Objectlve To investigate the inhibition of Coxsackievirus B3(CVB3)infection in cardiac myocytes cultured by small interfering RNA(siRNA)-mediated RNA interference and to evaluate the feasibility of siRNA as the prophylaxis and therapy for CVB3 infection.Methods Cardiac myocytes were prepared in vitro and infected with CVB3,and transfected with siRNA by lipofectamin and electroporation.The numbers of beating cardiac myocytes were counted under the microscope.Neutral red staining was used to evaluate the mortality of cardiac myocytes.Antiviral activities of these siRNAs were estimated by observing cytopathic effect(CPE),plaque reduction assay,Western blot assay and RT-PCR.Results siRNA-3753,which aimed at sequence in 2B section of CVB3 genome,displayed a stronger inhibition of CVB3 infection through screening in HeLa cells,siRNA-3753,chosen to transfect cultured neonatal mice cardiac myocytes,Wag observed to keep a good states of growing and beating at 24 h after CVB3 infection.Whereas the cytopathic signature of controlled cells became stopping beating,round and finally the cell fell off the culture plate.The results showed that siRNA-3753 could protect cells significantly,98.1%inhibition of CVB3 replication with electroporation transfection and 78.2%inhibition of CVB3 with liposome transfection.Transfection efficiencies were 56.0 3%and 9.0%by electroporation and lipofectamin,respectively.Conclusion siRNA,which aims at sequence in 2B section of CVB3 genome,can inhibit CVB3 infection in cultured cardiac myocytes.

BACKGROUNDThe heavy incidence and mortality of hemorrhagic fever with renal syndrome, as well as no specific drugs in curing the disease, clearly indicate the need for development of the more effective hantavirus vaccine. Refining the DNA vaccination strategy to elicit more clinically efficacious immune responses is now under intensive investigation. In the present study, we examined the effects of using an interleukin-12 expression plasmid as a genetic adjuvant to enhance the immune responses induced by a DNA vaccine based on the S gene encoding nucleocapsid protein against hantavirus.

METHODSBALB/c mice were immunized three times by intramuscular inoculations of DNA vaccine encoding of hantanvirus nucleocapsid protein alone or in combination with a plasmid expressing murine interleukin-12 (pcIL-12). Booster immunizations were employed 2 times at 2-week interval. To evaluate the humoral and cellular immune responses, antigen-specific lymphocyte proliferation and antibody production were assayed by MTT method and ELISA respectively. The level of interleukin-4 and interferon-gamma in the splenic lymphocytic cultured supernatant were detected with ELISA kit at day 5, 10, 17, 35 and 42 after primary immunization.

RESULTSAntigen-specific IgG antibodies was increased markedly at day 17 in the experiment groups and reached a plateau after day 35. As pcIL-12 co-injected, a significant inhibition of antigen-specific IgG levels was displayed over the period and the antibody mean titre was decreased to only about 1:50 at day 42 after primary immunization, significantly lower than the group immunized with pcDNA3.1 + S alone, in which the mean titre was about 1:70. Interferon-gamma was increased remarkably by the co-injection of pcIL-12 compared with the injection of pcDNA3.1 + S alone. However, the production of interleukin-4 was inhibited by pcIL-12 co-injection. Furthermore, pcIL-12 co-injection efficiently enhanced antigen-specific lymphocyte proliferation.

CONCLUSIONHumoral and cytokine responses elicited by pcDNA3.1 + S inoculation can be modulated by co-inoculation with pcIL-12 and efficiently induced Th1-dominant immune responses.

Animals ; Cytokines ; biosynthesis ; Genetic Therapy ; Hantavirus ; immunology ; Immunoglobulin G ; blood ; Immunophenotyping ; Interleukin-12 ; genetics ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid ; genetics ; immunology ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

Objective We aimed to assess the association between urinary bisphenol A(BPA)concentrations and gestational age in pregnant women. Methods A total of 248 pregnant women were recruited from a maternal and child care hospital in Shanghai. A questionnaire survey was completed to collect socio-demographic information and spot urine samples were collected during pregnancy. Gas chromatography-tandem mass spectrometry(GC-MS/MS)was used to measure BPA concentrations in urine samples. Linear relationship between urinary BPA level and gestational age was assessed by using generalized additive models. Multivariate regression model was used to evaluate associations of prenatal BPA exposure with gestational age. Results BPA was detected in all the urine samples. Median value and geometric mean of urinary BPA levels were 0.85 μg/L and 1.21 μg/L, respectively. Linear relationship between urinary BPA concentration and gestational weeks was confirmed(non-linear P > 0.05). Positive association between urinary BPA level and gestational age was indicated(regression coefficient, β = 0.19;95%CI:0.04-0.35;P = 0.016). However, it was only observed in girls, stratified by sex of newborns(β = 0.18;95%CI:0.03-0.34;P = 0.020). After stratification by trimester, no significant association was found in the second or the third trimesters. Conclusion Pregnant women are extensively exposed to BPA. Urinary BPA exposure during pregnancy may extend gestational age, especially in girls.

