Evaluation of the efficiency of ciprofloxacin against S. Typhi by altering the production of cytokines in acute typhoid fever in patients at Al-Diwaniyah Hospitals, Iraq

Aims@#Typhoid fever is a life-threatening disease in the developing world that claims >600,000 deaths per year. Its causative agent Salmonella Typhimurium (S. Typhi) can be treated with ciprofloxacin, an effective broad-spectrum antibiotic that enhances the natural host defenses. However, the emergence of resistant bacterial strains may be a warning alarm against the clinical use of this antibiotic. This study was aimed to investigate the efficiency of ciprofloxacin treatment (250 mg/mL) against S. Typhi by altering the production of serum cytokines IL-10, 1L-6 and TNF-α in acute typhoid fever patients in Diwanyah Hospitals.@*Methodology and results@#ELISA and Western Blot methods were used to investigate cytokine levels in patients and healthy controls sera. Our results showed that all cytokines’ levels before treatment with ciprofloxacin were significantly higher than the control (healthy group). However, treated patients with ciprofloxacin revealed a significantly reduced concentration of IL-10 and TNF-α compared to untreated control samples. However, the level of IL-6 was higher even with ciprofloxacin treatment.@*Conclusion, significance and impact of study@#The study concluded that ciprofloxacin (250 mg/mL) might significantly alter serum cytokines IL-6, IL-10 and TNF-α levels in acute typhoid fever patients. Therefore, further molecular studies are essential to understand the effect of ciprofloxacin on the production of cytokines.

Liraglutide ameliorates palmitic acid-induced insulin resistance in L6 skeletal muscle cells by regulating Sestrin2 / 中华内分泌代谢杂志

Objective:To investigate the role of stress-inducible protein Sestrin2 (Sesn2) in the improvement of insulin resistance in rat L6 skeletal muscle cells treated with liraglutide.Methods:The establishment of insulin resistance model of rat L6 skeletal muscle cells was induced by palmitate. The experimental cells were divided into control group(Con group), palmitate 0.6 mmol/L treatment group(PA group), palmitate 0.6 mmol/L+ liraglutide 10 nmol/L treatment group(PA+ Lir10 group), palmitate 0.6 mmol/L+ liraglutide 100 nmol/L treatment group(PA+ Lir100 group), and palmitate 0.6 mmol/L+ liraglutide 1 000 nmol/L treatment group(PA+ Lir1000 group). The cell counting kit 8(CCK8) method was used to detect the cell activity in each group. Western blotting was used to detect the expression levels of glucose transporter 4(GLUT4), protein kinase B(Akt), phosphorylated protein kinase B(p-Akt), and Sesn2 protein in L6 cells. L6 cells were transfected with siRNA to inhibit the expression of Sesn2. The cells were treated with palmitate and liraglutide. Western blotting was used to detect the expression levels of Sesn2, Akt, p-Akt, and GLUT4 protein in L6 cells.Results:Compared with Con group, the cell survival rate, p-Akt/Akt ratio, Sesn2, and GLUT4 protein expression in PA group decreased significantly( P<0.05). After liraglutide intervention, the cell activity, p-Akt/Akt ratio, Sesn2, and GLUT4 protein expression of PA+ Lir100 and PA+ Lir1000 groups was increased( P<0.05). After inhibiting the expression of Sesn2, p-Akt/Akt ratio and GLUT4 protein in transfected si-Sesn2 and treated with 0.6 mmol/L palmitate group(PA+ si-Sesn2 group) and transfected si-Sesn2 and treated with 0.6 mmol/L palmitate+ liraglutide 100 nmol/L group (Lir100+ PA+ si-Sesn2 group) were significantly lower than those in transfection negative group (si-Con group; P<0.05). Even after liraglutide intervention, compared with PA+ si-Sesn2 group, p-Akt/Akt ratio and GLUT4 protein expression level were not significantly increased in Lir100+ PA+ si-Sesn2 group ( P>0.05). Conclusions:Palmitate could induce the decrease of p-Akt/Akt ratio and GLUT4 protein expression in L6 cells. Liraglutide upregulates the expression of Sesn2, which leads to the increase of p-Akt/Akt ratio and GLUT4 protein expression and contributes to the improvement of insulin resistance.

