OBJECTIVETo study DNA damages of liver cells in rats exposed to vinyl chloride monomer (VCM), and the expressions of DNA damage repair enzymes including O(6)-methyl guanine-DNA methyl transferase (MGMT), X-ray repair cross-complementing group 1 (XRCC1) and X-ray repair cross-complementing group 3 (XRCC3); and to explore the repair mechanism of DNA damage induced by VCM.

METHODSRats were exposed to VCM by intraperitoneal injection. DNA damages were detected by single cell gel electrophoresis (comet assay). The expressions of DNA damage repair enzymes were measured by immunohistochemical methods.

RESULTSThe percentages of comet cells in low, moderate, and high dose groups (11.75%, 12.38%, and 17.63%, respectively) were greater than that of control (5.67%). The latter two groups were significantly different from that of control (P < 0.05, P < 0.01). The expressions of MGMT and XRCC1 decreased, and XRCC3 increased with the dose of VCM increased. DNA damage was correlated with the expression of XRCC3 (r = 0.438, P = 0.067).

CONCLUSIONVCM can cause DNA damage of liver cells with dose-response relationship. DNA damage repair enzymes take part in the repairing of DNA damage induced by VCM.

Animals ; Carcinogens ; toxicity ; DNA Damage ; drug effects ; DNA Repair ; DNA-Binding Proteins ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Liver ; cytology ; metabolism ; Male ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vinyl Chloride ; toxicity ; X-ray Repair Cross Complementing Protein 1

OBJECTIVETo investigate the relationship between the thiopurine methytransferase (TPMT) gene polymorphisms and its enzymatic activity, and to clarify the significance of TPMT activity and gene polymorphisms on individualized therapy with thiopurines.

METHODSThe TPMT activity and gene polymorphisms were determined in an unrelated population of 250 Chinese healthy blood donors, 100 cords blood and 280 patients with acute leukemia. The TPMT genotyping assay was based on polymerase chain reaction (PCR), restriction digestion of PCR products, denaturing high-performance liquid chromatography (DHPLC) and SNaPshot sequencing and direct DNA sequencing in the TPMT exon 5 (G238C), TPMT exon7 (G460A) and TPMTexon10 (A719G). Erythrocyte TPMT activity was measured by high-performance liquid chromatography (HPLC).

RESULTSThe frequency of TPMT polymorphism in 250 Chinese healthy blood donors, 100 cords blood and 280 patients with acute leukemia was low (3.5%), and all the varied alleles were TPMT* 3C (exon 10A719G). All of them were TPMT* 1/TPMT* 3C heterozygote. The TPMT activity was between 6 and 12 U. The activity in 95.1% was more than 12 U (13 - 32 U), while the activity in others (4.9%) was 6 - 12 U. TPMT activity and genotype were concordant. Of 630 subjects evaluated, TPMT activity of heterozygous individuals in Chinese healthy blood donors, cords blood and acute leukemia patients were 9.1 U, 9.3 U and 9.07 U, respectively, significantly lower than that in general population (17.6 U, 17.67 U and 18.6 U, respectively). In the samples analyzed, ten subjects with heterozygous phenotypes (6/15 acute leukemia children and 4/16 healthy blood donors and cords blood) did not have TPMT* 2, TPMT* 3A or TPMT* 3C. Therefore, other factors may affect on TPMT activity.

CONCLUSIONTPMT gene polymorphisms and its activity were concordant. The heterozygotes had low TPMT activity. Therefore, detection of TPMT genotype and its activity is useful. These findings hold a promise of improving the safety and efficacy of thiopurines therapy.

Acute Disease ; Child ; Chromatography, High Pressure Liquid ; Erythrocytes ; enzymology ; Exons ; Female ; Fetal Blood ; enzymology ; Genotype ; Humans ; Leukemia ; blood ; enzymology ; genetics ; Male ; Methyltransferases ; blood ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo explore the associated risk factors of metabolic syndrome in Jiangsu province, China.

