Search

Background: Lymph node status is a prominent prognostic factor for intrahepatic cholangiocarcinoma (ICC). However, the prognostic value of performing lymph node dissection (LND) in patients with clinical node-negative ICC remains controversial. The aim of this study was to evaluate the clinical value of LND on long-term outcomes in this subgroup of patients., Methods: We retrospectively analyzed patients who underwent radical liver resection for clinically node-negative ICC from three tertiary hepatobiliary centers. The propensity score matching analysis at 1:1 ratio based on clinicopathological data was conducted between patients with and without LND. Recurrence-free survival (RFS) and overall survival (OS) were compared in the matched cohort., Results: Among 303 patients who underwent radical liver resection for ICC, 48 patients with clinically positive nodes were excluded, and a total of 159 clinically node-negative ICC patients were finally eligible for the study, with 102 in the LND group and 57 in the non-LND group. After propensity score matching, two well-balanced groups of 51 patients each were analyzed. No significant difference of median RFS (12.0 vs. 10.0 months, P = 0.37) and median OS (22.0 vs. 26.0 months, P = 0.47) was observed between the LND and non-LND group. Also, LND was not identified as one of the independent risks for survival. Among 51 patients who received LND, 11 patients were with positive lymph nodes (lymph node metastasis (LNM) (+)) and presented significantly worse outcomes than those with LND (-). On the other hand, postoperative adjuvant therapy was the independent risk factor for both RFS (hazard ratio (HR): 0.623, 95% confidence interval (CI): 0.393 - 0.987, P = 0.044) and OS (HR: 0.585, 95% CI: 0.359 - 0.952, P = 0.031). Furthermore, postoperative adjuvant therapy was associated with prolonged survivals of non-LND patients (P = 0.02 for RFS and P = 0.03 for OS)., Conclusions: Based on the data, we found that LND did not significantly improve the prognosis of patients with clinically node-negative ICC. Postoperative adjuvant therapy was associated with prolonged survival of ICC patients, especially in non-LND individuals., Competing Interests: The authors declare that they have no conflict of interest., (Copyright 2024, Sha et al.)

Rapid accumulation of repair factors at DNA double-strand breaks (DSBs) is essential for DSB repair. Several factors involved in DSB repair have been found undergoing liquid-liquid phase separation (LLPS) at DSB sites to facilitate DNA repair. RNF168, a RING-type E3 ubiquitin ligase, catalyzes H2A.X ubiquitination for recruiting DNA repair factors. Yet, whether RNF168 undergoes LLPS at DSB sites remains unclear. Here, we identified K63-linked polyubiquitin-triggered RNF168 condensation which further promoted RNF168-mediated DSB repair. RNF168 formed liquid-like condensates upon irradiation in the nucleus while purified RNF168 protein also condensed in vitro. An intrinsically disordered region containing amino acids 460-550 was identified as the essential domain for RNF168 condensation. Interestingly, LLPS of RNF168 was significantly enhanced by K63-linked polyubiquitin chains, and LLPS largely enhanced the RNF168-mediated H2A.X ubiquitination, suggesting a positive feedback loop to facilitate RNF168 rapid accumulation and its catalytic activity. Functionally, LLPS deficiency of RNF168 resulted in delayed recruitment of 53BP1 and BRCA1 and subsequent impairment in DSB repair. Taken together, our finding demonstrates the pivotal effect of LLPS in RNF168-mediated DSB repair., Competing Interests: Competing interests statement:The authors declare no competing interest.

Twelve new austalide meroterpenoids ( 1 - 12 ) were isolated from the endophytic fungus Diaporthe sp. XC1211. Their structures were elucidated by extensive spectroscopic analysis. The absolute configurations of compounds 1 , 3 , 4 , and 6 were established by single-crystal X-ray diffraction, whereas those for the others were established by experimental electronic circular dichroism (ECD) data analysis. Compounds 1 - 12 represent a rare class of austalides with a 24α-CH 3 . Compounds 2 and 5 demonstrated potent proliferation inhibitory effects against LPS-induced B cells with IC 50 values of 6.7 (SI = 3.6) and 3.8 (SI > 13) μM, respectively. Compounds 2 and 5 decreased the secretion of IL-6 in LPS-induced B cells in a dose-dependent manner.

Quaternary ammonium compounds (QACs) are one type of widely used cationic biocide, and their usage amount is growing rapidly due to the flu and COVID-19 pandemic. Many QACs were released into the environment in or after the course of their use, and thus they were widely detected in water, sediment, soil, and other environmental media. QACs have stronger surface activity and non-specific biotoxicity, which poses a potential threat to the ecosystem. In this study, the environmental fate and potential toxicity of QACs were documented in terms of their migration and transformation process, biological toxicity effects, and the main mechanisms of bacterial resistance to QACs. Aerobic biodegradation was the main natural way of eliminating QACs in the environment, and the reaction was mainly initiated by the hydroxylation of C atoms at different positions of QACs and finally mineralized to CO 2 and H 2 O through decarboxylation, demethylation, and β-oxidation reaction. Toxicological studies showed that QACs at environmental concentrations could not pose acute toxicity to the selected biotas but threatened the growth and reproduction of aquatic organisms like Daphnia magna . Their toxicity effects depended on their molecular structure, the tested species, and the exposed durations. Additionally, our team first investigated the toxicity effects and mechanisms of QACs toward Microcystis aeruginosa , which showed that QACs depressed the algae growth through the denaturation of photosynthetic organelles, suppression of electron transport, and then induction of cell membrane damage. In the environment, the concentrations of QACs were always lower than their bactericidal concentrations, and their degradation could induce the formation of a concentration gradient, which facilitated microbes resistant to QACs. The known resistance mechanisms of bacteria to QACs mainly included the change in cell membrane structure and composition, formation of biofilm, overexpression of the efflux pump gene, and acquisition of resistance genes. Due to the similar targets and mechanisms, QACs could also induce the occurrence of antibiotic resistance, mainly through co-resistance and cross-resistance. Based on the existing data, future research should emphasize the toxicity effect and the potential QACs resistance mechanism of microorganisms in real environmental conditions.

The rapid recognition of DNA double-strand breaks (DSBs) by the MRE11/RAD50/NBS1 (MRN) complex is critical for the initiation of DNA damage response and DSB end resection. Here, we show that MRN complex interacting protein (MRNIP) forms liquid-like condensates to promote homologous recombination-mediated DSB repair. The intrinsically disordered region is essential for MRNIP condensate formation. Mechanically, the MRN complex is compartmentalized and concentrated into MRNIP condensates in the nucleus. After DSB formation, MRNIP condensates move to the damaged DNA rapidly to accelerate the binding of DSB by the concentrated MRN complex, therefore inducing the autophosphorylation of ATM and subsequent activation of DNA damage response signaling. Meanwhile, MRNIP condensates-enhanced MRN complex loading further promotes DSB end resection. In addition, data from xenograft models and clinical samples confirm a correlation between MRNIP and radioresistance. Together, these results reveal an important role of MRNIP phase separation in DSB response and the MRN complex-mediated DSB end resection., (© 2022. The Author(s).)

