The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.
Astragalus membranaceus ; chemistry ; Cells, Cultured ; Cytochrome P-450 CYP2E1 ; metabolism ; Humans ; Isoflavones ; pharmacology ; Liver ; drug effects ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Saponins ; pharmacology ; Triterpenes ; pharmacology

Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA (siRNA), the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.

OBJECTIVETo study the protective effect of astragaloside IV on oxidative damages of Chang Liver cells induced by ethanol and H2O2.

METHODThe alcoholic and nonalcoholic oxidative damage models were established on Chang Liver cells with ethanol and H2O2, respectively. The cells viabilities were detected by MTT assay, transaminase activity and antioxidant ability were detected by micro plate and colorimetric method, reactive oxide species (ROS) was detected by DCFH-DA fluorescent probe and cell cycle was detected by flow cytometry. DNA ladder method was used to detect apoptosis.

RESULTBoth kinds of oxidative damage could decrease the viability and antioxidant enzyme activity of Chang Liver cells, and increase the transaminase activity and MDA content of extracellular fluid. The protective effects of astragaloside IV against those two kinds of oxidative damages were significant or extremely significant. Meanwhile, ethanol could decline the level of ROS significantly in the damaged cells, while H2O2 could increase it significantly. And the effect of astragaloside IV was to make ROS return to the normal level. Retardation of cell cycle progression of Chang Liver cells in G0/G1 induced by ethanol or H2O2 was relieved, and apoptosis was also inhibited.

CONCLUSIONAstragaloside IV had protective effect on oxidative damages of Chang Liver cells induced by ethanol and H2O2.

Antioxidants ; metabolism ; Apoptosis ; Cell Cycle ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Ethanol ; pharmacology ; Humans ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Saponins ; pharmacology ; Triterpenes ; pharmacology

OBJECTIVETo study the validity and reliability of the Demirjian distinguish software on dental age estimation.

METHODThe dental age of 60 children were estimated with both the software three times and manual measures.

RESULTSThe correlation coefficients of dental age estimated by the software and by manual measure was 0.974 (female) (P > 0.1) and 0.970 (male) (P > 0.05); the coefficients of interclass correlation of each dental age estimated by the software was 0.977 (female) (P > 0.1) and 0.977 (male) (P > 0.05).

CONCLUSIONDemirjian distinguish software has high validity and reliability in estimating dental age.

Adolescent ; Age Determination by Teeth ; methods ; Child ; Female ; Humans ; Male ; Reproducibility of Results ; Software

OBJECTIVETo investigate the application of the multiple locus variable numbers of tandem repeats (MLVA) in genotype Mycobacterium tuberculosis strains isolated from Tibet, and to understand the characteristics of genotype and distribution.

METHODS217 M. tuberculosis strains were collected from six regions of Tibet. Twenty tandem repeats loci in the total genome of M. tuberculosis (MTB) were analyzed by PCR and agarose gel electrophoresis method. The characteristics on polymorphism of DNA fingerprinting of 217 MTB strains were analyzed with BioNumerics 3.0 software.

RESULTS217 M. tuberculosis strains detected with 20 MLVA loci were classified to 19 genotypes with 87.6% of the stains belonging to Beijing genotype and the other 18 genotypes were scattered,accounted for 1.38% and 0.92% strains, respectively. Beijing genotype was not significantly associated with the resistance to all of the four drugs and BCG vaccination.

CONCLUSIONIt is concluded that the strains of MTB isolated in Tibet present definite polymorphism and most of the epidemic strains belonged to Beijing family genotype and MTB genotyping. The Beijing genotype was not recognized as the one transferred from some of the drug resistance strains or from BCG vaccination. Being a fast and simple technique, MLVA method, seemed a better molecular typing method and could be used for genotyping in M. tuberculosis and monitoring pathogen.

BCG Vaccine ; DNA Fingerprinting ; Electrophoresis, Agar Gel ; Genotype ; Minisatellite Repeats ; genetics ; Mycobacterium tuberculosis ; classification ; genetics ; Phylogeny ; Polymerase Chain Reaction ; Tibet

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Glycine ; analogs & derivatives ; blood ; Humans ; Inosine Monophosphate ; blood ; Reproducibility of Results ; Sulfonamides ; blood ; Tandem Mass Spectrometry

OBJECTIVETo study the weak loop in the treatment of children with out-of-hospital cardiac arrest (OHCA) and the treatment strategy for improvement.

METHODData of a total of 133 patients with OHCA who were rescued by Wuxi pre-hospital care center during the 2005 - 2011 were analyzed.

RESULTThe main causes of pediatric OHCA were drowning (52/133) and accidental injury disease (30/133). The cases of OHCA were mainly located in public places (60/133) and the majority occurred in winter and summer. The emergency rescue response time was (13.21 ± 8.09) min, the rate of first witness treatment was 3.91%, intubations was performed in 11.11%, opening of vein access was 23.15% and one case got restoration of spontaneous circulation (ROSC).

CONCLUSIONThe rate of ROSC of pre-hospital cardiac arrest in children was significantly lower than that of hospital cardiac arrest. Preventive interventions on children's accidents and the skills of pre-hospital staff on pediatric advanced life support (PALS) need to be urgently improved.

