A new wood-degrading fungus Monodictys asperospera(Cooke & Massee) Ellis with a high level of laccase production was chosen to study.This laccase was purified by ammonium sulfate precipitation,DEAE-cellulose and sephacryl S-300.Purification of about 8.1 fold was achieved with an overall yield of 5.7%.Its molecular weight was estimated to be about 77 kD.The optimum temperature and pH of the lac-case activity were 55?C and 6.0,respectively.Kinetic studies of the laccase showed that the Km and the Vmax for using syringaldazine as substrate was 0.163 mmol/L and 0.194 mmol/(L.min),respectively.The carbo-hydrate content was 18.14%.In addition,it was found that laccase activity was significantly inhibited by Cu2+.

This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.
Animals ; Capsid Proteins ; genetics ; Child, Preschool ; China ; epidemiology ; Evolution, Molecular ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; veterinary ; virology ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Nonstructural Proteins ; genetics

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.
China ; Feces ; virology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

OBJECTIVEto explore the effect of catheter based renal synthetic denervation on renin-angiotensin-aldosterone system (RAAS) and blood pressure reduction in patients with resistant hypertension. and assess the validity and security of the treatment.

METHODSTen patients with resistant hypertension from June 2011 to December 2011 were retrospectively reviewed, and then all of 10 patients screened for eligibility were allocated to renal denervation. Primary endpoints were changes of office blood pressure at 1 week, 1, 3 and 6 months after procedure. We assessed the effectiveness of renal sympathetic denervation with heart rate (HR), renin activity (PRA), angiotensin II (AngII), aldosterone (Ald), and creatinine (Cr) before and 2 weeks after procedure.

RESULTSOffice blood pressure after catheter-based renal denervation decreased by 22.8/9.1 mm Hg (1 mm Hg = 0.133 kPa), 34.8/14.7 mm Hg, 42.6/20.7 mm Hg, 43.2/21.6 mm Hg, at 1 week, 1, 3 and 6 months, respectively (P < 0.001). Meanwhile, the level of PRA, AngII, Ald decreased by (1.11 ± 0.89) ng×ml(-1)×h(-1) (P = 0.003), (17.06 ± 13.82) ng/L (P = 0.004), (404.5 ± 285.8) ng/L (P = 0.002), respectively; and heart rate decreased by 5.1 bpm (P = 0.002). However, the Cr level and eGFR did not change significantly (P > 0.05).

CONCLUSIONCatheter-based renal sympathetic denervation can reduce the level of renin activity, angiotensin II and aldosterone, and causes substantial and sustained blood-pressure reduction.

Adult ; Aged ; Catheter Ablation ; methods ; Female ; Humans ; Hypertension ; metabolism ; physiopathology ; surgery ; Kidney ; innervation ; metabolism ; Male ; Middle Aged ; Renin-Angiotensin System ; Sympathectomy ; methods

OBJECTIVETo screen out main molecular target promoting human neural stem cells (NSCs) of ginsenoside Rg1 by using the gene chip technology.

METHODFirst, MTT assay was adopted to screen out the optimal concentration of Rg1-promoted NSC proliferation (120 mg x L(-1)). Then, on the 7th day after the Rg1-promoted NSC proliferation, the expression of target genes was observed by the gene chip technology. The most important target gene and signal transduction pathways were screened out through the data calculations.

RESULTOn the 7th day after the Rg1-promoted NSC proliferation, obtained 440 differential genes, 266 significantly upregulated genes and 174 significantly down-regulated genes. HES1 gene, CAMP (cyclic adenosine monophosphate)-PKA (protein kinase A) and PI3K (phosphatidylinositol 3 kinase)-AKT signal transduction pathways were closely related to the NSC proliferation.

CONCLUSIONThe differentially expressed genes screened out by gene chip may provide new clues for studies on molecular mechanism of ginsenoside Rg1-promoted NSCs proliferation.

Cell Proliferation ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Neural Stem Cells ; cytology ; drug effects ; metabolism ; Oligonucleotide Array Sequence Analysis ; RNA ; genetics ; isolation & purification

OBJECTIVETo observe the cytotoxic effect of cisplatin with proteasome inhibitor on osteosarcoma cells.

