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 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 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.

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

Objective To investigate the factors influencing prognosis in surgical treatment of thoracolumbar spinal injury combined with spinal cord injury so as to assist in the clinical treatment.Methods A retrospective study was made on 77 cases of acute thoracolumbar spinal injury combined with spinal cord injury admitted from July 2005 to April 2011.There were 66 men and 11 women,aged 14-66 years (mean,36.5 years).Neurological performance evaluated using American Spinal Injury Association (ASIA) scale was grade A in 31 cases,grade B in 11 cases,grade C in 10 cases,and grade D in 25 cases.Potential factors affecting the prognosis of spinal cord injury were identified using univariate analysis and incorporated into the Logistic regression equation to filter out the main influencing factors.Results Fifteen cases dropped out at a mean 55.5-month follow-up (range,24-96 months) and follow-up rate was 81％.Univariate analysis selected fracture or dislocation types (P ＜ 0.01),combined injury (P ＜ 0.05),use of glucocorticoid within 8 hours postinjury (P ＜ 0.05),standard transport by medical professionals (P ＞ 0.1),complete injury (P ＜ 0.01),visiting time (P =0.055),cause of disease before operation (P ＜ 0.05) and canal encroachment rate (P ＜ 0.01) as predictors of neurologic recovery in spinal cord injury (P ＜ 0.1).Logistic regression analysis revealed significant prognostic factors were canal clearance (P ＜ 0.01) and use of steroids within 8 hours postinjury (P ＜ 0.05).Conclusion Glucocorticoid pulse therapy within 8 hours postinjury and adequate spinal cord decompression may effectively improve the neurologic prognosis in thoracolumbar spinal injury with spinal cord injury.

Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.