Objective:To investigate the efficacy of Balanophora involucrata Hook.f.in treatment of hyperuricemia(HUA)based on network pharmacology,molecular docking,and hyperuricemia models in vivo and in vitro,and to clarify the main targets of its active components and related signaling pathway mechanism.Methods:The potential targets of Balanophora involucrata Hook.f.in treatment of HUA were identified by Databases such as the Traditional Chinese Medicine Database in Taiwan,the Chinese Herbal Medicine Identification Database,Professional Chemical Database,TargetNet Database,SwissTargetPrediction Database,GeneCards,Therapeutic Target Database(TTD),DrugBank Database,DisGeNET Database,Online Mendelian Inheritance in Man(OMIM)Database,and Venny Database.STRING Database and Cytoscape software were used to construct the active component-predictive target network and protein-protein interaction(PPI)network for Balanophora involucrata Hook.f.;topological analysis was used to select the main active components and core targets;Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed by R software;AutoDock Vina software was used for molecular docking validation.The NRK-52E cells were divided into blank control group,blank administration group,model group,and different concentrations(2.0,10.0,and 50.0 μmol·L-1)of erythrodiol(EDT)groups.High-performance liquid chromatography culture(HPLC)was used to detect the uric acid(UA)levels in the cell culture supernatants in various groups.The male ICR mice were divided into blank control group,blank administration group,model group,and EDT group;the mice in the last two groups were used to prepare the HUA models;kits were used to detect the levels of UA,creatinine(Cr),and blood urea nitrogen(BUN)in serum of the mice in various groups;the bilateral kidney tissue of the mice was harvested and weighed;the kidney indexes of the mice in various groups were calculated;TUNEL staining was used to observe the apoptosis in kidney tissue of the mice in various groups;Western blotting method was used to detect the expression levels of protein kinase B(AKT),phosphorylated AKT(p-AKT),phosphoinositide 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and matrix metalloproteinase-9(MMP-9)proteins in kidney tissue of the mice in various groups.Results:Six active components of Balanophora involucrata Hook.f.were identified,involving 116 intersecting targets and 14 core targets.The enrichment analysis yielded 1 828 GO terms and 145 signaling pathways.The molecular docking results showed that EDT had good binding activity with MMP-9.The high uric acid cell experiment results showed that compared with blank control group,the UA level in the cells in model group was significantly increased(P<0.01);compared with model group,the UA levels in the cells in 2.0,10.0,and 50.0 μmol·L-1 EDT groups were significantly decreased(P<0.01).Compared with blank control group,the levels of UA,Cr,and BUN in serum of the mice in model group were increased(P<0.01),and the kidney indexes were significantly increased(P<0.01);compared with model group,the levels of UA,Cr,and BUN in serum of the mice in EDT group were decreased(P<0.05 or P<0.01),and the kidney index was significantly decreased(P<0.05 or P<0.01).Compared with blank control group,the number of apoptotic cells in kidney tissue of the mice in model group was increased;compared with model group,the number of the apoptotic cells in kidney tissue of the mice in EDT group was significantly decreased.Compared with blank control group,the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in model group were significantly decreased(P<0.05 or P<0.01),while the expression levels of Bax and MMP-9 proteins were significantly increased(P<0.01);compared with model group,the ratios of p-AKT/AKT and p-PI3K/PI3K and expression level of Bcl-2 protein in kidney tissue of the mice in EDT group were significantly increased(P<0.05 or P<0.01),and the expression levels of Bax and MMP-9 proteins were significantly decreased(P<0.01).Conclusion:The active component of Balanophora involucrata Hook.f.,EDT,has a UA-decreasing effect and may inhibit the apoptosis and alleviate the kidney injury by activating the PI3K/AKT signaling pathway.

Objective To investigate the correlation between neural tube defects (NTDs) and DACT1 gene, and provide the basic data for disease diagnosis and genetic counseling. Methods Blood samples were obtained from 163 NTDs patients and 480 unrelated healthy individuals. Mutation detection of DACT1 gene and DNA direct sequencing was carried out by PCR amplification. Bioinformatics analysis of these mutated loci was performed. Results Six mutations were found in NTDs patients, including 4 missense mutations (p.R45W, p.D142G, p.N356K and p.V702G). But these mutations were not found in 480 healthy individuals. Three mutated amino acid residues (p.45R, p.142D and p.356N) were highly conservative in evolution, and the mutated carriers were female patients, and suffered from anencephaly. Conclusion DACT1 gene mutation may be a risk factor of NTDs in Han population of northern China.

OBJECTIVETo observe abnormal metabolic changes caused by ischemic cerebral apoplexy and the regulating action of Tongsaimai pellets on abnormal metabolism by analyzing the change of small molecules in plasma of ischemic cerebral apoplexy rat. To find the potential biomarkers, and to explore metabolic mechanisms of Tongsaimai pellets.