Introduction: Increasing evidence has indicated that LINC00680 represents an oncogenic factor in cancer; however, the mechanism by which LINC00680 contributes to breast cancer (BC) remains unknown. Methods: A dual-luciferase reporter assay was used to explore the relationship between LINC00680, miR-320b, and cyclin-dependent kinase 5 (CDKL5). A CCK-8 assay and transwell assay were utilized to evaluate the proliferation and invasion in docetaxel-resistant BC cells, respectively. Results: LINC00680 and CDKL5 protein levels were both upregulated when induced by different concentrations of docetaxel. LINC00680 knockdown decreased the expression level of drug resistance-related genes, proliferation, and invasion of BC cells. Bioinformatics prediction and dual-luciferase assays revealed that miR-320b targeted the 3 ' -unstranslated regions (UTR) of both LINC00680 and CDKL5, suggesting that the modulation of LINC00680 on CDKL5 occurred via sequestering miR-320b. Conclusion: Overall, this study highlights the important role of LINC00680 in docetaxel resistance through the miR-320b/CDKL5 pathway and provides a novel therapeutic strategy for BC drug resistance.

Objective: To evaluate the regulation efficacy of oral tangeretin on testosterone and cortisol in sprinters at winter training season. Methods: Twenty-four sprinters were paired and randomly divided into experimental group (EG) and control group (CG). During winter training season, EG were treated with 200 mg tangeretin by oral, and CG were treated with placebo for 4 weeks. Blood samples were collected on the first day of each week (T1, T2, T3, T4) and after the intervention (T5) to detect serum levels of testosterone, cortisol, superoxide dismutase (SOD), and adrenocorticotropic hormone (ACTH). The body composition was tested at T1 and T5. Results: After 4 weeks, ①the serum cortisol level of CG was increased, and the serum levels of testosterone and SOD were decreased significantly ( P ＜0.05). ②In EG, the serum levels of cortisoland ACTH were decreased significantly ( P ＜0.05, P ＜ 0.01), while the serum testosterone level was remained stable, and the level of SOD was increased slightly. ③The muscle mass of EG and CG were increases, but that of EG was increased higher than that of CG. Conclusion: Tangeretin reduces the oxidative stress response that caused by high-intensity exercise during winter training, which maintain the serum testosterone level and inhibit cortisol excessive secretion and promote muscle synthesis.

Objective: To introduce our experience of laparoscopic uterovaginal anastomosis and operative outcomes following this technique and update the clinical management of congenital cervicovaginal atresia., Study Design: Between March 2015 and January 2019, twenty-three patients with congenital cervicovaginal atresia underwent laparoscopic uterovaginal anastomosis and Luohu procedure. Their clinical characteristics, surgical outcomes and follow-up data were retrospectively reviewed., Results: The mean ± SD age of the cases was 16.4 ± 4.0 (11-26) years. All patients had a history of primary amenorrhea and cyclical abdominal pain, and the average delay in diagnosis from first symptoms was 33.5 ± 38 (1-156) months. Most of the patients had 1-3 cm long vaginal pouch. The operative procedure lasted 125 ± 32 (80-190) min. The average vaginal length at 1 month was 7.9 ± 1.3 (range 6-9) cm. All patients showed resumption of menstruation. The patients were followed for a mean of 27 ± 13 (12-56) months. During the follow-up, cervical stenosis did not occur in any of the cases., Conclusion: Laparoscopic uterovaginal anastomosis with Luohu procedure provided a minimally invasive, safe, and effective surgical option for the patients with congenital cervicovaginal atresia. The technique is uncomplicated, easy to learn and perform, and provides a functional and anatomic satisfactory result. No special surgical instruments are required with this technique., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The regulation of mTOR and the dimethylation of histone H3 on lysine 9 (H3K9me2) H3K9me2 by Uhrf1 and the mechanism of autophagy regulation in myocardial ischemia-reperfusion injury (MIRI) were studied in vivo and in vitro . An in vitro I/R injury model was established using the primary mouse cardiomyocytes treated with H 2 O 2 . Subsequent analysis by qRT-PCR, western blot, and immunofluorescence indicated that overexpression of Uhrf1 significantly inhibited apoptosis of the H 2 O 2 -treated cardiomyocytes, reduced expression of apoptosis factors caspase-3 and Bax, and increased expression of apoptosis inhibitory factor Bcl-2. Furthermore, Uhrf1 was found to increase cardiomyocyte proliferation and promote the expression of mTOR, while the four expression peaks of H3K9me2 on the mTOR gene were inhibited by overexpression of Uhrf1. The expression of autophagy factors LC3, Beclin-1, and p-mTOR in Uhrf1-overexpressed cardiomyocytes was dramatically increased, and P62 expression was dramatically decreased. When an H3K9me2 inhibitor was added to the Uhrf1-knockdown cardiomyocytes, the expression of mTOR was increased, the expression of LC3, Beclin-1, and p-mTOR was decreased, and P62 expression was significantly increased. In the present study, Uhrf1 exhibits a protective function in MIRI, reducing the apoptosis of cardiomyocytes while increasing their proliferation and viability.

Arginine methylation is an important posttranslational modification catalyzed by protein arginine methyltransferases (PRMTs). However, the role of PRMTs in colorectal cancer (CRC) progression is not well understood. Here we report that non-POU domain-containing octamer-binding protein (NONO) is overexpressed in CRC tissue and is a potential marker for poor prognosis in CRC patients. NONO silencing resulted in decreased proliferation, migration, and invasion of CRC cells, whereas overexpression had the opposite effect. In a xenograft model, tumors derived from NONO-deficient CRC cells were smaller than those derived from wild-type (WT) cells, and PRMT1 inhibition blocked CRC xenograft progression. A mass spectrometry analysis indicated that NONO is a substrate of PRMT1. R251 of NONO was asymmetrically dimethylated by PRMT1 in vitro and in vivo. Compared to NONO WT cells, NONO R251K mutant-expressing CRC cells showed reduced proliferation, migration, and invasion, and PRMT1 knockdown or pharmacological inhibition abrogated the malignant phenotype associated with NONO asymmetric dimethylation in both KRAS WT and mutant CRC cells. Compared to adjacent normal tissue, PRMT1 was highly expressed in the CRC zone in clinical specimens, which was correlated with poor overall survival in patients with locally advanced CRC. These results demonstrate that PRMT1-mediated methylation of NONO at R251 promotes CRC growth and metastasis, and suggest that PRMT1 inhibition may be an effective therapeutic strategy for CRC treatment regardless of KRAS mutation status.

