Objective To evaluate the impact of lower renal calyceal anatomic structure on flexible fibreoptic ureteroscopy with holmium laserin treatment of calyceal calculi.Methods From January 2007 to December 2011,a total of 60 patients with a lower calyceal renal stone were enrolled in this study.The mean age was 51 years (range 23 to 78 years).The mean height was 169.8 cm,and mean body mass was 71.2 kg.Intravenous urogram (IVU) was performed on all patients and the lower pole anatomy (including infundibulopelvic angle,length of the inferior caliceal infundibulum and infundibular width) were measured in these patients.The correlation between lower pole anatomy and the success of flexible fibreoptic ureteroscopy with holmium laser for calyceal calculi was analyzed.Results Of the 60 patients,42 patients were successful in stone clearance.The patients in the stone-free group age of (50.1 ± 14.6) years,height (169.8 ±5.1) cm,body mass (71.4 ±5.1) kg,the maximum stone size in diameter (10.9 ±2.1) mm,stone burden (85.4 ± 9.5) mm2,lower infundibular length (36.3 ± 3.7) mm and lower infundibular width (4.9 ±1.4) mm; the other 18 patients age (50.7 ± 11.7) years,height (169.9 ±6.4) cm,body mass (71.6±4.7) kg,the maximum stone size in diameter (11.3 ±2.4) mm,stone burden (82.5 ±8.6)mm2,lower infundibular length (37.2 ± 2.3) mm and lower infundibular width (4.8 ± 1.9) mm.There was no difference between the stone-free group and the residual group in all above parameters (P ＞ 0.05).However,the infundibulopelvic angle in the stone-free group was significantly greater than that in the residual group (63.4 ± 23.2 vs 45.32 ± 17.6,P ＜ 0.05).x2 test showed the stone clearance rate in patients with angle ≥45 was better than that in those with angle ＜45 (84.6％ vs 42.7％,P ＜0.05).If grouped by infundibulopelvic angle,patients with infundibulopelvic angle greater than 90°had stone clearance rate 92.3％ (12/13),those with angle ranged from 30° to 90° had 73.2％ (30/41),and those with infundibulopelvic angle smaller than 30° had 0％ (0/6).Logistic regression analysis showed that the angle was a significant independent predictor of stone clearance (OR =1.12,P ＜ 0.05).Conclusions The infundibulopelvic angle has adverse influences on the performances of flexible ureteroscopy.The samller the angle is,the poorer the performances of flexible ureteroscopy is.

Objective To evaluate the performance of community health institutions of Pudong new district.Methods According to the criteria of the Ministry of Health of China,the performance evaluation system appropriate for local area was developed.A cross-sectional survey was conducted at 44 community health centers,and data was analyzed using descriptive statistics,correlation coefficients and multi-factor linear regression.Results The average score of total performance of these 44 community health centers was 78.53.The average scoring rate for institution management,public health service,basic medical care service,CTM service,and comprehensive satisfaction was 68.49％,89.09％,63.51％,87.80％,76.32％,respectively.Degree of informationization (0.477),regional location (0.331) and participating in medical consortia(-0.309)had significant impact on the total performance.Degree of informationization(0.302)and pilots for family doctors(0.301)had significant impact on the basic medical service performance.Conclusion There is a tremendous room for performance improvement for community health institutions in Pudong.Regional location and degree of informationization were the most crucial factors affecting the performance,irrelevant to institutional sizes.Proposals were raised for strengthening the construction of informationization,expanding pilots for family doctors,perfecting the mechanism of medical consortia.

〔Abstract〕 The paper explains the construction background and objective of the graded diagnosis information system based on the provincial health information platform in Zhejiang province and introduces the system framework and function in detail .This system pro-vides uniform referral information service for medical institutions at all levels all over the province and realizes the exchange and sharing of dual referral records between basic health service institutions and big hospitals .

