Objective To investigate the effect of insulin and gliclazide therapy on endoplasmic reticulum (ER) stress and insulin sensitivity in the liver of type 2 diabetic rats.Methods A high fat diet plus a low-dose of streptozotocin was implemented to create a type 2 diabetic rats which were randomly divided into diabetes mellitus (DM) group,insulin treatment (INS) group and gliclazide treatment (GT)group; and healthy rats were as normal control group.Diabetic rats in INS and GT groups were given neutral protamine hagedorn (NPH) insulin and gliclazide respectively for 3 weeks.Protein expression levels of immunoglobulin binding protein (Bip),spliced X-box binding protein 1 (XBP-ls),phosphorylated c-Jun on serine 73 (p-c-Jun),phosphorylated insulin receptor substrate 1 on serine 307 (p-IRS-1),and glucose-6-phosphatase (G6Pase) in liver homogenate were detected by Western blotting.Results Compared with the normal rats,Bip and XBP-Is in the DM group were up-regulated (0.28 ±0.07 vs 0.90 ±0.10 for Bip;0.41 ± 0.07 vs 0.95 ±0.07 for XBP-1 s; both P ＜ 0.01 ) ; p-c-Jun (0.59 ± 0.18 vs 1.94 ± 0.03 ),p-IRS-1( 1.73 ± 0.18 vs 5.32 ± 0.22) and G6Pase (0.11 ± 0.01 vs 0.45 ± 0.01 ) were increased ( all P values ＜0.01 ).In the INS group,all of aforementioned changes were reversed (0.90 ± 0.10 vs 0.25 ± 0.04 for Bip; 0.95 ±0.07 vs 0.47 ±0.01 for XBP-1s; 1.94 ± 0.03 vs 0.50 ±0.10 for p-c-Jun; 5.32 ± 0.22 vs 1.59 ±0.32 for p-IRS-1 ; 0.45 ±0.01 vs 0.15 ±0.02 for G6Pase,all P values ＜0.01 ).In the GT group,all of aforementioned changes were also attenuated ( 0.90 ± 0.10 vs 0.53 ± 00.02 for Bip ; 0.95 ± 0.07 vs 0.78±0.02 for XBP-1s; 1.94 ±0.03 vs 1.33 ±0.11 for p-c-Jun; 5.32 ±0.22 vs 3.13 ±0.02 for p-IRS-1; 0.45 ± 0.01 vs 0.25 ± 0.01 for G6Pase,all P values ＜ 0.05).Furthermore,all of aforementioned protein levels were down-regulated more obviously in the INS group comparing to the GT group ( all P values ＜ 0.01 ).Conclusions Both insulin and gliclazide therapy could relieve ER stress and e-Jun N-terminal kinase activity and improved insulin sensitivity.The effect of insulin on Bip,XBP-1s,p-c-Jun,p-IRS-1 and G6Pase protein expressions is more obvious than that of glilcazide,which indicates besides lowering glucose,insulin might have protective effects of anti-inflammation,anti-oxidative stress or stimulation of lipid redistribution.

The effect of insulin treatment on fatty liver was investigated in diabetic mice fed with high-fat diet.C57 BL/6J mice were fed with high fat diet for 12 weeks,and then treated with Glargine for 4 weeks.The results showed that during intraperitoneal glucose tolerance test,blood glucose,serum total cholesterol and triglyceride(TG) were significantly lower in insulin-treated high fat group than those in untreated ones(P<0.05).The hepatic histology showed minimal or barely visible fat in hepmic cells along with TG content in liver decreasing by 20.71%. The results suggest that insulin ameliorates hepatic intracellular lipid accumulation induced by high fat diet.

[Objective]To explore the intracellular antimicrobial activities of erythromycin,ciprofloxacin,levofloxacin,moxifloxacin against Legionella pneumophila.[Methods]The minimum inhibition concentration(MIC)of each antibiotic was evaluated by E-test method and microdilution method respectively.The minimal extracellular concentration inhibiting intracellular multiplication(MIEC)of each antibiotic was evaluated by the MTT colorimetric assay system.[Results]The MIC concentration for each drug by E-test method were:erythromycin,0.047 μg/mL;ciprofloxacin,0.38 μg/mL;levofloxacin,0.125 μg/mL;moxifloxacin,0.125 μg/mL,the MIC concentrations for each drug by microdilution method were:erythromycin,0.125 μg/mL;ciprofloxacin,0.03 μg/mL;levofloxacin,0.016 μg/mL;moxifloxacin,0.016 μg/mL.The MIEC concentration for each drug were:erythromycin,0.25 μg/mL;ciprofloxacin,0.016 μg/mL;levofloxacin,0.016 μg/mL;moxifloxacin,0.004 μg/mL.[Conclusions]Fluoroquinolones have superior activity than erythromycin in U937 cells infected with L.pneumophila.Moxifloxacin is the most potent drug among the four tested antimicrobials.Our results indicated that the MTT assay system allows comparative and quantitative evaluations of the intracellular activities of antibiotics against L,pneumophila and efficient processing of a large number of samples.

