A high active soybean isoflavone glucoside hydrolase-producing mould strain was isolated from spirit qu. Its optimal hydrolase-producing conditions were as follows: 2.5% wheat bran as carbon source, 1% NaNO3 as nitrogen source, initial pH7. 0, culture medium volume 40mL/250mL, inoculating quantity 8% , culture temperature 30℃, revolutions 160r/min and culture time 84h. The enzyme activity reached 82 U/mL. Cu2+ can inhibit Absidia sp. R strain from producing the hydrolase, the influence of other metal ions was not remarkable on it.

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.

OBJECTIVETo study the effects of early high fat diet on sugar metaboliam, insulin sensibility and pancreatic β cellularity in young rats.

METHODSSixty male weaned young rats were randomly fed with high fat diet (high fat group) and normal diet (control group). The body weight, viscus fattiness and fasting plasma glucose (FPG) were measured after 3, 6 and 9 weeks. Serum insulin level was measured with radioimmunoassay. The ultrastructure of pancreas was observed under an electricmicroscope.

RESULTSThe high fat group had significantly higher body weight and visceral fat weight than the control group after 3 weeks. There were no significant differences in the FPG level between the two groups at all time points. The levels of fasting insulin and HOMAIR in the high fat group were significantly higher than those in the control group after 3, 6 and 9 weeks (P<0.01). Dilation of rough endoplasmic reticulum and mild swelling of mitochondria of islet β-cells were observed in the high fat group after 6 weeks.

CONCLUSIONSEarly high fat diet may induce a reduction in insulin sensitivity and produce insulin resistance in young rats. Endoplasmic reticulum expansion in β-cells may be an early sign of β-cell damage due to obesity.

Animals ; Blood Glucose ; analysis ; Dietary Fats ; adverse effects ; Insulin ; Insulin Resistance ; Insulin-Secreting Cells ; pathology ; ultrastructure ; Intra-Abdominal Fat ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the changes of nerve conduction velocity in degenerative rabbit sciatic nerve authograft induced by 60Co irradiation after orthotopic replantation. Methods A 30-mm-long segment was severed from normal adult rabbit sciatic nerve and exposed to 60Co irradiation at the dose of 350 Cry to induce neural degeneration. The nerve segment was then replanted orthotopicaily, and the nerve conduction velocity was determined using electrophysiological test at 4, 6 and 8 months after the replantation. Results At 6, 8 months after the replantation, the nerve conduction velocity in the degenerative nerve autograft showed no significant difference from that in normal sciatic nerve (P>0.05). But at 4 months after the replantation, the nerve conduction velocity in the autograft was significantly lower than the normal velocity (P<0.05). Conclusion The nerve conduction velocity can be obtained by replantation of a long (3 mm) degenerative nerve segment due to 60Co irradiation.

Objective To detect the effect and role of profound hypothermia on cytochrome C release and cell apoptosis in the hippocampus after global ischemia in rats. Methods We established the animal model of cardiopulmonary bypass (CPB) in 24 rats which were randomized into 3 even groups:blank control group,normothermia ischemia group and hypothermia ischemia group.After the hippocampus samples were harvested from the 3 groups,Western blot was used to detect the expression of cytochrome C (Cyt-C) and TUNEL was used to measure the cell apoptosis. Results The expression of Cyt-C in the normothermia ischemia group increased significantly compared with the blank control group but decreased significantly compared with the hypothermia ischemia group (P＜0.05).The rate of cell apoptosis in the normothermia ischemia group increased significantly compared with the blank control group but decreased significantly compared with the hypothermia ischemia group (P＜0.05).Conclusions Profound hypothermia can inhibit cytochrome C release into the cytoplasm and thus reduce cell apoptosis in the hippocampus after global ischemia in rats. This may be one important mechanism underlying the protective effect of profound hypothermia.