To evaluate the ability of inulin to enhance the immune response of a ptfA gene DNA vaccine for avian Pasteurella multocida, inulin was added as an adjuvant to the ptfA-DNA vaccine, obtaining an inulin-adjuvant DNA vaccine. The DNA vaccine was administered to chickens; a fimbria protein vaccine and an attenuated live vaccine were used as positive controls. The levels of the serum antibody and concentrations of interferon-γ (IFN-γ), interleukin-2 (IL-2), and interleukin-4 (IL-4) were determined, and a lymphocyte proliferation assay was performed. After being challenged with virulent P. multocida, the protective efficacy was evaluated. The results showed that the serum antibodies induced by the ptfA-DNA vaccine were not enhanced by inulin. The stimulation index values and the concentrations of IL-2 and IFN-γ in chickens vaccinated with inulin-adjuvant DNA vaccine were significantly higher than those in chickens vaccinated with the DNA vaccine, those with the fimbria protein vaccine, and the chickens gavaged with inulin. The concentrations of IL-4 in the inulin-adjuvant DNA vaccine group and the fimbria protein vaccine group were higher than those in the DNA vaccine group and the inulin-gavage group. The protective efficacy rates of the attenuated live vaccine group, the fimbria protein vaccine group, the DNA vaccine group, the inulin-adjuvant DNA vaccine group, and the inulin-gavage group were 90, 70, 55, 65, and 55%, respectively., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Scrophularia ningpoensis has been used as a famous traditional medicinal herb in Asian countries to treat jaundice, dysentery, and the pain of rheumatism. In this paper, the complete chloroplast (cp) genome sequence of S. ningpoensis was reported and characterized. The cp genome is 153,175 bp in length, composed of a pair of 25,490 bp inverted repeat (IR) regions separated by a large single copy (LSC) region of 84,257 bp and a small single copy (SSC) region of 17,938 bp. There were 130 predicted genes (85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes) in the genome, and the overall GC content of the genome is 38%. Phylogenetic analysis based on the cp genome data showed that S. ningpoensis was sister to S. buergeriana., Competing Interests: No potential conflict of interest was reported by the authors, (© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Background: Hepatocellular carcinoma (HCC) is one of the main causes of cancer-related death. This study aims to explore the role and underlying mechanism of H19 in HCC., Methods: qRT-PCR detected miR-15b-5p and H19 expression, as well as the mRNA level of EMT-associated genes. Western blotting detected protein level of EMT-associated genes. Immunohistochemistry (IHC) examined CDC42 in HCC tissues. Dual luciferase reporter assay verified the regulatory mechanism among H19, miR-15b and CDC42. Colony formation, wound healing assay, transwell, flow cytometry measured proliferation, migration, invasion and apoptosis, respectively., Results: H19 and CDC42 were up-regulated while miR-15b was down-regulated in HCC cells and tissues. miR-15b interacted with H19 and CDC42 3'-UTR. H19 knockdown inhibited proliferation, migration and invasion, and increased apoptosis, which was rescued by miR-15b inhibitor. H19 knockdown suppressed CDC42/PAK1 pathway and EMT progress., Conclusion: H19 knockdown inhibited proliferation, migration and invasion, and promoted apoptosis of HCC cells via targeting miR-15b/CDC42/PAK1 axis., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: The aim of this study was to use whole genome sequencing (WGS) help detect de novo mutations or pathogenic genes of Mayer-Rokitansky-Küster-Hauser syndrome type 1(MRKH syndrome type 1)., Study Design: This was a case-parent trios study. Nine unrelated probands, with MRKH syndrome type 1 and their parents were enrolled. The enrollment, sequencing process, establishment of the de novo mutations detecting procedure and experiment part were performed over a 2-year period., Results: we detected 632 de novo single nucleotide variants (SNVs), 267 de novo small insertions/deletions (indels), 39 de novo structural variations (SVs) and 28 de novo copy number alterations (CNAs). Three novel damaging coding de novo SNVs with three damaging coding de novo genes (PIK3CD, SLC4A10 and TNK2) were revealed. Two SNVs were annotated of the promoter region of gene NBPF10 and 3'UTR of NOTCH2NL, potentially contributing to the pathogenesis of MRKH., Conclusion: We identified five de novo mutations in BAZ2B, KLHL18, PIK3CD, SLC4A10 and TNK2 by performing WGS, the functional involvement of all deleterious mutations in MRKH candidate genes of the trios warrant further study. WGS may complement conventional array to capture the complete landscape of the genome in MRKH., Competing Interests: A signed declaration of no conflict of interest has been provided by all authors involved in the study.

Pseudomonas aeruginosa (P. aeruginosa) is a zoonotic pathogen that can infect a variety of animals, including poultry. However, as there is no commercial vaccine available it is imperative that new and effective vaccines are developed. In this study, 2 monovalent DNA vaccines (pOPRL and pOPRF), one divalent combination DNA vaccine (pOPRL+pOPRF) and one fusion DNA vaccine (pOPRLF) were constructed based on the oprL and oprF genes of P. aeruginosa. These vaccines were administered to chickens, an outer membrane protein vaccine (OMP vaccine) and inactivated vaccine used as positive controls. The serum antibody, interferon-γ (IFN-γ), interleukin-2 (IL-2) and interleukin-4 (IL-4) concentrations were determined and lymphocyte proliferation assays were performed. After challenging with virulent P. aeruginosa, protective efficacy was evaluated. Following vaccination, serum antibodies, stimulation index (SI) values, concentrations of IL-2 and IFN-γ in chickens vaccinated with the bivalent combination DNA vaccine and fusion DNA vaccine were found to be significantly higher than in those chickens vaccinated with the 2 monovalent DNA vaccines. Moreover, the immune indexes in the bivalent combination DNA vaccine group were higher than those in the fusion DNA vaccine group. However, the concentrations of IL-4 in the 4 DNA vaccine groups were of no significant difference. The protective efficacy rate provided by pOPRL, pOPRF, pOPRLF, pOPRL+pOPRF, inactivated vaccine and OMP vaccine were 53.3%, 40%, 66.7%, 80%, 93.3%, and 80%, respectively. The results indicate that DNA vaccines constructed with the oprL and oprF genes of P. aeruginosa, particularly the divalent combination DNA vaccine, represent better potential vaccines. This study has laid a foundation for the design and application of future DNA vaccines of P. aeruginosa.

Background: Locally advanced lung cancers involving both right upper lung lobe and superior vena cava (SVC) requiring both lung resection and SVC reconstruction are generally deemed unresectable. However, previous evidence has proved that such patients could benefit from surgery if radical resection is achieved. Generally, a hemi-clamshell approach is adopted to complete such resection. However, it has the limitation of insufficient exposure of posterior mediastinum. Therefore, we introduced a two-step surgical approach combining sternotomy and thoracotomy for such lung cancers., Methods: A two-step surgical approach combining median sternotomy and subsequent posterolateral thoracotomy, via which radical lobectomy with systematic lymphadenectomy and SVC reconstruction could be successfully achieved, was described., Results: We have performed such surgery via the two-step surgical approach combining median sternotomy and subsequent posterolateral thoracotomy in five patients from January 2017 to March 2018. All those patients achieved radical resection of the lung cancer with lobectomy and systematic lymphadenectomy and SVC reconstruction with artificial blood vessels, and had an uneventful postoperative recovery without any major complications., Conclusions: Our initial experience proved that this two-step surgical approach combining median sternotomy and subsequent posterolateral thoracotomy was safe and feasible for locally advanced lung cancers requiring both lung resection and SVC reconstruction., Competing Interests: Conflicts of Interest: This study has been presented as oral presentation at the 26th Annual Meeting of the Asian Society for Cardiovascular and Thoracic Surgery (May 24–27, 2018, Moscow, Russia).