METHODSUsing identical protocol and questionnaire, an epidemiological study was carried out in a population of 5888 adults in 12 counties in Jiangsu. Anthropometric test and blood sampling were conducted at the time of interview. IDF (2005) was used as the diagnostic criteria of metabolic syndrome . The prevalence and age-standardized prevalence of metabolic syndrome were calculated. Univariate and multivariate logistic regression model were used to identify associated risk factors.

RESULTSThe prevalence and age-standardized prevalence of metabolic syndrome in Jiangsu were 19.07% (11.10% in males and 25.72% in females) and 17.48% (11.49% in males, 22.86 % in females), respectively. Among the potential risk factors of metabolic syndrome as gender, age, education level, occupation, income, physical activity, smoking, alcohol drinking, disease family history, data from univariate and multivariate logistic regression analyses suggested that gender (OR = 1.91), age (OR = 1.15), physical inactivity (OR = 1.94), with hypertension family history (OR = 1.99) and with obesity family history (OR = 6.24) could significantly increase the risk of disease development.

CONCLUSIONMetabolic syndrome has become a significant public health problem among the adults in Jiangsu province.

Adult ; Age Factors ; Aged ; Alcohol Drinking ; adverse effects ; China ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; blood ; epidemiology ; genetics ; Middle Aged ; Motor Activity ; physiology ; Risk Factors ; Sex Factors ; Smoking ; adverse effects ; Surveys and Questionnaires

OBJECTIVETo study the clinical and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) infection in children.

METHODA total of 37 MRSA strains were isolated from hospitalized patients in Children's Hospital of Fudan University from March 2009 to November 2011. The clinical characteristics were investigated by a cohort study. Furthermore, the mecA, Panton-Valentine leucocidin (PVL) genes were detected by polymerase chain reaction (PCR), and the genotypes of SCCmec were determined by multiplex PCR.

RESULT(1) Among the 37 MRSA isolates, infections with 21 were acquired from hospital (HA-MRSA), and 16 isolates were acquired from community (CA-MRSA). (2) In the study, MRSA frequently caused respiratory tract infection, and most of the strains were isolated from intensive care unit (ICU). (3) CA-MRSA was most frequently associated with skin and soft tissue infections (SSTI), suppurative tonsillitis, even pneumonia and septicemia. HA-MRSA infection was more aggressive, most frequently associated with pneumonia, septicemia, and central nervous system (CNS) infections, such as meningitis. In children with fever caused by HA-MRSA or CA-MRSA infection, HA-MRSA showed a longer duration of fever, for 10.5 days. C-reactive protein (CRP) level caused by HA-MRSA (63.00 mg/L) was higher than CA-MRSA (9.50 mg/L) , and there were statistically significant differences between the groups (t = 2.5670, P < 0.05). However, there were no statistically significant differences between the groups in white blood cell count (WBC) or procalcitonin (PCT) level. (4) Among 37 MRSA isolates, the whole isolates were mecA gene positive (100%). SCCmec genotyping results showed that the most frequent SCCmec types were type III, 17 isolates, the others including type IV 8 isolates, type II1 isolates, nontypable 11 isolates, type I and type V were not found in this group. Therein, among 21 HA-MRSA isolates, SCCmec III was the most common, 15 isolates, type IV 1 isolates, nontypable 5 isolates; among 16 CA-MRSA isolates, SCCmec type IV was the most common, 7 isolates, type III 2 isolates, type II 1 isolate, nontypable 6 isolates. (5) Among the 37 MRSA isolates, 28 were PVL gene positive; and among 21 HA-MRSA isolates, 17 were PVL gene positive; Among 16 CA-MRSA isolates, 11 were PVL gene positive; There were no statistically significant differences between the groups (χ(2) = 0.735, P > 0.05) .

CONCLUSIONCompared with CA-MRSA, HA-MRSA infection was more aggressive, and induced higher C reactive protein; the dominant epidemic strains of CA-MRSA was SCCmec type IV, and HA-MRSA was SCCmec type III; the positive rate of PVL gene was high.

