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Infection poses a significant barrier to effective wound repair, leading to increased inflammatory responses that ultimately result in incomplete and prolonged wound healing. To address this challenge, numerous antibacterial ingredients have been incorporated into dressings to inhibit wound infection. Our previous work demonstrated that lysozyme/silver nanoparticles (LYZ/AgNPs) complexes, prepared using an eco-friendly one-step aqueous method, exhibited excellent antibacterial efficacy with favorable biosafety. To further explore its potential application in advancing wound healing, calcium alginate (CA) with good porosity, water absorption, and water retention capacities was formulated with LYZ/AgNPs to prepare composite sponge (CA/LYZ/AgNPs). As expected, in vivo experiments involving full-thickness skin wound and scald wound healing experiments demonstrated that CA-LYZ-AgNPs composite sponges with excellent biocompatibility exhibited remarkable antibacterial activity against gram-positive bacteria, gram-negative bacteria and fungi, and outperformed the wound healing process efficacy of other commercially available AgNPs-loaded wound dressings. In summary, this work introduces a CA/LYZ/AgNPs sponge featuring exceptional antibacterial efficacy and biocompatibility, thus holding promising potential in wound care applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Objective: Comparisons between patellar eversion (PE) and lateral retraction (LR) in total knee arthroplasty (TKA) are still inconclusive. To determine the most suitable procedure, we aimed to evaluate the safety and efficacy of PE and LR in TKA in this meta-analysis., Materials and Methods: This meta-analysis complied with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Web-based literature databases, including WANFANG, VIP, CNKI, the Cochrane Library, Embase, and PubMed, were utilized to conduct a comprehensive literature search for studies published until June 2022 that compared PE with LR in primary TKA. The quality of the selected randomized controlled trials (RCTs) was evaluated using guidelines of the Cochrane Reviews Handbook 5.0.2., Results: A total of 10 RCTs, including 782 patients and 823 TKAs, were selected in this meta-analysis. Our results showed that using LR improved postoperative knee extensor function and range of motion (ROM). In addition, PE and LR resulted in similar clinical benefits in terms of Knee Society Function score, pain, length of hospital stay, Insall-Salvati ratio, the occurrence of patella baja, and complications related to the operation., Conclusions: Existing evidence suggested that using LR in TKA improved early postoperative knee function. Similar clinical and radiographic outcomes were obtained 1 year after the procedures were performed. Based on these findings, we recommended the use of LR in TKA. However, studies with large sample sizes are needed to validate these findings.

The abnormal accumulation of fused in sarcoma (FUS) is a pathological hallmark in a proportion of patients with frontotemporal dementia and amyotrophic lateral sclerosis. Therefore, the clearance of FUS aggregates is a possible therapeutic strategy for FUS-associated neurodegenerative diseases. This study reports that curcumin can strongly suppress FUS droplet formation and stress granule aggregation of FUS. Fluorescence spectra and isothermal titration calorimetry showed that curcumin can bind FUS through hydrophobic interactions, thereby reducing the β-sheet content of FUS. Aggregated FUS sequesters pyruvate kinase, leading to reduced ATP levels. However, results from a metabolomics study revealed that curcumin changed the metabolism pattern and differentially expressed metabolites were enriched in glycolysis. Curcumin attenuated FUS aggregation-mediated sequestration of pyruvate kinase and restored cellular metabolism, consequently increasing ATP levels. These results indicate that curcumin is a potent inhibitor of FUS liquid-liquid phase separation and provide novel insights into the effect of curcumin in ameliorating abnormal metabolism.

The biological effects of magnetic fields (MFs) have been a controversial issue. Fortunately, in recent years, there has been increasing evidence that MFs do affect biological systems. However, the physical mechanism remains unclear. Here, we show that MFs (16 T) reduce apoptosis in cell lines by inhibiting liquid-liquid phase separation (LLPS) of Tau-441, suggesting that the MF effect on LLPS may be one of the mechanisms for understanding the "mysterious" magnetobiological effects. The LLPS of Tau-441 occurred in the cytoplasm after induction with arsenite. The phase-separated droplets of Tau-441 recruited hexokinase (HK), resulting in a decrease in the amount of free HK in the cytoplasm. In cells, HK and Bax compete to bind to the voltage-dependent anion channel (VDAC I) on the mitochondrial membrane. A decrease in the number of free HK molecules increased the chance of Bax binding to VDAC I, leading to increased Bax-mediated apoptosis. In the presence of a static MF, LLPS was marked inhibited and HK recruitment was reduced, resulting in an increased probability of HK binding to VDAC I and a decreased probability of Bax binding to VDAC I, thus reducing Bax-mediated apoptosis. Our findings revealed a new physical mechanism for understanding magnetobiological effects from the perspective of LLPS. In addition, these results show the potential applications of physical environments, such as MFs in this study, in the treatment of LLPS-related diseases., (Copyright © 2023 Wen-Juan Lin et al.)

Amyloid protein cross-seeding is a peculiar phenomenon of cross-spreading among different diseases. Unlike traditional infectious ones, diseases caused by amyloid protein cross-seeding are spread by misfolded proteins instead of pathogens. As a consequence of the interactions among misfolded heterologous proteins or polypeptides, amyloid protein cross-seeding is considered to be the crucial cause of overlapping pathological transmission between various protein misfolding disorders (PMDs) in multiple tissues and cells. Here, we briefly review the phenomenon of cross-seeding among amyloid proteins. As an interesting example worth mentioning, the potential links between the novel coronavirus pneumonia (COVID-19) and some neurodegenerative diseases might be related to the amyloid protein cross-seeding, thus may cause an undesirable trend in the incidence of PMDs around the world. We then summarize the theoretical models as well as the experimental techniques for studying amyloid protein cross-seeding. Finally, we conclude with an outlook on the challenges and opportunities for basic research in this field. Cross-seeding of amyloid opens up a new perspective in our understanding of the process of amyloidogenesis, which is crucial for the development of new treatments for diseases. It is therefore valuable but still challenging to explore the cross-seeding system of amyloid protein as well as to reveal the structural basis and the intricate processes.

