Objective:To investigate the efficiency of carbon nanotube(CNT)-PAMAM mediated entrance of anti-survivin oligonucleotide into HepG2 cells,and its effects on the proliferation of HepG2 cells.Methods:CNT-PAMAM-anti-survivin oligonucleotide compounds were prepared and characterized by AFM and 1% agarose gel electrophoresis analysis.TEM was used to observe the distribution of CNT-PAMAM-ASODN compounds in HepG2 cells.CNT-PAMAM-ASODN compounds were added into the medium and co-cultured with HepG2 cells for 24 h,48 h,72 h,and 96 h at 37℃,5% CO_2.MTT method was used to detect the effects of ASODN and CNT-PAMAM-ASODN on the proliferation of HepG2 cells.Results:CNT-PAMAM-ASODN compounds were successfully synthesized via AFM and agarose gel electrophoresis.TEM showed that the compounds were located in the cytoplasm.When CNT-PAMAM-ASODN(1.0 ?mol/L)and ASODN(1.0 ?mol/L)were used for a 48 h culture,the inhibitory rates of HepG2 cells were(45.97?4.28)% for CNT-PAMAM-ASODN compounds group,(9.33?0.85)% for ASODN group,and(6.37?0.69)% for CNT-PAMAM group.CNT-PAMAM-ASODN compounds at 1.5 ?mol/L inhibited HepG2 cells by(70.22?7.25)%,and the inhibitory effects were in a time-and concentration-dependent manner.There was statistical difference between experiment group and control group(P

Coprinus comatus cultivated on cotton seed hull medium decomposed lignocellulose straggly and was high of absolute biological efficiency. Lignocellulose is the main carbon source for the fruiting stage of the fungus. There existed the positive correlation between the degradation rates of the cellulose and hemicellulose in the medium and the activities of extracellular CMCase (carboxymenthelcel lulase), FPase (filter paper cellulase) and HCase (hemicellu-lase), there also existed the positive correlation between the degradation rate of the lignin in the medium and the activity of extracellular laccase, but no correlation between the degradatio rate of the lignin in the medium and the activity of peroxidase. The activity of extracellular amylase was comparatively high at mycelial growth stage, and the protease activity peek was at teh time when the fruitbody matured.

Objective To evaluate the value of ultrasound (US) in assessment knee joint inflammation in patients with juvenile rheumatoid arthritis(JRA).Methods US scans of the knees obtained in 30 children at clinically active stage; JRA was compared with those obtained in 30 healthy children and 10 JRA patients in clinical remission.Results Changes in synovial membrane thickness and presence of fluid in suprapatellar bursa showed statistically significant differences between JRA patients with active disease and the other subjects.Alterations in contour of the articular cartilage were demonstrated in 10 knees of patients with JRA.Conclusion US is a simple sensitive and reliable methods for the assessment and monitoring of knee joint involvement in JRA.

Objective:To investigate the effect of blocking BRCAA1 gene expression with siRNA on the proliferation of tumor cell line MCF-7 and Rb gene expression.Methods:RNAi was employed to specifically knock down BRCAA1 expression.MCF-7 cells were transfected with complexes constructed with lipids and chemically synthesized Pre-designed anti-BRCAA1 siRNAs.The total RNA was isolated and reversely transcribed after 48 h.The expressions of BRCAA1 and Rb mRNA were determined by Real-Time PCR.Results:Compared with negative control,transfected MCF-7 cells had a 42.3% decrease in expression of BRCAA1 mRNA and an 11.1% increase in Rb mRNA expression.The inhibitory rate of MCF-7 cells proliferation was(81.6?6.1)%.Conclusion:There may be some antagonistic effect between BRCAA1 gene and Rb gene in proliferation of tumor cells,which provides a potential target for anti-tumor gene therapy.

objective To reveal the enterovirus infection within children suffering hand-foot-mouth disease(HFMD)in the Capital Institute of Pediatrics from Aprial to August,2009,for the sake of clinical diagnosis and treatment.Methods Beth throat swab and vesicle fluid were taken respectively from 159 children with HFMD.And RNA were extracted from each sample followed with real-time fluorescence quantitative RT-PCR kits with three reagents:universal enterovirus primer,Coxsackievirus A16(CAl6)primet and enterovirus 71(EV71)primer.Parts of postivive samples were sequenced and analyzed.Results(1)EV genes were detected from 152cases,of which,102 cases were positive for CA16 and 43 were positive for EV71.(2)CV16:EV71 was 2.37:1.The positive rates of throat swabs and vesicle fluid samples were not statistically significant.(3)The PCR results were same with that of sequence analysis.Conclusion The hand-foot-mouth disease recently appeared in our hospital was mainly related to the EV71 or CA16 infection.And the percentage of EV71 infectious obviously increased compared to that of 2007.

