Objective This study is about to detect the KCNJ5 gene variations in aldosterone-producing adenoma (APA) with primary hyperaldosteronism (PA),and to investigate the association of the KCNJ5 gene missense mutations with APA and PA.Methods A total of 46 APA tumors and their clinical characteristics were collected from Hypertension Center of the People's Hospital of Xinjiang Uygur Autonomous Region,and all the tumors were confirmed by pathology.All the samples of the coding region segments of KCNJ5 were detected by PCR and direct DNA sequencing to compare the different missense mutations in the tumor cells and peripheral blood cells and to analyze the association between the genotype and phenotype.Results Three missense mutations were found in 46 patients with APA:c.451G ＞ C/A (p.G151R) (5/46),c.503T ＞ G (p.L168R) (4/46),c.830T ＞ A (p.S209T) (12/46).S209T,as an unreported somatic mutation was observed.There were no missense mutations detected in the peripheral blood.Sex,age,systolic blood pressure,diastolic blood pressure,duration of hypertension,plasma potassium,urine potassium,aldosterone,plasma renin activity,aldosterone,and plasma renin activity ratio,as well as the rate of positive family history were compared between the mutants and wild-types.Systolic blood pressure and plasma potassium level among G151 R,L168R,and S209T had statistical differences.Systolic blood pressure in G151R was significantly higher than the other two groups,while the plasma potassium level was significantly lower than the other groups.Conclusions Three missense mutations were found in 46 patients with APA.The subjects with mutations had more serious condition than those without mutations after comparing their clinical phenotype.Besides,other different subtypes may exist in PA.

OBJECTIVETo observe the effect of Bushen Huoxue Granule (BHG) on dopamine (DA) neurotransmitter and dopamine transporter (DAT) in the brain of patients with Parkinson's disease (PD) as an adjunctive therapy.

METHODSNinety-four PD patients were randomly assigned to two groups, 47 in each group. Madopar was given to all as the basic treatment group. The placebo was given to those in the control group while BHG was given to those in the treatment. The therapeutic course for all was three months. Before and after treatment DA levels in the brain of patients were detected by encephalofluctuograph (EFG) technique. Changes of DAT in the striatum of patients in the treatment group were detected by positron emission tomography (PET) and region of interest (ROI) analysis.

RESULTS(1) Before treatment the DA level was lower in the two groups than the normal value, showing significant difference (P < 0.01), but with no significant difference between the two groups (P > 0.05). After treatment the DA level obviously increased in the two groups, showing significant difference from that before treatment (P < 0.01). No significant difference existed in the DA level in the two groups when compared with the normal value (P > 0.05), but with significant difference between the two groups (P < 0.05). Better results were obtained in the treatment group than in the control group. (2) The DAT radioactive accumulation inside the striatum increased obviously in the treatment group after treatment. ROI analysis showed the total ratio of striatum/cerebellum before and after treatment was 1.86 +/- 0.32 and 2.61 +/- 0.53 respectively, showing statistical difference (P < 0.05).

CONCLUSIONBHG could improve the DA level of PD patients, and increasing DAT contents in the striatum, thus playing a role in effectively treating PD.

Aged ; Brain ; metabolism ; Dopamine ; metabolism ; Dopamine Plasma Membrane Transport Proteins ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; drug therapy ; metabolism ; Phytotherapy

Objective To establish an HPLC method for the content determination of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from different habitats.Methods The chromatographic separation was performed on an Agilent C18 column (4.6 mm×150 mm, 5μm) with mobile phase of acetonitrile-water solution with gradient elution at the flow rate of 0.8 mL/min; the detection wavelength was 208 nm; the column temperature was 30℃; the injection volume was 20μL.Results Protodioscin showed a good linear relationship among the range of 1.73–8.64 μg (r=0.999 6), with the average recovery of 101.98% (RSD=1.53%); Diosgenin showed a good linear relationship among the range of 1.03–8.20μg (r=0.999 1), with the average recovery of 101.60% (RSD=2.41%). The contents of protodioscin and diosgenin in Dioscoreae Hypoglaucae Rhizoma from 10 different habitats were in the range of 0.89%–2.24% and 0.75%–3.22%, respectively.Conclusion The method is simple, accurate and with repeatability, which can be used as quality control method of Dioscoreae Hypoglaucae Rhizoma.

Objective To study the inhibitory effect of antigliomatin on the expression of vascular endothelial growth factor(VEGF) in rat C6 brain glioma cells.Methods The rat brain glioma cells was cultivated with the stationary culture technique.The different rat C6 brain glioma cells groups were treated with 2,4,8,and 16 ng?L-1 antigliomatin for 72 h in vitro and then the inhibitory effect of antiglioma on the expression of VEGF was observed with immunohistochemical method.Results The cytoplasm and(or) nucleus of most VEGF positive cells were stained to yellow brown.VEGF expressed significantly in cytoplasm of the rat C6 glioma cells in control group.The density of the VEGF positive cells was decreased in glioma compared with control group after treated with antigliomatin for 72 h(P

