1.The usage of TCRV? gene and activation of PTK signal pathway in McAb costimulated PBLs and induction of in vitro apoptosis of hepatoma cells
Shulin HUANG ; Jibin ZHOU ; Weichun CHEN
Chinese Journal of Immunology 1985;0(05):-
Objective:To study the expression of TCRV? subfamily which specially recognize the hepatoma cell antigen and the apoptosis of hepatoma cell induced by McAb costimulated PBLs.Methods:The change of the phenotype of PBLs was studied by flow cell cytometry and the level of the expression of TCRV? was studied by RT-PCR and Southern blot,the PTK by western blot.The hypermicroscopic ultrastructure was observed through transmission electron microscope.Results:The level of CD3 and CD8 of PBLs was significantly increased after acted with hepatoma cells,while there was no change in CD4.The expression of TCRV?7 of PBLs was dramaticly increased and peaked at 4 days.PTK increased correspondently,to 58% compared with 11% in control.Besides anti-CD3 McAb induced lymphocyte apoptosis,the mediated apoptosis of hepatoma cells was found in the other three groups.Conclusion:TCRV?7 was the tumor antigen specific T cell receptor,and it activate the PTK signal pathway.The McAb activated lymphocytes initiated apoptosis in hepatoma cells.
2.The experimental study of the cytotoxicity to breast cancer cell line using modified T lymphocyte by TCRVpT.1 gene
Licai HU ; Shudong QIU ; Shulin HUANG
Chinese Journal of Immunology 1985;0(02):-
Objective: To investigate the influence of transfection of TCRVJ37.1 gene in cytotoxicity of PBMC of healthy donor to breast cancer cell line MCF-7/S. Methods :pcDN*31vp'1 packaged by lipofectamine were transfected to healthy donor's PBMC,Flow Cytometer Analysis and MTT Colorimetric Assay were used respectively to test the expression of pcDNA3.1VB7.1 gene and the cytotoxicity of PBMC before and after gene transfection of healthy donor to breast cancer cell line MCF-7/S. Results: The expression of TCRVJ37.1 gene after transfection was obviously higher.There was distinguish differences of cytotoxicity before and after gene transfection. Conclusion:The modified healthy donor's PBMC by TCR gene could have stronger cytotoxicity to breast cancer cell line.
3.Effects of telmisartan on the expressions of PPARs and adiponectin receptor2 in the liver tissue of rat with nonalcoholic steatohepatitis
Xiaoci HUANG ; Shulin JIANG ; Funa LIU
Acta Laboratorium Animalis Scientia Sinica 2017;25(3):289-294
Objective To explore the effects of telmisartan on expression of peroxisome proliferators PPARs activated receptors and adiponectin receptor 2 in rats with nonalcoholic steatohepatitis (NASH).Methods Forty male SD rats were randomized into normal-diet control group (NC,n=15),high fat-diet control group (FC,n=15),and high fat-diet with telmisartan group (FT,n =10).NC group was given standard diet and the other two groups were given high-fat diet.At the end of the 12th week,5 rats which were randomly selected from both the NC and FC groups were given euglycermic hyperinsulinemia clamp to see if fat-liver model of rats with insulin resistance was successfully induced,and rat livers were removed for pathological examination to determine the extents of NASH.Afterwards,rats in the FT group was given telmisartan (5 mg/kg·d) while rats in both the NC and FC groups were given the same volume of 0.9% saline solution by intragastric gavage for another 4 weeks.After glucose infusion rates (GIRs) were obtained by the euglycermic hyperinsulinemia clamp technique at the end of the 16th week,all rats were sacrificed and the body weight was recorded,and serum lipids,aminotransferases and fasting blood glucose were measured.The mRNA expressions of peroxisome proliferator activated receptors (PPARs),adiponectin receptor-2 and angiotensin II type-1 receptor in the liver tissue were assessed by semi-quantitative reverse transcriptase polymerase chain reactions.Results The expressions of PPARα,PPARγ and AdipoR2 mRNA in the liver tissue of FC group were decreased significantly compared with the NC group (P<0.01),and the expression of AT1R mRNA of the liver tissue in FC group was increased significantly compared with NC group (P<0.01).Compared with the FC group,the expressions of PPARα,PPARγ and AdipoR2 mRNA in the FT group were increased (P<0.01).Serum aminotransferases,lipids and fasting blood glucose level in the rats of FC group were increased significantly compared with rats of the NC group (P<0.01),and serum aminotransferases,lipids and fasting blood glucose level in the rats of FT group were greatly improved compared with the FC group.Conclusions Telmisartan can improve glucose and lipid metabolism,stop weight gain,decrease liver index,and alleviate steatosis and inflammation of NASH rats by improving insulin resistance.Telmisartan may play an effective role in the protection of rat liver with NASH.
