Objective To investigated the relationship between serum matrix metalloproteinase-9(MMP-9) and carotid atherosclerosis(AS) in type 2 diabetes mellitus(T2DM).Methods A total of 93 patients with T2DM were recruited to our study.The intima-media thickness(IMT) and plaques of carotid artery were measured.These patients were divided into 3 groups according to their IMT values: diabetes mellitus(DM) group(n=32),carotid artery intima thicken group(n=31) and carotid artery intima plaque group(n=30).At the same time,30 healthy individuals were selected as control.Serum level of MMP-9 were determined and analyzed.Results The serum MMP-9 in DM group was significantly higher than that in healthy controls([550.26±269.28]μg/L vs.[359.70±215.62]μg/L,t=2.23,P＜0.05).The serum MMP-9 level of intima thicken group(712.15±340.47)μg/L was significantly higher than that in healthy controls(t=4.53,P＜0.01) and DM group(t=2.40,P＜0.05).The serum MMP-9 level of plaque group([889.08±247.80]μg/L) was even more significantly higher than DM group(t=4.89,P＜0.01),IMT group(t=2.53,P＜0.05) and healthy controls(t=7.01,P＜0.01).Conclusion The severity of carotid atherosclerosis in T2DM is closely associated with the serum MMP-9 level.

Objective To study the effect of short-term inte ns ive insulin treatment on the pancreatic B-cell function in type 2 diabetic mell itus (T2DM).Methods Insulin and C-peptide serum levels were measured befor e and after intensive insulin treatment in T2DM by chemical luminescence immunoa ssay.Results The levels of insulin and C-peptide were increased at 0、30、60、120、180min in OGTT in T2DM after insulin intensive treatment.Conclusion Short-term intensive insulin treatment can partiall y improve the B-cell function of T2DM with lower function and made it rest.

Objective To study the levels of serum resistin and sICAM-1 in diffferent glucose tolerance status.Methods Fasting serum resistin and sICAM-1 concentration were measured by enzyme immunoassay in 52 cases suffering from T2DM,18 cases with glucose tolerance impaired and 30 cases with normal glucose tolerance.Meanwhile,fasting blood glucose,insulin and lipids were determined.HOMA-IR was calculated.Results Fasting serum resistin concentration (ng/mL) was higher in IGT than that in those patients with T2DM and NGT (P

Using HB55 McAb, we detected HLA-DR antigens on Daudi, H18, PBL and K562 cells by dot-ELISA. Results corresponding to situation of the antigen expressions on various cell lines were obtained: colour reaction in Daudi, H18 cells and PBL was positive in different degree and K562 sample presented negative reaction. Chromatogram peaks were printed by Dual-Wavelength Flying-Spot scanner CS-9000 (SHIMADZU). Areas and high values of peaks, meaning size and colour of spot respectively, were increased with cell concentration. Both cell concentration and volume of cell suspension did not influence the diffusion of sample. An analysis for the antigen expression on cell surface and its dynamic change can be supplied by the technique.The study on quantitative assay of MHC class Ⅱ antigens on cell surface are still going on.

Objective To study the clinicopathological features of C3 glomerulonephritis and the associations between plasma complement fragments level and clinical manifestations.Methods The clinical and pathological data of the 12 patients with C3 glomerulonephritis in our division from January 1999 to June 2010 were analyzed retrospectively.Concentrations of plasma factor B,Ba,C3,C3a,C4a and C5a were detected by commercial available sandwich ELISA kits on the day of renal biopsy.Results Ten of the 12 patients were C3 glomerulonephritis without MPGN,and the rest 2 were C3 glomerulonephritis with MPGN.All the patients presented proteinuria.Two of the 10 C3 glomerulonephritis patients without MPGN presented nephrotic range proteinuria,6 with microhematuria,1 with gross hematuria,and 2 with renal insufficiency.One of the 2 C3 glomerulonephritis patients with MPGN presented nephrotic range proteinuria,accompanied by microhematuria,hypertension and renal insufficiency.The other patient showed moderate proteinuria with normal renal function.Most of C3 glomerulonephritis patients without MPGN showed mild mesangial proliferative glomerulonephritis,and 4/10 patients had various degree cresentic formation.One C3 glomerulonephritis patient with MPGN had 47.1％ cresentic formation.The concentration of plasma C3 in C3 glomerulonephritis patients was normal,while the plasma factor B was significantly decreased,and the concentrations of plasma Ba,C3a,C4a and C5a were significantly elevated.The concentration of plasma Ba was positively correlated with the proteinuria level,while the concentrations of plasma C3a,C4a and C5a were not correlated with the levels of proteinuria or plasma creatinine.Conclusions Majority of these 12 patients were C3 glomerulonephritis without MPGN,manifests as nephritic syndrome clinically and mild mesangial proliferative glomerulonephritis histopathologically.Complement activation via alternative pathway may play an important role in the pathogenesis of C3 glomerulonephritis.

