Objective To explore the mechanism of the formation of glioma cells derived vessels (GCDV).Methods The tubular formation assay was performed on 3D cell cultures of U87,U251,U373,SF295,T98G,SKMG-4 and C6 glioma cell lines.The expression of Notch1 、Dll4 were examined by western blot analysis.Result The mean number of vasculogenic channels of cell lines C6,U373,U87,T98G,SF295,U251 and SKMG-4 per area (100×) were 25.2±5.0,36.4±3.2,19.0±2.2,12.6±2.4,4±2.2,0.2±0.4 and 0,respectively.The relative protein densities of Notchl in U87,U251,T98G,SF295,SKMG-4,C6 and U373 were 0.34,0.21,0.79,0.04,0.28,1.75 and 1.19,which were significantly related with the tubular formation ability (P＜0.05).However,the expression of Dll4 was not associated with tubular formation ability (P＞0.05).Conclusion Notch1 may play a key role in tubular formation of glioma cells while the role of Dll4 in the process needs further study.

Objective To investigate the effect of the calmodulin kinase Ⅱ Inhibitor KN-93 on L-typecalcium current(ICa,L)and intracellular calcium concentration([Ca2+]i)in hypertrophic cardiac myocytes.Methods Forty-eight female New Zealand white rabbits were randomized(random number)into four groups(12 animals in each group):the sham operation group(sham group),the left ventricular hypertrophy group(LVH group),the myocardial hypertrophy + KN-93 group(KN-93 group),and the myocardial hypertrophy + KN-92 group(KN-92 group).Myocardial hypertrophy in the rabbits was established by coarctation of the abdominal aorta.In the sham group,the abdominal aorta was dissociated without coarctation.Eight weeks after coarctation,single ventricular myocytes were isolated by enzymaticdissociation,and ICa,L was recorded using perforated-patch recording(PPR)techniques.[Ca2+]i was measured using single-cell calcium imaging with the fluorescence calcium indicator dye fura-2/AM.Results Cardiac hypertrophy was successfully established after 8 weeks of coarctation of the abdominal aorta.The peak ICa,L in the LVH group and the sham group was(1.38 ± 0.3)nA and(0.87 ± 0.1)nA at 0 mV,respectively(P ＜0.01,n =12).There was no significant difference in Ica,L density between the LVH group and the sham group[(6.7 ±1.0)pA/pF vs.(6.3±0.7)pA/pF; P≥0.05,n=12].The addition of either KN-92(0.5 μmol/L)or KN-93(0.5 μmol/L)to the perfusing solution caused a modest steady-state inhibition of peak ICaL(9.4％ ±2.8％,KN-92; 10.5％ ±3％,KN-93)(P≥0.05,n =12)at 0 mV.However,at a higher concentration(1 μmol/L),KN-93 more potently inhibited peak ICa,L(40％±4.9％)compared to KN-92(13.4％ ± 3.7％ ; P ＜ 0.01,n =12).Resting[Ca2+]i levels in fura-2-loaded myocytes isolated from the sham,LVH,KN-92,and KN-93 groups were(98 ± 12.3)nmol/L,(154 ± 26.2)nmol/L,(147 ± 29.6)nmol/L,and(108 ± 21.2)nmol/L,respectively.Conclusions The CaMK Ⅱ specific inhibitor,KN-93,can effectively block ICa,L and reduce intracellular calcium overload in hypertrophic cardiac myocytes.This action may account for the antiarrhythmic effect of KN-93 in hypertrophic ventricular myocardium.

Objective To investigate the Human papillomavirus(HPV)infection in different age groups of women in Shenyang, and explore its correlation with cervical biopsy diagnosis.Methods 7 311 women aged 13-85 did HPV test and thin-cytologic test (TCT)in the hospital.Some of them had biopsy detection under electronic colposcopy,and the pathological diagnosis was the golden standard for the diagnosis of cervical lesions.SPSS18.0 statistical software was used for all statistical analysis.Results The infection rate of <30 years old women was significantly higher than that of 30 - <40,40 - <50,≥50 years old women (P <0.05).The most prevalent high-risk HPV genotype in Shenyang were subtype 16,52,58,53,33,31 and 18,and the most prevalent low-risk HPV subtypes were 81,11 and 6.The former 4 subtypes of high-risk HPV infection accounted for 67.3% of all high-risk infection.As to the 4 subtypes with higher infection rate,the infection rate of ≥40 years old women was higher than that of <40 years old(χ2 =20.29,P =0.00).The top two low-risk HPV subtypes accounted for 74.8% of the infections.The mean age of the ICC patients were 48.3,which was statistically different from the other groups(P <0.05).Cervical lesions occured mostly in 40-49 years old,which accounted for 37.1% and was higher than the other agees(P <0.01).HPV16 infection rate increased with the severity of cervical lesions.Conclusion HPV DNA genotyping is a necessary methord for cervical cancer screen,an effective com-plement for precancerous lesions diagnosis which was missed in cytology test,and also an indispensable test for CIN treatment and follow-up after operation.

