Objective To investigate the reference values of urinary formed elements detected by Sysmex UF‐1000i fully auto‐matic urine analyzer (UF‐1000i) in healthy people with different genders and different ages in Lanzhou .Methods 476 cases of clean midway urine samples were collected from healthy people .UF‐1000i was used to detect the urinary formed elements ,including WBC ,RBC ,EC ,CAST and BACT .Results There were differences in the results of urinary formed elements among healthy people with different genders and different ages .And the reference values of WBC ,RBC ,EC ,CAST and BACT were established respective‐ly in the crowds of male and female children ,adults ,and elderly .Conclusion It is necessary to establish the reference values of uri‐nary formed elements for the crowds with different genders and different ages in some region .

Objective In order to verify the validation of the industry standard WS/T 405‐2012 about adult blood cell analysis reference range ,and ensuring their clinical application .Methods According to Clinical and Laboratory Standards Institute(CLSI) C28‐A3 recommendation method ,40 health reference individuals were enrolled in the study whose blood samples were collected ,de‐tected and analyzed .Results The blood cell analysis indicators involved in the validation included WBC ,RBC ,hemoglobin(HGB) , hematocrit(HCT) ,mean corpuscular volume(MCV) ,mean corpuscular hemoglobin(MCH) ,mean corpuscular hemoglobin concen‐tration(MCHC) ,platelet(PLT ) ,etc .The probability of all results of selected 40 reference individuals beyond the WS/T 405‐2012 blood cell analysis reference interval were no more than 10% (value 90% or higher) .Conclusion The WS/T 405‐2012 blood cell a‐nalysis reference range is suitable for the laboratory .A perfective verification system should be established to ensure its application .

Objective To explore the changes of programmed death receptor-1 (PD-1) in chronic hepatitis B (CHB) patients with different baseline of hepatitis B virus (HBV) DNA levels after treatment with adefovir dipivoxil (ADV).Methods One hundred CHB patients with positive hepatitis B e antigen (HBeAg),1 × 104 copy/mL≤HBV DNA≤1 × 107 copy/mL,and positive human leukocyte antigen-A2 were divided into two groups according to the baseline HBV DNA level:47 cases in low virus load group whose HBV DNA level was ≤1 × 105 copy/mL; 53 cases in high virus load group whose HBV DNA level was＞1 × 105 copy/mL.Both groups were treated with ADV 10 mg/d.Serum HBV DNA,HBeAg seroconversion rate,alanine aminotransferase (ALT) and total bilirubin (TBil) levels of both groups before treatment and 12 months after treatment were compared.Flow cytometry was used to test peripheral blood HBV-specific cytotoxic T lymphocyte (CTL) surface PD-1 and peripheral blood HBV-specific CTL level.Categorical data were tested by x2 test; quantitative data was compared with t-test.Results Peripheral blood HBV-specific CTL surface PD-1 of CHB patients in low virus load group was 20.17 ％±1.69％,which was lower than that in high virus load group (41.38％±2.30％,t =53.02,P＜0.01) ; peripheral blood HBV specific CTL levels in two groups were 0.37％±0.02％ and 0.17％± 0.02％,respectively (t=50.47,P＜0.01) ; ALT and TBil levels in low virus load group were both lower than those of high virus load group (t=13.07,P＜0.01; t=5.06,P＜0.01).Twelve months after treatment,HBV DNA of 25 cases (53.2％) in low virus load group and 10 cases (18.9％) in high virus load group were lower than the detectable level (HBV DNA＜500 copy/mL,x2 =12.89,P＜0.01);HBeAg seroconversion was achieved in 15 cases(31.9％) and 1 case (1.9％),respectively (x2 =16.72,P＜0.01) ; peripheral blood HBV-specific CTL surface PD-1 expression levels were 9.00 ％ ±1.38 ％ and 29.40 ％ ± 3.76 ％,respectively (t =36.80,P＜ 0.01) ; peripheral blood HBV-specific CTL levels were 0.65％±0.10％ and0.48％±0.07％,respectively (t=9.61,P＜0.01).Conclusions After treatment with ADV,along with the decrease of HBV DNA load,HBV-specific CTL surface PD-1 expression decreases,while HBV-specific CTL level increases.The changes in low virus load group are much more remarkable.

Objective To explore the possible association of single nucleotide polymorphism (SNP) rs3738435 of muscarinic acetylcholine receptor subtype M3 gene (cholinergic receptor, muscarinic 3, CHRM3) with risk of type 2 diabetes mellitus (DM) and metabolic disturbance. Methods The genotypes of T-149C variant of CHRM3 gene were determined by PCR-RFLP in 573 Chinese individuals in Shanghai, including 220 newly-diagnosed type 2 DM patients without taking any drug and 353 subjects with normal glucose tolerance (NGT). In the subjects, height and weight were measured for body mass index(BMI), waist, hip and femoral circumstances for waist-to-hip ratio (WHR) and waist-to-femur ratio (WFR), and serum lipid level including total cholesterol, triglyceride, high-density and low-density lipoprotein cholesterol, blood pressure, plasma glucose levels both at 0 and 120 minute during oral 75 g glucose tolerance test (OGTT) were also determined. Results (1) There was no statistical difference in the gene frequency between groups of type 2 DM and NGT. (2) In the group of type 2 DM, significant differences were observed between TT genotype carriers and TC+CC genotypes carriers for BMI, with an obvious increase in TY genotype carriers [(26.99±3.59vs25.34±3.48)kg/m2, P=0.001]. (3) In the subgroup of type 2 DM with BMI≥25 kg/m2, total cholesterol was higher in TT genotypes than in TC+CC genotypes[(5.75±1.26vs5.27±1.14)mmol/L, P=0.030], so was the low-density lipoprotein cholesterol. Conclusion The genetic variation T-149C in the CHRM3 gene seems to attribute to weight regulation and lipid metabolism of patients with type 2 diabetes mellitus in Chinese population.

