ObjectiveTo study the influence of anti-tuberculosis drugs on mitochondrial function in mice hepatocytes and to explore the mechanism of the anti-tuberculosis drugs induced liver injury.Methods A total of 150 mice were randomized into five groups:control group (C group),rifampin (RFP) group,isoniazid (INH) group,pyrazinamide (PZA) group and three antituberculosis drug combination group (MIX).The mice were administered intragastrically with 0.9 ％ NaC1 solution or RFP 135 mg · kg-1 · d-1 or INH 90 mg · kg-1 · d-1 or PZA 315 mg · kg-1 · d-1 or RFP+INH+ PZA (135±90+315) mg · kg-1 · d-1 once a day.Ten mice in each group were sacrificed at day 3,7 and 15 of administration,respectively.The following parameters in each group were monitored.the concentration of malondialdehyde (MDA),the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in mitochondrion of hepatocytes and the concentration of 8-hydroxydeoxyguanosine (8-OH-dG) in mitochondrial DNA (mtDNA).The data were analyzed by one-way ANOVA or rank sum test.Results Along with the prolonged medication duration,the concentrations of MDA all gradually increased in RFP group (Z=6.020,P=0.049),IN H group (Z=10.220,P=0.006) and MIX group (Z=7.460,P=0.024),whereas the activity of SOD significantly decreased in RFP group (F=6.751,P =0.011 ) and MIX groups (F=4.891,P =0.041 ) compared with control group and PZA group.Meanwhile,the activity of GSH-PX was significantly lower in RFP group compared to the other groups (F=32.445,P＜0.01).The changes of other parameters didn't show meaningful trend.The concentrations of 8-OH-dG in mtDNA also increased in all treated groups,and those were all significantly increased in RPF group (F=6.602,P＜0.01 ),PZA group (F=5.927,P＜0.01) and MIX groups (F=7.974,P＜0.01).Conclusions Antituberculosis drugs can induce higher MDA concentration in mitochondrion and higher 8-OH-dG concentration in mtDNA,while result in lower activities of SOD and GSH-PX.The liver damage tends to become more severe along with the prolonged medication duration.The combination of three antituberculosis drugs could aggravate the damage of mitochondrion in mice hepatocytes.

OBJECTIVEThe purpose of this study was to investigate the relation between the Arg389Gly polymorphism of the beta(1)-AR gene and chronic heart failure (CHF) and to evaluate the effect of this polymorphism on clinical response to beta-adrenoceptor blockade (bisoprolol) in patients with CHF.

METHODSOne hundred and ten patients with stable CHF receiving basic therapy for heart failure were included. Before initiation and 3 months after the maximal tolerated dose of bisoprolol was reached, all indices (including BP, HR, LAD, LVEDD, LVESD, LVEF, BNP level, 6 min walk distance) were measured and compared with the Arg389Gly genotypes, which identified by PCR-restriction fragment length polymorphism analysis. We also determined the Arg389Gly genotypes in 100 healthy control subjects, and compared the distribution of Arg389Gly genotypes with that in CHF.

RESULTSNo difference was observed between the two groups in any of the three genotypes (CC, CG and GG). The prevalences of the three genotypes in normal subjects and patients with CHF were Arg389Arg 0.53 vs. 0.51, Arg389Gly 0.40 vs. 0.40, Gly38Gly 0.07 vs. 0.09, respectively. After 3 months of bisoprolol usage, a significant improvement in LVEF was observed in CC group, which increased from (36.7 +/- 8.63)% to (44.1 +/- 9.53)%, CG group, from (35.76 +/- 8.39)% to (42.90 +/- 9.41)%, but not GG group, from (36.00 +/- 5.66)% to (37.33 +/- 5.64)%. The improvement in BNP was also observed in CC [from (502.93 +/- 160.80) ng/L to (325.26 +/- 135.63) ng/L], CG [from (525.76 +/- 157.66) ng/L to (331.79 +/- 133.97) ng/L], but not GG [from (505.33 +/- 125.07) ng/L to (429.67 +/- 182.39) ng/L]. Arg389-homozygous patients showed a substantially greater improvement in LVEF and BNP, compared with Gly389-homozygous patients (all P < 0.01).

CONCLUSIONSThere was no difference in the prevalence of the three genotypes between healthy and CHF subjects. The Gly389 polymorphism of the beta(1)-AR gene was not associated with an increased risk of CHF. The Arg389 variant of the beta(1)-AR gene was associated with a greater response to bisoprolol than that of the Gly389 variant in patients with CHF.

