Objective To explore the role of stromal interaction molecule 1(STIMI)on prohteration and intra- cellular Ca~(2+)change in vascular smooth muscle ceils(VSMC).Methods Rat VSMC were isolated from SD rats and primary cultured.Ad-si/rSTIM1 and Ad-hSTIM1 were transfected into VSMC.The protein of STIM1 was measured by Western blot,the proliferation of VSMC was analyzed by ~3H-thymidine(~3H-TdR)incorporation and cell count,the intracellular Ca~(2+)change was assessed By Aquaeosmos system.Ruselts Fourty-eight hours after transfection,as compared with Ad-hSTIMI group,the Ad-si/rSTIMI VSMC had lower expression of STIM1 protein (P

Objective To study the effect of bromocriptine(BRC) on the activation of T lymphocyte stimulated by phytohemagglutinin(PHA).Methods After CD4+ T cell line Jurkat E6-1 cells were stimulated by PHA,prolactin(PRL) and BRC,respectively,the expression of linker for activation of T cells(LAT) and zeta-chain T cell receptor associated protein kinase 70 000(ZAP-70) mRNA of T lymphocytes were checked by RT-PCR.The expression of PRL mRNA of T lymphocytes was detected by Real time PCR.The expression of CD25(cluster of differentiation) as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry,and the activation of nuclear factor-?B(NF-?B) was detected by luciferase reporter system.Results 1.BRC inhibited the expression of ZAP-70 as the common signal molecules both in the T lymphocyte activation pathway and PRL-prolactin-prolactin receptor(PRLR) signal transduction pathway,and decreased the expression of PRL mRNA produced by activation T lymphocytes.2.BRC enhanced the expression of LAT mRNA as another important signal molecular on the T lymphocytes and CD25 on the surface of the T lymphocytes.3.The activation of NF-?B of T lymphocytes was decreased.Conclusions BRC might inhibit the activation of T lymphocytes by inhibiting the expression of ZAP-70,the common signal molecules between T lymphocytes activation and PRL-PRL pathway,and PRL mRNA,the like-T lymphocyte growth factor.

OBJECTIVETo evaluate the efficacy and safety of balloon dilation (BD) with gastroscope in treatment of esophageal stricture in children.

METHODSBD was performed in 12 children aged 5 - 59 months, average age 26 months, course of disease was 2 - 26 months, with esophageal stricture, 7 cases with anastomotic strictures secondary to surgical repair of esophageal atresia, 3 with congenital esophageal stenosis, 2 with corrosive esophageal strictures. All procedures were performed under tracheal intubation and intravenous anesthesia using the 3rd grade controlled radial expansion (CRE) esophagus-balloon with gastroscope. Firstly the balloon was inserted into the esophagus through mouth, then put in the gastroscope. Under the direct guidance of gastroscope the balloon was positioned across the stricture, then the balloon was filled with saline to get needed pressure and maintained for 3 minutes. The procedure was repeated 3 times at an interval of 3 minutes. The abdominal pain, melena and vomiting were observed, as well as the diet taken thereafter, the size of the stricture and the nutrition status were observed for 3 to 12 months after the dilation.

RESULTSTwenty-two dilations were performed in 12 cases, 19 succeeded, 3 cases developed complication during the dilation, the total success rate was 86%. The procedure failed in 3 cases and succeeded in 9 cases, the effective rate was 75%. Follow-up and repeated gastroscopy were performed within 3 to 12 months after the dilation, the diameter of the stricture was 9-13 mm, compared with 2-8 mm before the dilation. Eight of the children could take solid food and nutritional status was improved.

CONCLUSIONSBD with the 3rd grade CRE esophagus-balloon under gastroscopy is a simple and effective method to treat esophagus stricture in children, especially for anastomotic strictures secondary to surgical repair of esophageal atresia.

Catheterization ; methods ; Child, Preschool ; Esophageal Stenosis ; therapy ; Gastroscopes ; Humans ; Infant ; Treatment Outcome

OBJECTIVETo investigate the role of store-operated calcium channels (SOCs) in primary hepatocytes under conditions of calcium overload and ethanol-induced injury.

