Objective To construct a fusion minigene expression vector of seven HCV NS3 epitopes and express the fusion gene in the eukaryotic cell line, in order to form a foundation for the investigation of the DNA immunization for HCV prevention. Methods Three pairs of complementary oligonucleotides primers covering all seven HCV NS3 epitopes were synthesized. Overlapping extension PCR were performed to construct the fusion minigene and cloned in pEGFP-N3 and pBuDCE vector respectively. The fusion protein was detected by Western blotting and flow cytometric analysis. The transfection efficiency was assessed with by flow cytometric analysis. Results The HCV NS3 epitope fusion minigene was obtained and inserted into pEGFP-N3 and pBuDCE vector respectively. The recombinant plasmid pEGFP-DR4 and pBuDCE-DR4 were expressed in 293T and 046W cell lines respectively. The Western blot result indicated that the expected 36 kDa minigene/EGFP fusion product was expressed from pEGFP-DR4, while 13kDa product from pBuDCE-DR4 respectively. Conclusion The HCV NS3 epitope fusion minigene was expressed in the eukaryocyte expression system.

OBJECTIVE To investigate the flora distribution,enzyme producing and drug resistance of Citrobacter in our hospital,and analyzed multi drug resistance(MDR) character in order to guide the clinical medication.METHODS Totally 147 clinical isolates of Citrobacter were detected out ESBLs and AmpC ?-lactamases by three-dimensional test,MBL by the double-disk synergy test.at the same time,drug resistance to fifteen antibiotics was also detected by K-B method.RESULTS Hospital infection caused by Citrobacter.most commonly by C.freundii and then C.amalonaticus.Respiratory tract and urinary tract were prone to be infected than other sites(P0.05).The rate of ESBLs,AmpC,ESBLs+AmpC and MBL which produced by Citrobacter were 36.05% 10.20%,7.48% and 2.72%.The Citrobacter were sensitive to imipenem and meropenem,and their resistance rates to imipenem and meropenem were 4.76% and 3.40%,respectively.The resistance rate to cefoperazone/tazobactam was 23.81%.Otherwise,the resistance rate to 12 kinds of other antibiotics were all higher than 40.0%.MDR strains in the ICU ward were 81.08%,while in other wards were 53.64%,with significant difference(P

Obiective To study the clinic characteristics of young women with breast cancer, and analyse the reason of delayed diagnosis and treatment. Methods A retrospective review was made in 121 cases of young women with breast cancer (age ≤35 years as young group) and 40 cases of breast cancer patients of control group (age≥40 years), who all underwent definitive surgical treatment. Results There were significant differences in the positive rate of invasive tumor and the positive rate of axillary lymph node between the two groups(P

Pulmonary function changes during and after cardiopulmonary bypass were studied using single breath test for CO_2 (SBT-CO_2) in 16 adult patients undergoing cardiac surgery. The results showed that P_(A-a)CO_2 and P_(A-a)O_2 increased significantly after bypass though PaCO_2 kept in normal range by adjusting ventilation volume. The CO_2 production increased as time passed after bypass, resulting in the increase of required minute volume and the rise of airway pressure. Compliance showed a tendency to decrease, while physologic dead space and alveolar dead space increased significantly. It is concluded that the causes of pulmonary dysfunction occurring in early stage of post-bypass are mainly due to the V/Q mismatch induced by low perfusion of the lung. To improve the pulmonary function at the early stage of post-bypass, the circulatory function should be improved accordingly.

