Objective To construct and express the recombinant plasmid pET28α-Sj26GST-Sj32 of Schistosoma japonicum(Sj) in Escherichia coli BL21 (DE3). Methods Total RNA was extracted from Sj adult worms by ultrasound-breaking, Sj26GST and Sj32 antigen gene was respectively amplified by RT-PCR from the total RNA; Sj26GST-Sj32 fusion gene obtained with gene splicing by overlap extension(SOEing) was cloned into prokaryotic expression plasmid pET28α and transformed into Escherichia coli BL2 (DE3) to construct pET28α-Sj26GST-Sj32;BL21 (pET28α-Sj26GST-Sj32) was induced with isopropyl-β-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)and Western blotting. Results The 1991 bp Sj26GST-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into pET28α by restriction analysis and PCR identification, the recombinant plasmid pET28α-Sj26GST-Sj32 was successfully constructed; the relative molecular mass of the expressed recombinant protein was approximately 69 × 103 by SDS-PAGE, and the amount of the expressed protein was 25% of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pET28α-Sj26GST-Sj32 is successfully constructed and highly expressed in Escherichia coli in fused form with His-tag, and the expressed fusion protein shows specific antigenicity.

Objective To construct and express the recombinant plasmid pET32α-Sj26GST of Schistosoma japonicum(sj)in Escherichia coli(E.coli)B121(DE3).Methods The total RNA was extracted from sj adult worms by ultrasound-breaking,Sj26GST antigen gene was amplified by RT-PCR from the total RNA,then cloned into prokaryotic expression plasmid pET32α(+) and transformed into E.coli B12(DE3)to construct pET32α-Sj26GST;BL21(pET32α-Sj26GST)WaS induced with isopropyl-β-D-thiogalactopyranosid(IPTG),and the expressed products were analyzed and identified by SDS-PAGE and Western blot.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET32α(+)by restriction analysis and PCR identification,the recombinant plasmid pET32α-Sj26GST was successfully constructed;the relative molecular mass of the expressed recombinant protein was approximately 49×103 by SDS-PAGE,and the amount of the expressed protein was 24%of the total bacterial proteins;the fusion protein could be recognized by sera from rabbits infected with sj by Western blot.Conclusions The recombinant plasmid pET32α-Sj26GST is successfully constructed and highly expressed in E.coli in fused form with Trx-tag and His-tag,and the expressed fusion protein shows specific antigenicity.

Objective To prepare a glycerol reference material.Methods The material was prepared and characterized according to the primary standard substance technological specification(JJG 1006- 1994).Glycerol was dissolved in water containing 0.5% sodium azide and dispersed to glass ampules.The homogeneity and stability of this material were tested with an HPLC method.Glycerol concentration was determined by a titration method as specified in the Pharmacopoeia of China.Results The three time measuring result of glycerol reference material was 1.297 5?0.014 3,1.302 0?0.008 9,1.313 7? 0.007 8,respectively.Statistical analysis showed that this material was homogeneous (F=1.462,P=0.166) .It was stable for at least 4 years at 4℃.The assigned reference value was 0.103 6 g/g and the expanded uncertainty was 0.000 4 g/g.Conclusions This material meet the technical requirements of national primary standard reference material.It is approved as the Certified Reference Material (GBW 09149) by General Administration of Quality Supervision,Inspection,Quarantine of the People's Republic of China in May,2006.

