Objective To study cervical HPV infection in female patients with vulvar condyloma acuminatum(CA)from Shanghai area.MethodsExfoliated cells were obtained from cervices and vulva lesions of CA of 194 patients.respectively.HPV genotyping was carried out in cell samples using capture-hybridization method and gene chip techniques.Results HPV was detected in vulva lesions of all the 194 patients.Among them,74.2%(144/194)were positive for low risk(LR)-HPV,and 25.8%(50/194)for high risk(HR)-HPV.A single HPV genotype(6 or 11)was detected in 136(94.4%)patients with LR-HPV.and mixed genotypes of LR-HPV and HR-HPV in 46(86%)patients with HR.HPV.Of the 194 patients.85.6%(166/194)were complicated by cervical HPV infection.including 119(61.4%)cases of LR-HPV and 46(23.7%)cases of HR-HPV,In the case of HPV genotype.the consistence between CA lesions and cervix was 95.8%(159/166).The prevalence of LR-HPV declined sequentially from subtype 11 to 6 and 53,and that of HR-HPV from subtype 16 to 18 followed by 52,31,45 and 58.Conclusions There is a high rate of infection with HR-HPV in female patients with CA.and nearly one quarter of these patients are complicated by cervical HR-HPV infection.

Objective To investigate the protective effect of butyl flufenamate ointment against ultraviolet (UV)-induced skin damage,skin aging,and cutaneous squamous cell carcinoma (CSCC) in SKH-1 hairless mice.Methods A total of 128 mice were randomly and equally divided into four groups:UV group receiving UV irradiation only,butyl flufenamate ointment group and matrix cream group receiving UV irradiation after 30-minute pretreatment with topical butyl flufenamate ointment and matrix cream respectively,and blank control group receiving neither pretreatment nor irradiation.In the sunburn experiment (n =24),mice were exposed to single session of UV irradiation (1.5 minimal erythema doses (MEDs)),and 24 hours later,erythema and swelling response was observed,and skin tissue was obtained from the irradiated area on the back of mice followed by the determination of COX-2 expression using the streptavidin biotin peroxidase complex (SABC) method.To establish a photoaging (n =24) and CSCC (n =80) model,mice were exposed to four sessions of UV irradiation every week for 12 and 28 successive weeks respectively,with the irradiation dose starting at 0.9 MED and increasing gradually.After 12-week irradiation,skin tissue was resected from the back of photoaged mice and subjected to Masson staining for the evaluation of collagen changes as well as immunohistochemical analysis for the quantification of Bax,Bcl-2 and Caspase 3 expression.The initiation and progression of CSCC were observed in mice on a once-a-week basis from 12 to 28 weeks.SPSS 21.0 software was used for statistical analysis.One way analysis of variance was carried out for multiple-group comparisons of numerical data,Ridit analysis for the comparison of immunohistochemical staining intensity.Kaplan-Meier method and log-rank test were utilized for the comparison of tumor-free survival time.Results Both the degree of erythema and swelling response and expression level of COX-2 were significantly lower in the butyl flufenamate ointment group than in the other two UV-irradiated groups (all P ＜ 0.05).After 12-week irradiation,the butyl flufenamate ointment group showed milder degree of skin aging,together with higher density of collagen in dermis,weaker expression of Bcl-2 but stronger expression of Bax and Caspase 3,by comparison with the other two UV-irradiated groups (all P ＜ 0.05).During the 28 weeks of irradiation,the median tumor-free survival time was statistically longer in the butyl flufenamate ointment group than in the matrix cream group and UV group((25.0 ± 0.4) months vs.(24.0 ± 0.3) months and (23.0 ± 0.4) months,P ＜ 0.05 and 0.01 respectively).Conclusion Butyl flufenamate ointment has a certain photoprotective effect.

Animals ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Gene Rearrangement ; Genes, myc ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Translocation, Genetic ; Vincristine ; therapeutic use

Alzheimer's disease (AD) is a chronic neurodegenerative disorder and one of the earliest sings of AD is deficit in short term memory. Till now, the pathogenesis of AD has not been elucidated and the present one-drug-one-target paradigm of anti-AD-drug treatment seems not to be effective in clinic. Multi-target-directed anti-AD-drugs are those agents that may act on two or more targets implicated in AD. Based on the brief introduction of progress in the development of present anti-AD-drugs, the paper mainly focused on the advances in the field of multi-target-directed drug development both home and abroad, especially those studies on selective estrogen receptor modulators.
Alzheimer Disease ; drug therapy ; Animals ; Drug Combinations ; Drug Delivery Systems ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Indans ; therapeutic use ; Indoles ; therapeutic use ; Selective Estrogen Receptor Modulators ; therapeutic use ; Tacrine ; analogs & derivatives ; therapeutic use ; Thioctic Acid ; analogs & derivatives ; therapeutic use

Objective To establish a human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein. Methods HPV16E7 gene was amplified from CaSki cells using PCR and inserted into the eukaryotic expression plasmid pcDNA3.1. Then, the recombinant expression plasmid pcDNA3.1-HPV16E7 was transfected into HaCaT cells followed by G418 selection and identification by RT-PCR and Western blot. Results The recombinant eukaryotic expression plasmid pcDNA3.1-HPV16E7 was successfully identified by restriction enzyme digestion pattern and sequence analysis. Agarose gel electrophoresis of RT-PCR products detected the 297-bp fragment of HPV16E7 cDNA, and Western blot confirmed the stable expression of HPV16E7 protein. Conclusion A human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein is successfully established.

