AIM: To develop a simple, convenience, and inexpensive method on primary culture model of pancreatic islets in rats for the study of anti-diabetic drugs. METHODS: The pancreases of SD rats were separated from the pancreatic duct with cold Hank' s solution and picked. Then the pancreases were cut into pieces and repeatedly digested by collagenase at 37℃ for the short durations of the experiment. The isolated islets were identified by dithizone staining and the viability was evaluated by trypan blue staining. Pancreatic islets were incubated in RPMI 1640 or DMEM for 14 - 16 h, then they were transferred to new culture plates with the same medium mentioned above. Determination of insulin content of su-pernate and cell lysate and the experiment of insulin secretion by stimulation of glucose and implantation of micewith STZ-induced diabetes were used for evaluated the function of islets. RESULTS: The viability of isolated pancreatic islets was more than 95% and the purity of cultured islets was about 85% . The insulin synthesis, secretion and sensitivity of islets stimulate by glucose which were cultured in RPMI 1640 were higher than that in DMEM. The levels of blood glucose recovered to normal in type 1 diabetic mice after islets implantation. CONCLUSION : The islets got in this study have higher purity and viability with the normal biological activity for about 7 days by this method and they can be used as a cell model for the study of diabetes in vitro .

Objective To evaluate the capabilities of disc diffusion and Vitek2-compact GN13 methods for testing antimicrobial susceptibility of screening ESBLs ( extended-spectrumβ-lactamase) in En-terobacteriaceae clinical isolates.Methods A total of 93 Enterobacteriaceae strains were isolated from pa-tients with intra-abdominal infections in 21 hospitals during 2011 to 2012.The in vitro minimum inhibition concentration ( MIC ) values of ampicillin-sulbactam, piperacillin-tazobactam, ertapenem, ceftazidime, ceftriaxone, cefepime, imipenem, amikacin, ciprofloxacin and levofloxacin were determined by disc diffu-sion, Vitek2-compact GN13 and broth microdilution methods, respectively.Categorical agreement ( CA ) rates of disc diffusion and Vitek2-compact GN13 methods were determined by using broth microdilution meth-od as the reference method.The genes encoding ESBLs were screened in Escherichia coli (E.coli), Kleb-siella pneumoniae (K.pneumonia), Klebsiella oxytoca (K.oxytoca) and Proteus mirabilis (P.mirabilis) strains by using PCR analysis and gene sequencing.Disc diffusion and Vitek2-compact GN13 methods were used for the phenotypic confirmatory test of ESBLs and the sensitivity, specificity, positive predictive value and negative predictive value of the two tests were evaluated.Results The CA values of disc diffusion and Vitek2-compact GN13 methods for the 10 antibiotics were all >90% as compared with broth microdilution method.The major error (ME) rate for ertapenem was 3.2%and the very major error (VME) rates for am- picillin-sulbactam, ceftazidime and cefepime tests were all 2.2% by using Vitek2-compact GN13 method. The sensitivity, specificity, positive predictive value and negative predictive value of disc diffusion and Vitek2-compact GN13 methods in the phenotypic confirmatory test of ESBLs were 96.7%(29/30), 100%(20/20), 100%(30/30) and 95%(19/20), respectively.Conclusion Both disc diffusion and Vitek2-compact GN13 methods could be used for testing the antimicrobial susceptibility and the detection of ESBLs in Enterobacteriaceae clinical isolates with the advantage of accuarcy.Attention should be paid to the posibil-lity of oaurance of ME and VME when testing ertapenem, ampicillin-sulbactam, ceftazidime and cefepime by using Vitek2-compact GN13 method.

Objective To analyze the dynamic changes of perinatal defects in Lishui city from 2005 to 2014.Methods According to hospital-based data,87 294 fetuses and infants aged from 28 weeks of gestation to 7 days after birth were monitored for birth defects in Lishui city from 2005 to 2014.Results Among 87 294 perinatal infants,birth defects were found in 1 914 cases with a incidence of birth defects of 219.3/ten thousand,which was on the rise year by year except 2009.The incidence of birth defects was 235.1/ten thousand (737/31 352) in urban areas,which was significantly higher than that in rural areas (210.4/ten thousand,1 177/55 942) (x2 =5.705,P ＜ 0.05).The incidence of birth defects of boys and girls was 221.1/ten thousand (1 005/45 453) and 212.8/ten thousand (890/41 822),respectively (x2 =0.707,P ＞ 0.05).Among 1 914 cases of defects,657 were detected prenatally and 1 257 case were detected within 7 days after birth.Congenital heart disease was the most common birth defect,followed by polydactyly,cleft lip,microtia and syndactyly.Birth defect accounted for 32.74％ (278/849) of all perinatal deaths and the ratio was on the rise since 2010.Conclusion There is an upward trend in incidence of birth defects in Lishui city in the last decade;the incidence of birth defects in urban areas is significantly higher than that in rural areas;the incidence of congenital heart disease is high,and birth defects are the main cause of perinatal death.

