Objective To construct the vector pET15b-Z-VP1 by inserting the Z fragment into amino-terminal of JCV VP1.Methods The VP1 and Z fragment were amplified by PCR from plasmid pET15b and pEZZ18 respectively,and then they were linked by recombinant PCR.The Z-VP1 fragment was inserted into plasmid pET15b by restriction enzyme BamHⅠ and NcoⅠ.Results The VP1 and Z fragment were obtained by PCR and gel purification.The Z-VP1 fragment,which was linked by recombinant PCR from VP1 and Z fragment,was inserted into plasmid pET15b between BamHⅠ and NcoⅠ sites,and confirmed by enzyme digestion and DNA sequencing.The expression of VP1-Z was confirmed by Western blotting.Conclusion The plasmid pET15b-Z-VP1 has been constructed successfully by inserting Z fragment into amino-terminal of VP1.

Objective To express the recombinant protein VP1-Z, and investigate whether VLP-Z has the physiological functions like as wild-type VLP. Methods The expression plasmid pET15b-VP1-Z was introduced into competent E. coil BL21 (DF3)/pLys cells by transformation, and the expression of re-combinant protein VP1-Z was induced by incubation of the cells with IPTG. The protein was prepared as pre-viously described for wild-type VLP. The morphous of VLP-Z were observed by electron microscopy, and the physiological functions of VLP-Z were investigated by hemagglutination test and by immunofluorescence. Re-sults The purified VLP-Z composed of VP1-Z possessed hemagglutination activity and yielded a prominent band of 50×10~3 on SDS-PAGE and staining with Coomassie Brilliant Blue. The VLP-Z exhibited virus-like particles structure like as wild-type VLP with a diameter of 45-50 nm, which was slightly bigger than that of wild-type VLP(42-45 nm). In immunofluorescence test, VP1-Z was detected within the cytoplasm and nu-cleus after HeLa cells were inoculated with VLP-Z. Conclusion The physiological functions of recombinat-ed protein VLP-Z were comparable with wild-type VLP.

Objective To investigate the effects of lead exposure on rat placental apoptosis during different periods of pregnancy. Methods All Wistar rats were randomly divided into four groups with 27 for each group with the sex ratio of 2 : 1 (female : male).The groups with lead exposure consumed water with 0. 025％ lead acetate during the entire, early or late period of pregnancy. Controls were given distilled water without lead.Blood lead levels were determined by atomic absorption spectrophotometry at the end of pregnancy. Placental apoptosis were assessed by both Hoechst staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay. Data were assessed by one-way analysis of variance, and differences between groups were compared by q-test. ResultsBlood lead levels at the end of pregnancy of the groups with lead exposure (entire, early or late pregnancy) and the control group were (1. 74±0. 19) μmol/L,(1.27±0.26) μmol/L, (0.60±0. 11) μmol/L and (0.04±0.01) μmol/L respectively(F= 12. 10,P＜0.01).In the groups with lead exposure, Hoechst staining showed hyperchromatic nuclei in placental trophoblast apoptotic cells and compact fluorescent particles in some nucleus； TUNEL assay showed brown-staining apoptotic cells nuclei with some nuclei particles staining brown. Two assays showed the same results: the apoptotic index of the groups with lead exposure were higher than that of the control group; the apoptosis index of the group with lead exposure during entire pregnancy was higher than that of the group with lead exposure during early and late pregnancy (P ＜ 0. 05).ConclusionsLead exposure during pregnancy could elevate the blood levels of lead and the degree of placental apoptosis.

Objective To observe the changes in the transforming growth factor-beta 1 (TGF-β1 ) mRNA expression in the lung in a dog model of cardiopulmonary bypass(CPB)-induced acute lung injury. Methods Thirty-six healthy adult mongrel dogs of both sexes weighing 15-16 kg were randomly assigned into control group and CPB group ( n = 18 each) . Lung injury was produced by CPB according to the method described by Williams. Six animals were killed at each of the following time points: before CPB (T0 ) and 30 and 60 min after termination of CPB (T1 , T2) in each group. Lung specimens were obtained for microscopic examination and determination of TGF-β1 mRNA expression (by RT-PCR) and MDA content. The lungs were lavaged and the protein concentration in the brancho-alveolar lavage fluid (BALF) was determined and pulmonary permeability index (PPI) was calculated. Results Microscopic examination showed massive inflammatory cell infiltration, alveolar capillary dilatation, congestion, widened alveolar septum, massive RBC in the alveolar space and focal atelectasis in the lung in CPB group. The TGF-β1 mRNA expression and MDA content and PPI were significantly higher in CPB group than in control group. The TGF-β1 mRNA expression and MDA was positively correlated to PPI (MDA: r = 0.867, P < 0.01; PPI: r = 0.821, P < 0.01) . Conclusion TGF-β1 mRNA expression in the lung is significantly up-regulated after CPB and is an important factor contributing to CPB-induced acute lung injury.