Adrenergic beta-Antagonists ; therapeutic use ; Adult ; Aged ; Aged, 80 and over ; Bisoprolol ; therapeutic use ; Case-Control Studies ; Female ; Follow-Up Studies ; Heart Failure ; drug therapy ; genetics ; physiopathology ; Humans ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood ; Polymorphism, Genetic ; Receptors, Adrenergic, beta-1 ; genetics

OBJECTIVETo explore clinical effects of manipulative reduction with minimally invasive percutaneous plate osteosynthesis in treating distal tibiofibular fractures.

METHODSFrom 2009 to 2011, 60 patients with distal tibiofibular fractures were treated by manipulative reduction with minimally invasive percutaneous plate osteosynthesis. Among them, there were 32 males and 28 females aged from 14 to 70 years old with an average of 41.22 +/- 2.06. According to AO classification of fractures,5 cases were type A1, 22 cases were type A2,21 cases were type A3 and 12 cases were type C1. Operation time, blood loss,time of callus and fracture healing were observed, Mazur scoring of ankle joint were used to evaluate therapeutic.

RESULTSFifty-eight incisions were healed at stage I ,and 2 cases were infected at distal tibial. Operation time was with an average of (62.34 +/- 5.66) min ranged 45 to 90 min;blood loss was 30 to 150 ml with an average of (80.57 +/- 5.59) ml;formation of callus appeared from 4 to 12 weeks,with an average of (8.24 +/- 2.06) weeks, and fracture healing time was from 3 to 6 months, with an average of (4.50 +/- 1.13) months. According to Mazur scoring of ankle joint 40 cases got excellent results, 18 good, and 2 fair.

CONCLUSIONManipulative reduction with minimally invasive percutaneous plate osteosynthesis can obtain reliable fixation. It is a good choice of treating distal tibiofibular fractures by protecting blood supply of fractures.

Adolescent ; Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVE: To investigate the polymorphism of the TPH gene T3792A locus in Han ethnic group of north China and its application value in forensic science. METHODS: The polymorphism of T3792A locus of the TPH gene was analyzed by the ASPCR of blood samples from 173 unrelated individuals of north Chinese Han population. RESULTS: The distribution of the T3792A locus polymorphism of the TPH gene in Han ethnic group of north China followed the Hardy-Weinberg law, with the allele A and T gene frequency of 0.486 and 0.514, respectively. CONCLUSION The TPH gene T3792A locus shows a very good genetic polymorphism, and may be applied to individual identification and paternity testing.
Asian People/genetics* ; China/ethnology* ; Forensic Genetics ; Gene Frequency ; Humans ; Paternity ; Polymorphism, Genetic ; Tryptophan Hydroxylase/genetics*

BACKGROUNDBcl-2, the anti-apoptotic protein is overexpressed in the majority of gastric cancers and associated with its pathogenesis. To better understanding of the role of Bcl-2, RNA interference (RNAi) was used to inhibit Bcl-2 expression in the human gastric cancer cells in vitro and in vivo.

METHODSBcl-2 small interfering RNA (siRNA) was transfected into human gastric cancer cells SGC-7901, and Bcl-2 expression was monitored by real-time polymerase chain reaction (PCR) and Western blot. Cell proliferation, apoptosis, and telomerase activity were examined by MTT, flow cytometry, and TRAP assay, respectively. Gastric cancer cells treated with 100 nmol/L Bcl-2 siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed.

RESULTSBcl-2 siRNA significantly inhibited the expression of Bcl-2 in human gastric cancer cells at both mRNA and protein levels in a time- and dose-dependent manner. Bcl-2 siRNA also decreased telomerase activity (by 78.76%) and increased the rate of apoptosis (by 37.47%). SGC-7901 cell growth was also significantly suppressed in vivo and in vitro.

CONCLUSIONSBcl-2 expression knockdown suppressed the growth of gastric cancer cells. Thus, Bcl-2 may play a very important role in carcinogenesis of gastric cancer and its knockdown may offer a new potential gene therapy approach for human gastric cancer in future.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; pathology ; therapy ; Transfection

OBJECTIVETo evaluate the safety and efficacy of individual cylindrical abdominoperineal resection (CAPR) for locally advanced low rectal cancer.

