The Use of Escherichia coli for Recombinant Human Insulin Production

Insulin is the essential hormone produced by the pancreas. which is accountable for sanctioning glucose we acquire from our food sources to be deposited in our body cells. Without insulin, our bodies cannot control blood sugar levels, so insulin is a vital hormone for survival. A diabetic person either does not produce insulin or is resilient to it for a multiple reasons. Because of this, they need insulin injections to process glucose. It has become stress-free for patients around the world to acquire insulin with the production of recombinant human insulin produced by Escherichia coli . This short review will provide an overview of the steps engaged in constructing recombinant human insulin utilizing the K12 strain of E. coli along with the prominence of recombinant insulin and why E. coli is most commonly used for insulin production.

The Effect of DPP4 Inhibitor on Glycemic Variability in Patients with Type 2 Diabetes treated with twice-daily Premixed Human Insulin / Journal of the ASEAN Federation of Endocrine Societies

Objective@#To evaluate the effect of adding DPP4 inhibitor (DPP4-i) on glycemic variability (GV) in patients with type 2 diabetes mellitus (T2DM) treated with premixed human insulin (MHI).@*Methodology@#We conducted a prospective study in patients with T2DM on twice-daily MHI with or without metformin therapy. Blinded continuous glucose monitoring was performed at baseline and following 6 weeks of Vildagliptin therapy.@*Results@#Twelve patients with mean (SD) age of 55.8 (13.1) years and duration of disease of 14.0 (6.6) years were recruited. The addition of Vildagliptin significantly reduced GV indices (mmol/L): SD from 2.73 (IQR 2.12-3.66) to 2.11 (1.76-2.55), p=0.015; mean amplitude of glycemic excursions (MAGE) 6.94(2.61) to 5.72 (1.87), p=0.018 and CV 34.05 (8.76) to 28.19 (5.36), p=0.010. In addition, % time in range (3.9-10 mmol/l) improved from 61.17 (20.50) to 79.67 (15.33)%, p=0.001; % time above range reduced from 32.92 (23.99) to 18.50 (15.62)%, p=0.016; with reduction in AUC for hyperglycemia from 1.24 (1.31) to 0.47 (0.71) mmol/day, p=0.015. Hypoglycemic events were infrequent and the reduction in time below range and AUC for hypoglycemia did not reach statistical significance.@*Conclusion@#The addition of DPP4-I to commonly prescribed twice-daily MHI in patients with T2DM improves GV and warrants further exploration.

Cost-effectiveness Analysis of Insulin Analogues and Human Insulin in the Treatment of Type 2 Diabetes Mellitus Based on Net-benefit Regression Framework / 中国药房

OBJECTIVE：To compare the cost-effectiveness of insulin analogues and human insulin in the treatment of type 2 diabetes mellitus ，to reduce economic burden of patients ，and to provide new ideas for non-randomized pharmacoeconomic research. METHODS ：Retrospective data of a Third-grade Class-A hospital were adopted and 533 patients with type 2 diabetes were taken as object to balance the difference between groups through propensity score matching ，and construct net-benefit regression framework so as to analyze the cost-effectiveness of insulin analogues and human insulin in the treatment of type 2 diabetes mellitus. RESULTS ：There was a positive relationship between the efficacy and cost of the two therapeutic schemes ，i.e. insulin analogues were more effective than human insulin ，and the difference of total effective rate between the two schemes was 14.5％. The corresponding treatment cost of insulin analogues was higher than that of human insulin ，and the average total cost difference was 960.3 yuan. The cost of insulin analogues was 66.23 yuan more than that of human insulin for each additional unit of clinical effect （total effective rate ）. Results of net-benefit regression analysis showed that critical value of willingness to pay of therapy plan was 16 947.5 yuan. CONCLUSIONS ：When the willingness to pay is more than 16 947.5 yuan，the insulin analogue scheme is suitable ；on the contrary ，the human insulin analogue scheme is suitable. The propensity scoring technology used for the net-benefit regression analysis can broaden the source of avaliable data for non-randomized pharmacoeconomic evaluation.