OBJECTIVETo identify triacylglycerols in coix oil.

METHODHigh performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry was used for identification. The experiment was operated under the conditions: spray voltage at 3 000 V, capillary temperature at 250 degrees C, APCI vaporizer temperature at 400 degrees C, and corona current of 4 microA. Sheath gas pressure (high purity liquid nitrogen) was 35 kPa. Mass spectra were obtained over the m/e range of 300 to 900 amu, scan duration of 1s and Q1 peak width at 0.7. The stationary phase was Zorbax Extend C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase: dichloromethane-acetonitrile (35:65), flow rate: 1 mL x min(-1); column temperature: 25 degrees C.

RESULT12 triacylglycerols were identified by HPLC-MS method.

CONCLUSIONThe result can be used to identify the components in a fingerprint chromatogram of coix oil and its related injection product.

Chromatography, High Pressure Liquid ; methods ; Coix ; chemistry ; Mass Spectrometry ; methods ; Plant Oils ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Seeds ; chemistry ; Triglycerides ; analysis ; chemistry ; isolation & purification

Objective To investigate the effect of early insulin therapy on NF-KB pathway and inflammatory cytokine responses in fiver of diabetic rat.Methods NF-KB p65 DNA binding was assayed with ELISA-based assay kit,cytokine gene expressions were quantified with real-time PCR and phosphoenolpyruvate carboxykinase(PEPCK),NF-KB and inhibitor KB(IKBα)protein levels wlere assayed with Westem blot.Results Compared with control,hepatic PEPCK protein level in the untreated diabetic rat increased by 40%.Early insulin and gliclazide treatment normalized PEPCK protein level.The abundance of IKBα protein was significantly decreased and nuclear NF-KB p65 DNA binding activity was incteased in untreated diabetic rats.IKBot protein content increased and NF-KB p65 DNA binding decreased during early intervention treatment.mRNAs encoding IL-1β and TNFα were increased,which were reduced to normal levels after insulin and gliclazide treatment.Conclusions It is suggested that early insulin treatment inhibits NF-KB activity and inflammatory cytokine responses in fiver that are involved in the aniefioration of insulin resistance in diabetic rats.Such results misht be due to indirect antiinflammatory effects of insulin thus relieving glucotoxicity and lipotoxicity in pefipherM tissues.

Objective To enhance the management in identifying the surgical sites to comply with national standards.Methods A nurse-physician collaboration management team was set up to investigate the current identification of surgical sites in every operating room which violates regulations,with the causes analyzed and countermeasures proposed.Working hand in hand,doctors and nurses figured out the management details for preoperatively identifying the surgical sites and reengineering of the surgical process.With the responsibilities clarified and training enhanced,the surgeons,anesthesiologists,ward nurses and operating room nurses were held responsible for the process and improvements of identifying the surgical sites.The number of patients with unreasonable identification of surgical sites was calculated before and after establishment of nurse-physician collaboration management team.Results The reasonable identification rates of surgical sites were 37.94% before the reform and 80.94% after;incorrect use of all types of the surgical site identification can be minimized in the reform.Conclusion The management of nurse-physician collaboration is conducive to enhancing the reasonable identification rate of surgical sites,thus improving the quality of care and correctness of operations.

To study the preventive effect of sophocarpine (Soc) on dextran sulfate sodium (DSS)-induced colitis in mice, in order to analyze the influence of Soc on toll like receptor 4 (TLR4)/mitogen-activated protein kinases (MAPKs) and janus tyrosine kinase 2 signal transducer and activator of transcription 3 (JAK2/STAT3) signal pathways in mice intestinal tissues. The mice was given 2.5% DSS for 6 days to induce the acute colitis model. The Soc-treated group was intraperitoneally injected with sophocarpine 30 mg · kg(-1) · d(-1) since the day before the experiment to the end. The disease activity index (DAI) was assessed everyday, and the colonic morphology and histological damage were observed with HE staining. The mRNA expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were detected by real-time RT-PCR. The changes in key protein kinase p38 mitogen-activated protein kinase (p38MAPK), c-Jun NH2-terminal protein kinase1/2 (JNK1/2), extracellular signal-regulated kinase1/2 (ERK1/2), JAK2, STAT3 in TLR4/MAPKs and JAK2/STAT3 signaling pathways were detected by western blot. The result showed that the model group showed statistical significance in body weight, DAI, colon length and histopathological changes compared with the normal group (P <0.05); however, the Soc-treated group showed significant improvements in the above indexes compared with the model group (P <0.05). TNF-α, IL-1β and IL-6 in the model group was significantly higher than that in the normal group (P <0.05), but lowered in the Soc-treated group to varying degrees (P <0.05). In the normal group, the expressions of TLR4 and the phosphorylation of P38, JNK1/2, JAK2, STAT3 were at low levels; in the model group, the phosphorylation of P38, JNK1/2, JAK2, STAT3 increased; the Soc-treated group showed a decrease in TLR4 expression compared with the model group, with notable declines in the phosphorylation of TLR4, P38, JNK1/2, JAK2, STAT3. These findings indicate that Soc can inhibit TLR4/MAPKs, K2/STAT3 signaling pathway activation, reduce the expression of proinflammatory cytokines TNF-α, IL-1β and IL-6 and relieve inflammatory reactions, so as to effectively prevent experimental colitis.
Alkaloids ; pharmacology ; therapeutic use ; Animals ; Colitis ; drug therapy ; immunology ; pathology ; Cytokines ; genetics ; Janus Kinase 2 ; antagonists & inhibitors ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Phosphorylation ; STAT3 Transcription Factor ; antagonists & inhibitors ; physiology ; Toll-Like Receptor 4 ; antagonists & inhibitors ; physiology

