Objective To analyze the effects of alprostadil combined with benazepril in the treatment of diabetes and the effect on blood glucose fluctuation.MethodsEighty cases with diabetes were randomly divided into the control group and the observation group with 40 cases in each group.The control group was treated with benazepril while the observation group was additionally treated with alprostadil.The treatment effects, the changes of blood glucose, blood lipids and renal function were compared between the two groups, and the incidence of adverse reactions was recorded.ResultsThe total effective rate in the observation group was higher than that in the control group (P<0.05).After treatment, the fasting plasma glucose (FPG), postprandial 2h plasma glucose (2hPG) and glycosylated hemoglobin (HbA1c) were reduced to the normal level but the volatility differences of FPG and 2hPG in the observation group were lower than those in the control group (P<0.05).After 4 weeks of treatment, the total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) decreased while high-density lipoprotein cholesterol (HDL-C) increased (P<0.05) but the differences between the two groups showed no statistical significance.After treatment, the improvement of renal function indexes in the observation group was better than that in the control group (P<0.05), and the incidence of adverse reactions showed no significant difference between the two groups.ConclusionThe application of alprostadil combined with benazepril in the treatment of diabetes can reduce blood glucose fluctuation, improve renal function, and reduce the risk of diabetic nephropathy.

BACKGROUND:β-catenin is the most critical signaling molecule in the Wnt/β-catenin signaling pathway, which is involved in the regulation of cellproliferation, differentiation and tissue self-healing balance.
OBJECTIVE:To construct a stableβ-catenin over-expression lentivirus-mediated vector and to transfect mesenchymal stem cells line for investigating its effects on proliferation and migration of mesenchymal stem cells.
METHODS:Over-expression vector, PLV-EF1A-catenin-RFP, was constructed and transfected the 293T cellto infect mesenchymal stem cells, and positive cells were selected with puromycin. The up-regulated efficiency of targetingβ-catenin gene at mRNA level was detected by real-time quantitative PCR, the effect on proliferation of mesenchmal stem cellwas assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve, and the migration ability was detected by Transwel motility assay.
RESULTS AND CONCLUSION:The lentiviral vector targetingβ-catenin gene was constructed successful y, and a stable mesenchymal stem cellline that up-regulatedβ-catenin was established. Real-time quantitative PCR results showed that the expression ofβ-catenin gene was efficiently up-regulated by infecting PLV-EF1A-catenin-RFP (P<0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and growth curve showed that celldoubling time was shortened after infected with pLV-EF1A-catenin-RFP (P<0.05), indicating that the over-expression of theβ-catenin gene successful y increased the proliferative capability of mesenchymal stem cells. The Transwel assay also showed similar increasing results on the migration ability (P<0.01). The lenvivirus-mediated over-expression of theβ-catenin gene can be used to increase the proliferation and migration abilities of the mesenchymal stem cells.

Objective To construct a lentiviral vector over-expressing β-catenin gene by multisite Gateway technology and confirm its effect.Methods By using multisite Gateway clone technique,the entry clone of pDown-Ctnnb1 was constructed using BP recombination reaction.Then,LR recombination reaction was performed among pUp-EF1A,pDown-Ctnnb1,pTail-IRES/DsRed-Express2 and pLV.Des3d.P/puro to generate an expression clone of pLV.EX3d.P/puro-EF1A＞Ctnnb1 ＞IRES/DsRed-Express2.In each step,PCR and sequencing analysis were used to verify the constructions.When it was verified that plasmids were transfected into 293T cells,PT-PCR was performed to determine the mRNA level of β-catenin gene.Results Both PCR and sequencing analysis revealed that β-catenin over-expression gene was inserted into the target site and the insertion sequence was perfectly corrected.The RT-PCR results showed that the expression of β-catenin gene was significantly upregulated.Conclusion The lenvivirus-mediate β-catenin over-expression gene was successfully constructed..

BACKGROUND:Previous studies have verified that mesenchymal stem cells could be transplanted into inflammatory bowel mucosa to repair inflammatory bowel tissue. OBJECTIVE:To observe the differential gene expression in large intestine before and after mesenchymal stem celltransplantation in repair of inflammatory bowel tissue of rats using microarray technology, and to primarily discover the main genes during mesenchymal stem celltransplantation, differentiation, and reparation in inflammatory colorectal tissue region. METHODS:Healthy Sprague-Dawley rats were randomly divided into two groups. Experimental rat models of inflammatory bowel disease were established using trinitrobenzene sulfonic acid via enema. At 24 hours after model establishment, green fluorescent protein-labeled mesenchymal stem cells were infused via the caudal vein. The control group was treated with physiological saline by enema, instead of trinitrobenzene sulfonic acid. At 28 days, large intestine was obtained from the experimental group and control group. Differential y expressed genes were screened in the experimental and control groups using microarray technique. RESULTS AND CONCLUSION:The microarray analysis results showed that there were 388 differential genes in the control and experimental groups (P<0.05, FC>2), in which 191 were up-expressed, and 197 were down-expressed. Al of these genes were mainly involved in inflammatory reaction, immune reaction and celldifferentiation. In the top 10 up-regulation and down-regulation differential genes (total y 20 genes), 3 genes were involved in inflammation, 3 genes were involved in immune reaction, and 2 genes were related to stem celldifferentiation. In the 388 genes, 33 were related to signaling pathways (P<0.05), 6 related to inflammation, 8 related to immunity, and 5 related to stem celldifferentiation. Results suggested that the main genes involved in mesenchymal stem cells in repair of inflammatory bowel tissue were primarily screened using gene expression microarray technique.