Background: Whether lobe-specific lymph node dissection (L-SLND) could serve as an alternative to systematic lymph node dissection (SLND) in treating early-stage non-small cell lung cancer (NSCLC) remains unclear. Therefore, we conducted this comprehensive meta-analysis to compare the effect of L-SLND with that of SLND in treating early-stage NSCLC., Methods: A systematic literature search in PubMed and Embase was conducted to identify relevant studies up to 30 November 2017. Data including 5-year overall survival (OS) and disease-free survival (DFS) rates, recurrence rates, and morbidity rate were extracted and analysed., Results: A total of six studies [one randomized controlled trial (RCT) and five retrospective cohort studies] consisting of 2,037 patients with early-stage NSCLC were included for analysis. Meta-analysis showed that there was no significant difference of 5-year OS [81.7% and 79.5%, respectively; risk ratio (RR) =1.021; 95% confidence interval (CI), 0.977-1.068; P=0.352] and DFS (76.4% and 69.9%, respectively; RR =1.061; 95% CI, 0.999-1.128; P=0.054) between patients treated with L-SLND and those with SLND. Moreover, there was also no significant difference of total recurrence rates (24.3% and 25.8%, respectively; RR =0.892; 95% CI, 0.759-1.048; P=0.166) and loco-regional recurrence rates (7.9% and 9.3%, respectively; RR =0.851; 95% CI, 0.623-1.162; P=0.310) between patients treated with L-SLND and those with SLND. However, patients treated with L-SLND yielded a significant lower morbidity rate than those treated with SLND (10.2% and 13.5%, respectively; RR =0.681; 95% CI, =0.521-0.888; P=0.005)., Conclusions: L-SLND yielded a significantly lower risk of morbidity compared to SLND without compromising long-term oncologic outcomes based on available studies with relatively poor quality. L-SLND may serve as an alternative to SLND in treating early-stage NSCLC. Further well-conducted RCTs, however, are badly needed to confirm and update our conclusions., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.

A best evidence topic in thoracic surgery was written according to a structured protocol. The question addressed was 'Does high body mass index (BMI) have any impact on survival of patients undergoing oesophagectomy for oesophageal cancer?' A total of 232 papers were found using the reported search, of which 8 papers represented the best evidence to answer the clinical question, which included 1 meta-analysis and 7 cohort studies. The authors, journal, date and country of publication, patient group studied, study type, relevant outcomes and results of these papers are tabulated. One meta-analysis and 4 cohort studies provided the evidence that high BMI was significantly correlated with a better survival of oesophageal cancer patients undergoing oesophagectomy, while the other 3 cohort studies found that high BMI had no impact on the survival of those patients. We conclude that patients with high BMI may have a better prognosis than those with normal BMI undergoing oesophagectomy for oesophageal cancer.

Background: Hepatectomy and additional common bile duct exploration are required for the treatment of left-sided hepatolithiasis (LSH)., Methods: Eligible LSH patients (n = 62) scheduled for open left lateral segmentectomy or left hemihepatectomy with intraoperative biliary exploration via the left hepatic duct orifice (LHD group, n = 35) or the common bile duct (CBD group, n = 27) were retrospectively studied. T-tube insertion was performed on selected patients. Primary outcome measures included overall operative time, length of hospital stay, intraoperative complications, residual stones, and postoperative bile leaks., Results: There were no residual stones observed in the 2 groups. Ten patients in the CBD group received T-tube placement, whereas no patients in the LHD group received T-tube placement. There were more patients in the CBD group suffered intraoperative complications and postoperative bile leakage than LHD group (P < .05). The LHD group had a significantly shorter operative time and hospitalization than the CBD group (P < .05)., Conclusion: For left-sided hepatolithiasis patients with a history of biliary tract surgery, LHD cholangioscopy is an accessible technique that simplifies the operation procedure by avoiding choledochotomy and subsequent T-tube insertion, which results in lower complication rates as well as shorter operative duration and length of hospitalization., (Copyright © 2017 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved.)

The aim of this study was to evaluate the effect of chitosanon the immune response induced by a DNA vaccine based on the ptfA gene of avian Pasteurella multocida. Naked DNA vaccine was packed with chitosanmolecules, resulting in a chitosannanoparticle DNA vaccine. The encapsulation efficiency, shape, size and resistance to DNA degradation were determined. The vaccine was administered to chickens and serum antibody, interferon-γ (IFN-γ), interleukin-2 (IL-2) and interleukin-4 (IL-4) concentrations were determined and lymphocyte proliferation assays were performed. After challenge with virulent avian P. multocida, protective efficacy was evaluated. The encapsulation efficiency of the chitosan nanoparticle DNA vaccine was 95.3%. The particle size was approximately 200nm and close to spherical in shape and it could effectively resist degradation by DNases. Following vaccination, serum antibodies, stimulation index (SI) value and concentrations of IFN-γ and IL-2 in chickens vaccinated with the chitosan nanoparticle DNA vaccine were significantly higher than those that were vaccinated with the naked DNA vaccine (P-values are 0.026, 0.045, 0.039 and 0.024, respectively). However, the concentrations of IL-4 in the two DNA vaccines group were no significant difference (P=0.157). The protective efficacy rate provided by naked DNA vaccine, chitosan nanoparticle DNA vaccine and the attenuated live vaccine were 56%, 68% and 88%, respectively. The results indicated that chitosan was able to enhance the immune response to a naked DNA vaccine based on the ptfA gene of P. multocida., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Enamelin (ENAM) has three putative phosphoserines (pSers) phosphorylated by a Golgi-associated secretory pathway kinase (FAM20C) based on their distinctive Ser-x-Glu (S-x-E) motifs. Fam20C-knockout mice show severe enamel defects similar to those in the Enam-knockout mice, implying an important role of the pSers in ENAM. To determine the role of pSer 55 in ENAM, we characterized ENAM Rgsc514 mice, in which Ser 55 cannot be phosphorylated by FAM20C due to an E 57 >G 57 mutation in the S-x-E motif. The enamel microstructure of 4-week-old mice was examined by scanning electron microscopy. The teeth of 6-day-old mice were characterized by histology and immunohistochemistry. The protein lysates of the first lower molars of 4-day-old mice were analyzed by Western immunoblotting using antibodies against ENAM, ameloblastin and amelogenin. ENAM Rgsc514 heterozygotes showed a disorganized enamel microstructure, while the homozygotes had no enamel on the dentin surface. The N-terminal fragments of ENAM in the heterozygotes were detained in the ameloblasts and localized in the mineralization front of enamel matrix, while those in the WT mice were secreted out of ameloblasts and distributed evenly in the outer 1/2 of enamel matrix. Surprisingly, the ~15  kDa C-terminal fragments of ameloblastin were not detected in the molar lysates of the homozygotes. These results suggest that the phosphorylation of Ser 55 may be an essential posttranslational modification of ENAM and is required for the interaction with other enamel matrix molecules such as ameloblastin in mediating the structural organization of enamel matrix and protein-mineral interactions during enamel formation.International Journal of Oral Science (2017) 9;e4; doi:10.1038/ijos.2017.41; published online 29 November 2017.