Objective:To predict and identify HLA-A * 0201 restricted CD8+ CTL epitopes in Mycobacterium tuberculosis (Mtb) antigen Ag85C, so as to provide evidence for epitope-based study for tuberculosis (TB) vaccine. Methods: The online database SYFPEITHI was applied to predict the potential HLA-A * 0201 restricted epitopes from Ag85C, an antigen of Mycobacterium tuberculosis. T2 cell line was used to assay the affinity between the predicted peptides and HLA-A * 0201 molecules. The specific CTL lines were induced from peripheral blood mononuclear cells (PBMCs) of HLA-A * 0201 positive TB patients and PPD+ healthy donors by peptides with high binding affinity to HLA-A * 0201 molecules. IFN-?production, in vitro proliferation and cytotoxicity of peptide-induced CTL were determined to screen HLA-A * 0201 restricted CD8+ CTL epitopes from those candidates. Results: Fourteen potential epitopes were identified from the SYFPEITHI database. After binding affinity assay, 3 of the 14 peptides (170-178 aa, 317-325 aa, and 144-153 aa) were found to have high binding affinity to HLA-A* 0201 molecules. However, only one peptide (144-153 aa) stimulated its specific CTL to release IFN-y, proliferate in vitro and produce specific cytotoxicity. Conclusion: We have successfully identified a HLA-A * 0201 restricted CD8+ CTL epitope of Mtb Ag85C-FLTREMPAWL( 144-153 aa) , which might be a candidate epitope for TB vaccine designing. Our findings provides a basis for developing novel and effective anti-TB vaccine.

AIMTo study the chemical constituents of the stems of Opuntia vulgaris Mill(Cactaceae).

METHODSThe compounds of Opuntia vulgaris were isolated by chromatography of Amberlite Dowex 50 and silica gel, and identified by means of UV, IR, MS, 1D and 2D NMR.

RESULTSThree compounds were isolated and identified as: opuntin B(I), 4-hydroxyproline(II) and tyrosine(III).

CONCLUSIONCompound I is a new alkaloid.

Hydroxyproline ; chemistry ; isolation & purification ; Maleimides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Opuntia ; chemistry ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Tyrosine ; chemistry ; isolation & purification

To explore the molecular mechanisms of acute promyelocytic leukemia cell differentiation induced by cAMP combined with low-dose As2O3, the PR9 cell line, which was stably transfected by PML-RARa fusion gene, was used as in vitro model. The effects of PML-RARa on cAMP-induced AML cell differentiation were evaluated according to cell growth, cell morphology, cell surface antigen as well as luciferase reporter gene assay, in the cells before and after the treatment with cAMP and/or As2O3. The results showed that cAMP alone could slightly increase the expression of CD11b in the PR9 cells expressing the PML-RARa fusion protein, but could not induce these cells to differentiate. The cells presented the terminal differentiation morphology and significantly increased CD11b expression only under the treatment of cAMP combined with As2O3. In addition, PML-RARa had strong inhibitory activity on the transcription of the reporter gene containing cAMP response elements. In conclusions, the PML-RARa fusion protein could dramatically block the signaling pathway of cAMP during the AML cell differentiation.
Arsenicals ; pharmacology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cyclic AMP ; metabolism ; pharmacology ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; Oxides ; pharmacology ; Signal Transduction ; Transfection

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo investigate the molecular mechanisms by which IFN-alpha regulated retinoic acid-induced gene G (RIG-G) expression.

METHODSThe expression of STAT1, p-STAT1 and RIG-G in IFN-alpha-treated NB4 cells was detected by Western blot. The roles of STAT1, STAT2 and IRF-9 in IFN-alpha-induced RIG-G expression were analyzed in STAT1-null U3A cells by cell transfection, reporter gene assay, co-immunoprecipitation and chromatin immunoprecipitaion.

RESULTSIn U3A cells, only when STAT2 and IRF-9 were co-transfected, the luciferase activities of RIG-G promoter-containing reporter gene could be highly increased about 8-fold compared with that in the control group. Moreover, in the absence of IFN-alpha, similar effects were observed in either IRF-9 co-transfected with wild type or mutant form of STAT2, whereas IFN-alpha could increase the transactivation activity of wild type STAT2 and IRF-9 by 6-fold compared with that without IFN-alpha, but had no effect on mutant STAT2. In addition, STAT2 could interact with IRF-9 and bind to the RIG-G promoter.

