). The tumor formation time was prolonged to 12.6 days. Tumor growth was significantly inhibited in bcl-2 ASODN group as compared with that in NSODN group and control group(P<0.01). The tumor weight was significantly lighter in bcl-2 ASODN group than in bcl-2 NSODN group and control group (P<0.01). The growth inhibitory rate was 71.0% in bcl-2 ASODN group. Serious necrosis foci and inflammatory cell infiltration were observed in tumor tissues of bcl-2 ASODN group, while cancer cells were diffused in tumor tissues of bcl-2 NSODN group and control group., Conclusion: bcl-2 ASODN can suppress tumorigenic ability of NCI-H460 cells and inhibit tumor growth in nude mice.

Objective: To explore the clinical characteristics of naso-orbital-ethmoid (NOE) fracture, principles and surgical methods., Methods: Twenty-nine patients with NOE fractures were investigated. Operations for the treatment of NOE and subsequent deformities included reduction of central fragment, reconstruction of orbit walls, reattachment of medial canthal ligament and so on. The outcome of the treatment was evaluated., Results: Three cases were infected postoperatively. Of the 14 cases performed on reconstruction of orbit wall, 9 cases got complete correction for their dystopia/enophthalmos, 5 cases improved. Thirteen cases experiencing canthopexy obtained accepted outcome., Conclusions: The satisfactory correction of NOE fracture and subsequent enophthalmos/dystopia and telecanthus can be achieved with elective surgery in most cases.

Objective: To observe the management results of hemangioma in infants and children., Methods: Forty-nine cases of hemangioma in infants and children managed from 1986 to 2004 were reviewed. During the management period, the incidence and first visit age and sex were recorded. The location and volume of the lesions were photographed and followed up at an interval of 3 to 12 months until the complete involution of the tumor. The treatment modalities used included local injection of prednisone in 16 cases, surgery in 6 cases, and the remaining 33 cases were observed without treatment., Results: Thirty-three cases receiving no treatment involuted naturally. The complete involution age was found from 9 months to 10 years with average age of 5.4 years. The cosmetic results after involution were good., Conclusions: Most hemangiomas do not need treatment and can involute naturally. However, in cases with severe complications such as expansion, destruction and obstructive or congestive heart failure, treatment is indicated.

Aim: To analyze the correlation between the protein expression of p16 and Rb genes in gastric carcinoma (GC), to investigate the role of p16 gene in invasion and lymph node metastasis of GC, and to examine the deletion and mutation in exon 2 of p16 gene in GC., Methods: The protein expression of p16 and Rb genes was examined by streptavidin-peroxidase conjugated method (S-P) in normal gastric mucosa, dysplastic gastric mucosa and GC. The deletion and mutation of p16 gene were examined by polymerase chain reaction (PCR) and polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) respectively in normal gastric mucosa and GC., Results: The positive rates of P16 and Rb protein expression respectively were 96% (77/80) and 99% (79/80) in normal gastric mucosa, 92% (45/50) and 80% (40/50) in dysplastic gastric mucosa, 48% (58/122) and 60% (73/122) in GC. The positive rates of P16 and Rb protein expression in GC were significantly lower than that in normal gastric mucosa and dysplastic gastric mucosa (P<0.05). The positive rate of P16 protein expression in mucoid carcinoma (10%, 1/10) was significantly lower than that in poorly differentiated carcinoma (51%, 21/41), undifferentiated carcinoma (58%, 15/26) and signet ring cell carcinoma (62%, 10/16) (P<0.05). The positive rates of P16 protein in 30 cases of paired primary and lymph node metastatic GC were 47% (14/30) and 17% (5/30) respectively, being significantly lower in the later than in the former (P<0.05). There was no mutation in exon 2 of p16 gene in the 25 freshly resected primary GCs. But five cases in the 25 freshly resected primary GCs displayed deletion in exon 2 of p16 gene. The positive rate of both P16 and Rb proteins was 16% (14/90), and the negative rate of both P16 and Rb proteins was 8% (7/90) in 90 GCs. The rate of positive P16 protein with negative Rb protein was 33% (30/90). The rate of negative P16 protein with positive Rb protein was 43% (39/90). There was reverse correlation between P16 and Rb expression in 90 GCs (P<0.05)., Conclusion: The loss protein expression of p16 and Rb genes is related to GC. The loss expression of P16 protein is related to the histopathologic subtypes and lymph node metastasis of GC. Expression of P16 and Rb proteins in GC is reversely correlated. The deletion but not mutation in exon 2 of p16 gene may be involved in GC.

Objectives: The value of supraomohyoid neck dissection used in treating oral squamous cell carcinoma was discussed., Methods: Twenty-seven cN0 patients with oral squamous cell carcinoma were entered into the study. Supraomohyoid neck dissection was performed to remove the lymphatic tissue of level I, II, and III. The operation duration and shoulder function were recorded., Results: The average operation duration was (1.6 +/- 0.2) h. Nineteen percent (5/27) of the cN0 neck were proved positive pathologically which included two cases in level I and four in level II (one case had both level I and II metastases). Shoulder function recovered in three months after operation. All patients were followed up from two years to four years and none of them had local or neck recurrence., Conclusions: Supraomohyoid neck dissection is a right choice for cN0 patients with oral squamous cell carcinoma with its advantages in both curing neck lymphatic metastases and preserving neck and shoulder contours and functions.