OBJECTIVETo explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats.

METHODSSixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated.

RESULTSDutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05).

CONCLUSIONDutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and β-catenin expressions, and damaging epididymal epithelial cell junctions.

Animals ; Azasteroids ; pharmacology ; Claudin-1 ; metabolism ; Dihydrotestosterone ; blood ; Dutasteride ; Epididymis ; drug effects ; metabolism ; Female ; Fertility ; drug effects ; Humans ; Intercellular Junctions ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Testosterone ; blood ; Urological Agents ; pharmacology ; beta Catenin ; metabolism

BACKGROUNDAdipocytes behave like a rich source of pro-inflammatory cytokines including monocyte chemoattractant protein-1 (MCP-1). Oxidized low-density lipoprotein (oxLDL) participates in the local chronic inflammatory response, and high-density lipoprotein could counterbalance the proinflammatory function of oxLDL, but the underlying mechanism is not completely understood. This study aimed to evaluate the effect of apolipoprotein A-I mimetic peptide L-4F on the secretion and expression of MCP-1 in fully differentiated 3T3-L1 adipocytes induced by oxLDL and to elucidate the possible mechanisms.

METHODSFully differentiated 3T3-L1 adipocytes were incubated in the medium containing various concentration of L-4F (0-50 μg/ml) with oxLDL (50 μg/ml) stimulated, with/without protein kinase A (PKA) inhibitor H-89 (10 μmol/L) preincubated. The concentrations of MCP-1 in the supernatant, the mRNA expression of MCP-1, the levels of CCAAT/enhancer binding protein α (C/EBPα), and CCAAT/enhancer binding protein β (C/EBPβ) were evaluated. The monocyte chemotaxis assay was performed by micropore filter method using a modified Boyden chamber.

RESULTSOxLDL stimulation induced a significant increase of MCP-1 expression and secretion in 3T3-L1 adipocytes, which were inhibited by L-4F preincubation in a dose-dependent manner. PKA inhibitor H-89 markedly reduced the oxLDL-induced MCP-1 expression, but no further decrease was observed when H-89 was used in combination with L-4F (50 μg/ml) (P > 0.05). OxLDL stimulation showed no significant effect on C/EBPα protein level but increased C/EBPβ protein level in a time-dependent manner. H-89 and L-4F both attenuated C/EBPβ protein level in oxLDL-induced 3T3-L1 adipocytes.

CONCLUSIONSOxLDL induces C/EBPβ protein synthesis in a time-dependent manner and enhances MCP-1 secretion and expression in 3T3-L1 adipocytes. L-4F dose-dependently counterbalances the pro-inflammatory effect of oxLDL, and cyclic AMP/PKA-C/EBPβ signaling pathway may participate in it.

3T3-L1 Cells ; Animals ; CCAAT-Enhancer-Binding Protein-beta ; analysis ; physiology ; Chemokine CCL2 ; genetics ; secretion ; Cyclic AMP ; physiology ; Cyclic AMP-Dependent Protein Kinases ; physiology ; Humans ; Lipoproteins, LDL ; antagonists & inhibitors ; pharmacology ; Mice ; Peptides ; pharmacology ; Signal Transduction ; physiology

OBJECTIVEStudy the characteristics of absorption and separation of traditional Chinese medicine compound prescription using macroporous resin.

METHODStudy the techniquecs and characteristics of absorption and separation of a sample by macroporous resin, which is composed of coptis root, rhubarb and common anemarrhena rhizome, containing alkaloid, anthraquinone and saponin.

RESULTIt is proved by qualitative and quantitative researches studies that after absorbed and separated by optimized technics process, most prime effective components or section fractions in traditional Chinese medicine compound prescription can be reserved maintained.

CONCLUSIONIf the techniquecs of separation is properly designed, the same kind of macropore resin can absorbd and separate various effective components or section in traditional Chinese medicine compound prescription which have with different chemical structures efficiently.

Alkaloids ; isolation & purification ; Anemarrhena ; chemistry ; Anthraquinones ; isolation & purification ; Coptis ; chemistry ; Plants, Medicinal ; chemistry ; Resins, Synthetic ; Rheum ; chemistry ; Saponins ; isolation & purification ; Technology, Pharmaceutical ; methods