BACKGROUND:Insulin-like growth factor-1(IGF-1)is an important cell factor which plays a special role in many disease treatments such as diabetes.The research of engineering bacteria fermentation production technology is great to IGF-1 industrialization and to clinical application.OBJECTIVE:To investigate the high cell-density fermentation and expression condition of recombinant human insulin-like growth factor-1(IGF-1) engineering bacteria.DESIGN,TIME AND SETTING:The enzyme,gene engineering,study was performed at the Biotechnological Laboratory of Zhejiang Shuren University from May 2007 to May 2008.MATERIALS:Recombinant E coli strain for IGF-1 expression as BL21 (DE3)/pET22a-IGF-1 was reserved in the Biotechnological Laboratory of Zhejiang Shuren University.Nutrient feed was composed of:glucose(300 g/L),peptone(40 g/L),yeast powder(10 g/L),Na2HPO4(280 mmol/L),Na2HPO4-2H2O2(120 mmol/L),MgSO4(10 mmol/L),and ampicillin(100 mg/L).METHODS:The strains were activated and then cultured in orbitaI shakers.Parameters such as types of media,isopropyl-β-D-thiogalactopyranoside(IPTG)concentration and induction time have been analyzed to explore optimaI fermentation conditions for expressing the recombinant protein.According to the optimal fermentation condition of orbitaI shakers.batch fermentation was carried on with 5 L-autocontroI fermentor.The process contained two stages:batch culture and fed-bafch through pH-stat solution.JPTG was added to jnduce the expression of protein in the middle and latter of the logarithmic growth phase for 4-βhours.MAIN OUTCOME MEASURES:Following described parameters were measured:fermentation and protein expression of the recombinant human IGF-1 engineering bacteria,cell concentration,cell dry weigh,objective protein,glucose concentration.RESULTS:Cells were cultured in 2×YT+5 g,L glucose medium,induced by 0.8 mmol/L IPTG for 5 hours.By controlling dissolved oxygen and by pH-stat feeding solution,high cell-density and high protein expression were achieved.Under the establishedconditions.50.1 g dry cell weight(DCW)/L could be obtained,and 5.25 g/L IGF-1 was achieved.CONCLUSION:The established high cell-density fermentative procedure of recombinant human IGF-1 engineering bacteda is the useful bases for further pudfication and large scale production of IGF-1.

Objective To observe the influence on early blood glucose by 3 L fluid replacementafter kidney transplantation. Methods Patients(60 cases) after kidney transplantation were randomly di-vidod into two groups.Group A used circular fluid replacement and Group B used 3L fluid replacement. Thelevel of blood glucose was detected before operation,after operation ,once every 8 hours at the first day,thenonce a day for the following six days. Results The level of blood glucose with 3 L fluid replacement waslower than that with circular fluid replacement,especially from the fast day to the fourth day after operation(P < 0.05). Conclusions The method with 3 L fluid replacement surpasses obviously circular fluid re-placement in blood glucose control after kidney transplantation.

Objective To investigate the relationship between serum levels of homocysteine (HCy) and adiponectin (APN) in patients with type 2 diabetes mellitus(T2DM).Methods 65 patients with T2DM (diabetes mellitus group) and 25 healthy controls (control group) matched in the age and sex were recruited in the study.Serum HCy,APN,fasting plasma glucose (FPG),fasting insulin (FINS),total cholesterol (TC) and triglyceride (TG) were simultaneously measured.The homeostatic model assessment for insulin resistance(HOMA-IR) was calculated according to FPG and FINS.All the serum indicators were compared between the two groups.Results Serum level of HCy in T2DM group was (15.74 ± 2.76) μmol/L,which was significantly higher than (6.98 ± 1.94) μmol/L in the healthy control group (t =16.88,P ＜ 0.01).The serum level of APN in T2DM group was (8.14 ± 2.70) mg/L,which was significantly lower than (16.10 ± 1.93)mg/L in the healthy control group (t =13.44,P ＜ 0.01).Serum levels of FPG,HOMA-IR,TC,TG in T2DM group were significantly higher than those in the healthy control group (t =10.62,17.49,6.30,7.52,P ＜ 0.05).Serum level of APN in HCy ≥ 15μmol/L group was significantly decreased compared with HCy ＜ 15μmol/L group.Serum level of HCy was negatively correlated with APN in T2DM group after the influence of FPG,HOMA-IR,TG,TC were corrected in the partial correlation analysis.Conclusion In T2DM group,serum level of HCy was increased,but serum level of APN was decreased,serum HCy was negatively correlated with APN,higher serum level of HCy and lower serum level of APN are related with the process of insulin resistance and T2DM.

