Objective To explore the effect of stroma cell-derived factor receptor CXCR4 on the homing of the hematopoietic stem/progenitor cells in utero transplantation. Methods CD34~+ cells were collected by Ficoil density gradient centrifugation and MiniMACS and then stimulated for 48 h by SCF and IL-6 cytokines prior to transplantation. CD184(CXCR4) expressions and transmigrate rates of the CD34~+ cells were analysed by flow cytometer. Cells pre-treated with different treatment were transplanted into the abdominal cavity of the fetal BALB/c mouse in the pregnant days 13～14. Human CD45 cells as the marker of graft were detected by flow cytometry after 1 month the fetus born. Results Expression changes of CD184 on CD34~+ cells were from 9. 58%±1. 56% to 19. 32%±3. 64% after SCF and IL-6 stimulation. The CD34~+/CXCR4~(high) cells exhibited significant increases in SDF-1 mediated chemotaxis compared with the CD34~+/CXCR4~(low) cells. Transmigration of CD34~+ /CXCR4~(high) was inhibited by pretreatment with an-tiCXCR4mAb and PTX. The positive rates of human CD45 cells detected in the fetal mouse were significantly higher in the SCF and IL-6 pretreatment group. This effects were significantly abrogated after the addition of antiCXCR4mAb or PTX. Conclusion Up-regulation of CXCR4 expression may be useful for improving hematopoietic stem/progenitor cells homing in utero transplantation. This homing process is mediated and depends on the CXCR4 receptors. The signal transduction is mediated by PTX-sensitive Gi protein.

Objective To explore the effect of stroma cell-derived factor receptor CXCR4 on the homing of the hematopoietic stem/progenitor cells in utero transplantation. Methods CD34+ cells were collected by Ficoll density gradient centrifugation and MiniMACS and then stimulated for 48 h by SCF and IL-6 cytokines prior to transplantation. CD184(CXCR4) expressions and transmigrate rates of the CD34+ cells were analysed by flow cytometer. Cells pretreated with different treatment were transplanted into the abdominal cavity of the fetal BALB/c mouse in the pregnant days 13～14. Human CD45 cells as the marker of graft were detected by flow cytometry after 1 month the fetus born. Results Expression changes of CD184 on CD34+ cells were from 9.58%?1.56% to 19.32%?3.64% after SCF and IL-6 stimulation. The CD34+/CXCR4high cells exhibited significant increases in SDF-1 mediated chemotaxis compared with the CD34+/CXCR4low cells. Transmigration of CD34+/CXCR4high was inhibited by pretreatment with antiCXCR4mAb and PTX. The positive rates of human CD45 cells detected in the fetal mouse were significantly higher in the SCF and IL-6 pretreatment group. This effects were significantly abrogated after the addition of antiCXCR4mAbor PTX. Conclusion Up-regulation of CXCR4 expression may be useful for improving hematopoietic stem/progenitor cells homing in utero transplantation. This homing process is mediated and depends on the CXCR4 receptors. The signal transduction is mediated by PTX-sensitive Gi protein.

OBJECTIVE:To study the protective effect of Weidean tablets on rats with stress-induced gastric ulceration. METH-ODS:Water immersion restraint stress method and empty stomach were employed to establish rat models with gastric ulceration. 60 SD rats were equally randomized into normal control group(isometric sodium chloride injection),model group(isometric sodium chloride injection),ranitidine group(0.015 g/kg),and groups of high,medium and low doses(1.70,0.87 and 0.43 g/kg)of Wei-dean tablets. General situation of stomach was observed .The gastric mucosal injury index,the NO content in serum,and the activi-ties of SOD and iNOS and the contents of MDA and PGE2 in the gastric tissue were detected for rats. Pathomorphological observa-tion of rats’gastric tissues was conducted under the microscope. RESULTS:Compared with normal control group,the rats in the model group had higher gastric mucosal injury index,lower content of NO in serum,and weaker activity of SOD,stronger activity of iNOS,higher content of MDA and PGE2,in the gastric tissue. There was statistical significance (P<0.01 or P<0.05). Dark matter,severe deformation and necrosis of gastric mucosal cells in rats were noted,as well as multiple large-area superficial ulcers. Compared with the model group,the rats in groups of high,medium and low doses of Weidean tablets had lower gastric mucosal injury index,higher content of NO in serum,and stronger activity of SOD,weaker activity of iNOS,lower content of MDA and higher content of PGE2,in the gastric tissue. There was statistical significance(P<0.01 or P<0.05). General situation of stomach and the pathomorphological condition of rats’gastric tissues were improved. CONCLUSIONS:Weidean tablets has protective ef-fect to some extent on rats with stress-induced gastric ulceration,the mechanism is related to the improvement of serum levels of anti-oxidation index in rats.