Objective:To explore the effect of humanistic care on the quality of life and the negative emotion of patients with liver cancer operation. Methods: A total of 577 patients with liver cancer surgery were selected to participate in this study from January 2014 to December 2015 . From January 2014 to December 2014 , 288 cases of liver cancer patients were given routine nursing care as the control group. From January 2015 to December 2015, 289 patients were given humanistic care nursing as experimental group. The emotion of depression and anxiety were evaluated using Self-rating Depression Scale and Self-rating Anxiety Scale, the quality of life was evaluated with the core scale of life quality measurement system for cancer patients, while sleep quality with the Pittsburgh Sleep Quality Index. Results:Both depression and anxiety score of the subjects in experimental group were lower than those in the control group(P<0. 05). Physical function, cognitive function, social function, emotional function, role function, and overall health status of the subjects in the experimental group were higher than those in the con-trol group (P<0. 05), while pain, tiredness, nausea, vomiting, shortness of breath, insomnia, diarrhea, loss of appetite, constipation, economic difficulties and other score were statistically lower than those in the control group (P<0. 05). The subjects′quality of life of in the experimental group was better than those in the control group(P<0 . 05 ) , but the sleep index score was lower in the experimental group ( P<0 . 05 ) . Conclusion: Humanistic care nursing can effectively reduce the negative emotions of patients with liver cancer surgery, and thus to improve the quality of life.

Superoxide dismutase(SOD) was purified from Tegillarca granosa and the effects of tributyltin chloride(TBTCl) on the enzyme activity of the SOD were analyzed at different conditios. The results showed that the effects of TBTCl on SOD activity were more ohvious at room temperature than at four degrees centigrade. 58. 1 % of the enzyme activity was reserved after SOD was incubated with 200?g?mL-1 of TBTC1 for 72 hours at room temperature, while 90. 9% of the enzyme activity was reserved at four degrees centigrade. The enzyme activity of SOD was reduced with the increase of TBTC1 concentration and interaction time. TBTCl could reduce heat-resisting, acid-resisting and alkali-resisting abilities of SOD. With TBTCl concentration increase,SOD isozyme bands in the graph of polyacrylamide gel electrophoresis were getting weaker.

BACKGROUND:Immortalized cervical epithelial cels H8 can become cancerous under the induction of carcinogenic agent, and may cause cervical cancer when there is a cofactor interaction. However, there is stil a lack of effective intervention for female patients with precancerous lesions, and this treatment is blank in the clinic. OBJECTIVE: To explore the effects and mechanism of astragalus injection on apoptosis of human immortalizedcervical epithelial cels H8. METHODS: This study contained two groups: astragalus drug group and the blank control group. (1) Enzyme linked immunosorbent assay (ELISA) was used to detect DNA fragments of apoptotic H8 after astragalus injection. (2) Enzyme-labeling instrument was used to analyze the changes in caspase-3 and caspase-9 activities a fter astragalus injection. (3) Western blot assay was used to detect the protein expression changes of caspase-3, caspase-9 and PARP in H8 cels after astragalus injection. RESULTS AND CONCLUSION: (1) ELISA results showed that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, DNA fragments were gradualy increased with time prolonged in a time-dependent effect (P < 0.05). (2) Enzyme-labeling instrument demonstrated that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, caspase-3 and caspase-9 activities increased in a time-dependent manner (P < 0.05). (3) At 0, 6, 12 and 24 hours after 20 g/L astragalus injection, the expression of cleaved caspase-3 and cleaved caspase-9 were gradualy increased in H8 cels (P < 0.05). Cleaved PARP protein expression was gradualy decreased (P < 0.05). These findings indicate that astragalus injection could obviously induce H8 apoptosis, which may be associated with the upregulated protein expression of caspase-3 and caspase-9.

The simulator testing in vitro and computational simulation of the artificial knee joint wear are important methods to evaluate the wear performance of the prosthesis in vitro and to predict the clinical performance of knee joint products. Based on the method of literature search, this paper compares the mechanical and kinematic loading input curves carried out by Chinese scholars in recent years, standard curves, and Chinese measurement curves of two typical movements of gait. Data of vitro simulator test and computational simulation model are compared, summarized, and analyzed. The results show that the measured data of motion and load cannot be directly used as the loading conditions for the simulator wear test and computational simulation. The mechanics and kinematics data of Chinese people are different from the international standards. The domestic artificial knee joint in vitro simulator wear test methods are similar but the results of different test institutions are somewhat different. The computation wear prediction research is basically synchronized with foreign countries, but the problem that the calculated wear results are lower than that in vitro test is still unsolved. The artificial knee joint wear performance evaluation system based on Chinese knee joint mechanics and kinematics data is the forward direction of the research.