Objective To evaluate the value of 99Tcm-MDP uptake by left femoral neck for diagnosing osteopomsis.Methods A total of 58 cases (23 males,35 females,mean age:(66.15±8.45) years) with spondyloarthmpathies from May to December of 2012 were selected.Serum concentrations of type Ⅰ collagen telopeptide (sCTX-1) and bone ALP (BALP) were determined.All patients underwent dual-energy X-ray absorptiometry (DXA) to detect bone mineral density (BMD).According to the T scores,patients were divided into 2 groups:normal group (NG) (T＞-1.0) and osteoporosis group (OG) (T≤-2.5).99TcmMDP bone scan was further performed.The average radioactive ratio of the left femoral neck to the medial soft tissue of left femur (T/N) was measured.Data differences between the 2 groups were compared by twosample t test and Pearson correlation analysis.Results According to BMD,13 patients (7 males,6 females) were included in NG and 28 patients (10 males,18 females) were included in OG.The mean ages of OG and NG were significantly different ((68.82± 10.41) years vs (62.46± 11.77) years; t =3.560,P＜0.05).The BMD of left femoral neck in OG was significantly lower than that in NG ((0.67±0.08) g/cm2 vs (0.91±0.10) g/cm2 ; t=9.917,P＜0.01).Although BALP level of OG was significantly higher than that of NG ((35.92±11.58) U/L vs (22.38±6.34) U/L; t=-3.397,P＜0.05),no significant difference was observed on sCTX-1 between the 2 groups (t=-0.463,P＞0.05).T/N ratio of OG (11.63±6.22) was higher than that of NG (9.74±4.44) (t =-3.027,P＜ 0.05).There were significant correlations between the T/N ratio of the left femoral neck and the sCTX-1 and BALP concentrations (r=0.376,0.483,both P＜0.01).No correlations between the T/N ratio of the left femoral neck and age,BMI and BMD were observed (r=-2.031,-0.017,0.134,all P＞0.05).Conclusion The uptake ratio of the left femoral neck in 99Tcm-MDP bone scan could evaluate the metabolism of bone,and it is useful for the early diagnosis of osteoporosis.

Objective To study the genotypes of Mycobacterium tuberculosis(M.tuberculosis)strains isolated from Jiangsu province and to explore the relationship between the 'Beijing family' and the drug resistance of M.tuberculosis.Methods Two hundred and sixty M.tuberculosis strains were isolated from 30 drug surveillance sites in Jiangsu province.Susceptibility of the isolates to the first-line antituberculosis drugs(isoniazid,streptomycin,rifampicin and ethambutol)was tested by using the proportion method.Molecular typing of M.tuberculosis strains was determined by Spoligotyping and analyzed with BioNumerics software.Results Based on Spoligotyping fingerprint,260 strains showed 34 different genotypes,including 27 exclusive genotypes and 7 shared genotypes.These strains could be clustered into two groups:the Beijing family(80.4％,209/260)and the Non-Beijing family(19.6％,51/260).Data from logistic regression analysis revealed that infection by the Beijing family was related to an increased risk of multi-drug resistant M.tuberculosis,with the OR(95％ CI)of 11.07(1.45-84.50).Non-Beijing families including T1,T2,H3,H4,CAS,LAM,U and MANU2 families were also found.Among them,the CAS,LAM and MANU2 families were first reported in Jiangsu province.Conclusion It was revealed that the marked gene polymorphisms did exist in M.tuberculosis strains.The Beijing family had been the predominant strain circulating in Jiangsu province,which might be related to multi-drug resistant M.tuberculosis strains.

A series of cycloberberine derivatives were designed, synthesized and evaluated for their anti-cancer activities in vitro. Among these analogs, compounds 6c, 6e and 6g showed strong inhibition on human HepG2 cells. They afforded a potent effect against DOX-resistant MCF-7 breast cells as well. The primary mechanism showed that cell cycle was blocked at G2/M phase of HepG2 cells treated with 6g using flow cytometry assay. It significantly inhibited the activity of DNA Top I at the concentration of 0.1 mg mL-1. Our results provided a basis for the development of this kind of compounds as novel anti-cancer agents.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Berberine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type I ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Hep G2 Cells ; Humans ; MCF-7 Cells ; Molecular Structure ; Structure-Activity Relationship

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.

OBJECTIVETo observe the clinical curative effect of Chinese medicine Bushen Huoxue Granule(补肾活血颗粒, BHG) on Parkinson's disease (PD) patients with depressive state.

METHODSSixty-two PD patients with depressive state were randomly assigned to two groups by using a random number table, 31 in each group. Madopar was given to all as the conventional treatment. The fluoxetine hydrochloride dispersible tablet was given to the patients in the control group and BHG was given to those in the treatment group. The therapeutic course for all was 12 weeks. Before and after treatment, Hamilton depression rating scale (HAMD) was applied to judge the curative effect, and the changes of cerebral neurotransmitters levels in the brain of patients were detected by encephalofluctuograph technique.

RESULTSThe scores of HAMD in the two groups were decreased markedly after 12-week treatment. It was lower in the treatment group than that in the control group with significant difference (P<0.01). The contents of norepinephrine (NE) and 5-serotonin (5-HT) in the PD patients were obviously lower than normal value. There was no significant difference between the two groups before treatment (P>0.05). The contents of NE and 5-HT were all increased in the two groups after treatment (P<0.05), with significant differences between the two groups (P<0.01).

CONCLUSIONBHG could increase the contents of NE and 5-HT in PD patients' brain to improve the depressive state of PD patients.

Aged ; Brain ; drug effects ; metabolism ; Depression ; complications ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Norepinephrine ; metabolism ; Parkinson Disease ; complications ; drug therapy ; Serotonin ; metabolism ; Treatment Outcome