Objective: To establish a method for the determination of ketoconazole and miconazole nitrate in compound ketocon-azole gels. Methods:An HPLC method was developed. A Hypersil BDS C18 column(416 mm × 200 mm,5μm) was used, the mobile phases consisted of 0. 5% ammonium acetate solution-methanol(containing 0. 2% triethanolamine) (20∶80), the flow rate was 1. 0 ml ·min-1 , the detection wavelength was set at 230nm, the column temperature was 30℃ and the injection volume was 20μl. Results:There was a good linear correlation within the range of 5. 1-510. 0 mg·L-1 for ketoconazole (r=0. 999 9) and 50. 0-500. 0 mg·L-1 for miconazole nitrate (r=0. 999 9), the average recovery for ketoconazole and miconazole nitrate was 100. 3%(RSD=0. 38%, n=6) and 99. 9%(RSD=0. 79%, n=6), respectively. Conclusion:The method is simple, rapid, accurate and sensitive, and can pro-vide a method for controlling the quality of compound ketoconazole gels.

Purpose To describe the clinical and pathological features of atypical vascular lesions (AVLs) after conservative surgery and radiation of the breast cancer,and to discuss the association with post-radiation angiosarcoma.Methods The clinical and pathological features in one case of AVLs was evaluated.The literatures were reviewed.Results The patient is a 57-yeAR~-old female who underwent a conservative surgery of the right breast because of a carcinoma.She received standard dose of radiation as adjuvant therapy.Three years later, multiple erythematous plaques developed around the former scar, which radiated to the nipple.Clinically,the plaques were considered as relapses of the carcinoma.However, fine needle aspiration gave negative results.Biopsy of one large plague revealed a circumscribed vascular lesion confined to the superficial dermis.It was composed of thin-walled anastomosing lymphatic vessels lined by attenuated endothelial cells.In focal areas, the vessels extended to the mid-dermis.Immunohistochemically, the endothelial cells were positive for CD31, CD34 and D2-40,with absent of α-SMA positive pericytes.Review of the breast tumor sections showed an invasive micropapillary carcinoma.Conclusions AVLs is a rare vascular lesion related with conservative surgery and post-radiation therapy of the breast.AVLs may represent as a precursor of breast angiosarcoma.Being familiar with the clinicopathologic characteristics of AVLs is important not only for the pathologists but also for the clinicians.

Objective To explore the effectiveness of sandplay therapy combined with family counseling in children with school phobia and its influence of child′ self-consciousness. Methods Integrative sandplay therary with family consulting were used to treat 28 patients with school phobia regularly for 2 months. Sandplay and family consulting therapy were given once a week for 45 minutes . Clinical outcomes were assessed using CGI-GI and Piers-Harris children′s self-consciousness scale before and after treatment as well as 3 months posttreatment. Results Overall response rate was 85%. In addition, the physical appearance and characteristic factor before and after treatment were no significant difference (P>0.05). The rest of the various factors and total score compared with pre-treatment significantly improved (P<0.05). After treatment for 3 months, every factor in self-consciousness of children and total score were no significant difference (P>0.05). Conclusion Integrative sandplay therapy with family counseling has better and long-lasting treatment effect to self-consciousness of children with school refusal.

BACKGROUND:A few studies have reported that autologous chondrocyte implantation is better than microfracture for treating cartilage defects of the knee. But there are few meta-analyses on the clinical outcomes of autologous chondrocyte implantation versus microfracture. OBJECTIVE:To evaluate the effects of autologous chondrocyte implantation versus microfracture in the treatment of cartilage defects of the knee based on existing clinical data. METHODS:A systematic search for control ed clinical trials or control ed prospective observational studies published from 1979 to January 2015 was done in electronic databases MEDLINE, EMBASE, CINAHL, the Cochrane Central Register, Wanfang, CNKI and VIP. The literatures about the effects of autologous chondrocyte implantation versus microfracture in the treatment of cartilage defects of the knee were retrieved. We screened the retrieved literature according to the inclusion and exclusion criteria and performed a Meta-analysis with the software RevMan 5.2 after identification of the relevant data. RESULTS AND CONCLUSION:Eight studies were enrol ed according to the selection criteria, which revealed a statistical y significant difference, representing a clinical y relevant superiority of autologous chondrocyte implantation over microfracture, in IKDC scores at final fol ow-up [weighted mean difference (WMD),-9.93;95%confidence interval (CI):-13.16 to-5.43;P<0.000 01] and available scores at 5-year fol ow-up [standard mean difference (SMD),-0.30;95%CI: -0.55 to-0.05;P=0.02). In contrast, there were no significant differences, thus representing no clinical relevant superiority of microfracture versus autologous chondrocyte implantation, in Tegner scores at final fol ow-up (WMD=0.44;95%CI:0.04 to 0.84;P=0.03), Lysholm scores at final fol ow-up (WMD=-10.21;95%CI:-33.68 to 13.26;P=0.39), and available scores at 2-year fol ow-up (SMD=-0.25;95%CI:-0.92 to 0.43;P=0.47). These findings demonstrate that autologous chondrocyte implantation can result in a better long-term outcome than microfracgure. However, whether autologous chondrocyte implantation has a better treatment effect than microfracture in general needs more research.