Effectiveness of insulin analogs compared with human insulins in pregnant women with diabetes mellitus: systematic review and meta-analysis / Efetividade dos análogos da insulina comparados às insulinas humanas em gestantes com diabetes mellitus: Revisão sistemática com metanálise

Abstract Diabetes during pregnancy has been linked to unfavorable maternal-fetal outcomes. Human insulins are the first drug of choice because of the proven safety in their use. However, there are still questions about the use of insulin analogs during pregnancy. The objective of the present study was to determine the effectiveness of insulin analogs compared withhuman insulin in the treatment of pregnant women with diabetes througha systematic review withmeta-analysis. The search comprised the period since the inception of each database until July 2017, and the following databases were used:MEDLINE, CINAHL, EMBASE, ISIWeb of Science, LILACS, Scopus, SIGLE andGoogle Scholar.We have selected 29 original articles: 11 were randomized clinical trials and 18 were observational studies.We have explored data from 6,382 participants. All of the articles were classified as having an intermediate to high risk of bias. The variable that showed favorable results for the use of insulin analogs was gestational age, with a mean difference of - 0.26 (95 % confidence interval [CI]: 0.03-0.49; p = 0.02), but with significant heterogeneity (Higgins test [I2] = 38%; chi-squared test [χ2] = 16.24; degree of freedom [DF] = 10; p = 0.09). This result, in the clinical practice, does not compromise the fetal well-being, since all babies were born at term. There was publication bias in the gestational age and neonatal weight variables. To date, the evidence analyzed has a moderate-to-high risk of bias and does not allow the conclusion that insulin analogs are more effective when compared with human insulin to treat diabetic pregnant women.

Resumo Diabetes durante a gestação tem sido relacionado a desfechos materno-fetais desfavoráveis. As insulinas humanas são a primeira escolha medicamentosa, devido à comprovada segurança no seu uso. Entretanto, ainda há questionamentos sobre o uso dos análogos da insulina na gestação. O objetivo do presente estudo foi determinar a efetividade dos análogos da insulina comparados às insulinas humanas no tratamento de gestantes com diabetes por meio de uma revisão sistemática com metanálise. A busca compreendeu desde o início de cada base de dados até julho de 2017, e foi realizada nos seguintes bancos de dados: MEDLINE, CINAHL, EMBASE, ISI Web of Science, LILACS, Scopus, SIGLE e Google Scholar. Selecionamos 29 artigos originais, sendo 11 ensaios clínicos randomizados e 18 estudos observacionais. Exploramos dados de 6.382 participantes. Todos os artigos foram classificados como sendo de intermediário a alto risco de viés. A variável que demonstrou resultado favorável ao uso dos análogos da insulina foi idade gestacional, com uma diferençamédia de - 0.26 (95% índice de confiança [IC]: 0.03-0.49; p = 0.02), porém com heterogeneidade significativa (teste de Higgins [I2] = 38%; teste do qui quadrado [χ2] =16.24; graus de liberdade [GL] =10; p = 0.09). Esse resultado, na prática clínica, não compromete o bem-estar fetal, uma vez que todos os bebês nasceram a termo. Houve viés de publicação nas variáveis idade gestacional e peso neonatal. Até o momento, as evidências analisadas possuem um risco de viés moderado a elevado e não permitem concluir que os análogos da insulina sejam mais efetivos em comparação às insulinas humanas para tratar gestantes diabéticas.