METHODRat models of middle cerebral artery occlusion was established with electric coagulation, and rats were divided into 4 groups, model group, sham-operation group, Tongsaimai pellets group and positive control group. Tongsaimai pellets and positive control group were orally administrated by 13.2 g x kg(-1) x d(-1) of crude drugs and 32 mg x kg(-1) x d(-1) of Nimodipine respectively, m odel and sham-operation group by equal volume of distilled water for a week. Plasma of model and sham-operation group were collected, and plasma of Tongsaimai pellets and positive control group were collected on the 1st, 3rd , 7th day after administration. Endogenous metabolites of four groups were determined with GC-MS. Partial least squares discriminant analysis (PLS-DA) was applied to analyze multivariate data and set up model, and T-test was used in significant statistical analysis.

RESULTCompared with sham-operation group rats, pyruvic acid, taurine and hydroxyproline obviously increased in model group rats, while lactic acid, glyceric acid, aminomalonic acid, fructose, tryptophan and leucine significantly decreased, so these metabolites were potential metabolic biomarkers. These endogenous metabolites except taurine got restoration in Tongsaimai group rats.

CONCLUSIONAbnormal metabolite level in plasma can be certainly recovered by Tongsaimai pellets, and the treatment of Tongsaimai pellets can be connected with the regulation of related metabolic pathways.

Animals ; Brain Ischemia ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fructose ; blood ; Glyceric Acids ; blood ; Hydroxyproline ; blood ; Lactic Acid ; blood ; Leucine ; blood ; Male ; Malonates ; blood ; Metabolomics ; methods ; Pyruvic Acid ; blood ; Rats ; Rats, Sprague-Dawley ; Stroke ; blood ; drug therapy ; Taurine ; blood ; Tryptophan ; blood

Objective Detrusor overactivity (DO) is one known cause of overactive bladder (OAB) symptoms in benign prostatic hyperplasia (BPH).In this study,OAB symptoms suggestive of DO in BPH were treated with α1-blocker monotherapy or α1-blocker and antimuscarinics add-on therapy,and the efficacy and safety were assessed.Methods BPH patients who suffered from OAB symptoms for at least 3 month were enrolled in a prospective self-control study from August 2010 to April 2012.The inclusion criteria are total international prostate symptom score (IPSS) ≥8,OAB Symptom Score (OABSS) ≥3,OABSS urgency score ≥2,Postvoid residual volume (PVR) ＜ 100 ml,and maximum urinary flow rate (Qmax) ≥ 5 ml/s.All the patients who met the inclusion criteria were treated with α1-blocker ( tamsulosin 0.2 mg/day or doxazosin 4 mg/day) for 2 weeks.After 2 weeks,patients with no symptomatic improvement ( OABSS≥3) underwent pressure-flow test,and those whose Pdet≥ 40 cm H2O and DO presented more than one time were added antimuscarinics (solifenacin 5 mg/day or tolterodine 4 mg/day) for an additional 2 weeks.OABSS,IPSS,QOL,Qmax and PVR were re-evaluated every 2 weeks.Results Ninety-four cases of BPH/OAB patients met the inclusion criteria and completed 4 weeks treatment.The baseline of total OABSS was 7.0 ± 1.3,IPSS was 17.0 ± 1.7,QOL was 5.0 ±0.7,Qmax was (8.8 ±2.5) ml/s and PVR was (86.0 ± 16.5) ml.After 2 weeks treatment with α1-blocker alone,OABSS was 5.2 ± 0.8,IPSS was 14.2 ± 1.9,QOLwas4.O±0.8,Qmaxwas (11.4±2.4) ml/s and PVR was (67.9±12.9) ml.After another2 weeks treatment with α1-blocker plus antimuscarinics,OABSS was 3.1 ± 0.8,IPSS was 11.1 ± 1.9,QOL was 3.1 ± 0.7,Qmax was ( 10.8 ± 2.4) ml/s and PVR was (71.8 ± 11.9 ) ml.Compared with baseline values,OABSS,IPSS,QOL,Qmax and PVR significantly improved (P ＜ 0.01 ) in α1-blocker monotherapy group and α1-blocker plus antimuscarinic group.The improvement of OABSS,IPSS,QOL scores of α1-blocker plus antimuscarinic group were greater than α1-blocker monotherapy group (p ＜ 0.05 ),while Qmax and PVR showed no differences between the two groups.No acute urinary retention (AUR) was deteted.Conclusion Both of α1-blocker monotherapy and α1-blocker with antimuscarinics add-on therapy can improve OAB symptoms.

