Objective To determinate Berberine chloride in compound Wuzhigan capsules and develop a steady and reliable method for the quality control of the capsules.Methods A HPLC method was adopted.The column was Diamonsil C18 (250 mm?4.6 mm,5 ?m) with the column temperature of 30 ℃,the mobile phase was acetonitrile-0.033 mol/L potassium dihydrogen phosphate (30∶70) with the flow rate of 1.0 mL/min,and the detection wavelength was at 347 nm.Results Berberine chloride has a linearity in the range of 2.52～12.6 ?g/mL,and the average content in the capsules was 1.686 mg/capsules.Conclusion This method is sample,accurate,and can be used for the quality control of compound Wuzhigan capsules.

Purpose To find the reasons and explore the appropriate countermeasures for failure ureteroscopy operation.Methods The clinical data of 74 cases with failure ureteroscopy operation for ureteral calculi were analysed respectively.Results The failure reasons including the difficult of ureteroscopy insert (46 lateral),difficult of calculi disposal (12 lateral) and ureteral injure (15 lateral).ESWL(34 sides) and open surgery(44 sides) were remedial measures after failure ureteroscopy and the success rates were 85 3%, 100% respectively.nephrectomy were carried out in and 1 case died from serious uriaemia.The mortality was only 1.4%.Conclusions The reasons of failure including unable approach the stone, difficult to disposal calculi and ureteral injury. The effect is related to applicances, experience, indications . ESWL and open surgery are the suitable remedy measures according to the failure reason.

Objective To evaluate the clinical value of transrectal ultrasound guided systematic 13 cores prostate biopsy. Methods A total of 160 patients referred for abnormal digital rectal examination and (or) prostate specific antigen 4ng/ml or greater underwent transrectal ultrasound guided systematic 13 cores prostate biopsy.That was,in addition to the standard sextant biopsies,cores were taken from the far lateral and middle regions of the gland as described by Eskew.Pathological findings of the additional regions were compared with those of the sextant regions. Results Of the patients 35 % had cancer on biopsy(56 /160).Of the 56 patients with prostate cancer 12(21%)had carcinoma only in the additional regions,which would be undetected on the sextant biopsy technique (P

Objective To evaluate the expression and discussion on high mobility group protein ( HMGB1 )/toll like receptor 4 ( TLR4 ) and NF-κB possible role in the signaling pathway in preeclampsia .Methods Ten patients with mild preeclampsia(MP), 20 patients with severe preeclampsia (SP) and 30 cases of normal pregnancy were recruited the same period.To check the expression of the HMGB1,TLR4,NF-κB P65 protein in placenta tissue using Immunohistochemical staining .Levels of HMGB1, TLR4, NF-κB P65 in blood serum were measured by ELISA.Results 1)ExpressionofHMGB1,TLR4,NF-κBP65wereincreasedascomparetocontrolgroup(P<0.05);HMGB1,TLR4,NF-κB P65 in placenta of patients with severe preeclampsia and mild preeclampsia failed to show significant difference .2) HMGB1,TLR4,NF-κB P65 in women with preeclampsia was significantly higher than control group ( P<0.05 ); HMGB1,TLR4,NF-κB P65 in women with severe preeclampsia showed a higher level as compared to mild preeclampsia (P<0.05).Conclusions HMGB1,TLR4 and NF-κB P65 were over-ex-pressed in the placenta tissue and serum in patients with preeclampsia , which indicated that HMGB1,TLR4 and NF-κB P65 may play an important role in the development of preeclampsia .

