Lymphocyte-activation gene 3 (LAG-3), also known as CD223, is a 498-amino-acid type I transmembrane protein encoded by LAG-3 gene, which consists of extracellular, transmembrane and intracellular regions.LAG-3 negatively regulates T lymphocyte by binding extracellular domain to ligand, thus avoiding autoimmunitycaused by T cell over-activation. Like programmed cell death 1 (PD-1) and cytotoxic T lymphocyte antigen 4 (CTLA-4), LAG-3 is an important immune checkpoint in vivo and plays a balanced regulatory role in human immune system.Tumor cells escape the surveillance of the immune system by over-expressing LAG-3 ligand. With the development in research of immune checkpoints, LAG-3 has become a new generation of immunotherapy targets after PD-1 and CTLA-4. This article reviews the structure and function of LAG-3 and the application of its inhibitors in tumor immunotherapy, in order to provide reference for the further study of LAG-3.

AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246～386aa expressed in this library ,purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5?. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.

Objective The experiment was designed to investigate the effect of acellular nerve allografts on the functional recovery and reconstruction of the nerve-muscle structure of the sciatic nerve defect in rats. Methods Acellular nerve allograft was transferred into the defected rat sciatic nerve with 10mm long.The wet weight of tibialis anterior was weighed at 12 and 24 weeks postoperatively compared with control group.The conducted velocity of regenerated nerve and the effect of regenerated nerve on tibialis anterior were investigated by electrophysiologic test,and silver staining combined with AChE histochemical methods were used in the experiment separately. Results The wet weight of tibialis anterior and the conducted velocity of regenerated nerve in experimental group were similar to those in control group in 12 and 24 weeks after transplantation.The positive acetylcholinesterase(AChE)histochemical reaction was observed in the tibialis anterior at 12 weeks with deeper staining and located in the middle of tibialis anterior tidily at 24 weeks after operation.The regenerated nerve bundles and nerve terminals were found to grow into the motor end-plate of the tibialis anterior in silver staining combined with AChE staining in experiment group.Electromyogram showed that the regenerated nerve has innervated tibialis anterior already.Conclusion The results indicated that extracted nerve allografts as a bridge can promote the motor functional recovery and reconstruction of the nerve-muscle structure of the defected rat sciatic nerve.

Objective To detect the effect of oridonin(ORI)on the gene expression of human esophageal carcinoma cell SHEEC and to screen the tumor cell apoptosis target genes.Methods The gene expression of human esophageal carcinoma cell SHEEC without and with ORI induction for 1 hours and 8 hours were detected with microarray technique,respectively.The differentially expressed genes were identified and verified with fluorecent quantitative PCR.Results A total of 1011 genes showed up or down regulation more than twice after ORI induction(including 280 genes after 1 hour and 731 genes after 8 hours induction respectively).In these genes,17 genes with the top extent of up or down regulation were identified,which were involved in the cell signal transduction,transcription regulation,and cell apoptosis.These 17 differentially expressed genes were verified with real-time PCR,and 12 genes were statistically significant.Conclusion In the 12 differentially expressed genes with statistically significance,there may have tumor cell apoptosis target genes induced by ORI through mitochondrion route.

Objective To explore the distributions of genotypes of HPV infection in CIN and cervical carcinoma. Methods Cervical exfoliated cells were collected from 365 patients with abnormal cervical histology , and subjected to genotyping assay. Results The most prevalent HPV types were 16, 18,52, 58 and 33. The prevalence ratio of HPV 33,52,58 was signi cantly lower in squamous cell carcinoma. Multiple infections decreased from CIN II/III to cervical cancer. Conclusion Besides HPV 16/18, the 52/58/33 subtypes are also important in the development of cervical cancer.

The research and quality control of traditional Chinese medicine has meaningful importance, because it has influence not only in the health and treatment of patients, but also in the solid growth and development of phar-maceuticals companies. In some cases, for the complex of TCM, the common QC method on single or multi-target compounds can't really and truly disclose the quality of the Chinese materia medica. Therefore, a lot of researchers do plenty of works to make clear the effectiveness basis, to improve the quality and realize the modernization of TCM. All of these works close together with modern analysis and separation technology. In this article, a novel analy-sis technology-UltraPerformance Convergence Chromatography (UPC2) based its characters and applications should be introduced. It should be a helpful technology for the TCM researchers to facilitate the study and QC works.

