Objective In order to study the pathogenesis of human papillomavirus(HPV) and seek for a therapeutic approach of the diseases caused by HPV, the construction of HPV18 E6E7 antisense RNA expressing recombinants was studied. Methods We amplified the HPV18 E6E7 816bp by PCR with HPV18 plasmid DNA as the template. pLNSX retroviruses were used as vectors,the HPV18 E6E7 retrovirus recombinants were constructed. And then the recombinants were cleaved with restriction endonuclease and hybridized with Southern blot for identifying the inserting direction and special check respectively. Results and conclusion The HPV18 E6E7 antisense RNA retrovirus expressing recombinants were screened and obtained,which had laid the foundation of studying the function of E6E7 genes further and explore whether the antisense technique can adjust and control the expression of E6E7 genes.

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.

Objective To explore the role of IL-18 in the pathogenesis of chronic eczema. Methods Twenty-seven patients with chronic eczema were enrolled into this study along with 12 normal human controls. The severity of eczema was evaluated by eczema area and severity index (EASI) in patients. Skin specimens and vein blood samples were obtained from all the subjects. Reverse-transcription PCR was performed to detect the mRNA expression of IL-18 and IFN-γ in the skin tissue, and enzyme linked immunosorbent assay (ELISA) to measure the protein expression of IL-18 and IFN-γ in the sera of these subjects. Results The mRNA expression level in patients and controls was 1.04±0.29 pg/mL and 0.52±0.15 pg/mL for IL-18, respectively, 0.96±0.34 pg/mL and 0.47±0.12 pg/mL for IFN-γ, respectively; a significant increase was observed in the mRNA expression level of both IFN-γ and IL-18 in the patients than in the controls (both P<0.01). Moreover, the mRNA expression level of both IFN-γ and IL-18 positively correlated with the severity of eczema in patients (r=0.737, 0.883, both P<0.01). The protein expression level of IL-18 and IFN-γ was 475.8±59.4 pg/mL and 10.1±7.0 pg/mL, respectively, in the patients, 123.6 ±29.5 pg/mL and 11.1±3.4 pg/mL, respectively, in the controls; a statistical difference was observed in the protein expression level of IL-18 (P<0.01), but not in that of IFN-γ(P>0.01), between the patients and controls. No significant correlation was observed betweenthe serum level of IL-18 or IFN-γ and sererity of eczema in the patients (both P>0.01). Conclusions IL-18 may be involved in the pathogenesis of chronic eczema. Also, in local lesions, IL-18 seems to correlate with the induction of production of Th1 type cytokines, such as IFN-γ which could subsequently mediate hypersensitivity response.

Objective To investigate the expression of survivin in keloid, and its significance in the development of keloid. Methods Tissue samples were obtained from 25 patients with keloid (12 males and 13 females, aging from 4 to 44 years with a disease course of 1-18 years), and 15 normal skin samples obtained from surgical operation served as the controls. Streptavidin-biotin-peroxidase complex (SABC) method was applied to detect the expression of survivin in these samples. Results No expressin of survivin was observed in normal controls, while it was expressed in 80.0% (20/25) of the keloid samples with the predominant distribution in fibroblasts and vascular endothelial cells. The positivity rates of survivin were 57.14% (4/7), 81.81% (9/11) and 100% (7/7) in tissues of low-grade, moderate-grade and high-grade keloid, respectively, with no significant difference among the three groups (P = 0.133 ). Similarly, no signifi- cant difference was observed in the positivity rate of survivin between recurrent patients and untreated patients (90.91% (10/11 ) vs 71.43% (10/14), P = 0.341 ). Conclusion Survivin might play a role in the development of keloid.

Animals ; Apoptosis ; physiology ; Carcinoma, Squamous Cell ; drug therapy ; physiopathology ; prevention & control ; Cyclooxygenase 2 ; Cyclooxygenase 2 Inhibitors ; Cyclooxygenase Inhibitors ; therapeutic use ; Head and Neck Neoplasms ; drug therapy ; physiopathology ; prevention & control ; Humans ; Membrane Proteins ; Neoplasm Invasiveness ; Neoplasm Metastasis ; drug therapy ; Neovascularization, Pathologic ; drug therapy ; metabolism ; Prostaglandin-Endoperoxide Synthases ; metabolism