METHODSThe in vitro model of chronic ethanol-induced hepatocyte injury was established using primary hepatocytes isolated from Sprague-Dawley rats. Ethanol-induced changes (24, 48 and 72 h; 50, 100, 200, 400 and 800 mmol/L) in expression of the SOCs proteins stromal interaction molecule 1 (STIM1) and calcium release-activated calcium channel protein 1 (Oria1) were detected by qualitative PCR analysis (mRNA) and western blotting (protein). The possible role of these two SOCs proteins in the ethanol-induced extracellular calcium influx and related liver cell injury was determined by treating the cell system with various channel blockers (EGTA, La3+, and 2-APB). Cell viability was determined by MTT assay and cytosolic free calcium ion concentration was determined by flow cytometry.

RESULTSAfter 24 h of exposure to 0 (untreated) to 800 mM/L ethanol, the cell viability was reduced in a concentration-dependent manner. The 400 mmol/L concentration of ethanol decreased cell viability by 57.34% +/- 2.34%. and was chosen for use in subsequent experiments. Compared with the untreated control cells, the ethanol-treated cells showed significantly up-regulated mRNA and protein expression of both STIM1 and Orai1 at all times examined, suggesting that the ethanol-stimulated expression of STIM1 and Orai1 could persist for at least 72 h. The ethanol treatment induced increase in cytoplasmic calcium levels was significantly (and similarly) reduced by co-treatment with any of the three channel blockers.

CONCLUSIONChronic ethanol exposure can increase the expression of STIM1 and Orai1 in primary liver cells, suggesting that ethanol may increase extracellular calcium influx by up-regulating expression of these SOCs protein molecules, ultimately aggravating liver cell damage.

Animals ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels ; metabolism ; Calcium-Binding Proteins ; metabolism ; Cell Survival ; Cells, Cultured ; Ethanol ; adverse effects ; Hepatocytes ; drug effects ; metabolism ; Male ; Membrane Glycoproteins ; metabolism ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Stromal Interaction Molecule 1

OBJECTIVETo study the expression of Toll-like receptor 4 (TLR4) in renal tubular epithelial cells exposed to high glucose and the effect of spironolactone on the TLR4 expression.

METHODSIn vitro renal tubular epithelial cells (NRK-52E) were randomly exposed to DMEM culture solution with low glucose (5 mmol /L), high glucose (25 mmol/L) or 10(-7) mol/L spironolactone plus 25 mmol/L glucose. Immunohistochemistry, RT-PCR and Western blot were used to determine TLR4 protein and mRNA expression. The levels of IL-6 and TNF-alpha in the cell culture supernatant were determined using ELISA.

RESULTSThe expression of TLR4 mRNA in the high glucose group began to increase 6 hrs and remained at a higher level up to 24 hrs after exposure as compared with the low glucose group. The TLR4 mRNA expression in the spironolactone treatment group was significantly lower than that in the high glucose group, although it was higher than that in the low glucose group between 6 and 24 hrs after exposure. TLR4 protein expression increased significantly in the high glucose group 24 and 48 hrs after exposure compared with that in the low glucose group. The TLR4 protein expression in the spironolactone treatment group was lower than that in the high glucose group, but higher than that in the low glucose group. IL-6 and TNF-alpha expression in the supernatant from the NRK-52E cells in the high glucose groups increased significantly as compared with the low glucose group. The spironolactone treatment group had significantly reduced IL-6 and TNF-alpha expression compared with the high glucose group.

CONCLUSIONSHigh glucose triggers an increase in the expression of TLR4 and inflammatory factors in NRK-52E cells. TLR4 may participate in the progress of diabetic nephropathy. Spironolactone can reduce expression of TLR4 and inflammatory factors, which might be attributed to one of the mechanisms of protection by spironolactone against diabetic nephropathy.

Cells, Cultured ; Diabetic Nephropathies ; etiology ; Epithelial Cells ; metabolism ; Humans ; Hyperglycemia ; metabolism ; Immunohistochemistry ; Interleukin-6 ; analysis ; Kidney Tubules ; metabolism ; RNA, Messenger ; analysis ; Spironolactone ; pharmacology ; Toll-Like Receptor 4 ; analysis ; genetics ; physiology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo explore the clinical effect and possible mechanism of Shengxueling (SXL), a Chinese medical preparation mainly consisting of ginseng saponins, in treating refractory idiopathic thrombocytopenic purpura (ITP).