Introduction: Diorhabda tarsalis Weise is an important insect pest of the Chinese licorice Glycyrrhiza uralensis Fisch. Behavior of the beetle, including host location, oviposition site selection, self-defense, and aggregation, were regulated by plant volatiles or insect pheromones. Aim: In this study, Identification of ORs and function research on orco were carried out, these could lead to the development of understand for olfaction mechanism in D. tarsalis . Methods: ORs were identified by PacBio RS II platform to sequence the antennas of adult D. tarsalis , the function of orco was explored by dsRNA interference. Results: 29 odorant receptor candidate genes of D. tarsalis were obtained, which code for 130-479 amino acids. Phylogenetic trees of olfactory receptors were constructed with 243 ORs from eight Coleoptera species. DtarORco , DtarOR7 and DtarOR26 are specifically expressed in the antenna, and the expression levels were significantly higher than other Dtar ORs in antenna, there were no differential expression between male and female beetles. An odorant coreceptor gene ( DtarORco ) has characteristics of an odorant receptor family member, the encoded mature protein has a predicted molecular weight of 53.898 kDa, dsRNA L4440 expression vectors were constructed and successfully transformed into ribonuclease III-deficient Escherichia coli strain HT115 DE3. After interference treatment, the relative expression level of DtarORco in D. tarsalis antennae significantly decreased and electrophysiological responses to host localization odor signals significantly decreased. At the same time, beetles lost the ability to locate hosts. Discussion: The research on its mechanism of olfaction may lead to the development of new control measures that are environmentally friendly., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen, Dewer, Wang, Tan, Liu and Shi.)

Staphylococcus aureus (S. aureus) infection, especially its drug-resistant bacterial infection, is a great challenge often faced by clinicians and patients, and it is also one of the most important threats to public health. Finding a safe and effective antibacterial agent is of great significance for the prevention and treatment of S. aureus infection. Lysozyme is known to have antibacterial effects against Gram-positive bacteria including S. aureus. Here, high-quality lysozyme with a purity of more than 99% and an activity of more than 60, 000 U/mg was prepared from egg white, which showed excellent antibacterial activity against three strains of S. aureus, especially against MRSA. Furthermore, an antibacterial cream loaded with lysozyme was prepared and tested in scald wound healing. The lysozyme-loaded cream exhibited the effect of preventing wound infection and promoting wound healing on scalds, and no toxicity was found in animal organs. Overall, lysozyme showed great application potential in the prevention and treatment of infections caused by S. aureus and scalded wound healing. The most remarkable discovery in this work is the unexpectedly powerful inhibitory effect of lysozyme on the drug-resistant bacterial, especially MRSA, which is usually very difficult to deal with using normal antibacterial drugs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In our study, the anti-quorum sensing (QS) activity of fermentation broth from TRM B-02, a bacterium isolated from Taklimakan desert, was investigated using the biosensor bioassay on Chromobacterium violaceum ATCC12472. TRM B-02 was 100% similar to Bacillus subtilis subsp. Inaquosorum KCTC 13429 (T) by genotypic and phenotypic analyses. Based on anti-QS activity tracking, six known amicoumacins, named as AI-77-H ( 1 ), AI-77-F ( 2 ), amicoumacin B ( 3 ), amicoumacin C ( 4 ), AI-77-C ( 5 ) and bacilosarcins D ( 6 ), were isolated and identified. Among them, compounds 1-3 exhibited a better inhibitory effect on C. violaceum ATCC12472. Further research suggested that compounds 1 - 3 could significantly down-regulate the expressions of violacein operon A (vioA), vioB, vioD and vioE and up-regulate vioC. Docking experiments indicated that compounds 1-3 may act as an inhibitor of violacein biosynthetic pathway competitively inhibiting the binding of flavin adenine dinucleotide (FAD) with the vioD enzyme.[Figure: see text].

Objective: To analyze the differences in the composition and abundance of gut microbiota between patients with active pulmonary tuberculosis and healthy controls, and to identify the specific bacteria as biomarkers to distinguish between the two groups. Methods: Patients with active pulmonary tuberculosis treated in three municipal designated tuberculosis medical institutions in Sichuan, Jiangsu and Shanghai from September 2017 to September 2019 were selected as the case group ( n =88), and the healthy people without a history of tuberculosis from the same regions were recruited as the control group ( n =62). The fecal samples of the two groups were detected by 16S rRNA gene sequencing, and the differences of gut microbiota diversity, community composition and relative abundance at phylum and genus level from the two groups were analyzed. The random forest method was used to construct a predictive model to assess whether the specific bacterial flora could be used as biomarkers to distinguish tuberculosis patients from healthy people. Results: The alpha diversity analysis showed that the species richness and evenness of gut microbiota in tuberculosis patients were significantly lower than those in healthy controls ( P <0.001). There was a statistically significant difference in the composition of microbiota between the two groups (Bray-Curtis distance, P <0.001). In the gut microbiota of tuberculosis patients, opportunistic pathogens were relatively enriched, while some of the beneficial bacteria that can produce short-chain fatty acids were less abundant. The discrimination accuracy of the random forest model composed of Lachnospira , Lachnospiraceae ND3007 group and Roseburia was 76.67%, with area under the curve (AUC) being 75.29% (95% CI : 0.661-0.845). Conclusion: There were differences in gut microbiota between patients with active pulmonary tuberculosis and healthy people, and specific bacterial flora showed the potential to be used as biomarkers to distinguish between the two groups.

Proteins are like miracle machines, playing important roles in living organisms. They perform vital biofunctions by further combining together and/or with other biomacromolecules to form assemblies or condensates such as membraneless organelles. Therefore, studying the self-assembly of biomacromolecules is of fundamental importance. In addition to their biological activities, protein assemblies also exhibit extra properties that enable them to achieve applications beyond their original functions. Herein, this study showed that in the presence of monosaccharides, ethylene glycols, and amino acids, β-lactoglobulin (β-LG) can form assemblies with specific structures, which were highly reproducible. The mechanism of the assembly process was studied through multi-scale observations and theoretical analysis, and it was found that the assembling all started from the formation of solute-rich liquid droplets via liquid-liquid phase separation (LLPS). These droplets then combined together to form condensates with elaborate structures, and the condensates finally evolved to form assemblies with various morphologies. Such a mechanism of the assembly is valuable for studying the assembly processes that frequently occur in living organisms. Detailed studies concerning the properties and applications of the obtained β-LG assemblies showed that the assemblies exhibited significantly better performances than the protein itself in terms of autofluorescence, antioxidant activity, and metal ion absorption, which indicates broad applications of these assemblies in bioimaging, biodetection, biodiagnosis, health maintenance, and pollution treatment. This study revealed that biomacromolecules, especially proteins, can be assembled via LLPS, and some unexpected application potentials could be found beyond their original biological functions.