Objective To observe the change of endothelium-dependent vasodilatation function and to explore its mechanism in insulin-resistant (IR) rats induced by high fat feed. Methods (1) IR rat model was established by high fat feed for 4 weeks and IR was evaluated by glucose infusion rate (GIR) of euglycemic hyperinsulinemic clamp technique. (2) Acetylcholine (Ach)-dependent vasodilation response and nitric oxide synthase (NOS)-nitric oxide (NO)-cGMP function status were observed in isolated aorta of rats. Results (1) GIR was obviously lower in high fat feed group than that in routine feed group (P

Robot-assisted surgery is becoming a widely popular technology and now entering total knee replacement. The development of total knee replacement is introduced in this paper with the emphasis on femur positioning technologies. The experiments and error analysis of robot-assisted surgical system for total knee replacement demonstrate that femur positioning technologies have a high precision and thus deserve to be popularized theoretically and practically.

Objective To investigate the use of salvianolate injection for inpatients in our hospital; To primarily evaluate its rationality and security in clinic.Methods Medical records using the salvianolate injection from December 2013 to March 2014 were surveyed statistically and analyzed in respect of patient gender and age, department, disease diagnosis, usage and dosage, solvent types, treatment course, medicine combination, occurence of ADR, etc.Results Totally 600 cases of inpatients receiving salvianolate injection were collected, about 69.33% consistent with the indication, 67.00% with a correct single dosage, and 52.50% with a correct treatment course. In the clinical application of salvianolate injection, a high rate of unreasonable usage was found in indication, single dosage, treatment course and medicine combination.ConclusionBecause the problems still existed in the use of salvianolate injection, it should be used strictly according to the medicine instructions and General Principles for Clinical Application of Traditional Chinese Medicine Injection to ensure clinical medication safety.

OBJECTIVETo determine the regulatory effects of the circadian clock gene Per1 on cell cycle-related genes and its influence on the proliferation, apoptosis, cycle, and tumorigenicity in vivo of human oral squamous cell carcinoma SCC15 cells.

METHODSThree groups of the short hairpin RNA (shRNA) of lentivirus recombinant plasmids were designed against the RNA of Per1 and then transfected to the SCC15 cells. The optimum interference group was screened through Western blot and quantitative real-time PCR (qRT-PCR) and assigned as the experimental group. The transfected lentivirus plasmid without an interference effect on any gene was set as the control group (Control-shRNA). Untreated SCC15 cells were set as the blank group. The mRNA expressions of cell cycle-related genes, namely, Per1, p53, Cyclin D1, Cyclin E, Cyclin A2, Cyclin B1, CDK1, CDK2, CDK4, CDK6, p16, p21, Wee1, cdc25, E2F, and Rbl1 in each group were detected through qRT-PCR. The cell proliferation, apoptosis, and cell cycle distribution in each group were evaluated through flow cytometry. The cells of the experimental group and the blank group were subcutaneously inoculated in nude mice to observe tumorigenesis.

