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Objective:To observe the therapeutic effects of human umbilical cord mesenchymal stem cells (HUC-MSCs) on insulin resistance, and to investigate the molecular mechanisms in T2DM rats.Methods:The T2DM rats were induced by a high fat and high glucose diet for 10 weeks combined with low-dose streptozocin. Four weeks after infusion of HUC-MSCs via tail vein of the rats, fasting blood glucose, triglycerides, cholesterol were measured. Intraperitoneal glucose tolerance test, intraperitoneal insulin tolerance test and hyperinsulinemic-euglycaemic clamp test were performed to evaluate the islet function and insulin resistance level of rats. The protein expression levels of lipid metabolism signal pathway adenine monophosphate activated protein kinase (AMPK) and acetyl CoA carboxylase (ACC) in liver tissue were detected by western blot.Results:Compared with the T2DM group, HUC-MSCs treatment can significantly reduce fasting blood glucose, triglycerides, total cholesterol levels ( P<0.01), and the values of area under the curve of glucose tolerance and insulin tolerance ( P<0.05) in the T2DM+ HUC-MSCs group. Hyperinsulinemic-euglycaemic clamp test found that compared with the T2DM group, after HUC-MSCs treatment, the glucose infusion rate level was significantly higher in the T2DM+ HUC-MSCs group( P<0.01); Western blot showed that compared with the T2DM group, the ratio of p-AMPK to AMPK and p-ACC to ACC in liver tissues of T2DM+ HUC-MSCs group were significantly increased( P<0.01). Conclusion:Human umbilical cord mesenchymal stem cells treatment may improve lipid metabolism and insulin resistance by activating AMPK/ACC signaling pathways in type 2 diabetic rats.
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Objective@#To obtain the serotype diversity and antimicrobial resistance of Salmonella isolates recovered from retail chicken carcasses for sale in six regions of China.@*Methods@#From August 2010 to March 2012, each month 20 retail chicken carcasses including freshly slaughtered, chilled and frozen samples were collected from supermarkets and farmer's markets in 7 monitoring sites in Beijing, Jilin province, Inner Mongolia Autonomous, Shanxi province, Jiangsu province and Guangdong province, respectively. Samples were routinely collected for 12 months for each site. 1 680 chicken carcasses were collected in total and 2 629 Salmonella strains were isolated by PCR and biochemical method. Luminex xMAP method and classical slide agglutination method were carried out to determine isolates' serotypes. Minimal inhibitory concentrations (MICs) of 10 classes of antimicrobials including 14 agents were determined using broth micro-dilution method. Mocular methods were used to determine antimicrobial resistance genes of CIP-CTX-CT co-resistant isolates.@*Results@#In all, 2 629 Salmonella isolates, there were 17 seorgroups and 58 serotypes, B and D1 were the dominant serogroups with rates of 34.7% (n=913) and 31.0% (n=815), Enteritidis (30.8%, n=810), Indiana (17.6%, n=463), Infantis (10.6%, n=278) were the top three serovars. We found 224 CIP-CTX co-resistant S. Indiana containing 3 colistin resistant strains, one of them carrying mcr-1 gene and being ESBLs positive, which demonstrated a nine multi drug resistance against 11 antimicrobials tested.@*Conclusion@#These data began to describe the complicated serovar diversity and heavy antimicrobial resistance of Salmonella isolates recovered from retail chicken carcasses in six regions of China. The findings highlight the emergence of ciprofloxacin and cefotaxime co-resistant S. Indiana and also a mcr-1 positive S. Indiana with heavy multi drug resistance.