Using PacBio sequencing to investigate the effects of treatment with lactic acid bacteria or antibiotics on cow endometritis

BACKGROUND: Endometritis is the most common disease of dairy cows and traditionally treated with antibiotics. Lactic acid bacteria can inhibit the growth of pathogens and also have potential for treatment of endometritis. Using PacBio single-molecule real-time sequencing technology, we sequenced the fulllength l6S rRNA of the microbiota in uterine mucus samples from 31 cows with endometritis, treated with lactic acid bacteria (experimental [E] group) and antibiotics (control [C] group) separately. Microbiota profiles taken before and after treatment were compared. RESULTS: After both treatments, bacterial species richness was significantly higher than before, but there was no significant difference in bacterial diversity. Abundance of some bacteria increased after both lactic acid bacteria and antibiotic treatment: Lactobacillus helveticus, Lactococcus lactis, Lactococcus raffinolactis, Pseudomonas alcaligenes and Pseudomonas veronii. The bacterial species that significantly decreased in abundance varied depending on whether the cows had been treated with lactic acid bacteria or antibiotics. Abundance of Staphylococcus equorum and Treponema brennaborense increased after lactic acid bacteria treatment but decreased after antibiotic treatment. According to COG-based functional metagenomic predictions, 384 functional proteins were significantly differently expressed after treatment. E and C group protein expression pathways were significantly higher than before treatment (p < 0.05). CONCLUSIONS: In this study, we found that lactic acid bacteria could cure endometritis and restore a normal physiological state, while avoiding the disadvantages of antibiotic treatment, such as the reductions in abundance of beneficial microbiota. This suggests that lactic acid bacteria treatment has potential as an alternative to antibiotics in the treatment of endometritis in cattle.

Antihyperglycemic and antihyperlipidemic activities of wild musk melon (Cucumis melo var. agrestis) in streptozotocin-nicotinamide induced diabetic rats / 中草药·英文版

Objective: Wild musk melon (Cucumis melo var. agrestis, CMA) is one of the edible plants form Tamil Nadu. Traditionally, this plant was used as diabetic diet (leaves of CMA with Momordica charantia leaves), but there is no scientific report on antidiabetic action of this plant material. Hence, the current research work was designed to evaluate the antihyperglycemic and antihyperlipidemic effect of hydroalcoholic extract of CMA leaves (HALEC) in streptozotocin (STZ)-nicotinamide (NIC)-induced diabetic rats. Methods: Diabetes was induced by administration of STZ (60 mg/kg, i.p.) after 15 min of NIC (120 mg/kg i.p.) administration. The diabetic rats were treated with HALEC (300 and 600 mg/kg, p.o., respectively) for 21 d. Results: After the management with HALEC, blood glucose, HbA1c levels, total cholesterol, LDL cholesterol, triglycerides levels, glycogen phosphorylase and glucose-6-phosphatase levels were significantly diminished in diabetic rats. However, haemoglobin level, HDL cholesterol, liver glycogen, total protein, hexokinase, glucose-6-phosphate dehydrogenase levels were significantly increased in HALEC treated diabetic rats. The histopathological studies of the pancreas in HALEC-treated diabetic rats showed almost normal appearance. L6 cell line study revealed the increased glucose uptake activity of HALEC. High performance thin layer chromatography (HPTLC) analysis confirms the presence of active principles such as rutin, gallic acid and quercetin in HALEC. Conclusion: The results indicated that HALEC possess significant antihyperglycemic and antihyperlipidemic activity in STZ-NIC-induced type II diabetic rats with protective effect. This research work will be useful for the isolation of active principles and development of herbal formulation in phytopharmaceuticals.