Objective To evaluate the effects and underlying mechanisms of crocin on glutamate-induced apoptosis of retinal ganglion cells (RGCs) by affecting extracellular calcium influx.Methods Primary rat retinal ganglion cells were isolated and stimulated with glutamate at concentrations of 0.1 mmol/L and 1 mmol/L for 24 h or 48 h,respectively,to establish apoptosis model of RGCs.Afterwards,crocin of different doses (0.1,1.0 and 3.0 μmol/L) was used to treat the glutamate-induced RGCs for 12 h;then cell apoptosis was detected by Annexin V-FITC/PI staining.The intracellular calcium concentration was determined by FIuo-3/AM fluorescent labeling.Western blot was used to examine the effect of crocin on Ca2+-mediated apoptotic signal molecules calpain and CaMKII.The mitochondrial membrane potential was detected by JC-1 staining and mitochondrial apoptosis-related signaling molecules Caspase-3,Caspase-9 and Bcl-2/Bax were evaluated by Western blot,respectively.Results In comparison with the untreated controls,the cell apoptosis of RGCs exposed to 0.1 mmol/L of glutamate for 24 h did not significantly change (P＞ 0.05).However,apoptosis rate of the cells reached (43.050 ± 2.616) ％ when the stimulation time lasted for 48 h and showed a significant increase (P＜0.01).Treatment with higher-dose glutamate (1 mmol/L) significantly increased apoptosis of RGCs at 24 h (46.450±1.061)％ and 48 h (45.500±3.253)％ compared with the controls (P＜0.01).RGCs were induced by 1 mmol/L of glutamate for 12 h,followed by the treatment with crocin at concentrations of 0.1,1.0 and 3.0 μmol/L,respectively.Each dose of crocin could significantly inhibit cell apoptosis in the dose-dependent manner (P＜0.01).In addition,crocin at 1.0 μmol/L blocked glutamate-induced extracellular calcium influx,inhibited the expression of calcium-dependent proteins Calpainl and CaMK Ⅱ.Moreover,crocin at the dose of 1.0 μmol/L also increased mitochondrial membrane potential,suppressed the expressions of Caspase-3 and Caspase-9,and elevated Bcl-2/Bax ratio.Cornclusion Crocin inhibits glutamate-induced apoptosis of retinal ganglion cells through suppressing extracellular calcium influx,thereby blocking calcium-dependent and mitochondria-dependent apoptosis signaling pathways.

Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.

The practical experiences with subject service in Medical Library of Chinese PLA were summarized in terms of the establishment of user information records, accumulation and summation of information, and construc-tion of information platform. Suggestions were put forward for the improvement of subject service in Medical Library of Chinese PLA, such as holding the big information concept in the big data era, using the biomedical data as the target of study, providing various kinds of service, integrating seamless, dynamic and interactive library service into user service. The fact that focus on artificial intelligence, computer, information and coomunication technologies is one of the future directions was pointed out.

Objective To investigate the local immune response in condyloma acuminatum treated with aminolevulinic acid-photodynamic therapy (ALA-PDT).Methods In vitro and in vivo studies were performed.A previously established keratinocyte cell line human papilloma virus (HPV) 16E7/HaCaT which stably expresses HPV16E7 protein was used in this study.Peripheral blood mononuclear cells (PBMCs) were separated from 10 healthy volunteers.After pretreatment with ALA-PDT,HPV16E7/HaCaT cells were cocultured with the PBMCs for 3 hours in a Transwell chamber followed by the observation of chemotactic migration of PBMCs.Tissue samples were obtained from the lesions of 10 patients with condyloma acuminatum before,and at 1,2,3 and 48 hours after the first session of ALA-PDT.Immunohistochemistry was conducted to determine the number of CD4+ T cells,CD8+ T cells and CD68+ macrophages as well as CD4/CD8 T-cell ratio in the tissue samples.Results After 3-hour coculture with HPV16E7/HaCaT cells pretreated by ALA-PDT,PBMCs showed apparent chemotactic migration.Immunohistochemistry revealed a statistical increase in the number of CD4+ T cells,CD8+ T cells and CD4/CD8 T-cell ratio at 48 hours (all P ＜ 0.05),as well as in the number of CD68+ macrophages at 3 hours and 48 hours (both P ＜ 0.05) after the first session of ALA-PDT.Conclusion ALA-PDT may induce local antiviral immune response in condyloma acuminatum.