Background: To investigate the effects of chronic restraint stress on cognition and the probable molecular mechanism in mice., Methods: In the current work, a restraining tube was used as a way to induce chronic stress in mice. The protein levels were determined with ELISA and western blot. A series of behavior tests, including the Morris water maze, elevated plus maze, open field test, and novel object recognition test, were also performed to examine the anxiety and the ability of learning and memory. Moreover, murine neuroblastoma N2a cells were used to confirm the findings from mice under chronic stress., Results: Decreased synaptic functions were impaired in chronic stress with the downregulation of PSD95, GluR-1, the neurotrophic factor BDNF, and immediate-onset genes Arc and Egr. Chronic restraint decreased the histone acetylation level in hippocampal neurons while HDAC2 was increased and was co-localized with glucocorticoid receptors. Moreover, chronic stress inhibited the PI3K/AKT signaling pathway and induced energy metabolism dysfunctions., Conclusion: This work examining the elevated levels of HDAC2 in the hippocampus may provide new insights and targets for drug development for treating many neurodegenerative diseases., Competing Interests: Conflict of interest: The authors state no conflict of interest., (© 2021 Jie Wu et al., published by De Gruyter.)

The self-assembly of biomacromolecules is an extremely important process. It is potentially useful in the fields of life science and materials science. To carry out the study on the self-assembly of proteins, it is necessary to find out the suitable self-assembly conditions, which have always been a challenging task in practice. Inspired by the screening technique in the field of protein crystallization, we proposed using the same screening technique for seeking suitable protein self-assembly conditions. Based on this consideration, we selected 5 proteins (β-lactoglobulin, hemoglobin, pepsin, lysozyme, α-chymotrypsinogen (II) A) together with 5 screening kits (Index TM , BML, Morpheus, JCSG, PEG/Ion Screen TM ) to investigate the performance of these crystallization screening techniques in order to discover new optimized conditions of protein self-assembly. The screens were all kept at 293 K for certain days, and were analyzed using optical microscope, scanning electron microscope, transmission electron microscope, atomic force microscope, fluorescence microscope, and atomic absorption spectroscope. The results demonstrated that the method of protein crystallization screening can be successfully applied in the screening of self-assembly conditions. This method is fast, high throughput, and easily implemented in an automated system, with a low protein consumption feature. These results suggested that such strategy can be applied to finding new conditions or forms in routine research of protein self-assembly. KEY POINTS: • Protein crystallization screening method is successfully applied in the screening of self-assembly conditions. • This screening method can be applied on various kinds of proteins and possess a feature of low protein consumption. • This screening method is fast, high throughput, and easily implemented in an automated system.

Even though Paranosema locustae is widely used in China as a biological agent for controlling grasshoppers, the mortality rate is initially quite low. This study sought to determine whether the simultaneous use of P. locustae and Beauveria bassiana would be a more effective control strategy. Additionally, changes in the intestinal microbial communities of migratory locusts infected with the two pathogens were analyzed to investigate the roles of gut microbes in pathogen-host interactions. The mortality rate of locusts inoculated with B. bassiana and P. locustae simultaneously was not significantly higher than expected, but the mortality rates of locusts inoculated with B. bassiana 3, 6, and 9 days after inoculation with P. locustae were significantly higher than if their effects were additive, indicating synergism. A MiSeq analysis found that Weissella was the most common bacterium, representing 41.48% and 51.62% of the total bacteria in the mid- and hindguts, respectively, and the bacterial declines were greatest during dual infections with B. bassiana and P. locustae. The appropriately timed combined application of P. locustae and B. bassiana was more effective against locusts than either treatment alone. Moreover, the combined inoculation of the two pathogens changed the gut microflora of locusts, indicating the potential relevancy of their synergistic effects on locust control., (© 2020 Institute of Zoology, Chinese Academy of Sciences.)

Background: Multidrug-resistant tuberculosis (MDR-TB) is on the rise in China. This study used a dynamic Markov model to predict the longitudinal trends of MDR-TB in China by 2050 and to assess the effects of alternative control measures., Methods: Eight states of tuberculosis transmission were set up in the Markov model using a hypothetical cohort of 100 000 people. The prevalence of MDR-TB and bacteriologically confirmed drug-susceptible tuberculosis (DS-TB + ) were simulated and MDR-TB was stratified into whether the disease was treated with the recommended regimen or not., Results: Without any intervention changes to current conditions, the prevalence of DS-TB + was projected to decline 67.7% by 2050, decreasing to 20 per 100 000 people, whereas that of MDR-TB was expected to triple to 58/100 000. Furthermore, 86.2% of the MDR-TB cases would be left untreated by the year of 2050. In the case where MDR-TB detection rate reaches 50% or 70% at 5% per year, the decline in prevalence of MDR-TB would be 25.9 and 36.2% respectively. In the case where treatment coverage was improved to 70% or 100% at 5% per year, MDR-TB prevalence in 2050 would decrease by 13.8 and 24.1%, respectively. If both detection rate and treatment coverage reach 70%, the prevalence of MDR-TB by 2050 would be reduced to 28/100 000 by a 51.7% reduction., Conclusions: MDR-TB, especially untreated MDR-TB, would rise rapidly under China's current MDR-TB control strategies. Interventions designed to promote effective detection and treatment of MDR-TB are imperative in the fights against MDR-TB epidemics.

Substantial harm to ecosystems from the use of chemical pesticides has led to an increasing interest in the use of biopesticides to control grasshoppers in rangelands, including China. One such potential biopesticide for control of grasshoppers is the fungus Paranosema locustae . In this study, the dynamics of aboveground natural enemies of grasshoppers and arthropod diversity 0-9 years after application of P. locustae were investigated in rangeland in Qinghai Plateau, China. We found that the number of species and of individuals of aboveground natural enemies increased by 17-250% and 40-126%, respectively, after spraying P. locustae , and that the main natural enemies showed three peaks after treatment. The conventional indices of species diversity (H') and evenness (J') increased by 11-267% and 13-171%, respectively, after treatment with P. locustae . The results showed the positive effects of P. locustae on aboveground natural enemies and biodiversity in an arthropod community in Chinese rangeland. Paranosema locustae is thought to be a safe biological control agent for grasshopper management in Northwestern China.