Effects of glargine insulin on glycemic control in patients with diabetes mellitus type II undergoing off-pump coronary artery bypass graft

Background: The prevalence of diabetes mellitus in patients requiring coronary artery bypass grafting (CABG) is noticeably high (20%–30%). These patients have inferior perioperative outcome, reduced long-term survival, and high risk of recurrent episodes of angina. To improve perioperative outcome surgical unit defined satisfactory glycemic control is desired during this period. Hence, the aim of our study is to compare the efficacy of glargine insulin combination with continuous human insulin infusion for perioperative glycemic control in patients with diabetes undergoing CABG. Materials and Methods: Fifty Patients, who were posted for off-pump CABG with diabetes mellitus type II, were randomized in two group, Group I normal saline + human insulin infusion during the perioperative period, Group II (glargine group): Glargine + human insulin infusion during perioperative period. Results: During surgery and in the postoperative period, random blood sugar and human insulin requirement are significantly higher in control group than glargine group. Other infection, step-up antibiotics, intensive care unit (ICU) stay, and hospital stay were significantly higher in control groups in postoperative period. Conclusion: Our study results suggest that glargine effectively manages blood glucose level with significantly greater control over postoperative morbidity.

Thirty years of experience with recombinant human insulin-from familiarity to reliance / 中国糖尿病杂志

Recombinant human insulin,a milestone of diabetes treatment,has been put into clinical practice for over 30 years since 1980s.Its efficacy,safety and cost-effectiveness have been confirmed by clinical practice and clinical studies.Based on evidences,recombinant human insulin,among many antidiabetic agents,still remains one of the core hypoglycemic drugs recommended by authoritative guidelines for the prevention and treatment of diabetes.This review summarized the development,clinical efficacy,safety profiles and guideline recommendations of recombinant human insulin to lay the cornerstone of individualized insulin therapy.

Cost-minimization Analysis of Four Kinds of Premixed Insulin in the Therapy for Newly Diagnosed Type 2 Diabetes / 中国药师

Objective:To evaluate the short-term economic effects of four kinds of premixed insulin in newly diagnosed type 2 dia-betes mellitus. Methods:A total of 120 newly diagnosed patients with type 2 diabetes mellitus were divided into four groups according to the kind of premixed insulin, group A was treated with insulin aspart 30 injection, group B was treated with insulin lispro 25 injec-tion, group C was treated with isophane protamine biosynthetic human insulin injection and group D was treated with protamine zinc re-combinant human insulin injection. The course of treatment was three months. The therapy efficacy was assessed by the remission rate in three months. The short-term economic effect was evaluated by the cost-minimization analysis method. Results:The remission rate of group A, B, C and D respectively was 48. 39%, 48. 28%, 51. 61% and 51. 72% without significant difference (P>0. 05). The average cost per person of the four groups was 1195. 52, 1202. 41, 1220. 69 and 1258. 84 yuan, and the average medicine cost per person was 750. 52, 689. 41, 754. 69 and 764. 34 yuan, respectively. There was no significant difference in cost among the four groups (P >0. 05). Conclusion:All the four kinds of premixed insulin can be used for the starting treatment with the similar total cost, and in relative terms, aspart 30 injection and insulin lispro 25 injection are better for the initial treatment of diabetes.

Experimental Study on Effect of Drynaria Combining with Tissue Engineered Cartilage to Promote Cartilage Regeneration / 现代生物医学进展