METHODSThe selected 69 patients with ITP were randomly assigned to two groups, the 37 patients in the treated group were treated orally by SXL with the dose for adult as 60 mg twice a day for two weeks. Then when no marked rise of platelet count after that, the dose would be doubled and administered for another two weeks. Then the dose could be gradually reduced to the initiative level in patients who responded to the treatment, and if they did not, the treatment was regarded as ineffective and be terminated. The 32 patients in the control group were treated with ampeptide elemente instead of SXL, 0.4 g each time three times a day in the first two weeks, and, if that was ineffective, 0.2 g would be added each time and 1.8 g would be administered a day for two more weeks. Four weeks' treatment was regarded as one therapeutic course for both groups and the observation lasted for two successive courses in patients showing positive reslonse.

RESULTSIn the 37 patients in the treated group, markedly effective was obtained in 7 (19.0%), favorably effective in 15 (40.5%), improved in 5 (13.5%) and ineffective in 10 (27.0%), the total effective rate being 59.5%. The corresponding number in the 32 patients in the control group was 4 (12.5%), 6 (18.8%), 3 (9.4%), 19 (59.4%) and 31.3% respectively. Comparison showed the difference in therapeutic efficacy between the two groups was significant (P<0.05).

CONCLUSIONSXL is a safe and effective preparation for treatment of ITP, showing an immediate effect which is obviously superior to that of ampeptide elemente with less adverse effect.

Administration, Oral ; Adolescent ; Adult ; Amino Acids, Essential ; therapeutic use ; Bone Marrow ; pathology ; physiopathology ; Child ; Drug Administration Schedule ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Megakaryocytes ; pathology ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; blood ; physiopathology ; therapy ; Treatment Outcome

OBJECTIVETo study the predominant calcium-antagonist components of Danshen injection.

METHODThe effects of danshensu, protocatechualdehyde and Danshen injection on calcium concentration in cytoplasm of erythrocytes were examined in vitro by the fluorescent Ca+ -chelator fura-2.

RESULTEither DS182 or PCAD can decrease in dose-dependent cytosolic free calcium concentration in human erythrocytes. They had additive effect when mixed, which was similar to Danshen injection.

CONCLUSIONDS182 and PCAD may be predominant calcium-antagonist components of Danshen injection.

Adult ; Benzaldehydes ; isolation & purification ; pharmacology ; Calcium ; metabolism ; Catechols ; isolation & purification ; pharmacology ; Cytoplasm ; metabolism ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; metabolism ; Female ; Humans ; Injections ; Lactates ; isolation & purification ; pharmacology ; Male ; Middle Aged ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective:To observe the clinical effect of Kangfuxin solution combined with Almagate suspension on the complication of gastroscope biopsy. Method:Totally 276 cases of chronic superficial gastritis gastroscope biopsy were divided into treatment group and control group randomly. Treatment group was treated by Kangfuxin solution combined with Almagate suspension，while the control group was given no precautionary measures as usual. Then the patients received the abdominal symptom score and fecal occult blood test（FOBT）after 1 week. Result:The cases with abdominal pain，burning sensation and stool occult blood in the control group were more than the treatment group significantly（P<0.05）. The patients' abdominal symptom score in control group was higher than that before the gastroscope biopsy，and that was lower in treatment group（P<0.05）. And the patients' abdominal symptom score of the control group was significantly higher than that of the treatment group after the gastroscope biopsy（P<0.05）. Compared with the Helicobacter pylori（HP）-infected subgroup，the incidences of early satiety and ventosity in the control group were higher than those of the treatment group（P<0.05）. The abdominal symptom score of the patients infected with helicobacter pylori（HP）in control group was significantly higher than that before the gastroscope biopsy and in the treatment group（P<0.05）. Conclusion:Kangfuxin solution combined with Almagate suspension is safe and effective in preventing complications of gastroscope biopsy，especially for patients infected with HP. The method could avoid exacerbating clinical symptoms.