Action Mechanism of Main Components from Glycyrrhizae Radix et Rhizoma in Improving Insulin Resistance in L6 Rat Myoblasts / 中国实验方剂学杂志

Objective::To determine whether the main components of Glycyrrhizae Radix et Rhizoma can improve insulin resistance by regulating glycogen synthesis, glycolysis pathway and fatty acid synthesis in myoblasts of L6 rat myoblasts. Method::Insulin resistance (IR) model of L6 rat myoblasts was established through incubation with 0.05 mmol·L-1 palmitic acid (PA) for 9 hours. Normal group, model group, glycyrrhizic acid (GA, 25 μmol·L-1) group, 18β-glycyrrhetinic acid (18β-GA, 25 μmol·L-1) group, isoliquiritigenin (ILG, 25 μmol·L-1) group and isoliquiritin (ILQ, 25 μmol·L-1) group were set up, glucose content in supernatant of cell culture medium was detected by glucose kit, myoblasts glycogen content was determined by glycogen detection kit, protein expression levels of Sterol regulatory element binding protein-1c(SREBP-1c), fatty acid synthetase (FAS) and glycogen synthase kinase3β(GSK3β) were detected by Western blot, and the mRNA expressions of key enzymes in glycolysis were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR). Result::Compared with those in the normal group, the glucose consumption rate was significantly down-regulated in model group (P<0.01), the glycogen content was decreased (P<0.05), the protein expressions of Sterol regulatory element binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) were decreased (P<0.05, P<0.01), the mRNA expressions of fructose phosphate kinase 1 (PFK1), pyruvate kinase (PK) and hexokinase (HK) were down-regulated (P<0.05), and the protein expression of glycogen synthase kinase 3 (GSK3β) protein was increased (P<0.05). Compared with model group, GA, 18β-GA and ILG could significantly increase glycogen content in myoblasts of IR-L6 rats (P<0.05, P<0.01). GA, 18β-GA and ILQ could significantly increase the expression of SREBP-1c (P<0.05, P<0.01), and GA, 18β-GA, ILG and ILQ could significantly increase the expression of FAS (P<0.05, P<0.01), the mRNA expressions of PFK1, PK and HK (P<0.05, P<0.01), and down-regulate the protein expression of GSK3β (P<0.05). Conclusion::The main components of licorice improve the insulin resistance by promoting glycolysis and glycogen synthesis and regulating fatty acid synthesis.

Protective effect of epithelial growth factor on oxidative damage of skeletal muscle cells in model rats of oxygen-glucose deprivation / 吉林大学学报(医学版)

To set up the rat skeletal muscle L6 cell models of oxygen-glucose deprivation (OGD) in vitro, and to investigate the protective effect of EGF in deep tissue injury (DTI) of pressure sores. Methods: The rat skeletal muscle cells in the logarithmic phase were divided into normal control group, OGD group, 5 μg · L1 EGF+OGD group, 10 μg · L1 EGF+ OGD group and 20 μg · L1 EGF+ OGD group. The survival rates of skeletal muscle cells in various groups were measured by MTT assay; the cell apoptotic rates in various groups were detected by flow cytometry; the reactive oxygen species (ROS) levels were detected by DCFH-DA; Rhodamine 123 was used to detect the mitochondrial membrane potential; the expressions of Bax and Bel-2 proteins were determined by Western blotting method. Results: Compared with normal control group, the survival rates of skeletal muscle cells in OGD group after 24 h OGD was significantly decreased (P<0.05); the apoptotic rate was markedly increased (P<0.01); the ROS level was increased (P<0.01); the mitochondrial membrane potential was decreased (P<0.01); the ratio of Bcl-2/Bax was significantly decreased (P<0.01). Compared with OGD group, the survival rates of skeletal muscle cells in different concentrations of EGF groups were increased and the apoptotic rates were decreased, especially in 10 and 20 μg · L1 EGF groups (P<0.05 or P<0.01); the ROS levels in skeletal muscle cells in different concentrations of EGF groups were decreased and the mitochondrial membrane potential were increased, especially in 10 and 20 μg · L1EGF groups (P<0.05 or P<0.01); the Bcl-2/Bax ratios were significantly decreased in a concentration-dependent manner, especially in 10 and 20 μg · L1 EGF groups (P<0.05 or P<0.01). Conclusion: EGF can improve the skeletal muscle cell injury induced by OGD in a concentration-dependent manner via decreasing the ROS levels and protecting the cell mitochondrial function.