Objective To evaluate the alteration of CD4+ regulatory T cells in peripheral blood from patients with ankylosing spondylitis(AS). Methods Seventy-eight AS patients and 50 healthy individuals were included in this study. The proportion of CD4+CD25+CD127lo/- T cell population in CD4+ T cells as well as that cytotoxic T lymphocytes(CTL) and NK cells in lymphocytes was evaluated by flow cytometry. Serum TGF-β and TNF-α were measured by ELISA. The inhibitory function of CD4+CD25+ regulatory T cells was measured by mixed lymphocyte culture. Results The population of CD4+CD25+CD127lo/- T cells in peripheral blood of AS patients accounted for (4.36±1.21)% of CD4+ T lymphocytes, which was significantly lower than that in healthy individuals (P<0.05). The CD4+CD25+CD127lo/- T cells population in AS patients was positively correlated with TGF-β level, but negatively with TNF-α. Compared with healthy individuals, the function of CD4+CD25+ regulatory T cells in the inhibition of alloreactive T cells was lower in AS patients, which was related to the decreased secretion of TGF-β. Conclusion The CD4+ regulatory T cells in peripheral blood of AS patients are significantly decreased and its function is defective, which leads to immune regulatory dysfunction in vivo. It may be one of immune pathogenesis mechanisms of AS.

Objective To analyze the clonal gene rearrangement and complementarity determining region 3 (CDR3) repertoire of TCR β-chain in fine needle aspiration biopsy (FNAB) specimens of lymph node metastasis in patients with breast cancer. Methods The TCR CDR3 region genes of 24 TCR Vβ subfamilies were amplified by utilizing RT-PCR technology, and the CDR3 lengths of TCR β-chain were analyzed with gene scan technology for 2 cases with lymph node reactive hyperplasia and 3 patients with lymph node metastasis of breast cancer. The clonality of T cells presumed by spectra typing was further confirmed by CDR3 sequencing. Results TCR β-chain presented specific repertoire skewing in metastatic lymph node,and only 3-5 TCR Vβ subfamily of T cells were identified, respectively. Clonal expanded T cells, including oligoclonal, polyclonal patterns, in one or more Vβ subfamilies were found in all cases. The oligoclonal expanded T cells had different CDR3 amino acid sequences. Conclusion There are characteristic T cells cloning proliferation and selected usage of TCR Vβ subfamily T cells could be found in metastatic lymph node.The sequences of CDR3 in different TCR clone proliferation are mostly different.

Objective To investigate the effects of ghrelin on proliferation of vascular smooth muscle cells(VSMC)and the expression of mitochondrial fusion 2(Mfn-2)in cultured human aortic smooth muscle cells(HASMCs).Methods HASMCs were cultured in vitro,treated with different concentrations(10-9,10-8,10-7,10-6,10-5 mol/L)ghrelin or 10-6 mol/L ghrelin for different time(0,6,12,18,24 h).Subconfluent HASMCs at passage 4-6 were used in experiments.MTT essay was used to investigate the effect on proliferation of HASMCs.RT-PCR and Western blot were used to analyse the expression of Mfn-2.Results 10-7-10-5 mol/L ghrelin inhibited the proliferation of HASMCs,and the inhibitory effect of concentration of 10-6 mol/L was the most obvious(P<0.01).Ghrelin inhibited the proliferation of HASMCs in 6-24 h,and it reached the peak at 24 h(P<0.01).10-6 mol/L ghrelin significantly increased the expression of Mfn-2 mRNA and protein(P<0.01).The up-regulation of 10-6 mol/L ghrelin on Mfn-2 mRNA and protein expression reached the peak at 18 h(P<0.01).Conclusion Ghrelin might inhibit the proliferation of HASMC by up-regulating the expression of Mfn-2.

