Objective To study the persistence and replication of hepatitis C virus (HCV) RNA in Epstein Barr virus (EBV) transforming peripheral blood mononuclear cell (PBMC) from patients with he patitis C as well as variation of hypervariable regions (HVR) of the HCV genome. Methods PBMC from one patient with hepatitis C was infected by EBV and then transformed into lymphoblasts capable of being propagated indefinitely. Then, HCV RNA of the cultured cells and supernatants was detected by reverse transcriptase polymerase chain reaction (RT PCR) every month and in situ PCR to identify the location of HCV RNA in the cells. The HVR genome sequences of HCV in the first and the ninth month subculture cells were identified by sequence analysis. Results HCV plus strand RNA could be detected in the cultured cells for as long as 1 year. The HCV plus strand RNA could be identified in supernatants and the minus strand RNA were also observed in the cultured cells intermittently. In situ PCR showed that the blue black positive signals of HCV RNA located mainly in cytoplasmas of EBV transforming B cell but negative in nucleous. The positive signal was not found in negative control cells by in situ RT PCR. HCV HVR genome sequence after half year was found to have two regions of a high degree of variability in 1491 to 1583 nucleotide (384 to 414 amino acid) and 1761～1781 nucleotide (473 to 479 amino acid). But, compared the HCV HVR genome sequence in first month subcultured cells with that in ninth month, there was not significant difference. Conclusion HCV may exist in the cultured cell line for a prolonged period wihtout HVR genome sequence changed.

Objective To investigate the effect of various concentration of inhaled oxygen during exercise on reactive oxygen species (ROS) production in chronic obstructive pulmonary disease (COPD) rats and the possible mechanism. Methods Eighty COPD Wistar rats were divided into low oxygen (LO, n=20), normal oxygen (NO, n=20) and inhaled oxygen (IO, n=20) exercising groups, which ran on tread-mill in the conditions of 13.6%, 21%, 25%oxygen, respectively, and non-exercising normal oxygen group (C, n=20), which stood on still treadmill in the condition of 21%oxygen. Their apoptosis percentage of neutrophils and ROS content were measured with flow cytometry, glutathione (GSH) in lung with immunohistochemistry and cytochrome C oxidase IV (COXIV) in skeletal muscle with Western blotting, as one and four weeks of exercising. Results For one week of exercise, the ROS was more in LO and NO groups than in C group (P<0.05), while the neutrophils apoptosis percentage was less (P<0.05);and there was no difference among C, LO, NO, IO groups in expression of GSH and COXIV (P>0.05). For four weeks of exercise, the ROS was more in LO group but less in NO and IO groups than in C group (P<0.05);while the expression of GSH and COXIV increased in NO and IO groups compared with those in C group (P<0.05). Conclusion A long term exercise in non-hypoxic state may inhibit ROS production for COPD patients, by promoting neutrophils apoptosis and antioxidant expression.

Objective To determine the effects of intrauterine growth retardation(IUGR) caused by malnutrition during pregnancy on the lung structure and expression of Clara cell protein (CCSP) and interferon (IFN)-γ in the fetal lungs,and to explore their relation ship with pulmonary disease.Methods Fetal rats from maternal protein-malnutrition dams were studied on day 20(term 21.5 day).The lung pathology was examined by means of Hematoxylin and eosin(HE) stain.Plasma was collected to determine the CCSP and IFN-γ concentration.Lungs were harvested to measure the expression of CCSP and IFN-γ mRNA by using fluorescent quantization reverse transcription (RT)-PCR and the levels of CCSP and IFN-γ protein were assessed by using enzyme-linked immunosorbent assay.Results Malnutrition fetus body weight significantly less compared to control group,so did the lung weight.However,lung weight,expressed as a percentage of body weight between the 2 groups was not different.The IUGR group had significantly decreased alveolar number manifested by lower radial alveolar count,and significantly increased mean linear intercept of alveoli.Both the CCSP mRNA expression and protein level in lung of IUGR rats were decreased compared with control rats (all P ＜ 0.05).A decline in plasma CCSP protein concentration was also noted compared with control group (P ＜0.05).Furthermore,IUGR group fetus showed lower IFN-γ levels both in circulation and in the lung tissue (all P ＜ 0.05).Conclusions Intrauterine malnutrition significantly alters lung structure and cytokine IFN-γ level,and the latter may further inhibit the transcription of CCSP gene.These alterations may contribute to both early and late postnatal respiratory morbidity.

