Objective To study the chemical composition of traditional Chinese medicine Xiasangju compound extract.MethodsThe ethanol extract of different organic solvent extraction,silica gel column chromatography,chromatography separation and purification technology for the separation of compounds,the structures were elucidated by chemical and spectral data.ResultsSix fiavonoid components were isolated from the ethyl acetate extract,respectively, kaempferol( Ⅰ ),quercetin( Ⅱ ),luteolin( Ⅲ ),quercetin-3-O-β-D-glucoside (Ⅳ),luteolin-7-O-β-D-glucoside ( Ⅴ ) and rutin (Ⅵ).ConclusionThe obtained compounds were isolated from compound Xiasangju for the first time.

Objective:To observe the clinical effects of targeted percutanous ozone ablation on lumbar disc herniation (LDH) patients with high intensity zone (HIZ) in lumbar disc annulus fibrosus on MRI T2 weighted imaging.Methods:136 LDH patients with HIZ in lumbar disc annulus fibrosus on MRI T2 were divided into two groups according to therapy methods.In group A,75 patients were injected with 2 ～5 mL of 40 μg/mL mixture of O3 and O2 after targeted percutanous puncturing under the guidance of X-Ray machine with C-type arm.In group B,61 patients were treated with conservative treatment.MacNab score criterion and Oswestry disability index (ODI) were used in assessment of the efficacy.Results:Except 24 patients,all the other cases were followed up for 18 ～ 44 months.At the postoperative 1st,2nd,3rd,6th,9th,12th and 18th month,according to MacNab score criterion,the effective rates were respectively 88.00 ％,90.67 ％,93.33 ％,89.39 ％,84.85 ％,78.13 ％ and 73.44 ％ in group A and respectively 68.85 ％,62.30 ％ 55.74 ％,61.82 ％,58.12 ％,54.17 ％ and 47.92 ％ in group B.There were significant differences between two groups at the same time point (P＜0.05).At the postoperative 12th and 18th month,ODI was lower in group A,and there was no significant difference between two time points (P＞0.05).But it was significant different with that preoperatively and in group B at the same time point (P＜0.05).Conclusion:Targeted percutanous ozone ablation is an effective method with stable clinical efficacy in treating LDH with HIZ in lumbar disc annulus fibrosus on MRI T2 weighted imaging.

Aim To investigate the molecular mechanisms of differentiation in human leukemia HL-60 cells induced by diallyl disulfide(DADS)using suppression subtractive hybridization(SSH).Methods In our privious study,the subtractive cDNA library was constructed successfully and efficiently. 30 clones were randomly analyed with restriction enzyme.The inserts of cDNAs were analyzed by restrictive enzyme EcoR I.Positive clones were sequenced and the homology of resulting cDNA sequences were analyzed through bioinformatics software Blastn.Results 18 clones contained 100～600 bp cDNA inserts.10 differantiation genes were obtained and involved in cell cycle,signal transduction,metabolism and RNA binding.And 3 of 10 genes,p21,STAT1 and CAMTA1 were up-regulated and detected by RT-PCR,the results matched with SSH.Conclusion sThere are tight correlation between the differentiation induced by DADS and three-upregulated gene:p21,STAT1 and CAMTA1.

Objective To observe the anatomic relation of the anterior inferior cerebellar artery and the vestibulocochlear nerve. Methods The anterior inferior cerebellar artery and branches related with the vestibulocochlear nerve in the cerebellopontine angle were examined with the operating microscopy in 20 adult head cadavers fixed with formalin. Results The incidence of the anterior inferior cerebellar artery, recurrent perforating artery and anterior inferior cerebellar artery loop was 100percent.The 80 percent recurrent perforating arteries passed between the facial nerve and the vestibulocochlear nerve. The 95 percent vestibulocochlear nerves had contact with the anterior inferior cerebellar artery or its branches. Among them, The 50 percent had two or more branches contacted with the vestibulocochlear nerves.Conclusion The result will supply useful anatomic data for the surgery in the cerebellopontine angle.

Aim To investigate JAKs/STATs signal transduction change in HL-60 cells differentiation induced by diallyl disulfide(DADS)and molecular mechanism regulating the differentiation.Methods After incubation of HL-60 cells with DADS or AG490(50 ?mol?L -1),the cell differentiation indexes were observed by cytomorphology, NBT reduction ability assay,cell myeloid differentiation antigen CD11b by flow cytometry. Kinase activity of JAKs/STATs was tested by western-blotting and expressions of nucleus transcription genes stats,c-myc,c-fos,c-jun were detected by immumocyte chemistry method.Results Cell differentiation index changes indicated that HL-60 cells were induced differentiation toward granulocytic lineage by DADS, Western blot test demonstrated that constitive phosphorylation of Jak1,stat3 kinase was suppressed. Stat3,c-myc gene expression decreased and c-fos, c-jun gene expression increased in HL-60 cells treated with DADS through immunocyte chemistry.Conclusion Inhibition of phosphative Jak1, Stat3 was involved in HL-60 cells differentiation induced by DADS, its molecular mechanism might be related to modulation of gene expression associated proliferation and differentiation,and inhibition of DNA systhesis, induction differentiation.