BACKGROUND:n mandibular posterior dental implantation,injury to the inferior alveolar nerve sometimes occurs because of mandibular canal going across mandibular body.This restricts the use of dental implantation at this site.Therefore,it is essential to understand the anatomic structure of inferior alveolar nerve canal in mandibular posterior dental implantation.OBJECTIVE:To observe the intramandibular course of and anatomic structure of inferior alveolar nerve canal.METHODS:Fifteen adult complete mandible specimens with teeth and 4 fresh mandible arterial infusion specimens were researched.All the specimens had complete dentition and there were no obvious absorption in alveolar bone.The course of inferior alveolar nerve canal and its dimension including transverse and longitudinal diameters of mandibular canal and the distance between mandibular canal and mandible each side (superior,inferior,buccal and lingual side) were measured in 15 adult mandibles with teeth.The relationship between blood vessels and nerve of the canal was observed in 4 fresh arterial infusion specimens.RESULTS AND CONCLUSION:The distance between the medial border of the mandibular canal and the lingual wall was shorter than that of the lateral wall of the mandibular canal to the buccal wall (P < 0.01);The length from the upper wall of mandibiular canal to the top of the alveolar ridge was longer than that of the inferior border of the mandibular canal to the inferior border of the mandible (P < 0.01).The longitudinal diameter was smaller than the transverse diameter (P < 0.05),namely,the cross section of the mandibular canal was an ellipse with a longer longitudinal diameter.There was no significant difference between the transverse and longitudinal diameters of the canal in the anterior and posterior teeth region of the mandible.The inferior alveolar nerve and its associated blood vessels were located within a nervous vascular bunch in the mandibular canals.In every fresh specimen the blood vessels lay above the nerve.There were small branches of blood vessels surrounding thenerve.The mandibular canal ran towards the lingual side and was close to the inferior margin of the mandible.

(3S)-Linalool synthase (LIS) is a key enzyme involved in the monoterpene biosynthetic pathway. Based on our previous transcriptome study, the expression level of LIS gene was exceedingly related to glycyrrhizic acid (GA) biosynthesis. Therefore, we used hairy root culturing to further investigate the effect of LIS on the GA biosynthesis. A LIS gene (GenBank accession number: MZ169552) was cloned from Glycyrrhiza uralensis. The plant binary overexpression vector pCA-LIS was constructed by gene fusion. G. uralensis hairy roots overexpressing LIS were induced by the Agrobacterium rhizogenes ATCC15834. The expression levels of LIS were analyzed by real-time quantitative PCR (RT-qPCR) and the contents of GA in hairy root lines were determined by UPLC. It was found that in the hairy root lines overexpressing LIS, the expression levels of LIS were significantly higher than that in the wild type, while the contents of GA were remarkably lower than those in the wild type and negative control. These findings indicate that the expression level of LIS is negatively correlated with the accumulation of GA. In this study, LIS was cloned from G. uralensis for the first time and the negative regulatory effect of LIS on GA biosynthesis was confirmed by reverse genetics. This work provides support for further improvement of the molecular regulatory network of GA biosynthesis in G. uralensis.

Ginkgo is one of the most successful cases of botanical drugs developed by modern science and technology during the past fifty years all over the world. At present ginkgo has been applied to the prevention and treatment of cardiovascular disease widely, and has good clinical efficacy. Type 2 diabetes has been proved to be the risk equivalents of cardiovascular disease, therefore it has an important scientific significance for looking for more effective drugs of prevention and control of diabetes. To seek more efficient and safe drug from the plant medicine which has the function of regulate blood sugar and improve insulin resistance becomes a hotspot at home and abroad. Basic and clinical studies have shown the ginkgo preparations of Chinese medicine have certain regulation effect on blood sugar and insulin resistance. In this paper, we review the mechanisms and clinical applications of ginkgo preparations on diabetes and its applications during the past 10 years.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Ginkgo biloba ; chemistry ; Humans ; Hypoglycemic Agents ; administration & dosage

Objective To investigate the expression of integrin α6 mRNA in skin lesions of patients with condyloma acuminata. Methods Sixty outpatients with condyloma acuminate, including 30 primary cases and 30 recurrent cases were enrolled in the study;30 non-condyloma acuminate patients undergoing circumcision were used as normal controls. The expression of integrin α6 mRNA in condyloma acuminata lesions of primary and recurrent cases, as well as in skin samples of normal controls were determined by RTPCR. ANOVA was used to compare the differences among the groups, and Student-Newman-Keuls test(q test)was used for pairwise comparisons. Results Relative contents of integrin α6 mRNA(integrin α6/GAPDH)in normal controls, primary and recurrent condyloma acuminata lesions were 0.25±0.10, 0.79±0.16 and 1.07±0.29, respectively, and the difference was of statistical significance(F=127.687, P=0.001). Conclusion Integrin α6 may be associated with the pathogenesis and recurrence of condyloma acuminate, which may provide a new target for prevention and treatment of the disease.