Objective To investigate the effect of enteral nutrition support on prognosis in patients with severe acute stroke.Methods Ninety-eight cases of severe acute stroke were divided into nutrition supported group(50 cases) and control group(48 cases) randomly.All patients were treated by conventional therapy.Nutrition supported group received high energy nutrition diet and control group received common liquid diet by nasal feeding.The levels of hemoglobin,serum albumin,neurologic impairment score (NIHSS)and the incidence rates of complications were assessed at the 1st,10 th and 21st day after admission.Results Hemoglobin was (122.5 ± 2.4),(106.4 ± 2.8) g/L and serum albumin was (36.5 ± 4.7),(34.2 ± 5.1) g/L at the 21st day after admission in nutrition supported group and control group,and there was significant difference between two groups (P < 0.05).The complication rate in control group was higher than that in nutrition supported group (P< 0.05).NIHSS score was(8.45 ± 3.02) scores in nutrition supported group and (10.24 ± 2.57)scores in control group at the 21st day after admission,and there was significant difference between two groups (P <0.05).Conclusion Early enteral nutrition support can significantly improve nutritional status and decrease the incidence rates of complications in patients with severe acute stroke,and it is helpful for clinical prognosis.

Objective To investigate the prevalence of malnutrition and nutritional support and interventions in geriatric inpatients.Methods The elderly patients (aged ≥ 65 years)from the geriatric demonstration ward were consecutively enrolled from July 2010 to January 2012.MiniNutritional Assessment-short form (MNA-SF) was performed after admission,and data of nutritional support were collected.Results A total of 179 patients were enrolled in this study.According to MNA-SF,42 cases (23.5％)were rated as malnutrition,and 55 cases (30.7％) were rated as at risk of malnutrition.Totally,45 patients received nutritional support.50.0％ (21/42) patients with malnutrition,and 29.1％ (16/55) patients at risk of malnutrition received nutritional support.As to the route of nutrition therapy,the ratio of the enteral to parenteral to combination of enteral and parental nutrition was 4.4 ∶ 1.0 ∶ 1.0.Conclusions The incidence of malnutrition is high in the geriatric inpatients,and routine nutritional risk screening and assessment are essential for the elderly patients.Nutritional support and other comprehensive treatment are in great need,and the enteral nutrition is appropriate and preferred.

ObjectiveTo study the different clinical effects of using 5 kinds of hemostatic surgeries to manage the intractable postpartum hemorrhage and analyse the risk factors of failed hemostasis.Methods From Jan.2007 to Jul.2011,96 patients with intractable postpartum hemorrhage were studied retrospectively and grouped by the first step surgical treatment.The hemostatic surgeries included uterine tamponade (tamponadegroup ), pelvicbloodvessels ligation(ligationgroup), pelvical arterial embolization (embolization group), uterine compression sutures (sutures group)and uterine compression sutures combining tamponade (combined group).The intraoperative and postoperation datum were compared among groups,so dose the treatment outcomes.Multivariate analysis were used for failed hemostasis.Results( 1 ) The blood loss of 96 patients ranged from 1200 to 9100 ml,and 71 patients had a succeed hemoatasis after employing these surgeries and 25 failed.(2) The blood loss before hemostasis surgeries in tamponade group and embolization group was statisically greater than in sutures group ( P ＜ 0.05 ).Blood loss during the hemostasis surgeries in ligation group was statistically greater than in embolization and sutures groups ( P ＜0.05).The operating time of embolization group was statistically shorter than ligation group,sutures group and the combined group (P ＜ 0.05 ).(3) Fine of 96 patients had uterine atony and 43 had a successful hemostasis with the success rate about 78％.Forty-six had placenta previa and 39 success with success rate 85％.Thirty-three had placenta accrete and 13 of which succeed in hemostasis with success rate about 39％.In patients with uterine atony and placenta previa,the difference of hemostasis rate in groups had no statistically significant ( P ＞ 0.05 ).In patients with placenta accrete,the hemostasis rate in embolization group was higher than in others groups (P ＜ 0.01 ). (4) The multivariate analysis found that scar uterus,placenta accrete and coagulation defects were the risk factors of failed hemotasis.The OR value respectively was 2.9 (95 ％ CI:1.1 - 7.6 ),17.9 ( 95 ％ CI:5.6 - 56.3 ) and 16.2 ( 95 ％ CI:3.2 - 83.5 ).Embolization had some extent of protective effection ( OR =0.9,95 ％ CI:0.8 - 0.9 ).Conclusions ( 1 ) Five kinds of hemostatic surgeries were all effective.Though the success rate among groups did show statistical difference,pelvical arterial embolization has the comparative advantage of shorter operating time,less operating blood loss and higher success rate in placenta accrete.(2) Since scar uterus,placenta accrete and coagulation defects were the risk factors of failed hemostasis,sufficient preparation should be made for patients with these risk factors and the hemostatic surgeries should be choosed individually.