Background and purpose:Smac/DIABLO was the only apoptosis-related protein that could inhibit IAPs directly and simultaneously.The four amino-residual AVPI(Ala-Val-Pro-lie)in its N-terminal was the very important domain that could stimulate apoptosis.This study investigated the effect of synthetic Smac peptide (SmacN7) on chemotherapy sensitivity of bladder cancer cells.Methods:SmacN7 penetratin peptide was synthesized and delivered into T24 cells.MTT assay was adopted to evaluate the viability of T24 cells induced by low-dosage of MMC. Flow cytometry was applied to analyze the proportion of apoptosis and Western blot was used to detect the expression of XIAP and caspase-3;The activity of caspase-3 was measured and the effect of SmacN7 combined with MMC on T24 cell lines was also determined.Results:SmacN7 penetratin peptide could successfully interact with endogenous XIAP and increase the proportions of apoptosis of T24 cell lines induced by low-dosage of MMC in a dose-and time- dependent manner.An obvious down-regulation of XIAP expression and up-regulation of caspase-3 was identified by Western blot.The activity of caspase-3 in experimental group was significantly increased as compared with that in the control group;Combining the treatment with SmacN7 penetratin peptide,the viability of T24 cells decreased to 55% and 72.7% in 24 hrs and 48 hrs respectively.Conclusion:SmacN7 penetratin peptide could act as a cell-permeable IAP inhibitor,inhibit the proliferation,induce apoptosis and enhance the chemo-sensitivity of bladder cancer cells to MMC. When combined with chemotherapy,it may be a very promising strategy for bladder cancer therapy.

Objective To investigate the expression and secretion of mice DNaseI gene plasmid transfected into bone marrow (BM-MSCs) mesenchymal stem cells. Methods The plasmids of mouse DNaseI gene had been transfected into the BM-MSCs of mice by liposomes. The expression of DNaseI gene in the BM-MSCs was detected by western blotting and the DNaseI activity was measured by DNA-methyl green substrate colorimetry. Results About 30% BM -MSCs were transfected with mice plasmid DNaseI gene, DNaseI was expressed in the transfected BM-MSCs and active DNaseI could be detected in the supernatant of cell culture. Conclusion The mice DNaseI gene plasmid can be transfected into mice BM -MSCs by liposomes and DNaseI gene can be expressed by the transfected BM-MSCs and active DNaseI can be secreted. This may provide potential target for the treatment of SLE.

Brown-Sequard Syndrome ; etiology ; Cervical Vertebrae ; Humans ; Intervertebral Disc Displacement ; complications ; surgery ; Male ; Middle Aged

Objective To investigate the correlation between QRS amplitudes and left ventricular wall thickness in autopsy specimens of elderly men.Methods The data of autopsy cases in our hospital since 1990 were retrospectively analyzed.The cases with QRS duration≥0.12 s and the pacing electrocardiogram were excluded.QRS amplitudes of standard 12-lead electrocardiography in 3 months before death were measured and the correlation between QRS amplitudes and left ventricular wall thickness was analyzed in the elderly men.Results Correlations were found between the amplitudes of the R waves in leads V5 ,V6, Ⅰ ,aVL[(1.1±0.7) mV, (0.95±0.6) mV, (0.44±0.3)mV and(0.35±0.3)mV] and left ventricular wall thickness[(13.6±5.4)mm;r=0.22,0.14,0.22,0.23,all P<0.05], and between the combination of QRS amplitudes SV1 +RV5 or RV6(1.9±1.2) mV] and left ventrieular wall thickness [(13.8± 5.4) mm; r = 0.23, P < 0.05].The correlationbetween the combination of QRS amplitudes (SV1 + RV5 or RV6 ) and left ventricular wall thickness was the strongest in 60-79 years old cases (r=0.48, P<0.01) ,and was decreased in 80-89 years old cases (r= 0.23, P<0.05).There was no correlation between the combination of QRS amplitudes (SV1+RV5or RV6) and left ventricular wall thickness in 90-101 years old cases (r= 0.03, P> 0.05).Conclusions Electrocardiogram is a reliable method for diagnosis of left ventricular hypertrophy in elderly men aged < 90 years.

Sulfate-reducing bacteria (SRB) are widespread in the environment. SRB are obligate anaerobes and capable of dissimilatory reduction of sulfate. SRB have application prospects in the control of environmental pollution due to that many pollutants can be removed by SRB. The biological characteristics and metabolic mechanisms of SRB are introduced, and the application of SRB in the treatment of environmental pollution is described in this paper. The research progress of Cr(Ⅵ ) reduction and Cr(Ⅵ ) removal from wastewater by SRB is reviewed, and future direction of research on the control of Cr(Ⅵ ) pollution by SRB is also analysed.