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Objective To investigate the effects of vaspin on insulin resistants of 3T3-L1 adipocyte through the insulin receptor substrates (IRS) /phosphatidylinositol 3-kinase (PI3K) /protein kinase B (Akt) /glucose transporter (Glut) signaling pathway.Methods 3T3-L1 cells cultured by palmitic acid (PA) were used to establish insulin resistance models,which were divided into PA group,PA + 100 ng/ml vaspin group,PA+200 ng/ml vaspin group,PA+400 ng/ml vaspin group and PA+400 ng/ml vaspin+wortmannin (PI3K inhibitor) group.Glucose uptake and consumption were assessed by 2-deoxy H3-D-glucose incorporation and glucose oxidase-peroxidase respectively.IRS/PI3K/Akt/Glut signaling pathway was evaluated using reverse transcription polymerase chain reaction and Western blot analysis.Results Compared with PA group,glucose uptake and consumption increased gradually with the increasing of vaspin concentration in other groups (P ＜ 0.05).mRNA levels of IRS-1,Akt and Glut 4 increased gradually as vaspin concentration increasing (P＜0.05),and the ratios of p-IRS-1 to IRS-1,p-Akt to Akt and Glut 4 protein level also showed the same trends (P＜0.05).However,glucose uptake and consumption in PA+400 ng/ml vaspin+wortmannin group were less than that of PA +400 ng/ml vaspin group (P＜0.05).PA+400 ng/ml vaspin+wortmannin group showed lower mRNA and protein phosphorylation levels of IRS-1,Akt and Glut 4 (P＜0.05),and that the ratios of p-IRS-1 to IRS-1,p-Akt to Akt and Glut 4 protein levels decreased (P＜0.05).Conclusions Vaspin can improve the insulin sensitivity of 3T3-L1 adipocyte by activating IRS/PI3K/Akt/Glut signaling pathway.
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Objective To identify the clinical features and risk factors of early rheumatoid arthritis (RA)-associated osteoporosis in premenopausal women. Methods A total of 76 premenopausal women with early RA were randomly selected in the Department of Kidney and Rheumatology in the hospital. A total of 84 health cases were randomly selected in our hospital as controls. Bone mineral density (BMD) was determined using dual energy X-ray absorptiometry (DEX). Bone metabolism (CTX, PINP) and inflammatory cytokines (IL-17, IL-6, TNF-α) were examined with quantitative enzyme-linked immune-sorbent assay (ELISA). Quantitative data were expressed as x ±s deviation and the data were compared between groups using non- parametric test (Z value). Multi-group comparison was performed with variance analysis. Qualitative data were compared with Fisher's test. Logistic regression was used to investigate the risk factors. Results ①Compared with the control group, BMD in the premenopausal women with early RA group [neck: (0.802 ±0.193) g/cm2, GT zone: (0.923±0.033) g/cm2, L1: (0.862±0.011) g/cm2] was significantly decreased [(0.981±0.032) g/cm2, (0.771 ±0.023) g/cm2, (0.912 ±0.012) g/cm2, F=14.401, 19.860, 6.560, respectively, both P<0.05). The prevalence of osteoporosis in this group was 7%(5/76), which was higher than controls 1%(1/84). ② According to values of Bone meta-bolism [(CTX: (0.37±0.21) ng/ml] and inflammatory cytokines, TNF-α: (9.8±4.1) pg/ml, IL-6: (33.6±5.7) pg/ml and IL-17: (129±24) pg/ml were markedly increased in premenopausal women in early RA group [(0.24 ±0.09) ng/ml, (6.7 ±1.9) pg/ml, (1.5 ±0.4) pg/ml, (45 ±7) pg/ml, Z=2.722, 5.932, 7.501, 4.370, respectively, both P<0.05]. ③ The premenopausal women with early RA group with osteoporosis were signifi- cantly difference with controls in BMI [(9±3) kg/m2 vs (16±3) kg/m2], bone density of neck [(0.85±0.20) ng/ml vs (0.88±0.14) g/cm2], L2 [(0.75±0.23) g/cm2 vs (0.88±0.14) g/cm2], L3 [(0.87±0.07) g/cm2 vs (0.93±0.14) g/cm2], L4 [(0.92±0.12) g/cm2 vs (0.94±0.16) g/cm2], serum ESR [47.8(22.0, 76.0) mm/1 h vs 18.8(8.7, 35.2) mm/1 h] and DAS28-CRP (5.3 ±1.2 vs 3.8 ±1.2) F=0.68, 14.632, 26.114, 20.931, 36.582, Z=3.21, 6.58, respectively, both P<0.05. ④ Logistic regression showed that IL-6 (Wald χ2=5.78, P=0.021), PINP (Wald χ2=5.12, P=0.031), CTX (Wald χ2=9.17, P=0.003), ESR (Wald χ2=9.24, P=0.011), DAS28-CRP (Wald χ2=17.28, P=0.001) were significantly positively correlated with osteoporosis. Moreover, ordered unconditional Logistic regression analysis of the variables (IL-6, PINP, CTX, ESR, DSA28) described above showed that DAS 28-CRP score [OR=1.58, 95%CI: (1.10, 2.20)] was the most important risk factor for osteoporosis in premenopausal women with early RA. Conclusion The incidence of osteoporosis is high in premenopausal women with early RA than healthy cases. DAS 28-CRP score is the important risk factor for premenopausal women with early RA- associated osteoporosis. Measures relieve symptoms of RA can help to prevent and treatment osteoporosis.