Background  Choledocholithiasis can be managed by transcystic (TC) and transduct (TD) stone extraction or using cholangioscopy through the left hepatic duct orifice (LHD). Objective  The aim of this study is to evaluate the safety and effectiveness of common bile duct exploration through the TC approach, TD approach, and LHD approach for choledocholithiasis, with a specific emphasis on the TC and LHD approaches versus the TD approach. Methods  Between January 2011 and June 2014, a total of 172 choledocholithiasis patients accompanied by cholecystitis and/or left intrahepatic gallstones were scheduled for laparoscopic or open common bile duct (CBD) exploration using cholangioscopy through the CBD (TD group: n = 72), cystic duct (TC group: n = 63), or LHD orifice (LHD group: n = 37). T-tube insertion was performed in selected patients. Patients were regularly followed up at bimonthly intervals or more frequently in presence of any symptom. Primary outcomes measures included overall operative time, length of hospital stay, and postoperative bile leaks. Results  Successful bile duct clearance was 100 % in the TD group, 93.6 % in the TC group, and 90.9 % in the LHD group. Sixteen cases in the TD group had T-tube placement in contrast to no cases in the TC and LHD groups. There were more bile leaks after TD stone extraction (12.5 %) than TC (3.2 %) and LHD stone extraction (0 %), which prolonged hospitalization in the TD group more than in the TC and LHD groups. For choledocholithiasis patients accompanied by cholecystitis, 2 groups (TC and TD groups) were comparable in operative time. However, for choledocholithiasis patients accompanied by left intrahepatic gallstones, the LHD group had a significantly shorter operative time than the TD group (121.1 ± 16.9 minutes vs. 149.3 ± 42.8 minutes, p < 0.05). Conclusion  The TD group had a higher stone clearance rate but was associated with a higher risk of bile leaks. TC and LHD stone extraction, which seems to be the more effective approach with lower complication rates, is an accessible technique that simplifies the operation procedure by avoiding choledochotomy and subsequent T-tube insertion., Competing Interests: Disclosure The authors report no conflicts of interest in this work., (© Georg Thieme Verlag KG Stuttgart · New York.)

Background: Choledocholithiasis can be managed by endoscopic retrograde cholangiopancreaticography/endoscopic sphincterotomy (ERCP/EST) or laparoscopic common bile duct (CBD) exploration by transcystic (TC) or transductal (TD) stone extraction., Objective: The aim of this study was to evaluate the safety and effectiveness of common bile duct stones extraction by ERCP/EST, TC approach and TD approach for choledocholithiasis, with specific emphasis on ERCP/EST, TC approach versus TD approach., Methods: Between January 2011 and June 2014, a total of 161 patients were scheduled for two-stage (preoperative ERCP/EST followed by cholecystectomy, ERCP group, n = 52)or single-stage (laparoscopic exploration of the CBD combined with cholecystectomy, n = 109) treatment for choledocholithiasis with concomitant cholecystitis. Laparoscopic common bile duct exploration was performed by TC approach (TC group, n = 63)or TD approach (TD group, n = 46). T-tube insertion was performed in selected patients. Patients were regularly followed up at bimonthly intervals or more frequently in presence of any symptom. Primary outcomes measures included length of hospital stay, successful bile duct clearance, postoperative/procedural morbidity and mortality., Results: Successful bile duct clearance was 100.0% in TD group, 93.7% in TC group and 92.3% in ERCP group. 4 cases in the TC group and 4 cases in the ERCP group required an extra choledocholithotomy due to impacted stones. 9 patients underwent T-tube drainage in TD group comparing to 1 case in ERCP group and no cases in TC group. Comparing to TC group, there was more postoperative morbidity in TD and ERCP group. Bile leaks were more frequent in TD group (8.7%) than TC (3.2%) and ERCP group (3.8%), which prolonged hospitalization in TD group than TC and ERCP group. 2 patients in ERCP group suffered duodenal perforation and one of them died because of the complication. However, total procedural morbidity was 0% in TC and TD group., Conclusion: TD stone extraction has a higher stone clearance but with a higher risk of bile leaks. Procedural morbidity is more often happened in ERCP/EST, which may result in serious consequences. TC stone extraction, which seems an effective approach with lower complication rates, is accessible techniques simplifying the operation procedure by avoiding choledocholithotomy and subsequent T-tube insertion., (Copyright © 2017 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

Unlabelled: Mussels attach to various submerged surfaces by using the byssus, which contains different proteins and is a promising source of water-resistant bio-adhesives for potential use in biotechnological and medical applications. The protein composition of the byssus has not yet been fully understood although at least eleven byssal proteins were characterized previously. In order to increase genomic resources and identify new byssal proteins from mussel Mytilus coruscus, high-throughput Illumina sequencing was undertaken on the foot, and 79,997,776 paired-ends reads were generated, yielding a library containing 88,825ft unigenes. The M. coruscus byssus was divided into three parts, the proximal thread, the distal thread, and the plaque. Byssal proteins from each part of the byssus were analyzed by shotgun-LTQ analysis. The MS/MS spectra were searched against the foot unigenes dataset and 48 byssal proteins were identified from the M. coruscus byssus. From the whole set, 17, 5, and 11 proteins were exclusive to the proximal thread, the distal thread, and the plaque, respectively. These data can be used as a resource for further studies on the roles of byssal proteins in the deposition of different byssus parts (thread vs. plaque) or in the different mechanical properties (tenacity vs. adhesion)., Biological Significance: Byssal proteins are the major component that controls different aspects of the byssal formation process and thus a source of bioactive molecules that would offer interesting perspectives in biomaterials and bio-adhesive fields. In this paper, we characterized the protein set from different partsof Mytilus coruscus byssus by a combination of transcriptome/proteome technical. A whole set of 48 byssal proteins were described here, including proteins of collagen-like, C1q domain-containing, protease inhibitor-like, tyrosinase-like, SOD, and others. Thread (the distal portion and the proximal portion) and plaque showed distinct protein composition. Of the whole byssal protein set, 11 are exclusive to the plaque, 17 are exclusive to the proximal thread, and 5 are exclusive to the distal thread. Only four proteins are shared by all the three parts of the byssus. The new byssal proteins reported here represent a significant expansion of the knowledge base of Mytilus byssal proteins, and are important for further exploring the mechanism of adhesion in mussel., (Copyright © 2016 Elsevier B.V. All rights reserved.)

FAM20A has been studied to a very limited extent. Mutations in human FAM20A cause amelogenesis imperfecta, gingival fibromatosis and kidney problems. It would be desirable to systemically analyse the expression of FAM20A in dental tissues and to assess the pathological changes when this molecule is specifically nullified in individual tissues. Recently, we generated mice with a Fam20A-floxed allele containing the beta-galactosidase reporter gene. We analysed FAM20A expression in dental tissues using X-Gal staining, immunohistochemistry and in situ hybridization, which showed that the ameloblasts in the mouse mandibular first molar began to express FAM20A at 1 day after birth, and the reduced enamel epithelium in erupting molars expressed a significant level of FAM20A. By breeding K14-Cre mice with Fam20A(flox/flox) mice, we created K14-Cre;Fam20A(flox/flox) (conditional knock out, cKO) mice, in which Fam20A was inactivated in the epithelium. We analysed the dental tissues of cKO mice using X-ray radiography, histology and immunohistochemistry. The molar enamel matrix in cKO mice was much thinner than normal and was often separated from the dentinoenamel junction. The Fam20A-deficient ameloblasts were non-polarized and disorganized and were detached from the enamel matrix. The enamel abnormality in cKO mice was consistent with the diagnosis of amelogenesis imperfecta. The levels of enamelin and matrix metalloproteinase 20 were lower in the ameloblasts and enamel of cKO mice than the normal mice. The cKO mice had remarkable delays in the eruption of molars and hyperplasia of the gingival epithelium. The findings emphasize the essential roles of FAM20A in the development of dental and oral tissues.