Objective To explore the factors influencing the recovery of ability in the activities of daily living (ADL) after intracerebral hemorrhage. Methods A total of 108 patients with intracerebral hemorrhage admitted for rehabilitation to the rehabilitation medicine department of Huashan Hospital between January 2007 and June 2011 were studied.Twelve items of clinical data were collected with regard to the patients' medical history,physical status,modified Barthel index (MBI) score and Brunnstrom stage at admission.Functional status was classified according to the MBI scores and Brunnstrom stages assessed at admission and before discharge.Linear regression analysis was used to relate the variables. Results After rehabilitation,the MBI scores and Brunnstrom stages had improved relative to the scores at admission.Factors influencing the MBI improvements included the intervention timing of rehabilitation and the course of therapy employed. Conclusions It is very important to comprehend the factors influencing the recovery of ADL ability after cerebral hemorrhage in order to design effective rehabilitation strategies,better predict functional outcomes and improve patients' ADL ability effectively.

Objective To analyze the differential expression of Stathmin in human cells infected with human-tropic porcine endogenous retrovirus(PERV)and to explore the potential molecular effect of human-tropic PERV on human cells.Methods HEK293 cells were infected with the human-tropic PERV infectious molecular clone.PCR,real-time RT-PCR and immunofluorescence analysis were applied to confirm that HEK293 cells were infected.Then real-time RT-PCR and Western blot were carried out to analyze the differential expression of Stathmin at the mRNA level and protein level,respectively.Results HEK293 cells were infected by human-tropic PERV.Real-time RT-PCR and Western blot analysis showed that Stathmin was up-regulated in HEK293 cells infected with PERV compared with the control cells.Conclusion Stathmin was up-regulated in HEK293 cells infected with human-tropic PERV.These studies will be helpful for revealing the interaction of PERV and human cells,and for understanding the molecular effect of humantropic PERV on human cells.In addition,it suggested that PERV infection may infect cell growth and physiological functions,even be pathogenic.These will help to clarify the biologic characteristics of PERV and evaluate the safety of PERV in pig to human xenotransplantation.

Objective To explore the potential correlation between fluoride levels of urine and influencing factors in water-high-fluoride areas.Methods In 2010,based on plain area,mountainous area and mixed area (altitudes were 898,2 200,1 460 m,respectively),3 villages of water-high-fluoride areas were selected by purpose sampling;pupils' family members were selected as survey population by cluster sampling.Questionnaire was conducted to collect general information;fluoride contents in urine and drinking water were measured using ionselective electrode.A linear multiple regression was used to examine which factors affected urinary fluoride.Results Totally there were 968 people distributed in plain area (444),mixed area (368) and mountainous area (156),medians of urinary fluoride level were 0.71,1.59 and 1.67 mg/L,respectively,the difference was significant (F =203.90,P ＜0.01);medians of water fluoride level in the three different habitats were 0.50,1.00 and 3.50 mg/L,respectively,the difference was significant (F =331.98,P ＜ 0.01).Age,gender,fluoride contents in drinking waters and habitat explained 33.1％ of urinary fluoride variation.Male had higher urine fluoride than female,older age and higher level fluoride in drinking water contributed to higher fluoride level in urine,higher altitude contributed to higher urinary fluoride.Conclusion Once fluoride content in urine is used to estimate fluoride exposure level among people in high fluoride area,gender and age must be taken in consideration.

