Objective To evaluate the effects of anisodamine on apoptosis in cardiomyocytes and inflammatory response in overtrained rats. Methods Twenty-four male Wistar rats, weighing 200-220 g, were randomly divided into 3 groups ( n = 8 each) : control group (group C) , overtraining group (group O) , anisodamine group (group A) . The model of overtraining-induced acute heart injury was established by exhausting swimming. Anisodamine 10 mg/kg was given intraperitoneally 20 min before overtraining in group A. Blood samples were taken at 6 h after overtraining for measurement of serum CK-MB activity. The rats were then sacrificed and myocardial tissues taken for determination of TNF-α content and NF-κB activity (by immunohistochemistry) . The apoptosis rate was detected by flow cytometry. Results The CK-MB activity, apoptosis rate, TNF-α content and NF-κB activity were significantly higher at 6 h after overtraining in groups O and A than in group C, while lower at 6 h after overtraining in group A than in group O ( P < 0.05) . Conclusion Anisodamine can inhibit apoptosis in cardiomyocytes by reducing inflammatory response in overtrained rats.

Leukemia inhibitory factor (LIF) plays important roles in varieties of biological processes. This factor is highly conserved in mammalian animals and only one heterozygous LIF mutation was reported to cause the infertility of women. A LIF mutation was generated and the evidences were provided that the mutation of mature LIF at the 29th amino acid totally abolished its functions, including stimulation of STAT activation assayed by Luciferase reporter gene expression and EMSA experiments. In addition, the mutated LIF failed to inhibit the proliferation of M1 cells. The data indicated that the mutation of LIF did not have a dominant negative effect but lost the biological functions, suggesting that the 29th amino acid is critical for maintaining the activities of LIF.

Objective To probe the effects of nuclear factor kappa B (NF-?B) on cell apoptosis and left ventricular segmental function in acute ischemia repeffusion in dogs.Method Twenty-four dogs were randomly divided into three groups:without left anterior artery (lAD) ligation group (C group),LAD was occluded 30 min following reperfusion 120 minutes in isehemical reperfusion group (IR group),and dogs were administered with PDTC before LAD ligation in ischemical reperfusion plus pyorrole dithitocarbamate group (PDTC group).The left ventricular segmental function was detected by echo cardiography using strain rate anlysis software.EF measured by Simpson's method.Cardiac myocyte apoptosis numbers were determined by terminal deoxynudeotidy transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling (TUNEL).lmmunohistochemistry and western-blot anylysis of NF-?B protein expression.Results NF-?B was obviously expression on injury myocardium of IR group,and increased significantly in contrast to control group (P0.05)Conclusions NF-?B might play an important role in acute myocardial ischemia reperfusion.PDTC reduces myocardial iscbemia/repeffasion injury by preventing expression of factor NF-?B.

Objective To explore the initial clinical and imaging characteristics of basal ganglia germinoma in children.Methods Four patients with basal ganglia germinoma were reported.Their clinical features,laboratory findings,radiological manifestations,treatment and outcome were analyzed.They recieved radiation therapy and chemotherapy after diagnosis.All patients were clinically diagnosed,according to the results of low-dose cranial irradiation.The outcomes were followed up for 2 years.Results All patients were male and school-aged(9-13 years) children.The course of the disease ranged from 5 to 13 months.All patients were presented with slowly progressive hemiparesis,and 2 cases of them were presented with cognitive decline and psychosis.Seizure occurred in 2 patients.The serum ?-human chorionic gonadotropin(?-hcG) level was significantly increased in 2 patients(30.16 IU/L and 77.85 IU/L,respectively),and mildly elevated in 1 patient(4.29 IU/L),while serum ?-hcG level in another case was within normal control range.MRI demonstrated mildly high intensity in the left or right basal ganglia on T1-weighted and T2-weighted images without remarkable occupying lesion.Ipsilateral hemiatrophy of the hemisphere and midbrain was also noted.Inhomogeneous Gd-DTPA enhancement was observed.All patients had been treated with radiation therapy and chemotherapy.During 2 years follow up,significant improvement was observed in all patients after therapy,imaging lesions disappeared and the elevated ?-hcG level of those elevated before therapy returned to normal.Conclusions Early diagnosis and treatment for basal ganglia germinoma are critically important to improve the prognosis.In young male patients with progressive hamiparesis,basal ganglia germinoma should be considered for differentiation,if abnormal high intensity signals in basal ganglia on T1-weighted and T2-weighted image with ipsilateral hemiatrophy of the hemisphere are demonstrated on MRI,even without occupying effect.