Objective To analyze the pathogen and characteristics of the serum types of enterovirus of hand-foot-and-mouth disease ( HFMD ) in the summer,2009.Methods Both throat swab and herpes fluids were taken respectively from 174 children with HFMD in the outpatient infection during April to September,2009.Anti-Cox A16 and anti-EV71 IgMs in the serum were detected with ELISA.And RNA were extracted from each sample followed with real-time fluorescence quantitative RT-PCR kits with three reagents:universal enterovirus primer,Coxsackievirus A16 (CA16) primer and enterovirus 71 (EV71)primer.Parts of positive samples were sequenced and analyzed.Results ( 1 ) EV genes were detected from 167 cases,of which,112 cases were positive for CA16 and 46 were positive for EV71.CA16:EV71 was 2.43∶ 1.(2)There were 51 cases with CA16 IgM positive and 25 cases with EV71 IgM positive in the early collected sera,and in the later samples,98 cases with CA16 IgM positive and 32 cases with EV71 IgM positive.(3)The nucleotide homologies were 88.7％-98.5％ of VP1 gene among CA16.The nucleotide homologies were 94.9％-99.7％ of VP1 gene among EV71,and were 92.1％-95.3％ with C4 subtype.Conclusion The mainly pathogen causing HFMD in children in the summer,2009 were CA16 and EV71.EV71 infection,mainly C4 subtype,was highly elevated according to the earlier reported.Real-time RT-PCR is more appropriate than the serological test.

OBJECTIVETo study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor beta subunit (PDGFR- beta) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs.

METHODSExpression vector of anti-PDGFR- beta ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR- beta, alpha-smooth muscle actin (alpha-SMA), and type I and type III collagen was detected by means of northern blot, Western blot and immunocytochemical staining respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was demonstrated with flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.

RESULTSThe expression of PDGFR- beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSCs only 43% to 51% of that in control cells (t > or = 3.957, P < 0.05), and alpha-SMA expression level, type I and type III collagen synthesis ability were also reduced (t > or = 6.790, P < 0.01). The proliferation of ribozyme-transfected HSCs was significantly decreased (t > or = 3.858, P < 0.05), and the proliferation response to PDGF BB was markedly inhibited. However the apoptotic rate was significantly increased in ribozyme-transfected HSCs (chi2 > or = 14.157, P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy.

CONCLUSIONSThe anti-PDGFR- beta ribozyme can be expressed stably in HSCs, cleave the target RNA effectively, inhibit HSCs proliferation and collagen synthesis, and induce HSC apoptosis. The results suggest that inhibiting PDGFR- beta expression in HSCs may be a new therapy for liver fibrosis.

Apoptosis ; drug effects ; Cell Division ; Cells, Cultured ; Hepatocytes ; drug effects ; physiology ; Humans ; Liver ; pathology ; RNA, Catalytic ; pharmacology ; RNA, Messenger ; biosynthesis ; Receptor, Platelet-Derived Growth Factor beta ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the prevalence of human papillomavirus (HPV) and the HPV genotype distribution in invasive squamous cell carcinoma of the uterine cervix in the Mongolian women in Inner Mongolia autonomy region.

METHODSThe prevalence data of HPV in our department were retrospectively reviewed. INNO-LiPA genotyping technique was used to detect HPV genotypes in the reserved carcinoma tissue specimens.

RESULTSTotally 63 tissue specimens were collected and detected. The prevalence of HPV was 93.7%. The positive rates of HPV among different clinical staging and different pathological grading were not significantly different (P >0.05). The prevalence of HPV16 was not significantly different among different age groups (P>0.05). HPV16 (69.8%), HPV18 (4.8%), HPV31 (4.8%), HPV39 (4.8%), and HPV52 (3.2%) were the 5 dominating HPV genotypes in all cases.

