Objective To observe the treatment of ~(99)Te-MDP on typeⅡcollagen induced arthritis (CIA)in rat,and the effect on the expression of synovial MMP-3 and TIMP-1 mRNA.To explore the mech anisms of the ~(99)Te-MDP in the treatment of rheumatoid arthritis.Methods The rats in which C1A(n=24)were divided into three group:the control group(n=8),~(99)Tc-MDP group(n=8)and Methotrexate group(n=8). Arthritis were evaluated by arthritis index and histopathological index and the expressions of MMP-3 and TIMP-1 mRNA in synovium were detected by RT-PCR.Results①The arthritis indexs of the control group, the methotrexate group,the ~(99)Tc-MDP group were increased with time.②The histopathological scnres of the control group were significantly higher than those of methotrexate group and ~(99)Tc-MDP group(P＜0.01).The histopathological scores of cartilage destruction and bone erosion of ~(99)Tc-MDP group were lower than those of methotrexate group(P＜0.05).③The levels of MMP-3 mRNA of the control group,~(99)Tc-MDP group, methotrexate group were notably higher than those of the control group(P＜0.01).The levels of control group was notably higher than that of the ~(99)Tc-MDP group(P＜0.05).There was not significant difference in all groups on the levels of TIMP-1 mRNA(P＞0.05).Conclusions ~(99)Tc-MDP can notably relieve the arthritis symdrome and retard the catilage damage and bone erosion of CIA in rats,and could significantly decrease the MMP-3 mRNA in the synovium.Which may be one of the therapeutic mechanism.~(99)Tc-MDP is better than methotrexate in retarding catilage and bone erosion and decreasing MMP-3 mRNA in CIA rats in a 3-week therapeutic intervention.

of glucose and insulin are also marked after congee intake.

OBJECTIVETo explore the effect of hepatitis B virus (HBV) X protein (HBX) on expression of the host gene Wnt induced secreted protein-1 (WISP-1) that is related to the pathogenic process of hepatocellular carcinoma (HCC).

METHODSTumor and paratumor tissues were collected from HCC patients, and normal liver tissues were collected from healthy controls. Immunohistochemistry was used to evaluate the in vivo presence and expression levels of HBX and WISP-1 in the three tissue types. HepG2 cells stably transfected with pc-DNA3.1(+)-HBX or with pc-DNA3.1(+) only (G0, control) were generated and used to examine in vitro the HBX-induced changes in WISP-1 expression at the mRNA and protein levels by reverse transcription polymerase chain reaction and western blotting, respectively.

RESULTSThe HCC tissues showed significantly higher rates of positivity for WISP-1 expression than the non-tumor controls (76.6% vs. paratumor: 23.4% or normal tissues: 0%, x2= 35.967, P less than 0.01). HBX increased WISP-1 expression in HepG2 cells at both the mRNA (1170.33 +/- 41.26 vs. G0: 265.34 +/- 27.47, t = 31.63, P less than 0.01) and protein (240.33 +/- 11.37 vs. G0: 40.33 +/- 7.09, F = 600.57, P less than 0.01) levels.

CONCLUSIONHBV may up-regulate expression of the host gene WISP-1 through its X protein and thus promote the development of HCC.

Carcinoma, Hepatocellular ; Hepatitis B ; Hepatitis B virus ; metabolism ; Humans ; Liver Neoplasms ; Wnt1 Protein

OBJECTIVETo explore the mechanism by which HBV X gene(HBx) inhibits apoptosis of human hepatoma cell line HepG2 in terms of miRNA.

METHODSThree cell lines were prepared: HepG2 cells stably transfected with HBx (HepG2/HBx), HepG2 cells stably transfected with pcDNA3.1 (HepG2/pcDNA3.1) and HepG2 cells. Flow cytometry was adopted to measure the apoptosis of these three cells and Taqman fluorescence quantitative PCR was used to examine miR-192 expression. After HepG2 cells was transfected with miR-192, the apoptosis was analyzed by flow cytometry and the expressions of p53 and PUMA at mRNA and protein levels were evaluated by SYBR Green quantitative PCR and Western blot, respectively.

