1.Effect of SVCⅢ on the proliferation and apoptosis of human cutaneous T-cell lymphoma Hut-78 cells
Journal of Third Military Medical University 2003;0(15):-
Objective To explore the inhibition and apoptosis of the human cutaneous T-cell lymphoma cell lines Hut-78 by traditional Chinese medicine SVCⅢ. Methods After Hut-78 cells were treated with SVCⅢ of different concentration, the inhibition and apoptosis of Hut-78 cells was determined by MTT, agarose gel electrophoresis of DNA fragment and FACS. Results SVCⅢ could inhibit remarkably Hut-78 cells growth and DNA ladder was seen by agarose gel electrophoresis. The proliferation of Hut-78 cells were inhibited in G_1 stage by FACS. Conclusion SVCⅢ can promote growth retardation and apoptosis of human cutaneous T-cell lymphoma cell lines Hut-78, which suggests SVCⅢ has antineoplastic function.
2.Construction and identification of eukaryotic cell expression vector of human FHIT gene
Zhongwei TIAN ; Xiangfeng SONG ; Zhenhui PENG ;
Journal of Third Military Medical University 1984;0(02):-
Objective To construct eukaryotic cell expression vector of human frangible histone triad (FHIT) gene. Methods A 456 bp cDNA fragment was amplified from the total RNA of normal human thyroid tissue by RT PCR method and cloned into plasmid pcDNA3. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes Kpn Ⅰ and Bst XⅠ and sequenced by Sanger dideoxy mediated chain termination. The expression of FHIT gene was detected by immunocytochemical methods. Results The results showed that the cDNA fragment included 456 bp entire coding region. The recombinant eukaryotic cell expression vector of pcDNA3 FHIT was constructed, and the sequence of the insert was identical to the published sequence. MM96L cells transfected with the pcDNA3 FHIT plasmid expressed high level of Fhit protein in cytoplasm. Conclusion The recombinant plasmid pcDNA3 FHIT can provide a strong molecular tool for the studies of neoplasm pathogenesis.
3.Exploration on Experimental Teaching Mode of Medical Immunology
Xiangfeng SONG ; Guojun ZHANG ; Zhongwei TIAN ; Changen DUAN
Chinese Journal of Medical Education Research 2006;0(11):-
The exprimental teaching of medical immunology is the important constituent.We have explored the existing problems on course content,teaching method and experimental test way according to our practical teaching experience for the past few years.We have also made the preliminary attempt about reforming exprimental teaching of medical immunology.
4.Epigallocatechin gallate inhibits the proliferation and apoptosis of keratinocytes induced by interleukin-17
Dandan FU ; Hua HU ; Min SUN ; Min LI ; Zhongwei TIAN
Chinese Journal of Dermatology 2016;49(9):636-640
Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) against interleukin (IL)-17-induced injury to keratinocytes,and to explore its mechanism.Methods Some cultured HaCaT cells were divided into 3 groups to be treated with IL-17 alone at concentrations of 50,70,90 μg/L,respectively,with those receiving no treatment as the blank control group.Some HaCaT cells were divided into 5 groups:IL-17 group treated with 90 μg/L IL-17 alone,IL-17 + EGCG group treated with 90 μg/L IL-17 and 60 μmol/L EGCG,IL-17 + SP600125 group treated with 90 μg/L IL-17 and SP600125 (a JAK signaling pathway inhibitor),IL-17+ EGCG + anisomycin group treated with 90μg/L IL-17,60xmol/L EGCG and anisomycin (a Janus kinase signaling pathway activator),and blank control group receiving no treatment.After different durations of treatment,CCK-8 assay was performed to evaluate cellular proliferative activity,flow cytometry to detect cell apoptosis,enzyme-linked immunosorbent assay (ELISA) to measure expression levels of IL-6,IL-23 and IL-8,and Western-blot analysis to determine protein expressions of c-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK).Results IL-17 promoted cellular proliferation of HaCaT cells,and the proliferation rate,which was correlated with the concentration of IL-17,reached the maximum in the 90-μg/L IL-17 group (P < 0.05).EGCG at 60 μmol/L significantly inhibited cellular proliferation of,promoted apoptosis in,and reduced IL-6,IL-23 and IL-8 expressions in,HaCaT cells induced by 90 μg/L IL-17 (all P < 0.05).Compared with the IL-17 group,the IL-17 + EGCG group and IL-17 + SP600125 group both showed significantly decreased P-JNK expression,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P < 0.05).However,compared with the IL-17 + EGCG group,the IL-17 + EGCG + anisomycin group showed significantly increased protein expression of P-JNK,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P < 0.05).Conclusion EGCG protected against IL-17-induced injury to HaCaT cells,such as abnormal cell proliferation,apoptosis and inflammatory response,likely by inhibiting the JNK signaling pathway.
5.Fragile histidine triad(FHIT)gene promoter methylation and protein expression in mycosis fungoides
Zhongwei TIAN ; Xiangfeng SONG ; Min LI ; Zhenhui PENG
Chinese Journal of Dermatology 2010;43(1):44-46
Objective To detect the FHIT gene promoter methylation and protein expression in mycosis fungoides(MF).Methods Tissue specimens were collected from 48 patients with MF and 18 normal human controls.FHIT protein expression was determined by immunohistochemistry,and methylation status of FHIT gene by methylation-specific PCR.Results Abnormal methylation of FHIT gene was found in 26(54.2%)out of the 48 specimens.Thirty(63.5%)specimens of MF were negative for FHIT protein,which was observed in all the control specimens.The promoter methylation of FHIT was closely correlated with the protein expression of FHIT,but unrelated to the sex of,tumor staging or lymph node metastasis in patients with MF.Conclusion The FHIT gene promoter methylation may contribute to the inactivation and abnormal expression of FHIT protein in MF.
