Objective To analyze the HEV genetic structure and nucleotide homology isolated from sporadic acute hepatitis E in Changchun.Methods HEV RNA was detected with reverse transcription nested polymerase(RT-nPCR) and sequenced by automatic sequenator.Results The length of acquisitive HEV-CCC220 was 7193 bp coding 2397 amino acid.The gene of HEV-CCC220 was composed of 5'-UTR(nt 1-9),3'UTR(nt 7152-7193) and three open reading frames(ORFs).Its homology with HEV IV was obviously higher than that with HEV(I-III.) The whole genome sequence nucleotide homology was 83.2%-85.0%,85.0%-93.9% and 80.6%-86.7% as compared with 300nt and 98nt partial nucleotide sequence of ORF2.The homology comparison of various areas and various lengths of nucleotide was different.Conclusion The HEV-ccc220,which sporadically happened in Changchun,is the subtype IV of HEV.The 300nt of ORF2 can replace the whole gene order to perform genotype analysis of HEV.

Objective: To verify 5-HT4 receptor mRNA expression in intestinal mucosa mast cells (IMMC). Methods: IMMC was isolated from the whole intestines of normal rats by collagenase digestion and purified by percoll. In situ hybridization was performed to detect the expression of 5-HT4 receptor mRNA in IMMC. Results: Evidently positive staining was shown in the cytoplasm of IMMC. Conclusion: We have verified the expression of 5-HT4 receptor mRNA in IMMC for the first time and provided evidence for further research.

Objective To study the role of gap junctions in epileptiform activity. Methods The epileptiform activity was induced by zero-Mg 2+ medium in cultured hippocampal neurons of newborn rats. Immunocytochemistry and real time RT-PCR were introduced to evaluate the expression of gap junction Cx32 and Cx43. Results The level of Cx32 mRNA increased quickly one hour after the neurons were treated with zero-Mg 2+ medium and was raised by 10 times 5 hours later, while Cx32 protein began to develop at the 2nd hour (21.80?1.74) and was raised by 5 times at the 8th hour (47.30?5.75). The expression of Cx43 mRNA went up obviously 5 hours later, and Cx43 protein developed visibly 8 hours later. Carbenoxolone depressed the expressions of Cx32 and Cx43. Conclusions The expression of Cx32 and Cx43 increases dramatically after epileptic discharges and carbenoxolone inhibits both the discharges and the expression of gap junctions, which indicates that gap junction could contribute to epileptogenesis.

We report a case of symptomatic bradycardia caused by consumption of a Chinese herbal medicine which was initially undisclosed to the attending emergency physician. The scientific name of the herb is Panax japonicus. Electrocardiogram revealed sinus bradycardia. Laboratory tests were normal except for the detection of a high serum digoxin level. Further interrogation of the patient eventually disclosed ingestion of the herb which, however, did not contain any digoxin. Other active ingredients in the herb include various types of ginsenoside. These are digoxin-like substances that had caused the observed false-positive detection of digoxin by fluorescence polarization immunoassay due to cross-reactivity. Our case-report provides an important insight about a blind-spot in the field of laboratory medicine (clinical pathology), namely, the false positive detection of digoxin due to crossreactivity in the immunoassay when we come across digoxin-like substances in clinical scenarios, which has barely received attention in the medical literature. It also conveys a clear educational message that with full understanding of the laboratory methodology and its mechanistic rationale there are actually some tricks-of-the-trade that allow us to optimize the specificity of the biochemical tests and the treatment of digoxin-like substances overdose.

Objective To study the polymorphism of human platelet antigen HPA-1 to HPA-5,and HPA-15 system in Qingdao Han population.Methods A total of 918 samples from regular voluntary platelet donors in Qingdao were genotyped for HPA-1 to-5 and HPA-15 by PCR-SSP.Results The gene frequencies of HPA-1a,-1b;HPA-2a,-2b;HPA-3a,-3b;HPA-4a,-4b;HPA-5a,-5b;HPA-15a,-15b were 0.9940,0.0060;0.9319,0.0681;0.5822,0.4178;0.9897,0.0104;0.9804,0.0196;0.4913,0.5087,respectively.Both a and b alleles were found in each of the 6 HPA systems,and a/a homozygosity was more common in HPA-1,-2,-4 and-5 systems.The HPA genotype frequencies followed Hardy-Weinberg principle.HPA-1 frequency of Qingdao people was significantly different from that of North China(P

OBJECTIVE:To establish a method for contents determination of rutin,quercetin and total flavonoids in Buck-wheat,and compare the difference of flavone in buckwheat in different varieties and areas. METHODS:HPLC was adopted for content determination of rutin and quercetin:the column was Aglient C18 with mobile phase of methanol-0.4%phosphoric acid(gra-dient elution)at a flow rate of 1 ml/min,detection wavelength was 360 nm,column temperature was 40 ℃,and injection volume was 10 μl;UV spectrophotometry was adopted for content determination of total flavonoids:reference solution was methanol,de-tection wavelength was 360 nm,the standard was rutin. Statistical method was used to analyze the differences among the Fagopy-rum tataricum,F. esculentum and flavonoids of F. tataricum from different areas. RESULTS:The linear range was 0.400 9-16.03 μg for rutin (r=0.999 4) and 0.009 9-0.396 0 μg for quercetin (r=0.999 7),RSDs of precision,stability and reproducibility tests were lower than 2%,recoveries were 99.33%-104.00%(RSD=1.32%,n=9) and 96.92%-101.66%(RSD=1.60%,n=9),re-spectively. The linear range of total flavonoids(recovded by rutin)was 4.14-41.4 mg/L(r=0.999 9),RSDs of precision,stability and reproducibility tests were lower than 2,and recovery was 96.07%-101.96%(RSD=2.63%,n=9). The content of flavonoids on F. esculentum is significantly higher than in F. tataricum from the same area,and F. tataricum in Bijie area is better than other places. CONCLUSIONS:The method is simple,stable,reproducible,and can be used for the contents determination of rutin, quercetin and total flavonoids in F. esculentum. The study can provide theoretical basis for developing and using buckwheat.

