Objective:To observe the clinical treatment effect of Ginkgo mihuan oral liquid combined with tinnitus-curing instrument on the patients with sudden deafness associated with tinnitus.Methods: One hundred and twenty patients with sudden deafness associated with tinnitus were randomly divided into treatment group and control group one and control group two. Treatment group were treated with Ginkgo mihuan oral liquid combined with Tinnitus-curing instrument. The patients in control group were given Ginkgo mihuan oral liquid, the other control group used tinnitus-curing instrument. Observe the tinnitus and hearing improved progress.Results: In treatment group, the effective rate of tinnitus improvement was higher than that in control group, the differences were statistically significant(x2=7.040,x2=5.952;P<0.05), but the effective rate of hearing improvement was not higher than that in control group, the differences was not statistically significant(x2=0.621,x2=2.813;P<0.05).Conclusion: Ginkgo mihuan oral liquid combined with Tinnitus-curing instrument has certain curative effect on tinnitus recovery of patients with sudden hearing loss.

AIM:To compare the effects of alcohol and aqueous extracts from Huiyang shengji gao(HYSJG),a Chinese medicine,on proliferation of human fibroblasts derived from normal and ulcer margin skin.METHODS:Using explant culture techniques,we obtained populations of normal human fibroblast(nHFB)and ulcer marginal fibroblast(uHFB).Alarmar blue assay was used for observing cell proliferation,and flow cytometry for cell cycle analysis.RESULTS:After cultured with aqueous extracts and alcohol extracts of HYSJG for 24 hours,the proliferation rates of the two cell lines were both elevated.For alcohol extract,obvious proliferation-promoting concentrations on nHFB and uHFB were 0.027-0.425 mg/L and 0.027-0.213 mg/L,respectively(P

Chordoma ; pathology ; surgery ; Facial Neoplasms ; pathology ; surgery ; Female ; Humans ; Middle Aged ; Muscle, Skeletal ; transplantation ; Neoplasm Recurrence, Local ; surgery ; Reconstructive Surgical Procedures ; Surgical Flaps

Objective: To evaluate the effect of synthesized small interfering RNA targeting to HIF-lα on the adhesion and invasion of human tongue squamous cell carcinoma cell line (Tca8113). Methods; A double strand small interference RNA (siRNA) targeting HIF-1α (siRNAH1Fla) was transfected into cultured Tca8113 cells by lipofectamine2000. The expression of HIF-1α was investigated on mRNA level by real time-PCR and protein level by Western blot. The adhesion and invasion of Tca8113 cells to extracellular matrix (ECM) was also analyzed. Results: Exposure to hypoxia induced a prolonged elevation of HIF-lα protein and siRNAHIF.la reduced HIF-la synthesis as measured on mRNA level and protein level compared with the controls. No matter under normoxic or hy-poxic conditions, the adhesion potency of siRNAHIF-1α treated Tca8113 cells was markedly inhibited compared with controls(P<0.05 or P <0.01). So did the invasion potency (P<0.01). The adhesion and invasion potency of siRNAHIF.,a treated Tca8113 cells were inhibited more greatly under hypoxic condition than under normoxic condition ((36.4±2.7)% vs(26±2.35);(44.2±2.2)% vs (35±1.75), P<0.01)). Conclusion; siRNAH1F.lo can knockdown the expression of HIF-la and inhibit the cell adhesion and invasion to ECM in Tca8113 cells. HIF-la may play an established role in the regulation of Tca8113 cells invasion and metastasis. Interfering with HIF-1α pathways by siRNA strategy may provide a therapeutic target for human tongue squamous cell carcinomas.

Objective To investigate relationship of platelet (PLT) count with clinicopathological features and prognosis of patients with breast cancer,and explore the susceptibility index to evaluate prognosis of patients with breast cancer.Methods A retrospective analysis of clinical data of 498 patients with breast cancer in January 1995 to December 2005 was carried out.PLT count was tested.Those patients were divided into group A (PLT ＜ 150 × 109/L),group B[(150-250) × 109/L],and group C (PLT ＞ 250 × 109/L) according to PLT count level.The relationship of platelet count with clinicopathological features was analyzed.Kaplan-Meier and Cox proportional hazards model were used for univariate analysis and multivariate analysis of PLT impact on survival time.Results There was positively correlated between PLT count and clinicopathological features (Pearson coefficient ＞ 0,P ＜ 0.05).There was negative correlated between PLT count and survival time (Pearson coefficient =-O.583,P ＜ 0.05).The survival time of groups A,B and C were significantly different (P =0.018).Cox proportional hazards model multi-factor analysis showed that PLT count was an independent factors affecting survival time (OR =2.256,P ＜ 0.05).Conclusions Patients with breast cancer associated with increased emphasis and PLT count.PLT count had negative correlation with survival time.PLT count could be a susceptible index to predict the prognosis of patients with breast cancer.