OBJECTIVETo investigate the restoration of erectile function by reconstructing cavernous nerves (CN) with small intestinal submucosa (SIS) grafts.

METHODSWe prepared SIS grafts, established rat models and divided the models into a CN ablation, a sham-operation and an SIS graft group. The CNs at both sides were severed with 1 cm ablated in the first group, and 0.5 cm removed in the third, followed by reconstruction with the SIS grafts. Three months after surgery, the apomorphine test was performed to evaluate the erectile function, and then all the rats were sacrificed to detect the expression of nNOS in the penis.

RESULTSPenile erection was observed in 72.73% (8/11) of the rats for (1.07 +/- 0.89) times within 30 min in the SIS graft group, as compared with 0% (0/11) of the rats for (0.00 +/- 0.00) times in the CN ablation group (P < 0.01), and 90.91% (10/11) of the rats for (2.19 +/- 1.17) times in the sham-operation group (P < 0.01). The number of nNOS nerve fibers was significantly larger in the SIS graft than in the CN ablation group (70.36 +/- 10.09 versus 22.09 +/- 4.76, P < 0.01), but both were significantly smaller than that of the sham-operation group (90.81 +/- 5.69, P < 0.01).

CONCLUSIONThe SIS grafting technique contributes to the recanalization of the severed CN and restoration of erectile function in rats after surgical injury.

Animals ; Erectile Dysfunction ; surgery ; Intestinal Mucosa ; transplantation ; Intestine, Small ; Male ; Nerve Regeneration ; Nerve Tissue ; injuries ; surgery ; Penile Erection ; Penis ; innervation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate status of infection with severe acute respiratory syndrome coronovirus (SARS-CoV) in traders of wild animals wholesale markets in Guangzhou.

METHODSSerum antibody against SARS-CoV IgG was determined cross-sectionally and symptoms of respiratory infection were investigated retrospectively for part of traders of three wholesale markets for wild animals in Guangzhou.

RESULTSOverall rate of infection with SARS-CoV in 635 traders was 16.69%, varying in three different markets. Infection rate in market A mainly engaging in wild animals ranked the highest of 25.61%, significantly higher than that in markets B and C engaging in domestic fowls and snakes. Infection rate in traders only engaging in civet cats was 58.54%, significantly higher than that in traders engaging in snakes only (9.46%). In market A, infection rate varied in different persons, 59.34%, 20.59%, 16.00%, 15.22%, 10.40% and 9.68% in traders engaging in wild animals, managers, children of the traders, traders engaging in domestic fowls, traders engaging in snakes, and traders engaging in frozen food, respectively, in a decreasing pattern as their contact opportunities. During the period of SARS epidemic, detection rate of SARS-CoV antibody in people with symptoms of acute respiratory infection was higher (30.70%) than that in those without such symptoms (20.08%). Prevalence of symptoms of acute upper respiratory infection in people with positive antibody against SARS-CoV was higher (49.28%) than that in those with negative antibody (30.35%).

CONCLUSIONSInfection with SARS-CoV in traders of animal markets possibly related to their direct exposure to wild animals, particularly to civet cats. During the period of SARS epidemic, some of the traders did infect with SARS-CoV, but they were neglected due to clinically inapparent manifestations.

Animals ; Antibodies, Viral ; blood ; China ; Contact Tracing ; Family ; Humans ; Immunoglobulin G ; blood ; Occupational Exposure ; Occupations ; classification ; Retrospective Studies ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; transmission

OBJECTIVEEstablishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy.

METHODSThe whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential display-PCR (RFDD-PCR). Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach.

RESULTSA total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, beta-arrestin, Phosducinìrod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mGluRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged.

CONCLUSIONThese results prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro-dysfunctions of diabetic retina.

Animals ; Diabetes Mellitus, Experimental ; genetics ; Diabetic Retinopathy ; genetics ; Female ; Gene Expression Profiling ; methods ; Rats ; Rats, Sprague-Dawley ; Retina ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo study the mechanism and clinical effect of using the manipulatin of move, adduction, rotation method for the treatment of shoulder dislocation.

METHODSfrom January 2010 to March 2012,120 patients with shoulder dislocation admitted were randomly divided into treatment group and control group, 60 cases in each group. In the treatment group, there were 31 males and 29 females; In control group, there were 30 males and 30 females. In treatment group, 60 patients create their own "on the move, adduction, rotation manipulative treatment,the other 60 cases in control group used traditional restoration methods in wearing the traction treatment. The efficacy of two methods of treatment of shoulder dislocation were compared on reset process once successfully rate, the patient's level of pain (VAS).

