Objective To explore the effect of the improved abdominal rotation card method in insulin injection. Methods A total of 100 hospitalized diabetes patients were randomly divided into control group (n=50) and observational group (n=50) according to the random number method. In the control group, insulin was injected to the subcutaneous tissue of abdomen with traditional method annular rotating method. Insulin was injected using improved abdominal rotation card method in the observational group. Compare accuracy and mastery rate of injection site rotation between the two groups. Compare fasting blood glucose (FBG), postprandial 2H blood glucose (PBG), HbA1c, the incidence of hypoglycemia and endermic induration between the two groups after three months. Results The nurses in the observation group had higher accuracy rate of the injection site rotation compared to the control group [98.6%(690/700) vs. 38.6%(270/700),χ2=584.66, P<0.01]. Mastery rate of the injection site rotation for the patients in the observation group were significantly higher than the control group [70.0% (35/50) vs. 20.0% (10/50), χ2=25.74, P < 0.01]. The incidence of endermic induration were significantly lower in observation group compared to the control group [2.0% (1/50) vs.16.0% (8/50), χ2=5.98, P < 0.01]. The incidence of hypoglycemia were significantly lower in observation group compared to the control group [4.0%(2/50) vs. 16.0%(8/50),χ2=4.00, P<0.01]. Conclusions The new abdominal rotation method in insulin injection can be a safe and effective therapy in patients with type 2 diabetes.

The M and NP genes of H5N1 avian influenza virus (A/chicken/Hubei/489/2004) were amplified by RT-PCR from viral RNA,and cloned into pMD 18-T vector respectively.The expression plasmid containing the M gene (pHM6-m) or the NP gene (pHM6-np) was then constructed by inserting the M or NP gene into the pHM6 eukaryote expression vector; the constructed plasmid was then sequenced.32 BALB/c mice (6-week-old) were divided into four groups at random.Three groups of BALB/c mice were inoculated one time the intramuscular route with either 30 μg of plasmid pHM6-m,30 μg of plasmid pHM6-np or the mixture of plasmid pHM6-m (15 μg ) and pHM6-np(15 μg) respectively.A additional group of mice were injected with 100 μ1 PBS as controls.Two weeks later,all mice were challenged with homologous H5N1 avian influenza virus,and observed in the following 12 days.The survival rates of mice in the pHM6-m group,the pHM6-np group and mixed plasmids group were 62.5% ,25.0% and 50.0%,respectively.Results showed that effective protection could be provided by either pHM6-m or pHM6-np,but pHM6-m provided a better protective effect than pHM6-np.

Grounded theory was used to summarize and analyze influencing factors and their mechanism on availability of essential medicine.Four factors which influenced the availability of essential medicine were singled out:defective top design in the essential medicine system,interactions among its policies,deviations in the policy implementation by government agencies,and deficiency of supporting policies for the system.The availability of essential medicine in rural areas was influenced by a variety of factors.The ideas and methods of the grounded theory prove helpful for this study.In the future studies,both qualitative and quantitative study should be made to perfect this model formed by the grounded theory,to identify roadblocks and underlying causes in order to provide evidence for improving availability of essential medicine in rural areas.

Aim To observe the effects of tetrahydro-biopterin ( BH4 ) on nitric oxide ( NO ) production in the kidney of type 2 diabetic nephropathy ( DN) mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in-to DN phase were divided into 2 groups:DAHP group, subjected to intraperitoneal injection of 150 mg·kg-1 DAHP (n=8);DN group, subjected to intraperitone-al injection of same dose of normal saline containing 5% DMSO ( n = 6 ) . The age-matched db/m mice ( NS group) were subjected to intraperitoneal injection of same dose of normal saline containing 5% DMSO ( n =6 ) . Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u-rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 24h urine volume, 24h urine pro-tein, BH4 , iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser-um creatinine, urine volume, urine protein, BH4 , iN-OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over-production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.

ObjectiveTo explore the effect of bone marrow mesenchymal stem cells transplantation on the deep partial thickness derma burn injury in guinea pigs.MethodsSuspension cells of MSCs were cultured in vitro and transplanted into wounds of the female guinea pig of the deep partial thickness derma burn injuring model with the concentration of 2×106cell/ml(group A),2×107cell/ ml(group B). The rate of wound healing was observed, factor Ⅷ was detected using immunohistochemistry. Y chromosome on the wound in guinea pig was detected using PCR 15,30,50 days after MSCs had been transplanted. ResultsThe rate of wound healing of groups A and B was better than that in the control group, but there was not significantly different between groups A and B. Microvessel density of wound in groups A and B was better than those in the control group on the 15th and 30th days. There was not significantly different between the group A and B. PCR results showed that Y chromosome gene was expressed on some recoveried wound of female guinea pig. ConclusionBone marrow mesenchymal stem cells transplantation is effective in deep partial thickness derma burn injury in guinea pigs.

OBJECTIVETo observe the effect of Yuquan Pill (YP) on proinflammatory cytokines in patients with type 2 diabetes mellitus (DM2).

METHODSNinety patients with DM2 were randomly divided into the treated group and the control group by the ratio of 2:1. Both groups were treated with Glucobay on the base of dietary and exercise therapy for 3 months, with YP, given to the treated group additionally 6 g four times a day. And the changes of proinflammatory cytokines, including C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), etc. were observed.

RESULTSThe levels of serum CRP,TNF-alpha and IL-6 significantly lowered after treatment in both groups (P<0.01), and the improvement in the treated group was superior to that in the control group (P<0.01).