Insects such as locusts and grasshoppers can reduce the effectiveness of pathogens and parasites by adopting different defense strategies. We investigated the behavioral thermopreference of Locusta migratoria manilensis (Meyen) (Orthoptera: Acrididae) induced by the fungus Beauveria bassiana, and the impact this behavior had on the fungal mycosis under laboratory conditions. By basking in higher temperature locations, infected nymphs elevated their thoracic temperature to 30-32.6 °C, which is higher than the optimum temperature (25°C) for B. bassiana conidial germination and hyphal development. A minimum thermoregulation period of 3 h/day increased survival of infected locusts by 43.34%. The therapeutic effect decreased when thermoregulation was delayed after initial infection. The fungus grew and overcame the locusts as soon as the thermoregulation was interrupted, indicating that thermoregulation helped the insects to cope with infection but did not completely rid them of the fungus. A significant enhancement in the number of haemocytes was observed in infected thermoregulating locusts, reaching levels that were even higher than those observed in the controls. In contrast, haemocyte concentration was severely reduced in infected insects that did not thermoregulate. In infected non-thermoregulating locusts, the reduction in haemocyte number was accompanied by an increase in fungal blastospore concentration that was obvious in the haemolymph by day four. In contrast, no circulating blastospores were found in the haemolymph of infected thermoregulating locusts three days post-inoculation. We also examined the phagocytic activity of infected insects in vivo by using fluorescein isothiocyanate (FITC)-labelled silica beads. The proportion of beads that was engulfed by haemocytes in infected, thermoregulating insects was similar to that in the controls throughout the experiment, whereas the rate of phagocytosis in infected, non-thermoregulating insects progressively decreased after infection. These findings demonstrated that behavioural thermoregulation can adversely affect B. bassiana mycosis in infected L. migratoria manilensis, thereby limiting the development of lethal entomopathogenic fungi in locusts. This is apparently accomplished through an increase in the levels of haemocytes, leading to greater phagocytic activity under certain environmental conditions., Competing Interests: The authors have declared that no competing interests exist.

miR-362 is a recently discovered member of the microRNA family, and it modulates a variety of physical activities and plays an important role in the occurrence and development of many tumors. However, the biological functions of hsa-miR-362-5p in non-small-cell lung carcinoma (NSCLC) are unknown. Transwell assay and colony formation were used to determine the migration, invasion, and proliferation of NSCLC cells in vitro . A subcutaneous tumor model in nude mice was established to detect NSCLC tumor growth in vivo . The direct binding of miR-362 to the 3'UTR of Semaphorin 3A (Sema3A) was confirmed by luciferase reporter assay. In this study, we found that the level of miR-362 was higher in NSCLC tissues than in adjacent normal tissues and that the level of miR-362 expression was also elevated in five NSCLC cell lines (A549, 95-D, H1299, H292, and H460) relative to a human normal lung epithelial cell line (BEAS2B). Furthermore, miR-362 promoted NSCLC cell invasion, migration, and colony formation in vitro and tumor formation in vivo . Next, we identified the miR-362 target gene Sema3A, which is significantly correlated with metastasis. Sema3A expression was increased in normal tissues relative to NSCLC tissues. This result is consistent with the fact that miR-362 expression is negatively correlated with Sema3A expression in clinical tissue samples and indicated that miR-362 can regulate Sema3A expression in NSCLC cells and consequently affect NSCLC invasion, migration, and colony formation. Taken together, these findings on the newly identified miR-362/Sema3A axis elucidate the molecular mechanism of NSCLC invasion and migration and could lead to a potential therapeutic target in NSCLC treatment.

While the importance of protein N-glycosylation in cancer cell migration is well appreciated, the precise mechanisms by which N-acetylglucosaminyltransferase V (GnT-V) regulates cancer processes remain largely unknown. In the current study, we report that GnT-V-mediated N-glycosylation of CD147/basigin, a tumor-associated glycoprotein that carries β1,6-N-acetylglucosamine (β1,6-GlcNAc) glycans, is upregulated during TGF-β1-induced epithelial-to-mesenchymal transition (EMT), which correlates with tumor metastasis in patients with hepatocellular carcinoma (HCC). Interruption of β1,6-GlcNAc glycan modification of CD147/basigin decreased matrix metalloproteinase (MMP) expression in HCC cell lines and affected the interaction of CD147/basigin with integrin β1. These results reveal that β1,6-branched glycans modulate the biological function of CD147/basigin in HCC metastasis. Moreover, we showed that the PI3K/Akt pathway regulates GnT-V expression and that inhibition of GnT-V-mediated N-glycosylation suppressed PI3K signaling. In summary, β1,6-branched N-glycosylation affects the biological function of CD147/basigin and these findings provide a novel approach for the development of therapeutic strategies targeting metastasis. © 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland., (© 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.)

Activated hepatic stellate cells (HSCs) release pro-inflammatory and pro-fibrogenic factors. CXC chemokine-ligand-1 (CXCL1) is expressed on HSCs. We previously found that the CD147 is overexpressed in activated HSCs. In this study, we showed an important role of CD147 in promoting liver fibrosis by activating HSCs and upregulating expression of chemokines. Specifically, we found that CD147 specific deletion in HSCs mice alleviated CCl₄-induced liver fibrosis and inhibited HSCs activation. Overexpression of CD147 upregulated the secretion of CXCL1. Meanwhile, CXCL1 promoted HSCs activation through autocrine. Treating with PI3K/AKT inhibitor could effectively suppress CD147-induced CXCL1 expression. Taken together, these findings suggest that CD147 regulates CXCL1 release in HSCs by PI3K/AKT signaling. Inhibition of CD147 attenuates CCl₄-induced liver fibrosis and inflammation. Therefore, administration of targeting CD147 could be a promising therapeutic strategy in liver fibrosis., Competing Interests: The authors declare no conflict of interest.

Background: We compared the effects of the new David I operation and classical Bentall operation in the treatment of aortic root disease combined with aortic insufficiency., Methods: A total of 60 cases of patients with aortic root disease combined with aortic insufficiency diagnosed at our hospital from January 2010 to January 2016 were analyzed retrospectively, including 32 cases of aortic root aneurysm, 18 cases of aortic dissection, 5 cases of hypertension combined with atherosclerosis, 2 cases of retrogression, 2 cases of rheumatic heart disease and 1 case of Takayasu arteritis. Twenty-four cases that underwent the David I operation and 36 cases that underwent the Bentall operation were selected and their therapeutic effects were compared. The operation success rate, operation time, cardiopulmonary bypass time, cross-clamp time and blood infusion of both groups were compared; there were no significant differences (P>0.05)., Results: Two patients in the David I group and 3 patients in the Bentall operation group died of multiple organ dysfunction. The LVEDd and LVEF of both groups postoperation had no difference when compared with those parameters of before operation. The diameter of the valve annulus after the operation was shorter than before the operation. The severity of valve regurgitation of both groups had no difference. However, the ratio of severe regurgitation of the David I group increased and the mild regurgitation decreased. The incidence rate of complications of the David I group was significantly lower than that of the Bentall operation group. The differences were statistically significant (P<0.05)., Conclusions: Both David I operation and Bentall operation have better short-term and long-term effects in the treatment of aortic root disease when combined with aortic insufficiency; however, David I operation had less long-term complications.