The aim of the current study is to investigate the effect of visfatin on cardiomyocyte hypertrophy. Cultured H9c2 cardiomyocytes were exposed to visfatin at different concentrations for different periods of time, and the markers of cardiomyocyte hypertrophy were detected. Moreover, pravastatin, the inhibitor of endoplasmic reticulum stress (ERS) or thapsigargin, an ERS agonist was used respectively to pre-treat the cells before visfatin stimulation. F-actin staining was performed to measure the cell surface change. The mRNA expressions of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and ERS markers including glucose-regulated protein 78(GRP78), C/EPB homologous protein (CHOP) and activating transcription factor 6 (ATF6) were assessed by real time RT-PCR. The change of protein level of GRP78 and CHOP was detected by Western blot. The experimental data demonstrated that exposure to 100 or 150 ng/mL concentrations of visfatin for 24 h, or 100 ng/mL of visfatin for 24 or 48 h, significantly increased the expression of markers for cardiomyocyte hypertrophy. Visfatin stimulation provoked ERS in H9c2 cells. Furthermore, pre-treatment with pravastatin partially inhibited the visfatin-induced mRNA expression of ANP and BNP in H9c2 cells, whereas thapsigargin promoted the visfatin-induced expression of cardiomyocyte hypertrophy markers. The results suggest that visfatin might induce cardiomyocyte hypertrophy via ERS -dependent pathways.
Actins ; Activating Transcription Factor 6 ; metabolism ; Animals ; Cell Line ; Heat-Shock Proteins ; metabolism ; Hypertrophy ; Myocytes, Cardiac ; cytology ; drug effects ; Natriuretic Peptide, Brain ; metabolism ; Nicotinamide Phosphoribosyltransferase ; pharmacology ; Rats ; Transcription Factor CHOP ; metabolism

ObjectiveTo investigate the electrocardiogram (ECG) characteristics of patients with early repolarization variant (ERV).Methods One hundred and six patients diagnosed of ERV based on clinical and ECG and the healthy control group of 100 patients were analyzed and ECG features and ERV site were compared between these two groups.Results The mean heart rate( [ 68.6 ± 8.4 ] beats/min vs [ 74.8 ± 12.6 ]beats/min),QRS time( [95 ± 10] ms,[96 ± 11] ms vs[ 388 ± 12 ] ms,[379 ± 14]ms),QT dispersion and Tp-Te ( [ 80.4 ± 7.6 ] ms vs [ 78.5 ± 8.3 ] ms) were compared respectively between ERV group and control group and there was no significant difference (P ＞ 0.05).The occurrence of J wave in inferior leads was 67.9％,15.1％for the side wall leads,and 17.0％ for the chest leads,thus inferior wall leads had higher incidence than other hads relativelv in ERV ( P ＜ 0.01 ).Conclusion ERV exhibits more occurrence in inferior wall leads.

ObjectiveTo explore the relationship of postoperative systemic inflammatory response syndrome(SIRS) and preoperative midway through the urine and perioperative renal pelvis urine of percutaneous nephrostolithotomy(PCNL).Methods Participants included 450 patients with urinary calculus who underwent PCNL,preoperative midway through the urine and perioperative renal pelvis urine of PCNL was collected.ResultsOf 450 cases,preoperative midway through the urine germiculture positive 100 cases (22.2％,100/450 ),perioperative renal pelvis urine germiculture positive 85 cases (18.9％,85/450),46 cases ( 10.2％,46/450) occurred SIRS after PCNL.Decompression of perioperative renal pelvis urine germiculture positive 20 cases(23.5％,20/85) preoperative midway through the urine germiculture positive,perioperative renal pelvis urine the bacteria cultures negative 80 cases (21.9％,80/365 ) preoperative midway through the urine germiculture positive(P ＞0.05),preoperative midway through the urine germiculture positive 15 cases ( 15.0％,15/100) in SIRS,preoperative midway through the urine the bacteria cultures negative 31 cases ( 8.9％,31 /350) in SIRS (P ＞ 0.05 ).Decompression of perioperative renal pelvis urine germiculture positive 18 cases (21.2％,18/85) in SIRS,perioperative renal pelvis urine the bacteria cultures negative 28 cases (7.7％,28/365) in SIRS (P ＜ 0.05 ).ConclusionPreoperative midway through the urine has no correlation with the occurrence of SIRS,perioperative renal pelvis urine germiculture positive can predict the occurrence of SIRS,giving corresponding antibiotic treatment can improve the security of PCNL.