4.Progress in infection pathway and intracellular trafficking of adenovirus.
Wenfeng ZHANG ; Hongwei SHAO ; Hua HEL ; Shulin HUANG
Chinese Journal of Biotechnology 2014;30(6):864-874
The research on intracellular trafficking of adenovirus has been described mainly through observations of subgroup C adenoviruses in transformed cell lines. The basic elements of the trafficking pathway include binding to receptors at the cell surface, internalization by endocytosis, lysis of the endosomal membrane, escape to the cytosol, intracellular trafficking along microtubules, nuclear pore docking, and viral genome translocation into the nucleus. More than 80% of the adenovirus genome is delivered to the nucleus in a highly efficient manner in approximately 1 h. However, exceptions to this trafficking pattern have been noted, including: variations based on target cell type, cell physiology, and adenovirus serotype. This review summarizes mechanism of adenovirus infection pathway and intracellular trafficking, providinging a foundation for the development of clinical adenoviral vector.
Adenoviridae
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physiology
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Cell Membrane
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virology
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Cell Nucleus
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virology
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Cytoplasm
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virology
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Endocytosis
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Endosomes
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virology
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Genetic Vectors
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Humans
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Microtubules
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Virus Internalization
5.Assistance of Eudragit S-100 on protein refolding
Zhifeng HUANG ; Shulin YANG ; Yi ZHANG ; Wenke FENG ; Xiaokun LI
Journal of China Pharmaceutical University 2009;40(3):273-278
Aim: To study the effect of Eudragit S-100, a pH-responsive polymer, on protein refolding level, using recombinant human keratinocyte growth factor-2 (rhKGF-2) as a model protein. Methods: The refolding of rh-KGF-2 was performed by directly diluting denatured rhKGF-2 into a refolding buffer containing different concentrations of Eudragit. The ability of Eudragit S-100 to enhance protein refolding level was investigated using MTT assay, reverse phase HPLC, fluorescence emission spectroscopy and circular dichroism spectroscopy. Results: The addition of Eudragit S-100 in the refolding buffer significantly increased the rhKGF-2 refolding yield to 71%, when dilution refolding was conducted at 0. 5 mg/mL rhKGF-2. The outcome from the refolding study showed possibility of a special interaction between rhKGF-2 and Eudragit, suggesting that the refolding-enhancing ability of Eudragit S-100 was due to this interaction between Eudragit S-100 and rhKGF-2. Mean while, the result showed that the concentration of urea was also an important factor for the optimization of the refolding in the presence of Eudragit. Conclusion: Eudragit S-100 can significantly increase the refolding level of rhKGF-2.
6.Preparation and Characterization of Butylaminopropyl Silica Monolithic Column for Capillary Electrochromatography
Fanggui YE ; Yanyan HAN ; Shun WANG ; Baojun HUANG ; Shulin ZHAO
Chinese Journal of Analytical Chemistry 2010;38(2):192-196
A novel silica monolithic stationary phase functionalized with butylaminopropyl ligands for capillary electrochromatography(CEC) has been presented. The monolithic capillary columns were prepared by a sol-gel) process and subsequent a chemical modification. The amino groups on the surface of the stationary phase are meant to generate a substantial anodic electroosmotic flow (EOF). The butyl and propyl groups provide) hydrophobic properties. To evaluate the column performance, effects of buffer pH and organic modifier content on the EOF and electrochromatographic retention behavior of alkylbenzenes, organic acids and anilines were investigated. The monolithic stationary phase exhibited reversed phase (RP) chromatographic behavior toward neutral solutes. The model organic acid anion solutes were separated by the mixed mode mechanism, which comprised RP interaction, weak anion-exchange, and electrophoresis. Basic compounds such as anilines) were well separated on the butylaminopropyl silica monolithic column without peak tailing.