Objective To evaluate the role of P2X receptors in the synthesis and release of IL-1β during oxygen-glucose deprivation (OGD) in rat hippocampus. Methods Male SD rats weighing 150-200 g were anesthetized with ether and decapitated. The hippocampi were removed and sagittally sliced (400 μm thick) and placed in artificial cerebrospinal fluid (aCSF) aerated with 95% O2-5% CO2. One hundred and sixty hippocampal slices were randomly divided into 4 groups ( n = 40 each): Ⅰ control group (group C); Ⅱ OGD group; Ⅲ OGD +BBG group; Ⅳ OGD + anti-P2X4 group (group OP). In group C, the hippocampal slices were continously incubated with aCSF aerated with 95% O2-5% CO2 . In group OGD, the hippocampal slices were incubated with glucose-free aCSF and aerated with 95% N2-5%CO2 . In group OGD + BBG, the hippocampal slices were incubated with aCSF containning P2X7 receptor-specific antagonist G (BBG, final concentration 1 μmol/L) and aerated with 95% O2-5% CO2 for 20 min, then exposed to OGD, and BBG (final concentration 1 μ mol/L) was added in glucose-free aCSF. In group OP, the hippocampal slices were incubated with aCSF containning P2X4 receptor antibody (final concentration 1.5 μg/ml) and aerated with 95% O2-5% CO2 for 60 min, then exposed to OGD, and P2X4 receptor antibody (final concentration 1.5 μg/ml) was added in glucose-free aCSF. LDH and IL-1β release was detected before OGD and at 20, 40 and 60 min of OGD. Histological changes were observed using HE staining.Intracellular pro-IL-1β precursor protein expression was detected by Western blot at 60 min of OGD. Results LDH and IL-1β release and expression of intracellular pro-IL-1β precursor protein were signifcantly higher in the other groups than in group C ( P ＜ 0.05 ). Compared with group OGD, LDH and IL- 1 β release was signifcantly decreased, while the expression of intracellular pro-IL-1β precursor protein un-regulated in group OGD + BBG ( P ＜0.05), but no signifcant difference was found in the prarameters mentioned above in group OP ( P ＞ 0.05). Conclusion P2X7 receptor mediates the synthesis and release of IL-1β during OGD in rat hippocampus, but P2X4 receptor does not.

Objective To evaluate the role of P2X7 receptors in release of glutamate (Glu) and γ-aminobutyric acid (GABA) during oxygen-glucose deprivation (OGD) in rat hippocampus and neuronal synaptosome.Methods Healthy male SD rats weighing 150-200 g were decapitated. Their hippocampi were isolated and cut into slices 400 μm thick or made into neuronal synaptosomes. The hippocampal slices and neuronal synaptosomes were incubated in artificial cerebro-spinal fluid (aCSF) at 35℃ for 30 min and divided into 3 groups ( n = 32 or 24 each): control group (group C); group OGD and group OGD + BBG (brilliant blue G, a specific P2X7 receptor antagonist). OGD was induced by incubating the slices and synaptosomes in glucose-free aCSF aerated with 95% N2-5% CO2. In group OGD + BBG the slices and synaptosomes were incubated in O2-glucose deprived aCSF containing BBG 1 μmol/L 2 ml. Release of Glu and GABA from hippocampal slices and synaptosomes was determined by HPLC at 0, 20, 40, 60 min of OGD (T1-4). Hippocampal slices were examined with microscope.Results ( 1 ) The release of Glu and GABA from hippocampal slices and synaptosomes were significantly increased after OGD ( P ＜ 0.05). (2) Glu released from hippocampal slices was significantly decreased at T3-4 and Glu released from synaptosomes increased at T2-4 in group OGD + BBG as compared with group OGD ( P ＜ 0.05). (3)GABA released from hippocampal slices was significantly decreased at T4 in group OGD + BBG as compared with group OGD ( P ＜ 0.05). There was no significant difference in GABA released from synaptosomes between group OGD and OGD + BBG (P ＞ 0.05). (4) Microscopic examination showed that OGD induced significant histopathological damage to hippocampal slices which was attenuated by BBG treatment. Conclusion P2X7 receptors mediates the release of Glu and GABA during OGD in rat hippocampus and the P2X7 receptors in glial cells plays a leading role.

Cultivating students' creative spirit was an important aim of higher education,and improving scientific research ability was an effective way to realize this aim.Using Journal of Zhejiang Medical College as the carrier,Journal editorial department tried to improve students' scientific research ability.For instance,served fund projects,found competition's added value,offered course about research design and paper writing.The students' scientific research ability was improved.

Objective To explore the electrophysiological features of triggering atrial premature contraction(APC) in patients with hypertension combined paroxysmal atrial fibrillation (PAf).Metbods The originating sites and prematurity index (PI) of triggering APC were analyzed in 46 PAf patients with hypertension (hypertension group)and in 35 PAf patients without hypertension( non-hypertension group)from April,2008 to March,2011 in Shanghai Punan Hospital of Pudong New District.Results Triggering APCs in 46 cases with hypertension combined PAf in the hypertension group originated mainly in the left atrium( 81.6％ ).The coupling interval (CI)of triggering APC in hypertension group was significantly shorter than that in non-triggering APC ( [ 374.1 ± 31.5 ] ms versus [ 443.6 ± 32.6 ] ms,t =23.361,P ＜ 0.001 ) and that in triggering APC in nonhypertensive group ( [374.1 ±31.5]ms versus [395.7 ±38.2]ms,t =5.549,P ＜0.001 ).PI in triggering APC was lower than that in non-triggering APC in hypertension group(0.50 +0.05 versus 0.63 ±0.06,t =22.544,P ＜ 0.001 ) and that in triggering APC in non-hypertension group (0.50 + 0.05 versus 0.55 ± 0.08,t =5.849,P ＜ 0.001 ).Conclusion The triggering APC in patients with hypertension combined PAf mainly originates in the left atrium,the PI of triggering APC is significantly lower than that in non-hypertension patients with PAf and PAf occurs more easily in patients with hypertension.Prompt measures should be taken for hypertension patients with lower prematurity index to prevent the occurrence of PAf.