RESULTSCompared with HepG2/pcDNA3.1 cells (11.46% ± 0.69%) and HepG2 cells (12.5% ± 0.66%), the apoptosis rate of HepG2/HBx cells (2.37% ± 0.35%) was significantly reduced (F = 171.722, P < 0.01). The level of miR-192 was 49.1% ± 5.9% in HepG2 cells, which was dramatically down-regulated (F = 14.319, P = 0.019) as compared to the other two groups (HepG2/pcDNA3.1: 98.0% ± 8.9%; HepG2: 100%). Compared with HepG2 cells transfected with miR-NC (10.74% ± 1.15%), transfection of miR-192 into HepG2 cells led to increased apoptosis (15.74% ± 1.17%) (F = 18.415, P = 0.013) and higher p53 and PUMA expressions at mRNA (p53: 1.68 ± 0.12 vs 0.90 ± 0.09, F = 43.115, P = 0.003, PUMA: 1.66 ± 0.10 vs 0.98 ± 0.06, F = 22.541, P = 0.009) and protein (p53: 3.07 vs 1, PUMA: 2.13 vs 1) levels.

CONCLUSIONHBx could inhibit apoptosis of HepG2 cells through down-regulation of miR-192 which induces apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Down-Regulation ; Genes, Viral ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; metabolism ; Trans-Activators ; genetics ; metabolism

Objective To evaluate the influences of susceptibility interpretation of Escherichia coli,Klebsiella pneumonia and Proteus mirabilis in China mainland according to the old and new ceftazidime,cefotaxime and ceftriaxone breakpoints in CLSI M100-S20 and CLSI M100-S19. Methods First, We analyzed the antibacterial susceptibility results of the three bacteria by agar dilution method in the SEANIR surveillance item, which were collected from 15 national hospitals between the year of 2005 and 2007 and excluded the AmpC enzyme positive isolates according to the PGR-DNA sequencing method and/or the antibacterial susceptibility phenotype. ESBL phenotype was confirmed by the CLSI phenotypic confirmatory test. Antibacterial susceptibility of the total 2733 Escherichia coli, Klebsiella pneumonia, Proteus mirabilis isolates was retrospectively analyzed by WHONET 5. 4 software according to the breakpoints of the CLSI M100-S19 (S19) and CLSI M100-S20 (S20). Second, 207 isolates of Peking Union Medical College Hospital with the results of both agar dilution method and disk diffusion method were performed by recurrent analysis. Then we observed the inter-method agreement through the scatter diagram according to the breakpoints of S19 and S20. Results First, as to the ESBL positive Escherichia coli, Klebsiella pneumonia and Proteus mirabili.s, the resistant rate of cefotaxime increased from 65.2% , 55.5%, 14. 6% under S19 (64 μg/ml) to 99. 7%, 96. 2% , 93. 8% under S20 (4 μg/ml). The susceptibility rates decreased from 6. 0%, 11.5%, 33.3% under S19 (8 μg/ml) to 0%, 0. 2%, 0% under S20 ( 1 μg/ml). Ceftriaxone had the same trend as cefotaxime. Though ceftazidime was more active than cefotaxime and ceftriaxone, as to the ESBL positive Escherichia coli and Klebsiella pneumonia, the resistant rates slightly increased from 30. 3%,43. 2% under S19 (32 μg/ml) to42.0%, 56. 0% under S20 (16 μg/ml). The susceptibility rates slightly decreased from 58. 1%, 44. 1% under S19 (8 μg/ml) to 44. 7%, 28.0% under S20 (4 μg/ml). Second,as to the ESBL negative Escherichia coli, Klebsiella pneumonia and Proteus mirabilis, all the susceptibility rates of ceftazidime, cefotaxime and ceftriaxone were between 99. 2%-100. 0%, the resistant rate were between 0%-0. 4%. Third, the S20 MIC breakpoints had a good correspondence with the ESBL phenotype.Fourth, according to the recurrent analysis of MIC testing and disk dilution method, r value was 0. 67,0. 79, 0. 77 for ceftazidime, cefotaxime and ceftriaxone, respectively, and all P value were under 0. 01. The intermethod rates of S19 and S20 were both acceptable. Conclusions If the cefotaxime and ceftriaxone S20 new breakpoints were used, the concordance of antibacterial susceptibility results and ESBL phenotype would increase greatly. The clinician could select proper antibiotics according to the antibacterial susceptibility results and clinical symptoms. It is no longer necessary to edit results for cephalosporins, aztreonam, or penicillins from susceptible to resistant. However, until laboratories implement the new interpretive criteria,ESBL testing should be performed as described in Supplemental Table 2A-S1. The relationship between the new breakpoints of ceftazidime and clinical outcomes need to be further evaluated.