CONCLUSIONYP could reduce the levels of the increased proinflammatory cytokines in patients with DM2.

Acarbose ; therapeutic use ; Adult ; Aged ; C-Reactive Protein ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hypoglycemic Agents ; therapeutic use ; Interleukin-6 ; blood ; Male ; Middle Aged ; Phytotherapy ; Tumor Necrosis Factor-alpha ; blood

Chronic heart failure(CHF)is the performance of end-stage cardiovascular disease and the leading cause of death in recent years.With the rapid development of medical care,the mortality rate of heart failure is still high.This is one in the two major challenges in the cardiovascular field in the 21st century.The new drug LCZ696 is a dual inhibitors of angiotensin receptor blockers(ARB)and neprilysin(NEP),which may lead to new hope for patients with heart failure.In order to determine the efficacy and safety of LCZ696 in the treatment of heart failure,foreign countries have carried out some large-scale trials,such as PARAMOUT, PARADIGM,TITRATION and so on.The results of these studies reflected the superiority of LCZ696 compared with enalapril,valsartan and other drugs in the treatment of chronic heart failure.ARB/antiotensin converting enzyme inhibitors(ACEI)targets the angiotensin receptor to dilate blood vessels and inhibits the sympathetic nerve,but their effects on sodium withdrawal and diuresis are weak.The sacubitril in LCZ696 prevents natriuretic peptide from degrading,strengthens the natriuretic diuretic and further expansion of blood vessels.Thereby it improves water and sodium retention and cardiac function.It can play a better synergistic role combined with valsartan.

Objective To evaluate the practicability of using CRISPR/Cas9 genome editing tech-nology for inhibition of hepatitis B virus ( HBV) replication. Methods Two sgRNA targeting sites were de-signed for the S region of HBV genome. The CRISPR/Cas9 expression plasmids specific for HBV were con-structed and then transfected into a cell line expressing HBV genome(HepG2-N10). The cytotoxicity of cells transfected with different expression plasmids were detected by MTT assay. The levels of hepatitis B surface antigen ( HBsAg ) were determined by using chemiluminescent immunoassay ( CLIA ) . The expression of HBV at mRNA level was analyzed by quantitative real-time PCR ( qRT-PCR) . The qPCR was performed for the detection of extracellular and intracellular HBV DNA. The next-generation sequencing ( NGS) Illumina MiSeq Platform was used to analyze HBV genome editing. Results No significant cytotoxic effects were de-tected in HepG2-N10 cells transfected with different expression plasmids. Compared with the cells carrying pCas-Guide-GFP-Scramble, the levels of HBsAg in the supernatants of transfected cell culture harboring pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were decreased by 24. 2% (P<0. 05) and 16. 9% (P>0. 05), respectively. The levels of HBsAg in cells transfected with pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were respectively decreased by 16. 4% (P>0. 05) and 32. 1% (P>0. 05) as compared with that of pCas-Guide-GFP-Scramble transfected group. The expression of HBV at mRNA level was inhibited as indica-ted by the results of qRT-PCR. Moreover, the levels of extracellular HBV DNA were respectively suppressed by 23% (P>0. 05) and 35% (P<0. 05), and the levels of intracellular HBV DNA were respectively sup-pressed by 7. 2% (P>0. 05) and 18% (P>0. 05). Different types of insertion/deletion mutation were de-tected in HBV genome by high-throughput sequencing. Conclusion HBV-specific CRISPR/Cas9 system could inhibit the expression of HBV gene and the replication of virus. Therefore, the CRISPR/Cas9 genome editing technology might be used as a potential tool for the treatment of persistent HBV infection.

Objective To construct a bicistronic expression vector containing HPV type 6b L1 gene, to express the recombinant vector in mammalian cells, and to establish a cell strain stably expressing HPV6b L1 gene. Methods After endonuclease digestion and purification, the gene fragment of HPV6b L1 was cloned into the eukaryotic expression vector pIRES2-enhanced green fluorescent protein (EGFP). The identification of the recombinant was realized via endonuclease digestion and sequence analysis. Then, the recombinant plasmid pIRES2-HPV6bLl-EGFP was transfected into NIH3T3 (a mouse embryonic fibroblast cell line) cells. Subsequently, the expression of EGFP was observed by fluorescent inverted microscopy, and HPV6b L1 mRNA expression by reverse transcription (RT)-PCR. Results The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed, transfected into N1H3T3 cells, and selected by G418. The expression of EGFP was seen under an inverted fluorescence microscoy. RT-PCR proved the expression of HPV6b LI mRNA in transfected cells. Conclusions The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed and transfected into NIH3T3 cells. Inverted fluorescent microscopy and RT-PCR confirmed the successful expression of HPV6b L1 in NIH3T3 cells.

Objective To study the value of MSCT in the diagnosis of neonatal type (Ⅲ) congenital esophageal atresia.Methods MSCT data of 25 cases with neonatal type (Ⅲ) congenital esophageal atresia confirmed by surgery and pathology were analyzed retrospectively.The full preparation before examination and the image post-processing function were checked,and MSCT data were compared with the findings of surgery.Results By full preparation and application of M PR,SSD technology after scanning,the esophageal proximal blind end,and the distance between the two esophageal blind ends were clearly displayed,furthermore,the opening position,morphology and diameter of the distal tracheoesophageal fistula were also showed,and bronchial tree morphology and inflammation in the lung can be visible.Conclusion MSCT full preparation and application of image post processing function are potential to the clinical diagnosis of congenital esophageal atresia and tracheoesophageal fistula.