In this study, the general bacterial probe and specific cellulolytic bacterial probes were used to quantify the bacteria in rumen. The total RNA were extracted and then hybridized with general bacterial probe after a dilution of concentration. The result showed that there was a high correlation between the hybridization signal and the dilution of total bacterial RNA. Based on the result above, the quantities of three cellulolytic bacteria in rumen sample were detected. The comparative RNA percentage of three cellulolytic bacteria to total bacterial RNA were similar to the previous reports. It can be concluded that the quantification of bacteria in rumen could be conducted by this approach, and which could be used in future research.

The effects of the concentration of sulfuric acid and the ratio of liquid to solid on xylose yield from sugar cane bagasse in its hemicellulose hydrolysis process were studied with the Quadratic Rotary Combination Design. Regression analysis showed that there was a marked regression relationship between the two factors and xylose yield. As the result of optimizing the hydrolysis conditions by regression equation, xylose yield of 24 g/100 g sugar cane bagasse was obtained when sulfuric acid concentration was 2.4 g/L and liquid to solid ratio was 6.2 under the conditions of stream pressure of 2.5 x 10(4) Pa and hydrolysis time of 2.5 h. The macroporous resin adsorption was proved to be a good method to reduce the concentration of yeast cell growth inhibitor in sugar cane bagasse hemicellulose hydrolysate and to enhance the hydrolysate fermentability. The hydrolysate treated with macroporous resin adsorption under pH2 was used as the substrate for xylitol production by a xylitol-producting yeast, Candida tropicalis AS2.1776. At an initial xylose concentration of 200 g/L, all xylose was consumed within 110 h with a xylitol production rate of 1.15 g/L.h, and a xylitol yield of 0.64 g/g xylose.
Candida tropicalis ; metabolism ; Cellulose ; metabolism ; Fermentation ; Hydrogen-Ion Concentration ; Hydrolysis ; Polysaccharides ; metabolism ; Regression Analysis ; Saccharum ; metabolism ; Xylitol ; biosynthesis

OBJECTIVETo investigate the effects of mTOR siRNA on mTOR-p70S6K signaling pathway in esophageal squamous cell carcinoma (ESCC) cells in vitro,and growth and apoptosis in transplanted tumor in nude mice.

METHODSmTOR siRNA was transfected into ESCC cell line EC9706 cells. The expressions of factors of the mTOR/p70S6K signaling pathway were detected by RT-PCR and Western blot. DNA contents and cell apoptosis were determined by flow cytometry, and cell proliferation was measured by CCK-8 assay. The effects of mTOR siRNA on the transplanted tumor growth were assessed in nude mice.

RESULTSThe levels of mTOR and p-p70S6K were significantly decreased (P < 0.05) while the level of p70S6K was increased (P < 0.05) in the cells transfected with mTOR siRNA, compared with that in untransfected cells and cells transfected with control siRNA. After being interfered by mTOR siRNA, the number of apoptotic cells was increased, cell proliferation became slower and cell cycle was arrested in G(1) phase compared with that in control cells. Also, mTOR siRNA inhibited the growth of transplanted tumor in vivo.

CONCLUSIONSmTOR siRNA can effectively interfere in mTOR-p70S6K signaling pathway, induce cell apoptosis and inhibit cell proliferation and tumor growth, suggesting that mTOR-p70S6K signaling pathway plays an important role in the carcinogenesis and development of esophageal squamous cell carcinoma.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; enzymology ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; enzymology ; pathology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; pharmacology ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; genetics ; metabolism ; Transfection ; Tumor Burden

OBJECTIVETo assess the effects of LiCl on prostate cancer growth and to explore the underlying mechanisms.

METHODSEffects of LiCl on cell growth in vitro and in vivo were determined by cell counting and xenografts of prostate cancer cells. Alterations in cell proliferation and the expression of DNA replication-related protein were determined by MTT assay, BrdU incorporation and Western blot.

RESULTSCompared to PBS control group, the number of prostate cancer cells (PC-3) were lower treated with 10 mmol/L LiCl, the number was 1.9×10(5), 4.8×10(5) and the difference was significant (P < 0.05). The inhibition rate of cellular proliferation were 50%, 95% and 98%, respectively, in LiCl group, NaCl and KCl control group, the difference was significant (P < 0.05). The A-Value of BrdU incorporation was 1.5, 1.3 treated with 10 mmol/L, 30 mmol/L LiCl, while the A-value of BrdU incorporation was 4 in PBS control group, the difference was significant (P < 0.05). On the protein level, LiCl downregulates expression of cdc 6, cyclins A and cyclins E, and cdc 25C, and upregulates expression of the CDK inhibitor p21(CIP1). The mean volume and weight of xenograft tumor were 50 mm(3) and 296 mg after LiCl intraperitoneal injection, But PBS control group were 180 mm(3) and 957 mg, the difference was significant (P < 0.05).

CONCLUSIONLiCl disrupts DNA replication and suppresses tumor growth of prostate cancer cells in vitro and in vivo.

Animals ; Antineoplastic Agents ; pharmacology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin A ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; DNA Replication ; drug effects ; Humans ; Lithium Chloride ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Nuclear Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Burden ; drug effects ; cdc25 Phosphatases ; metabolism

OBJECTIVEIn this study, human bronchial epithelial cells were inoculated with positive sputum specimens of HBoV. After four days' infection, cytopathic effects (CPE) were observed by inverted microscopy. These viruses all cause typical cell damages such as rounded and shrivelled, fusion and fallout. These damages got quick following increased future degenerations. The other assay result of CPE within the infected cells were observed by inverted microscopy, have typical "owl's eye" plaque and above 90 percent hemadsorption within the infected cells by erythrocytes for hemadsorption technique. The typical fluorescence lump of nucleus within the infected cells was found by indirect immunofluorescence technique.

CONCLUSIONIsolation and identification of HBoV could be done in the human bronchial epithelial cell, and we found some characterizing CPE in the human bronchial epithelial cell after HBoV infection. The above studies pave a way for studying pathogenicity of human bocavirus.

Bocavirus ; physiology ; Bronchi ; cytology ; Cell Death ; physiology ; Cell Survival ; physiology ; Cells, Cultured ; Epithelial Cells ; cytology ; virology ; Fluorescent Antibody Technique, Indirect ; Host-Pathogen Interactions ; Humans ; Microscopy, Fluorescence

No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.
Adult ; Cholagogues and Choleretics ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; drug therapy ; Male ; Middle Aged ; Taurochenodeoxycholic Acid ; therapeutic use ; Treatment Outcome ; Ursodeoxycholic Acid ; therapeutic use

OBJECTIVETo investigate pave a way for studying pathogenicty of HBoV.

METHODSIsolation and cell culture of HBoV by human bronchial epithelial cell line, which was founded in our laboratory. The morphology of the virus were primarily studied with a transmission electron microscope. In addition, transcript mRNA was detected in human bronchial epithelial cells, which was passaged and infected within HBoV, using the reverse-transcription polymerase chain reaction (RT-PCR). The amplified products nucleotide sequence of HBoV were sequencing and sequence analysis.

RESULTSCytopathic effect (CPE) was observed after the aseptic residue of filtration of 2 case sputum specimens with HBoV, which was inoculated to the human bronchial epithelial cell line. The virus particles were observed in the cytoplasm, which were hexagonal or spherical in shape and 18-26 nm in diameter,bulk was 20 nm. cDNA amplicon obtained 295 bp fragment results of electrophoresis bands as same as NS1 region of the conserved matrix gene of publish sequence of HboV. PCR products nucleotide sequence of HboV were compared with corresponding HboV GeneBank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the parvovirus to the species HBoV.

CONCLUSIONIsolation and identification of HBoV could be done in the human bronchial epithelial cell, and we found some characterizing CPE in the human bronchial epithelial cell after HBoV infection. The above studies pave a way for studying pathogenicty of human bocavirus.

Bronchi ; cytology ; virology ; Cell Culture Techniques ; Cell Line ; Child ; Child, Preschool ; Epithelial Cells ; virology ; Human bocavirus ; classification ; genetics ; growth & development ; isolation & purification ; Humans ; Infant ; Male ; Molecular Sequence Data ; Parvoviridae Infections ; virology ; Phylogeny ; Respiratory Tract Infections ; virology ; Virus Cultivation

