Objective To study the correlation between the blood selenium level and glutathione peroxidase(GSH-Px)with the occurrence of gestational diabetes mellitus(GDM).Methods Among 1 360 pregnant women in our hospital from January 2011 to December 2012,68 cases GDM were diagnosed as GDM (observation group)and other 113 non-GDM pregnant women were ran-domly selected from these 1360 pregnant women as the control group.The serum selenium level and GSH-Px activity were com-pared between the two groups.Results The selenium level and GSH-Px activity in the control group were higher than those in the observation group.The selenium level was (3.07±0.51)mmol/L in the control group and (2.06±0.41)mmol/L in the observa-tion group,respectively;the GSH-Px activity was (197.23±18.73)U in the control group and (151.35±19.67)U in the observa-tion group,the differences between the two groups were statistically significant (P <0.05).Conclusion The decrease of serum se-lenium level and GSH-Px activity may be one of the pathogenic causes of GDM.

OBJECTIVE:To observe the efficacy and safety of carboprost and Foley catheter for the expand cervix before hys-teroscopic electric resection. METHODS:80 patients with hysteroscopic operation were randomly divided into research group and control group. Research group was placed inside the vagina carboprost 1 mg 30 minutes before operation;control group was placed inside the Foley catheter 12 hour before operation. The relaxation degree of the cervix,average operative time,preoperative pain, postoperative morbidity and incidence of adverse reactions in 2 groups were observed. RESULTS:There were no significant differ-ences in the relaxation degree of the cervix,postoperative morbidity and incidence of adverse reactions between 2 groups(P>0.05). The average operative time and preoperative pain in research group were significantly better than control group,the differ-ence was statistically significant(P<0.05). CONCLUSIONS:Carboprost and Foley catheter have good efficacy and safety for ex-pand cervix before hysteroscopic electric resection,however,carboprost is better than Foley catheter in terms of average operative time and preoperative pain.

Objective: To express the protein of HPV18E6 based on pET-32a(+) at high level and study the expression and significance of HPV18E6 proteins in laryngeal carcinoma. Methods: The HPV18E6 gene was amplified by PCR and cloned into pET-32a(+). The amplified fragment was inserted into the plasmid pET32a (+) that was digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET32/E6 was transformed into E.coli JM109 which was selected with ampicillin. The recombinant plasmids were successfully introduced into E.coli BL21(DE 3) and were induced by IPTG. SDS-PAGE and Western blot analysis were used to detect the confusion protein. Finally, the optimization of expression conditions, such as temperature, concentration of IPTG, was studied. Results: The recombinant plasmids were identified and confirmed with enzyme digestion and sequencing. The BL21(DE3) transformed recombinant plasmid pET32/E6 had expressed HPV18E6 recombinant protein effectively. The optimum conditions of expression were 37 ℃, 1 mmol/L IPTG. Conclusion:Prokaryotic expression vector pET-32a(+)-HPV18E6 was successfully constructed. The high-level expression of HPV18E6 was achieved in E.coli BL21(DE3).

Objective To evaluate the clinical and radiological results of locking plate for treatment of proximal humerus frac‐tures and the efficacy of anatomical healing of tuberosities .Methods A total of 57 patients with proximal humerus fractures were treated with locking‐plate from July 2008 to March 2012 .A standardized radiological evaluation was conducted .Patients were divid‐ed into two groups :group A (anatomical healing of tuberosities) with 31 cases and group B (without anatomical healing of tuberosi‐ties) with 26 cases .Clinical assessment was performed using the Neer rating scale .Results Considering the entire sample ,the mean Neer score was 87 .96 ± 5 .06 points ,the excellent rate was 94 .74℅ .Comparing these parameters in the two groups ,group A was significant higher in the Neer scores and the range of movement than that of group B (P<0 .05) ,there was no statistic differences in the scores of pain and function (P>0 .05) .Conclusion the locing plate for treatment of complex proximal humerus fractures has a high subjective satisfaction rate .A good functional result depends on anatomical reestablishment of proximal humerus anatomy , particularly the healing of the greater tuberosity .