Objective To investigate the expression of cyclooxgenase 2 (COX-2) and vascular endothelial growth factor (VEGF) in clear cell renal cell carcinoma (CCRCC) and their correlation with Prognosis. Methods EnVision immunohistochemistry was used to determine the expression of COX-2 and VEGF in 80 CCRCC tissues and 20 normal kidney tissues .The relationship between the above marks and prognosis were analyzed. Results The positive rates of COX-2[65.00 % (52/80) vs 10.00 % (2/20), x2= 7.760, P= 0.021]and VEGF[61.25 % (49/20) vs 20.00 (4/80),x2 = 8.870, P= 0.012]were much higher in CCRCC than those in normal kidney. The expression of COX-2 was correlated with TNM stage (x2 = 8.200,P =0.005), histological grade (x2 = 13.860, P = 0.000) and lymph node metastasis (x2 = 6.050, P = 0.001) in CCRCC, but not with age (x2 = 0.560, P = 0.663) and diameter of tumor (x2 = 0.700, P = 0.528). Both COX-2 expression and VEGF expression were associated significantly with prognosis in CCRCC (x2 = 18.280,P = 0.038;x2 = 6.420, P= 0.042, respectively). There was a positive correlation between COX-2 and VEGF in CCRCC (r =0.485, P < 0.01). Conclusion COX-2 is related to prognosis in CCRCC and can be used as prognostic indicators in patients.

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Objective To analyse the operation technique and therapeutic effect of "inserting" uretero-intestinal anastomosis in orthotopic bladder substitution.Methods Thirty-eight patients undergoing orthotopic bladder substitution operations were followed up,and the way of uretero-intestinal anastomosis in all Datients was the "inserting"uretero-intestinal anastomosis.The therapeutic effect was observed by radiation,cystoscopy,pathologic biopsy and blood test.Results The average follow-up time was(3 1.65±14.14)montll8.and the stricture rate was 4%(3/75),but no vesicoureteric reflux was found.The rate of leakage was 0.Nipples were formed at the site of anastomosis under the view of cystoscope,and among the 7 patients whose nipples were taken to be examined by histology,2 cases were intestinal epithelium which were taken at the base of nipple8.while the others were transitional epithelium which were taken at the top of nipples.The renal function of all patients was normal (Cr 54-135 μmol/L,BUN 3.2-9.4 mmol/L).Conclusion "Inserting"uretem-intestinal anastomosis is an ideal antireflux uretero-intestinal anastomosis method.

Objective To study the application of pediculated skin flaps in the treatment of com-plicated long urethratresia. Methods From March 1999 to May 2006, a total of 18 male patients with complicated long urethratresia were treated by using the pediculated skin flaps. The causes of urethratresia were 7 cases of postoperative pelvic fractures with posterior urethral stricture, 4 cases of transurethral intravesical chemotherapy, 3 cases of postoperative bulbar urethral stricture, 2 cases of gonorrhea, and 2 cases of long-time urethral catheter placement. Four cases were urethratresia nf cor-pus penis, 7 cases were anterior urethral obliteration, 7 cases were posterior urethral and anterior ure-thral obliteration. Urethro-perineal fistulas were found in 8 cases, posterior urethrorectal fistulas in 7 cases, false passage formations in 8 cases. The average length of urethratresia was 15.1 cm (range 8. 7 to 23. 0 cm). The urethral scar was rasected, the posterior urethrorectal fistula was repaired, and different kinds of pediculated skin flaps depending on the length of urethratreaia was used. Results All the patients were followed up for 12 to 18 months (mean 14 months). Fifteen patients voided well 3 months postoperatively, none of the urography showed stricture. The mean peak urinary flow rate was 16. 9 ml/s (range from 16. 5 to 21.7 ml/s). Of the other 3 cases, 1 case experienced difficult voi-ding due to the long and circuitous tabularized skin flap but recovered after proper shortening;1 case had restenosis for the infection of anastomosis but voided well after excision and reanastomnsis;1 casehad a urinary fistula resulting from hematoma and infection, but was successfully treated by the neo-plasty of the urinary fistula. The mean peak urinary flow rate was 17.0 ml/s (range 15.0 to 22.0 ml/s) for 17 patients 6 months postoperatively, except for one who experienced genuine urinary incon-tinence. At 9-18 months after operations, the mean peak urinary flow rate was 17.5 ml/s (range 15.8 to 22.5 ml/s) for 17 patients. Conclusion The single-stage urethroplasty based on pediculated skin flaps is a reliable and durable method for complicated long urethratresia.