Objective To investigate the changes of serum homocysteine and it s relationship with oxidative stress and diabetic microangiopathy.Methods Eighty health participants were recruited as control 100 type 2 diabetis patients without diabetic microangiopathy were recruited as DM group,100 type 2 diabetis patients with diabetic nephropathy were recruited as DN group,and 100 type 2 diabetis patients with diabetic retinopathy were recruited as DR group.Their serum levels of homocysteine,malonaldehyde(MDA),superoxide dismutase (SOD)and glutathione(GSH)were measured.Results The homocysteine was(98.86 ± 21.46),(198.95 ±19.35),(138.65 ± 15.25)ng/L in the DM,DN and DR group respectively,which were signigicantly higher than that of(62.48 ± 15.36)ng/L in the control group(F =7.95,P ＜ 0.01).MDA was(17.49 ± 1.64),(22.47 ± 1.86)and(22.47 ± 1.86)mmol/L,which was significantly higher than that of(11.86 ± 0.48)mmol/L in the control group(F =6.89,P ＜0.01).The homocysteine and MDA in the DN and DR group were both significantly higher than those in the DM group(P ＜ 0.01).The SOD and GSH was(107.80 ± 15.62)mg/L and(179.26 ± 25.8)mg/L in the DM group,(79.86 ± 14.63)mg/L and(143.36 ± 21.75)mg/L in the DN group,(89.34 ± 12.75)mg/L and(156.96 ± 19.35)mg/L in the DR group,which were significantly higher than those of(128.32 ± 19.21)mg/L and(237.38 ± 27.31)mg/L in the control group(F =7.89 and 8.76 respectively,P＜0.01).The SOD and GSH in the DN and DR group were both significantly lower than those in the DM group(P ＜ 0.01),and the DN group was significantly lower than the DR group(P ＜ 0.01)Serum homocysteine was positively correlated with MDA(r =0.79,P ＜ 0.05),and negtively correlated with SOD and GSH(r =-0.71 and-0.78,P ＜0.01).Conclusion Diabetic microangiopathy patients have higher serum homocysteine level and severe oxidative stress.Oxidative stress were related to higher serum homocysteine level.The higher serum homocysteine level and oxidative stress might play an important role in development of diabetic Microangiopathy

Objective To investigate the relationship between intima-media thickness(IMT)and pulse wave velocity(PWV)in patients with type 2 diabetes mellitus.Methods 100 cases with type 2 diabetes were randomly divided into:hydrochlorothiazide group of 35 cases,valsartan group of 35 cases,valsartan and hydrochlorothiazide combined treatment group(combined group)30 cases.All the patients were treated for 16 weeks,systolic blood pressure (SBP),diastolic blood pressure(DBP),24h urinary albumin,blood urea nitrogen(BUN),carotid IMT,PWV,fasting plasma glucose(FPG)were measured before and after treatment in each group.Results The SBP,DBP,24h urinary albumin before treatment compared with after treatment was significant differences in three groups(t =2.7875,2.755,2.733,2.645,all P ＜ 0.05); IMT associated with SBP(r =0.669,P =0.013),pulse pressure(r =0.581,P =0.015),FPG(r =0.337,P =0.018);PWV associated with SBP(r =0.759,P =0.001),pulse pressure(r =0.707,P=0.002),FPG(r =0.679,P =0.016).Conclusion The non-invasive indicators of PWV can be used as aortic compliance.The PWV measurement of its vascular complications has important clinical value.

Objective To identify risk factors and preventive measures of nosoconial infections in patients with non-Hodgkin lymphoma (NHL). Methods Clinical data of 65 NHL patients admitted from January to December 2007 were retrospectively analyzed. Results According to WHO classification (2001), 58 patients were with B-cell lymphoma, 7 were with T-cell lymphorna. All patients received CHOP regimen as initial chemotherapy and 23 of them were with nosecomial infections. Logistic regression analysis demonstrated that age, length of stay, pathological type, bone marrow involvement, levels of serum lactate dehydrogenase (LDH), beta2-microglobulin and invasive treatment were identified as risk factors of nosocomial infections. Respiratory tract infections and infections with gram-negative microorganisms were the most popular. Conclusion High nosocomial infection rate is found in NHL patients, and control of risk factors may effectively prevent nosocomial infections in NHL patients.

