Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.

Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.

Aliphatic amines in infant food packaging materials were extracted and concentrated by 0 . 5 mL of acidified methanol using gas purge microsyringe extraction ( GP-MSE ) . Pre-column fluorescence labeling of amines was achieved in mild conditions with 10-ethyl-acridine-2-sulfonyl chloride ( EASC ) as labeling reagent. The derivatization was carried out at 60℃ and pH 10. The derivatives were successfully separated on a Hypersil GOLD column with excitation and emission wavelengths of 262 and 430 nm, respectively. The detection limits were in the range of 0. 4-0. 6 μg/kg, and the quantitation limits were in the range of 1. 2-2. 1 μg/kg. All analytes were in good linearity in the concentration range of 2. 0-2000 μg/L with correlation coefficients of higher than 0. 998. The developed method was characterized by celerity, accuracy and high sensitivity. It was successfully applied to the determination of aliphatic amines in infant food packaging materials.

Objective To explore the relationship of the ratio of plasma adrenomedullin(AM)and endothelin-1(ET-1)with serum concentration of neuron-specific enolase(NSE)in full-term neonates with hypoxic-ischemic encephalopathy(HIE).Methods Plasma concentrations of AM,ET-1 and serum NSE from 32 full-term neonates with HIE were detected by radioimmunoassay(RIA)on the 1,3 and 7 d after parturition,30 neonates in the corresponding periods in our hospital were employed as controls.The infants with HIE were divided into mild,moderate or severe group in terms of diagnostic standard of HIE.Results 1.Plasma concentrations of AM and ET-1 in newborns with mild,moderate or severe HIE were significantly higher than that of control group at 1 d after life with a decline from 3-7 d(Pa

OBJECTIVETo assess the relationship between nodular goiter and hepatitis C virus infection.

METHODSNinety-seven cases of early treatment in patients with chronic hepatitis C were collected for analysis.Data on patient age,sex,hepatitis duration and other general information were collected.In addition, data on clinical measures of thyroid function (including T3, t4, tSH) and thyroid autoantibodies (thyroid peroxidase antibody TPO-Ab, thyroglobulin antibody Tg-Ab), as well as findings from thyroid dimensional ultrasonography were collected. One hundred and eleven cases of early treatment in patients with chronic hepatitis B and 106 eases of females 40 years old or older with high risk of nodular goiter were collected for use as controls.The relationship between nodular goiter with thyroid function, thyroid autoantibodies levels,sex,age,and hepatitis C virus infection were statistically analyzed.

RESULTSThe prevalence rates of nodular goiter in the chronic hepatitis C group, the chronic hepatitis B group and the more than or equal to 40 year-old women with high risk of nodular goiter were 53.6%,36.9% and 59.4% respectively.The prevalence rates of nodular goiter in the chronic hepatitis C group and the more than or equal to 40 year-old women with high risk of nodular goiter were significantly higber than that in the chronic hepatitis B group (x² values: 5.820 and 10.996, P < 0.05). The average age of patients with chronic hepatitis C combined with nodular goiter was significantly higher than their counterparts without goiter (F=6.408, P < 0.05),and the prevalence rate in the more than or equal to 40 year-old women with high risk of nodular goiter was significantly higher than that of their counterparts who were less than 40 years-old (60.0% vs. 23.5%; x² =7.499, P less than 0.05). The prevalence of nodular goiter in patients with chronic hepatitis C was significantly greater for females than for males (62.1% vs. 41.0%; x 2 =4.152, P < 0.05).The prevalence of nodular goiter in patients with chronic hepatitis C was also significantly higher for females more than or equal to 40 years old than for males (70.2%, 33/47 vs. 45.5%,15/33; x² = 4.952, P < 0.05).The duration of hepatitis, thyroid function and thyroid autoantibodies were similar between the patients in the chronic hepatitis C group with or without nodular goiter.

CONCLUSIONSThe patients with chronic hepatitis C had a higher prevalence of nodular goiter,with an average of up to 53.6%, than the patients with chronic hepatitis B,and the women the more than or equal to 40 years old had even higher prevalence, at 70.2%, suggesting that patients with chronic hepatitis C should be routinely examined by thyroid ultrasound. Thyroid function and thyroid autoantibodies were not correlated with prevalence of goiter among the chronic hepatitis C patients.