Objective To determine the effect of propofol on calcium homeostasis mitochondria in rabbit cardiac muscle subjected to ischemia reperfusion under the protection of cardioplegic solution Methods Twenty four healthy New Zealand white rabbits weighing 1 9 2 2kg were randomly divided into four equal groups of six rabbits each: control group(C);ischemia group(I); low concentration propofol +ischemia(LP) and high concentration propofol +ischemia(HP) The animals were anesthetized with intraperitoneal thiopental 25mg/kg and heparinized Chest was quickly opened and heart excised and connected to Langgendorff preparation The isolated heart was passively perfused via aorta In group LP and HP the perfusate contained propofol 50?mol/L and 200?mol/L respectively When the developed pressure of left ventricle and heat rate were stabilized (about 20 min after perfusion was stared),in group C perfusion was continued for another 90 min at normothermia (37℃?0 2℃)and 7 84kPa perfusion pressure In group I, LP and HP perfusion was stopped and cardiac arrest was induced by 20ml high potassium cardioplegic solution (4℃)and heart was immersed in 8℃ 14℃ normal saline Cardioplegic solution was perfused every 20min After 60 min cardiac arrest the heart was reperfused for another 30min The heart was then removed and immediately kept in 0℃ hemogeneous medium A slice of cardiac muscle was taken from apex of the heart Mitochondrial structure was observed using electron microscope Mitochondrial matrix calcium concentration, mitochondrial calcium uptake in high calcium enviroment (2 5 mmol/L CaCl 2 was added ) and calcium release (when sodium ion was added) were determined Results Propofol at concentration of either 50?mol/L or 200?mol/L had no effect on mitochondrial calcium homeostasis after ischemia reperfusion The damage to mitochondrial structure in propofol groups was milder than that in ischemia group Conclusions Propofol at clinical concentration can relieve the mitochondrial permeability transition pore opening at reperfusion stage The mechanism is not through inhibition of mitochondrial calcium overload, probably a result of diminished oxidative stress

Objective To establish a rat model of bone cancer pain by intra-tibia inoculation of Walker 256 mammary gland carcinoma cells. Methods Sixty female Wistar rots weighing 180-200 g were randomly divided into 4 groups (a=15each):groupⅠ normal control; group Ⅱsham operation; group Ⅲtumor cell inoculation + normal saline (NS) and group Ⅳtumor cell inoculation + flurbiprofen. NS 0.2 nd and flurbiprofen 10 mg/kg in 0.2 ml were injected IV at 2 h before determination of pain threshold on 14 and 17 d after inoculation oftumor cells in groupⅢand Ⅳ respectively. On day 0, 4, 7, 10, 14, 17 and 21 after inoculation pain threshold was measured after determination of body weight. X-ray examination of the tibia was performed on day 14 after inoculation. The animals were killed on day 21 after inoculation for microscopic examination of the inoculated tibia. Results The animals started losing weight and the threshold to yon Frey hair stimulation was decreased from dhy 10 after inoculation in group Ⅲand Ⅳ. X-ray examination showed destruction of bone and microscopic examination showed tumor growing in tibia. Flurbiprofen significantly decreased mechanical hyperalgesia in group Ⅳ. There was no significant difference in paw withdrawal latoney to radiant heat among the 4 groups. Conclusion A model of bone cancer pain can be made by inoculation of Walker 256 mammary gland carcinoma cells into tibia characterized by mechanical hyperalgesia.

Objective To observe the effect of fluvastatin on urinary albumin excretion rate(UAER)of microalbuminuria in type 2 diabetes patients.Methods Type 2 diabetes patients with mieroalbuminuria were randomly divided into fluvastatin goup(n=126)and control group(n=129).Fluvastatin group was given fluvastatin 40 mg each night.Follow up was conducted for 1.5 years.The UAER and glomerular filtration rate(GFR)as well as blood lipid level before and after therapy were compared.Results UAER in fluvastatin group was significantly lower than that in control group[(59.6±10.5)vs(87.5±12.3)mg/min,P＜0.05]before therapy[(104.4±25.2)vs(110.6±19.7)mg/min,P＜0.05],which was independent of its lowering-lipid effect.But GFR had no signifimicroalbuminuria which is independent of lowering-lipid effect in type 2 diabetic nephropathy patients and delay the progress of diabetic nephropathy.