Objective:To evaluate the effect of rhizome drynaria combined with tissue engineering cartilage on cartilage regeneration in experimental rabbits with cartilage defects.Methods:The hIGF-1 gene was transfected into MSCs by using the method of isola tion,purification and recombination of transgenic stem cells.The MSCs were transplanted into rabbit bone marrow mesenchymal stem cells (MSCs) in vitro.The cells were further amplified and mixed with acellular dermal matrix (ADM) to construct tissue engineered cartilage.Twenty-four New Zealand white rabbits,aged 6 months,were randomly divided into 4 groups (A,B,C and D).six rabbits in each group.Group A and C were transplanted with autologous cartilage.Group B and D were transplanted with modified cells.Group C and D group were fed with 40％ Drynaria Decoction,150ml/d for 4 weeks.Animals were sacrificed at 12 weeks postoperatively,and articular cartilage defects were isolated.Cartilage defect samples were embedded in paraffin blocks and stained with hematoxylin and eosin (H&E).Cartilage regeneration was evaluated by gross morphology,including sclerotic shape,color,contour and homogeneity.The quality of regenerated cartilage was assessed by histological scoring.Toluidine blue staining was used to evaluate the occurrence of chondrogenic glycosaminoglycans (GAG).Results:Compared with group B,the cartilage coverage,the color of new bone marrow,the edge of defect and the surface roughness of group C and D were significantly improved (P＜0.05);the cartilage surface score of regenerated cartilage was significantly improved P＜0.05).Groups C and D had better matrix,cell distribution and surface index than the other groups.And had a thick like hyaline cartilage tissue,with the normal glycosaminoglycan production.It is indicated that drynaria combined with tissue engineering cartilage can reduce cartilage defects by regenerating hyaline cartilage.Conclusion:Cartilage combined with drynariae can significantly improve the quality of cartilage defect repair in rabbit knee joint,and provide an important theoretical basis for clinical treatment of cartilage lesions.

Meta-analysis of insulin aspart versus regular human insulin in diabetes treatment in China / 中国糖尿病杂志

Objective To compare the efficacy and safety of insulin aspart (IAsp) versus regular human insulin (RHI) used in basal bolus (BB) or continuous subcutaneous insulin infusion (CSII) regimen in patients with diabetes mellitus in Chinese population.Methods We searched MEDLINE (via OVID),the Cochrane Library,Embase,PubMed,CBM,China National Knowledge Infrastructure (CNKI),and Wanfang databases from the beginning of the databases to November,2015.The randomized controlled trials comparing IAsp and RHI in DM were searched.The meta-analysis in newly diagnosed DM and treated DM was performed by RevMan 5.3.Results A total of 40 trials were included in this study,with 1087 newly diagnosed and 2395 treated DM patients.In both groups,IAsp was better in lowering 2 hours postprandial plasma glucose (2 hPG) after breakfast,lunch and dinner (For newly diagnosed population MD=-1.22,-1.70,-1.44;95%CI:-1.79~-0.64,-2.77~-0.63,-2.12~-0.75;for treated population MD=-1.19,-1.14,-1.03;95%CI:-1.39~-0.98,-1.27~-1.02,-1.25~-0.81) and fasting plasma glucose (For newly diagnosed population MD=-0.55,95%CI:-0.97~-0.12;for treated population MD=-0.24,95%CI:-0.44~-0.03) when compared with RHI.IAsp could shorten the time to achieve blood glucose target(For newly diagnosed population MD=-1.58,95%CI:-1.74~-1.42;for treated population MD=-1.73,95%CI:-2.10~-1.36) and reduce the risk of hypoglycaemia compared with RHI (For newly diagnosed population RR=0.33,95%CI:0.19~0.56;for treated population RR=0.44,95%CI:0.32~0.61) (P<0.05).Moreover,the total daily insulin dose was lower in IAsp therapy than in RHI therapy when using CSII regimen.Conclusion In Chinese DM patients,IAsp shows advantages in controlling 2 hPG,shortening the time to achieve blood glucose target and lowering the risk of hypoglycaemia when compared with RHI.

Macromolecular confinement of therapeutic protein in polymeric particles for controlled release: insulin as a case study

ABSTRACT Sustained release systems for therapeutic proteins have been widely studied targeting to improve the action of these drugs. Molecular entrapping of proteins is particularly challenging due to their conformational instability. We have developed a micro-structured poly-epsilon-caprolactone (PCL) particle system loaded with human insulin using a simple double-emulsion w/o/w method followed by solvent evaporation method. This formulation is comprised by spheric-shaped microparticles with average size of 10 micrometers. In vitro release showed a biphasic behavior such as a rapid release with about 50% of drug delivered within 2 hours and a sustained phase for up to 48 h. The subcutaneous administration of microencapsulated insulin showed a biphasic effect on glycemia in streptozotocin-induced diabetic mice, compatible with short and intermediate-acting behaviors, with first transition peak at about 2 h and the second phase exerting effect for up to 48h after s.c. administration. This study reveals that a simplified double-emulsion system results in biocompatible human-insulin-loaded PCL microparticles that might be used for further development of optimized sustained release formulations of insulin to be used in the restoration of hormonal levels.