METHODSFrom June 2011 to February 2012, 11 patients with locally advanced low rectal cancer underwent individual CAPR. There were 7 male and 4 female patients, aged from 32 to 74 years with a median of 64 years. Forty-seven patients underwent classic CAPR from January 2008 to February 2012. Preoperative and postoperative parameters such as clinical information of patients, tissue morphometry and complications were compared.

RESULTSIn the individual surgical group, 6 patients were treated with one side levator ani muscle totally or partially reserved, 3 patients with sacrococcyx reserved, and 2 patients with dissection close to the anterior rectal wall. Compared with classical surgery, the individual surgical specimens of horizontal section area ((2197 ± 501) mm(2)) and intrinsic muscle layer outer area ((1722 ± 414) mm(2)) were small, but the difference was not statistically significant (P = 0.150 and 0.167). The operative time, intraoperative blood loss, circumferential resection margin, total cross sectional tissue area, cross sectional tissue area outside the muscularis propria and bowel perforation rate between the two groups were not significantly different. Individual CAPR showed less incidence of chronic perineal pain (2/11, χ(2) = 6.116, P = 0.013) and sexual dysfunction (2/9, χ(2) = 4.412, P = 0.036) compared with classic CAPR.

CONCLUSIONSIndividual CAPR has the potential to reduce the risk of chronic perineal pain and sexual dysfunction without influencing the radical effect when compare with classic CAPR for the treatment of low rectal cancer.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Perineum ; surgery ; Postoperative Complications ; epidemiology ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; surgery ; Survival Rate ; Treatment Outcome

OBJECTIVETo develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.

METHODSThe gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Vero cell. CPE was observed and viral gene viral antigen were detected.

RESULTSThe gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA.

CONCLUSIONThe method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.

Animals ; Cercopithecus aethiops ; DNA-Directed RNA Polymerases ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; physiology ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; HeLa Cells ; Humans ; Plasmids ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Proteins ; genetics ; metabolism ; Virus Replication

BACKGROUNDAlthough various systemic and local factors such as abnormal carbohydrate or calcium metabolism, aging, and hormonal disturbances have been suggested as causes of ossification of the posterior longitudinal ligament (OPLL), the etiology of OPLL is not fully understood. The purpose of this study was to investigate whether bone morphogenetic protein (BMP)-2 is a candidate gene to modify the susceptibility of OPLL and the mechanism of signal transduction in ossification.

METHODSA total of 420 OPLL patients and 506 age- and sex-matched controls were studied. The complete coding sequence of the human BMP-2 gene was analyzed using polymerase chain reaction (PCR) and direct sequencing. All single nucleotide polymorphisms (SNPs) were detected and genotyped. BMP-2 expression vectors containing positive polymorphisms were constructed and transfected into the C3H10T1/2 cells. The expression of BMP-2 and the Smad signal pathway in positive cell clones were detected by Western blotting. The alkaline phosphatase (ALP) activity was determined using quantitative detection kits.

RESULTSThe frequencies for the 109T > G and 570A > T polymorphisms were different between the case and control groups. The "TG" genotype in 109T > G polymorphism is associated with the occurrence of OPLL, the frequency of the "G" allele is significantly higher in patients with OPLL than in control subjects (P < 0.001). The "AT" genotype in 570A > T polymorphism is associated with the occurrence of OPLL, the frequency of the "T" allele is significantly higher in patients with OPLL than in control subjects (P = 0.005). Western blotting analysis revealed that the expression of P-Smad1/5/8 protein transfected by wild-type or mutant expression vectors were significantly higher than control groups (P < 0.05), but there was no statistical difference in each experimental group (P > 0.05). The expression of Smad4 protein transfected by wild-type or mutant expression vectors was significantly higher than control groups (P < 0.05). The expression of Smad4 protein transfected by pcDNA3.1-BMP2 (109G) and pcDNA3.1-BMP2 (109G, 570T) was significantly higher than the other experimental groups (P < 0.05). The increase in ALP activity has been detected in pcDNA3.1-BMP2 (109G) and pcDNA3.1-BMP2 (109G, 570T) transfected cells up to 4 weeks after stable transfection. Activity of ALP was (30.56 ± 0.46) nmol×min(-1)×mg(-1) protein and (29.62 ± 0.68) nmol×min(-1)×mg(-1) protein, respectively. This was statistically different compared with the other experimental groups (P < 0.05).