OBJECTIVETo investigate the clinicopathologic features of calcium pyrophosphate dihydrate crystal deposition disease (CPPD-CDD).

METHODSThe clinical and pathologic profiles were retrospectively analysed in 20 cases of CPPD-CDD.

RESULTSCPPD-CDD was far more common in women, most frequently involving joints, especially the knees and presenting with various arthrisis. Abnormally calcified and the articular damages were characteristic features by imageing. Histologically, multifocal indigo granular calcinosis was seen in synovium and sometimes appeared as needle-shaped or rhomboid crystals, which characterized the CPPD.

CONCLUSIONSThough clinical symptoms of CPPD are quite variable, the definite diagnosis can be made by the abnormal calcification and joint damage radiographically and the indigo CPPD crystals histopathologically.

Adult ; Aged ; Aged, 80 and over ; Chondrocalcinosis ; diagnostic imaging ; pathology ; surgery ; Female ; Follow-Up Studies ; Hip Joint ; diagnostic imaging ; pathology ; surgery ; Humans ; Intervertebral Disc ; diagnostic imaging ; pathology ; surgery ; Knee Joint ; diagnostic imaging ; pathology ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Spinal Diseases ; diagnostic imaging ; pathology ; surgery ; Synovial Membrane ; pathology ; Tomography, X-Ray Computed

To investigate the effect of hepatitis B virus X protein(HBx) on CtBP-interacting protein(CtIP) which is an important repair factor of DNA double strand break damage in HepG2 cells induced by bleomycin. A HBx stably expressing HepG2 cell line and a control HepG2 cell line with empty vector transfected were established. After the double strand break (DSB) damage occurred, the mRNA and protein levels of CtIP were detected by Real-time PCR and Western blot assay respectively, cell cycle profiles and apoptotic cell death were determined by a flow cytometry, and the position of CtIP in cells was observed by confocal laser scanning microscopy. It showed that HepG2 cells transfected with hepatitis B virus X gene could stably express HBx protein. After being induced by bleomycin, the percentage of apoptotic cell was 16.90%+/-0.89% in HBx stably expressing HepG2 cell line and 15.30%+/-0.86% in control cell line, respectively (q = 2.074, P is more than to 0.05). While the percentage of death cell was 8.71%+/-0.74% in HBx stably expressing HepG2 cell line and 4.90%+/-0.46% in control cell line, respectively (q = 7.126, P is less than to 0.01). The two cell lines manifested the increase of G2/M arrest and significant difference existed between the two cell lines. HBx down regulated the expression levels of CtIP and its mRNA. The CtIP level was 0.66+/-0.04 in HepG2-HBx cell and 0.73+/-0.05 in HepG2-vec cell, respectively (t = 2.314, P is less than to 0.05). The relative mRNA level was 1.00+/-0.06 in HepG2-HBx cell and 1.23+/-0.08 in HepG2-vec cell, respectively (t = 2. 732, P is less than to 0.05). We also found that CtIP was concentrated in the cell nucleus. The research suggests that HBx may affect DNA-repair pathways by disrupting the expression of CtIP.
Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; Real-Time Polymerase Chain Reaction

OBJECTIVETo obtain the knowledge status on recommended heart failure (HF) guidelines among Chinese physicians.

METHODSQuestionnaire on heart failure including 20 multiple choice questions and 10 fill in the blank questions was designed based on the Chinese guidelines for the diagnosis and treatment of chronic heart failure in 2007 and the Chinese guidelines for the diagnosis and treatment of acute heart failure in 2010. The rate of correct answer for each item was calculated and compared among physicians specialized for cardiovascular diseases and not.

RESULTSThe Questionnaire was completed in 400 physicians, including 208 physician specialized for cardiovascular disease and 192 physicians not specialized in cardiovascular disease. The rate of correct answer for 20 multiple choice questions was lower than 60% in 8 questions, 60%-80% in 8 questions, higher than 80% in 4 questions. The rate of correct answers for 10 fill in the blank questions focusing on the aimed dosage of 10 ACEI/ARB/β-blockers was 49%. The 8 multiple choice questions with correct answer rate <60% are detailed items of myocardial remodeling, symptoms suggestive of HF, diagnosis tools for patients with suspected HF, the AHA stages of heart failure, the Forrester's hemodynamic classes of acute heart failure, the goals of ACEI/ARBs treatment in patients with HF, names of heart diseases which might benefit from ACEI/ARBs treatment defined by evidenced based medicine, and detailed application methods of ACEI/ARBs and β-blockers for HF patients. In general, the rate of correct answer was significantly higher in physicians specialized for cardiovascular disease compared physicians not specialized for cardiovascular disease.