BACKGROUND:The bioceramics has the ideal pore size, high porosity and the through-hole rate, can provide the ideal physiological activity space for the bone cel repair, and can obviously improve bone conduction. OBJECTIVE:To explore the bone conduction and bone induction in the repair of bone defects in the stage of bone defect of bionic biphasic ceramic bioactive bone. METHODS:A total of 20 New Zealand white rabbits were randomly divided into bioactive glass and biomimetic biphasic ceramic bioactive bone groups, and were used to construct the animal bone damage model. They were given the repair with bioactive glass and biomimetic biphasic ceramic bioactive bone. RESULTS AND CONCLUSION:At 4 weeks after model establishment, scanning electron microscopy demonstrated that dense periosteal tissue was observed in the biomimetic biphasic ceramic bioactive bone group. At 8 weeks, dense combination was found, and no obvious fissure existed. At 12 weeks, complete bone demarcation blurred, showing a natural transition. Moreover, the binding site was very dense. There were a large number of new bone tissues, bone trabecula was regular and connected to a piece. The bone material has been largely degraded. Bone defects were repaired completely. The bone density was close to normal bone. At 8 weeks, in the bioactive glass group, the binding site presented obvious fissure. At 12 weeks, the fissure had been connected, but the binding was not tight as compared with the bionic biphasic ceramic biologic active bone group. The bone defect got preliminary repair. A smal number of new bone formed trabecular bone, but could not connect or traverse. There was no recanalization of the marrow cavity. A few continuous bone cal us traversed the broken end. These data demonstrate that bionic biphasic ceramic bioactive bone has good bone conduction, bone induction and biocompatibility in the repair of segmental bone defects.

Objective:To prepare the production of TIGIT-Fc fusion protein using H22 cells stably integrated the gene by lentivirus vector , and to explore the immunoregulatory effect on macrophages by TIGIT-Fc.Methods: TIGIT-Fc fusion gene were constructed by molecular cloning.The fusion gene was then subcloned to plasmids contained the secretion signaling peptide .The secrected TIGIT-Fc fusion gene was inserted into the lentivirus backbone vector.The purified lentivirus vector was the used to infect the murine H22 cell line.TIGIT-Fc protein was purified by protein A column from the ascites of H 22-injected C57BL/6 mice.Macrophages stimulated by lipopolysaccharide ( LPS ) was challenged to TIGIT-Fc treatment or control.Cytokine levels was then detected by ELISA.Results: TIGIT-Fc protein was purified from the ascites of H 22-injected mice.PVR was upregulated in LPS-treated macrophages.IL-10 level was upregulated in TIGIT-Fc treated macrophages.Conclusion: TIGIT-Fc promotes the mature macrophages to secrete anti-inflammatory cytokine IL-10.

Objective To study the chemical constituents from the exocarp of Juglans mandshurica.Methods The compounds were isolated by Diaion HP-20,Sephadex LH-20,MCI CHP-20,and silica gel column chromatography.Their structures were identified by physicochemical properties and spectroscopic analysis.Results Twelve compounds were elucidated as pinostrobin (Ⅰ),quercetin (Ⅱ),quercetin-3-O-?-D-glucoside (Ⅲ),chrysophanol (Ⅳ),gallic acid (Ⅴ),4-hydroxyl-cinnamic acid methyl ester (Ⅵ),vanillin (Ⅶ),caffeic acid (Ⅷ),4-hydroxylcinnamic acid (Ⅸ),?-sitosterol (Ⅹ),daucosterol (Ⅺ),and 5-hydroxyl-1,4-naphthoquinone (ⅩⅡ).Conclusion The compounds Ⅰ,Ⅳ,and Ⅵ—Ⅸ are isolated from J.mandshurica for the first time.

