Objective To study the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in preeclampsia placenta and the relation with preeclampsia attacks.Methods Forty-four samples from pregnant women with preeclampsia (preeclampsia group),38 samples from pregnant women with eclampsia,and 49 samples from normal pregnancies (control group) were obtained.We detected the expression of EMMPRIN in placenta by immunohistochemistry and the expression of EMMPRIN mRNA by RT-PCR,Results (1) EMMPRIN positive expression:in preeclampsia group,the moderate expression rate was 18% (8/44) and the strong positive rate was 9% (4/44);in eclampsia group moderate positive rate was 21% (8/38) and strong positive rate 13% (5/38).The difference of the two groups was insignificant (P＞0.05).In control group the moderate positive rate was 12% (6/49) and strong positive rate 82% (40/49),the difference from the preeclampsia and the eclampsia groups was significant (P＜0.001).(2)EMMPRIN mRNA expression:in preeclampsia group EMMPRIN mRNA expression in term placenta (37-40 gestational weeks) was 0.342?0.002,and in eclampsia group 0.344?0.023;the difference between the two groups was insignificant (P＞0.05).In control group EMMPRIN mRNA expression in term placenta (37-40 gestational weeks) was 0.872?0.094,the differences between the control group and preeclampsia and eclampsia groups were both significant (P＜0.001).Conclusion The decrease in the expression of EMMPRIN in placenta is an important cause of preeclampsia onset;expression rate of EMMPRIN may serve as an indicator in predicting preeclampsia.

OBJECTIVETo study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection.

METHODSFab monoclonal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of V(H) and V(L) and neutralizing activity on HCMV AD169 stain were analyzed in vitro. The light and heavy chain Fd fragment genes of Fab antibodies were further cloned into a recombinant baculovirus expression vector pAC-kappa-Fc to express intact IgG. Secreted products were purified with affinity chromatography using protein G.

RESULTSSDS-PAGE and Western blot confirmed the expression of the intact IgG. Immuno-blotting and -precipitation were used to identify HCMV proteins. One Fab monoclonal antibody recognized a conformational HCMV protein.

CONCLUSIONIgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 microg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans.

Amino Acid Sequence ; Animals ; Base Sequence ; Cell Line ; Cloning, Molecular ; Cytomegalovirus ; immunology ; DNA, Viral ; genetics ; Genes, Viral ; Humans ; Immunoblotting ; Immunoglobulin G ; immunology ; Immunoprecipitation ; Insecta ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; immunology

Objective To discuss the effects of different exercise frequency and diet guidance for fat loss on obese patients, and to develop a safe and effective multidisciplinary intervention program for obese patients. Methods A total of 140 members of Ningbo Slimming Club were recruited, with the age between 18-60 years, body mass index (BMI) greater than 28 kg/m2, and they were randomly assigned to low frequency group and high frequency group. In low frequency group, the intervention was complete 100 minutes aerobic exercise and 3 times a week. In high frequency group, the intervention was complete 50 minutes of aerobic exercise and 6 times a week. Each group followed the same diet weight loss program. By comparing each index before and after the intervention, the effect of fat loss was evaluated. Results After intervention, systolic blood pressure, diastolic blood pressure, body weight, BMI, waist circumference, body fat percentage, triglyceride, low density lipoprotein and total cholesterol in two groups were decreased, and high density lipoprotein were increased (P<0.05) . Compared between the low frequency and high frequency exercise group, there were no significant difference for the systolic blood pressure, diastolic blood pressure, body weight, waist circumference, BMI, triglyceride, high density lipoprotein, low density lipoprotein and total cholesterol levels (P>0.05), but body fat percentage in low frequency exercise group decreased significantly (P< 0.05) . Conclusion Diet intervention combined with exercise has a significant effect of fat loss on obese patients. Based on the same weekly aerobic exercise time, obese patients with low frequency of exercise had better fat loss than obese patients with high frequency of exercise.

OBJECTIVETo measure the range of plasma hydrogen sulfide (H2S) in children.

METHODSTotally 200 healthy children were classified into 4 groups based on age and sex: 7-14 years old group (n = 75, 43 boys and 32 girls), 15-19 years old group (n = 125, 64 boys and 61 girls). Plasma H2S level was detected by a modified sulfide electrode-based method.