Tissue calcification is a major cause of failure of bioprosthetic heart valves. Aim of this study was to examine whether surface heparin treatment of the decellularized porcine heart valve reduces tissue calcification. Fresh porcine aortic heart valves were dissected as tissue discs and divided into four groups. Group A: controls without treatment, Group B: decellularization only, Group C: decellularization and glutaraldehyde cross-linking, Group D: decellularization and glutaraldehyde cross-linking followed by surface heparin treatment. After implantation in New Zealand White rabbits for 60 days, the explanted heart valve discs from the different study groups underwent a series of histological examinations as well as determination of calcium content by the methyl thyme phenol blue colorimetric method. Results of the explanted heart valve discs for the Von Kossa staining demonstrated that in Group A the heart valve tissue was the most severely stained with black color, whereas in Group D there was hardly any area that was stained black after implantation indicating the least tissue calcification. Furthermore, the inflammatory cells identified by the Hematoxylin-eosin staining appeared to be the least in Group D. The average tissue calcium content was highest in Group A (0.197 ± 0.115 μmol mg -1 ), modest in Group B (0.113 ± 0.041 μmol mg -1 ), and Group C (0.089 ± 0.049 μmol mg -1 ), and the lowest in Group D (0.019 ± 0.019 μmol mg -1 , p < 0.05). These results suggest that surface heparin treatment tends to reduce tissue calcification of the dellellularized porcine heart valve in a rabbit intramuscular implantation model. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 400-405, 2017., (© 2015 Wiley Periodicals, Inc.)

Lung cancer is the most commonly diagnosed type of cancer worldwide. Although TRIM65 is an important protein involved in white matter lesion, the role of TRIM65 in human cancer remains less understood. Here, we reported that TRIM65 was significantly overexpressed in lung cancer tissues compared with adjacent normal lung tissues. Furthermore, TRIM65 expression was closely related to overall survival of patients with lung cancer. Knock down of TRIM65 in two lung cancer cell lines, SPC-A-1 and NCI-H358, resulted in a significant reduction in cell proliferation, migration, invasion and adhesion and a dramatic increase in G0-G1 phase arrest and apoptosis. In vivo tumorigenesis experiment also revealed that depletion of TRIM65 expression inhibited NCI-H358 cell growth. Moreover, based on gene set enrichment analysis (GSEA) with The Cancer Genome Atlas (TCGA) dataset, we found that TRIM65 was positive related to cell cycle, metastasis up and RHOA-REG pathways, which was further validated by RT-PCR and Western blot in TRIM65 knockdown lung cancer cells and indicated a possible mechanism underlying its effects on lung cancer. In summary, our study suggests that TRIM65 may work as an oncogene and a new effective therapeutic target for lung cancer treatment.

Ostrinia furnacalis (Guenée) and Helicoverpa armigera (Hübner) are the most important pests of maize in China. A laboratory study and a 2-year field study on the efficacy of transgenic maize expressing the Cry1Ac protein BT38 against O. furnacalis and H. armigera were performed. We found that the husks, kernels, and silks of BT38 showed significant efficacy against larvae of O. furnacalis and H. armigera. In the field, when neonate larvae of O. furnacalis and H. armigera were on plants at different growth stages and when levels of leaf-damage or number of damaged silks were used to score efficacy, we found that BT38 showed significant insecticidal efficacy against O. furnacalis and H. armigera, but the non-Bt maize did not show significant efficacy against either pest. These results suggest that the insecticidal efficacy of Bt maize expressing the Cry1Ac protein could be useful in the integrated pest management of these key maize pests.

Crops producing insecticidal crystal (Cry) proteins from the bacterium, Bacillus thuringiensis (Bt), are an important tool for managing lepidopteran pests on cotton and maize. However, the effects of these Bt crops on non-target organisms, especially natural enemies that provide biological control services, are required to be addressed in an environmental risk assessment. Amblyseius andersoni (Acari: Phytoseiidae) is a cosmopolitan predator of the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae), a significant pest of cotton and maize. Tri-trophic studies were conducted to assess the potential effects of Cry1Ac/Cry2Ab cotton and Cry1F maize on life history parameters (survival rate, development time, fecundity and egg hatching rate) of A. andersoni. We confirmed that these Bt crops have no effects on the biology of T. urticae and, in turn, that there were no differences in any of the life history parameters of A. andersoni when it fed on T. urticae feeding on Cry1Ac/Cry2Ab or non-Bt cotton and Cry1F or non-Bt maize. Use of a susceptible insect assay demonstrated that T. urticae contained biologically active Cry proteins. Cry proteins concentrations declined greatly as they moved from plants to herbivores to predators and protein concentration did not appear to be related to mite density. Free-choice experiments revealed that A. andersoni had no preference for Cry1Ac/Cry2Ab cotton or Cry1F maize-reared T. urticae compared with those reared on non-Bt cotton or maize. Collectively these results provide strong evidence that these crops can complement other integrated pest management tactics including biological control.

Locusts aggregate into bands of nymphs and swarms of adults that can pose a major threat to crop. Previous studies have shown that infection by the microsporidian parasite Paranosema locustae prevents locust aggregation behavior and we show that gut bacteria, which produce components of locust aggregation pheromones, are substantially reduced in locusts infected with P. locustae. We found that P. locustae could reduce the diversity, abundance and community composition of Locusta migratoria's gut bacteria. The parasite infection was also shown to interrupt the peroxidase activity of locust hindgut. Genome-wide expression analysis showed that the parasite infection suppressed peroxidase mRNA relative expression of locust hindgut, but had no effects on attacin expression and superoxide dismutase at 16 d post-inoculation with 20,000 P. locustae spores. Our findings reveal the mechanisms by which P. locustae impairs bacterial diversity and community structure of Locusta migratoria's gut bacteria.