This study was aimed to analyze the BP1 binding site sequence upstream of β-globin gene in Chinese Han population, and to investigate polymorphism in the BP1 binding site upstream of β-globin gene, so as to provide the basis for exploration of relation between polymorphisms in the BP1 binding site and β-globin expression. Genomic DNA was extracted from peripheral leukocytes of 110 healthy individuals in Chinese Han population. Sequence of the BP1 binding site upstream of β-globin gene was amplified by polymerase chain reaction, the polymorphic variation in the BP1 binding site was determined by DNA sequencing. The results indicated that 2 polymorphism loci were found in the BP1 binding site upstream of β-globin gene, they were C/T at the -551 bp region and (AC)(n)(AT)(x)T(y) at the -530 bp region in Chinese Han population. Frequencies of C and T were 60.4% and 39.6% at position -551. Analysis of the (AC)(n)(AT)(x)T(y) polymorphism revealed 9 different genotypes: (AC)(2)(AT)(9)T(5), (AC)(2)(AT)(8)T(5), (AC)(2)(AT)(7)T(7), (AC)(3)(AT)(7)T(5), (AC)(2)(AT)(8)T(9), (AC)(3)(AT)(8)T(5), (AC)(2)(AT)(10)T(3), (AC)(2)(AT)(11)T(3), and (AC)(2)(AT)(7)T(5) at position -530. Frequencies of 9 (AC)(n)(AT)(x)T(y) polymorphisms were 33.2%, 29.1%, 24.1%, 5.4%, 3.2%, 1.8%, 1.4%, 0.9%, and 0.9% respectively. It is concluded that rich (AC)(n)(AT)(x)T(y) polymorphisms at the -530 bp region in the BP1 binding site upstream of β-globin gene are found in Chinese Han population. (AC)(2)(AT)(9)T(5), (AC)(2)(AT)(8)T(5), and (AC)(2)(AT)(7)T(7) are 3 major polymorphisms among Chinese Han population, and (AC)(3)(AT)(8)T(5) is a novel polymorphism at the -530 bp region. More studies should be done to explore relation between (AC)(n)(AT)(x)T(y) polymorphisms and β-globin expression.
Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Binding Sites ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymorphism, Genetic ; beta-Globins ; genetics

OBJECTIVETo observe anti-HEV IgG response to vaccination of recombinant antigen fragments and evaluate its protection from Hepatitis E Virus infection in rhesus monkeys (Macaca mulatta).

METHODSTwelve monkeys were divided into three groups and immunized respectively with three different recombinant antigens: namely Ag1 (carboxyl terminal 431 amino acids of ORF2), Ag2 (128aa fragment at the carboxyl terminal of ORF2), and Ag3 (full length ORF3 ligated with two ORF2 fragments encoded by 6743-7126nt and 6287-6404nt). The monkeys were challenged intravenously with fecal suspension from experimentally infected rhesus monkeys, and the other three monkeys served as the placebo group for challenge with HEV. The dynamic changes of the levels of ALT and anti-HEV IgG were examined. Pathological changes of liver tissue were observed by light microscope. Excretion of virus was detected by RT-nPCR.

RESULTSHepatic histopathology of two monkeys in the placebo group was consistent with acute viral hepatitis, and ALT was elevated 3-4 weeks after inoculated with virus, up to 10-20 times higher than normal level. The liver tissue of monkeys immunized with antigen kept normal, ALT in several monkeys elevated mildly, and anti-HEV IgG conversation occurred at 1-2 weeks after vaccination, with the titer reaching 1:12,800. The virus RNA could be detected by RT-nPCR from days 7 to 50 in monkeys of control group, and from days 7 to 21 in vaccinated monkeys after challenged with virus.

CONCLUSIONSThe recombinant antigens could induce the production of anti-HEV IgG, which protected rhesus monkeys from acute Hepatitis symptoms related to HEV infection.

Animals ; Antigens, Viral ; immunology ; Hepatitis E ; prevention & control ; Hepatitis E virus ; immunology ; Immunoglobulin G ; immunology ; Macaca mulatta ; RNA, Viral ; blood ; Recombinant Proteins ; immunology ; Vaccination ; Viral Hepatitis Vaccines ; immunology

OBJECTIVETo study the variation of specific antibody among convalescent of severe acute respiratory syndrome (SARS) patients through a three-year program.

METHODSSera samples were collected from SARS cases in the 5th, 20th and 35th month after onset of the illness. The SARS-CoV specific antibody was detected for all of them by ELISA and neutralized test simultaneously. The titer of neutralizing antibodies was calculated using Reed-Muench method, and the comparison between different time groups was analyzed regarding the variance of data on repeated measures after logarithm conversion.