Abstract: Objective To explore the influencing factors of serum HBeAg loss in patients with chronic hepatitis B (CHB) and and provide evidence for effective treatment of CHB. Methods A follow-up cohort of HBeAg-positive CHB patients was established in the the Infectious Diseases Outpatient Clinic of hospital. Regular follow-up and laboratory test indicators were collected to analyze the changes of serum HBeAg in HBeAg-positive CHB patients during the follow-up period. The subjects were divided into the case group (serum HBeAg loss) and the control group (serum HBeAg not loss) according to whether serum HBeAg loss occurred. The baseline data characteristics of the two groups were analyzed and compared, and the influencing factors of serum HBeAg loss were analyzed by Cox univariate and multivariate regression. Results A total of 634 HBeAg-positive CHB patients were enrolled, with a total follow-up of 2 570.01 person-years. Among them, 237 cases of serum HBeAg loss occurred, with the mean follow-up time of 40.92 months, and the rate of HBeAg loss was 9.22/100 person-years. There were significant differences in HBV family history, antiviral therapy, baseline WBC, PLT, ALT, AST, T˗Bil, GGT, AFP, quantitative HBsAg and quantitative HBeAg between serum HBeAg loss group and serum HBeAg not loss group (P<0.05). Cox regression analysis showed that family history of HBV (HR 0.68, 95％CI:0.50-0.92, P=0.012), ALT (HR2.06, 95％CI:1.52-2.79, P<0.001), quantitative HBsAg (HR 0.68, 95％CI:0.48-0.95, P=0.024), quantitative HBeAg (HR 0.48, 95％CI:0.31-0.74, P=0.001) were independent influencing factors for HBeAg loss in HBeAg-positive CHB patients. Conclusions HBeAg-positive CHB patients without family history of HBV, initial ALT≥80 U/L, quantitative HBsAg<1 000 IU/ml, quantitative HBeAg<1 000 C.O.I are more likely to have serum HBeAg loss.

Objective: To study the protective effect of Ascorbic acid (AA) on the injury of nickel-exposed mouse embryonic fibroblasts (NIH/3T3) . Methods: A model of damage induced by 50 μg/mL nickel refining dust was established to determine the relative survival rate of cells, superoxide dismutase (SOD) , lactate dehydrogenase (LDH) and glutathione peroxidase. (GSH-Px) activity, hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) content, and p53 (wild-type) , Bcl-2 protein expression. To investigate the protective effect of different doses of ascorbic acid (25, 50, 100 mmol/L) on nickel-refined dust-induced NIH/3T3 cell injury. Results: The study showed that ascorbic acid Ⅲ group can make the NIH/3T3 cell survival rate increased significantly; Apoptosis rate was reduced; The vitality of SOD and GSH-Px increased significantly, and the difference was statistically significant (P<0.05) . At the same time, the level of MDA and H₂O₂ and the activity of extracellular LDH enzyme were significantly reduced, and the difference was statistically significant (P<0.05) . The results showed that nickel refining dust induced cell damage through up-regulation of p53 protein and down-regulation of Bcl-2 protein expression; ascorbic acid interventions, the expression level of Bcl-2 protein in ascorbic acid II and III groups was higher than that of nickel refining dust group, and the difference was statistically significant (P<0.05); The expression level of p53 protein in each dose group of ascorbic acid was lower than that of nickel refined dust group, and the difference was statistically significant (P<0.05). Conclusion With the increase of concentration of ascorbic acid, oxidative damage levels, antioxidant enzyme levels, reduce cell apoptosis, reduce expression of p53, increased expression of Bcl-2. It showed that ascorbic acid had protective effect on NIH/3T3 cell injury induced by nickel refining dust.

