ObjectiveTo observe the effect of Batroxobin and Urokinase on brain of diabetic rats following focal cerebral ischemia and reperfusion injury. To investigate the preventive mechanism of Batroxobin following focal cerebral ischemia and reperfusion injury in diabetic rats after thrombolysis therapy.MethodsDiabetic rat was induced by administrating streptozotocin intraperitoneally. Rat model of focal cerebral ischemia and reperfusion injury induced by intraluminal filament occlusion of middle cerebral artery(MCA) that removed 2h later was used. Batroxobin and Urokinase were administrated intravenously in different groups. Infarct volume,cerebral hemorrhage and matrix metalloproteinase (MMP)-2,MMP-9 were detected at 2h,24h,48h after ischemia and reperfusion injury.ResultsThe significant decrease of infarct volume were observed in Batroxobin and Urokinase groups. There were 5 rats observed cerebral hemorrhage in Urokinase group and no cerebral hemorrhage in Batroxobin group. The number of MMP-2 and MMP-9 positive cells in Batroxobin and Batroxobin Urokinase groups decreased compared with saline and Urokinase groups. ConclusionBatroxobin can decrease the infarct volume significantly without the complication of cerebral hemorrhage after ischemia and reperfusion injury in diabetic rats, which maybe relate to down regulation of the expression of MMP-2 and MMP-9.

AIMTo investigate whether free fatty acid impair insulin signaling proteins to inhibit insulin action in rat islet cells.

METHODSAfter rat islet cells were incubated with palmitate (0.25 mmol/L) or oleate (0.125 mmol/L) for 12 hours, 24 hours and 36 hours, western bolt was used to assess the protein abundance of cPKCalpha, Grb2 and FERK2.

RESULTSThe protein content of cPKCalpha were significantly upregulated and the protein abundance of Grb2 and ERK2 was decreased comparing with control in rat islet cells after incubated with free fatty acids.

CONCLUSIONSFree fatty acids may inhibit insulin activity in rat islet cells through up-regulating the expression of cPKCalpha or down-regulating the expression of Grb2 and ERK2.

Animals ; Cells, Cultured ; Fatty Acids, Nonesterified ; blood ; GRB2 Adaptor Protein ; metabolism ; Insulin ; metabolism ; Islets of Langerhans ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Protein Kinase C-alpha ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Objective To explore the clinicopathological characteristics of early gastric cancer (EGC),surgical treatment, and prognosis in elderly patients. Methods The clinicopathological and follow-up data in 83 elderly patients with EGC treated by gastrectomy from 1989. 8-2004. 8 were studied retrospectively. Results All of 83 patients were resectable, and there was no operative death nor surgical complications. The 5-, 10-, and 15- year survival rate was 98%,95%,and 92%, respectively. Sixty-one cases (74%) were mucosal cancer without lymph node metastasis, 22 cases were submucosal cancer with node metastasis in 4 cases. 7 cases were multiple primary carcinoma. Conclusions Local resection and partial gastrectomy are not suitable for multiple primary carcionoma in elderly patieats with EGC. Subtotal gastrectomy, and total gastrectomy should be the procedure of choice when foci were scattered about. Lymph node metastasis,multiple primary cancer foci and invasion of submucosal layer are all among factors impacting postoperative prognosis.

Aged ; Aged, 80 and over ; China ; epidemiology ; Dengue ; epidemiology ; Female ; Humans ; Male ; Middle Aged

Objective To explore the influence of hypoxia on cystathionine gamma lyase (CSE) gene expr ession of rat cultured pulmonary artery smooth muscle cells (PASMCs) and the mod ulatory effect of hydrogen sulfide on PASMCs proliferation.Methods Rat′s cultured PASMCs were randomly divided into contr ol group,NaHS group,hypoxia group and hypoxia +NaHS group. Each group contains 6 wells of cells. Hypoxic challenge was performed with oxygen concentr ation≤1%. The cell proliferation rate was measured by 3H-thymidine i ncorporation into DNA of the PASMCs. The expression of CSE mRNA was measured by quantitative competitive reverse-transcription polymerase chain reaction (RT-PCR ).Results For the PASMCs in hypoxia group, the amount of CSE mRN A was down-regulated about 67% (P

OBJECTIVETo explore the mechanism underlying the treatment of rheumatoid arthritis by Keshiling (KSL) in the rats model of FCA-induced arthritis.

METHODThe experimental arthritis was induced by FCA in the rats. The content of PGE2 in the inflammatory swelling toes was evaluated by ultraviolet spectrophotometric method. The ConA and LPS-induced lymphocytes proliferation and the production of interleukin-2 (IL-2) secreted by thymus were determined by MTT assay.

RESULTResults showed that the increases of lymphocyte proliferation and IL-2 production in AIA rats could be inhibited by KSL at the concentrations of 540 and 270 mg x kg(-1) in vivo and vitro. KSL at the same doses decreased the contents of PGE2 in inflammatory swelling toes, and the decreased A values were 25.6,16.1, 10.0 (A x 10(3)), respectively. After administration of KSL in vivo at 540 and 270 mg x kg(-1) the T lymphocyte proliferation were attenuated by 32.1% and 31.0%, and the production of IL-2 was inhibited by 17.5% and 14.0% respectively. While the inhibitory rates of T lymphocyte proliferation were reduced by 39.0% and 22.1% and the production of IL-2 was diminished by 27.3% and 18.2% respectively following the administration of KSL in vitro.