CONCLUSIONSHPV infection is closely correlated with invasive squamous cell carcinoma of the uterine cervix in Mongolia women. HPV16 is the most important genotype in invasive squamous cell carcinoma of the uterine cervix, followed by HPV18, 31, and 39. HPV infection dose not affect the progression and differentiation of invasive squamous cell carcinoma of the uterine cervix.

Adult ; Aged ; Asian Continental Ancestry Group ; Carcinoma, Squamous Cell ; virology ; Female ; Genotype ; Humans ; In Vitro Techniques ; Middle Aged ; Papillomaviridae ; classification ; genetics ; Papillomavirus Infections ; genetics ; virology ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; virology

BACKGROUNDActivation and proliferation of hepatic stellate cells (HSC) is essentially involved in the development and progression of hepatic fibrosis. The most potent growth factor for HSC is platelet-derived growth factor receptor (PDGF) and PDGF receptor beta subunit (PDGFR-beta) is the predominant signal transduction pathway of PDGF which is overexpressed in activated HSC. This study investigated the cleavage activity of hammerhead ribozyme targeting PDGFR-beta mRNA in HSC and the effect on biological characteristics of HSC.

METHODSExpression vector of anti-PDGFR-beta ribozyme was constructed and transfected into rat activated HSC with lipofectamin. The positive cell clones were gained by G418 selection. The expression of PDGFR-beta, alpha-smooth muscle actin, and typeI and type III collagen were detected by using Northern blot, Western blot and immunocytochemical staining, respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was analyzed by using flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.

RESULTSThe expression of PDGFR-beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSC by 49% - 57% (P < 0.05 - 0.01). The proliferation and alpha-smooth muscle actin expression of ribozyme-transfected HSC were significantly decreased (P < 0.05 - 0.01), and the type I and type III collagen synthesis were also reduced (P < 0.01). In addition, the proliferative response of ribozyme-transfected HSC to PDGF BB was significantly inhibited. Otherwise, the apoptotic cells were significantly increased in ribozyme-transfected HSC (P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy.

CONCLUSIONSThe anti-PDGFR-beta ribozyme effectively cleaved the target RNA and significantly inhibited its expression, which blocked the signal transduction of PDGF at receptor level, inhibited HSC proliferation and collagen synthesis, and induced HSC apoptosis. These results suggest that inhibiting PDGFR-beta expression of HSC may be a new target for the therapy of liver fibrogenesis, and ribozyme may be a useful tool for inhibiting PDGFR-beta expression.

Actins ; biosynthesis ; Animals ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Collagen ; biosynthesis ; Liver ; cytology ; Liver Cirrhosis ; drug therapy ; pathology ; RNA, Catalytic ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptor, Platelet-Derived Growth Factor beta ; genetics

OBJECTIVETo investigate the relationship between renal tubular cells transdifferentiation and chronic renal interstitial fibrosis induced by Fangchi Extract in rat.

METHODThe chronic renal interstitial fibrosis rat model was made by giving Radix Aristolochiae Fangchi extract (RAFE) and aristolichic acid (AA) respectively to rats through infusing stomach about 22 weeks discontinuously. Through immunnal histochemistry methods, investigating the expression of symbol proteins: Cytokine( CK) , alpha-Smooth muscle actin ( alpha-SMA) and Vimentin, and also the important fibrosis inducing factor-Transforming growth factor-beta (TGF-beta1 )on renal tubular cells.

RESULTIn RAFE and AA Groups, the expression of CK on renal tubular cells is declined comparing with the Control Group, and the enhanced expression of alpha-SMA and Vimentin can be observed on tubular cells. The expression of TGF-beta1 on renal tubular cells stronglhy increased, too.

CONCLUSIONPart of the renal tubular cells was transdifferentiated into myofibroblasts. Renal tubular cells may participate the occurance of chronic renal interstitial fibrosis, TGF-beta1 may accelerate the transdifferentiation of tubular cells.

Actins ; metabolism ; Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; isolation & purification ; toxicity ; Cell Transdifferentiation ; drug effects ; Cytokines ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Fibrosis ; Kidney Diseases ; chemically induced ; metabolism ; physiopathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Plant Extracts ; isolation & purification ; therapeutic use ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; Vimentin ; metabolism