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism. Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats. A total of 30 SD male adult rats were randomly divided into 5 groups: group A (sham operation, n=6); group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery, n=6); group C (in which the rats received 30 min of ischemia, n=6), group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min, n=6 in each). The left ventricular tissues were removed immediately after the ischemia-reperfusion injury. Neonatal cardiomyocytes were cultured and treated with different concentrations of H(2)O(2) (0, 5, 10, 20 μmol/L) or tumor necrosis factor-α (TNF-α, 0, 1, 5, 10 ng/mL). The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR, western blotting and immunohistochemistry. Additionally, neonatal cardiomyocytes were transfected with Pim-3 siRNA, and induced to develop apoptosis by using H(2)O(2). The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein. Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues. Furthermore, H(2)O(2) but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes. And Pim-3 silencing failed to strengthen the H(2)O(2)-inducing apoptosis in cardiomyocytes. It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.

Pim kinases contribute to tumor formation and development of lymphoma, which shows enhanced DNA replication, DNA recombination and repair. Endothelial cells^(ECs) express all the three members of Pim kinase gene family. We hypothesized that DNA repair gene would regulate Pim expression in ECs. Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium. The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining. The siRNA fragments were synthesized and transfected by using Lipofectamine LTX. The total cellular RNA was extracted from the cells by using Trizol reagent. cDNAs were quantified by semi-quantity PCR. The effects of LY294002 and wortmannin on RNA stability in ECs were also examined. Our data showed that LY294002 and wortmannin, phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors, increased Pim mRNA expression in ECs without altering the mRNA stability. RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1, respectively. Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs. But etoposide, a nucleoside analogue, which could activate DNA-PKcs and ATM, increased Pim expression in ECs. Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism. Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats. A total of 30 SD male adult rats were randomly divided into 5 groups: group A (sham operation, n=6); group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery, n=6); group C (in which the rats received 30 min of ischemia, n=6), group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min, n=6 in each). The left ventricular tissues were removed immediately after the ischemia-reperfusion injury. Neonatal cardiomyocytes were cultured and treated with different concentrations of H(2)O(2) (0, 5, 10, 20 μmol/L) or tumor necrosis factor-α (TNF-α, 0, 1, 5, 10 ng/mL). The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR, western blotting and immunohistochemistry. Additionally, neonatal cardiomyocytes were transfected with Pim-3 siRNA, and induced to develop apoptosis by using H(2)O(2). The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein. Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues. Furthermore, H(2)O(2) but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes. And Pim-3 silencing failed to strengthen the H(2)O(2)-inducing apoptosis in cardiomyocytes. It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.
Animals ; Animals, Newborn ; Male ; Myocardial Ischemia ; metabolism ; physiopathology ; Myocardial Reperfusion Injury ; metabolism ; Myocardium ; metabolism ; Oxidative Stress ; physiology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Up-Regulation

Objective To study the prevalence of common infections with soil-borne intestinal nematodes amongst kindergarten children aged 3 to 6 years in Hangzhou,Zhejiang Province to provide evidence for determination of the priority of disease prevention and control.Methods Totally,1667 preschool children were selected from 14 kindergartens of Classes A,B and C in east,middle and west Hangzhou.Perianal skin Scotch Tape(a short strip of sealing cellophane pressure-sensitive tape)specimens were collected for detection of eggs of Enterobius vermicularis,and stool specimens for eggs of Ascaris lumbricoides,Ancylostoma duodenale and Trichuris trichiura by Kato-Katz method and saturated brine floatation,as well as questionnaire interview,for all the children.Results Two hundred and sixteen of 1667 children examined were found infected with common soil-borne intestinal nematodes,with an overall prevalence of 12.96%,4.44% for Enterobius vermicularis,8.28% for Ascaris lumbricoides,0.54% for Trichuris trichiura and 0.24% for Ancylostoma duodenale.Prevalence of infection of common intestinal nematodes was 7.31% in children of the Class A kindergartens,12.60% of Class B,and 21.47% of Class C,with statistically significant difference(?~2 = 49.95,P

Pim kinases contribute to tumor formation and development of lymphoma,which shows enhanced DNA replication,DNA recombination and repair.Endothelial cells (ECs) express all the three members of Pim kinase gene family.We hypothesized that DNA repair gene would regulate Pim expression in ECs.Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium.The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining.The siRNA fragments were synthesized and transfected by using Lipofectamine LTX.The total cellular RNA was extracted from the cells by using Trizol reagent.cDNAs were quantified by semi-quantity PCR.The effects of LY294002 and wortmannin on RNA stability in ECs were also examined.Our data showed that LY294002 and wortmannin,phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors,increased Pim mRNA expression in ECs without altering the mRNA stability.RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1,respectively.Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs.But etoposide,a nucleoside analogue,which could activate DNA-PKcs and ATM,increased Pim expression in ECs.Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.