Objective:To prepare rabbit polyclonal antibody against human argonaute 3(AGO3) protein,to identify its properties and investigate the tissue distribution of AGO3 using tissue array.Methods:AGO3 peptide was synthesized using chemical method,and then conjugated to Keyhole limpet hemocyanin(KLH) as immunogen.The AGO3-KLH conjugate were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification using affinity chromatography.Then the distribution of AGO3 in tissues was examined through immuno-stainning by tissue array.Results:Rabbit polyclonal antibodies against AGO3 were raised after immunization with AGO3-KLH conjugates.The anti-serum titer after the last inoculation was up to 1∶20 000.The preparations of the antibody were confirmed to raised recognize AGO3 peptides specially by ELISA and Western blot.AGO3 protein was stained positively in the cytoplasm of tumor cells and epithelial cells in many normal tissues.Conclusion:The polyclonal antibody against AGO3 protein has been achieved successfully,and it provides an efficient tool for further studying the roles of AGO3 in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.

Objective To prepare polyclonal antibodies against human PIWIL and to identify their property and tissue distribution of PIWIL.Methods PIWIL polypeptide was synthesized and conjugated to Keyhole limpet hemocyanin(KLH) as an immunogen.Then PIWIL-KLH conjugations were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification by affinity chromatography.PIWIL were then stained on the tissue chip to study their distribution.Results Rabbit antibodies against PIWIL were prepared after injection of PIWIL-KLH conjugation.These antibodies specially recognized PIWIL peptides.Expression of PIWIL was found in the cytoplasm of epithelia cells of varied normal tissues and tumor tissues.Conclusion The successful preparation of the polyclonal antibody against PIWIL will provide an efficient reagent for further study of its role in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.

Sailuotong (SLT) is a compound preparation composed of ginseng, ginkgo and saffron for the treatment of vascular dementia In order to identify its material foundation and provide evidence for therapeutic regimen, the pharmacokinetics and brain distribution of ginsenosides were investigated after intragastric administration of SLT. An LC-MS/MS method was developed for the determination of 7 ginsenosides in rat plasma simultaneously. Statistical analysis of obtained data demonstrated that the method has achieved the desired linearity, precision, accuracy and sensitivity. After administration of SLT at 60 mg x kg(-1) dose, 7 ginsengosides were all absorbed into systematic circulation. The quantitative and statistical analysis of gensenosides in plasma showed that protopanaxdiol saponins exhibited higher concentration and longer half life than protopanaxatriol saponins. The mean value of half life of ginsenosides Rg1, Re, Rb1, Rb2/b3, Rc and Rd were 15.26, 2.46, 18.41, 27.70, 21.86 and 61.58 h respectively. The peak concentration of them were 7.15, 2.83, 55.32, 30.22, 21.42, 8.81 microg x L(-1) respectively. The determination of brain distribution at different time after dosing revealed ginsenosides entered into brain promptly but the concentration declined along with time rapidly. The ginsenosides with higher concentration in brain were Rg1, Re, Rb1 and Rc. These findings demonstrated ginsenosides could be absorbed in blood and penetrated into brain rapidly. Some ginsenosides, especially Rg1 and Re, might be the main components directly effecting neurocyte in brain taking advantage of their better brain distribution. While ginsenosides of mostly protopanaxdiol saponins might protect brain mainly depending on peripheral efficacy in virtue of their long residence in blood, by which higher concentration could be reached after multiple dosing.
Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Ginsenosides ; pharmacokinetics ; Male ; Rats ; Rats, Wistar ; Time Factors

AIM: To investigate the effects of Astragalus polysacharin(APS) on human fibroblast and human umbilical vein endothelia cell (HUVEC) proliferation, as well as its acts on adhesion between white cells and HUVECs. METHODS: Human fibroblasts from distal and proximal skin away the ulcer were cultured as normal fibroblasts(NF) and wounded fibroblasts(WF). MTT assay was used for detecting cell proliferation, Rose Bengal staining and fluorescence immunohistology assay were used for examining the adhesion of human polymorpho-nuclear cell(PMN) and TPH-1 to HUVECs. RESULTS: 2 44-156 mg/L APS promoted WF proliferation, and 2 44-39 mg/L APS also promoted NF proliferation, but it did not show any proliferating effect on HUVECs. APS inhibited the adhesion of PMN or TPH-1 to HUVECs induced by tumor necrosis factor(TNF). At 25-100 mg/L, it also inhibited both VCAM-1 and ICAM-1 expression in HUVECs induced by TNF. Treatment with APS for 12 h also inhibited CD44 expression in HUVECs. CONCLUSION: APS shows mitogenic activity on both human normal and wounded fibroblasts. It also exerts anti-inflammation effects by inhibiting adhesion molecule expression and adhesion of white cells to HUVECs. [

Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P＜0.05).Furthermore,a notable decreased cell survival rate (38.3％) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8％ in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2％ and 10.9％ respectively,F=19.926,45.409;both P＜0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.