Protective effect of epithelial growth factor on oxidative damage of skeletal muscle cells in model rats of oxygen-glucose deprivation / 吉林大学学报(医学版)

Objective:To set up the rat skeletal muscle L6 cell models of oxygen-glucose deprivation(OGD)in vitro,and to investigate the protective effect of EGF in deep tissue inj ury(DTI)of pressure sores.Methods:The rat skeletal muscle cells in the logarithmic phase were divided into normal control group,OGD group,5 μg·L-1 EGF+OGD group,10 μg·L-1EGF+ OGD group and 20 μg·L-1EGF+ OGD group.The survival rates of skeletal muscle cells in various groups were measured by MTT assay;the cell apoptotic rates in various groups were detected by flow cytometry;the reactive oxygen species(ROS)levels were detected by DCFH-DA;Rhodamine 123 was used to detect the mitochondrial membrane potential;the expressions of Bax and Bcl-2 proteins were determined by Western blotting method.Results:Compared with normal control group,the survival rates of skeletal muscle cells in OGD group after 24 h OGD was significantly decreased(P<0.05);the apoptotic rate was markedly increased(P<0.01);the ROS level was increased(P<0.01);the mitochondrial membrane potential was decreased(P<0.01);the ratio of Bcl-2/Bax was significantly decreased(P<0.01).Compared with OGD group, the survival rates of skeletal muscle cells in different concentrations of EGF groups were increased and the apoptotic rates were decreased,especially in 10 and 20 μg·L-1EGF groups(P<0.05 or P<0.01);the ROS levels in skeletal muscle cells in different concentrations of EGF groups were decreased and the mitochondrial membrane potential were increased,especially in 10 and 20 μg·L-1EGF groups(P<0.05 or P<0.01);the Bcl-2/Bax ratios were significantly decreased in a concentration-dependent manner,especially in 10 and 20 μg·L-1EGF groups (P<0.05 or P<0.01).Conclusion:EGF can improve the skeletal muscle cell injury induced by OGD in a concentration-dependent manner via decreasing the ROS levels and protecting the cell mitochondrial function.

Sarilumab:A New Arsenal for Rheumatoid Arthritis

Rheumatoid arthritis is a debilitating disease that has significant impact on quality of life. Biological and non biological DMARDs improved the outcome in RA patients. New agents are still required in inadequate responders or intolerant patients. FDA has recently approved a new drug Sarilumab, a monoclonal antibody for the treatment of moderate to severe form of RA which acts by binding to Interleukin receptor 6 and interrupts the resultant cytokine-mediated inflammatory signaling.

Analysis on chemical constituents from Swertia mileensis by UPLC-ESI-Q-TOF-MS / 中草药

Objective: To analyze and identify the chemical constituents from Swertia mileensis by UPLC-ESI-Q-TOF-MS. Methods: The analysis was performed on an Acquity HSS T3 reverse phase column (100 mm × 2.1 mm, 1.8 μm). The mobile phase consisted of 0.1% formic acid acetonitrile and 0.1% formic acid, and was used for gradient elution, with the flow rate of 0.4 mL/min. Mass spectrometry was applied for the qualitative analysis under positive and negative ion modes and ESI ion source. Data was analyzed by Masslynx 4.1 software, SciFinder database, literatures, and standards. Results: Twenty-eight compounds, including 7 iridoids, 14 xanthones, 3 flavonoids, 2 triterpenes, and 2 phenols were identified from S. mileensis. Among them, vogeloside, 8-O-β-D- glueopyranosyl-(l→6)-β-D-glueopyranosyl-l,7-dihydroxy-3-ethoxyxanthone, 3-O-demethylswertipunicoside, and sweriyunnanlactone A were reported from this species for the first time. Conclusion: Using UPLC-ESI-Q-TOF-MS method the main chemical constituents from S. mileensis can be rapidly and accurately identified, which provides a new strategy for its quality control as well as a reference for clarifying the material basis of its efficacy.