CONCLUSIONThere is a considerable knowledge gap on the Chinese guidelines for the diagnosis and treatment of chronic heart failure and the Chinese guidelines for the diagnosis and treatment of acute heart failure among Chinese physicians. Efforts must be made to educate physicians to improve their knowledge and improve HF patient care.

Adult ; Clinical Competence ; Female ; Heart Failure ; Humans ; Male ; Middle Aged ; Physicians ; Practice Guidelines as Topic ; Surveys and Questionnaires

OBJECTIVEIn order to search the preparation process and optimazing dosage ratio of adsorbed diphtheria-tetanus-acellular pertussis and sabin inactivated poliovirus combined vaccine (DTaP-sIPV), the neutralizing antibody titers of IPV induced by different concentration of DTaP-sIPV were investigated on rats.

METHODSTwo batches of DTaP-sLPV were produced using different concentration of sIPV and the quality control was carried. Together with sabin-IPV and DTaP-wIPV ( boostrix-polio, GSK, Belgium) as control group, the DTaP-sIPV were administrated on three-dose schedule at 0, 1, 2 month on rats. Serum sample were collected 30 days after each dose and neutralizing antibody titers against three types poliovirus were determined using micro-neutralization test.

RESULTSTwo batches of prepared DTaP-sIPV and control sLPV were according to the requirement of Chinese Pharmacopoeia (Volume III, 2005 edition) and showed good stability. The seropositivity rates were 100% for sabin inactivated poliovirus antigen in all groups. The GMTs (Geometric mean titers) of neutralizing antibodies against three types poliovirus increased.

CONCLUSIONThe prepared DTaP-sIPV was safe, stable and effective and could induced high level neutralizing antibody against poliovirus on rats.

Animals ; Antibodies, Viral ; immunology ; Diphtheria-Tetanus-acellular Pertussis Vaccines ; immunology ; Female ; Male ; Poliovirus Vaccine, Inactivated ; immunology ; Rats ; Rats, Wistar ; Vaccines, Combined ; immunology

S-phase kinase-associated protein 2 (Skp2), which plays a role in cell cycle regulation, is commonly overexpressed in a variety of human cancers and associated with poor prognosis. However, its role in nasopharyngeal carcinoma (NPC) is not well understood. In this study, we examined the clinical significance of Skp2, with a particular emphasis on overall survival (OS) and disease-free survival (DFS), in NPC cases in South China, where NPC is an epidemic. Additionally, we explored the function of Skp2 in maintaining a cancer stem cell-like phenotype in NPC cell lines. Skp2 expression was assessed for 127 NPC patients using tissue microarrays and immunohistochemistry and analyzed together with clinicopathologic features, OS, and DFS. Skp2 expression was detectable, or positive, in 75.6% of patients. Although there was no correlation between Skp2 and any clinicopathologic factor, Skp2 expression significantly portended inferior OS (P = 0.013) and DFS (P = 0.012). In the multivariate model, Skp2 expression remained significantly predictive of poor OS [P = 0.009, risk ratio (RR) = 4.06] and DFS (P = 0.008, RR = 3.56), and this was also true for clinical stage (P = 0.012 and RR=3.201 for OS; P = 0.002 and RR=1.94 for DFS) and sex (P = 0.016 and RR=0.31 for OS; P = 0.006 and RR = 0.27 for DFS). After Skp2 knockdown, a colony formation assay was used to evaluate the self-renewal property of stem-like cells in the NPC cell lines CNE-1 and CNE-2. The colony formation efficiency in CNE-1 and CNE-2 cells was decreased. In Skp2-transfected CNE-1 and CNE-2 cells, side population (SP) proportion was increased as detected by flow cytometry. Skp2 is an independent prognostic marker for OS and DFS in NPC. Skp2 may play a role in maintaining the cancer stem cell-like phenotype of NPC cell lines.
Adolescent ; Adult ; Aged ; Carcinoma ; Cell Line, Tumor ; China ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gene Knockdown Techniques ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Staging ; Neoplastic Stem Cells ; pathology ; RNA, Small Interfering ; genetics ; S-Phase Kinase-Associated Proteins ; genetics ; metabolism ; Sex Factors ; Survival Rate ; Tissue Array Analysis ; Transfection ; Young Adult