6.Effect of the exogenous fragile histidine triad(FHIT)gene on the proliferation and apoptosis of cutaneous carcinoma cell line A431
Xiangfeng SONG ; Zhongwei TIAN ; Dandan FU ; Xinling BI
Basic & Clinical Medicine 2006;0(04):-
Objective To investigate the effect of the exogenous fragile hisdidine triad(FHIT) gene on the proliferation and the apoptosis of cutaneous carcinoma cell line A431,and to explore the mechanism of tumor suppression by the FHIT gene.Methods The plasmids pcDNA3-FHIT and pcDNA3-vector were transfected into the cutaneous carcinoma cell line A431 without FHIT gene expression,and then the transfected cells were screened by G418 and the expression of FHIT was determined by the immunocytochemical staining technique.The effect of FHIT on the growth characteristics of cutaneous carcinoma cell line A431 was observed by MTT,colony forming test and flow cytometry.Results Stable FHIT gene expressing A431 cells were produced,the proliferation activity and colony forming capability of A431FHIT were suppressed,whereas the apoptosis was increased.All these differences between A431-FHIT cells and the two control groups of cutaneous carcinoma cells had statistical significance.Conclusion Transfecting the exogenous FHIT gene into cutaneous carcinoma cells line A431can suppress the proliferation of tumor cells,and can also induce apoptosis and cell cycle arrest.
8.Research advances of forensically lfuid/tissues identiifcation based on DNA methylation analysis
Yunshu JIA ; Zhongwei CAO ; Wenqiong ZHANG ; Chao XIAO ; Tian WEI ; Shaohua YI
Chinese Journal of Forensic Medicine 2016;31(6):591-594
Identiifcation of tissues/body lfuids in forensic science is important for criminal cases investigation such as crime scene reconstruction, conclude the character of crime. Recently, many researches of Epigenetic shows that tissue speciifc differentially methylated regions(tDMRs) have the ability to as a biomarker for identiifcation of tissues/body lfuids. In this paper, we reviewed the study progress and summarized the probability, advantage and disadvantage as well as application value and the development direction of the application of DNA methylation in the aspect of identifying the tissues/ body lfuids source, aiming at providing a reference for the related research and application.
9.Surgical treatment of inferior pole comminuted fractures of patella with new type ten-sion band
Bin SUN ; Zhishan ZHANG ; Fang ZHOU ; Yun TIAN ; Hongquan JI ; Yan GUO ; Yang LV ; Zhongwei YANG
Journal of Peking University(Health Sciences) 2015;(2):272-275
Objective:To study the effectiveness of inferior pole fracture of patella treating by the new tension band.Methods:From Dec.2011 to Dec.2013, 21 patients with inferior pole fracture of patella were treated with the new tension band which consisted of cannulated screw, titanium cable and shims. There were 21 patients[10 males, 11 females, the average age was 54 years(21 to 79)],of whom,all were“fell on knees”.Results:The average operation time was 89 min (57-197 min),the follow-up visits were done from 7-31 months ( average 18 months) , the bone healing time was from 8-12 weeks (average 10.5 weeks).The post operation assessment was done by Bostman score, from 20 -30 (average 27),10 excellent,and 11 good.No complication occurred.Conclusion:The new tension band is the effective treatment for inferior pole fracture of patella.The internal fixation is reliable, it is simple to operate, and patients can take exercises as early as possible.Therefore, the new tension band has a bet-ter clinical value.
10.The influence of post-void residual volume on prostate specific antigen in patients with benign prostatic hyperplasia
Feng LI ; Laikun TANG ; Zulin WANG ; Jiping YANG ; Li SONG ; Feng TIAN ; Zhongwei YU
Chinese Journal of Postgraduates of Medicine 2014;37(35):44-46
Objective To study the influence of post-void residual volume on prostate specific antigen (PSA) in patients with benign prostatic hyperplasia (BPH).Methods One hundred and sixty BPH patients who diagnosed by postoperative pathologic manifestations were selected.The patients were divided into 4 groups according to post-void residual volume with 40 cases each:non post-void residual volume (post-void residual volume < 60 ml) group,less post-void residual volume (60 ml≤post-void residual volume < 120 ml) group,middle post-void residual volume (120 ml ≤post-void residual volume < 180 ml) group,mass post-void residual volume (post-void residual volume ≥180 ml) group.The serum PSA levels of 4 groups were observed.Results The PSA level in mass post-void residual volume group [(8.52 ± 6.72) iμ g/L] was significantly higher than that in non post-void residual volume group [(5.26 ± 4.16) μ g/L] and less post-void residual volume group [(5.93 ± 5.20) μ g/L],there were statistical differences (P < 0.05),there was no statistical difference between non post-void residual volume group and less post-void residual volume group (P > 0.05).Conclusions The serum PSA level in BPH patients with non post-void residual volume has no change,while the serum PSA level in BPH patients with mass post-void residual volume is significantly elevated.