Objective To investigate the expression of macrophage migration inhibition factor (MIF) and cell cycle regulating factor Cyclin D1 in hepatocellular carcinoma tissue and the interaction between MIF and Cyclin D1 in hepatocellular carcinoma cell cycle controlling. Methods Using quantitative real-time PCR and Western blotting to detect mRNA and protein expression of MIF and Cyelin DI in HCC tissues and tumor adjacent tissues. Specific small interfering RNA(siRNA) targeting MIF gene was transfccted at doses of 50 nmol/L and 100 nmoL/L into HCC cell lines of PLC and HepG2 with lipofeetamine 2000 methods to knockdown the expression of M1F gene and to investigare the the interaction between M1F and Cyclin D1. Results MIF and Cyclin D1 protein and mRNA were overexpressed in HCC tumor tissues. The relative expression of MIF,Cyclin D1 protein and mRNA were 0.825±0.13,0.843± 0.104 and 7.31±1.85 folds、4.27±1.05 folds, compared with the tumor adjacent tissues (FMIF= 15.5, P<0.01;FCyclin D1=87.5,P <0.01). In MIF siRNA treated PLC and HepG2 cells, MIF mRNA down regulation 71.2%±7.2%, 87.4%±2.9% ,74.3%±8.9% and 88.4%±4.6% respectively (FPLC = 315.5 ,P < 0.01 ; FHepG2= 201.2 P < 0.01). While MIF protein expression were significandy reduced to 0.33±0.03,0.11±0.02, 0.81±0.08 and 0.36±0.02 in a dose-dependent manner (FPLC= 43.9, P <0.01 ;FHepG2 = 133.4 P <0.01). Cyclin D1 mRNA was significantly down-regnlated in MIF siRNA treated PLC and HepG2 cell lines when compared with control group(P <0.01). In 50 nmol/L and 100 nmol/L groups, Cyclin DI mRNA levels were respectively decreased by 68.2%±3% and 78.1%±1.4% in PLC cell, 65.8%±4.7% and 77.3%±2.6% in HepG2 cell (FPLC= 1569, P < 0.01 ; FHepG2= 480.4, P <0.01). Compared with control groups, Cyclin D1 protein levels significantly reduced to 0.28±0.06、0.15±0.03 and 0.44 ±0.04、0.13±0.02 in the PLC and HepG2 after M IF siRNA treatment(FPLC= 35.5, P < 0.01 ; FHepG2 = 114.7, P < 0.01). Conclusions MIF and Cyclin D1 mRNA and protein were overexpressed in HCC tumor tissues and participated in tumor cell cycle regulation. MIF may up-regnlate the expression of Cyclin DI via ERK signalling and precipitate in carcinogenesis of hepatocellular carcinoma.

Objective To assess the clinical significance of endoscopic ultrasonography (EUS) in the diagnosis and preoperative staging of gastric cancer. Methods EUS was carried out in 22 patients inclu-ding 17 gastric cancer patients and 5 patients in suspicion. Helical CT scanning was performed in all of the patients and fine needle aspiration biopsies ( FNAB) were administrated to 5 suspicious patients. Compared the results of operation and pathology with those of tumor staging by estimating the depth of tumor invasion ( T) , local lymph node metastasis ( N) and metastasis to neighboring or remote organs ( M) in order to esti-mate the accuracy of diagnosis and TNM staging. The sensitivity and specificity of tumor-node-metastasis staging of gastric cancer by EUS were compared with those of the spiral CT according to the final histopatho-logical results. Results In 5 suspicious patients specimens were successfully obtained by FNAB under the guide of EUS with the pathological diagnosis of adenocarcinoma in 4 cases and signet ring cell carcinoma, 1 case. All patients underwent radical gastrectomy except one in T1N0M0, staging was treated by endoscopic mucosal resection (EMR). The sensitivity and specificity of EUS in T, N, and M stage were 84.9% and 74. 2% , 92. 1% and 77. 1% , 63. 4% and 87. 5% respectively; whereas those of CT in T, N, and M stage were 27. 3% and 75% , 31.5% and 100% , 50% and 100% respectively. The sensitivity of EUS in T and N staging were higher than those of CT with significant statistical difference (P

OBJECTIVE: To study the effects of myo-inositol and luteolin on human lung cancer A549 cells and explore the possible mechanisms. METHODS: A549 cells were treated with different concentrations of myo-inositol and luteolin, either alone or in combination, and the cell viability was examined using MTT assay. A549 cells and human bronchial epithelial Beas-2B cells were treated for 48 h with 10 mmol/L myo-inositol and 20 μmol/L luteolin, alone or in combination, and the cell proliferation was detected using MTT assay; the colony formation and migration of the cells were examined with colony formation assay and wound healing assay, respectively. The protein expression levels in A549 cells were detected using Western blotting. RESULTS: Both myo-inositol and luteolin could dose-dependently inhibit the viability of A549 cells. Treatments with 10 mmol/L myo-inositol, 20 μmol/L luteolin, and both for 48 h caused significant reduction in the cell viability (92%, 83% and 70% of the control level, respectively) and colony number (79%, 73% and 43%, respectively), and significantly lowered the wound closure rate (24.61%, 13.08% and 8.65%, respectively, as compared with 29.99% in the control group). Similar treatments with myoinositol and luteolin alone or in combination produced no significant inhibitory effect on the growth, colony formation or migration of Beas-2B cells. The expressions of p-PDK1 and p-Akt in myo-inositol-treated A549 cells and the expression of pPDK1 in luteolin-treated cells were significantly decreased ( < 0.05), and the decrements were more obvious in the combined treatment group ( < 0.05). CONCLUSIONS Luteolin combined with myo-inositol can selectively inhibit the proliferation and migration of A549 cells, and these effects are probably mediated, at least in part, by suppressing the activation of PDK1 and Akt.
A549 Cells ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Humans ; Inositol ; administration & dosage ; therapeutic use ; Lung Neoplasms ; drug therapy ; metabolism ; Luteolin ; administration & dosage ; therapeutic use ; Protein-Serine-Threonine Kinases ; drug effects ; metabolism ; Proto-Oncogene Proteins c-akt ; drug effects ; metabolism ; Vitamin B Complex