Objective To find out the current thyroid volume of school children and its influencing factors in Chongqing.Methods Probability sampling method(PPS) was used to select 30 counties in Chongqing in 2011.Forty children aged 8-10of 1 randomly selected school from every county chosen were enrolled in the study.Thyroid volume of children was examined by B-ultrasonograghy.Body height and body weight were measured.The relationship between gender,age,height and weight and thyroid volume was analyzed,respectively.Results One thousand three hundred and twenty-two children aged 8-10 were investigated.The median of thyroid volume was 3.53 ml.The goiter rate was 5.52％ (73/1322).Thyroid volume of female and male was 3.55 and 3.51 ml,respectively.There was no significant difference of thyroid volume between female and male (H =0.68,P ＞ 0.05).Thyroid volume of children aged 8,9 and 10 was 3.30,3.53 and 3.76 ml,respectively.There was a significant difference of thyroid volume among different age groups(H =52.49,P ＜ 0.01).Thyroid volume of children height (110-,120-,130-and ≥140 cm,respectively) was 2.96,3.22,3.59 and 4.13 ml.There was a significant difference of thyroid volume among different height groups (H =149.23,P ＜ 0.01).Thyroid volume of children weight(17-,20-,30-and ≥40 kg,respectively) was 2.71,3.31,3.91 and 4.74 ml.There was a significant difference of thyroid volume among different weight groups(H =138.44,P ＜ 0.01).For the coefficients of simple and partial correlation,there was a significant correlation between thyroid volume and age,height and weight (P ＜ 0.05).The Spearman coefficient was 0.2411,0.3950 and 0.4285,respectively.The partial correlation coefficient was 0.0640,0.1154 and 0.2319,respectively.The standard partial coefficient of age,height and weight was 0.640,0.1154 and 0.3410,respectively.The proportion of the standard partial coefficients was 1 ∶ 1.8 ∶ 5.3.The function of body weight to thyroid volume was 5.3 times that of age and 3.0 times that of body height.Conclusions The goiter rate of schoolchildren in Chongqing is relatively high.Thyroid volume is affected by age,body height and body weight.The relationship between thyroid volume and iodine nutrition needs further study.

Objective - To analyze the relationship between cobalt level of post shift urine and individual exposure level of ， cobalt and its compounds in cobalt exposed workers and to explore the feasibility of using urine cobalt as a biomarker. Methods - A total of 148 occupational cobalt exposed workers from a new material company were selected as the exposed ， - - group and 44 non occupational cobalt exposed workers from the company were selected as the control group using the typical sampling method. The exposure concentration of time weighted average of cobalt and its compounds in the workplace air of the - two groups was determined by inductively coupled plasma mass spectrometry as the individual exposure level. The cobalt levels - - of pre shift and post shift urinary samples of the two groups were detected by this method. The linear relationship between the - cobalt level of post shift urine and the individual exposure level of cobalt and its compounds in the air of the workplace was Results - 3 analyzed. The individual exposure level of cobalt and its compounds in the exposed group was 1.10 131.71 μg/m with （M） 3 the median of 12.23 μg/m. No cobalt and its compounds were detected in the workplace air in the control group. The cobalt - - levels of pre shift and post shift urines in exposed group were higher than those in the control group at the same time point （M： vs ， vs ， P ） - - 1.54 0.56 μg/L 8.77 0.83 μg/L all <0.01 . The cobalt level of post shift urine was higher than that in pre shift （M： vs ，P ）， urine in the exposed group 8.77 1.54 μg/L <0.01 and it was positively correlated with the individual exposure level （ ，P ） ， of cobalt and its compounds Spearman correlation coefficient=0.86 <0.01 . After common logarithm conversion the linear regression equation of the cobalt level of post shift urine and the common logarithm of individual exposure level of cobalt and （x） ：ŷ x（ ；F ， its compounds in the exposed group was as follows = −0.178 + 0.988 coefficient of determination=0.72 =374.75 P ；t ， P ） Conclusion - <0.01 = - 19.36 <0.01 . There was a linear correlation between cobalt level of post shift urine and occupational cobalt exposure level of cobalt exposed workers. Urinary cobalt can be used as a biomarker of occupational cobalt

OBJECTIVETo assess the relationship between fractional esterification rate of high density lipoprotein cholesterol (FER(HDL)) and coronary artery disease.