Objective To investigate the diagnostic criteria of thyroid solid nodules by high-frequency ultrasound,elastography and ultrasound contrast.Methods Totally 87 patients with thyroid solid nodules underwent high-frequency ultrasound,elastography and ultrasound contrast before surgery,distinguishing benign from malignant lesions was executed based on pathological results,and according to the pathological character the image features by the three measures were compared.There were 40 patients divided into a control group going through the examination by high-frequency ultrasound,and 47 ones into an observation group with the examinations by the three measures.Pathological results were used as gold standard,and the two groups were compared on diagnostic sensitivity.Results High-frequency ultrasound showed significant differences of benign and malignant nodules in equal echo,very low echo,irregular peripheral halo,rates of clear nodule boundary and aspect ratio higher than as well as microcalcification (x2--4.603,17.394,5.833,14.832,10.107 and 27.392,and all Pvalues were less than 0.05).There were 85％ benign nodules gained grade Ⅰ and Ⅱ elastography images,81.82％malignant nodules had grade Ⅲ and ⅣV elastography images,and there were significant differences between benign and malignant nodules (x2=53.42,P=0.00).Ultrasound contrast found annular enhancement in most of benign nodules (x2=39.16,P＜0.01) while uneven enhancement in most of malignant nodules (x2=40.28,P＜0.01),and the nodule ranges increasedsignificantly after contrast (t=2.19,P＜0.05).The observation group had the sensitivity being 82.86％,which was statisticallyhigher than 59.38％ in the control group (x2=4.537,P--0.033).Conclusion High-frequency ultrasound,elastography and ultrasound contrast all show the manifestations of thyroid solid nodules,and the comprehensive application of the above three measures contributes to determining the nature of the nodules.

Objective To evaluate the effect of methylene blue (MB) on hydrogen peroxide (H2O2)-induced apoptosis in macrophages through mitochondria-dependent pathway in mice.Methods Mouse peritoneal macrophage line RAW264.7 cells were cultured in DMEM culture medium containing 10％ fetal bovine serum.Cells were divided into 6 groups (n =24 each) using a random number table method:control group (group C),H2O2 group,prophylactic different concentrations of MB groups (MB1,2 groups) and therapeutic different concentrations of MB groups (MB3.4 groups).H2O2 50 μmol/L was added to the culture medium in group H2O2.MB was added to the culture medium with the final concentrations of 0.1 μmol/L (in MB1 and MB3 groups) and 1.0 μmol/L (in MB2 and MB4 groups) at 30 min before adding H2O2 in MB1.2 groups and 30 min after adding H2O2 in MB3.4 groups.At 24 h of culture or incubation in each group,the cell survival rate was measured by methyl thiazolyl tetrazolium assay,the activity of reactive oxygen species (ROS) in cells was determined with the fluorescent probe,the lactate dehydrogenase (LDH) activity in supernatant was detected by spectrophotometry,the activity of superoxide dismutase (SOD) in cells was detected by colorimetric method,mitochondrial membrane potential (MMP) was measured using rhodamine 123 staining,the content of ATP was determined by an ATP bioluminescent method,the expression of pro-caspase-3 and spliceosomes P20 protein and P 18 protein was detected by Western blot,and cell apoptosis was detected using flow cytometry.Results Compared with group C,the cell survival rate,SOD activity and contents of MMP and ATP were significantly decreased,the ROS activity and activity of LDH in supernatant were increased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was up-regulated,and early and late apoptosis rates were increased in the other five groups (P＜0.05).Compared with group H2O2,the cell survival rate,SOD activity and contents of MMP and ATP were significantly increased,the ROS activity and activity of LDH in supernatant were decreased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was down-regulated,and early and late apoptosis rates were decreased in MB1-4 groups (P＜0.05).Compared with group MB1,the cell survival rate was significantly decreased,and the expression of caspase-3 spliceosome P 18 was down-regulated in group MB2,and the cell survival rate and SOD activity were significantly decreased,and the activity of ROS was increased in group MB3 (P＜0.05).Compared with group MB4,the expression of caspase-3 spliceosome P 18 was significantly down-regulated,early and late apoptosis rates were decreased,and the activity of ROS was increased in group MB2,and the activity of ROS was significantly increased in group MB3 (P＜0.05).Conclusion The mechanism by which MB attenuates H2O2-induced oxidative damage to macrophages is related to inhibiting cell apoptosis in macrophages through mitochondria-dependent pathway in mice.