Objective To learn birth weight of newborn babies and its influencing factors to provide evidence for maternal healthcare.Methods A retrospective analysis was conducted among 5 055 newborn babies from January 2007 to December 2007 to learn their birth weight,gender,pregnant week,maternal age,parity and living areas.Univariate and multivariate Logistic regression analysis were used to explore factors that influenced birth weight.Results For those 5 055 infants,the average birth weight was (3128±675) g,and the incidence of low birth weight or fetal macrosomia was 14.6％ and 8.2％,respectively.In single factor Chi-square test,living area,pregnant week and maternal age were factors that influenced low birth weight and fetal macrosomia (x2 values were 223.807 and 34.120; 2 211.570 and 68.941; 92.199 and 18.745,respectively).Moreover,parity was related with the occurrence of low birth weight (x2=54.822),and gender was found to affect fetal macrosomia (x2=34.503,both P＜0.05).In Logistic regression analysis,preterm birth (odds ratio (OR)=37.140,95％ confidence interval (CI):30.094-45.853),rural residents (OR=0.390,95％ CI:0.310-0.492) and maternal age (OR=0.864,95％ CI:0.779-0.959) were risk factors of low birth weight,and baby boy (OR=0.524,95％ CI:0.423-0.650),urban residents (OR=0.616,95％ CI:0.501-0.758),postterm delivery (OR=4.175,95％ CI:2.918-5.974) and advanced maternal age (OR=1.229,95％ CI:1.104-1.368) were risk factors of fetal macrosomia.Conclusion This investigation suggests a relatively lower average birth weight and higher incidence of fetal macrosomia and low birth weight infant in Changzhi of Shanxi Province.Health interventions,maternal healthcare service programme and pre-and post-natal health education should be carried out to achieve normal birth weight.

125 strains of the symbiotic and epiphyte microorganisms were isolated from marine organisms (sea cucumber, sea urchin,anemone, sea actinia, Ulra, Sargassum, Undaria). Among them,21 strains of bacteria,8 strains of actinomycetes and 2 strains of fungi showed antagonistic activity on bacterial or fungal growth. In the 21 strains of bacteria, 7 strains belong to Bacillus sp., 11 strains Vibrio sp., and 3 strains Pseudomonas sp.. In the 8 strains of actinomycetes, 5 strains belong to Streptomyces sp., 3 strains Micromonospora sp., 2 strains fungi Penicillum sp..

Objective To evaluate the impact of positive end-expiratory pressure (PEEP) on hemodynamic variables including central venous pressure (CVP),mean arterial pressure (MAP) and heart rate (HR) in patients with central respiratory failure treated by mechanical ventilation.Methods Thirty two neurosurgical patients with central respiratory failure,male 19,female 13,mean age (58.8 ± 13.9)years,GCS ≤ 8 points,were enrolled in this prospective,self-control study between June 1,2009,and May 31,2011.Patients were excluded in cases of severe cardiopulmonary disorders,pericardial diseases,person machine confrontation,administration of vasoactive drugs,and hypervolemia or hypovolemia.On admission to neurosurgical intensive care unit,all patients were mechanically ventilated in the mode of synchronized intermittent mandatory ventilation.Hemodynamic effects of six randomly set levels of end-expiratory pressures (0,3,6,9,12,15 cm H2O,every 10 min,1cm H2O =0.098 kPa) were studied in all patients.CVP,MAP and HR were recorded at each of the six end-expiratory pressure levels.One-way analysis of variance and simple linear regression model were used for data analysis.Results The levels of central venous pressure were elevated with increase in end-expiratory pressures.CVP levels were positively correlated with the levels of PEEP (R =0.468,P =0.000),with a simple linear regression equation expressed as:CVP (cm H2O) =7.870 +0.344 ×PEEP (cm H2O),The levels of MAP showed no statistically significant changes at different PEEP levels (F =1.390,P =0.227).No linear correlation between MAP,HR and PEEP levels was found (R =0.042 and 0.160,P =0.413 and 0.002).Conclusions CVP values would be overestimated during mechanical ventilation at different PEEP levels in mechanically ventilated patients due to central respiratory failure,positive correlation existed between CVP values and PEEP levels,whereas MAP was unaffected by different PEEP levels.This study could probably offer a quantitative reference for correct assessment of such a hemodynamic variable as CVP for mechanically ventilated patients without discontinuance of PEEP.Further studies are needed to determine whether these findings could be confirmed in a prospective manner.