Objective:To investigate the characteristics of serum immunoglobulin and complement levels in centenarians in Hainan.Methods:Fasting venous blood samples from 969 centenarians in Hainan province were collected and from 364 seniors serving as the control group.Levels of serum immunoglobulin A, G, M and E, immunoglobulin light chain κ and λ, and complement C3 and C4 were measured.Serum immunoglobulin and complement levels were analyzed in subjects based on age, sex, ethnicity, diet and smoking.Results:Serum levels of immunoglobulin G and E, and immunoglobulin light chain κ were higher in the centenarian group than in the control group[15.9(13.8, 18.3)g/L vs.14.9(13.1, 16.9)g/L; 270.0(85.5, 851.0) vs.180.5 kU/L(58.0, 556.2)kU/L, 4.1(3.5, 4.9)g/L vs.4.0(3.1, 4.6)g/L, P<0.05]. Serum levels of immunoglobulin M, and complement C3 and C4 were lower in the centenarian group than in the control group[1.0(0.7, 1.4)g/L vs.1.1(0.8, 1.5)g/L, 1.0(0.9, 1.2) vs.1.1(1.0, 1.3), 0.2(0.2, 0.3) vs.0.3(0.2, 0.3), P<0.05]. Serum levels of immunoglobulin M and complement C3 were higher in female centenarians than in male centenarians[1.1(0.7, 1.4) vs.1.0(0.7, 1.3), 1.0(0.9, 1.1) vs.1.0(0.8, 1.1), P<0.05]. There were significant differences in immunoglobulin A, G and E, and immunoglobulin light chain κ and λ between centenarians of different ethnicities( P<0.01). Serum immunoglobulin M levels were higher in centenarians with a long history of milk-drinking than in those without the history[1.2(0.8, 1.5) vs.1.0(0.7, 1.4), P<0.01]. Conclusions:Serum levels of immunoglobulins and complements are different among centenarians of different ages, genders, ethnicities and diet habits.Factors such as advanced age, female gender, ethnic minority and milk drinking tend to promote the expression of immunoglobulins and complements.

Objective To investigate the expression of plasma microRNA-214 in patients with acute myocardial infarction (AMI) and its correlation with left ventricular remodeling (LVR). Methods A total of 158 AMI patients and 85 controls were selected from Hebei province Zhuozhou City Hospital. According to the left ventricular remodeling after PCI operation, AMI patients were divided into LVR group (n=41) and non LVR group (n=105). Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the plasma microRNA-214 levels, and the brain natriuretic peptide (BNP) and C-reactive protein (CRP) levels were detected by immunoassay. Pearson correlation analysis of plasma microRNA-214 levels and the correlation between CRP and BNP were analyzed in patients with LVR. The receiver operating characteristic (ROC) curve analysis of plasma microRNA-214, BNP and CRP were used to predict the efficacy of LVR in patients with AMI after PCI. Logistic regression model was used to analyze the relationship between the above indexes and LVR. Results Compared with the control group, left ventricular ejection fraction (LVEF) and left ventricular short axis shortening (FS) were significantly decreased in LVR group and non LVR group, and which were decreased more significantly in LVR group compared with those of non LVR group (P<0.05). The left ventricular end diastolic diameter (LVDD), left ventricular end diastolic volume (LVEDV) and left ventricular end systolic volume (LVESV) were significantly increased in LVR group and non LVR group, and which were increased more significantly in LVR group than those in non LVR group (P<0.05). The levels of plasma microRNA-214, BNP and CRP were significantly higher in LVR group than those in non LVR group and control group (P<0.05). Correlation analysis showed that microRNA-214, BNP and CRP were positively correlated in patients with LVR (r=0.684 and r=0.405, P<0.01). The area under ROC curve (AUC) and 95%CI predicted by plasma microRNA-214 and BNP were 0.824 (0.757-1.015) and 0.785 (0.721-0.864) in patients with AMI and LVR, which were higher than CRP [0.716 (0.645-0.837), P=0.0167]. The sensitivity and specificity of plasma microRNA-214 in predicting the occurrence of LVR were 72.6% and 86.2% in patients with AMI, respectively. Logistic regression model analysis showed that plasma microRNA-214 and BNP were risk factors of LVR. Conclusion High expression of plasma microRNA-214 is found in LVR patients, which is expected to be the biological indicator for predicting the occurrence of LVR after PCI in patients with AMI.

ObjectiveTo observe the effects of a modified version of constraint-induced movement therapy (CIMT) on upper-extremity function of children with hemiplegic cerebral palsy.Methods22 patients were divided randomly into treatment group (n=10) and control group (n=12). Cases of control group received routine rehabilitation therapy, otherwise, those of treatment group were treated with CIMT. It involved restraining the unimpaired extremity with a restraining-instrument, and having the child engage in activities with the involved extremity 4 hours a day for 2 months repeatedly. Before and after treatment, affected upper-extremity function of all children were assessed with the Fine Motor Function Measure (FMFM) scale.ResultsThe fine motor function of all children in two groups improved significantly(P<0.05), but effect of treatment group was better than that of control group (P<0.05).ConclusionThe treatment of CIMT can improve upper-extremity function of children with hemiplegic cerebral palsy.