Aim: To retrospectively evaluate our experience with the diagnosis and surgical resection of esophageal gastrointestinal stromal tumors (GISTs)., Methods: Between January 2003 and August 2014, five esophageal GIST cases were admitted to our hospital. In this study, the hospital records, surgery outcomes, tumor recurrence and survival of these patients were retrospectively reviewed., Results: The median age of the patients was 45.6 years (range: 12-62 years). Three patients presented with dysphagia, and one patient presented with chest discomfort. The remaining patient was asymptomatic. Four patients were diagnosed with esophageal GISTs by a preoperative endoscopic biopsy. Three patients underwent esophagectomy, and two patients underwent video-assisted thoracoscopic surgery. The mean operating time was 116 min (range: 95-148 min), and the mean blood loss was 176 mL (range: 30-300 mL). All tumors were completely resected. The mean length of postoperative hospital stay was 8.4 d (range: 6-12 d). All patients recovered and were discharged successfully. The median postoperative follow-up duration was 48 mo (range: 29-72 mo). One patient was diagnosed with recurrence, one patient was lost to follow-up, and three patients were asymptomatic and are currently being managed with close radiologic and clinical follow-up., Conclusion: Surgery is the standard, effective and successful treatment for esophageal GISTs. Long-term follow-up is required to monitor recurrence and metastasis.

The aim of this research was to determine chemical composition and repellent and insecticidal activities of the essential oil of Kaempferia galanga L. rhizomes against the booklouse, Liposcelis bostrychophila Badonnel, and to isolate insecticidal or repellent constituents from the oil. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry. Twenty-eight components of the oil were identified. The major compounds in the oil were ethyl-rho-methoxycinnamate (38.6%), ethyl cinnamate (23.2%), 1,8-cineole (11.5%), trans-cinnamaldehyde (5.3%), and borneol (5.2%). Based on bioactivity-guided fractionation, four active constituents were isolated from the oil and identified as 1,8-cineole, ethyl cinnamate, ethyl rho-methoxycinnamate, and trans-cinnamaldehyde. The essential oil exhibited contact toxicity against the booklouse with an LC50 value of 68.6 microg/cm2. Ethyl cinnamate (LC50 = 21.4 microg/cm2) exhibited stronger contact toxicity than ethyl rho-methoxycinnamate and trans-cinnamaldehyde (LC50 = 44.6 and 43.4 microg/cm2, respectively) while 1,8-cineole showed weak acute toxicity. The essential oil also possessed fumigant toxicity against the booklouse with a LC50 value of 1.5 mg/liter air. 1,8-Cineole and trans-cinnamaldehyde (LC50 = 1.1 and 1.3 mg/liter, respectively) possessed stronger fumigant toxicity against the booklouse than ethyl cinnamate and ethyl rho-methoxycinnamate (LC50 = 10.2 and 10.2 mg/liter air, respectively). trans-Cinnamaldehyde was strongly repellent to booklice, whereas ethyl cinnamate and ethyl rho-methoxycinnamate were weakly repellent and 1,8-cineole did not repel booklice. The results indicate that the essential oil and its constituent compounds have potential for development into natural insecticides or fumigants and repellents for control of insects in stored grains.

Background: Esophagectomy through cervico-thoraco-abdominal approach is a useful surgical technique in treating patients with esophageal cancer. However, the cervical reconstruction is also known to have a high rate of anastomotic leakage, as well as anastomotic stricture, intrathoracic stomach syndrome, reflux esophagitis and other complications, thereby influencing postoperative recovery and quality of life., Aims: The objective of this study was to investigate whether tubular stomach is superior to whole stomach in reducing anastomotic leakage for esophageal reconstruction through the cervico-thoraco-abdominal (3-field) approach., Methods: A total of 850 patients undergoing the 3-field esophagectomy were retrospectively included in this study and divided into a tubular stomach reconstruction group (Group A, n=453) and a whole stomach reconstruction group (Group B, n=397). All patients underwent esophagectomy through right thorax, left cervical part, abdominal triple incisions and done in esophageal reconstruction by hand-sewn two-layer anastomosis., Results: Results revealed that in comparison with whole stomach, esophageal reconstruction with tubular stomach had a lower incidence of anastomotic leakage (5.5 vs. 9.3%, P<0.05), less manifestation of intrathoracic syndrome (3.3 vs. 9.8%, P<0.001) and less occurence of reflux esophagitis (5.1 vs. 11.1%, P<0.01). However, for the incidence of anastomotic stricture, there was no significant difference between the two groups (9.3 vs. 9.8%)., Conclusions: This observation study suggests that for esophageal cancer patients undergoing the 3-field esophagectomy tubular stomach is better than whole stomach for esophageal reconstruction as reflected by a reduced postoperative anastomotic leakage, intrathoracic syndrome and reflux esophagitis.

In our screening program for insecticidal activity of the essential oils/extracts derived from some Chinese medicinal herbs and spices, garlic (Allium sativum L.) essential oil was found to possess strong insecticidal activity against overwintering adults of Cacopsylla chinensis Yang et Li (Hemiptera: Psyllidae). The commercial essential oil of A. sativum was analyzed by gas chromatography-mass spectrometry. Sixteen compounds, accounting for 97.44% of the total oil, were identified, and the main components of the essential oil of A. sativum were diallyl trisulfide (50.43%), diallyl disulfide (25.30%), diallyl sulfide (6.25%), diallyl tetrasulfide (4.03%), 1,2-dithiolane (3.12%), allyl methyl disulfide (3.07%), 1,3-dithiane (2.12%), and allyl methyl trisulfide (2.08%). The essential oil of A. sativum possessed contact toxicity against overwintering C. chinensis, with an LC50 value of 1.42 microg per adult. The two main constituent compounds, diallyl trisulfide and diallyl disulfide, exhibited strong acute toxicity against the overwintering C. chinensis, with LC50 values of 0.64 and 11.04 /g per adult, respectively.