Cost-effectiveness analysis of insulin Aspart30 versus premixed human insulin 30/70 in patients with type 2 diabetes in China / 中国药学杂志

OBJECTIVE: To evaluate the cost-effectiveness of insulin aspart30(BIAsp 30) versus premixed human insulin 30/70 (BHI 30) in patients with type 2 diabetes (T2DM) in China so as to provide a reference for relevant decision of drug selection and drug pricing. METHODS: Based on a secondary document study method and literature review both at home and abroad, a summary and screening were made on the safety and effectiveness of BIAsp 30 versus BHI 30 for the treatment among insulin-naive people with T2DM. A Meta-analysis was performed to assess the effectiveness and safety of the selected data by using software RevMan5.2. Then combine the price from the National Development and Reform Commission, National Health and Family Planning Commission to perform cost-effectiveness analysis from a social prospective. Sensitivity analysis and publication bias were conducted. RESULTS: Treatment with BIAsp 30 is associated with more cost than BHI 30 (¥1 665.44 vs ¥1 383.86), and the effectiveness data of BIAsp 30 was higher than BHI 30 (2.46 vs 1.83). Compared with BHI 30, BIAsp 30 is more efficient in pharmacoeconomic cost-effectiveness analysis, and sensitivity analyses demonstrated robustness of the results. CONCLUSION: BIAsp 30 is better than BHI 30 in terms of the economy, but it still needs further research.

Clinical efficacy of short term intensive insulin pump therapy and biosynthetic human insulin therapy and its effect on maternal and neonatal outcomes in gestational diabetes mellitus / 中国生化药物杂志

Objective To investigate the clinical effect of insulin on gestational diabetes mellitus(GDM).Methods GDM patients were randomly divided into group A and group B,group B were treated with short term intensive insulin pump therapy,and group A were treated with insulin pump in short term combined with biosynthetic human insulin therapy.Results The body mass index and diabetes family history rates pre-pregnancy in group B were significantly higher than group A(P<0.05).Logistic analysis showed that family history of BMI pre-pregnancy and diabetes family history were independent risk factors for GDM drug treatment(P<0.05).Compared with the group B,the improvement of blood glucose levels and the incidence of complications in patients with group A were better than the former(P<0.05).Conclusion Rational application of insulin can improve the efficacy of clinical treatment of GDM, reduce the incidence of complications, with high safety.

Comparison of efficacy and safety between biphasic insulin aspart 50 and biphasic human insulin 50：A randomized crossover trial / 中华内分泌代谢杂志