RESULTSPlasma H2 S levels were (52.2181 +/- 17.9400) micromol/L in 7-14 years old boys, (51.9441 +/- 16.5448) micromol/L in 7-14 years old girls, (52.8771 +/- 14.1444) micromol/L in 15-19 years old boys, and (53.6551 +/- 14.5563) micromol/L in 15-19 years old girls (P > 0.05). In summary, the range of plasma H2S in children was about (52.8234 +/- 15.4339) micromol/L.

CONCLUSIONThe range of plasma H2S in children is about (52.8234 +/- 15.4339) micromol/L.

Adolescent ; Age Factors ; Blood Gas Analysis ; methods ; Child ; Female ; Humans ; Hydrogen Sulfide ; blood ; Male ; Reference Values ; Sex Factors

BACKGROUNDCloning recombinant human Fab fragment against HCMV for the purpose of prophylaxis and control of HCMV infection.

METHODSThe authors constructed a HCMV phage display library with 2 x 10(6) clones, then used purified HCMV viral lysates to pan the library, then screened by ELISA.

RESULTSThree clones showed positive responses in ELISA, they also showed high specificity in IFA, two of them could neutralize HCMV in neutralizing assays.

CONCLUSIONThe specific binding of Fab antibodies to HCMV was demonstrated by ELISA, IFA and neutralizing activities. These results provide us the basis for further research of neutralizing recombinant human whole IgG molecule.

Antibodies, Viral ; genetics ; immunology ; Cytomegalovirus ; genetics ; immunology ; Cytomegalovirus Infections ; immunology ; virology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Neutralization Tests ; Peptide Library

OBJECTIVETo investigate the expression of the lysosomal enzyme acid sphingomyelinase (ASMase) in alcohol-induced hepatic fibrosis using a rat model.

METHODSThe model of liver fibrosis was induced by administration of alcohol and high fat diet using 20 rats. Six rats given no alcohol and normal diet served as the control group. Real-time PCR, western blotting, and immunohistochemistry were used to evaluate fibrosis-related changes in the mRNA and protein expressions of ASMase.

RESULTSThe fibrotic liver tissues of the model rats showed significantly higher expression levels of ASMase than the non-fibrotic liver tissues of the control rats (P less than 0.05).

CONCLUSIONExpression of ASMase is increased in the fibrotic liver tissue of an alcohol-induced hepatic fibrosis rat model, suggesting that this lysosomal enzyme may contribute to development of this disease condition.

Animals ; Liver ; enzymology ; Liver Cirrhosis, Alcoholic ; enzymology ; Liver Cirrhosis, Experimental ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley ; Sphingomyelin Phosphodiesterase ; metabolism

OBJECTIVETo study the influence of acute pancreatitis in pregnancy (APIP) on pregnancy outcomes and neonates.

METHODSA retrospective analysis was performed for 33 APIP patients and 31 neonates born alive.

RESULTSOf the 33 APIP patients, 26 (79%) developed APIP in the late pregnancy. Fourteen (45%) patients had hyperlipidemic APIP, 13 (42%) had biliary APIP, and 4 (13%) had other types of APIP. According to the severity, 22 (67%) were mild APIP, 5 (15%) were moderate APIP, and 6 were severe APIP. None of the 33 APIP patients died. Among the 20 patients with term delivery, 11 underwent termination of pregnancy; among the 10 patients with preterm delivery, 9 underwent termination of pregnancy; two patients experienced intrauterine fetal death, and one experienced abortion during the second trimester. Among the 31 neonates born alive (two of them were twins), 1 (3%) died, 12 (39%) experienced neonatal hyperbilirubinemia, 8 (26%) had neonatal hypoglycemia, 6 (19%) had neonatal respiratory distress syndrome, 5 (16%) experienced infectious diseases, and 2 (6%) experienced intracranial hemorrhage. The hyperlipidemic APIP group had a higher percentage of patients undergoing termination of pregnancy than the biliary APIP and other types of APIP groups (P<0.05). The incidence rate of preterm infants in the moderate APIP was higher than in the mild and severe APIP groups (P<0.05). The mean birth weights of neonates were the lowest in the moderate APIP group. The incidence rates of neonatal respiratory distress syndrome, intracranial hemorrhage, and infectious disease were the lowest in the mild APIP group (P<0.05).

