Objective To explore the clinical value of the insulin pump(CSⅡ) and multiple subcutaneous insulin intensive treat-ment(MDI) for patients with type 2 diabetes and stroke .Methods A total of 80 patients with type 2 diabetes and stroke in Weifang Community Health Service Center from April 2011 to May 2014 were selected as subjects and divided into experimental group and control group according to random number table method ,with 40 cases in each group .Patients in the experimental group were trea-ted with CSⅡ ,patients in the control group were treated with MDI .Glucose level ,therapeutic time ,the number of low blood sugar , insulin dosage ,National Institute of Health Stroke Scale(NIHSS) score ,insulin secretion index(IS) ,insulin resistance index(IR) , hospital costs were compared before and after treatment in the two groups .Results The glucose levels ,therapeutic time ,the num-ber of low blood sugar ,insulin dosage ,NIHSS score ,IS ,IR in the two groups before and after breakfast ,before and after lunch ,be-fore dinner ,after dinner 2 hours before treatment had no significant differences(P>0 .05) ,the glucose levels ,therapeutic time ,the number of low blood sugar ,insulin dosage ,NIHSS score ,IS ,IR after treatment in both groups were significantly decreased at these time points ,but those indicators in the experimental group decreased more significantly ,and had significant differences with those of the control group(P<0 .05) .The cost of hospitalization of patients in the experimental group was significant lower than that in the control group(P<0 .05) .Conclusion CSⅡ treatment for type 2 diabetes and stroke patients ,not only control glucose level well ,re-duce the number of low blood glucose and insulin dosage ,significantly improved IS and IR ,reduce hospital costs ,and also could sig-nificantly improve the neurological disorders of consciousness ,which is an ideal treatment method compared with MDI .

This study aimed to construct full-length cDNA clones of the Japanese encephalitis virus (JEV). SA14-14-2 strain and discuss the feasibility of constructing chimeric viruses for exogenous gene expression based on the JEV genetic skeleton. Long-fragment RT-PCR techniques were applied to amplify JEV cD-NAs, and two amplified fragments with corresponding restriction endonuclease sites at both ends were cloned into the pACYC184 vector sequentially. Using standard molecular techniques, the enhanced green fluorescent protein (EGFP) gene was inserted into the 3' non-coding region of JEV as a reporter gene. After in vitro transcription and transfection procedures, wild-type JEV and chimeric JEV that expressed the EGFP as the reporter gene were successfully rescued. The recovered viruses were characterized by RT-PCR, plaque assays, and direct fluorescence microscopy. After six serial passage generations, the stability of the recovered viruses were studied in terms of virus growth characteristics and structural gene expression. The results showed that cDNA clones of rJEV and rJEV-EGFP were successfully constructed and rescued in BHK-21 cells after in vitro transcription and transfection. Each generation of the recovered viruses was stable and the chimeric virus rJEV-EGFP could stably express EGFP. The findings of this study indicate that both rJEV and rJEV-EGFP could be constructed and rescued in BHK-21 cells, and the JEV SA14-14-2 strain could be obtained as a viral vector to express foreign genes.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Encephalitis Virus, Japanese ; genetics ; metabolism ; Encephalitis, Japanese ; virology ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Vaccines, Attenuated ; genetics ; metabolism ; Viral Vaccines ; genetics ; metabolism

Objective To evaluate the value of high frequency ultrasound in sciatic nerve of rat with diabetic neuropathy.Methods Fifty male Sprague-Dawley rats were divided into 4 groups:Diabetic animals were induced by intravenous injection of streptozotocin after high fat feed for 1 month,after which rats were grouped as 3-month (n =12) and 5-month (n =20) diabetes.Control group were received the streptozotocin vehicle and were classified as 3-month group (n =8) and 5-month group (n =10).Ultrasounic study of cross sectional area (CSA) and pathologic study including number and area of nerve fiber were performed at 3 and 5 months according to the disigned.Results The CSA,number and area of nerve fiber in 5 month diabetes were significantly reduced than 5-month control group (t =5.121,P ＜ 0.05; t =7.113,P ＜0.05; t =6.328,P ＜0.05).The CSA were statistically with the number and area of nerve fiber in 5 month diabetic group (r =0.732,P ＜0.05 ; r =0.715,P ＜0.05).However,the CSA had no significant correlation with the number and area of nerve fiber between 3-month diabetes and control group (P ＞0.05).Conclusions The CSA of sciatic nerve can evaluate the diabetic neuropathy of rat,which might be regarded as a potential index in diagnose of diabetic neuropathy.