Effect of Bcl-2 and PCNA expression in rat myoblast injury induced by hydrogen peroxide / 中国病理生理杂志

AIM:To explore the role of Bcl-2 and PCNA expression in the injury of rat myoblasts induced by hydrogen peroxide (H2O2).METHODS:Rat myoblasts at growth phase were divided into 4 groups based on basic fibroblast growth factor (bFGF) and H2O2 levels:normal control group, bFGF group, model group (H2O2 group) and treatment group (bFGF+H2O2 group).The expression of Bcl-2 and Bax was observed by immunohistochemistry and fluorescence methods.The protein levels of Bax, Bcl-2 and PCNA were determined by Western blot.RESULTS:Compared with model group, both immunofuorescence and fluorescence in treatment group showed enhanced Bcl-2 and low expression of Bax.Furthermore, the results of Western blot showed up-regulated PCNA and Bcl-2 protein and decreased Bax expression in treatment group.CONCLUSION:Oxidative stress results in the pathologic changes of myoblasts, and the up-regulation of Bcl-2 and PCNA may attenuate myoblast injury.

Exosomes Secreted by Toxoplasma gondii-Infected L6 Cells: Their Effects on Host Cell Proliferation and Cell Cycle Changes

Toxoplasma gondii infection induces alteration of the host cell cycle and cell proliferation. These changes are not only seen in directly invaded host cells but also in neighboring cells. We tried to identify whether this alteration can be mediated by exosomes secreted by T. gondii-infected host cells. L6 cells, a rat myoblast cell line, and RH strain of T. gondii were selected for this study. L6 cells were infected with or without T. gondii to isolate exosomes. The cellular growth patterns were identified by cell counting with trypan blue under confocal microscopy, and cell cycle changes were investigated by flow cytometry. L6 cells infected with T. gondii showed decreased proliferation compared to uninfected L6 cells and revealed a tendency to stay at S or G2/M cell phase. The treatment of exosomes isolated from T. gondii-infected cells showed attenuation of cell proliferation and slight enhancement of S phase in L6 cells. The cell cycle alteration was not as obvious as reduction of the cell proliferation by the exosome treatment. These changes were transient and disappeared at 48 hr after the exosome treatment. Microarray analysis and web-based tools indicated that various exosomal miRNAs were crucial for the regulation of target genes related to cell proliferation. Collectively, our study demonstrated that the exosomes originating from T. gondii could change the host cell proliferation and alter the host cell cycle.

Improved effect of curcumin on mitochondrial dysfunction induced by high glucose in L6 cells / 中国药科大学学报

@#To investigate the effects of curcumin on mitochondrial dysfunction induced by high glucose(40 mmol/L glucose, 24 h)in L6 cells, curcumin(10, 20, 40 μmol/L)was administered for 24 h after high glucose culture. The effects of curcumin on the mitochondrial dysfunction were evaluated by mitochondrial membrane potential, reactive oxygen species(ROS), ATP content and mtDNA copy number. The mRNA and protein expression of uncoupling protein 2(UCP2), PPARγ coactivator 1α(PGC-1α)and sirtuin-1(Sirt3)were also determined. As improvement of high glucose damage, curcumin significantly raised mitochondrial membrane potential and ATP content, and decreased ROS level. Curcumin significantly ameliorated the down regulation of UCP2 yet with little effect on mtDNA copy number and PGC-1α and Sirt3 expression. In conclusion, curcumin could significantly ameliorate mitochondrial dysfunction in L6 cells induced by high glucose, which involved the mechanism of multiple antioxidants.

Expression and clinical significance of ribosomal protein L6 in prostate cancer / 天津医药

Objective To explore the expression of ribosomal protein L6 (RPL6) in prostate cancer and its clinical sig-nificance. Methods RT-qPCR and Western blot assay were used to measure the mRNA transcription and protein expres-sion levels of RPL6 in prostate cancer tissues (n=80) and adjacent non-cancerous tissues (n=62). The relationship between RPL6 mRNA expression level and clinicopathological factors of prostate cancer was statistically analyzed. Results The mRNA and protein expression levels of RPL6 were significantly higher in prostate cancer tissues compared with those of non-cancerous tissues (P0.05). Kaplan-Meier survival analysis of biochemical recurrence (BCR)-free survival time showed the significantly lower recurrent rate in patients with high RPL 6 mRNA expression(χ2=4.530,P=0.033). Conclusion The elevated expression of RPL6 may play a role in the development of prostate cancer, and which can be used as a tumor marker to assess the prognosis of prostate cancer.