CONCLUSIONKSL possesses the anti-inflammation function.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; pharmacology ; Ants ; chemistry ; Arthritis, Experimental ; metabolism ; pathology ; Cell Proliferation ; Dinoprostone ; metabolism ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Interleukin-2 ; metabolism ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; metabolism ; pathology

Xiaoyao pills are a famous traditional Chinese medicine collected in Welfare Pharmacy, which is a classic prescription for treating liver depression and spleen deficiency. However, its composition is complex. In order to better control the quality of Xiaoyao pills, in this study, HPLC-ion-trap time-of-flight mass spectrometry (LC-IT-TOF/MS) was used to identify the main ingredients of Xiaoyao pills, paeoniflorin, albiflorin, glycyrrhizic acid, saikosaponin A and saikosaponin B2. Then a liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed for simultaneous determination and quantification of the main compounds. Fragmentation pathways of five active components were obtained. The method was validated. Five active ingredients in Xiaoyao pills had a good linear relationship, and the values of RSD (%) of repeatability were all less than 5%, the recovery ranges were between 90% and 115%, and the values of RSD (%) of each substance were less than 10% after the sample solution is placed for 24 hours. Three batches of Xiaoyao pills (concentrated pellets) and two batches of Xiaoyao pills (water pellets) were determined, the contents of paeoniflorin in concentrated pills were more than 4.0 mg·g^-1, and those in water pills were more than 2.5 mg·g^-1, which was accordance with Chinese Pharmacopoeia. However, other compounds behave differently. This method has high sensitivity and reliable measurement results, which provides basis for quality control of Xiaoyao pills and material basis for pharmacology research.

Objective:To develop a rapid and quantitative detection method for abrin using colloid gold immunochromatographic assay. Methods:The rapid detection method was established with double-antibody sandwich assay. Quantification was realized by constructing a standard curve. The specificity and sensitivity of the method were tested, and its feasibility was evaluated by various abrin-added food samples. Results and Conclusion:This established method could accomplish qualitative and semi-quantitative detection in 15 minutes; the sensitivity was up to 30 ng/ml with a linear range from 30 ng/ml to 600 ng/ml. The recovery rate was 80%-110%, and the variation coefficient was less than 15%.The colloidal gold immunochromatographic assay is rapid, specific, sensitive,accurate and suitable for field detection.

This study was purposed to investigate the efficacy of different whole flow lysis reagents for lysis of red blood cells in flow cytometric analysis. The expression of immunocytes was detected by flow cytometry after lysis of red blood cells using commercial reagents (Optilyse C, FACS Lysing Solution) and self-made red blood cell lysis reagents (RBC Lysis Buffer), the detection results were analyzed comparatively. The results showed that there was no significant difference in the percentage of CD3e(+), CD3e(+)CD4(+), CD3e(+)CD8a(+), CD3e(-)CD19(+), CD3e(-)NK1.1(+) and Gr-1(+) cells between 3 different lysis reagent groups. However OptiLyse C solution was suitable to Gr-1(+) cell detection, but did not suit to Foxp3(+) Treg detection. The self-made RBC Lysis Buffer and FACS Lysing Solution were suited to Foxp3(+) Treg detection. It is concluded that the use of self-made RBC Lysis Buffer for flow cytometry can get the lysis efficiency of commercially available lysis solutions when samples are prepared in accordance with standardized procedure. The self-made RBC Lysis Buffer not only can satisfy experimental requirements, but also can reduce the experimental costs.
Animals ; Erythrocyte Count ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; instrumentation ; methods ; Immune System ; immunology ; Indicators and Reagents ; analysis ; Mice ; Mice, Inbred C57BL

·AIM:To determine the effect of anterior-posterior joint surgery on choroidal thickness in patients with proliferative diabetic retinopathy (PDR). · METHODS: A retrospective, case - control study enrolled 60 eyes of 60 patients with PDR diagnosed at Qingdao Municipal Hospital. The patients, who had conditions that warranted anterior - posterior joint surgery,were divided into a clinically significant macular edema group (PDR/CSME+;31 patients,31 eyes) and a non-CSME group (PDR/CSME-;29 patients,29 eyes). Twenty-seven eyes of 27 normal patients were included in the control group. All affected eyes underwent anterior - posterior joint surgery. After surgery, the subfoveal choroidal thickness (SFCT), and the nasal choroidal thickness (NCT) and temporal choroidal thickness (TCT), which were obtained at a distance of 1500μ m from the fovea in the nasal and temporal directions, respectively, were measured in the control and PDR groups by enhanced depth imaging spectral domain optical coherence tomography (EDI-SDOCT) at 1wk,1,3, and 6mo after surgery. Changes in choroidal thickness after anterior - posterior joint surgery were compared between the groups. ·RESULTS:The SFCT,NCT,and TCT were significantly thicker at 1mo than at 1wk, 3, and 6mo after surgery in the PDR/CSME+ and PDR/CSME- groups(P<0.05). The SFCT, NCT, and TCT were significantly thinner at 6mo than at 1wk,1,and 3mo after surgery in the PDR/CSME+and PDR/CSME- groups(P<0.05). The SFCT,NCT,and TCT in the PDR/CSME+ and PDR/CSME- groups at 1wk, 1, and 3mo after surgery were significantly thicker than those in the control group (all P<0.05), but the SFCT, NCT, and TCT at 6mo after surgery showed no significant difference compared with the control group (all P>0.05). There was no significant difference in the SFCT,NCT, or TCT at 1wk, 1, 3, or 6mo between the PDR/CSME+ and PDR/CSME- groups (P>0.05). ·CONCLUSION:The choroidal thickness of PDR patients increases within 1mo after surgery, and decreased after 1mo,but is not significantly different between the control group and the PDR groups at 6mo after surgery. Whether PDR is associated with CSME has no effect on the choroidal thickness after surgery.