AIM To evaluate whether l glutamine monofluorophosphate(MFP) together with Alendronate sodium may further increase bone mass in ovariectomized rats. METHODS Forty two 3 month old Sprague Dawley female rats were randomized into six groups:group 1 rats were sham operated(Sham),group 2 rats were ovariectomized controls(Ovx),and groups 3～6 were ovariectomized and received either 1 mg?kg -1 ?d -1 of alendronate sodium ,270 mg?kg -1 ?d -1 of calcium gluconate,5 6 mg?kg -1 ?d -1 of MFP or combination of 5 6 mg?kg -1 ?d -1 of MFP and 1 mg?kg -1 ?d -1 of alendronate sodium for 3 months. All animals received double bone fluorochrome labeling prior to sacrifice. At the end of experiment,the left tibiae were havested for histomorphometrical evaluations. RESULTS Alendronate sodium increased trabecular bone volume significantly with suppressed bone resorption and bone formation. Administrated calcium gluconate had no influence on the bone mass. MFP also could not restore cancellous bone of ovariectomized rats. Administration of both MFP and alendronate sodium increased bone mass significantly but did not increase bone mass compared with alendronate sodium. CONCLUSIONS The results indicated administration of both alendronate sodium and MFP could not further increase bone mass of ovariectomized rats.

Objective To investigate the clinical effect of ropivacaine hydrochloride at 0.25%, 0.375% and 0.5% concentration for lower limb nerve block anesthesia.Methods75 cases of lower extremity nerve block anesthesia from Ningbo Zhenhai District Hospital of traditional Chinese Medicine from September 2014 to February 2015 were enrolled in the course of the study, they were divided into three groups: group 0.25%, the equivalent number of 0.375% and 0.5% groups, and three groups of patients were made with a concentration of 0.25%, 0.375%, 0.5% ropivacaine hydrochloride for lower extremity nerve block anesthesia;the clinical data of three groups were analyzed retrospectively, observation of three groups of patients with lower limb nerve block effect.ResultsThe results showed that 0.25% groups of patients in the motor block time was (36.8±5.9) minutes, motor block in patients with a total of 7 cases, with nerve block in patients with a total of 11 cases.0.375% groups of patients in the motor block time was (23.1±4.3) minutes, motor block in patients with a total of 14 cases, with nerve block in patients with a total of 18 cases.0.5% groups of patients in the motor block time was (20.6±5.7) minutes, motor block in patients with a total of 16 cases, with nerve block in patients with a total of 18 cases.Available block time of 0.375% groups of more than 0.5% groups, less than 0.25% groups;0.375% motor block group was 56.0%, higher than the 0.25% group, lower than the 0.5% group;the 0.375% group and 0.5% group of nerve block ratio is same, higher than the 0.25% group, the difference was statistically significant (P<0.05).ConclusionThe effect of anesthesia using concentration of 0.375% ropivacaine hydrochloride is relatively good, can reduce the dosage of anesthetic drugs, but also ensure the anesthetic effect, meet the clinical requirement, this method will be applied to.

Purpose Liver fibrosis is the common pathological process of cirrhosis developed by chronic liver disease.This study aims to explore the application of microvascular change in assessing the liver fibrosis induced by bile duct ligation based on X-ray phasecontrast computed tomography (PCCT).Materials and Methods Ten rats were equally divided into the group of bile duct ligation-induced liver fibrosis and the control group at random.Then ten excised liver tissues were imaged at beamline BL13W1 of Shanghai Synchrotron Radiation Facility in China.After the imaging studies,the liver samples were stained with Sirius red and immunostained with antibodies against cytokeratin 19 (CK19).The microvasculature in livers was reconstructed via PCCT,and the correlation analysis on microvascular density and the proportion of fibrosis area was performed.Results The microvasculature including blood vessels,dilated bile ducts and ductular proliferation were clearly revealed by PCCT technology.Moreover,the 3D microvascular density and proportion of fibrosis area in the liver fibrosis group increased significantly compared with those in the control group (P＜0.01),and the correlation between them was obvious (r=0.812,P＜0.01).Conclusion Microvascular 3D characteristics of liver fibrosis are well demonstrated by PCCT,and 3D microvascular density is significantly correlated with the proportion of fibrosis area.Those findings indicate that microvascular change is closely related to the degree of liver fibrosis induced by bile duct ligation,and thus it provides a novel means to assess the severity of liver fibrosis through microvascular change.