Objective Recent researches indicate that Eph/ephrin signaling pathway is possibly related to adult neurogenesis after cerebral injury..With brief introduction of their structures and interactions,the review focus on their possible mechanism in adult neurogenesis.Methods The literatures between 2010 and 2015 were retrieved following online databases:PUBMED,ScienceDirect,Wanfang and CNKI database.The key words used in the reasearch were Eph,ephrin,cerebral injury,adult neurogenesisand so on.Results Totally 42 related articles were enrolled.And these papers discribed how researchers performed their experiments and further explained Eph/ephrin 's vital roles in adult neurogenesis.Conclusion Eph/ephrin signaling can influence adult neurogenesis after cerebral injury.positively or negatively.And Akt may be a downstream signaling molecule of Eph/ephrin that change the progress of adult neurogenesis.However,the detailed mechanisms remain to be further study.

Objective To observe the repairing effect of the acellular nerve allografts on the sciatic nerve gap of rat. Methods The acellular nerve allografts,treated by hypotonic-chemical detergent,were put on the 10 mm gap of the sciatic nerve in the rat.The action potential of the regenerated nerves was determined by the electrophysiologic method 13 weeks after operation.The morphology of the regenerated nerves was observed under light microscope and electron microscope,and the results were analyzed statistically. Results No inflammation and rejected reaction were found in the period of 13 weeks after operation in the operated and control groups.There was no significant difference in number of the regenerated nerve fibers,diameter of the axons,and the thickness of the regenerated myelinated nerve between the experimental group and control group.Conclusion The present results indicated that the acellular nerve allografts had good biocompatibility for the host rat in vivo and might as a bridge promote the regeneration of the injured sciatic nerve.;

BACKGROUND: There is a positive correlation between the serum cholesterol and osteoarthrosis incidence. However, it is still unclear whether the high-sugar and high-fat diet participates in the process of articular cartilage degeneration leading to osteoarthrosis, and the participation mechanism is unknown. OBJECTIVE: To investigate the changes in knee cartilage morphology of New Zealand white rabbits fed with high-sugar and high-fat diet and to explore the effect of high-sugar and high-fat diet on articular cartilage degeneration. DESIGN, TIME AND SETTING: The randomized controlled experiment was performed in Animal Laboratory, Xiangya Medical College from January to September 2008. MATERIALS: Forty healthy adult male New Zealand white rabbits, weighing about 2 kg, were randomly divided into 4 groups with 10 in each group. METHODS: Male New Zealand white rabbits were divided into four groups: ①normal diet group(NG), ②high-sugar diet (mixture of 37% cane sugar and 63% normal fodder) group(SG), ③high-fat diet (mixture of 20% lard and 80% normal fodder) group(FG), ④high-sugar and high-fat diet (mixture of 37% cane sugar, 10% lard and 53% normal fodder) group(SFG), MAIN OUTCOME MEASURES: Fasting blood samples were extracted every 4 weeks. The levels of blood sugar, triglyceride, cholesterol and insulin were detected, and body mass was recorded. The femoral condyle cartilage were dyed with toluidine blue, observed with transmit electron microscope after general observation of rabbit knee joints at the 28th weeks. RESULTS: ①Blood sugar, triglyceride, total cholesterol in FG and SFG were obviously higher than that in NG, the level of blood insulin in FG and SFG increased first, and then decreased significantly (P 0.05). ② By light microscope and transmit electron microscope observation, the FG and SFG demonstrated that tide line disappeared, cartilage cells were ranged in disorder, collagen fibers fractured, cartilage cells and a change in shape, such as shrinkage. These changes of morphology were not observed in SG. CONCLUSION: Long-term high-fat diet or high-sugar and high-fat diet may induce or aggravate the disorder of articular cartilage, suggested that it may take part in the development of osteoarthritis.