Advanced Cardiac Life Support ; education ; Cardiopulmonary Resuscitation ; methods ; Chi-Square Distribution ; Child ; Child, Preschool ; China ; epidemiology ; Emergency Medical Services ; methods ; organization & administration ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Out-of-Hospital Cardiac Arrest ; epidemiology ; etiology ; therapy ; Pediatrics ; education ; Retrospective Studies ; Risk Factors ; Survival Rate ; Time Factors ; Treatment Outcome

OBJECTIVETo study the clinical effect and feasibility of blood type A donor liver transplantation in blood type O recipients.

METHODSThe clinical data were analyzed in 6 blood type O patients receiving transplantation of the liver grafts from blood type A donors. The clinical effect and outcomes of the transplantations were evaluated to assess the feasibility of ABO incompatible liver transplantation between type A donors and type O recipients.

RESULTSThe operations and the postoperative recovery were smooth in all the 6 recipients. Only one patient died 5 months postoperatively due to liver tumor metastasis, and the other 5 patients survived with the longest survival reaching 14 months. Acute graft rejection occurred in one patient 1 week after the operation on account of abnormally elevated serum bilirubin level, which was successfully managed with immediate methylprednisolone therapy. No such complications as acute graft rejection, bile duct stenosis or bile leakage was found in the other patients.

CONCLUSIONBlood type A donor liver transplantation in type O recipient is feasible in emergency or other special conditions.

ABO Blood-Group System ; immunology ; Adult ; Blood Group Incompatibility ; immunology ; Female ; Graft Survival ; Humans ; Liver Transplantation ; immunology ; Male ; Middle Aged ; Retrospective Studies ; Tissue Donors

OBJECTIVETo investigate the impact of hepatic steatosis on virologic response in chronic hepatitis B (CHB) patients treated with pegylated interferon-alpha (PEG-IFNa).

METHODSNinety-six naive patients positive for hepatitis B e antigen (HBeAg) and with biopsy-proven CHB were administered PEG-IFNa-2a or PEG-IFNa-2b for 48 weeks. Virologic response (HBeAg clearance and hepatitis B virus (HBV) DNA less than 5 log10 copies/ml) and biochemical response (alanine transaminase (ALT) normalization) were compared between patients with (n=34) and without (n=62) steatosis.

RESULTSThe HBV DNA titer in the steatosis group was significantly lower than that of the non-steatosis group (6.961.27 vs. 7.541.28 log10 copies/ml; t=2.161, P=0.033). After 48 weeks of PEG-IFNa treatments, there was no significant difference in HBeAg seroconversion or the percentage of undetectable HBV DNA (less than 3 log10 copies/ml) between steatosis and non-steatosis patients. However, the steatosis patients presented with a significantly lower complete response rate (virologic response plus biochemical response) compared to non-steatosis patients (26.5% vs. 48.4%; x² =4.373, P=0.037). Of the 45 CHB patients with undetectable HBV DNA after 48 weeks of treatment, seven did not achieve ALT normalization. The rate of patients with non-biochemical response was significantly higher in the steatosis group than in the non-steatosis group (33.3% vs. 6.67%; P=0.032).

CONCLUSIONHepatic steatosis does not affect the virologic response, but does affect the biochemical response in CHB patients treated with PEG-IFNa for 48 weeks.

Adult ; Antiviral Agents ; therapeutic use ; Fatty Liver ; complications ; pathology ; virology ; Female ; Hepatitis B, Chronic ; complications ; drug therapy ; pathology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Young Adult

PURPOSE: To compare and assess the efficacy, safety and utility of hand-assisted laparoscopic surgery (HALS) with open surgery (OS) in total colectomy with ileorectal for colonic inertia. METHODS: From January 2001 to February 2012, 56 patients diagnosed with colonic inertia who failed to respond to medical treatments underwent hand-assisted laparoscopic total colectomy with ileorectal anastomosis. Another 68 patients underwent laparotomy. Main parameters such as clinical manifestations, conversion to open procedure, operative time, incision length, pain score, intraoperative blood loss, time to first flatus and hospitalization, early postoperative complications and hospitalization cost were retrospectively analyzed. Postoperative defecating frequencies were followed up in both groups. RESULTS: All patients received successful operation, no surgical mortality happened and none of the patients required conversion to an exploratory laparotomy in HALS group. The clinical features, the estimated blood loss, incision length, pain score, first passing flatus time, and postoperative hospitalization time were superior in HALS group (P < 0.05). The early postoperative complications and frequency of defecation were similar. However, the mean operative time was longer and hospitalization cost was higher in HALS group than those in OS group (P < 0.05). CONCLUSION: HALS total colectomy can be a safe and efficient technique in the treatment of colonic inertia. HALS can result in a better cosmetic result and a quicker postoperative recovery, but requires higher direct cost.
Colectomy ; Colon ; Constipation ; Cosmetics ; Defecation ; Flatulence ; Hand-Assisted Laparoscopy ; Hospitalization ; Humans ; Laparotomy ; Operative Time ; Postoperative Complications ; Retrospective Studies ; Surgical Procedures, Operative