Objective: To examine expression patterns of focal adhesion kinase (FAK) and its activated form, phosphorylated FAK (pFAK),in human osteosarcoma and to investigate the correlation of FAK expression with clinicopathological parameters and prognosis. Functional consequence of manipulating FAK protein levels was also investigated in human osteosarcoma cell lines. Methods: Immunohistochemical staining was used to detect FAK and pFAK levels in pathologically archived materials from 113 patients with primary osteosarcoma. Kaplan-Meier survival and Cox regression analyses were used to evaluate prognoses. The role of FAK in cytological behavior of MG63 and 143B human osteosarcoma cell lines was studied via the FAK protein knockdown with siRNA. Cell proliferation, migration, invasiveness, and apoptosis were assessed using cell counting kit-8, Transwell, and Annexin V/PI staining methods. Results: Both FAK and pFAK were overexpressed in osteosarcoma patients. Tumor cells exhibited cytoplasmicity and occasional membranous immunoreactivity for FAK. A total of 42 cases (37.17%) mainly showed expressed pFAK in cytoplasm of osteosarcoma cells. No overexpression staining of anti-FAK and anti-pFAK antibodies was observed in normal cancellous bone tissues or negative controls. Significant differences were observed in overall survival between FAK-/pFAK- and FAK+/pFAK- groups (P=0.016), FAK+/pFAK- and FAK+/pFAK+ groups (P=0.012), and FAK-/pFAK- and FAK+/pFAK+ groups (P<0.001). All groups showed similar metastasis-free survival. Cox proportional hazard analysis showed that FAK expression profile is an independent indicator of both overall andmetastasis-free survival. siRNA-based knockdown of FAK significantly reducedmigration and invasion of MG63 and 143B cells and affected proliferation and apoptosis in osteosarcoma cells. Conclusion: Osteosarcoma malignancies in vitro and in vivo were correlated with overexpression and phosphorylation of FAK. These findings suggest that FAK plays an important biological role in osteosarcoma carcinogenesis. This study provides a better understanding of diagnostic and prognostic relevance of FAK overexpression and phosphorylation in osteosarcoma patients. Therefore, FAK and pFAK can be used as independent predictors of overall and metastasis-free survival in osteosarcoma patients.

Vasculogenic mimicry (VM) is a process by which aggressive tumor cells generate non-endothelial cell-lined channels in malignant tumors including hepatocellular carcinoma (HCC).It has provided new insights into tumor behavior and has surfaced as a potential target for drug therapy.The molecular events underlying the process of VM formation are still poorly understood.In this study,we attempted to elucidate the relationship between Notch4 and VM formation in HCC.An effective siRNA lentiviral vector targeting Notch4 was constructed and transfected into Be17402,a HCC cell line.VM networks were observed with a microscope in a 3 dimensional cell culture system.Cell migration and invasion were evaluated using wound healing and transwell assays.Matrix metalloproteinases (MMPs) activity was detected by gelatin zymography.Furthermore,the role of Notch4 inhibition in Be17402 cells in vivo was examined in subcutaneous xenograft tumor model of mice.The results showed that downregulation of Notch4 destroyed VM network formation and inhibited migration and invasion of tumor cells in vitro (P＜0.05).In vivo,tumor growth was also inhibited in subcutaneous xenograft model (P＜0.05).The potential mechanisms might be related with down-regulation of MT1-MMP,MMP-2,MMP-9 expression and inhibition of the activation of MMP2 and MMP9.These results indicated that Notch4 may play an important role in VM formation and tumor invasion in HCC.Related molecular pathways may be used as novel therapeutic targets for HCC antiangiogenesis therapy.

Objective To explore effect of endogeneous gangliosides(Gls) and integrin ?2?1 on protein phosphotyrosine expression of pp125 focal adhesion kinase (pp125FAK) after adhesion of SK-N-SH neuroblastoma cells to collagen(Col).Methods SK-N-SH cell line with high expression of integrin ?2?1 was cultured in presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol(D-PDMP).Effect of endogeneous Gls,anti-?2 and anti-?1 monoclonal antibody on protein phosphotyrosine expression of pp125FAK during adhesion of SK-N-SH cells to Col were determined by immunoprecipitate and Western blotting.Results After 6 days,endogenous Gls in cells were almost depleted.Gls-depletion,anti?2 and anti-?1 monoclonal antibody were able to decrease pp125FAK expression of SK-N-SH cells adherent to Col respectively.GD2,the major component of neuroblastoma cell Gls could reco-ver pp125FAK expression to a certain degree.Conclusions Endogenous tumor Gls regulate protein phosphotyrosine expression of pp125FAK during adhesion of neuroblastoma cells to Col.It is suggested that tumor Gls may increase signal transduction of tumor cell integrin ?2?1 by increasing tyrosine phosphorylation of pp125FAK.