Avian Pasteurella multocida is the causative agent of fowl cholera, a disease much affecting the poultry industry. In order to study the efficacy of the recombinant subunit vaccine constructed with ptf A gene of avian P. multocida , the ptf A gene fragment amplified by PCR from avian P. multocida was cloned into the prokaryotic expression vector pET32a and the recombinant plasmid pET32a-ptfA was obtained. The pET32a-ptfA was expressed in Escherichia coli BL21(DE3) and the target protein rPtfA was purified. The purified protein was then mixed with Freund's adjuvant and the recombinant subunit vaccine was obtained. Three groups of chickens labeled as rPtfA, attenuated live vaccine and PBS were vaccinated with the recombinant subunit vaccine, attenuated live vaccine and PBS, respectively. Serum antibodies, peripheral blood lymphocyte proliferation (PBLP) and interferon-γ (IFN-γ) level secreted by peripheral blood lymphocyte were tested. The immunized chickens were finally challenged with virulent avian P. multocida and the protection rate was counted. Indirect ELISA showed the levels of antibodies in rPtfA and attenuated vaccine groups were most significantly higher than the other groups (P<0.01), and the former was slightly lower than the latter. Peripheral blood lymphocyte proliferation experiments and IFN-γ experiments indicated that SI value and the levels of IFN-γ induced by ConA in the two vaccine groups were significantly higher than those of the PBS groups (P<0.01), and that the attenuated vaccine group was higher than the rPtfA group. The protection rates of rPtfA and attenuated live vaccines were 45% and 75%, respectively. The results indicated that the PtfA recombinant subunit vaccine was capable of improving the immunity level and inducing a protective effect for the vaccinated chickens, but it was barely satisfactory.

Our previous studies have demonstrated that Fam20C promotes differentiation and mineralization of odontoblasts, ameloblasts, osteoblasts and osteocytes during tooth and bone development. Ablation of the Fam20C gene inhibits bone and tooth growth by increasing fibroblast growth factor 23 in serum and causing hypophosphatemia in conditional knockout mice. However, control and regulation of the expression of Fam20C are still unknown. In this study, we generated a transgenic reporter model which expresses green fluorescence protein (GFP) driven by the Fam20C promoter. Recombineering was used to insert a 16 kb fragment of the mouse Fam20C gene (containing the 15 kb promoter and 1.1 kb of exon 1) into a pBluescript SK vector with the topaz variant of GFP and a bovine growth hormone polyadenylation sequence. GFP expression was subsequently evaluated by histomorphometry on cryosections from E14 to adult mice. Fluorescence was evident in the bone and teeth as early as E17.5. The GFP signal was maintained stably in odontoblasts and osteoblasts until 4 weeks after birth. The expression of GFP was significantly reduced in teeth, alveolar bone and muscle by 8 weeks of age. We also observed colocalization of the GFP signal with the Fam20C antibody in postnatal 1- and 7-day-old animals. Successful generation of Fam20C-GFP transgenic mice will provide a unique model for studying Fam20C gene expression and the biological function of this gene during odontogenesis and osteogenesis.

Mytilus is an economically important bivalve and its shell is a biomineralized tissue with various microstructures/layers. In the present study, the shell of marine mussel, Mytilus coruscus, was analyzed and three shell layers with different morphologies and polymorphs were observed, which includes nacre, fibrous prism, and myostracum strongly attached by adductor muscles to the interior of the shell surface. In order to understand whether these different shell layers contain different shell matrix proteins (SMPs), the transcriptome sequencing of M. coruscus mantle and a parallel proteomic analysis of SMPs in the three shell layers were performed. A combination of LC-MS/MS analysis with the mantle transcriptome dataset search resulted in the identification of a total of 63 proteins from M. coruscus shell. From this protein set, fifteen, fourteen, and eight proteins were found to be unique to nacre, fibrous prism, and myostracum layers, respectively. In addition, many novel shell proteins were also identified. The data in this study could be used as a background to explore the roles of SMPs in the deposition of different shell layers (nacre vs. fibrous prism vs. myostracum), the different polymorphisms of calcium carbonate (aragonite vs. calcite); and further, the identified proteins from the myostracum could provide candidates for studying the mechanism of adductor muscle-shell attachment., Biological Significance: In this paper, we characterized for the first time the protein set from different shell layers in Mytilus. Shell matrix proteins are the major component that controls different aspects of the shell formation process and thus a source of bioactive molecules that would offer interesting perspectives in biomaterials and biomedical fields. Our data can be used as a resource for further exploring the roles of shell matrix proteins in the deposition of different shell layers (nacre vs. fibrous prism vs. myostracum) or different polymorphisms of calcium carbonate (aragonite vs. calcite), and the identified protein set of myostracum provided candidates for studying the mechanism of adductor muscle-shell attachment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Using reverse phase high performance liquid chromatography (RP-HPLC), a novel antimicrobial peptide with 55 amino acid residues was isolated from the hemolymph of Mytilus coruscus. This new antimicrobial peptide displays predominant antimicrobial activity against fungi and Gram-positive bacteria. The molecular mass and the N-terminal sequence of this peptide were analyzed by Mass Spectrometry and Edman degradation, respectively. This antimicrobial peptide, with molecular mass of 6621.55 Da, is characterized by a chitin-biding domain and by 6 Cysteine residues engaged in three intra-molecular disulfide bridges. The full-length of cDNA sequence of this new peptide was obtained by rapid amplification of cDNA ends (RACE) and the encoded precursor was turn out to be a chitotriosidase-like protein. Therefore, we named the precursor with mytichitin-1 and the new antimicrobial peptide (designated as mytichitin-CB) is the carboxyl-terminal part of mytichitin-1. The mRNA transcripts of mytichitin-1 are mainly detected in gonad and the expression level of mytichitin-1 in gonad was up-regulated and reached the highest level at 12 h after bacterial challenge, which was 9-fold increase compared to that of the control group. These results indicated that mytichitin-1 was involved in the host immune response against bacterial infection and might contribute to the clearance of invading bacteria., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Deletion or mutation of dentin matrix protein 1 (DMP1) leads to hypophosphatemic rickets and defects within the dentin. However, it is largely unknown if this pathological change is a direct role of DMP1 or an indirect role of phosphate (Pi) or both. It has also been previously shown that Klotho-deficient mice, which displayed a high Pi level due to a failure of Pi excretion, causes mild defects in the dentinal structure. This study was to address the distinct roles of DMP1 and Pi homeostasis in cell differentiation, apoptosis and mineralization of dentin and enamel. Our working hypothesis was that a stable Pi homeostasis is critical for postnatal tooth formation, and that DMP1 has an antiapoptotic role in both amelogenesis and dentinogenesis. To test this hypothesis, Dmp1-null (Dmp1(-/-)), Klotho-deficient (kl/kl), Dmp1/Klotho-double-deficient (Dmp1(-/-)/kl/kl) and wild-type (WT) mice were killed at the age of 6 weeks. Combinations of X-ray, microcomputed tomography (μCT), scanning electron microscopy (SEM), histology, apoptosis and immunohistochemical methods were used for characterization of dentin, enamel and pulp structures in these mutant mice. Our results showed that Dmp1(-/-) (a low Pi level) or kl/kl (a high Pi level) mice displayed mild dentin defects such as thin dentin and a reduction of dentin tubules. Neither deficient mouse line exhibited any apparent changes in enamel or pulp structure. However, the double-deficient mice (a high Pi level) displayed severe defects in dentin and enamel structures, including loss of dentinal tubules and enamel prisms, as well as unexpected ectopic ossification within the pulp root canal. TUNEL assay showed a sharp increase in apoptotic cells in ameloblasts and odontoblasts. Based on the above findings, we conclude that DMP1 has a protective role for odontoblasts and ameloblasts in a pro-apoptotic environment (a high Pi level).