Screening of Active Fractions for Purple Sweet Potato Flavonoids with Insulin Resistance Improving Ability of L6 Cells / 中国药师

Objective:To screen the active fractions from purple sweet potatoes containing flavonoids with insulin resistance impro -ving ability.Methods:L6 cell model with insulin resistance was established .The extracting solutions with different polarity of purple sweet potato flavonoids were used to affect the model .The residual glucose concentration in insulin resistant L 6 cells was observed and compared before and after the intervention .Results:The residual glucose concentration of total flavonoids extracting solution , butanol extracting solution at middle and high dose , and chloroform extracting solution at all doses of purple sweet potato were lower than that of IR group (P<0.05).Conclusion:The total flavonoids extract, chloroform extracting solution and butanol extracting solution of purple sweet potato can improve insulin resistance in L 6 cells.

Methanolic extract of Momordica cymbalaria enhances glucose uptake in L6 myotubes in vitro by up-regulating PPAR-γ and GLUT-4 / 中国天然药物

The present study was undertaken to evaluate the influence of the methanolic fruit extract of Momordica cymbalaria (MFMC) on PPARγ (Peroxisome Proliferator Activated Receptor gamma) and GLUT-4 (Glucose transporter-4) with respect to glucose transport. Various concentrations of MFMC ranging from 62.5 to 500 μg·mL(-1) were evaluated for glucose uptake activity in vitro using L6 myotubes, rosiglitazone was used as a reference standard. The MFMC showed significant and dose-dependent increase in glucose uptake at the tested concentrations, further, the glucose uptake activity of MFMC (500 μg·mL(-1)) was comparable with rosigilitazone. Furthermore, MFMC has shown up-regulation of GLUT-4 and PPARγ gene expressions in L6 myotubes. In addition, the MFMC when incubated along with cycloheximide (CHX), which is a protein synthesis inhibitor, has shown complete blockade of glucose uptake. This indicates that new protein synthesis is required for increased GLUT-4 translocation. In conclusion, these findings suggest that MFMC is enhancing the glucose uptake significantly and dose dependently through the enhanced expression of PPARγ and GLUT-4 in vitro.

RNA interference affects the feak-headbox 3a gene expression in myoblast cell line L6 / 中国组织工程研究

BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy. OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro. METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours. RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (Phours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (Psignificantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.

Effects of angiotensin Ⅱ on insulin signal transduction in myoblasts of L6 rats / 中华内分泌外科杂志

Objective To study the effects of angiotensin Ⅱ (AngⅡ) on insulin signal transduction pathway in skeletal myoblast of L6 rats,and further to explore the possible mechanism of AngⅡ on glucose utilization.Methods Myoblast cells of L6 rats were cultured and induced to differentiate.They were divided into 4 groups according to different treatment by AngⅡ or JAK2-PKA inhibitor H89:normal control group ( NC group),insulin group,insulin + AngⅡ group and insulin + AngⅡ + H89 group.Expression of IRS1 and GLUT4 mRNA was detected by RT-PCR.Expression of IRS1,Ptyr-IRS1 and GLUT4 (total and membrane protein) were detected by Western blot.Results The difference of GLUT4 mRNA expression in the 4 groups detected by RT-PCR had no statistical significance(P ＞ 0.05).The difference of IRS1 mRNA expression among the latter 3 groups had no statistical significance(P ＞ 0.05),however,IRS1 expression in the latter 3 groups was higher than that in NC group(P ＜ 0.05).Western blot results showed expression of IRS1,Ptyr-IRS1 and GLUT4 (membrane protein)was higher in the latter 3 groups than in NC group(P ＜0.05).The difference of IRS1 expression among the latter 3 groups(P ＞ 0.05 ) and GLUT4 (total protein) expression among the 4 groups had no statistical significance (P ＞ 0.05).The expression of of ptyr-IRS1 and GLUT4 membrane protein in Ins + AngⅡ + H89 group was much higher than that in Ins + AngⅡ group,and lower than that in insulin group(P ＜0.05).Conclusion AngⅡ inhibits IRS1's tyrosine phosphorylation and GLUT4's transfer from cytoplasm to plasma membrane in skeletal muscle cells through JAK2-PKA signaling pathway,and therefore induces insulin resistance.