Objective To study the clinical therapeutic effectiveness and the mechanism of Shouti Oral Liquid on simple obesity children with hyperinsulinism. Methods Fourty - three patients were divided randomly into the treatment group (TG) and the control group (CG). TG took Shouti Oral Liquid and CG took metformin. The course of therapy lasted 6 months. The plasma lipid, body fats, insulin resistance index (IR) and some hormones related to obesity before and after therapy were measured and the changes were analyzed. Results The weight, F% , body mass index (BMI), leptin(LP), tumor necrosing factor (TNF - ?), FINS, IR all declined markedly than pre - therapy in both groups. The significant effect, normal effect and total efficiency were 22.7 % , 59.1 %, 81.8 % respectively in TG and9.5%,52.4%,61.9% in CG. The effect in TG was better than that in CG. A short course with Shouti Oral Liquid therapy did not have obvious side effect Conclusions There are lipid metabolism disorder, insulin resistance and high lever LP and TNF - ? obesity children with hyperinsulinism. The total effect with Shouti Oral Liquid is better than that with medicine in the control group and the mechanism was regulating lipid metabolism, decreasing LP and TNF-? and ameliorating insulin resistance.

Objective To investigate protective activity against Aβ25-35-induced cytotoxicity in PC12 cells of different extracts and ursolic acid, which were isolated from pyrola decorata. Methods Aβ25-35-induced cytotoxicity in PC12 cells was established as the model in vitro. The cultured PC12 cells were divided into blank control group, DMSO control group, model control group, different extract groups of pyrola decorate and ursolic acid(UA) group. The different extract groups included ether extract (PE), acetidin extract (AE), n-butanol extract (BE), the water extract (WE), 50% ethanol extract (HEE). MTT assay was used to test the optimum concentration, and the number of viable cells in culture medium was measured by ELISA at 490 nm wavelength. Results The cell viabilities in different extracts groups(PE, AE, BE, WE, HEE) were respectively 89.3%, 77.2%, 79. 2%, 75. 1% ,74. 0% at the concentration of 5. 0 mg ? mL-1 . Moreover, ursolic acid showed the best neuroprotective activity (88.9%) at the concentration of 500 μg?L-1 . Compared with model control group, the survival rate of each group was remarkably increased, and the protective activities of PE and UA were more significant among them. Conclusion Different polar extracts of pyrola decorata and isolated ursolic acid have neuroprotective effects on Aβ25-35-induced cytotoxicity in PC12 cells in certain degrees.

Objective To evaluate the morphological changes of sural nerve in patients with type 2 diabetes mellitus by hyperfrequency ultrasound.Methods Fifty-six sural nerves of symptomatic group,64 sural nerves of asymptomatic group,and 60 sural nerves of control group were identified by 22 MHz Ultrasound.The thickness/width (T/W) ratio,cross-sectional areas (CSA) and the maximum thickness of neuronal fascicles (MT) of the sural nerve were calculated in transverse sonograms.Results ①Ultrasound can clearly show these structures of sural nerve such as epineurium,perineurium and nerve bundles and so on.②In symptomatic group,asymptomatic group and control group,T/W ratio and MT were gradually increased,and the differences among the three groups were statistically significant (P＜0.05).Whereas,there were no statistically significant differences in CSA among the three groups (P=0.257).Conclusions22 MHz ultrasound may be a valuable tool in evaluating the diabetic cutaneous nerve neuropathy

Anti-cancer drug bleomycin (BLM) can cause acute lung injury (ALI) which often results in pulmonary fibrosis due to a failure of resolving acute inflammatory response. The aim of this study is to investigate whether toll-like receptor (TLR) 2 mediates BLM-induced ALI, inflammation and fibrosis. BLM-induced dendritic cells (DCs) maturation was analyzed by flow cytometry and cytokine secretion was detected by the ELISA method. The expression and activity of p38 and ERK MAPK were determined with Western blotting. The roles of TLR2 in ALI, inflammation and fibrosis were investigated in C57BL/6 mice administered intratracheally with BLM. The results demonstrated that BLM-administered mice had higher expression of TLR2 (P<0.001) and its signaling molecules. Blocking TLR2 significantly inhibited the maturation of DCs and reversed BLM-stimulated secretion of cytokines in DCs, such as IL-6 (P<0.001), IL-17 (P<0.05) and IL-23 (P<0.05). TLR2 inhibition attenuated BLM-induced increase of inflammatory cells in bronchoalveolar lavage fluid (BALF), and reversed the immunosuppressive microenvironment by enhancing TH1 response (P<0.05) and inhibiting TH2 (P<0.001), Treg (P<0.01) and TH17 (P<0.01) responses. Importantly, blocking TLR2 in vivo significantly protected BLM-administered mice from pulmonary injury, inflammation and fibrosis and subsequently increased BLM-induced animal survival (from 50% to 92%). Therefore, TLR2 is a novel potential target for ALI and pulmonary fibrosis.