[Objective] To investigate the construction and inhibitory effects of artificial microRNA targeted on hepatitis B virus.[Methods] Four artificial microRNA sequences for HBV gene were designed with Invitrogen online design software (BLOCK-iTTM RNAi Designer),amiRNA eukaryotic expression plasmid were constructed and transfected into human hepatocellular carcinoma cell line HepG2.2.15.Hepatitis B virus DNA in supernatant was detected by quantitative fluorescence PCR.HBsAg and HBeAg detected by ELISA.[Results] All of the four artificial microRNA sequences could down regulate the level of HBV-DNA,HBsAg and HBeAg.miR3 had the most significant effect on inhibiting HBV-DNA replication.[Conclusion] Artificial microRNA sequences can effectively inhibit the replication of HBV in vitro.

OBJECTIVE: Epileptic attack can cause neuronal damage and increase the risk of potential seizure. Analysis of the possible mechanism of neuronal damage following epileptic seizure may provide evidences for implementing preventive measures against brain damage due to epileptic seizures.DATA SOURCES: A computer-based search of the related publications in PubMed database between June 1995 and June 2004 with different combinations of the key words of "epilepsy", "neuron damage", "necrosis"and "apoptosis", limiting the results to the language of English.STUDY SELECTION: The retrieved articles were examined at first to select reports of experimental study on human and animals related to epilepsy and the subsequent neuronal damages, and their full-text publications were obtained with the other unrelated articles excluded.DATA EXTRACTION: Eighteen articles documenting randomized controlled experiment immediately related to neuronal damage after epilepsy seizure, 4 reporting non-randomized controlled experiments related to central neuronal excitatory toxic damage, and 3 concerning neuronal damage were collected for this review.DATA SYNTHESIS: In the 14 randomized controlled experiments, chemical or electric methods were used to induce epilepsy in the animal models in which the ultrastructural changes of the neurons and cell organelles were observed and the expression of apoptosis-related factors determined.In the 4 non-randomized controlled experiments, central neuronal ischemic and hypoxic models were adopted for observing the expression of various apoptotic factors in the neurons due to different damages with the assistance of electron microscope, to provide direct evidences for the mechanism of central neuronal excitatory toxic damage. The other three related literatures introduced the pathways of neuronal damages and the expression of the related factors.CONCLUSION: Neuronal death after epileptic seizure is correlated with the severity of the damage and mitochondrial functional status, and the mitochondria constitute the control center for neuronal survival. The release of cytochrome C and the activation of caspases are the final common pathway of neuronal damage.

To investigate the neurepretoctive effect of the aqueous extract of dried fruits of lycium barbarum(LBA)OH the hereditary retihal dystrophy of the Royal CoUege of Surgeon(RCS)rats,a total of 24 postnatal RCS mrs were randomly and equally divided into two groups.control group(CG)and experimental group(EG),respectively.EG rats were fed with 1 mg/kg LBA daily 10 days postpartum,and CG rats were fed normally.CG and EG RCS rats were executed at postnatal 25,35 and 50 days.Then photoreceptor cells necrosis and apoptosis Wag assessed by HE staining,and TUNEL detection.The expression of Cagpage-2 protein Wag detected in retina by immunohistochemistry.The results showed that comparing with RCS rata in the CG,25 and 35 days postpartum,the positive expression of TUNEL in RCS rats in the EG obviously reduced(P＜0.05);25 and 50 days postpartum,Caspage-2 positive ceils were seen in both ganglion cell layer and inner nuclear layer in both CG and EG rats.The positive expression of Caspage-2 Wag less$een 25 days postpartum in EG rats than CG rats(P＜0.05).The present results suggest that LBA hag neuroprotecdve effects on the early stage of retinal degeneration of the hereditary retinal dystrophy RCS rats by inhibiting apoptosis,resulting more photoreceptors preserved.Cagpage-2 protein may contribute to the retinal degeneration and LBA might protect apoptotic ueuron$by inhibiting the expression of Cagpage-2.

Objective The purpose of this study was to compare the differences of job satisfaction degree between contract nurses and formal nurses. Methods 350 incumbency clinical nurses were ob-tained from 3 provincial and grade three A hospitals(including nurses in internal medicine, surgical,gyne-cologic,pediatric,emergency departments and ICU). Research instruments used in this study included the self-designed demographic questionnaire and the nurses' job satisfaction scale. Results The level of the overall job satisfaction degree among contract nurses was lower than that of the formal nurses. Of all job satisfaction subscales, the scores of professional opportunities, performance and responsibility,salary and welfare, administration and hospital policy, interpersonal relationships among contract nurses were lower than those of formal nurses. But the scores of scheduling and work conditions, professional traits were higher than those of formal nurses. Conclusions It is advocated to adopt different management and incentive mechanisms to improve the job satisfaction degree for contract nurses according to the differ- ences between contract nurses and formal nurses, so that the nursing staff can be stabilized.