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Objective To explore the optimal time and sequence for getting the best magnetic resonance (MR) imaging when MR image of synovium macrophages was used for the diagnosis of collageninduced arthritis (CIA) in a rat model,and whether it can be used to monitor the efficacy of drug treatment.Methods CIA was induced by subcutaneous injection of chicken type Ⅱ collagen and complete Freund's adjuvant (CFA).Arthritis rats were randomly divided into the model group,the leflunomide group and the control group.Knees of the model group rats were imaged before and 24 h,48 h,72 h after USPIO intravenous administration (300 μmol Fe/kg) on day 28,29,30,31,respectively.From day 28,the leflunomide group was given a gauge of drug at a dose of 8 mg/kg.Then they were imaged before and 24 hours after USPIO administration on day 42,43 respectively.MR sequences included SE T1WI,SE T2WI,GRE T2 * WI.After the completion of MR imaging,rats were sacrificed to obtain histopathologic samples of synovial membrane.LSD-t test and one-way analysis of variance (ANOVA) were used for statistical analysis.Results No distinct signal enhancements were observed on the 24 h,48 h,72 h post-contrast enhancement on T1WI.On T2WI,signal intensity ratio of synovium (SNR) pre-contrast and 24 h,48 h post-contrast were 24.13±1.96,17.09± 1.23,19.14±0.91,respectively.On T2 * WI,SNR pre-contrast and 24 h,48 h post-contrast were 22.28±0.92,11.40±0.53,17.18±0.63,respectively.Distinct signal changes were observed on 24 h,48 h post-contrast on T2WI and T2 * WI (P＜0.05).The changes between SNR at 24 h,48 h,72 h post-contrast and pre-contrast were-29.09±2.42,-20.83±2.90,-6.2±2.9 respectively on T2WI,which were-48.4±1.3,-22.9±0.8,-8.2±1.6 respectively on T2 * WI.Changes were more obvious at 24 h post-contrast than 48 h post-contrast on both T2WI and T2 * WI (P＜0.05).The quantitative analyses were coinci-dent with the visual differences in signal changes between pre-contrast and post-contrast images.Difference between △SNR of leflunomide group and the control group on T1WI was not significant,while that on T2WI and T2 * WI were significantly different (P＜ 0.01).Histological examination confirmed the uptake of iron in the macrophages of arthritic knees.Signal intensity changed more on GRE T2 * WI than SE T2WI in all arthritis rats.Conclusion GRE T2 * WI is more sensitive for the diagnose of rat CIA,and 24 h post-contrast is better than 48 h and 72h post-contrast to get better images.We successfully observed the effects of leflunomide through signal changes of synovium,and the technique maybe contribute to diagnosis and therapeutic monito-ring of rheumatoid arthritis.