Objective: To investigate the technique and therapeutic effect of laparoscopy-insisted vaginoplasty with peritoneum in patients with androgen insensitivity syndrome., Methods: From May. in the Fifth People' s Hospital of Shenzhen. The therapeutic effect was retrospectively analyzed., Results: Laparoscopy-insisted vaginoplasty was successfully completely with peritoneum in patients with androgen in 4 cases. Ileumtivity segyndroment was used instead of peritoneum in one case. Open operation was not adopted in any cases. The ectopic testicles were removed during operation in 4 cases. The average operation time and bleeding volume was 60 min and 20 ml, respectively. Rectum, bladder and urethra were not injured in any case. The average vaginal length was 9 cm (range 8-10 cm) 21-28 days after operation. 6 months after operation, the surface of reconstructed vagina was smooth, ruddy and flexible, with satisfactory anatomical and functional results. Normal sexual activity was achieved in 2 cases., Conclusions: Laparoscopy-insisted vaginoplasty with peritoneum could be used for female patients with androgen insensitivity syndrome. The ectopic testicles should be removed. Estrogen supplement and psychological guide after operation are also important.

Owing to their low elastic moduli, high specific strength and excellent processing characteristics in the undercooled liquid state, metallic glasses are promising materials for applications in micromechanical systems. With miniaturization of metallic mechanical components down to the micrometer scale, the importance of a native oxide layer on a glass surface is increasing. In this work we use TEM and XPS to characterize the structure and properties of the native oxide layer grown on Ni(62)Nb(38) metallic glass and their evolution after annealing in air. The thickness of the oxide layer almost doubled after annealing. In both cases the oxide layer is amorphous and consists predominantly of Nb oxide. We investigate the friction behavior at low loads and in ambient conditions (i.e. at T = 295 K and 60% air humidity) of both as-cast and annealed samples by friction force microscopy. After annealing the friction coefficient is found to have significantly increased. We attribute this effect to the increase of the mechanical stability of the oxide layer upon annealing.

Objective: To determine the accuracy and clinical value of combining 64 multi-slice spiral computer tomography (MSCT) and serum amyloid A protein (SAA) in the preoperative staging of rectal cancer., Methods: Prospectively enrolled patients with rectal cancer from October 2007 to October 2008. The patients were randomly assigned into two groups: MSCT and SAA combined group: both MSCT and SAA combinative assessment were performed for preoperative evaluation; MSCT group: only MSCT was performed preoperatively for tumor staging. The accuracy of the preoperative T, N, M, and TNM staging and the concordance rate of predictive operative strategy were compared between the two groups., Results: Total of 225 cases with rectal cancer were enrolled in this study. There were 110 cases in MSCT and SAA combined group and 115 cases in MSCT group. The baseline characteristics was comparable between the two groups. For MSCT and SAA combined group, the accuracies of preoperative staging of T, N, M and TNM was 87.3%, 85.2%, 100% and 86.4%, respectively; and for MSCT group, the corresponding rates was 85.2%, 67.0%, 100% and 66.1%, respectively. Statistical differences was found in the accuracy of preoperative N and TNM staging between the two groups (P = 0.009 and 0.001, respectively). In addition, there was statistical difference in the accuracy of prediction to operative procedures between the two groups (94.7% vs. 81.7%, P = 0.003)., Conclusion: Combinative assessment of MSCT and SAA could improve the accuracy of preoperative staging, and thus provide higher predictive coincidence rate of operative procedures.

Background: A20, also known as tumor necrosis factor alpha induced protein 3 (TNFaip3), is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B (NF-kappaB) activity and prevents tumor necrosis factor (TNF)-mediated programmed cell death. NF-kappaB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-kappaB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma (ACC) of the salivary glands., Methods: The ability of overexpression of A20 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction (PCR) and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium (MTT) and NF-kappaB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber., Results: pEGPFN3-A20 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFalpha, the NF-kappaB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells (P < 0.05). A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group (P < 0.05). The ACC-2-A20 cells showed significantly reduced ability to invade through Matrigei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% (P < 0.05)., Conclusions: A20 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-kappaB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.

Objective: To investigate the prevalence and clinical significance of hepatitis C virus (HCV) infection in patients with systemic lupus erythematosus (SLE)., Methods: Serodiagnosis was conducted in 134 SLE patients and 200 volunteer blood donors to examine the antibodies of HCV with enzyme-linked immunosorbent assay-3 (ELISA-3). Recombinant immunoblot assay-3 (RIBA-3) and PCR were performed to verify the results., Results: HCV infection was present in 15 patients with SLE (11.8%) and in 2 volunteer donors (1%, P<0.001). Compared with the SLE patients without HCV infection, the patients with HCV infection had a lower rate of cutaneous SLE features and dsDNA positivity (P=0.01 and P<0.001), but with higher incidences of hepatic damage (P<0.001) and low levels of C4 and CH50 (P=0.01 and P=0.03) as well as cryoglobulins levels (P=0.03)., Conclusion: The prevalence of HCV infection is higher in SLE patients than in non-SLE subjects, and SLE patients with positive HCV show a lower rate to present cutaneous SLE features and positive dsDNA antibody, but who have higher possibilities of hepatic damage, hypocomplementemia and cryglobulinemia.

Objective: To observe the effect of the crude extracts of Dan-shen root and San-qi of different proportion on platelet aggregation and adhesion in normal rabbits., Method: With rabbits, ig. (4d, exsanguinated via carotid artery, percentage of platelet aggregation and adhesion was measured., Result and Conclusion: Aspirin (4.4 mg/Kg) could markedly inhibit platelet aggregation and adhesion in normal rabbit. The proportions of Dan-shen root/San-qi (10:0, 10:1, 10:3, 10:6, 1:10) could markedly inhibit platelet aggregation, among which 10:3 was the best. San-qi alone had little effect on aggregation. The proportions of Dan-shen root/San-qi (10:3, 10:6, 0:10) could markedly inhibit platelet adhesion, among which 0:10 was the best, and the proportions(10:0, 10:1, 1:10) had little effect.