CONCLUSIONSAPIP can lead to adverse pregnancy outcomes and neonatal diseases, which are associated with the severity of pancreatitis.

Acute Disease ; Birth Weight ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Pancreatitis ; complications ; Pregnancy ; Pregnancy Complications ; Retrospective Studies

OBJECTIVETo study the effect of rosiglitazone on atherosclerosis and potential mechanism in ApoE-knockout mice.

METHODSThirty-two 6-week-old ApoE-knockout mice were used as atherosclerosis model in two groups: rosiglitazone group (n = 18) and control group (n = 14). Each group contained equal numbers of male and female mice. All mice were fed with normal chow diet. In addition to normal diet, rosiglitazone group received rosiglitazone 17 mg/kg of body weight/day. Venous bloods were collected for plasma glucose and lipid analysis, and aorta were prepared for morphologic and immunohistochemical analysis after 14 weeks. Aortic root (1 cm) was cut and prepared for paraffin slice. The histomorphometric analysis of atherosclerotic lesion was performed by means of HE; positive percentage of macrophage cell and tumor necrosis factor-alpha were measured by means of immunohistochemistry in cross section. The ratio of lesion/aortic wall surface in the rest aorta was measured by means of Sudan IV staining in longitudinal section.

RESULTSThe amount of fatty streak in rosiglitazone group was significantly greater than that of control group; the gross number of lesions and the number of fibrous plaque and atheromatous plaque were similar in two groups. There were no differences in percentage of lesions in cross section in two groups. Rosiglitazone could significantly reduce the extend of atherosclerosis of longitudinal section, decrease the amount of macrophage cell and the level of tumor necrosis factor-alpha in lesions. The plasma glucose was normal and similar in two groups, and total cholesterol, LDL-cholesterol and triglyceride were significantly higher in rosiglitazone group.

CONCLUSIONRosiglitazone suppresses the expression of tumor necrosis factor-alpha, reduces the number of macrophage cell in lesion, and inhibits the development of atherosclerosis.

Animals ; Aorta ; pathology ; Apolipoproteins E ; genetics ; physiology ; Atherosclerosis ; blood ; pathology ; prevention & control ; Blood Glucose ; analysis ; Body Weight ; drug effects ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Hypoglycemic Agents ; therapeutic use ; Macrophages ; pathology ; Male ; Mice ; Mice, Knockout ; Thiazolidinediones ; therapeutic use ; Tumor Necrosis Factor-alpha ; biosynthesis

OBJECTIVETo identify genes in human cells infected with high pathogenic avian influenza viruses H5N1.

METHODSThe lung carcinoma cells line A549 was infected with H5N1 and H1N1, respectively. We harvested the infected cells at the different time points after infection and screened the genes with differential expression via microarray technology. The candidate genes were selected and confirmed by quantitative real-time PCR.

RESULTSThe spectrum of genes with the differential expression in the cells infected with H5N1 was obtained and 16 candidate genes were identified in the cellular apoptosis pathway, mTOR pathway, and the cellular immunity as well.

CONCLUSIONSOur results suggest that H5N1 exert a stronger impact on eliciting apoptosis of infected cells than the common influenza virus H1N1.

Animals ; Apoptosis ; Cell Line, Tumor ; Gene Expression Profiling ; Humans ; Influenza A Virus, H1N1 Subtype ; physiology ; Influenza A Virus, H5N1 Subtype ; physiology ; Influenza, Human ; genetics ; metabolism ; virology ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection.

METHODSBy using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined.

RESULTSAfter four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus.

CONCLUSIONThe SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection.

Amino Acid Sequence ; Animals ; Antibodies, Viral ; immunology ; Cercopithecus aethiops ; Humans ; Membrane Glycoproteins ; immunology ; Neutralization Tests ; Peptide Library ; Protein Binding ; Protein Engineering ; Recombinant Proteins ; immunology ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; virology ; Spike Glycoprotein, Coronavirus ; Vero Cells ; Viral Envelope Proteins ; immunology ; Viral Matrix Proteins ; immunology