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Objective To investigate the protective effect of rhein lysinate (RHL)on the liver of the models with diabetic rats,and to provide basis for research on treatment of fatty liver in the patients with diabetes mellitus. Methods The models of diabetic rats were established by intraperitoneally injecting streptozotocin(STZ).40 rats were divided into control,model,25 mg·kg-1 RHL,and 50 mg·kg-1 RHL groups(n=10).The levels of malonaldehyde (MDA)and the activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) were detected by thiobarbituric acid method, pyrogallol autoxidation method, and NADPH coupling method, respectively.The pathological changes of liver tissue were observed by hematoxylin and eosin (HE)staining;the content of fat in liver tissue was observed by Nile red staining;the expression levels of fat synthesis-related proteins were detected by Western blotting method.Results Compared with control group,the body weight of the rats in model group was decreased and the levels of blood glucose,total cholesterol(TC)and triglyceride(TG)were increased (P<0.05);the activities of SOD and GSH-Px in liver tissue were decreased (P<0.05);there were a plenty of fat vacuoles and fat accumulation in liver tissue. The signal pathway of fat synthesis-related ERK1/2-SREBP-1c was activated in model group;compared with model group,it was inhibited in 25 and 50 mg· kg-1 RHL groups (P<0.05).Compared with model group,the blood glucose,TC and TG of the rats in 25 and 50 mg ·kg-1 RHL groups were decreased (P<0.05);the activities of SOD and GSH-Px were increased (P<0.05);however the body weight had no change. Compared with model group, the fatty vacuoles and the fatty accumulation of liver tissue in 25 and 50 mg·kg-1 RHL groups were decreased. Conclusion The hepatic protection of RHL is correlated with the inhibition of oxidative stress, fat degeneration and fatty accumulation of liver tissues.
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Objective To prospectively evaluate the ability of 7.0 T magnetic resonance (MR) imaging enhanced with ultrasmall superparamagnetic iron oxide (USPIO) particles in detecting synovial macrophages in a rat model of collagen-induced arthritis (CIA).Methods CIA was induced by subcutaneous injection of chicken type Ⅱ collagen.Arthritis rats were randomly divided into early,middle and late model group.Other rats served as controls.Knees of rats were imaged by a 7.0 T MR imager before and 24 hours after USPIO intravenous administration (200 μ.mol Fe/kg) on Day 14,28,42,respectively.The MR imaging protocol included T1-weighted (T1W) spin-echo (SE) sequences,T2-weighted (T2W) SE sequences,T2 * W gradientecho (GRE) sequences.After the completion of MR imaging,rats were sacrificed to obtain histopathologic samples of synovial membrane and staining.LSD-t test and one-way ANOVA were used for statistical analysis.Results Knees of arthritis rats in early,middle and late model groups developed synovial thickening and joint effusion on images from all sequences,whereas not developed in the control group.On USPIO-enhanced images,synovial tissue of arthritis knees showed slight to distinct signal intensity loss on post contrast T2WI and T2WI.However,no signal intensity enhancement of inflamed synovium was observed on postcontrast T1WI.Meanwhile,no control knees developed any visually detectable changes between pre-and postcontrast images.The quantitative analyses were coincident with the visual differences in signal changes between preand postcontrast images.Histologic examination confirmed uptake of iron in the macrophages of arthritic knees.Conclusion USPIO enhanced 7.0 T MR imaging can detect synovial macrophages in different period of CIA,which may be an additional tool for early and accurate diagnosis of RA.