AIM To construct differentiation subtractive cDNA library of human leukemia HL 60 cells induced by diallyl disfuld using suppression subtractive hybridization (SSH) technique and to clone differentiation associated genes in leukemia cells. METHODS Poly A + RNAs were isolated from HL 60 cells induced by dially disfuld and were reversely transcripted into double strand cDNAs (as tester). After the cDNAs were digested into short cDNAs with blunt ends, they were divided into two groups and were ligated to the special adaptor 1 and adaptor 2R, respectively. The tester cDNAs were then hybridized with driver cDNA from normal HL 60 cells and the products were amplified twice by nested PCR technique. The PCR products were ligated with pGEM T plasmid vectors and were transformed into E.coli JM109. The inserts of cDNAs were analyzed by restrictive enzyme EcoR I. RESULTS Subtractive cDNA library was constructed successfully and efficiently. Random analysis of 200 clones with enzyme restriction showed that about 84 5% clones contained 100～600 bp cDNA inserts. It can help clone novel genes associated with differentiation and explore the molecular mechanism of leukemia differentiation induced by diallyl disfuld. CONCLUSION DADS can induce human leukemia HL 60 cells differentiation, and suppression subtractive hybridization (SSH) technique is an effective method to isolate those differentially expressed genes.

Objective To explore the clinical significance of human papillomavirus L1 capsid protein detection in cervical exfoliated cells in high-risk HPV positive women. Methods From November 2012 to June 2013,386 high-risk HPV positive (detected by hybrid capture Ⅱ) cases were enrolled as eligible women from Huzhou Maternity&Child Care Hospital and Women′s Hospital,School of Medicine, Zhejiang University. All eligible women underwent liquid-based cytology (ThinPrep) followed by colposcopy. Biopsies were taken if indicated. Cervical exfoliated cells were collected for HPV L1 capsid protein detection by immunocytochemistry. Expression of HPV L1 capsid protein in groups with different histological diagnosis were compared, and the role of HPV L1 capsid protein detection in cervical exfoliated cells in cervical lesions screening was accessed. Results Total 386 enrolled eligible women were finally diagnosed histologically as follwed:162 normal cervix, 94 low-grade squamous intraepithelial lesion (LSIL), 128 high-grade squamous intraepithelial lesion (HSIL) and 2 squamous cervical cancer (SCC). The positive expression rate of HPV L1 in HSIL+(HSIL or worse) group was significantly lower than that in LSIL-(LSIL or better) group (19.2% vs 66.4%,P=0.000). While identifying HSIL+ in HPV positive cases and compared with cytology, HPV L1 detection resulted in significant higher sensitivity (80.77%vs 50.77%,P=0.000) and negative predictive value (NPV;87.18% vs 76.47%,P=0.004), significant lower specificity (66.41% vs 81.25%,P=0.000),and comparable positive predictive value (PPV;54.97% vs 57.89%, P=0.619). To identify HSIL+in HPV-positive/cytology-negative women, the sensitivity, specificity, PPV, and NPV of HPV L1 detection were 87.50%, 61.54%, 41.18%, and 94.12%respectively, while 80.00%, 86.36%, 80.00%and 86.36%respectively in HPV-positive/atypical squamous cell of undetermined significance(ASCUS)women. Conclusions HPV L1 capsid detection in cervical exfoliated cells have a role in cervical lesions screening in high-risk HPV positive women, and may be a promising triage for high-risk HPV-positive/cytology-negative or ASCUS women.

Aim To study the effects of cycle arrest and molecular mechanism in human leukemia HL-60 cells induced by diallyl disulfide ( DADS ) . Methods Cell count, colony formation in soft agar experiments and flow cytometry analysis were employed to observe the DADS-induced cell growth inhibition and the effect of cycle arrest in HL-60 cells. The expressions of Chk1/2 and its downstream element in HL-60 cells were detected by Western blot. Results Cell count revealed that population doubling time increased to 35. 03 h and 71. 82 h, respectively, from 19. 14 h in HL-60 cells treated with 60 and 120 μmol·L-1 DADS ( P<0. 05 ) . Colony formation in soft agar experiments showed that colony formation inhibition rate of HL-60 cells exposed to 30, 60, 90 and 120μmol·L-1 DADS increased to 35. 06%, 62. 10%, 93. 79% and 99. 35%, respectively ( P<0. 05 ) . Flow cytometry a-nalysis exhibited that HL-60 cells treated with 60 and 120 μmol · L-1 DADS for 24 h and 48 h arrested in G2/M phase in a concentration-and time-dependent manner ( P <0. 05 ) . Western blot disclosed that the expression of p-Chk1 increased in a time-dependent manner ( P <0. 05 ); however, Chk1, Chk2 and p-Chk2 were not changed in HL-60 cells treated with 60μmol·L-1 DADS (P >0. 05). The expression of Cdc25C, CyclinB1 and CDK1 decreased after treated with 60 μmol·L-1 DADS in a time-dependent manner ( P<0. 05 ) , but the expression of 14-3-3 protein did not change ( P>0. 05 ) . Conclusion DADS can in-hibit the proliferation of HL-60 cells, and induce G2/M arrest through Chk1/Cdc25 C/CyclinB1/CDK1 path-way.

Objective:To construct a replication-deficient adenovirus carrying p16 gene and to investigate its anti-tumor activity on human gastric cancer xenografts in nude mice.Methods:p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV,the latter was then used to recombine the replication-deficient adenovirus AdCMV-p16 in 293 cells.Human SGC-7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups:AdCMV-p16,Ad-LacZ,and control groups.Mice in AdCMV-p16 group received intratumoral injections of 2?108 pfu/100 l AdCMV-p16(injected every other day for 5 times).Mice in control group received the same volume of virus preserving solution.The tumor volumes were measured at predefined time points.The anti-tumor effect of AdCMV-p16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis.Results:The replication-deficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice(P0.05),only with a inhibition rate of 4.26%.The pathological examination showed that apoptoses were the main changes in AdCMV-p16 group,and p16 gene was found in the cancer cells.Conclusion:The replication-deficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.