Adolescent ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; Bacterial Typing Techniques ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Community-Acquired Infections ; epidemiology ; microbiology ; Cross Infection ; epidemiology ; microbiology ; DNA, Bacterial ; genetics ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Methicillin ; pharmacology ; Methicillin Resistance ; genetics ; Methicillin-Resistant Staphylococcus aureus ; classification ; genetics ; isolation & purification ; Penicillin-Binding Proteins ; Staphylococcal Infections ; epidemiology ; microbiology

The aim of the present study was to gain an insight into the thiopurine methytransferase (TPMT) genotyping assay, which was based on polymerase chain reaction (PCR), allele-specific PCR, restriction digestion of PCR products, denaturing high-performance liquid chromatography (DHPLC) and SNaPshot sequencing and in combination with direct DNA sequencing. Among the f our methods to test TPMT genetic SNPs based on PCR, allele specific PCR was not able to differentiate wild type from varied type. BsiYI, MwoI and AccI to digest PCR products were used so that SNP in TPMT exon 5, 7 and 10 tested. It showed that there were no differences between the results of digestion of PCR products and those of DNA sequence analysis. Therefore, this method was reliable. But some other methods were still needed to look for a compensation, because no restriction map changing resulted from the 2 SNPs in TPMT promotor was found. As to the results of DHPLC, those for the screening of TPMT exon-5 and -10 for SNPs were the same as restriction analysis of PCR products and direct DNA sequencing. But the variation of the heterozygotes in exon-7 was high, which was different from the results of direct DNA sequencing. After changing the Tm of DNA step by step, It was found that all the samples showed single peak when the temperature was 54 degrees C. But this result was unbelievable because a heterozygote in exon 7 as positive control could not be found. Therefore, it was necessary to test the sensitivity and accuracy of DHPLC, though DHPLC could be used as an effective method of SNPs screening. The results of the SNaPshot sequencing were also same as those of restriction analysis of PCR products and direct DNA sequencing. And the results showed that the bases of TPMT promoter -91 and -168 were G, instead of A and T. The results of the four methods to detect TPMT genetic SNPs based on PCR showed that SNPs analysis technique should be a combination of the techniques above-mentioned. One technique alone could not satisfy the need in clinics and research. The compensation of each other was very important.
Chromatography, High Pressure Liquid ; Exons ; Humans ; Methyltransferases ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the presence of pathogenicity island (PAI)-associated genes in the enterococcal isolates.

METHODSUsing PCR and hybridization methods, PAI-associated genes were detected in 155 enteococcal strains isolated from clinical patients and healthy individuals.

RESULTSAmong the 155 enterococcal isolates, 137 (88.39%) carried at least one of PAI-associated genes, namely hyd (positivity rate of 81.94%), psaA (78.06%), nuc (57.42%), esp (53.55%), cylB (52.90%), and gls24-like (38.06%) genes. Expect for esp gene, the other 5 genes showed higher positivity rates in the E. faecalis strains than in the E. faecium strains, and this difference was statistically significant for the genes nuc, cylB, and gls24-like. The positivity rates and the number of these genes in the E. faecalis from clinical isolates were both significantly higher than those in the strains isolated from healthy individuals.

CONCLUSIONThe data show a wide distribution of the PAI-associated genes among the enterococcal strains, and E. faecalis strains are more likely than E. faecium strains to be positive for the 6 genes, which are present at significant higher rates in the clinically isolated samples than in that from healthy individuals.

Bacterial Proteins ; chemistry ; genetics ; Enterococcus ; genetics ; isolation & purification ; pathogenicity ; Enterococcus faecalis ; genetics ; isolation & purification ; pathogenicity ; Genomic Islands ; genetics ; Gram-Positive Bacterial Infections ; microbiology ; Humans ; Membrane Proteins ; genetics ; Virulence ; genetics

