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.

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.

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.

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.

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.

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.

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 .

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.

Objective To evaluate effects of downregulation of glucose?6?phosphate dehydrogenase(G6PD) expression on proliferation and cell cycle distribution of cutaneous squamous cell carcinoma(CSCC)cells. Methods Western blot analysis was performed to measure the protein expression of G6PD in normally cultured human HaCaT keratinocytes, SCL?1 and A431 CSCC cells. When A431 cells grew to 85%-90%confluence, a small interfering RNA (siRNA)targeting G6PD(G6PD?siRNA group)and a negative control siRNA(siRNA control group)were transfected into them separately, and untransfected A431 cells served as the untransfected group. CCK?8 assay was performed to evaluate proliferative activity of the A431 cells on days 0, 1, 2, 3 and 4 after transfection, Western blot analysis to measure G6PD, cyclin D1 and CDK4 protein expressions in A431 cells, and flow cytometry to analyze cell cycle distribution in A431 cells after 48 hours of additional culture. Results The protein expression of G6PD was significantly higher in normally cultured SCL?1 cells(0.308 ± 0.023)and A431 cells(0.643 ± 0.046)than in HaCaT cells(0.100 ± 0.019, both P<0.05), and significantly higher in A431 cells than in SCL?1 cells(P<0.05). The G6PD?siRNA group showed significantly decreased protein expressions of G6PD, cyclin D1 and CDK4(0.134 ± 0.027, 0.154 ± 0.017 and 0.166 ± 0.017, respectively)compared with the untransfected group(0.425 ± 0.029, 0.344 ± 0.024 and 0.330 ± 0.020 respectively)and siRNA control group(0.444 ± 0.033, 0.350 ± 0.027 and 0.348 ± 0.018 respectively) (all P<0.05). Besides, the G6PD?siRNA group showed significantly decreased cellular proliferative activity on days 1-4 compared with the siRNA control group and untransfected group(all P<0.001), while there were no significant differences between the untransfected group and siRNA control group at any of the time points (all P > 0.05). Compared with the untransfected group and siRNA control group, the G6PD?siRNA group showed significantly higher proportions of A431 cells in G0/G1 phase(both P < 0.001), but significantly lower proportions of A431 cells in S phase(both P<0.001). Conclusion G6PD may play important roles in the regulation of proliferation and cell cycle distribution of CSCC cells.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Neoplasm Invasiveness ; Skin Neoplasms ; metabolism ; pathology