Objective To investigate the efficacy of biphasic insulin aspart 50(BIAsp50)twice daily(bid) versusbiphasichumaninsulin50(BHI50)(bid)plusmetforminonbloodglucosecontrolfollowingastandardmealtest in Chinese patients with type 2 diabetes mellitus(T2DM). Methods A randomized, open-label, 2-sequence, crossover trial for two 4-week treatment periods was conducted in 14 Chines institutes. Eligible subjects inadequately controlled with BHI50(bid)plus metformin were randomized to two sequences in a 1 : 1 ratio(A:BIAsp50-BHI50, B:BHI50-BIAsp50 ) . Standard meal tests were performed at baseline and the ends of two periods within 4 weeks. Primary endpoint was 2h postprandial plasma glucose ( PPG) increment following standard meal test, with insulin dose standardized at 0. 3 IU/kg. Results A total of 161 subjects were randomized into two sequences(81 to sequence A, and 80 to sequence B) and finally analysed. After 4 weeks of treatment, mean 2h PPG increment with BIAsp50 was lower than that with BHI50 [ treatment difference of BIAsp50 vs BHI50: -1. 12 mmol/L ( 95% CI-1. 66,-0. 58), P<0. 01], suggesting superiority of BIAsp50 over BHI50. Incremental area under the curve for PPG(0-2 h)with BIAsp50 was lower than that with BHI50 [treatment difference:-38. 8 mmol·L-1·min-1(95%CI-77. 3,-0. 26), P=0. 049], as was the mean 2h PPG [treatment difference:-0. 58 mmol/L(95% CI -1. 13,-0. 03), P=0. 040]. The FPG value with BIAsp50 was higher than that with BHI50 [treatment difference:0. 52 mmol/L(95%CI 0. 18, 0. 86), P=0. 003]. The rate of nocturnal hypoglycemia with BIAsp50 was lower than that with BHI50(1. 13 vs 2. 86 events per subject year, P<0. 01). Conclusion In patients with T2DM inadequately controlled with BHI50 plus metformin, BIAsp50 was proven to be well-tolerated with improved postprandial glucose control compared with BHI50.

Effect of insulin injection and protamine biosynthetic human insulin injection on basal insulin level in pregnant women with diabetes mellitus / 中国生化药物杂志

Objective To explore the effect of insulin injection and protamine biosynthetic human insulin injection on basal insulin level in pregnant women with diabetes mellitus.Methods Retrospective analysis of 89 cases of pregnant women with diabetes mellitus from January 2013 to May 2016 in department of obstetrics and gynecology,tianjin red bridge hospital,the patients were divided into group A (n=38 cases) and group B (n=51 cases),the group A treatment with insulin injection,the group B treatment with protamine biosynthetic human insulin injection,compare the two groups of patients before and after treatment of three meals a day rate of blood glucose compliance, treatment compliance and satisfaction.Results Before treatment, there was no significant difference in the blood glucose compliance rate between the two groups before and after treatment;After treatment,the blood glucose compliance rate of two groups was significantly higher than before treatment ( P<0.05 ) , there was no significant difference in the compliance rate of fasting blood glucose between two groups,the compliance rate of blood glucose before dinner in group A was 81.58%,significantly higher than that in group B 60.78%(P<0.05).Conclusion Both insulin injection and protamine biosynthetic human insulin injection can maintain the basic insulin levels of pregnant women with diabetes mellitus,insulin injection can better control the blood glucose levels before dinner,with higher compliance and satisfaction.The compliance rate and satisfaction rate of pregnant women in group A were 97.37% and 97.37%,which were significantly higher than those in group B 82.35% and 80.39%(P<0.05).

A novel rapid - acting oral inhalation human insulin for diabetes mellitus: Afrezza.

Afrezza is rapid-acting oral inhalation insulin that is administered at the beginning of each meal. The U.S Food and Drug Administration has approved Afrezza (insulin human) inhalation powder, a rapid-acting inhaled insulin to improve glycemic control in adults ≥18 years of age with Type 1 or Type 2 diabetes mellitus (T1DM or T2DM). Afrezza must be used in combination with long-acting insulin in patients with T1DM. Afrezza may be used with either oral anti-diabetic drugs or basal insulin in patients with T2DM. Afrezza should be administered via oral inhalation using Afrezza inhaler. Dosage adjustment is needed when switching from injection insulin to oral inhalation Afrezza. It is contraindicated in individuals with chronic lung disease and smokers because of the risk of the acute bronchospasm. Before initiating, Afrezza, a complete medical history, physical examination and spirometry (forced expiratory volume 1 sec) results is required in all individuals to identify the potential lung disease. Common adverse reactions in individuals treated with Afrezza include hypoglycemia, cough, throat pain or irritation, headache, and diarrhea.

