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.

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

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

Objective To investigate expression and significance of decorin(DCN)in liver tissue and serum of liver transplant patients with chronic rejection(CR).Methods Immunohistochemistry(SP method)was used to detect expression of DCN in liver tissue of 16 normal controls, 20 patients with cirrhosis, 46 liver translantion patients without CR and 8 patients with CR.Enzyme-linked immunosorbent assay method(ELISA method)was used to determined the content of DCN in serum of all research subjects.Results The expression of DCN was negative in normal hepatic tissues and with/without CR, cirrhosis tissues showed strong expression of DCN.The positive expression rate and the average optical density value of DCN in liver transplant tissues with CR had significant difference comparing with Cirrhosis tissues(25% vs 55%, 0.1249 ±0.0039 vs 0.2357 ±0.0396, P <0.01,while no statistic siqnificance compared to normal liver tissues and those without CR.The level of DCN in serum was significantly higher in liver transplant patients with CR, with significant difference comparing with normal people, liver cirrhosis and transplant liver patients without CR(54.0833 ± 6.0325)μg/L vs(1.0232 ± 0.9105)μg/L,(12.6202 ± 1.5370)μg/L,(17.7102 ± 2.3562)μg/L, P < 0.01).The concentration of DCN in serum showed a positive correlation with the degree of CR.Conclusions DCN showed negative expression in liver tissue and increased significantly in serum of liver transplantation patients with CR.This suggests that DCN may be involved in occurrence and development of CR.At the same time the determination of DCN in serum maybe become an important indicator of the early diagnosis, development and prognosis of CR for liver transplant patients.

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.

Objective To evaluate the feasibility and safety in the treatment of liver cancer located under the diaphragm with cool-tip radiofrequency ablation(RFA)percutaneously under CT guidance.Methods 20 patients with total 25 lesions were treated by CT-guided RFA with cool-tip electrode involving the induced necroses.The postoperative efficacy was evaluated by enhanced CT or MRI.Results 72% lesions were completely necrotized(18/25),28% lesions were majorly necrotized(7/25).No severe complications occurred. Conclusion CT-RFA with cool-tip electrode is effective and safe in treating liver cancer located under the diaphragm.