Objective To establish the finite element model of uterosacral ligament (USL) and cardinal ligament (CL) and analyze the stress distribution and deformation with USL and CL under different working conditions. Methods Patients with stage Ⅲ-Ⅳpelvic organ prolapse (POP) and healthy female volunteers were selected for research subject, and divided into anterior uterus group and posterior uterus group. Two POP patients and two volunteers were selectd into the anterior uterus group and posterior uterine group respectively. Pelvic MRI scan was performed in two groups. Based on the original MRI data sets, the finite element model of USL and CL was constructed by using the software such as the Mimics, and the stress distribution and deformation of USL and CL were simulated. Results Under the premise of the elastic modulus fixed and three different working conditions such as 60 cmH2O, 99 cmH2O and 168 cmH2O (1 cmH2O=0.098 kPa) with abdominal pressure generated by maximum Valsalva maneuver, according to the present conditions and the simulation, the trend was analyzed: the stress and deformation of the uterus, anterior vaginal wall, USL and CL in two groups were mainly distributed in the middle and lower part of the anterior vaginal wall or the ligament and the cervix-vagina junction, the maximum stress and the maximum displacement were mainly concentrated in the lower region of the anterior vaginal wall. With increasing of abdominal pressure generated by the maximum Valsalva maneuver, the maximum stress values of the POP patient in anterior uterus group under three different working conditions were: 0.027 9, 0.046 0, 0.078 0 MPa, and the maximum displacement values were: 9.145 5, 15.090 0, 25.607 0 mm. The maximum stress values of the volunteer in anterior uterus group under three different working conditions were:0.012 6, 0.020 8, 0.035 3 MPa, and the maximum displacement values were: 1.816 7, 2.997 5, 5.086 7 mm. The maximum stress values of the POP patient in posterior uterine group under three different conditions were: 0.069 4, 0.114 6, 0.194 5 MPa, and the maximum displacement values were:11.658 0, 19.236 0, 32.643 0 mm. The maximum stress values of the volunteer in posterior uterus group under three different working conditions were:0.009 1, 0.015 1, 0.025 6 MPa, and the maximum displacement values were:2.581 6, 4.259 6, 7.228 4 mm. The maximum stress values and the maximum displacement values were all increased with increasing of abdominal pressure in the two groups. The maximum stress values and the maximum displacement values of the POP patients were greater than those of volunteers. Under different working conditions, the maximum stress values and maximum displacement values of the posterior uterus POP patient were all greater than those of the anterior uterus POP patient. Conclusions The finite element model of USL and CL is completely based on the MRI technology and the model is real and reliable. The increase of abdominal pressure will produce a larger stress and deformation of USL and CL, which is one of the reasons causing the injury of the ligament.

OBJECTIVETo study the relationship of tumor necrosis factor-alpha (TNF-α)-308G/A gene polymorphisms with Henoch-Schonlein purpura nephritis (HSPN) in children.

METHODSUsing the direct DNA sequencing method, polymorphisms in the TNF-α promoter region (-308) were genotyped in 110 Han children with Henoch-Schonlein purpura (HSP group), including 52 children with nephritis and 58 children without nephritis. Plasma TNF-α levels were measured using ELISA. Ninety ethnically matched healthy children were used as the control group.

RESULTSThere were no significant differences in the polymorphisms of TNF-α (-308G/A) between the HSP and control groups (P>0.05). The GA genotype (29% vs 10%) and A allele frequency (18% vs 7%) in HSP children with nephritis (HSPN) were more common than in those without nephritis (P<0.05). Plasma TNF-α levels in HSPN children with GA+AA genotype (7.1±2.3 pg/mL) were significantly higher than those with GG genotype (5.7±1.5 pg/mL) (P<0.05).