OBJECTIVE To explore the toxic effects of zinc oxide nanoparticls (ZnO-NPs) on cardiac development of zebrafish embryos and rat myocardial cell lines (H9c2),as well as potential molecular mechanisms.METHODS ZnO-NPs were characterized.Zebrafish embryos were exposed to different doses of ZnO-NPs (0,0.5,2.5,5.0 and 10.0 mg· L-1) for 24 to 96 h at 4 h post fertilization (4 hpf).The embryo mortality was observed.The expressions of notochord homeobox (noto),T-box 6 (tbx16),T,brachyury homolog a(ta),and tbx6 which were related to cardiac mesoderm were investigated using real-time PCR at 17 hpf.The heart rate and number of cardiomyocytes of embryos [[Tg (cmlc2:nucdsRed)] exposed to 0,2.5 and 5 mg· L-1 ZnO-NPs were detected at 72 hpf.Rat myocardial cell lines (H9c2) were treated with ZnO-NPs (0.1,0.5,1.0,5.0 and 10.0 mg· L-1) for 24 h.Cell viability was measured with Alamar Blue method.Mitochondrial ultrastructure was observed by transmission electron microscopy.Cellular ATP was detected using chemiluminescence,and oxygen consumption rate (OCR) was examined with Seahorse instrument.RESULTS The particle size of ZnO-NPs was (331 ±3)nm.The ZnO-NPs LC50 of zebrafish embryos at 48 hpf was 21.81 mg· L-1.The mRNA expressions of noto,ta and tbx6 were reduced after ZnO-NPs 2.5 mg· L-1 treatment at 17 hpf.The heart rate of 72 hpf zebrafish was 153 min-1 in the ZnO-NPs 5 mg· L-1 group,12.6％ lower than that in the cell vehicle group (P＜0.01),and the number of cardiomyocytes decreased by 15.5％ (P＜0.01) compared with the cell vehicle group.Reduced cell viability and mitochondrial vacuolation were observed in H9c2 after ZnO-NPs 0.5 mg· L-1 exposure.Compared with the cell vehicle group,the cell ATP decreased by 25.7％ (P＜0.05),and OCR decreased by 27.2％ (P＜0.01).CONCLUSION Low-dose ZnO-NPs exposure has effect on the cardiac development of zebrafish,mainly due to reduced heart rate and decreased number of cardiomyocytes.These changes may be related to the decreased expressions of cardiac development-related genes and the impairment of mitochondrial structure and function.

Objective　To investigate the feasibility of applying transvescal approach laparoendoscopic single-site radical prostatectomy (TVSSLRP) and assess the oncological and functional outcomes.Methods　Eight patients with clinically localized prostate cancer (PCa) of low risk underwent TVSSLRP.Demographic data were accrued including patient age,body mass index (BMI),preoperative PSA level,the International Index of Erectile Function 5,biopsy Gleason score,clinical TNM stage and D'Amico risk classification.One surgeon performed all TVSSLRP procedures.A homemade triple-port was introduced percutaneouly into the bladder to establish pneumovesicum through a 4 cm incision.The major steps of the surgery were described as follows:initial incision was made along posterior margin of the bladder neck to expose bilateral vas deference and spermatic vesicle.After opening Denonvilliers' fascia and extending the space to lateral prostatic pedicles,an intra-fascial nerve sparing procedure was performed.The puboprastatic ligaments were then separated close to the prostate surface and the dorsal vein complex was cautiously swept off.Subsequently,careful apical dissection and urethral transection was sequentially conducted. To reduce the tension of vesico-urethral anastomosis,3 additional incisions parallel to vesio-urethral margin were created and a novel tension - reduced V-LocTM barbed polydioxanone sutures was used.　Results　All the operations were successfully performed and there was no conversion to standard laparoscopic approach or open surgery.The total operative time range was 75 - 180 min with mean time of 125 min.The blood loss was 85 -450 ml with mean 140 ml and no blood transfusion was required.The catheter was removed after a mean (range) of 14 (9 -16) days.No intra-operative complications occurred. No patient had positive surgical margins.The mean (range) hospital stay was 17 (13 -25) days after surgery. All the cases were continent after removal of the catheter.No cases demonstrated vesico-urethral stricture and biochemical recurrence on 12 - 18 months follow up postoperatively.　Conclusions　TVSSLRP is technically feasible for cases with organ-confined prostate cancer with good oncological and functional results.

AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .