Objective: To analyze the recombination of full-length genomic sequences of novel influenza virus A/H1N1 in 2009 pandemic. Methods: The full-length sequences of the novel A/H1N1 and reference sequences were downloaded from NCBI database. MEGA4.0 software was used to connect, align sequences, and analyze the similarity between the full-length sequences of the novel virus and each of the reference strains. Recombination was analyzed by Simplot software (version 3.5.1). Results: Simplot analysis indicated that the PB1 genes (polymerase B1, PB1) of the novel A/H1N1 viruses might evolve from human H3N2 virus (identity: 93.7%); the PB2 genes (polymerase B2, PB2) and the PA genes (polymerase A, PA) might evolve from avian H5N1 viruses (identity: 89.0%, 89.9%, respectively); the HA genes (hemagglutinin, HA), the NP genes (nucleoprotein, NP) and the NS genes (non-structural protein, NS) showed high similarities with those of swine H1N1 viruses isolated in North America (identity: 91.7%, 93.1%, and 93.1%, respectively); and the NA genes (neuraminidase, NA) and the MP genes (matrix protein, MP) might evolve from European swine H1N1 viruses (identity: 90.5%, 95.5%, respectively). The full-length sequence of the novel A/H1N1 viruses had a highest similarities with swine H1N1 viruses isolated in North America (identity: 83.9%). Conclusion: The novel influenza virus A/H1N1 is a recombinant virus evolving from human H3N2 viruses, swine H1N1 from North America, swine H1N1 from Europe, and swine H5N1 from Asia.

Objective: To investigate the role of HBV subgenotypes B2, C2 in the carcinogenesis, treatment and prognosis of hepatocellular carcinoma (HCC). Methods: HBV genotypes and subgenotypes were detected in 462 HCC patients and 234 chronic hepatitis B (CHB) patients by a multiplex PCR assay, and HCC patients infected with HBV B2 or C2 were followed up for a year after surgical resection, transarterial chemoembolization(TACE) or a combination of both. Results: The HCC patients infected with HBV C2 had a higher chance to receive surgical treatment than those with B2 (P=0.007). Age of 40 years or older (P=0.030), male gender (P= 0.000), and viral load (>10 000 copies/ml) (P=0.017) were the independent risk factors for the carcinoge-nesis of HCC by using multivariate logistic analysis; however, there was no significant difference in the carcinogenesis of HCC between CHB patients with HBV subgenotypes B2 and C2. Age of 50 years or younger (P=0.044), infection with HBV B2 (P=0.027), and non-surgical treatment (P=0.000) were the independent risk factors for the recurrence of HCC. Thick trabecular type was more prevalent in HCC patients infected with HBV B2, C2 and genotype mixture (85.7%, 71.2% and 75.0%, respectively), and the proportions of histopathological types were not significantly different between HCC patients infected with HBV B2, C2 and genotype mixture. HBV subgenotype C2 was found in all HCC patients with rare histopathological type and subgenotype B2 and mixture were no found. Conclusion: There is no significant difference in the carcinogenesis of HCC between CHB patients with HBV subgenotypes B2 and C2. The HCC patients infected with HBV B2 have a lower chance to receive surgical treatment and are more severe than those with C2. HBV B2 is also closely associated with recurrence of HCC.

Objective: To elucidate the distribution of HBV genotypes and subgenotypes in patients with chronic hepatitis B (CHB), hepatocellular carcinoma(HCC) and asymptomatic HBV carriers(ASC) in Shanghai and areas around Shanghai, and to analyze the role of HBV genotypes and subgenotypes in the carcinogenesis and progress of HBV-related diseases. Methods: The HBV genotypes and subgenotypes were determined in 462 HCC patients, 234 CHB patients and 110 ASCs from Shanghai and areas around Shanghai by a multiplex PCR assay. Results: Genotypes A, B, C and D and subgenotypes B2, C1 and C2 were detected. Genotype C(mainly C2, 98.5%) and B(B2, 100%) were more prevalent than other genotypes in our group. Compared with CHB group, HCC group had higher proportion of genotype C(P=0.009) and lower proportion of genotype B(P=0.045). In the patients infected with HBV subgenotypes B2 or C2, the expression of HBeAg in CHB group was significantly higher than that in HCC group(P=0.005; P=0.008), and the expression of anti-HBe was lower in CHB group(P=0.003,P=0.001). In HCC patients, expression of HBeAg in patients infected with mixture genotype was higher than that in those infected with other genotypes(P=0.016 for B2). HCC patients (aged from 40 to 60) with HBV B2 infection had lower viral load than those with C2 and genotype mixture(P=0.029, P=0.021); and patients with HBV C2 infection had lower viral load than those with genotype mixture(P=0.041). Conclusion: Subgenotype C2 is more prevalent than B2 in people living in Shanghai and areas around Shanghai. The compositions of HBV genotypes and subgenotypes are different in patients with CHB, HCC and ASCs. Co-infection with different HBV-genotypes is associated with higher viral load, expression of HBeAg and easier carcinogenesis of HCC.

OBJECTIVETo investigate the effects of epigallocatechin gallate (EGCG)against exercise-induced fatigue in mice.

METHODSTotal 120 mice were randomly divided into three groups and tested separately. For each test, there were 30 mice subdivided into high dose (50 mg/kg . d EGCG) and low dose (10 mg/kg . d EGCG) groups as well as saline control group(1 ml/kg . d) with 10 in each. Burden swimming, running wheel endurance, stick climbing and hypoxia tolerance exercise were used to establish fatigue mice training model in three groups. And intraperitoneal injection with different doses of EGCG per day for consecutively 28 days and the mice in the control group were treated with normal saline. After the last each test, the blood lactic acid (BLA), blood urea nitrogen (BUN), blood lactate dehydrogenase (LDH), muscle glycogen (MG) and liver glycogen (LG) of each group of mice were determined.

RESULTSEGCG treatment groups(B and C)revealed a prolonged the mice survival time of burden swimming test, hypoxia tolerance, running wheel time and the ability of stick climbing(P < 0.05 or P <0.01), and increased LDH activity and MG and LG contents, reduced contents of BLA and BUN. High dose group had an obviously increase effect than lower dose group(P <0.05).

CONCLUSIONEGCG has significant effects against exercise-induced fatigue in mice.

Animals ; Blood Urea Nitrogen ; Catechin ; analogs & derivatives ; pharmacology ; Exercise Tolerance ; Fatigue ; drug therapy ; Glycogen ; metabolism ; L-Lactate Dehydrogenase ; blood ; Mice ; Physical Conditioning, Animal

Animals ; Catechin ; analogs & derivatives ; pharmacology ; Disease Models, Animal ; Fatigue ; metabolism ; physiopathology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

Aging ; Health ; Humans ; Longevity

Objective To investigate the value of mix recombinant antigen in schistosomiasis diagnosis. Methods The recombinant antigens of SjC23 (HD),SjC21.7 and SjCMP10 were expressed in vitro and purified by the affinity chromatography method. The efficacies of soluble egg antigen (SEA),single recombinant antigen and mix recombinant antigen for schistosomiasis diagnosis by Enzyme-linked Immunosorbent Assay were compared. Results The diagnostic efficacy was the same when the antibody IgG of the same group sera of schistosomiasis was detected by different quantities of 2.5 ?g/ml and 7.5 ?g/ml of SEA immobilized on microplate, and their absorbency A was the same, but there was a significant difference in the diagnostic efficacies between single recombinant antigen and mix recombinant antigen when the antibody IgG of the same group sera of schistosomiasis was detected by the same quantity of single recombinant antigen or mix recombinant antigen immobilized on microplate, the absorbency A of mix antigen reacted with the sera of schistosomiasis was significant higher than that of the single recombinant. The positive rates were very similar when 39 sera of acute schistosomiasis,80 sera of chronic schistosomiasis and 27 sera of advanced schistosomiasis were detected by SEA or mix recombinant antigen by ELISA in the same time. No cross-reaction presented when 20 clonorchiasis sera were detected by the mix recombinant antigen and no false positive presented when 40 of healthy sera were detected by the mix recombinant antigen. Conclusion The schistosomiasis diagnostic method by using the mix recombinant antigen has been established, which is helpful for improving the efficacy of schistosomiasis diagnosis.