METHODSA total of 131 hospitalized patients underwent coronary angiography due to chest pain were included in the study and patients were divided into CAD group (n = 76) and non CAD group (n = 55) according to coronary angiogram. Clinical and laboratory data including biochemical laboratory, FER(HDL) and lipid subclasses were analyzed.

RESULTSThe FER(HDL) value of CAD group was significantly higher than that of the non CAD group (21.70 ± 8.73 vs. 18.65 ± 6.26, P < 0.05). There was an increased trend of FER(HDL) with numbers of diseased coronary arteries, significant difference was evidenced between non CAD group and 3-vessel group (18.65 ± 6.26 vs. 24.00 ± 9.22, P < 0.05). FER(HDL) was positively correlated with TG (r = 0.647, P < 0.001), LDLb-C(r = 0.441, P < 0.001) and negatively correlated with HDL-C (r = -0.708, P < 0.001) and HDL(2)-C (r = -0.748, P < 0.001).

CONCLUSIONOur data showed that the values of FER(HDL) were significantly increased in CAD patients and correlated with the severity of the CAD.

Adult ; Aged ; Aged, 80 and over ; Cholesterol, HDL ; metabolism ; Cholesterol, LDL ; metabolism ; Coronary Angiography ; Coronary Artery Disease ; diagnostic imaging ; metabolism ; Esterification ; Female ; Humans ; Lipoproteins, HDL ; metabolism ; Lipoproteins, LDL ; metabolism ; Male ; Middle Aged

Objective To investigate iodine deficiency disorders(IDD) in Chongqing and Linzhi, and to provide scientific basis for IDD control and prevention. Methods According to the national program developed in 2007, investigation was conducted in Chengkou and Wuxi county in Chongqing municipality, and Linzhi, Bomi,Milin and Langxian county in Linzhi prefecture. Five towns were sampled in Linzhi county, and 3 in other counties.In each town, one township primary school and two village primary schools were selected to inspect thyroid by B ultrasound and palpation, and urinary iodine of children aged 8 to 10 years was tested in these schools. Meanwhile,2 villages were selected in each town for test of salt iodine level and urinary iodine of childbearing age women and search cretin cases. Results Three hundred and forty families in Chongqing and 915 families in Linzhi were investigated. The coverage of iodized salt in Chongqing was 98.82%(336/340), which was significantly higher than that in Linzhi[66.34%(607/905), x2 = 139.56, P ＜ 0.01]. Goiter rate of children in Chongqing was 9.27%(89/960) by palpation and 8.34% (61/731) by B ultrasound, while goiter rate of children in Linzhi was 7.80%(102/1308) by palpation and 5.53% (69/1248) by B ultrasound. The difference of goiter rate by palpation between Chongqing and Linzhi was not statistically significant (x2 = 1.37, P ＞ 0.05 ). But goiter rate of children by B ultrasound in Chongqing was higher than that in Linzhi (x2= 5.51, P ＜ 0.05). In Chongqing, the median urinary iodine was 319.15 μg/L, and 345.75 μg/L in Chengkou county and 281.39 μg/L in Wuxi county. In Linzhi prefecture, the median urinary iodine was 189.81 μg/L, and 207.81 μg/L in Linzhi county, 161.12 μg/L in Bomi county, 131.83 μg/L in Milin county and 334.60 μg/L in Langxian county. The median urinary iodine in childbearing women were 248.42 μg/L in Chongqing and 121.25 μg/L in Linzhi. The median urinary iodine in Chongqing both in children and women were higher than those in Linzhi. No new cretin case was found in these two areas. Conclusions Goiter rate in high risk areas of IDD in Chongqing and Linzhi has decreased to less than 10%.No new cretin case is found in these areas. It can be concluded that the work of control and prevention is effective.There is excess iodine in Chongqing. In Linzhi county and Langxian county, iodine is excess in children and deficient in women. Further investigation should be conducted to find out the reason. Population iodine is excess in Bomi and Milin counties. The concentration of salt iodine should be decreased in Chongqing. In Linzhi prefecture,adding iodine measures should be adjusted based on further investigation.