AIM To prepare nanosuspensions of flavonoids from Glycyrrhizae Radix et Rhizoma and to determine the in vitro dissolution rate.METHODS Precipitation-high pressure homogenization method was adopted in the preparation of nanosuspensions.With mean particle size and polydispersity index (PDI) as evaluation indices,concentrations of flavonoids,povidone K30 (PVP K30) and polyethylene glycol 400 (PEG 400) as influencing factors,central composite design-response surface method was applied to optimizing the preparation.For the freedried powder prepared by freeze-drying method,the optimal kind and ratio of lyoprotectant were screened.Then the in vitro dissolution rates of freeze-dried powder and physical mixture were compared.RESULTS The optimal conditions were determined to be 10.00 mg/mL for flavonoids' concentration,and 2.30 mg/mL for both PVP K30 and PEG 400 concentrations,the mean particle size and PDI were (172.3 ± 1.2) nm and 0.175 ± 0.004,respectively.The optimal lyoprotectant was 5％ mannitol-lactose (3 ∶ 2),the mean particle size and PDI after redissolution were (239.7 ±2.1) nm and 0.193 ±0.032,respectively.The in vitro dissolution rate of lyoprotectant reached 87.7％ within 60 min,which was much higher than that of physical mixture (less than 30％).CONCLUSION Nanosuspension can effectively improve the in vitro dissolution rate of flavonoids from Glycyrrhizae Radix et Rhizoma.

Adolescent ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; therapy ; Humans ; Male ; Middle Aged ; Plasma Exchange ; methods ; Severity of Illness Index ; Sorption Detoxification ; methods ; Treatment Outcome

OBJECTIVESOrthotopic liver transplantation (OLT) is an accepted therapy for selected patients with advanced liver diseases. However, the early mortality rate after OLT remains relatively high due to the poor selection of candidates with various serious conditions. The aim of this study is to assess the value of pretransplantation artificial liver support treatment in reducing the pre-operation risk factors relating to early mortality after OLT.

METHODS50 adult patients in various stages of different etiologies who underwent OLT procedures had been treated with molecular adsorbent recycling system (MARS) preoperatively. The study was designed in two parts: the first one was to evaluate the effectiveness of a single MARS therapy by using some clinical and laboratory parameters which were supposed to be therapeutical pretransplantation risk factors. The second part was to study the patients undergoing OLT by using the regression analysis on preoperation risk factors relating to early (within 30 d after OLT) mortality rate.

RESULTSAmong the 50 patients, a statistically significant improvement of the biochemical parameters was observed (pretreatment vs posttreatment). 8 patients cancelled their scheduled LTXs due to significant improvements in their clinical conditions or recovery of their failing liver functions. 8 patients died and 34 patients successfully underwent LTX. The immediate outcome (within 30 postoperative days) of these 34 patients was that 28 were kept alive and 6 died.

CONCLUSIONSPreoperation sequential organ failure assessment (SOFA), level of creatinine, INR, TNFalpha, and IL-10 are the main preoperative risk factors relating to early death after an operation. MARS treatment before a transplant operation can relieve these factors significantly, hence improve survival rate of liver transplantation or even make the transplantation unnecessary.

Aged ; Factor Analysis, Statistical ; Female ; Humans ; Interleukin-10 ; blood ; Liver Cirrhosis ; surgery ; Liver Neoplasms ; surgery ; Liver Transplantation ; methods ; Liver, Artificial ; Male ; Middle Aged ; Preoperative Care ; Risk Factors ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo describe the prevalence of metabolic syndrome (MS) in Jiangsu province, using IDF 2005 and ATP III 2005 definition.

METHODSMultistage cluster sampling method was used and the participants were interviewed by trained health workers under a structured questionnaire. The number of research subjects was 5888. Fasting plasma glucose (FPG), blood lipids, body mass index, blood pressure (BP) of all sample population (5888 subjects) were measured.

RESULTSThe prevalence of MS was 17.48% by IDF 2005 definition and 21.95% by ATP lII 2005 definition. Women had higher prevalence than men (P < 0.01). The prevalence of MS increased with age. Abrupt increase of the prevalence started at age of 50 in women. The prevalence varied by education, marital status, occupation and income. People from the urban and the south had a higher prevalence than from the rural and from the northern part of the country. The most common combination of MS individual components were 1) central obesity, dislipidemia and high BP; 2) central obesity, dislipidemia, high BP and high FPG.

CONCLUSIONThe prevalence of MS was high in Jiangsu province. Clustering of MS components was common. It is necessary to discuss the cut-off points of central obesity for the waist circumference diagnostic criteria of MS in Chinese population. The importance of prevention of MS should be strengthened both by health professionals and the government.

China ; Female ; Humans ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Obesity, Abdominal ; epidemiology ; Prevalence ; Waist Circumference