The Apicomplexan parasite Toxoplasma gondii replicates by endodyogeny, an unusual form of binary fission. We tested the role of TPK2, a homologue of the CDC2 cyclin-dependent kinases, in cell cycle regulation. TPK2 tagged with HA epitope (TPK2-HA-wt) was expressed in mammalian cells as confirmed by Western blot analysis using HA tag and PSTAIRE antibodies. TPK2-HA-wt phosphorylated a peptide from Histone H1, proving that TPK2 is a functional kinase. TPK2-HA-wt coimmunoprecipitated with mammalian cyclins A, B1, D3 and E. Despite being a functional kinase, TPK2 did not rescue Schizosaccharomyces pombe cdc2 and Saccharomyces cerevisiae cdc28 mutant strains. Overexpression of a dominant-negative mutant of TPK2 (TPK2-HA-dn) in T. gondii tachyzoites arrested replication. FACS analysis of tachyzoites expressing TPK2-HA-dn revealed an increase in the fraction of cells in S-phase when compared with TPK2-HA-wt transfected parasites. Expression of TPK2-HA-wt did not arrest tachyzoite replication. No discernable G2 cell cycle block was evident suggesting that cell cycle checkpoints differ in T. gondii from most other eukaryotic cells. These data suggest that TPK2 executes an essential function in T. gondii cell cycle and is likely to be the T. gondii CDC2 orthologue.

A mixture of heparin-Sepharose-purified bovine bone morphogenetic protein and type I atellocollagen was implanted in the subcutaneous tissues of 4-week, 10-month and 18-month-old rats. The implants were removed at 7, 14 and 21 days after implantation. The effects of rat age on ectopic bone formation were evaluated on the explants using haematoxylin-eosin staining, morphometric analysis, alkaline phosphatase activity and calcium content determination, as well as immunohistochemical staining of type IV collagen present in the basement membrane of blood vessels. On day 14 and 21, bone was observed in 4-week and 10-month-old rats, but the amount of bone formed in the latter was less than in the 4-week-old rats. In 18-month-old rats, bone was first found focally in very limited regions of the explants on day 21 and the amount of bone was much less than in 4-week-old rats. At all periods, alkaline phosphatase activity was higher in younger rats. On day 7, there were more blood vessels in the explants of 4-week-old rats than in those of 10-month or 18-month-old rats. On day 14 and 21, more blood vessels were found in the central regions of the explants in 4-week-old rats than in the same regions in 10-month or 18-month-old rats. The findings in the present study indicate that the rate and quantity of ectopic bone formation are reduced in aged rats, and suggest that the difference in blood vessel distribution is related to this reduction in ectopic bone formation.

As an initial effort to dissect the signaling pathways responsible for pathogenesis of Toxoplasma gondii infection, we report the cloning and in vitro functional studies of TPK3 (Toxoplasma protein kinase-3), a homologue of shaggy/glycogen synthase kinase-3 (GSK-3) kinases. The shaggy/GSK-3 family of kinases are highly conserved protein kinases that play important roles in cell fate determination, nuclear signaling and hormonal regulation. The TPK3 gene was isolated by RT-PCR with degenerate primers corresponding to highly conserved regions of serine/threonine protein kinases. The complete sequences of genomic and cDNA clones indicated the open reading frame, 1185 bp in size, is interrupted by five introns. The predicted protein sequence of TPK3 shows 54% identity to shaggy/GSK-3 over the catalytic domains. Southern analysis revealed TPK3 is a single copy locus in the Toxoplasma genome. Antisera to other GSK-3 proteins from other species recognized GST-TPK3 and a protein of the predicted size in parasites lysates. In vitro kinase assays with GST-TPK3 indicated that TPK3 autophosphorylates and phosphorylates protein phosphatase inhibitor-2 (I-2), a specific substrate of GSK-3 kinase.

We have cloned a novel human brain inward rectifier K+ channel (hIRK1), which shares approximately 60% amino acid identity with another human inward rectifier (hIRK2) but 98% identity with the mouse IRK1. The hIRK1 mRNA is expressed in several human tissues: skeletal muscle > placenta > heart > brain > lung > kidney. In human brain, the hIRK1 mRNA is uniformly distributed (except for a higher level in the corpus callosum, which contains white matter and glial cells), whereas the hIRK2 mRNA is expressed in major regions of the basal ganglia and limbic system. Xenopus oocytes injected with hIRK1 cRNA expressed an inwardly rectifying K+ current that was blocked by extracellular Ba2+. The hIRK1 channel carried a significant outward current when membrane potential was more positive than the K+ equilibrium potential (EK) and therefore had an "N-shape" current-voltage relation, resembling that of the native cardiac IRK channel. The resting membrane potential was near EK in oocytes expressing hIRK1, but was approximately -40 mV in H2O-injected or non-injected oocytes. The ability of hIRK1 to set the resting membrane potential depended on the outward current. Single-channel conductance of hIRK1 was 32 pS measured with 150 mM KCl in the patch pipette, significantly higher than 23 pS measured for mouse IRK1 and approximately 10 pS for hIRK2.

The results showed that Huanglion decoction has protective effect on ethanol-,HCl- and aspirin-induced gastric hemorrhagic lesions in rats and antemetic effect on CuSO4-induced vomiting in pigeons. A dose of 27g/(kg.d) po applied in mice showed no toxic action. This dose is 400 times that of clinical application.

Pharmacological studies were conducted on Uncaria sinensis and Achyranthes bidentata both separately and combined. Comparison was made on the hypotensive effect on normal and renal-type hypertensive rats as well anti-spasmodic and sedative effects in mice. The results showed that Uncaria sinensis and Achyranthes bidentata have obvious synergic action in compatibility.

A total of 14 cases of clear cell carcinoma of salivary glands were evaluated by immunohistochemical methods using monoclonal antibodies to cytokeratin (K1.1 and K8.12), vimentin, S-100 alpha and beta subunits, neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), MAM-3 and MAM-6 antigens and proliferating cell nuclear antigen (PCNA), as well as polyclonal antibodies to lysozyme (Ly), lactoferrin (la) and Alpha-1-antichymotrypsin (alpha 1-Ach). Histopathologically, the carcinoma was characterized by round or polygonal tumor cells with cytoplasm that does not stain with hematoxylin and eosin, nuclei with little pleomorphism and few or no mitotic figures, and growing in solid sheets, small nests or cords with collagenous stroma. Cytokeratin KL1 and K8.12 was present in few tumor cells with almost negligible to strong reaction in all cases, vimentin in 6, GFAP in 5 cases with multiple-expression of cytokeratin K8.12, vimentin and GFAP in 5 cases. S-100 protein immunoreactivity was the most prominent feature with more intense reaction of S-100 beta than S-100 alpha subunit. NSE reactivity was seen in 6 cases. Ly, La, a1-ch, MAM-3 and MAM-6 antigens were localized in clear cells with various reaction intensities. The authors conclude that the clear tumor cells in clear cell carcinoma of salivary glands are not myoepithelial in origin but epithelial or neuroectodermal/neural crest in origin, showing ductal differentiation at the immunohistochemical level.