CONCLUSIONSBMP-2 is the predisposing gene of OPLL. The "TG" genotype in the 109T > G and the "AT" genotype in the 570A > T polymorphisms are associated with the occurrence of OPLL. The 109T > G polymorphism in exon-2 of the BMP-2 gene is positively associated with the level of Smad4 protein expression and the activity of ALP. The Smad mediated signaling pathway plays an important role during the pathological process of OPLL induced by SNPs of BMP-2 gene.

Adult ; Aged ; Bone Morphogenetic Protein 2 ; genetics ; Cells, Cultured ; Female ; Humans ; In Situ Hybridization ; Longitudinal Ligaments ; metabolism ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Signal Transduction ; genetics ; physiology ; Smad Proteins ; metabolism ; Spine ; metabolism

OBJECTIVETo investigate the effect of PI-3K and p38MAPK signal pathways on the cyclooxygenase-2 (COX-2) expression induced by the epidermal growth factor (EGF) in PC-3 cells.

METHODSPC-3 cell proliferation was detected by methylthiazolyl tetrazolium (MTT) assay after stimulated by EGF (0 microg/L), EGF (10 microg/L), EGF (10 microg/L) + LY294002 (20 micromol/L) and EGF (10 microg/L) + SC203580 (20 micromol/L), and so was the COX-2 expression in the PC-3 cells by RT-PCR and Western blot assay after stimulated the same way for 24 hours. ELISA was used to determine the changes of PGE2 in the culture medium.

RESULTSLY294002 and SC203580 signficantly inhibited PC-3 cell proliferation (P < 0.05), COX-2 expression and PGE2 production after EGF stimulation (P < 0.05).

CONCLUSIONEGF can stimulate PC-3 cells into proliferation and induce COX-2 mRNA and the upregulation of its protein expression, while LY294002 and SC203580 can inhibit EGF from the above effects on PC-3 cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; genetics ; metabolism ; Dinoprostone ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; pharmacology ; Gene Expression ; drug effects ; Humans ; Male ; Phosphatidylinositol 3-Kinases ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo evaluate the effectiveness and safety of the combination chemotherapy of capecitabine (X) with fractionated administration of cisplatin (C) in Chinese patients with advanced gastric cancer (AGC).

METHODS141 patients with AGC were enrolled between July 2002 and August 2004. All patients had measurable tumor according to the criteria of RECIST, Karnofsky performance status > or = 60, adequate bone marrow, renal and hepatic functions. Prior radiotherapy or adjuvant chemotherapy was not permitted. Patients received oral administration of capecitabine at a dose of 1000 mg/m(2) twice a day on D1-D14, and intravenous infusion of fractionated cisplatin at a dose of 20 mg/m(2)/day on D1-D5. The regimen was repeated every 3 weeks, totally for 6 cycles.

RESULTSOf the 141 evaluable patients, there were 104 men and 37 women, with a median age of 54 years (range, 23 - 80 years). Metastases before chemotherapy were detected in lymph nodes (46.8%), liver (40.4%), lung (5.7%) and other area (10.6%). The median treatment duration was 6 cycles (range, 3 - 6 cycles). The objective response rate (RR) was 36.2% (51/141). The median follow-up period was 17.5 months. The median time to progress (TTP) was 9.0 months, and the median overall survival (OS) was 12.0 months. The most common treatment-related adverse events (grade 3/4) were: hand-foot syndrome (HFS) (2.1%), leucopenia (0.7%), abnormal alanine transaminase elevation (2.8%). There was no treatment-related death.

CONCLUSIONCapecitabine combined with fractionated cisplatin is highly effective and well tolerated as a first-line treatment for advanced gastric cancer, with comparable results to 5-Fu plus cisplatin combination therapy.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Capecitabine ; Cisplatin ; administration & dosage ; adverse effects ; Deoxycytidine ; administration & dosage ; adverse effects ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; adverse effects ; analogs & derivatives ; Follow-Up Studies ; Foot Dermatoses ; chemically induced ; Hand Dermatoses ; chemically induced ; Humans ; Leukopenia ; chemically induced ; Liver Neoplasms ; drug therapy ; secondary ; Lung Neoplasms ; drug therapy ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Remission Induction ; Stomach Neoplasms ; drug therapy ; pathology ; Survival Rate ; Vomiting ; chemically induced ; Young Adult