Objective To explore the application effect of the adult education theory in the nursing round. Methods 340 trainee nurses, who were trained at our surgical department between June 2008 and December 2009, were randomly divided into the experimental group(178 cases)and the control group(162cases). The adult education theory was adopted in the experimental group, whereas the traditional education method was used in the control group, a questionnaire was carried out after the nursing visit. Results The experimental group was more satisfied with the round visit than the control group. Conclusions The application of the adult education theory in the round visit is beneficial not only to the students' selection of the topics of nursing visit, but also to the improvement of the students' understanding and communicating capacity, the students' studying interests and the students' creative thinking, finally the degree of satisfaction to the round visit is increased by the adoption of this theory.

Objective To study the changes of endothelin,nitric oxide and vascular endothelial growth factor level in patients with type 2 diabetic retinopathy (DR). Methods Eighty diabetes patients (53 with diabetic retinopathy and 27 without). Another 30 healthy volunteers were recruited as control. Plasma ET and VEGF levels were measured by enzyme-linked immunosorbent assay. NO levels were measured by nitrate reductase method. Results Plasma levels of ET were higher in patients with type 2 diabetes with DR (DR)(80. 68 ± 13.57) mg/L than (65. 33 ± 11.24) mg/L, (45.25 ±9. 06) mg/L, in control and in type 2 diabetes patients without DR (NDR) (Ps ＜ 0, 01 ). Plasna levels of NO in DR group (69. 82 ± 14. 89) μmol/L were higher than (37. 85 ±-9. 11 ) μmol'L, in control group,but lower than (77.52 ±± 18.56) μmol/L in NDR group (Ps ＜ 0. 05 ). Plasma levels of VEGF ( 110. 52 ± 25.65 ) μg/L in DR were significantly increased compared with control (82.42 ± 18. 47 ) μg/L, and NDR(97.55 ±25.61)μg/L, (Ps ＜0.05).Conclusion ET, NO and VEGF may be involved in the pathogenesis of type 2 diabetic retinopathy.

OBJECTIVE To isolate bacteria from three second-class hospitals in Zhenjiang Jiangsu Provine,to detect their drug(-resistance) to antibacterial(agents) and analyzed the results in order to find out pathogen distribution and drug(-resistance) trend of the common causative organism.METHODS We isolated 1661 strains of bacteria from clinical samples between 2002 and 2004,and analyze their drug-resistance.RESULTS The number of Gram-positive(bacteria) was 634,accounting for 38.2%,from them there were 312 strains of coagulase-negative Staphylococcus epidermidis,and 148(coagulase)-positive S.aureus.Drug-fast of two above staphylococci was similar.The(resistance) rate to penicillin and to oxacillin surpassed 85%,that to erythromycins exceeded 75%,to quinolones and cephalosporins surpassed 30% and 35%,respectively.The resistance rate to(vancomycin) was zero.Gram-(negative) bacteria were 1 027 strains,accounted for 61.8%.(According) to quantity order,the first was Enterobacter(249 strains);then were Klebsiella(226 strains);Escherichia(167 strains);Pseudomonas aeruginosa(136 strains);and Proteus(62 strains).The resistant rate to semisyhthesized penicillins among the first five Gram-(negative) bacteria species,was the most,especially the Proteus.CONCLUSIONS Clinical doctors,the(laboratorians) and detecting infection department in hospital should markedly pay attention to drug-resistant(bacterium) detection,pathogen mutation and drug-resistance trend,in(order) to reasonably use antibacterial(agents).

Objective To explore clinical application value of Traditional Chinese Medicine nursing technology to decrease postoperative complication after breast cancer surgery.Methods Randomized controlled trial,large sample size,multicenter study design were adopted,and 200 patients who met inclusion criteria were randomly divided into the control group and the treatment group (100 cases in each group).The control group received routine nursing methods after breast cancer surgery.The treatment group received routine nursing methods as well as Traditional Chinese Medicine nursing intervention,such as auricular application pressure,acupoint sticking,meridian moxibustion.Data of arm circumference,Athens insomnia scale(AIS),symptom in affected arm were collected a week before and after surgery,and range of motion of shoulder joint was evaluated 3 months after surgery to compare postoperative subcutaneous effusion,skin flap necrosis,limb edema,sleep disorder and occurrence of shoulder joint dysfunction in two groups.Results The cases developed postoperative subcutaneous effusion,skin flap necrosis,limb edema were different in two groups,but the differences were not statistically significant(P＞0.05).There were significant differences between two groups in affected arm pain,swelling,skin tension,sleep,and shoulder joint dysfunction after 3 months(P＜0.05).Conclusion Traditional Chinese Medicine nursing technology can effectively decrease postoperative complication after breast cancer surgery.