Objective To investigate the relationship between the expression level of platelet membrane glycoprotein?3(GPⅢa,CD61)and the severity of disease in patients with hemorrhagic fever with renal syndrome(HFRS).Methods One hundred and four patients with HFRS and 30 healthy individuals were recruited.The percentage of CD61 positive platelets and the mean fluores- cence intensities(MFI)of platelet membrane glycoprotein?3 were determined by flow cytometry (FCM).The 104 patients studied were divided into three groups based on their expression levels of platelet membrane glycoprotein?3 at oligurie phase.Clinical data and laboratory parameters in different groups were compared and analyzed.Results The expression levels of CD61 in patients with HFRS were significantly higher than those in control group,although no significant difference in the percentage of CD61 positive platelets between patients with HFRS and controls was detected.The MFI of CD61 expression in patients with HFRS at fever phase,oliguric phase and polyuric phase was 19.75?2.57,17.46?1.48 and 15.55?0.60,respectively,which was significantly higher than that in control group(3 20?0.12).The expression level of CD61 in patients with HFRS at oliguric phase was negatively correlated with platelet count and serum albumin(r=-0.637 and-0.695,respec- tively)and positively correlated with white blood cell count,blood urea nitrogen,serum creatinine and alanine aminotransferase(r=0.945,0.904,0.956 and 0.891,respectively).When the patients were compared according to the expression levels of CD61,it was indicated that the higher the expression level of CD61,the higher the incidence of uremia,hypoalbuminemia,abnormal liver func- tion and leukocytosis.Conclusions The expression levels of platelet membrane glycoprotein?3 in patients with HFRS are different in different clinical phases and are significantly correlated with the severity of the disease in the patients.It suggests that the expression levels of platelet?3 integrin are dramatically increased in patients with HFRS,which may be an indicator for the severity of disease and be helpful for monitoring the state of the patients' diseases and evaluating the severity of the disease.

The present study aimed to explore whether the stretch of ischemic myocardium could modulate the electrophysiological characteristics via mechanoelectric feedback (MEF), as well as the effect of phalloidin on the electrophysiological changes. Thirty-two Wistar rats were randomly divided into 4 groups: control group (n=9), phalloidin group (n=7), myocardial infarction (MI) group (n=9), MI + phalloidin group (n=7). The acute myocardial infarction (AMI) was conducted by ligation of the left anterior descending (LAD) coronary artery for 30 min in isolated rat heart. The volume alternation of a water-filled latex balloon in the left ventricle produced the stretch of myocardium. After perfused on Langendorff, the isolated hearts were stretched for 5 s by an inflation of 0.1, 0.2 and 0.3 mL separately and the effect of stretch was observed for 30 s, including the left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), ±dp/dt(max), monophasic action potential duration at 90% repolarization (MAPD90), and occurrence of premature ventricular beats (PVB) and ventricular tachycardia (VT). The stretch caused an increase of MAPD(90) in both control and MI rats (P<0.05, P<0.01). Moreover, MAPD(90) in MI group increased more significantly than that in the control group at the same degree of stretch (P<0.05, P<0.01). Phalloidin (1 μmol/L) had no effect on MAPD(90) in basal state. After stretch, MAPD(90) in phalloidin group slightly increased but was not significantly different from that in the control group. However, phalloidin reduced MAPD(90) in infarcted myocardium, especially when ΔV=0.3 mL (P<0.05). The incidence rates of PVB and VT in MI group were higher than that in the control group (both P<0.01). And there was no significant difference in the incidence rates of PVB and VT between phalloidin group and control group. Phalloidin inhibited the occurrence of PVB and VT in infarcted hearts (both P<0.01). LVSP and +dp/dt(max) in MI group obviously decreased (P<0.01 vs control). With application of phalloidin, LVSP slightly, but not significantly increased in infarcted hearts, while -dp/dt(max) significantly increased (P<0.05). It is suggested that MI facilitates the generation and maintenance of malignant arrhythmias, while phalloidin obviously inhibits the occurrence of arrhythmias.
Action Potentials ; Animals ; Arrhythmias, Cardiac ; prevention & control ; Coronary Vessels ; Heart ; drug effects ; physiopathology ; Heart Ventricles ; Myocardial Infarction ; physiopathology ; Phalloidine ; pharmacology ; Rats ; Rats, Wistar