Objective To define the factors that influence the safety of hepatectomy for hepatocellular carcinoma (HCC) associated with cirrhosis. Methods Based on the patients treated before and after February 1997,229 cases of HCC associated with cirrhosis were divided into two groups, Group A and Group B, respectively. The patients′ general condition, operative procedure, morbidity and mortality rates were compared between the two groups. The factors that influenced surgical morbidity were analysed. Results In group B, patients′ average age was higher (P

Objective To evaluate the nutritional status of extremely low birth weight (ELBW) infants and the effects of nutritional support strategy alterations on their growth during hospitalization.Method From 2005 to 2014,clinical data of ELBW infants admitted to the neonatal intensive care unit (NICU) in our hospital were retrospectively analyzed.The clinical data included their general status,enteral and parental nutritional support strategy and complications during hospitalization The patients were assigned into pre5 group and late5 group.Those who survived and discharged from 2005 to 2009 were the pre5 group,and those who survived and discharged fromn 2010 to 2014 were the late5 group.The independent t test and chi square test were used for statistical analysis.Result A total of 58 ELBW infants were enrolled in the study,including 18 patients in the pre5 group and 40 in the late5 group.No statistically significant differences existed between the two groups on gestational age,birth weight,Z score (weight for length and gender),birth length,head circumference and main complications during hospitalization (P ＞0.05).Pre5 group had higher incidence of small for gestational age (SGA) than late5 group (16/18 vs.25/40,P =0.037),while the EUGR ratio at discharge (14/18 vs.21/40,P =0.061) was similar.When compared with pre5 group,late5 group had larger amount of initial enteral feeding volume [4.4 ml/(kg · d) vs.2.4 ml/(kg · d),P =0.014] and feeding volume at the end of the first week [(19.8 ± 16.0) ml/(kg · d) vs.(12.2 ±9.5) ml/(kg · d),P =0.036].Similarly,the starting dose of amino acids in parenteral nutrition [2.0g/(kg· d) vs.1.0 g/(kg· d),P＜0.001],maximum dose of amino acids [4.0g/(kg.d) vs.3.5 g/(kg · d),P ＜ 0.001],total calories at the end of the first week [(82.6 ± 12.6) kcal/(kg · d) vs.(71.1±15.2) kcal/(kg· d),P=0.004] and the second week [(103.7 ±19.8) kcal/(kg· d) vs.(92.3 ± 17.9) kcal/(kg · d),P =0.041],the weight gain velocity from birth to discharge [(18.7 ± 2.9) g/(kg.d) vs.(16.9±2.8) g/(kg· d),P=0.031] and change of Z scores (AZ) [-0.6 (-1.0,-0.4) vs.-1.2 (-1.6,-0.8),P =0.004] showed significantl differences between the two groups,with better outcomes in late5 group.However,the duration of parenteral nutrition,the total amount of amino acids,the time reaching total enteral feeding,the length of hospital stay were similar between the two groups.Ten cases(61.1％)of infants in pre5 group were breastfed,and four of them used human milk fortifier (HMF) (fortified rate was 22％).32 cases (80％) in late5 group were breastfed and 23 cases used HMF (fortified rate was 57.5％).The time to initiate HMF in the late5 group was at (30.2 ± 13.2) days,and human milk amount was (89.9 ± 34.5) ml/kg,fortified duration was (32.8 ± 15.7) days.Conclusion The enteral feeding strategy were more vigorous in the last 5 years than before,the initial feeding volume,the increasing rate,the initial dosage of amino acid,and maximum dosage of amino acid had been increased.Human milk and HMF of preterm infants were preferred.The vigorous nutritional support strategy were effective for the weight gain of ELBW infants without obvious side effects during hospitalization.

Objective:To study heart mitochondrial calcium homeostasis during ischemia- reperfusion under the protection of cardioplegic solution.Methods:Isolated perfused rabbits hearts were subjected to global ischem ia in the presence of cold cardioplegic solution for an hour and reperfused for half an hour.Electron m icroscope was used to observe the change of mitochondrial structure.Heart m itochondria was isolated and Fura2 - AM was used as the indicator of matrix concentration of calcium([Ca2 + ]m) to m easure[Ca2 + ]m,the intake and outtake ability of the calcium when m itochondria was at high calcium concentration liquid or high natrium concentration liquid.Results:[Ca2 + ]m increased significantly but mitochondria was still able to uptake calcium and release calcium of m atrix.There was some electron- dense deposit in mitochondria and the injury of mitochondria could be seen by electron microscope.Conclusion:The reason for mitochondria calcium overload at the beginning of reperfusion include physiological and pathological calcium uptake,which ends after30 min reperfusion under the protection of cardioplegic solution. [