Human vascular cells from saphenous veins have been used for cell seeding on the synthetic scaffolds for constructing tissue-engineered heart valve (TEHV). However, little is known about the seeding of human vascular cells on bovine pericardium, a potential natural scaffold for TEHV. This study was aimed to assess the basic in vitro and in vivo characteristics of the human vascular cells seeded on decellularized bovine pericardium. In vitro, bovine pericardium samples with cell seeding were inspected on day 7, 14, and 21 by histology, scanning electron microscopy, and immunohistochemistry. In vivo, experiments were performed in nude mice by bilateral dorsal incision for the implantation of decellularized bovine pericardium with and without cell seeding. Results demonstrated that a total of 8-10 × 10(6) cells were obtained within 4-5 wk by the primary co-culture, which were detected positive for von Willebrand factor, α-smooth muscle actin antibodies, and fibronectin, indicating the presence of endothelial cells, smooth muscle cells, and fibroblasts, respectively. In vitro, the seeded cells showed a steady increase of endothelial activity from day 1 to day 7 and remained stable until day 21. After 30 days of implantation in vivo, the cells on the decellularized bovine pericardium could differentiate directionally and show all the identities of human endothelial cells, smooth muscle cells, and fibroblasts. These results indicate that the human vascular cells from the saphenous vein are an optional cell source for seeding on decellularized bovine pericardium scaffold for constructing TEHV., (Copyright © 2012 Wiley Periodicals, Inc.)

Objective: To explore the associations between the single nucleotide polymorphism of human mismatch repair gene hMLH1 and the papillary thyroid carcinoma (PTC) in Chinese Han people., Methods: A hospital based 1:1 matched case-control study was carried out. The single nucleotide polymorphism (-93G > A, 1151T > A and 655A > G) for 204 pairs of cases with PTC as well as healthy controls was identified by PCR-RFLP, PCR-ASO and DNA sequencing., Results: With univariate analysis, we found that compared to 1151TT genotype, the TA genotype could increase the PTC risk marginally, with odds ratio (OR) of 2.15 (95%CI: 0.99 - 4.85); While the mutant genotype TA + AA could increase the PTC risk statistically significant, with OR of 2.15(95%CI: 1.02 - 4.69). With 2 x 4 cross-over study, we found that compared to -93GG and 1151TT genotypes, individuals with both -93GA + AA and 1151TA + AA could increase the PTC risk marginally, with OR of 2.50 (95%CI: 0.96 - 6.67); While, compared to 655AA and 1151TT genotypes, individuals with both 655AA and 1151TA + AA could increase the PTC risk statistically significant, with OR of 2.50 (95%CI: 1.02 - 4.73). Multivariate and conditional logistic regression analysis showed the genotype of 1151TA, the history of receiving CT diagnosis, the history of tumor, the negative life events and eating seafood frequently could increase the risk of PTC, with OR of 6.79 (95%CI: 3.18 - 14.49), 3.35 (95%CI: 1.93 - 5.80), 39.03 (95%CI: 3.70 - 41.60) and 3.98 (95%CI: 1.81 - 8.73); While, eating fruit frequently could decrease the PTC risk., Conclusion: The 1151TA + AA genotype, the history of receiving CT diagnosis, the history of tumor, the negative life events and eating seafood frequently were the risk factors of PTC, while eating fruit frequently was the protective factor.

Objective: To differentiate adrenal adenoma from metastasis in patients using perfusion computed tomography (PCT) imaging., Methods: Thirty-two patients with adrenal masses underwent first-pass PCT imaging. Of these patients, twenty-one were diagnosed with adrenal adenoma, and the others with metastases. Perfusion maps of blood volume (BV), blood flow (BF), mean transit time (MTT) and permeability surface-area production (PS) were generated with an Advantage Windows workstation using the CT perfusion 3.0 software (General Electric Medical Systems, Milwaukee, WI). Histopathologic sections immunostained for CD34 were quantitatively evaluated for microvessel density (MVD)., Results: The perfusion parameters such as BV, BF and PS were statistically significant different between the two groups, with adenomas showing higher mean BV (12.18 versus 3.86), BF (97.51 versus 45.99) and PS (21.73 versus 10.93) compared with metastases (p<0.05). For BV, a cutoff point of 7.30 was found to have a sensitivity of 95.2% and a specificity of 100% to differentiate between adenoma and metastasis. The sensitivity and specificity were 81.0 and 80.0%, respectively, for BF with a cutoff point of 71.96; and 85.7 and 86.7%, respectively, for PS with a cutoff point of 12.70 to differentiate adenoma and metastasis. A comparison of MVD counts from adenomas with those from metastases showed a significant difference (p<0.05). However, no significant differences were observed in the four perfusion parameters and MVD between lipid rich and lipid poor adenomas., Conclusion: PCT may be useful for evaluating the neovascularization of adrenal masses and differentiating adenoma from metastasis on the basis of PCT parameters. Adenomas show higher BV, BF and PS compared with metastases. According our data, the optimal threshold BV is 7.30, resulting in a sensitivity of 95.2% and a specificity of 100% for the differentiation of adenoma from metastasis. Adrenal adenomas have similar hemodynamic profiles, which are apparently independent of the lipid content of an adenoma., (Copyright 2009 Elsevier Ireland Ltd. All rights reserved.)

Objective: To explore the relationship of the genetic polymorphisms and the haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma (PTC) in Chinese Hans., Methods: A hospital based 1:1 matched case-control study was carried out. The polymorphisms for 204 pairs of PTC cases and healthy controls were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific oligonucleotide (PCR-ASO) assays., Results: (1) The PTC risk was marginally increased in the hMLH1 1151TA genotype, with odds ratio (OR) of 2.15 (95%CI: 0.99-4.85); the PTC risk was significantly increased in the mutant genotype 1151TA+AA, with OR of 2.15 (95%CI: 1.02-4.69); (2) The haplotypes of -93G, 1151A, 655A in the hMLH1 gene could increase the PTC risk, with OR of 2.67 (95%CI: 1.16-6.53, P=0.011), compared with the haplotype of -93G, 1151T, 655A; (3) Compared to 3124A, 2835G haplotype in hMSH3 gene, the 3124G, 2835A haplotype could increase the PTC risk marginally, with OR of 3.08 (95%CI: 0.92-13.25)., Conclusion: The 1151T/A polymorphism in hMLH1 was associated with PTC; both the haplotype of -93G, 1151A, 655A in hMLH1 and the 3124G, 2835A haplotype in hMSH3 were associated with PTC.