The nuclear transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) plays a crucial role in maintaining cellular redox homeostasis. The aberrant NRF2 signaling confers enhanced antioxidant capacity, which is linked to tumor progression and therapeutic resistance. The current study investigates the biological effects and molecular mechanism of tribbles homolog 3 (TRIB3), a stress-induced protein, in regulating cell survival and apoptosis in lung cancer. This study first performed the RNA sequencing data analysis with 576 lung adenocarcinoma patients from the cancer genome atlas (TCGA) database. The NRF2- antioxidant response element (ARE) signature was enriched in patients with high TRIB3 expression. Dual-luciferase reporter assay and real-time quantitative polymerase chain reaction (PCR) were used to confirm the effect of TRIB3 on the kelch-like ECH-associated protein-1 (KEAP1)-NRF2 pathway. Abrogation of TRIB3 impaired NRF2 transcriptional activity and reduced the expression of its target genes. Moreover, TRIB3 enhanced NRF2 stability via blocking KEAP1-NRF2 interaction. TRIB3-depletion promoted reactive oxygen species (ROS) production, restrained cell proliferation, and enhanced carboplatin-induced apoptosis. In addition, NRF2 overexpression recovered the tumor inhibition effect of TRIB3-depletion. Consistently, TRIB3 failed to modulate apoptosis in NRF2 depletion cells. In summary, this study shows that TRIB3 inhibits the KEAP1-NRF2 interaction and upregulates the transcriptional activity of NRF2, thereby promoting lung cancer cell proliferation and reducing the sensitivity to chemotherapy. Targeting the TRIB3-NRF2 signal axis may become a new strategy for ROS homeostasis and lung cancer treatment.

OBJECTIVE To investigate how MLKL functions on the membrane and explore its electrophysiological characters and structure. METHODS The full-length human MLKL were expressed in SF21 cells and purified using glutathione-sepharose affinity chromatography. The currents of purified MLKL proteins were recorded in avoltage-clamp mode using a Warner BC-535 bilayer clamp amplifier. The currents were digitized using pCLAMP 10.2 software. HEK293 cells were cultured and transfected with MLKL plasmid. Cell viability was examined using the CellTiter- Glo Luminescent Cell Viability Assay kit. RESULT MLKL forms cation channels that are permeable preferentially to Mg2+ rather than Ca2+ in the presence of Na+ and K+. Moreover,each MLKL monomer contains five transmembrane helices:H1, H2, H3 , H5 and H6 of the N-terminal domain which is sufficient to form channels. Finally, MLKL-induced membrane depolarization and cell death exhibit a positive correlation to its channel activity.

Objective To explore the value of 11C-choline PET/CT in patients with hepatic spaceoccupying lesions that have an indeterminate diagnosis by 18F-fluorodeoxyglucose (FDG) PET/CT. Methods A total of 25 liver masses in 20 patients with an indeterminate diagnosis based on 18F-FDG PET/CT were enrolled. Regional 11C-choline PET/CT scan was performed in all of the patients. Lesions with intense 11C-choline uptake were considered as positive. The semiquantitative maximum standardized uptake value(SUVmax) was measured and the tumor-to-liver (T/L) radioactivity ratio was calculated. The Mann-Whitney test,Kruskal-Wallis test and crosstabs x2-test were performed by using SPSS version 11.5. Results Of the 25 lesions,21 were proven to be hepatocellular carcinomas (HCC),3 hemangiomas,and 1 parasitic granuloma. The sensitivity of 11C-choline PET/CT for the detection of HCC was 66.7% (14/21). 11C-choline PET/CT had a higher sensitivity for well differentiated HCC than moderately and poorly differentiated HCC on a patient basis (8/9 vs 2/5,respectively). There were significant differences of 11C-choline T/L ratios between the HCC positive group,HCC negative group and benign lesion group ( 1.70 ± 0.35,0.86 ± 0.15,and 0.36 ± 0.18,x2 = 19.00,P ＜0.01 ). The lesion size and alphafetoprotein (AFP) level between the HCC positive and negative groups had no significant difference respectively ( Mann Whitney U = 39.00,P ＞0.05,and U=16.00,P＞0.05,respectively). Conclusions 11C-choline is complementary to 18F-FDG PET/CT for the detection of HCC,especially for well differentiated HCC.