Adult ; Female ; Hepatitis C, Chronic ; blood ; physiopathology ; Humans ; Leptin ; blood ; Lipoproteins ; blood ; Liver Function Tests ; Male ; Middle Aged

OBJECTIVETo study the validity of single plane Simpson's method with conventional X-ray ventriculography for estimation of right ventricular (RV) volume.

METHODSFifteen human RV casts were obtained from 15 subjects who died from non-cardiac causes within 24 hours after death. These casts were photographed respectively and their volumes were calculated by using the single plane Simpson's method based on a new half-circle model. The actual RV cast volumes were determined by water displacement method.

RESULTSThe actual RV volume was (64.23 +/- 24.51) ml and the calculated volume was (58.04 +/- 24.45) ml. The calculated RV volume underestimated the actual volume by (6.19 +/- 12.38) ml, but there was no significant difference between the actual and the calculated RV volume (P > 0.05). There was a significant correlation between the actual cast volume and the calculated volume (r = 0.983, P < 0.01). The regression equation was: RV actual volume = 1.074 x (RV calculated volume).

CONCLUSIONRV volume calculated by single plane Simpson's method with conventional X-ray ventriculography is accurate and deserves further study.

Adolescent ; Adult ; Aged ; Angiocardiography ; methods ; Cardiac Volume ; Child ; Child, Preschool ; Feasibility Studies ; Female ; Heart Ventricles ; Humans ; Male ; Middle Aged ; Models, Cardiovascular ; Ventricular Function, Right ; X-Rays ; Young Adult

OBJECTIVETo investigate the possible relationship between polymorphism of codon25 in signal peptide region of transforming growth factor beta 1 (TGFb1) and hepatitis C virus (HCV) infection susceptibility.

METHODSGenotypes of TGFb1 of 191 subjects (85 HCV infected patients and 106 healthy controls) were studied. Genotypes of TGFb1 codon25 were determined by amplification refractory mutation system (ARMS).

RESULTSDifferences of codon25 polymorphism were not found between HCV infected patients and the controls (P more than 0.05), which showed a similar pattern between the chronic hepatitis C group and HCV-associated liver cirrhosis group (P more than 0.05). There were no differences of genotype distribution of codon25 between ALT normal and ALT elevated patients (P more than 0.05), but G allele frequency was higher in ALT elevated group (P=0.040). There were great differences between the distribution of genotypes (P=0.005) and allele frequency (P=0.000) of the HCV RNA positive and the negative groups in that the HCV RNA positive group differed greatly from the negative group.

CONCLUSIONPolymorphism of TGFb1 codon25 may influence the grade of liver inflammatory activity. High G allele frequency of codon25 may be associated with viremia in patients with chronic HCV infection. It seems that polymorphism of codon25 in the signal peptide region of TGFb1 may contribute to the outcome of HCV infected patients.

Adult ; Blood Donors ; Case-Control Studies ; Female ; Genotype ; Hepatitis C, Chronic ; genetics ; metabolism ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo discuss the effect of protein kinase C (PKC) on regulation of ecto-5'-nucleotidase activity by lysophosphatidylcholine(LPC) in human umbilical endothelial cells (HUVEC).

METHODSExperiments were conducted in HUVEC grown on dishes which were divided into 4 groups (n=15): (1) Control group in which only eAMP (5 micromol/L) was added; (2) LPC group in which HUVEC were incubated with LPC (10 micromol/L) before eAMP was added; (3) Chelerythrine group in which cells were pre-incubated with the PKC inhibitor chelerythrine (100 micromol/L) before LPC and eAMP were added; (4) alpha, beta-Methyladenosine-5'-Diphosphate (AOPCP) group in which cells were incubated with AOPCP (10 micromol/L) before eAMP was added. Etheno-adenosine production was detected at 15th, 30th, 45th min with high performance liquid chromatography(HPLC) respectively.