RESULTSThree groups of Per1-shRNA lentivirus plasmids were constructed successfully. Among the groups, the Per1-shRNA- I group exhibited the highest interference effect, as indicated by qRT-PCR and Western blot analysis. As such, this group was set as the experimental group. The mRNA expression levels of CyclinD1, CyclinE, CyclinB1, CDK1, and Wee1 gene in the Per1-shRNA-I group were significantly higher than those in the Control-shRNA group and the SCC15 group (P < 0.05). By contrast, the mRNA expression levels of p53, Cyclin A2, p16, p21, and cdc25 in the Per1-shRNA-I group were significantly lower than those in the Control-shRNA group and the SCC15 group (P < 0.05). The mRNA expression levels of each gene between the Control-sLRNA group and the SCC15 group did not significantly differ (P > 0.05). The mRNA expression levels of CDK2, CDK4, CDK6, E2F, and Rb1 did not significantly differed in the three groups (P > 0.05). The proliferation index of the Perl-shRNA-I group was significantly higher than those of the Control-shRNA group and the SCC15 group (P < 0.05). The apoptosis index of the Per1-shRNA-I group was significantly lower than those of the Control-shRNA group and the SCC15 group (P < 0.05). The number of S-phase cells in the Per1-shRNA-I group was significantly lower than those of S-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). The number of G2/M-phase cells in the Per1-shRNA-I group was significantly higher than those of G2/M-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). Conversely, the proliferation index, apoptotic index, and cell cycle distribution of the cells in the Control-shRNA group did not significantly differ from those of the SCC15 group (P > 0.05). The tumorigenic ability in vivo was significantly enhanced in the Per1-shRNA-I group (P < 0.05).

CONCLUSIONPer1 is an important tumor suppressor gene. Per1 can regulate a large number of downstream cell cycle-related genes. The alteration of its expression can affect cell cycle progression, proliferation, apoptosis imbalance, and tumorigenic ability in vivo. Further studies on Per1 may elucidate cancer development and provide novel effective molecular targets for cancer treatment.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Circadian Clocks ; genetics ; Cyclin D1 ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Period Circadian Proteins ; genetics ; Plasmids ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Transfection

To construct an HSV-1 vector vaccine carrying HIV-1 antigens, HIV-1 gp160, gag, protease and the expression elements were chained together, and then inserted into the internal inverted repeat sequence region of HSV-1 by bacterial artificial chromosome technology. Firstly, HIV-1 gp160 (including type B and C), gag and protease genes were cloned into pcDNA3 in series to generate the pcDNA/gBgp and pcDNA/gCgp, then the recombinant plasmids were transfected into 293FT cells, and HIV-1 antigen was detected from transfected cells by Western blotting. Then the expression cassettes from pcDNA/gBgp and pcDNA/gCgp, comprising HIV-1 antigen genes and expression elements, were cloned into pKO5/BN to generate the shuttle plasmids pKO5/BN/gBgp and pKO5/BN/gCgp. The shuttle plasmids were electroporated into E. coli cells that harbor an HSV-BAC, the recombinant bacteria were screened, and the recombinant DNA was extracted and transfected into Vero cells. The recombinant virus was purified through picking plaques, the virus' DNAs were identified by Southern blotting; HIV-1 antigen was detected from the recombinant HSV-1 infected cells by Western blotting, and the virus' replication competent was analyzed. As the results, gp160 and gag proteins were detected from 293FT cells transfected with pcDNA/gBgp and pcDNA/gCgp by Western blotting. The recombinant bacteria were generated from the E. coli electroporated with pKO5/BN/gBgp or pKO5/BN/gCgp. The recombinant HSV was purified from the Vero cells transfected with the recombinant DNA, the unique DNA fragment was detected from the genome of recombination HSV by Southern blotting; gp120 and gp41 were detected from the infected cells by Western blotting, and the recombinant HSV retained replication competent in mammalian cells. The results indicate that the recombinant HSV carrying HIV-1 gp160, gag and protease genes was generated, the virus retains replication competent in mammalian cells, and could be used as a replicated viral vector vaccine.
Animals ; Cercopithecus aethiops ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; genetics ; DNA, Viral ; genetics ; Escherichia coli ; HIV Antigens ; genetics ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Protease ; genetics ; immunology ; Herpes Simplex Virus Vaccines ; immunology ; Herpesvirus 1, Human ; physiology ; Plasmids ; Transfection ; Vero Cells ; Virus Replication ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; immunology