Objective: To understand the risk factors of primary angle-closure glaucoma., Methods: One to one matched case-control study was conducted in this study. One hundred and ninety two PACG cases and 192 controls, matched by age and gender, were collected from Department of Ophthalmology, Affiliated Hospital of Nantong University. All of the participants were investigated for their demographic information, behavioral habits, disease history, glaucoma family history and received clinical ocular examinations. The difference between these two groups was analyzed., Results: Several factors, including hypertension (OR = 2.004, P = 0.009), glaucoma family history (OR = 6.726, P = 0.003), presbyopia (OR = 3.192, P = 0.031), shallow anterior chamber (OR = 12.804, P = 0.000) and high cup-to-disc ratio (OR = 9.401, P = 0.007) were associated with PACG by multiply Logistic regression. The results did not support that smoking, drinking, myopia, diabetes mellitus and blood style were related to PACG., Conclusion: Follow up the populations with glaucoma family history, shallow anterior chamber or high cup-to-disc ratio are the main procedures for the decrease of incidence of PACG.

Cellular oxalate, widely distributed in many plants, is implicated to play important roles in various functions and is also known to affect food qualities adversely in fruits and vegetables. How oxalate is regulated in plants is currently not well understood. Glycolate oxidase (GLO) has long been considered as an important player in oxalate accumulation in plants. To gain further insight into the biochemical and molecular mechanisms, the possible roles of GLO in the process were studied. Drastically different levels of oxalate could be achieved by treating rice with various nitrogen forms (nitrate versus ammonium). While nitrate stimulated oxalate accumulation, ammonium reduced its level. Such treatments resulted in similar pattern changes for some other related organic acids, such as glycolate, oxaloacetate, and malate. By feeding plants with exogenous glycolate it was possible almost completely to restore the ammonium-decreased oxalate level. Under the two treatments few differences were observed for GLO mRNA levels, protein levels, and in vitro activities. Both K(m) for glycolate/glyoxylate and K(i) for oxalate remained almost the same for GLO purified from either nitrate- or ammonium-fed leaves. A further in vivo study, with transgenic plants carrying an estradiol-inducible GLO antisense gene, showed that, while the estradiol-induced antisense expression remarkably reduced both GLO protein levels and activities, oxalate levels were not significantly altered in the estradiol-treated transgenic plants. Taken together, it is suggested that oxalate accumulation and regulation is independent of GLO in rice leaves.

Objective: To study the relationship between presence of HPV DNA related sequence and cellular-immune responses in patients with condyloma acuminata (CA)., Methods: HPV DNA related sequences in biopsy material of 30 CA patients were determined by using Southern blot hybridization, and typed according to the restricted fragment length polymorphism (RFLP) analysis. At the same time, the T-cell subpopulations in peripheral blood of 30 patient with CA were also measured by using anti-CD3, CD4, CD8 monoclonal antibody labeled SERC direct rosette assay., Results: HPV DNA positive results were revealed in 19 of 30 patients with CA(63.3%). Among them, 3 cases possessed HPV 6 DNA homologous sequence (15.8%), 15 had HPV 11 DNA homologous sequence(78.9%), and only one case had a sequence homologous to HPV 16 DNA(5.2%). The percentages of CD3 and CD4 cells were significantly lower (P< 0.01) in CA patients than those in controls. However the percentage of CD8 cell was strikingly higher (P< 0.001) in CA patients than that in controls. Consequently the CD4/CD8 ratio was noticeably lower (P<0.001) in patients with CA than that in controls. The decrease of CD3 cell and the lowering of C D4/CD8 ratio were closely related to the positivity of HPV DNA homologous sequence (P =0.008 02, P=0.004 17). Conclusion The presence of HPV DNA in the patients with CA results in inhibitory effects on a series of cellular-immune responses, which may play an important role in the pathogenesis of CA.

ErbB-2 is overexpressed in several human cancers and conveys a transforming activity that is dependent on tyrosine kinase activity. Antibodies and T cells to ErbB-2 have been isolated from cancer patients, indicating ErbB-2 as a potential target of active vaccination. In this study, 3 mutant ErbB-2 DNA constructs encoding full-length, ErbB-2 proteins were tested as tumor vaccines. To eliminate tyrosine kinase activity, the ATP binding lysine residue 753 was substituted with alanine by replacing codon AAA with GCA in mutant ErbB-2A. To direct recombinant ErbB-2 to the cytoplasm where major histocompatibility complex (MHC) I peptide processing takes place, the endoplasmic reticulum (ER) signal sequence was deleted in cyt ErbB-2. The third construct cyt ErbB-2A contained cytoplasmic ErbB-2 with the K to A mutation. Expression of recombinant proteins was measured by flow cytometry in transfected murine mammary tumor cell line D2F2. Transmembrane ErbB-2 and ErbB-2A were readily detected. Cytoplasmic ErbB-2 and ErbB-2A were detected only after the transfected cells were incubated overnight with a proteasome inhibitor, indicating prompt degradation upon synthesis. ErbB-2 autophosphorylation was eliminated by the K to A mutation as demonstrated by Western blot analysis. Growth of ErbB-2-positive tumor in BALB/c mice was inhibited after vaccination with ErbB-2 or ErbB-2A, but not with cyt ErbB-2 or cyt ErbB-2A. ErbB-2A that is free of tyrosine kinase activity is a potential candidate for anticancer vaccination. The 3 mutant constructs should be useful tools to delineate the role of individual immune effector cell in ErbB-2-specific antitumor immunity and to develop strategies for enhancing such immunity.

Molecular changes associated with breast cancer progression were characterized using the MCF-10F cell series. MCF-10F was established from fibrous mastectomy tissue of a patient without detectable cancer. In vitro treatment of MCF-10F cells with benzo(a)pyrene resulted in a transformed subclone MCF-10F-BP1 (BP1). Transfection of clone BP1 with T24-Hras resulted in the tumorigenic line MCF-10F-BP1-Tras (BP1-Tras). Using flow cytometry, the expression of HLA I, ERBB-2 and MUC-1 was found to be comparable in 'normal' MCF-10F, transformed BP1 and tumorigenic BP1-Tras cells. Glycosylated mucin is elevated in BP1 but reduced in BP1-Tras cells. Using mRNA differential display analysis, cDNA profiles of the 'normal', transformed and tumorigenic cell lines were strikingly similar, yet distinct and elevated expression of several common cDNA fragments was detected in BP1 and BP1-Tras when compared with MCF-10F cells. These fragments were cloned and sequenced. The sequences of clones T1-360 and C4-310 are homologous to two reported EST cDNA clones from human fetal tissue and were further characterized. Elevated expression of the genes corresponding to clones T1-360 and C4-310 was verified using Northern blotting. High-level expression of these genes was also detected in the breast cancer cell line MCF-7 that was derived from the pleural effusion of a patient with advanced breast cancer. Therefore, specific molecular changes associated with breast cancer development were identified and may be indicators of neoplastic progression.