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.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.
4.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.
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.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.
7.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.
8.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.
9.Effects of ursolic acid on interleukin-33 expression in HaCaT cells induced by interferon-γ
Hua HU ; Xiangfeng SONG ; Min SUN ; Dandan FU ; Min LI ; Zhongwei TIAN
Chinese Journal of Dermatology 2016;49(7):495-500
Objective To evaluate effects of ursolic acid (UA) on interleukin?33 (IL?33) expression in HaCaT cells induced by interferon?γ(IFN?γ), and to explore their mechanism. Methods Some HaCaT cells were treated with UA at different concentrations(0, 0.1, 1, 5, 10, 20, 40 and 80μmol/L)for 24, 48 and 72 hours separately. Then, methyl thiazolyl tetrazolium(MTT)assay was conducted to evaluate cell proliferative activity. A cell model of inflammation was established by culture of HaCaT cells with the presence of 200μg/L IFN?γ. Some HaCaT cells were classified into several groups to be treated with IFN?γ(200μg/L)and UA(10 and 15μmol/L)alone or in combination (firstly treated with IFN?γ followed by UA treatment), and those receiving no treatment served as the blank control group. Reverse transcription PCR (RT?PCR) was performed to detect mRNA expressions of IL?6 and IL?33, and Western?blot analysis to measure IL?33 protein expression after 12?hour culture. The expressions of extracelluar signal?regulated kinase 1/2(ERK1/2)and phosphorylated ERK1/2(p?ERK1/2)were also measured by Western?blot analysis after 5?and 60?minute treatments with IFN?γand UA alone or in combination. Results MTT assay showed that the treatments with 5-20μmol/L UA for 24 hours had no effects on cell proliferative activity, while 40-80μmol/L UA could significantly inhibit it at 24, 48 and 72 hours (all P < 0.05). Thus, 10 and 15 μmol/L were chosen as the concentrations of UA for further study. After the treatment with 200μg/L IFN?γ, there was a significant increase in the expressions of IL?33 mRNA(0.812 ± 0.036 vs. 0.412 ± 0.021), IL?6 mRNA(0.947 ± 0.091 vs. 0.595 ± 0.030)and IL?33 protein(1.317 ± 0.119 vs. 0.147 ± 0.036)in HaCaT cells compared with the blank control group(all P<0.05). Compared with the IFN?γgroup, the IFN?γ+10?μmol/L UA group and IFN?γ+15?μmol/L UA group both showed significantly decreased expressions of IL?33 mRNA(0.447 ± 0.042 and 0.438 ± 0.028 respectively, both P<0.05), IL?6 mRNA(0.437 ± 0.099 and 0.350 ± 0.075 respectively, both P<0.05)and IL?33 protein(0.923 ± 0.058 and 0.564 ± 0.113 respectively, both P<0.05). There were no significant differences in IL?33 mRNA expression between the IFN?γ+10?or 15?μmol/L UA group and blank control group(P>0.05), while IL?33 protein expression was significantly lower in the IFN?γ+15?μmol/L UA group than in the IFN?γ+10?μmol/L UA group(P<0.05). The p?ERK1/2 protein expression significantly increased in HaCaT cells treated with IFN?γ for 5 and 60 minutes compared with the blank control group, but significantly decreased in the IFN?γ+15?μmol/L UA group compared with the IFN?γgroup(0.458 ± 0.053 vs. 0.941 ± 0.042 at 5 minutes, 0.302 ± 0.054 vs. 0.509 ± 0.032 at 60 minutes, both P < 0.05). However, no significant differences were observed in the total ERK1/2 protein expression between the IFN?γ+15?μmol/L UA group and IFN?γgroup at 5 or 60 minutes. Conclusion UA can suppress IL?33 expression in HaCaT cells induced by IFN?γ, likely by regulating expressions of the ERK signaling pathway?related proteins.
10.Significance of urodynamics detection in benign prostatic hyperplasia patients with cerebral vascular accident
Feng LI ; Fang WANG ; Laikun TANG ; Zulin WANG ; Jiping YANG ; Li SONG ; Feng TIAN ; Zhongwei YU
Chinese Journal of Postgraduates of Medicine 2016;39(8):727-729
Objective To explore the significance of urodynamics detection in benign prostatic hyperplasia (BPH) patients with cerebral vascular accident. Methods The results of urodynamics were detected in 60 BPH patients with cerebral vascular accident (research group) and 60 simple BPH patients (control group), and the results were compared. Results The levels of bladder compliance (BC), maximum bladder capacity (MCC), maximum urinary flow rate (Qmax), detrusor pressure before voiding (Pdet), maximal urethral closure pressure (MUCP), functional urethral length (FUL) in research group were significantly lower than those in control group:(25.0 ± 4.4)ml/cmH2O (1 cmH2O=0.098 kPa) vs. (34.0 ± 5.9) ml/cmH2O, (203.0 ± 16.8) ml vs. (256.0 ± 20.3) ml, (6.70 ± 0.25) ml/s vs. (11.10 ± 0.43) ml/s, (81.0 ± 13.6) cmH2O vs. (108.0 ± 20.2) cmH2O, (60.0 ± 9.8) cmH2O vs. (88.0 ± 12.2) cmH2O, (3.0 ± 0.5) cm vs. (5.2 ± 0.8) cm, P < 0.05. The level of postvoid residual volume (PRV) in research group was significantly higher than that in control group:(83.0 ± 9.4) ml vs. (48.0 ± 8.3) ml, P<0.05. The rate of low compliance bladder and unstable bladder in research group were significantly higher than that in control group:63.3%(38/60) vs. 25.0%(15/60), 66.7%(40/60) vs. 28.3%(17/60), P<0.01. Conclusions Cerebral vascular accident can increase the bladder dysfunction of patients with BPH .