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<p><b>OBJECTIVE</b>To investigate the effect of functional blocking of endogenous miR-23a with a specific antisense oligonucleotide (ASO) on the proliferation and invasiveness of gastric adenocarcinoma cell line MGC803 in vitro.</p><p><b>METHODS</b>A specific ASO targeting miR-23a, namely ASO-23a, was transfected into MGC803 cells to block endogenous miR-23a. The mRNA level of miR-23a in the transfected cells was detected with quantitative real-time PCR. The changes of cell proliferation following the transfection were detected with MTT assay and colony formation assay, and TUNEL assay and Transwell assay were employed to evaluate the changes in cell apoptosis and invasiveness, respectively.</p><p><b>RESULTS</b>Quantitative real-time PCR demonstrated efficient functional blocking of endogenous miR-23a in MGC803 cells by ASO-23a. Suppression of miR-23a with ASO-23a obviously inhibited cell growth, colony formation and invasiveness of MGC803 cells and significantly enhanced the cell apoptosis.</p><p><b>CONCLUSION</b>ASO-23a can efficiently block the function of endogenous miR-23a in MGC803 cells to inhibit cell proliferation and invasion and promote cell apoptosis.</p>
Subject(s)
Humans , Adenocarcinoma , Genetics , Pathology , Apoptosis , Cell Line, Tumor , Cell Proliferation , MicroRNAs , Genetics , Oligonucleotides, Antisense , Stomach Neoplasms , Genetics , Pathology , TransfectionABSTRACT
ObjectiveTo observe the effect of betulinic acid(BetA) on the growth of human cytokine induced killer(CIK) cells and the killing activity of CIK cells on the gastric cancer cells in vitro before and after induced by betulinic acid,explore its mechanism.MethodsPeripheral blood mononuclear cell (PBMC) were separated form the healthy and were induced with various of cytokine to become CIK cells in vitro.CIK cells were collected on the tenth day and were induced with betulinic acid in different concentrations,followed by 48 h,the colorimetric methyl thiazolyl tetrazolium(MTT) method assay the proliferation rate of human CIK cells.Flow cytometry (FCM) was used to detect the expression changes of perforin,granzyme B and CD107a of human CIK cells before and after betulinic acid-induced.Lactate dehydrogenase (LDH) release assay was used to measure the influence on cytotoxic activity of CIK cells induced by betulinic acid against gastric cancer cell line SGC-7901 in vitro.Western blot assay was used to measure the extracellular signal-regulated kinase1/2 (ERK1/2),and adapter proteins SH2-domain containing leukocyte protein of 76KD(SLP-76) and linker for activative of T cells(LAT) expression changes of human CIK cells before and after drug-induced.ResultsBetulinic acid can promote CIK cells growth when the concentration were in 0.08-10 μg/ml,the expression of perforin,granzyme B and CD107a of CIK cells were significantly higher than control group(P＜0.05) when the concentration of betulinic acid were in 0.3 μg/ml.In the meanwhile,the cytotoxic activity of CIK cells in vitro against gastric cancer cell line SGC-7901 were also remarkably higher than the control group (P＜0.05).The expression of SLP-76,LAT and ERK1/2 were significantly increased to a certain extent than the control group( P＜0.05 ),when CIK cells were treated with betulinic acid.ConclusionThese results suggest that betulinic acid can promote CIK cells growth in some concentrations and increase the cytotoxic activity of CIK cells against gastric cancer cell line SGC-7901,its mechanism may related with two factors,on the one hand,enhancing the activity of SLP-76,LAT and ERK1/2,on the other hand,increasing the expression of perforin,granzyme B and CD107a on the surface of CIK cells.
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ObjectiveTo investigate the association of complement C3 with clinical and serological features of patients with systemic lupus erythematosus.MethodsData was collected by the same methods in the past ten years in fifteen hospitals in Jiangsu Province and then data weres summarized for retrospective analysis.Clinical and laboratory data were selected and then analyzed by Chi-square test,Wilcoxon rank sum test and Logistic regression.ResultsOne thousand four hundred and five patients were investigated.One thousand and forty two had low serum complement C3 level.In this case control study,hospitalization age,disease course,admission times,pleurisy,gastrointestinal involvement,general lymphadenopathy/hepatosplenomegaly,white blood cell count, haemoglobin level,platelet count, serum C-reactive protein level,serum albumin level,serum creatinine level,Urine protein quantification,anti-nuclear antibodies (ANA),anti-dsDNAantibodies, anti-SmantibodiesandSLEDAIscore were possible factors associatedwith complement C3 reduction(P＜0.05).Logistic regression analysis showed that CRP (OR=0.396,0.254-0.617,P=0.000),ANA (OR=2.907,1.267-6.670,P=0.012),urine protein level(OR=1.702,1.043-2.779,P=0.033) and SLEDAI score (OR-0.930, 0.886-0.975,P-0.003) were correlated with complement C3 reduction.Conclusion Complement C3 level is valuable for lupus flare assessment.The complement C3 reduction is a risk factor for renal impairment.