Objective To evaluate the efficacy of mitomycin (MMC) intravesical chemotherapy for superficial bladder carcinoma by in vitro chemosensitivity using histoculture drug response assay (HDRA).Methods Forty-one cases of superficial bladder transitional cell carcinoma(TCC) were obtained,including 30 males and 11 females with mean age of 55 years.Of them,10 cases were Ta and 31 were T1 according to TNM stage system (UICC 2002) while 9 cases were G1,22 were G2 and 10 were G3 (WHO1973).All cases had no chemotherapy history before operation and were operated to retain bladder.Tumor specimens were cultured by three-dimensional histoculture.HDRA with im-proved MTT assay was utilized for chemosenstivity test of MMC with 1 g/L concentration and 2 hours exposure.Growth inhibition rate (GI) exceeding 70% was defined as high-sensitive while less than 50% GI was defined as insensitive,others were moderate-sensitive.All cases were performed standard intravesical chemotherapy with MMC 40 mg plus 40 mt saline.Every case was followed up every 3 months.The patients were followed up for 5 years or untill recurrence.Results The MMC chem-osensitivity was different among 41 patients.Thirteen cases were insensitive including 1 of Ta,12 of T1 (P=0.009) and 1 of G1,7 of G2,5 of G3(P=0.101).Total recurrence rate was 39%(16/41) af-ter 3 to 5 years follow-up.There were 1 of Ta,15 of T1 (P=0.059) and 10 of G2 6 of G3 (P=0.016).Insensitive group recurrence rate was 77% (10/13) while sensitive group was 21% (6/28,P= 0.004).Patients in sensitive group showed a longer median time(49.2 months) than patients in insen-sitive group (16.5 months,P<0.001) according to Kaplan-Meier analysis with Log-rank test.The MMC chemosensitivity was independent prognostic factor examed by Cox regression analysis (P= 0.008).There was a 78% correlation rate of chemosensitivity by HDRA to clinical effect of MMC in-travesical chemotherapy.Conclusion HDRA could evaluate MMC intravesical chemotherapy for su-perficial bladder TCC,improve therapeutic effect and lower tumor recurrence rate.

BACKGROUND: The studies about cellular cardiomyoplasty (CCM) of bone marrow stromal cell (BMSC) are centered on modeling autologous cell transplantation while the study of myocardium transplantation of allogeneic BMSC is seldom reported domestically.OBJECTIVE: To study the hypothesis that the allogeneic BMSC, after transplanted into the myocardial infarction (MI) regions, can survive, further proliferate, differentiate, and its effects on host hearts.DESIGN: A randomized and controlled trial with experimental animals as subjects.SETTING: Laboratory of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Ten Wistar rats of one-month old, body mass of 100-120 g and unconfined sex were used to culture BMSC while eighty female Wister rats of three-month old, body mass of 200-250 g were used for animal models.METHODS: The experiment was completed in the Laboratory of of Internal Cardiology, Affiliated Union Hospital, Tongji Medical College,Huazhong University of Science and Technology from June to December 2004. ①Eighty rats were used to establish the acute myocardial infarction (AMI) models by ligating the left anterior descending branch of coronary artery, and 45 of them were successful. ② After 4 weeks, the models were divided into 2 groups at random. In the experiment group (n=25), the passaged and uninduced BMSC cultured in vitro were injected into the MI region of the recipients while only medium was injected in the control group (n=20). ③At 4 weeks after implantation,the hemodynamic indexes of recipients' hearts were examined. Then the samples were obtained to detect the survival, differentiation and angiogenesis status of the removal cells.MAIN OUTCOME MEASURES: ①Comparison of hemodynamic indexes in rats of two groups②Results of the implantation cells③Changes of angiogenesis in rats of two groups.RESULTS: Totally 13 rats in the experiment group and 11 in the control group were involved in the result analysis. ①The hemodynamic indexes of rats were significantly improved in the experiment groups compared with the control group [Left ventricle systolic blood pressure (LVSBP): (88.61±5.99),(76.93±4.75) mm Hg, left ventricle end-diastolic pressure(LVEDP): (7.72±1.36),(12.77±2.76) mm Hg, P ＜ 0.05;The maximum changing velocity of LVSBP:(2 365.26±266.31),(2 025.04±230.25) mm Hg/s; The maximum changing velocity of LVDBP:(2 313.26±159.30),(2 140.12±191.03) mm Hg/s]. ②After implanted in the MI region, the allogeneic BMSC passed the acute inflammation period and did not induce the remarkable reject reaction of transplantation. The BMSC in the infarcted region were mainly differentiated into fibroblast. Some cells around the infarcted region were differentiated into endothelial cells, and improved the angiogenesis. ③The number of angiogenesis in and around the transplantation regions was significantly higher in the experiment group than in the control group (P ＜ 0.01).CONCLUSION: The allogeneic BMSC can not form cardiomyocyte in the infarcted region after cell transplantation, and the engendered endothelial cells of blood vessels may promote the angiogenesis after AMI and ameliorate the cardiac function.