7.Seasonal changes of glucose levels in Oncomelania hupensis
Ting LIU ; Chunlan HUANG ; Jinsun YANG ; Shulin ZHOU
Chinese Journal of Schistosomiasis Control 2014;(5):544-546
Objective To study the seasonal changes of glucose levels per unit soft tissue of Oncomelania hupensis. Meth-ods O. hupensis snails were collected from the beach of the Qingyi River in Wuhu City,Anhui Province from August 2012 to July 2013. They were kept in minus 80℃refrigerator immediately. The male snails were distinguished from female,and their soft tissues were collected separately after crushing their shells. The hexokinase method was used to measure the glucose concen-trations,and the results were analyzed statistically. Results The contents of glucose decreased from March to July. The lowest glucose content in the female was 1.87μg/mg in March,and that in male was 3.70μg/mg in July. Both of them increased from August and reached peak levels in September(♀=57.38μg/mg,♂=44.39μg/mg),and then gradually decreased from Octo-ber to next January and increased in February. Conclusion The contents of glucose have seasonal changes regularly in O. hu-pensis.
8.Biological effect of recognizing hepatoma antigen TCRV?7 subfamily after cloning and transgene
Weichun CHEN ; Yuexia LIN ; Shulin HUANG ; Al ET
Chinese Journal of Immunology 1985;0(03):-
Objective:To study the biological effects of TCR to hepatoma cell by transfection V?7 to lymphocytes. Methods:TCRV?7 gene was amplified by RT PCR and cloned to expression vector pLXSN. The recombinant was transferred into lymphocytes by Lipfectin Reagent transfection,then the lymphocytes were co cultured with hepatoma cells.The phenotype of lymphocytes was detected on the Flow Cytometry and the ultrastructure of the hepatoma cells was showed by electronic microscope.Results:The lymphocyte amount with TCRV?7 expressing in those being transfected was much more than those no transfection.Apoptosis appeared in the hepatoma cells.Conclusion:TCRV?7 subfamily can recognize hepatoma antigen and stimulate T cell.
9.Comparison of Several Methods and Their Progresses on Molecular Authentication of Lonicera japonica Thunb
Chao JIANG ; Luqi HUANG ; Yuan YUAN ; Shulin YU ; Min CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(8):1831-1839
Authentications of Chinese herbal medicine have a critical effect in Chinese clinical medicine. DNA molecular marker, as an important component for true or false authentication, is more and more widely used in iden-tification of Chinese medicinal materials. At the same time, many new methods for authentication of Chinese medici-nal materials are continuously emerging. But the systematically comparative analysis of these new methods is lack. The present study taking Lonicera japonica as an example, systematically compared principles, characteristics, ex-periment methods, detection time and the application scope of express sequence tag-simple sequence repeat (EST-SSR), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), allele-specific PCR (AS-PCR), DNA barcoding and loop-mediated isothermal amplification (LAMP), and put forward corresponding improve-ment opinions. This study can help to screen appropriate approach for rapid authentication of L. japonica and offer demonstrating to other Chinese herbal medicines.
10.Clinical Effect of Prostatil Combined with Diosmin on Chronic Prostatitis and Serum and Prostate Fluid Levels of MIP-2 and MIP-1α
Jian HUANG ; Hongwei DENG ; Dan CHEN ; Shulin YANG ; Ming SHI
Progress in Modern Biomedicine 2017;17(25):4920-4923
Objective:To explore the clinical effect of prostatil combined with diosmin on the elderly patients with chronic prostatitis (CP) and the macrophage inflammatory protein-2 (MIP-2) and macrophage inflammatory protein-lα (MIP-lα) in prostate fluid and serum.Methods:126 cases of elderly patients with CP in our hospital fiom January 2015 to September 2016 were selected and randomly divided into two groups.Prostatil combined with diosmin were provided to the patients in observation groups (63 cases) while the control group (63 cases) was treated by prostatil alone.The clinical effect,MIP-2,MIP-1α levels in the prostate fluid and serum before and after therapy as well as the incidence of adverse reactions were observed and compared between two groups.Results:At 12 weeks after treatment,the total effective rate of observation group was 93.7%,which was obviously higher than that of the control group (81.0%,P<0.05).The MIP-2 and MIP-1α levels in prostate fluid and serum of both groups at 12 weeks after therapy were significantly lower than those before therapy (P<0.01),which were significantly lower in the observation group than those of the control group at the same time (P<0.01).There was no significant difference in the incidence of adverse reactions between the two groups (P>0.05).Conclusion:Prostatil combined with diosmin could more safely and effectively improve the clinical efficacy in the treatment of elderly patients with CP/CPPS,which might be related to reduce the levels ofMIP-2,MIP-lα in serum and prostatic fluid.