RESULTS13, 17 and 13 sera samples were collected in the 5th, 20th and 35th month after onset. Results showed that despite the fact that the positive rates of ELISA antibody were 100%, 82.4% and 84.6% respectively,the neutralizing antibody was still positive for all the samples. The average neutralizing antibody titers were 1:43 (1:16-1:203), 1:36 (1:17-1:59) and 1:21 (1:10-1:39) on the 5th, 20th and 35th month after onset, and the differences were statistically significant (F = 60.419, P < 0.001). On the 35th month after the onset, 30.8% (4/13) of the patients were still having the neutralizing antibody level of above 1:36, but the neutralizing antibody level in another 30.8% (4/13) of the patients had decreased to as low as 1:10, when the cut-off level was set as 1:8.

CONCLUSIONResults of the study indicated that the neutralizing antibody of SARS cases could last for at least three years, but the sera specific antibody in SARS cases decreased gradually when time went by. However, neutralizing antibody in some of the cases decreased to a lower level on the 35th month. Further follow-up study was worthwhile to observe the long-lasting profile of antibody existence on SARS cases.

Antibodies, Neutralizing ; analysis ; Enzyme-Linked Immunosorbent Assay ; Follow-Up Studies ; Humans ; Severe Acute Respiratory Syndrome ; immunology

OBJECTIVETo investigate the effects of anluohuaqianwan on experimental hepatic fibrosis induced by dimethyl nitrosamine (DMN) in rats.

METHODS36 male SD rats were randomly dividied into three groups: model group, normal group, anluohuaqianwan group. The rats in the three groups were treated with DMN daily for 4 weeks. The liver function was detected using auto biochemistry analyzer, the serum HA, LN, IV-C, PIIIP were detected by immunoradiometry, the histopathology was observed in the left liver lobe after HE staining, the expression of matrix metalloproteinase-2 (MMP-2) in liver tissue were detected by immunohistochemistry.

RESULTSThe serum levels of ALT, AST, ALP, TP, ALB and the contents of HA, LN, IV-C in model group were significantly increased compared to these in the normal group (P less than 0.01). The serum levels of ALT, AST and the contents of HA in anluohuaqianwan group were significantly lower than those in the model group (P less than 0.01). The liver MMP-2 in the model group was significantly increased compared to that in the normal group (P less than 0.05). The expression of MMP-2 in liver tissue of model group was lower than that in the anluohuaqianwan group (P less than 0.05).

CONCLUSIONAnluohuaqianwan can inhibit liver fibrosis in rats induced by DMN.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Dimethylnitrosamine ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hyaluronic Acid ; blood ; Hydroxyproline ; metabolism ; Immunohistochemistry ; Liver ; drug effects ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; metabolism ; pathology ; Liver Function Tests ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the natural process of adenoid growth and degeneration as the age grows, to investigate the related clinical significance and pathologic characteristics of hypertrophied adenoid.

METHODSTotally 2650 (age 2 to 87) cases with nasal obstruction or/and other symptoms were included in the patients group, and 810 (age 3 to 85) subjects without symptoms were included as the control group. Morphological characteristics examined with nasal endoscope. Biopsy was performed for 39 cases. The adenoid was calcified as 4 degrees according to the size.

RESULTSIn the patient group, age 2 to 9, degree III and degree II adenoid were 81.1% (198/244) and 18.9% (46/244) respectively. And adenoid of children whose age 2 to 5 was 100.0% in degree III; In above 10 years old group, the adenoid was mostly degree II. In age 60 to 69 group, degree 0 was (66.5%), and in age 81 or above, degree 0 reaches 100%. And 19 years old was the youngest age at which adenoid of degree 0 started to be found and 21 was the oldest age at which there is no adenoid of degree III. In the control group, compared with the patient group, no statistical significant difference found in all other groups except in age 2 to 9 (degree III 57.9%, 22/38, degree II 42.1%, 16/38). Shapes of adenoids at degree II varied while degree I were almost like peeled orange. Pathologically, among children there are abundant of adenoidal lymph tissue, while in adults the lymph tissue getting less as age grows but with evident inflammation reaction. Among patients, the incidence of sinusitis and snoring was higher in degree III group compared with others, 47.4% and 18.7% respectively, and the differences is statistically significant (chi2 = 51.28, P < 0.01; chi2 = 40.26, P < 0.01).

CONCLUSIONSAdenoid volume of children (age < 10) is the biggest, especially of children under 5 years old.

Adenoids ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Child ; Child, Preschool ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Nasal Obstruction ; pathology ; surgery ; Young Adult