Inhibitory Effect of siRNA Expression Vector Inhibiting IGF2 Gene on the Proliferation of Hepatoma Cell Line Huh-7 / 中国药房

OBJECTIVE:To investigate the inhibitory effect of siRNA expression vector inhibiting human insulin-like growth factor 2(IGF2)gene on the proliferation of hepatoma cell line Huh-7. METHODS:siRNA expression vector pGL3-hAFP-hTERT-siRNA3(“siRNA3”)which inhibited IGF2 gene by dual promoter regulation of recombinant human alpha-foetoprotein(hAFP)and human telomerase reverse transcriptase(hTERT)was transfected into the Huh-7 cell and normal hepatocyte L-02,and then a nega-tive control group(vector pGL3-hAFP-hTERT)and a blank control group were set up. IGF2 mRNA expression was detected by re-al-time fluorescent quantitative polymerase chain reaction 48 h after transfection into the cells in all groups;the activity of the cells by the microplate reader 0,24,48 and 72 h thereafter;and the cell cycle and apoptosis by the flow cytometer 48 h thereafter,and the changes in the protein levels of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in the cell were detected by Western blot. RESULTS:Compared with the negative control group and blank control group,IGF2 mRNA expression in the Huh-7 cell transfected with siRNA3 was obviously weaker;at 48 and 72 h after transfection,the activity of Huh-7 cell signigicantly reduced, Huh-7 cells at G1 phase obviously increased and those at S phase markedly decreased;the occurrence of early,late and total apopto-sis in Huh-7 cells apparently increased,and the protein expression of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in cells significantly weakened,with statistically significance(P0.05). CONCLUSIONS:siRNA which inhibited IGF2 gene by dual promoter regulation of recombinant hAFP and hTERT can specially inhibit IGF2 gene expression and the prolifer-ation of Huh-7 cells,which may be involved with down-regulated protein expression of cell proliferation-associated gene PCNA, cell cycle control-associated genes Cyclin E2,Cyclin D2 and Cdc2 and apoptosis regulation-associated gene Bcl-2 as a result of down-regulated IGF2 mRNA expression and protein expres-sion.

"Construction and preliminary validation of a new ""Controllable"" recombinant adenovirus vector carrying human insulin gene" / 实用医学杂志

Objective To design and construct a controlled adenovirus vector in degradating by itself after induction for solving the problem of stimulating host immune and producing replication adenovirus and providing a secure exogenous gene vectors for clinical practice. Methods Based on the traditional adenovirus vector AdEasyTM system, we inserted the Cre gene which belongs to Cre-LoxP system into the downstream of Tet-On inducible expression system. Two LoxP sites were inserted into two sides of the shuttle plasmid′s right arm genome. Then, the full-length human insulin gene was inserted HindIII enzyme site. After the recombinant adenovirus infected the rat bone marrow-derived mesenchymal stem cells , fluorescent protein expression and insulin secretion were detected before or after induction by Dox. Results A new controlled recombinant adenovirus vector carrying human insulin gene was constructed successfully, and was named AdEasyN/INS. After the transfection of this new vector into QBI-293A cells and rat bone marrow mesenchymal stem cells , green fluorescent protein could be observed. After induction by Dox, both of the ratio of fluorescent cells/total cell and the levels of insulin significantly decreased. Conclusion Construction and preliminary validation of a controlledrecombinant adenovirus vector carrying human insulin gene is constructed successfully , it could infect rat bone marrow mesenchymal stem cells, and degradate by itself after Dox induction, realize the controllability of exogenous gene carrier.

Human insulin inhibits apoptosis and promotes proliferation of rat liver cell line BRL-3 A / 基础医学与临床

Objective_To study the effect of human insulin on cell cycle progression and apoptosis of rat liver cell line BRL-3A in vitro.Methods_MTT method was used to observe the effect of insulin on cell activity, and flow cytometry was used to detect cell apoptosis and cell cycle.qRT-PCR was used to evaluate the expression of related genes.Results_Human insulin induced the proliferation of BRL-3A cells in a dose-dependent manner ( P<0.05 or P<0.01);After 3 days treated by human insulin (500 nmol/L), the proportion of cells in G0/G1 phases re-markably decreased (P<0.05).Moreover, pro-apoptotic BAX was down-regulated (P<0.05), while cell prolif-eration-related gene CCNA2 was up-regulated (P<0.05).Conclusions_Human insulin may inhibit the apoptosis of BRL-3 A cell line and induce proliferation due to the down-regulated expression of BAX and up-regulated expres-sion of CCNA2 .