Objective To study the inhibitory effect of arsenic trioxide(As_2O_3) on the tumor growth of breast cancer cell line MCF-7 implanted subcutaneously in nude mice and its mechanism.Methods BALB/C-nu/nu nude mice were subcutaneously injected with MCF-7 breast cancer cell line,and treated with intraperitoneal injection of As_2O_3 and 5-FU in different concentrations.The implanted tumor was weighed,and the tumor inhibition rates were calculated.The apoptosis of the implanted tumor was detected by flow cytometry.The expressions of bcl-2 and Fas induced by As_2O_3 were examined by immunohistochemical method.Routine blood test and bone marrow test were used to observe the function of hematopoietic system after As_2O_3 treatment.Results The growth of implanted tumor was markedly inhibited with 5-FU,low dose and high dose As_2O_3,the inhibitory rates being 38.33%、51.42% and 62.43%,respectively.The inhibitory effect of As_2O_3 was significantly stronger than that of 5-FU(P

Objective To observe the phenomenon of apoptosis and expression of related proteins in injured muscle tissue during the formation of pressure injury, and to explore its mechanism of action on the pressure injury. Methods Forty male Sprague-Dawley (SD) rats were divided into normal control group, 3, 5, 7 and 9 compression groups according to the random number table, with 8 rats in each group. The pressure injury models on the gracilis muscle of hind limbs were reproduced by using a way of cycle compression of ischemia/reperfusion (I/R) magnet. One cycle consisted of 12-hour compression, and followed by 12-hour release. The 5, 7, 9 compression groups were cut through after receiving 3 cycles. The normal control group did not receive any treatment. Muscle tissue specimens were harvested in the pressurized center at the end of the experiment (the same site for the normal control group), then the rats were sacrificed. The hematoxylin-eosin (HE) staining was used to examine the changes of muscle tissue morphology. The Hoechst 33258 staining was used to evaluate the apoptosis of muscle tissue. Western Blot was used to detect the expressions of heat shock protein 70 (HSP70), B-cell lymphoma-2 protein (Bcl-2) and Bcl-2 associated X protein (Bax). Results ① HE staining showed that the compressed tissues appeared different degrees of pathological degradation, and the change of tissue morphology was more serious with the increase of the compression cycle.② Hoechst 33258 staining showed that the nuclei of compressed tissue showed condensed, compact morphology and granular fluorescence, and the number of apoptotic cells increased with the increase of the compression cycle.③ Western Blot showed that with the increase of the compression cycle the protein expressions of HSP70 and Bax were gradually increased, and the protein expression of Bcl-2 was gradually decreased. Compared with the normal control group, the protein expression of HSP70 in the 9 compression group was increased with statistically significant differences (HSP70/GAPDH: 1.78±0.21 vs. 0.55±0.17, P < 0.01). The protein expression of Bax in the 7 and 9 compression groups were increased with statistically significant differences (Bax/GAPDH: 0.96±0.09, 0.98±0.02 vs. 0.67±0.07, both P < 0.01). The protein expression of Bcl-2 in the 3, 5, 7 and 9 compression groups were significantly decreased (Bcl-2/GAPDH: 0.17±0.03, 0.13±0.03, 0.14±0.03, 0.10±0.02 vs. 0.36±0.04, all P < 0.05). Conclusions Apoptosis can be induced by I/R in pressure injury tissues. Apoptosis induced by HSP70 and apoptosis factors Bcl-2 and Bax may be involved in the formation of pressure injury.

Objective This study aimed to survey students' satisfaction and cognitive attitude of elaborate course of social medicine to provide a basis for exploring appropriate teaching mode. Methods Purposive sampling was used with questionnaire to survey 646 students who had learned social medicine in 5 universities in Guangdong province. The survey included the social medicine curriculum, teaching situation, students' satisfaction and cognitive attitude of elaborate course of so-cial medicine. Descriptive statistical analysis was used to describe the demographic characteristics and curriculum. The t test and one-way ANOVA were used to compare the means of students' satisfaction and cognitive attitude of elaborate course in different universities, using the test level of 0.05. Results The lecture-based learning was the main teaching mode. 335(52.7%) students thought it necessary to learn social medicine, but only 27.2%of the students(173 students) were interested in social medicine and 65(10.2%) students could take the initiative to read about extracurricular books. Overall satisfac-tion with using network resources of elaborate course in two universities that had university-level elab-orate course of social medicine were (2.78 ± 0.55) and (3.01 ± 0.30) respectively and the difference was statistically significant(t=-3.97,P<0.01). The cognitive attitude of the students in 5 universities with using network resources of elaborate course was (3.09±0.66) and the difference was statistically significant(F=3.70,P<0.01). Conclusion Social medicine teaching has been paid attention to in 5 universities, but there were some common problems: ①putting emphasis on theory and ignoring prac-tice in teaching mode; ②putting emphasis on credit and ignoring effectiveness in learning; ③and putting emphasis on construction and ignoring using in network resources of elaborate course of social medicine.