Objective To investigate the fulfillment of improved water measures for endemic fluomsis and to find out the trend of prevalence in Qinghai Province in order to provide scientific basis and technical support for the government to formulate control strategies for endemic fluorosis.Methods Usage and management of reforming water facilities in Huzhu County were generally surveyed.Yanya Village,Caijiabu Town,Huzhu County was chosen as the surveillance spot.The household drinking water was surveyed.The dental fluorosis and urine fluoride content of children aged 8-12 years and adult above 16 years were examined.Skeletal fluorosis of adult was checked.The fluomsis content in drinking water and urine was determined with F-ion selective electrode method.The dental fluowsis was examined with Dean index.Skeletal fluorosis was diagnosed according to eountry standard(GB 16396-1996.WS 192-1999).Results The rate of water-improving was 60%(36/60)in Huzhu County.The mean of fluoride content in drinking water Was 1.25 mg/L The prevalence rate of dental fluorosis of children aged 8-12 years was 90.20%(46/51);that of adult was 88.89%(48/54).The dental fluorosis index of children was 1.77,that of adult was 2.95.The prevalence rate of skeletal fluorosis was 98.15% indicated by clinical data,18.87% by X-ray.The ufine fluorosis content of children was 2.27 mg/L,that of adult was 2.00 mg/L.Conclusion The disease condition of endemic fluorosis in Qinshai is serious,defluofidation is slow in effect.

Aged ; Animals ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Middle Aged ; Orientia tsutsugamushi ; Scrub Typhus ; epidemiology

BACKGROUNDMyocardial apoptosis is involved in the pathogenesis of sepsis-related myocardial depression. However, the underlying mechanism remains unknown. This study investigated the role of mitochondrial damage and mitochondria-induced oxidative stress during cardiac apoptosis in septic rats.

METHODSSeventy-two Sprague-Dawley rats were randomly divided into a control group and septic group receiving lipopolysaccharide injection. Heart tissue was removed and changes in cardiac morphology were observed by light microscopy and scanning electron microscopy. In situ apoptosis was examined using terminal transferase-mediated dUTP nick end-labeling assay and nuclear factor-kappa B activation in myocardium by Western blotting to estimate myocardial apoptosis. Appearance of mitochondrial cristae and activation of cytochrome C oxidase were used to evaluate mitochondrial damage. Oxidative stress was assessed by mitochondrial lipid and protein oxidation, and antioxidant defense was assessed by mitochondrial superoxide dismutase and glutathione peroxidase activity.

RESULTSSepsis-induced inflammatory cell infiltration, myocardium degeneration and dropsy were time-dependent. Expanded capillaries were observed in the hearts of infected rats 24 hours post-challenge. Compared with sham-treated rats, the percentage of cell apoptosis increased in a time-dependent manner in hearts from septic rats at 6 hours, 12 hours and 24 hours post-injection (P < 0.05). The expression of nuclear factor-kappa B p65 decreased gradually in the cytosol and increased in the nucleus during sepsis, indicating that septic challenge provoked the progressive activation of nuclear factor-kappa B. Mitochondrial cristae and activation of cytochrome C oxidase increased in a time-dependent manner. Both superoxide dismutase and glutathione peroxidase activities decreased, while mitochondrial lipid and protein oxidation increased between 6 and 24 hours after lipopolysaccharide challenge.

CONCLUSIONSSeptic challenge induced myocardial apoptosis and mitochondrial damage. Furthermore, mitochondrial damage via alteration of defenses against reactive oxygen species might play an important role in myocardial apoptosis during sepsis.

Animals ; Apoptosis ; physiology ; Male ; Mitochondria, Heart ; metabolism ; pathology ; Myocardium ; metabolism ; pathology ; Oxidative Stress ; physiology ; Rats ; Rats, Sprague-Dawley ; Sepsis ; metabolism ; physiopathology