The clinical efficiency of insulin pump therapy combined with Novolin 30R in patients with diabetes mellitus complicated with pulmonary infection and analysis of detection indexes / 国际检验医学杂志

Objective To investigate the clinical efficiency of Isophane Protamine Biosynthetic Human Insulin Injection (pre‐mixed 30R)(Novolin 30R) combined with insulin pump therapy in patients with diabetes mellitus complicated with pulmonary in‐fection ,and to analyze the detection indexes .Methods From Jan .2010 to Feb .2013 ,90 cases of patients with diabetes mellitus com‐plicated with pulmonary infection were selected and divided into the observation group and control group .The control group re‐ceived Novolin 30R treatment and conventional treatment ,the observation group were given insulin pump therapy based on the treatment of control group .The clinical efficiency and detection indexes were compared and analyzed .Results The cure rate and to‐tal effective rate were 51 .11% and 82 .22% respectively in the control group ,and were 84 .44% and 100 .00% in the observation group respectively .There were significant differences of both cure rate and total effective rate between the two groups (P<0 .05) . There were statistically significant differences of hospitalization time ,the time for reaching target of glucose ,insulin dose ,recovery time of body temperature ,hospital costs ,incidence rate of complications ,serum levels of sodium ,potassium and urea nitrogen ,os‐molality and PH value between the two groups(P<0 .05) .Conclusion The combination of Novolin 30R and insulin pump therapy could provide satisfactory clinical efficiency in patients with diabetes mellitus complicated with pulmonary infection .

Effect of mixed protamine zinc recombinant human insulin lispro injection on glucose metabolism, immune function in patients with type 2 diabetes and its inflammatory mechanisms / 中国生化药物杂志

Objective To investigate the mixed protamine zinc recombinant human insulin lispro injection on glucose metabolism, immune function in patients with type 2 diabetes and its inflammatory mechanisms.Methods 125 patients enrolled were randomly divided into two groups according to the random number table: control group (n =61) and observation group (n =64).The control group were received conventional treatment, observation group were received mixed protamine zinc recombinant human insulin lispro injection on the basis of control group, with a course of three months of both groups.The fasting blood glucose (FBG), 2h post prandial blood glucose (2hPG), glycated hemoglobin (HbA1c), fasting insulin, insulin resistance index ( IRI) , CD4 +, CD8 +, CD4 +/CD8 +changes and inflammatory cytokines levels were compared before and after treatment between two groups. Results The level of FBG, 2hPG, HbA1c after treatment was respectively lower than that before treatment in both groups (P<0.05), and the above indexes of observation group after treatment was respectively lower than that of control group (P<0.05).The level of fasting insulin, IRI after treatment was respectively lower than that before treatment in both groups (P<0.05), and the above indexes of observation group after treatment was respectively lower than that of control group (P<0.05).The level of CD8 +was lower, CD4 +, CD4 +/CD8 +was higher after treatment than that before treatment in both groups, respectively (P<0.05), and the CD8 +level of observation group after treatment was lower, CD4 +, CD4 +/CD8 +was higher than that of control group, respectively (P<0.05).The level of TNF-α, IL-6, CRP after treatment was respectively lower than that before treatment in both groups (P<0.05), and the above indexes of observation group after treatment was respectively lower than that of control group (P<0.05).Conclusion The mixed protamine zinc recombinant human insulin lispro injection can significantly improve glucose metabolism in patients with type 2 diabetes, and enhance immune function, reduce inflammation, thereby reducing the incidence of cardiovascular events.