The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Gene Expression ; Myocytes, Cardiac ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Transfection

The aim of this study was to observe whether necroptosis is involved in the process of cardiac hypertrophy induced by pressure overload. SD rats underwent transverse abdominal aortic constriction (TAC) operation for establishing cardiac hypertrophy model. The structure and function of the left ventricle of rats were evaluated via echocardiography, left ventricular mass index, the expression of markers of cardiac hypertrophy and histological detection. Real-time PCR and Western blot were used to measure the gene and protein expression of receptor interacting protein kinase 1 and 3 (RIPK1 and RIPK3, the necroptosis markers) respectively. Four weeks after TAC operation, rat model for cardiac hypertrophy was established. The experimental data showed that the gene and protein expressions of RIPK1 and RIPK3 in the rat heart hypertrophic tissues after TAC for 4 weeks were increased significantly compared with those in the sham group. HE staining showed cardiomyocytes injury and hypertrophy in the hearts of TAC rat models. By transmission electron microscope, we observed that mitochondria of cardiomyocytes were damaged seriously in the TAC models. Treatment with losartan used, the selective antagonist of angiotensin II type I receptor could improve the cardiac function of TAC rats. Moreover, losartan treatment decreased the expression of RIPK1 and RIPK3 in heart tissues of TAC rats. The results suggest that necroptosis occurrs in the process of cardiac hypertrophy with pressure overload, and losartan could alleviate the cardiac hypertrophy and inhibit necroptosis.
Animals ; Apoptosis ; Cardiomegaly ; pathology ; Disease Models, Animal ; Echocardiography ; Heart ; physiopathology ; Losartan ; pharmacology ; Myocytes, Cardiac ; Pressure ; Protein-Serine-Threonine Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; metabolism

OBJECTIVE:To explore the effect of Compound pueraria tablets on the kidney tissues of diabetic nephropathy rats. METHODS:High sugar and lipid diet combined with intraperitoneal injection of streptozotocin were used to establish the model of diabetic nephropathy (DN). Normal control group was established. Model rats were randomly divided into model control group, positive control group(Irbesartan tablet),Compound pueraria tablets low-dose,medium-dose and high-dose [0.102,0.203,0.406 g/(kg·d)] groups(n were 8-10). The corresponding drugs were given,and fasting blood glucose(FBG)and 24 h urinary protein (Upro) were collected at 1st,14th,28th,42nd,56th day after treatment. SCr,BUN,TC,TG and KI were detected,and renal pathology was observed after the last dose. RESULTS:Compared with normal group,FBG of those groups were all increased after modeling,and 24 h Upro of them were all increased after 28th day (P<0.05 or P<0.01). Compared with model control group, FBG of Compound pueraria tablets medium-dose and high-dose groups were all decreased since 42nd day (P<0.05 or P<0.01), and 24 h Upro of Compound pueraria tablets groups were all decreased since 28th day(P<0.05 or P<0.01);BUN,TC,TG and KI of Compound pueraria tablets in medium-dose and high-dose groups were all decreased significantly,and SCr of 3 dose groups were all increased (P<0.05 or P<0.01). The morphological structure of renal cells was improved significantly in drug treatment groups. CONCLUSIONS:Compound pueraria tablets can correct lipid metabolism disorder,reduce Upro and improve renal func-tion,indicating certain protective effect on the kidney tissues of diabetic nephropathy rats.

OBJECTIVE:To study the effect of Compound pueraria tablets on osteoporosis model rats induced by retinoic acid. METHODS:The osteoporosis model was induced by intragastric administration of retinoic acid solution for 15 days;normal group was established. After modeling,the rats were randomly divided into model control group,Xianling gubao capsule [0.32 g/(kg·d)] positive control group,Compound pueraria tablets low-dose and high-dose [0.24,0.4 g/(kg·d)] groups (n=8). After 6 weeks of ig,the serum sample was collected to determine the levels of serum calcium(s-Ca),serum phosphorus(s-P),ALP and bone gla protein (BGP);bone density instrument was used to detect the contents of bone mineral density (BMD),bone mineral content (BMC),bone image area (BIA) and muscle content (MC);the results of compact bone substance scanning were observed. RE-SULTS:Compared with normal group,the levels of s-Ca,BMD,BMC and MC in rats were decreased in model control group, while the level of BGP was increased(P<0.05 or P<0.01). Compared with model control group,related index and compact bone substance scanning of Compound pueraria tablets groups were all improved;the levels of s-Ca,s-P,ALP,BMD and MC were in-creased in Compound pueraria tablets high-dose group,while the level of BGP was decreased;the levels of BMD and MC were in-creased significantly in Compound pueraria tablets low-dose group(P<0.05 or P<0.01). CONCLUSIONS:Compound pueraria tab-lets can improve the osteoporosis induced by retinoic acid in rats.