RESULTSComparing to the control group LPC significantly increased etheno-adenosine production at three time points respectively (P < 0.05). Furthermore, PKC inhibitor chelerythrine abolished this effect of LPC and the ethenoadenosine production at three time points were at the same level of control group (P > 0.05). CD73 inhibitor AOPCP significantly decreased the etheno-adenosine production compared to the other three groups (P < 0.01).

CONCLUSIONEcto-5'-nucleotidase can be modulated within minutes following exposure of HUVEC to LPC and this response may be mediated by PKC in HUVEC.

5'-Nucleotidase ; metabolism ; Cells, Cultured ; GPI-Linked Proteins ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; Humans ; Lysophosphatidylcholines ; pharmacology ; Protein Kinase C ; physiology ; Up-Regulation ; drug effects

OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.

OBJECTIVE To investigate the protective effect and mechanisms of luteolin-7-O-β-d-glucuronide (LGU) on oxygen glucose deprivation (OGD)-induced H9C2 cardiomyocytes injury. METH-ODS The protective effect of LGU on OGD-induced H9C2 cardiomyocytes death were investigated by MTT assay. The microfilament change of H9C2 cardiomyocytes was detected by phalloidin staining and the lactate dehydrogenase (LDH) leakage rate was also detected by LDH kit. In order to explore the possible mechanisms of LGU, ATP content, intracellular Ca2+fluorescent intensity and concentra-tion, mitochondrial membrane potential (MMP)and the expressions of apoptosis-related proteins were detected by ATP kit,CLSM(Fluo-3/AM probe),Ca2+kit,CLSM(JC-1 probe)and western blotting meth-od, respectively. RESULTS The inhibition of H9C2 cardiomyocyte survival rate inducedby OGD was improvedby pretreated with LGU in a concentrationdependent manner. The microfilaments injury as well as the increase of LDH leakage rate were also improvedby pretreated with LGU.The ATP content was significantly decreased,intracellular Ca2+fluorescent intensity and concentration were significantly increased and the MMP was significantly decreased 4 hafter OGD. LGU significantly reversed the de-crease of intracellular ATP content,the increase of Ca2+fluorescent intensity and concentration and the decrease of MMP.The release of cytochrome C,the expressionsof caspase-9 and caspase-3 in H9C2 cardiomyocytes were increased 16 h after OGD.LGUsignificantly inhibited the changes of these apop-tosis-related proteins. CONCLUSION LGU has a significant protective effect against OGD-induced H9C2 cardiomyocytes injury through inhibiting calcium overload,increasing ATP content,improving mi-tochondrial function and inhibiting apoptosis.

Objective To determine the prevalence and genotype of hepatitis E virus (HEV)among commercial swine population in Eastern and Southern China. Methods Six hundred specimens of swine bile collected from 5 slaughterhouses in Eastern and Southern China from 2007 to 2009 were tested for HEV RNA using nested RT-PCR. PCR products were sequenced for phylogenetic analysis. Results Forty-seven out of the 600 samples (7.83%) were positive for HEV RNA. Based on the 150 nt fragment within HEV ORF2, data from phylogenetic analysis revealed that all the 47 HEV isolates were identified to be genotype Ⅳ, sharing 75.0%-83.4%, 75.0%-84.6%, 71.9%-80.7% and 88.1%-91.5% nucleotide identities with prototype Ⅰ,Ⅱ,Ⅲ and Ⅳ HEV strains respectively while majority of the isolates clustered within their respective isolation sites. Conclusion HEV was widespread in commercial swine population in Eastern and Southern China that raised a serious concern about the safety regarding the consumption of pork products.