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Objective To study the effect of 99Tc-MDP on receptor activator of nuclear factor-κB ligand/osteoprotegerin (RANKL/OPG) system of collagen-induced arthritis (CIA) rat. Methods The CIA was established by subcutaneous injection with type Ⅱ collagen and complete Freud's adjuvant to rats, except the group (n=8, treated with 99Tc-MDP). The histopathological changes of joints were scored based on the extent of infiltration of inflammatory cells, cartilage destruction and bone erosion. The serum levels of RANKL/OPG were tested with ELISA, and the synovial membrane levels of RANKL/OPG were tested with Western blot.Repeated ANOVA and one way ANOVA were used for statistical analysis. Results The MTX group relative value of RANKL/OPG in serum [35 d (1.076±0.016), 42 d (1.140±0.005), 49 d (1.155±0.023)], and in synovial membrane (1.156±0.014), and the histopathological scores [synovial membrane (1.8±0.5), cartilage (1.9±0.6), bone (1.8±0.5) ] were lower than model group ( 1.269±0.025, 1.296±0.015, 1.340±0.011 ), (2.9±0.4, 2.6±0.5, 2.6±0.5), ( 1.340±0.013 )(P＜0.01 ). And all above index of 99Tc-MDP group ( 1.035±0.034,0.986±0.019, 0.991±0.020), (1.5±0.5, 1.4±0.5, 1.2±0.5), (0.098±0.026) were notably lower than the MTX treat ment group (P＜0.01). Conclusion 99Tc-MDP may retard the progression of the disease by decreasing the relative RANKL/OPG expression and it can initiate its effect earlier than MTX.
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Objective To explore the relationship between the impairment of hematological system and disease activity,immunological parameters,and the prognosis of systemic lupus erythematosus (SLE).Methods The clinical and laboratory data of in-patients with SLE in Jiangsu Province were investigated and all patients were hospitalized between 1999-2009.The impairment of hematological system was assessed and the relationship between hematological system damage and disease activity,immunological parameters,mortality rate of patients with SLE were analyzed.Statistic method used was X2 test.Results One thousand nine hundred and fifty eight cases of SLE were included in the study,in which,1836 were female and 122 were male.One thousand five hundred and forty nine (79.1%) patients complicated with hematological system damage,62.3% were anemia,45.5% with leucopenia and 29.4% with thrombocytopenia.There were significant differences in hematological system damage rate among patients with mild activity group,moderate activity group,severe activity group and almost no activity group,compared respectively with almost no activity group.The P values were P=0.01 and P＜0.01 respectively.The incidence of hematological system damage in elevated ESR,low complement C3 level,anti-dsDNA antibody group was higher than that in patients who had normal ESR,complement C3 level and anti-dsDNA group.(P＜0.01).During follow-up,166 patients died,of which the mortality rate(91.6%) in patients had hematological system damage,was obviously higher than those without hematological damage(8.4%)(P＜0.01 ).Among the 166 deceased patients,38.6% died of severe infection,22.9% died ofrenal failure,15.1% died ofnervous system damage,10.2% died of cadiovascular damage and 13.3% died from other causes.Conclusion Hematological system is one of the most commonly involved system in patients with SLE,of which anemia is the most common,and the incidence of leukopenia follows.The impairment of hematological system is closely related to lupus activity.Patients with abnormal immune parameters tend to have secondary hematological system damage.Severe infection is the main cause of death in patients with lupus,followed by nervous system damage and kidney damage.The mortality rate in patients with lupus that complicated hematological system damage is higher than patients who have no hematological system damage.
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Objective To investigate the initial manifestation and disease onset feature of systemic lupus erythematosus(SLE) in the past ten years in fifteen hospitals in Jiangsu Province.Methods Data was collected by the same Methodsin all the participated hospitals and then it was summarized for retrospective analysis.Two groups were compared by chi-square test.Results ① One thousand nine hundred and fifty eight patients were investigated and the male-to-female ratio was 1∶15.0.② One thousand seven hundred and ninty eight patients had clear initial manifestations.The most common initial manifestations were skin and mucosal lesions(769 cases,42.8％ ) and arthritis (697 cases,38.8％ ).The main skin lesion was malar rash (706 cases).Arthritis was found to be more common in female than male.③ All hospitalized patients at their first admission showed multiple organ/system involvement:the most common involvement was skin and mucous membrane (82.3％),hematologic damage (74.0％),in which at least one series of blood cells were involved,arthritis (1156 cases,56.5％ ) much more than myositis (51 cases),proteinuria 1046 cases and hematuria in 385 cases.Renal biopsy pathology showed type Ⅳ glomerulonephritis.Conclusion ① SLE patients are mainly female and the male to female ratio is 1∶15.0.② The most common initial manifestations are skin and mucosal lesions.③ The most commonly involved organ/system are skin and mucous membrane,blood,joint and kidney.The most common pathological changes shown in renal biopsy is type Ⅳ glomerulonephritis.