METHODSCell survival was tested by MTT, apoptotic morphology was observed by electron microscopy, apoptotic rate was analyzed by flow cytometry, the transcription level of excision repair cross complementation-1 (ERCC-1) was tested by reverse transcription polymerase reaction.

RESULTSCompared with cells treated with cisplatin alone, cells treated with cisplatin and proteasome inhibitor showed a decreased survival rate, more typical apoptotic morphology, higher apoptotic rate [(14.37 +/-2.37)% vs. (50.93 +/-4.84)%, P<0.01)], and lower transcription level of excision repair cross complementation-1.

CONCLUSIONProteasome inhibitor could increase the cytotoxic effect of cisplatin on osteosarcoma cells and promote cisplatin-induced osteosarcoma apoptosis. These effects may be associated with the decreased transcription of excision repair cross complementation-1.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bone Neoplasms ; pathology ; Cisplatin ; pharmacology ; Drug Synergism ; Oligopeptides ; pharmacology ; Osteosarcoma ; pathology ; Rats ; Tumor Cells, Cultured

Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Humans ; Infant ; Rotavirus ; Rotavirus Infections ; epidemiology

Objective To evaluate the role of late course accelerated fractions.ted irradiation(LCAF) combined with concurrent chemotherapy in the management of esophageal carcinoma.Methods From March 1998 to July 2000,111 eligible patients were randomized into LCAF alone group(LCAF,57 patients)or LCAF plus concurrent chemotherapy group(LACF-CT,54 patients).The radiotherapy regimen was identical in the two groups,consisting of conventional fractionation in the first 2/3 course and accelerated fractionation in the second 1/ 3 course to a total dose of 68.4 Gy/41 Fx/44 d.Chemotherapy regimen consisted of four eycles of cisplatin 25 mg/ (m~2?d)plus fluorouracil 600 mg/(m~2?d)on day 1 to 3 every 4 weeks and was delivered on the first day of radiotherapy.Results The median follow-up time was 67.1 months(range 47.6-76.4 months).The 1-,3-,5- year survival rate was 67%,44% ,40% and 77%,39% 28% in LACF-CF and LEAF group,respectively(P =0.310).Grade 3+4 acute side-effact was 42% and 25% in LCAF-CT and LCAF group,respectively(P＜0. 05),with 3 treatment-related deaths in the LCAF-CT group.Conclusions Late course accelerated fractionated irradiation combined with concurrent chemotherapy has a trend towards improving the survival,at the cost of increasing acute side-effect.Its role needs further confirmation by larger sample studied in randomization.

Objective To detect the ratio of CD19+-CD25+ and CD19+-CD25-B lymphocytes and content of IgA,IgG,IgM and complement C3 in patients with acute cerebral infarction and study their clinical significance. Methods Disease were diagnosed according to the history and cranlal computer tomography or magnetic resonance imagine.Venous blood of 69 cases with acute cerebral infarction and 115 cases with cerebral hemorrhage, 41 cases in normal control group was extrdcted. The ratio of CDl9+-CD25+and CD19+-CD25-B lymphocytes was determined by flow cytometry and content of IgA,IgG,IgM and C3 was measured with scattering turbidimetry.Changes in humoral immunological function were compared among patients with different courses of disease, imaging scores and neurological function scores. Results Differences in CD19+-CD25+and CD19+-CD25-B lymphocytes, IgA, IgG,IgM and C3 were not significant at the acute stage between cerebral infarction and cerebral hemorrhage (P＞0.05,for all).The ratio of CD19+-CD25+B lymphocytes and content of IgG and C3 at the acute stage of cerebral infarction were all higher than that at the recovery stage and in the control group (P＜0.05, for all). There was no statistical signmcance in humoral immunological indices between that at the recovery stage of cerebral infarction and in the control group (P＞0.05, for all). The ratio of CD19+-CD25+ and CD19+-CD25-B lymphocytes was significantly different among patients with different imaging scores (P＜0.05,for all).Neurological function scores at the acute stage of cerebrdl inflarction were not correlated with humoral immunological indices(P＞0.05,for all). Conclusions Same changes occur to humoral immunological function in patients with cerebral infarction and cerebral hemomlage, which might be related with stress,and location and scope of lesions.The larger the lesion of cerebral infarction is,the more obvious changes of humoral immunological function become; with the disappearing of stress,humoral immunological function gradually recovers.