CONCLUSIONSTAT2 may interact with IRF-9 in a STAT1-independent manner. The complex STAT2/IRF-9 is the key factor mediating the expression of RIG-G gene regulated by IFN-alpha. This is a novel signal transduction cascade for IFN which is different from the classical JAK-STAT pathway.

Cell Line, Tumor ; Fibrosarcoma ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Immunoprecipitation ; Interferon-Stimulated Gene Factor 3, gamma Subunit ; genetics ; metabolism ; Interferon-alpha ; pharmacology ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Phosphorylation ; Plasmids ; STAT1 Transcription Factor ; genetics ; metabolism ; STAT2 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo investigate the molecular mechanisms of the expression regulation of retinoic acidinduced gene G (RIG-G) by interferon alpha (IFNalpha).

METHODSRIG-G promoter region was analyzed by bioinformatics. The functional activities of RIG-G promoter with or without IFNalpha were detected by luciferase reporter assay and electrophoretic mobility shift assay (EMSA).

RESULTSRIG-G promoter region contained two well-conserved IFN-stimulated response elements (ISREs). Both ISRE I and ISRE II showed their effective binding abilities with signal transducer and activator of transcription 1 (STAT1). In HT1080 cells, in contrast with the empty plasmid pXP2, pXP2-A reporter construct containing intact ISRE I and ISRE II showed a significant higher baseline expression (1741.2 +/- 517.5) which could be further enhanced up to three-four folds by IFNalpha (5338.7 +/- 1226.9, P < 0.05). However, the luciferase activity of pXP2-A as well as its IFNalpha inducibility could be abrogated in STAT1-deficient U3A cells (from 1741.2 +/- 517.5 to 406.1 +/- 103.2, P < 0.05), indicating that the STAT1 protein was a prerequisite for the activities of ISRE I and ISRE II.

CONCLUSIONISREs present in RIG-G promoter region are molecular basis of IFNalpha induced RIG-G expression. RIG-G is a target gene directly regulated by STAT1 protein and should play a key role in IFNalpha signaling pathways.

Base Sequence ; Cells, Cultured ; Gene Expression Regulation ; drug effects ; physiology ; Humans ; Interferon Regulatory Factor-1 ; genetics ; metabolism ; Interferon Regulatory Factors ; genetics ; metabolism ; Interferon-alpha ; pharmacology ; physiology ; Interferons ; physiology ; Molecular Sequence Data ; Promoter Regions, Genetic ; drug effects ; genetics ; physiology ; STAT1 Transcription Factor ; metabolism

OBJECTIVE:To explore an effective method to formulate management-related strategies for off-lable use of drugs by the evidence-based medicine. METHODS:The process of guideline formulation included seven procedures,i.g. establishment ofguideliesformulation workgroup;investigation and selection of the status quo on off-label drug use;identification of the clinical problems;retrieval and evaluation and comprehensing of evidence;applification of GRADE in evidence quality grading;formation of the recommendations consensus;peer review and result publication. And eventually guidelines were formed based on the steps. This study took off-label use of rheumatoid immunoprotective subjects as a case to explore. RESULTS & CONCLUSIONS:Based on the evidence evaluation system and above 7 steps,the methods and process of guideline formulation on off-label use of rheuma-toid immunoprotective subjects that integrated administration,law,clinical medicine,pharmacy subjects were made .The process of guideline formulation fully reflects multidisciplinary characteristics of the workgroup,the advanced nature of the process,the comprehensiveness of evidence ,the rigor of evidence quality grading,and the normalization of consensus. It provides reference in methodology for establishing a comprehensive evidence-based evaluation and management system of off-label use of drugs for all clinical specialist disease. Therefore,this scientific research results may promote the standardization and legalization of the off-label use of drugs management in China.