In vitro glucose uptake activity of Tinospora crispa in skeletal muscle cells

Background: In South Asian countries, stems of the plant, Tinnospora crispa, Linn (TC) are often used as a folk medicine in the treatment of type 2 diabetes mellitus. Though TC’s antiglycemic activity has been demonstrated in diabetic rats, the mechanism for its action has not yet been elucidated. Objective: To investigate the effect of TC’s aqueous extract (TCA) on glucose transport activity in skeletal muscle cell line. Materials and methods: A skeletal muscle cell line, L6 myoblasts, was used for this study. The myoblasts grown to the stage of fused myotubes were pre-incubated with and without TCA for 24 hours. Then, a 2-[³H]-deoxy-D-glucose (2-DG) uptake test was made in a 24-well plate for 10 minutes. In the downstream transport regulation studies, the TCA pre-incubated cells was either treated or untreated with specific inhibitors of the PI3-Kinase (wortmannin) and p38 MAP-Kinase (SB203580) pathways prior to the uptake test. All studies were carried out in triplicate with a minimum of three independent experiments. Results were expressed as mean±SE and compared with student’s t test for a level of significance at p \< 0.05. Results: TCA at 4 mg/mL significantly enhances glucose uptake of L6 myotubes in dose and time dependent manner with the half maximum effects at 24 hours (196.60±11.09%, p \< 0.05). The effect was completely abolished by a cytoskeletal blockade (10 μM of cytochalasin B), supportive of active glucose transport activity. Both wortmannin and SB203580 have no effect on the TCA-stimulated glucose uptake. Conclusion: Tinospora crispa enhances glucose transport of L6 myotubes in an insulin-independent pathway in a time- and dose-dependent manner.

A Case of Bacteremia by Atopobium rimae in a Patient with Liver Cirrhosis / 대한진단검사의학회지

Atopobium rimae, previously Lactobacillus rimae, is a strictly anaerobic, non-spore forming grampositive rod which was frequently isolated from odontogenic infection. We report a case of A. rimae bacteremia. A 47-yr-old man with liver cirrhosis was admitted to the hospital via emergency room due to fever and chill. His abdominal and pelvic computed tomography revealed a small abscess near the left adrenal gland. Three sets of blood cultures were taken and non-spore forming, grampositive rods were detected in all anaerobic vials. This isolate grew small nonhemolytic, gray-white translucent colonies on Brucella blood agar and was obligatory anaerobic on air-tolerance test. This organism was negative for catalase, indole, nitrate-reduction and beta-lactamase and failed to identify by Vitek ANI card (bioMerieux, France). 16S rRNA sequences of this showed 99.8% homology of the published sequence of A. rimae (GenBank accession number AF292371). Aspirates of periadrenal abscess grew Escherichia coli and Peptostreptococcus micros. He was treated with metronidazole and imipenem and follow-up cultures of blood were negative at days 4 and 10. To our knowledge, this is the first report of bacteremia of A. rimae.

Cloning and expression of Par6A cDNA / 基础医学与临床

Objective Cloning and expression of Par6A.Methods Par6A cDNA was amplified from rat L6 skeletal muscle cells by RT-PCR and the cloning and expression vectors of Par6A were constructed.The expression vector was transfected into 293 cells.Furthermore,the function of Par6A was confirmed by Co-immunoprecipitation.Results Par6A cDNA with approximately 1 kb in length was successfully amplified,and the expression vector of pDsRed-Express-N1-Par6A was constructed.The red fluorescene was seen under fluorescent microscope after 293ET cells were transfected for 24 h using the pDsRed-Express-N1-Par6A vector.The expressed Par6A protein can interacte with PKC?.Conclusion We successfully cloned the Par6A cDNA from rat L6 skeletal muscle cells,which provided a reliable method to study the function of Par6A.