Prokaryotic expression vector of mouse HPV16E6 gene was constructed. A pair of primers were designed according to the digestion sites in plasmid pGEX-KG and the HPV16E6 gene sequence published by GenBank. The DNA fragment of 321bp was amplified by PCR from the HPV recombinant plasmid with HPV16E6 gene, then cloned into pGEX-KG and transformed into the host E. coli strain JM109. The fragment was conformed to the original sequence, which indicated that fusion expression vector pGEX-KG-HPV16E6 was constructed. The pGEX-KG-HPV16E6 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degrees C, the expression product of HPV16E6 gene was identified by SDS-PAGE and Western blot. HPV16E6 fusion protein had been expressed successfully in the form of inclusion bodies, the molecular weight of fusion protein being 38 kD. Meanwhile, the optimum condition of HPV16E6 fusion protein expression was induced with 1.0 mmol/L IPTG for 4h. The fusion protein reacted specifically with the antibodies against HPV16E6. HPV16E6 gene was successfully expressed in E. coli, which could be used as a basis for preparing HPV16E6 vaccine in human.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Repressor Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To evaluate the clinical effect of milrinone nebulized inhalation for improving intraoperative cardiac function and pulmonary arterial pressure in the patients with chronic obstructive pulmonary disease (COPD) complicating pulmonary hypertension (PH).Methods Forty-four surgical patients with COPD complicating PH in the Chongqing Municipal Medical Emergency Center from June 2015 to June 2016 were chosen,including 23 cases of thoracic surgery,13 cases of abdominal surgery and 8 cases of lower extremity fracture surgery.The patients were divided into the control group and treatment group,22 cases in each group.The control group received the routine comprehensive treatment.In addition receiving the conventional comprehensive treatment,milrinone nebulized inhalation in the treatment group was given before general anesthesia.Both of the two groups were imbedded with floating catheter in the right internal jugular vein for detecting the clinical indexes:cardiac output (CO),pulmonary artery systolic pressure (PASP),pulmonary artery mean pressure (PAMP) and pulmonary capillary wedge pressure (PCWP).Results Compared with before treatment,CO after treatment in the treatment group was significantly increased (P＜0.05),PASP,PAMP and PCWP after treatment in the treatment group were significantly decreased(P＜0.05).CO,PASP,PAMP and PCWP in the control group had no statistical difference between before and after treatment,the difference was not statistically significant (P＞0.05).Conclusion Intraoperative milrinone nebulized inhalation in the patients with COPD complicating PH can effectively improve the patient's cardiopulmonary function.

ObjectiveTo investigate the effect and underlying molecular mechanism of astragaloside-Ⅳ （AS-Ⅳ） on autophagy and apoptosis of nasopharyngeal carcinoma cells. MethodIn experiments in vitro， the effect of AS-Ⅳ on the autophagy of nasopharyngeal carcinoma cells was observed by monodansylcadaverine （MDC） staining and transmission electron microscopy （TEM）. In experiments in vivo， immunofluorescence （IF） and Western blot were used to detect the changes in autophagy and apoptosis and the expression of key proteins in the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway after the establishment of a xenograft tumor model in nude mice. ResultAfter 5-8F cells were treated with AS-Ⅳ of different doses （5， 10， 20 μmol·L^-1）， the fluorescence intensity of autophagy in AS-Ⅳ groups significantly increased as compared with that in the blank group. The fluorescence expression of autophagy in AS-Ⅳ groups was the strongest after intervention for 24 hours， and the fluorescence expression in the 10 μmol·L^-1 AS-Ⅳ group was the most obvious. The autophagy activator rapamycin （RAPA） induced more autophagosomes in 5-8F cells under the transmission electron microscope， and 3-methyladenine （3-MA）， an autophagy inhibitor， did not induce autophagosome formation in 5-8F cells under the transmission electron microscope as compared with the results in the blank group. In the 10 μmol·L^-1 AS-Ⅳ group， the intracellular structure and cell membrane were intact and clear， and autophagosome formation was observed. Compared with the blank group， the AS-Ⅳ groups showed inhibited tumor volume （P<0.05， P<0.01）， potentiated fluorescence signals of microtubule-associated protein l light chain 3 type Ⅱ/microtubule-associated protein l light chain 3 type Ⅰ （LC3 Ⅱ/Ⅰ） and cleaved Caspase-3 （P<0.05， P<0.01）， increased expression levels of the mammalian homolog of yeast ATG6 （Beclin-1）， LC3 Ⅱ/Ⅰ， cleaved Caspase-3， and cleaved PARP （P<0.05， P<0.01）， down-regulated expression of ubiquitin-binding protein （p62） （P<0.05， P<0.01）， and reduced protein expression levels of phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， and phosphorylated mTOR （p-mTOR） （P<0.05， P<0.01）. ConclusionAS-Ⅳ can induce autophagy and apoptosis of nasopharyngeal carcinoma cells， and the mechanism is presumably attributed to the activation of the PI3K/Akt/mTOR signaling pathway.