WU polyomavirus, which was firstly discovered in 2007, is a new human polyomavirus belonging to Polyomaviridae and containing circular double-stranded genomic DNA. In this study, the 278 clinical sputum specimens from children under 5 years old were collected from Wenzhou Medical College affiliated Wenling First Hospital, Zhejiang Province. Based on identification assay of WU polyomavirus previously reported, a WU polyomavirus was identified from clinical samples successfully, the positive rate was 0.4%. The sequences of PCR products were identical to that of VP2 gene and large T antigen gene derived from WU polyomavirus reported. The above results strongly suggested that the WU polyomavirus isolated was firstly found in Chinese children with acute lower respiratory tract infections. This study provides a firm basis for further research of WU polyomavirus.
Base Sequence ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polyomavirus ; genetics ; isolation & purification ; Sputum ; virology

OBJECTIVETo explore the clinicopathological, immunohistochemical and molecular genetic features of intra-abdomen extra-gastrointestinal stromal tumors (EGISTs) and their differential diagnosis.

METHODSNine cases of EGISTs from the abdominal cavity or retroperitoneum which were previously diagnosed as leiomyoma, leiomyoblastoma, or leiomyosarcoma etc. by a panel of antibodies such as CD117, CD34, alpha-SMA, MSA, desmin, S-100, and PGP9.5 from which five cases were detected for c-kit gene mutation.

RESULTSThe tumors occurred in 5 men and 4 women, the age ranged from 38 to 72 years (mean 61.7 years). Four cases arose from the mesentery, two from omentum, two from retroperitoneum and one located at the hilus of the spleen. The size of tumors ranged from 5 cm to 23 cm (mean 12.9 cm) in diameter and the tumor cell components varied: mainly spindle cells (seven cases), epithelioid cells (one case), mixed cells (one case). Tumors expressed CD117 (8/9), CD34 (5/9), alpha-SMA (3/9), MSA (4/9), desmin (0), S-100 protein (1/9) and PGP9.5 (1/9). Of the five cases examined for heterozygous deletion mutation of 11 exon of the c-kit gene two were found positive. Two borderline cases showed long-term survival of 8 years and 11 years, respectively. In seven malignant cases, two showed adverse outcome, one survived 4 years without recurrence, two were lost in follow up and two new cases were still being in followed.

CONCLUSIONSGIST-type stromal tumors can also occur in the abdomen, most cases were borderline or malignant, tumor coagulative necrosis, mitoses >or= 5 per 50 high-power fields and obvious nuclear atypia indicating malignancy. Differential diagnosis of EGIST including benign or malignant smooth muscle tumors, benign or malignant nerve sheath tumors etc.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Leiomyoma ; pathology ; Leiomyosarcoma ; pathology ; Male ; Middle Aged ; Peritoneal Neoplasms ; chemistry ; genetics ; pathology ; Proto-Oncogene Proteins c-kit ; genetics ; Retroperitoneal Neoplasms ; chemistry ; genetics ; pathology

OBJECTIVETo explore the role of interferon (IFN)-alpha/beta receptor beta subunit (IFNAR2) in the patients' response to IFN-alpha therapy as influenced by the grade of chronic hepatic inflammation, and understand the relation of IFNAR2 expression in the peripheral blood mononuclear cells (PBMCs) with HBV infection.

METHODSLiver tissue specimens were obtained from 21 patients with chronic hepatitis B for examination of the hepatic inflammation, and PBMCs were isolated from another 16 patients with chronic hepatitis B and 15 health control subjects. Both the hepatic tissues and PBMCs were examined for IFNAR2 expression using immunohistochemistry.

RESULTSThe 21 patients with chronic hepatitis B were divided into 3 groups according to the severity of hepatic inflammation, namely G(1) (n=3), G(2) (n=7) and G(3) (n=11) groups. The patients in G(3) group showed had significantly higher IFNAR2 expressions in liver (25.1307-/+7.0700) than those of the G(1) (5.6913-/+1.8422) and G(2) (7.4706-/+5.3572) groups (P=0.000). The IFNAR2 levels in the PBMCs, however, did not show significant difference between patients with chronic hepatitis B and the healthy control subjects.

CONCLUSIONIn patients with chronic hepatitis B, IFNAR2 expression level is positively correlated to the severity of hepatic inflammation, and increased IFNAR2 expression in severe hepatic inflammation is therefore likely to result in increased response rate to INF-alpha therapy. The expression of IFNAR2 in the PBMCs is not associated with HBV infection.

Female ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; Liver ; metabolism ; pathology ; Male ; Receptor, Interferon alpha-beta ; blood ; metabolism