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This study was aimed to detect the expression of c-Jun N-terminal kinase (JNK) in renal tissues of lupus nephritis (LN) mice with chronic graft-versus-host disease (cGVHD) and to investigate that LN could be intervened by fluvastatin at different doses through the inhibition of JNK expression.LN models with cGVHD in mice were established first,and then diseased mice were randomly divided into four groups:normal control,fluvastatin intervention at high-dose group (10 mg/kg),fluvastatin intervention at low-dose group (5 mg/kg) and models without treatment.After killing the mice sixteen weeks later,total urine protein of every mouse was determined by a Biuret colorimetric assay.The protein and mRNA levels of JNK and p-JNK in kidneys were measured by immunohistochemistry,Western blot and RT-PCR,respectively.Compared with normal control,otal urine protein,JNK and p-JNK expressions at both mRNA and protein levels were significantly increased in cGVHD group (P ＜0.01),but their expressions were suppressed by fluvastatin treatment (P ＜0.01).JNK may play an important role in the pathogenetic progress of LN in mice,and fluvastatin is able to prevent LN via inhibition of JNK expression in renal tissues in cGVHD mice.
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Objective To investigate the genotypes and encoding resistance genes differences of Acinetobacter baumannii and analyze their interrelations with multi-drug resistance.Methods A total of 77strains Acinetobacter baumannii were collected random from the second Xiangya Hospital during September 2008 to September 2009.The K-B method which was WHO recommended was adopted to Acinetobacter baumannii drug sensitivity test to 15 kinds of antibiotics to establish susceptibility spectrum.At the same time,random amplified polymorphic DNA(RAPD)technique was used to establish DNA fingerprinting.The genes of β-lactamase(TEM-1,IMP,OXA-23,OXA-24,AmpC),aminoglycoside-modifying enzymes[aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ]and 16S rRNA methylase(armA,rmtA,rmtB)were detected by PCR and sequenced,and find the relationship between the gene encoding and multi-drug resistance.In addition,we compared the rates of resistance genes of Acinetobacter baumannii and the relations with the genotype and the multi-resistance.Results Thirty-one sensitive strains and 46 multi-drug resistance strains(10 Pan-drug resistances)were isolated.Seventeen types from A to Q were separated using RAPD technique.E genotype widely popular in the ICU was the advantage type in multi-drug resistance strains,and the rate was 47.1%.While the various types scattered in sensitive strains.The positive rates of TEM-1,IMP,OXA-23,OXA-24,AmpC,aac(3')-Ⅰ ,aac(6')-Ⅰ ,ant(3")-Ⅰ ,armA in the multi-drug resistance strains and the sensitive strains were 95.7%,39.1%,84.8% ,54.3%,87.0%,89.1%,84.8%,45.7%,63.0% and 58.1%,9.7%,32.3%,48.4%,48.4%,29.0%,45.2%,12.9%,9.7%,respectively,and there was significant difference except for OXA-24 using the X2 test(P ＜ 0.05).All isolates were negative for rmtA gene and rmtB gene.Drug susceptibility analysis showed that the resistant rate was significantly higher of the strains carrying resistant genes than that of the resistance negative strains.When the strains were resistant to gentamicin and amikacin,the rate of three aminoglycoside genes positive was 34.8%.The trains containing all the measured β-lactamase genes were all resistant strains.Conclusion Compared with the sensitive Acinetobacter baumannii strains,a broad resistance spectrum and a high drug resistance rate were showed in multidrug resistance strains isolated from clinic,which harboring many kinds of β-lactamase genes and aminoglycosides genes with a high separation rate,and the same clone of multiple drug-resistant strains may be transmitted in and among wards.