Bone Wires ; Child ; Child, Preschool ; External Fixators ; Female ; Fracture Fixation ; methods ; Humans ; Humeral Fractures ; surgery ; Male

A novel qualitative analytical method by using two-dimensional chromatographic correlation spectroscopy techniques for recognizing impurity peaks of HPLC methods of quality control and LC-MS chromatographic system was established. The structures of major degradation products of ceftizoxime and cefdinir were identified by LC-MS and MassWorks application; the standard chromatographic and spectral data of the degradation impurities were obtained by high-performance liquid chromatography with diode array detection. The impurity peaks of two-dimensional chromatography were matched by comparison of spectra and calculating correlation coefficients. Peaks in chromatography can be identified accurately and rapidly in different chromatographic systems such as column and mobile phase changed. The method provides a new way and thought to identify the peaks in quality control of impurities without reference impurity substances.
Ceftizoxime ; analysis ; chemistry ; Cephalosporins ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Liquid ; methods ; Drug Contamination ; Mass Spectrometry ; methods ; Quality Control

Objective: To analyze the clinical characteristics of hepatic flare and evaluate efficacy of antiviral treatment in pregnant women with chronic HBV infection. Methods: A single-center, open-label, prospective study was conducted, and pregnant women with chronic HBV infection were enrolled. Liver function, HBV serum markers and HBV DNA of pregnant women with chronic HBV infection were reviewed during every 4 to 12 weeks of gestation period. The proportion and clinical characteristics of hepatitis flare during pregnancy were observed. Logistic regression analysis was used to predict hepatic flare in pregnant women with chronic HBV infection. Antiviral therapy with telbivudine (LdT) or tenofovir dipivoxil (TDF) was used to treat hepatic flare during pregnancy. Sequential entecavir (ETV) or TDF was applied after the delivery. Treatment course and drug withdrawal in pregnant women with hepatic flare was the same as those of the general patients with chronic hepatitis B. Liver function, HBV serum markers and HBV DNA were measured in pregnant women with hepatic flare at different time points (4, 12, 24 and 52 weeks). A t-test was used to compare the hepatic flare in pregnant women with and without hepatitis group. HBsAg and HBeAg were used to quantify the receiver operating characteristic (ROC) curve of pregnant women with hepatic flare during pregnancy. Area under the ROC curve was used to calculate the optimal cut-off value corresponding to the maximum sensitivity and specificity of the ROC curve. Results: Of the 220 pregnant women with chronic HBV infection, 55 (25%) had hepatitis flare during pregnancy and received antiviral treatment. Among the 55 women with hepatic flare during gestation, 47 (85.46%) had hepatic flare in the mid-second trimester (12-24 weeks); average peak value of alanine aminotransferase (ALT) was 220.62 U/L, and the average peak value of ALT in 32 cases (58.18%) of pregnant women with hepatic flare was between 2–5 × ULN. HBsAg and HBeAg quantification were significantly lower in pregnant women with hepatic flare during pregnancy than with non-hepatitis (t = -3.745, P < 0.001; t = -2.186, P = 0.030). Multivariate logistic regression analysis showed that pregnant women with HBeAg < 3.065 log10 s/co were 7.576 times more likely to have hepatic flare during pregnancy (95% confidence interval: 3.779-15.190). ALT normalization, undetectable HBV DNA levels, HBeAg loss and HBeAg seroconversion in 55 pregnant women with hepatic flare at 52-week treatment was 100% (55/55), 74.55% (41/55), 47.27% (26/55) and 41.82% (23/55), respectively. HBsAg quantification at 52 weeks was significantly lower than baseline HBsAg quantification (3.32 + 0.37) log₁₀ IU/ml and (3.95 + 0.40) log₁₀ IU/ml; t = 8.465, P < 0.001). Conclusion Hepatic flare often occurs in the second trimester of pregnancy in pregnant women with chronic HBV infection and baseline HBeAg quantification is an independent predictor of hepatic flare. HBeAg seroconversion rate increased at 52 weeks after antiviral therapy.