Objective: To investigate a new classification and treatment of zygomatic complex fractures (ZCF) by clinical and radiographic studies., Methods: Standard radiographs with Waters, submental vertex views had been obtained before and after operation in 206 patients (212 sides) with ZCF. Patients with complicated fractures underwent two and three dimensional CT. Computer-assisted measuring system was used to measure the degree of displacement., Results: ZCF were classified into 3 types and 6 subtypes according to the degree of displacement of segments and facial deformities. Treatment principles were proposed. The results were satisfied through clinical application and measurement of post-operative images., Conclusions: The treatment for ZCF should be selected according to the fracture types. The key of operation is to recover the malar prominence. Remodeling the length and radian of zygomatic arch guarantee the recovery of malar prominence. Zygomatic-sphenoid and zygomatic-maxillary fissures are very important reference marks for reduction and fixation.

To explore the effect of antisense phosphorothioate oligodeoxynucleotide (ASODN) of human telomerase reverse transcriptase (hTERT) gene on telomerase activity in CEM cells, PCR enzyme-linked immunoassay was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by RT-PCR and immunofluorescence assay using fluoresce isothiocyanate label respectively. The results showed that the expression levels of hTERT mRNA and protein in CEM cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and sense oligodeoxynucleotide-treated cells. Telomerase activity decreased when CEM cells were treated with ASODN for 48 hours. Telomerase activity of CEM cells was significantly inhibited when treated with ASODN for 72 hours. There was no difference in telomerase activity levels between control and hTERT sense oligodeoxynucleotide-treated cells. These results suggested that hTERT ASODN inhibited telomerase activity of CEM cells.

Background & Objective: The previous study has identified two novel antisense oligonucleotides (AS-ODN) of new target point in the translation initiation and the coding region of bcl-2 mRNA that could increase the sensitivity of HL-60 and K562 cells lines to etoposide, daunorubicin, and araninosyl cytosine (Ara-C). This study was designed to investigate whether the two novel bcl-2 AS-ODNs could increase Ara-C-induced apoptosis of cultured primary acute leukemia (AL) and chronic lymphocytic leukemia (CLL) Cells., Methods: Cell surviving fraction was determined using the trypan blue dye exclusion assay. The expression levels of bcl-2 protein were assayed by immunofluorescence using fluorescein isothiocyanate label. Apoptosis was detected by Giemsa staining and flow cytometry., Results: Treatment with the two AS-ODNs/Ara-C combination respectively for 48 h significantly inhibited the growth of AL and CLL cells. There was significant difference on AL and CLL cells survival between nonsense oligodeoxynucleotide(NS-ODN)/Ara-C combination and Ara-C treated cells alone (P < 0.05). It was found that the two AS-ODNs respectively combined with Ara-C could significantly downregulate bcl-2 protein expression (P < 0.05). There was a significant increase in apoptotic rates of AL or CLL cells after treatment with the two AS-ODNs respectively combined with Ara-C as respectively compared with the NS-ODN/Ara-C combination and Ara-C treated cells alone; and the effect had statistical difference (P < 0.05). Compared to the AS-ODN directed against the translation initiation of bcl-2 mRNA, the AS-ODN directed against the coding region showed stronger effect in the aspects of increasing the sensitivity of AL cells to Ara-C (P < 0.05)., Conclusions: The two novel AS-ODNs of new target point in the translation initiation and the coding region of bcl-2 mRNA could enhance Ara-C induced apoptosis of AL and CLL cells.

To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on telomerase activity in primary acute myeloid leukemia (AML) cells and chronic myeloid leukemia (CML) cells, polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) was used to determine telomerase activity. The expression levels of hTERT protein were assayed by flow cytometry using immunofluorescence labeling. Immunofluorescence assay showed that the expression levels of hTERT protein in AML and CML cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT protein levels between control and sense oligodeoxynucleotide (SODN)-treated cells. Telomerase activity decreased when AML and CML cells were treated with ASODN for 48 h. Telomerase activity of AML and CML cells was significantly inhibited when the cells treated with ASODN for 72 h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells. It was concluded that hTERT ASODN could inhibit hTERT protein expression level, thus decreasing the telomerase activity in primary AML and CML cells.

Background & Objective: Human telomerase reverse transcriptase (hTERT) is a limiting component for telomerase activity. Telomerase inhibitor targeting telomerase RNA could arrest growth of tumor cells. This study was designed to explore the effect of hTERT gene antisense phosphorothioate oligodeoxynucleotide (ASODN) on telomerase activity in HL-60 cells., Methods: Telomeric repeat amplification protocol was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence using fluorescence isothiocyanate label respectively., Results: RT-PCR and immunofluorescence assay showed that the expression levels of hTERT mRNA and protein from HL-60 cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and SODN-treated cells. Telomerase activity decreased when HL-60 cells were treated with ASODN for 48 h. Telomerase activity of HL-60 cells was significantly inhibited when treated with ASODN for 72h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells., Conclusion: Telomerase activity could be down-regulated by hTERT ASODN in HL-60 cells.