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AIM: To investigate the relationship between TGF-?_1 over-expression and lupus nephritis(LN) in BXSB mice and whether MPS ameliorates LN by inhibiting the over-expression of TGF-?_1. METHODS: BXSB mice (experiment group, n=6) were treated (i.p.) with MPS (25 (mg?kg~(-1)?d~(-1))) dissolved in N.S for 3 weeks. The other age and sex-matched BXSB mice (n=6) and BALB/c mice (n=6) received N.S. alone. Proteinuria production was evaluated once a week. The expression of TGF-?_1 in the kidney was investigated by means of immunohistochemistry and RT-PCR. RESULTS: MPS significantly reduced proteinuria of BXSB mice (P
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Objective To explore whether inducible nitric oxide synthase(iNOS) could mediate lupus nephritis(LN) in BXSB mice and whether methyl prednisolone ameliorates LN by inhibiting iNOS expression.Methods Eighteen week old male BXSB mice (n=6) that had clinical diagnosis of glomerulonephritis were treated(i.p.) for 3 weeks with methylprednisolone(25 mg?kg-1?day-1).Controls were age- and sex-matched BXSB mice (n=6) that received normal saline only.Age- and sex- matched BALB/C mice (n=6) were used as normal controls that were treated in the same way as the BXSB controls.iNOS expression was assessed by RT-PCR and immunochemistry.Proteinuria and urinary nitrite/nitrate production were also evaluated.Results iNOS was strongly expressed in kidneys of all BXSB mice,while no signal of expression was found in kidneys of BALB/C mice by both RT-PCR and immunochemistry.Methylprednisolone reduced iNOS expression(P
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Objective To investigate interleukin 10 (IL 10,Th2 type cytokines) and interferon gamma (IFN ?,Th1 type cytokines) secreting capacity by peripheral blood mononuclear cells (PBMCs) in 30 patients with systemic lupus erythematousus (SLE) and 10 healthy individuals,and try to understand the role of Th1/Th2 balance in the pathogenesis of SLE.Method Sandwich ELISA was used to measure IL 10 and IFN ? level in the supernatant of culture from PBMCs.Results The level of IL 10 produced spontaneously by PBMCs in active stage was significantly higher than in static phase and healthy person ( P 0 05).Conclusion These data suggest that the Th2 type cytokine (IL 10) was increased in patients with SLE,and an imbalance towards Th2 predominance may play a significant role in the pathogenesis of SLE.Regulating IL 10 level may provide a new approach for the treatment of SLE.
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Objective To investigate whether monocyte chemoattractant protein-1 mediates lupus nephritis (LN) in BXSB mice and whether methylprednisolone ameliorates LN by inhibiting MCP-1 expression. Methods 18-week-old male BXSB mice (n=6) displaying clinical symptoms of glomerulonephritis were treated (i.p.) for 3 weeks with MPS (25 mg?kg-1?d-1) dissolved in N.S. BXSB controls were age-and sex-matched BXSB mice (n=6) that received N.S. alone. Age- and sex-matched BALB/C mice (n=6) were used as normal controls that were treated in the same way as the BXSB controls. MCP-1 expression was investigated by RT-PCR and immunohistochemical examination. The heaviness of proteinuria was also evaluated. Medical imaging analysis was performed to detect the relationship between MCP-1 expression and proteinuria. Results MCP-1 was strongly expressed in kidneys of all BXSB mice, stronger staining was found in cytoplasm of tubular epithelial cells than glomerular, while no expression was found in kidneys of BALB/C mice. MCP-1 expression in tubular cells in BXSB control group was closely correlated with proteinuria. MPS significantly reduced proteinuria and MCP-1 expression and down-regulation of MCP-1 expression in tubular cells was also closely correlated with that of proteinuria. Conclusion MCP-1 over-expression may mediate LN in BXSB. MPS ameliorates LN partially by inhibiting MCP-1 expression.