RESULTSIn treatment group, restoration once successfully were in 59 cases, improvement of the pain (VAS) values was 3.76 +/- 1.05, the results were excellent in 57 cases, good in 2, poor in 1,without concurrent fractures and nerve injury, without re-dislocation after 3 months. In control group, restoration once successfully were in 50 cases, improvement of pain (VAS) values was 5.67 +/- 1.15, the results were excellent in 45 cases, good in 7, poor in 8. The reset once successfully rate,improvement of VAS values and clinical effect in treatment group were better than that of control group.

CONCLUSIONOn the move, adduction, rotation method has advantages of more wearing traction with uniform force, work together to focus, saving time and effort, easy and convenient to avoid iatrogenic injury.

Adult ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Rotation ; Shoulder Dislocation ; therapy

Objective To conduct serological investigation on H5N1/H9N2/H7N7 infection among people occupied in poultry fields. Methods Serum samples were collected from people working in live poultry and none-poultry retailing food markets, poultry wholesaling, large-scale poultry breading factories and in small-scale farms, wide birds breeding, swine slaughtering houses and from normal population. Antibodies of H5, H9 and H7 with hemagglutination inhibition and neutralization tests were tested and analyzed. Logistic regression and χ2 test were used. Results Among 2881 samples, 4 were positive to H5-Ab(0.14%), 146 were positive to H9-Ab (5.07%) and the prevalence of H9 among people from live poultry retailing (14.96%) was the highest. Prevalence rates of H9 were as follows: 8.90% in people working in the large-scale poultry breading factories, 6.69% in the live poultry wholesaling business, 3.75% in the wide birds breeding, 2.40% in the swine slaughtering, 2.21% in the non-poultry retailing, 1.77% in the rural poultry farmers and 2.30% in normal population. None was positive to H7-Ab among 1926 poultry workers. Conclusion The H5 prevalence among people was much lower than expected, but the H9 prevalence was higher. None of the populations tested was found positive to H7-Ab. There was a higher risk of AIV infection in live poultry retailing, wholesaling and large-scale breading businesses, with the risk of live poultry retailing the highest. The longer the service length was, the higher the risk existed.

OBJECTIVETo investigate the possible risk factors of severe acute respiratory syndromes coronavirus (SARS-CoV) infection in workers from animal markets.

METHODSSelf-designed questionnaires were used and serum samples were tested. Logistic regression analysis was used to analyze the data.

RESULTSResults from simple factor logistic regression analysis showed that jobs which dealing with domestic livestock, wild livestock, wild animals, aquatics were related to risk factors of SARS-CoV infection. Results from multifactor logistic regression analysis showed that jobs that dealing with wild livestock and poultry were important risk factors with OR 12.28 and 0.41.

CONCLUSIONJob that dealing with palm civets was the main risk factor of SARS-CoV infection in animal market workers.

Animal Husbandry ; Animals ; Carrier State ; epidemiology ; virology ; China ; epidemiology ; Disease Reservoirs ; Humans ; Logistic Models ; Occupational Exposure ; Poultry ; Risk Factors ; SARS Virus ; immunology ; isolation & purification ; Severe Acute Respiratory Syndrome ; epidemiology ; virology ; Surveys and Questionnaires

OBJECTIVETo investigate the relationship between tea consumption, biliary tract cancers and gallstone disease.

METHODSA population-based case-control study was conducted in urban Shanghai from 1 June 1997 to 31 May 2001 involving interviews with 627 new cases of biliary tract cancers (including 368 cases of gallbladder cancer, 191 cases of extrahepatic bile duct cancer and 68 cases of cancer of the ampulla of Vater) aged 35 to 74 years and 959 population controls frequency-matched to cases by gender and age in five-year group. 1037 patients of gallstone disease were selected from the same hospital. All subjects were interviewed in person by trained interviewers by use of a structured questionnaire. Unconditional logistic regression analysis was used to calculate adjusted odds ratio (OR) and 95% confidence interval (CI).

RESULTSCompared with tea non-drinkers, current tea consumption was inversely associated with risk of gallbladder cancer, extrahepatic bile duct cancer and gallstone disease among females with OR of 0.57 (95% CI: 0.34-0.96), 0.53 (95% CI: 0.27-1.03) and 0.71 (95% CI: 0.51-0.99), respectively. OR declined with younger age at initiation of tea drinking and with longer duration of tea consumption (P for trend < 0.05). Among males, the corresponding OR were mostly below one, although not statistically significant.

CONCLUSIONTea consumption may decrease the risk of cancers of the gallbladder and extrahepatic bile duct among females. The protective effect appears to be independent of gallstone disease.

Adult ; Aged ; Bile Ducts, Extrahepatic ; Biliary Tract Neoplasms ; epidemiology ; etiology ; Case-Control Studies ; China ; epidemiology ; Female ; Flavonoids ; pharmacology ; Gallbladder Neoplasms ; epidemiology ; etiology ; Gallstones ; epidemiology ; etiology ; Humans ; Male ; Middle Aged ; Phenols ; pharmacology ; Polyphenols ; Protective Agents ; pharmacology ; Risk Factors ; Tea ; chemistry