Objective To synthesize and fusion express the predicted T-cell epitopes of Schistosoma japonicum, and analyze their immunogenicities. Methods The plus and minus oligo-nucleic acid strands of epitopes P7, P17, P18 were synthesized following their DNA sequence, respectively. The Nco I restriction enzyme sites were added to the 5′ end of epitope gene and the Xho I restriction enzyme sites were added to the 3′ end of epitope gene. The plus and minus strand of each epitope gene was annealed to form double strand DNA fragments. Then the double strand DNA fragments encoding epitope peptide were cloned into the site between Nco I and Xho I of plasmid pET32c(+) to construct recombinant plasmid which was transformed into E.coli DH5?. The recombinant plasmid containing P7, P17, P18 genes respectively was identified by PCR, restriction digestion and DNA sequencing, and then transformed into E.coli BL21 (DE3) for expressing the fusion protein. The fusion protein of peptide-thioredoxin(Trx) was expressed by inducing with IPTG and analyzed with SDS-PAGE. The fusion proteins were purified with Ni2+ column affinity chromatography. Meanwhile, the peptides P7, P17, P18 were synthesized artificially following their amino acid se-quence. By using the purified epitope peptide fusion proteins and synthesized epitope peptides, the splenic cells of C57BL/6J mice immunized with ultraviolet-attenuated cercaria of Schistosoma japonicum were stimulated respectively. The stimulation activity of fusion proteins and synthesized peptides were assayed by detecting the incorporation rate of 3 H-thymidine. Results The double strand DNA fragments of epitopes P7, P17, P18 were successfully cloned to form recombinant plasmids, all of which could express a 20 kDa fusion protein. Both the fusion protein and synthesized epitope peptides of P7 and P17 were able to stimulate the lymphocyte cells to proliferation effectively. Conclusion The peptide P7 and peptide P17 are testified as T-cell epitopes of Schistosoma japonicum.

Objective To explore the protective effect of recombinant superoxide dismutase(SOD)fusion protein against the infection of Schistosoma japonicum Chinese strain.Methods The recombinant SOD fusion protein was expressed and purified with Glutathione sepharose 4B.C57BL/6J mice were immunized with the recombinant SOD fusion protein mixed with Freund adjuvant.Four weeks after the final immunization,the mice of the experiment and control groups were challenged with(45?2)S.japonicum cercariae.All the mice were sacrificed on the forty-fifth day after the challenge to calculate the worm reduction rate and egg reduction rate,and to observe the pathologic changes of liver tissue of the mice.Results The worm reduction rate was 35.63% and the egg reduction rate was 31.17% in the experiment group.The number of granuloma in the live tissue of the experiment group was less than that of the control group,and the mean diameter of single granuloma in the experiment group reduced by 22.32% compared with that of the control group.The IgG subclass levels of IgG1,IgG2a,IgG2b were higher than those of the control group.Conclusion The recombinant SOD fusion protein has a protective effect against Schistosoma japonicum infection.

Objective To investigate the clinical significance of generalized epilepsy with febrile seizures plus(GEFS+ ). Methods The data of one family with GEFS+ were retrospectively analyzed by studying clinical manifestations, physical examinations, electroencephalogram(EEG), 24 hours dynamic EEG monitoring, et al. Some of the patients were examined by CT. Results Ⅳ 12, her chief complaints when admitted to hospital were frequent spasm for 3 days. She began to appear febrile seizures (FS) from 8 months after birth, and frequent generalized tonic - clonic FS appeared during that time. There were 36 people in 5 generations of the family including 14 patients (8 males and 6 females) ,aged from 4 years and 5 months to 82 years. FS presented in 8 cases (Ⅱ 2, Ⅲ1, Ⅲ4, Ⅲ6, Ⅳ1, Ⅳ11, Ⅳ17, Ⅴ2),febrile seizures plus(FS +) in 4 cases ( Ⅳ2, Ⅳ12, Ⅳ13, Ⅳ14), ES + and absence seizures in 1 case ( Ⅴ1 ), uncertain type in 1 case (Ⅰ2). The results of EEG indicated that 12 cases were normal and 4 cases with FS+ and 1 case with absence seizures had epileptic discharges. Apart form Ⅳ13, Ⅳ14 who were treated with magnesium valproate, the dosage for the other patients decreased, or medicine terminated or without medicine, and all the patients had no recurrence of seizures. The intelligence, movement development and neurological examinations of the family were all normal. Head CT scan of 3 cases were normal. Conclusions GEFS+ is autosomal dominant inheritance disease with conspicuous genetic heterogeneity and phenotypic heterogeneity. The apprehension of GEFS+ plays an important role in diagnosis and differential diagnosis of epilepsy in childhood.