CONCLUSIONSTNF-α-308GA genotype and A allele may contribute to the increased risk for the development of nephritis in children with HSP.

Adolescent ; Child ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Genetic ; Purpura, Schoenlein-Henoch ; genetics ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo identify the genetic defect of inherited coagulation factor (F) deficiency in a Chinese family and to explore its molecular mechanism.

METHODSThe activity and antigen of plasma F were measured by photometric test and enzyme-linked immunosorbent assay, and rocket-electrophoresis, respectively. All the exons and exon-intron boundaries of the FA subunit gene were amplified by PCR and then DNA sequencing was performed. Restriction endonuclease analysis was used for the PCR products of the family members and 80 healthy donors to exclude gene polymorphism.

RESULTSRapid dissolution of the proband's fibrin clot occurred within 30 minutes, and antigen of his plasma F was significantly decreased, two compound heterozygous missense mutations (a C to T transition at nucleotide 177,246 which caused Arg703Trp, and a A to G transition at nucleotide 177,286 which caused His716Arg) in exon 15 of FA subunit gene were found. The possibility of gene polymorphism was excluded by restriction endonuclease analysing. Each of these two missense mutations was respectively found in his mother and father. Molecular modeling based on 3D crystallographic data predicted that the mutant protein decreased stability and was likely to be rapidly degraded.

CONCLUSIONSThe inherited F deficiency in the Chinese family is caused by two compound heterozygous missense mutations-Arg703Trp and His716Arg in the FA subunit, which to our knowledge, are reported for the first time.

Base Sequence ; Child ; Exons ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Heterozygote ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; Pedigree

OBJECTIVEThe oxidation of benzo (a) pyrene mediated by 5-lipoxygenase (5-LOX) were investigated in HBE cells in order to provide further proof that lipoxygenase is the alternative pathway for the oxidation of xenobiotics.

METHODSEnzymic experiment: Soybean lipoxygenase (SLO), substrate (benzo[a] pyrene) and other component react in the enzymic system and the reaction product are detected by spectrophotometry. At the same time, in vitro detect of benzo (a) pyrene-DNA adducts with a UV spectrophotometer and HPLC. Cellular experiment: After HBE cells exposure to different poison (B[a]P 4, 8, 16, 32, 64, 128µmol/L, AA-861, naproxen or α- naphthoflavone 0.1, 1, 10 µmol/L) for 24 hours, the effect of benzo (a) -pyrene on cell survival rate were assessed by reductions of tetrazolium dye (MTT) and flow cytometry in cultured HBE cells, and the protein expressions of 5-lipoxygenase in the cells are tested by western-blot, and the DNA damages by the single cell gel electrophoresis. And then, the effect of the specific inhibitor of 5-lipoxygenase (AA-861) on 5-lipoxygenase protein expression and DNA damage in the cells are detected.

RESULTSSLO can catalyze the co-oxidation of benzo (a) pyrene to generate benzo (a) pyrene-7,8-epoxide in the presence of hydrogen peroxide. GTP can inhibit the reaction , the IC50 value is 0.46 mg/L, the model equation is Probit (P) = 0.8985+2.6824 Log (dose). SLO can catalyze the co-oxidation of benzo (a) pyrene to generate a new product, but fail to form DNA adducts in vitro. HBE cell viability decreased with the benzo (a) pyrene concentration increased , but AA-861 and naproxen can inhibit it. Flow cytometry and single cell gel electrophoresis experiments show, Benzo (a) pyrene can induce 5-lipoxygenase protein expression, but AA-861 cannot in HBE. Benzo (a) pyrene causes toxic action and DNA damage in HBE, which can significantly inhibit by AA-861, the difference is statistically significant (P < 0.05).

CONCLUSIONSThe co-oxidate of benzo (a) pyrene by 5-LOX turns into electrophiles that covalently bind to DNA and induce DNA damage, which can be significantly inhibited by AA-861.

Benzo(a)pyrene ; metabolism ; Cells, Cultured ; DNA Adducts ; metabolism ; DNA Damage ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Lipoxygenase ; pharmacology