This article reviewed the gene markers and the sequences for the detection of the species and strains of Mycobacteria tuberculosis. The gene markers and the sequences conclude insertion sequences, tandem repeat sequences, the polymorphic GC-rich repetitive sequences contained in the plasmid pTBN12, 36 bp direct repetitive sequences, spacer oligonucleotides.

Objective To isolate and purify the membrane protein of SeAx cells, which was derived from cutaneous T-cell lymphoma cell line (CTCL), and to prepare polyclonal antibodies against membrane proteins. CTCL antigens were screened with SEREX method from the cDNA expression library of SeAx cells. Methods SeAx cells were cultured to 2?1010 and membrane proteins were purified by means of differential centrifugation after homogenate was made, and organelle protein specific antibodies were used to perform Western blot. Rabbit was immunized 4 times by the membrane proteins, and the serum antibody titer was measured by ELISA after each immunization. The specific interactions between antibodies in serum and SeAx proteins were detected by Western blotting. E. coli were initially transfected by recombinant phages, and the recombinant proteins expressed were transferred onto the nitrocellulose membranes, which were then again reacted with diluted serum from the immunized rabbit. Positive clones were subcloned and the nucleotide sequence of the cDNA inserts was determined. Finally, cDNA sequence analysis via database searching was performed to define the gene. Results Membrane proteins of SeAx cells were purified by differential centrifugation and a 1?10-5 titer of antibody was obtained after 4 times of immunization. 1?106 phage clones were screened, 25 of which were positive. Nucleotide sequencing and database searching showed that 4 cDNA inserts were cell membrane proteins, respectively as integrin alpha 4,ligatin, matrix metalloproteinase 24 and MHC-I molecule HLA-A. Conclusion Membrane proteins of CTCL and SeAx cells, are purified and polyclonal antibodies against SeAx cell membrane proteins are successfully prepared. The 4 genes of membrane proteins are identified by means of SEREX from SeAx cDNA library, expressed by phages. These genic candidates are able to induce systemic humoral immune reactions and might therefore be promising targets for immunotherapeutic interventions of CTCL.

Objective Human immunoglobulin combinatorial library was generated by using phage surface display expression system, and phage antibodies (Fabs) to platelet were screened.Methods:PBMC were separated from a patient who Were transfusion refractory to platelet. mRNA was isolated and cDNA was synthesized by reverse transcriptase.The immunoglobulin heavy chain Fd and the light chain ? genes were amplified by half nested PCR and the PCR products were cloned into the expression vector pCome3 respectively.The immunoglobulin combinatorial library was constructed and screened by 3 rounds of affinity selection.Results Human Immunoglobulin Combinatorial Library was successfully constructed.The specific phage antibodies were highly enriched after 3 rounds of biopanning selection against platelet membrane proteins.Conclusion The antibody library and human monoclonal antibodies to platelet may be useful as molecular tools to study the anti platelet drugs, platelet related diseases, epitope of human platelet antigens.

Objective To investigate the changes in Bcl-2-associated athanogene 1(BAG-1)expression,and the mechanism of nuclear translocation in rat alveolar macrophages(AMs)induced by lipopolysaccharide(LPS)and dexamethasone(Dex).Methods Primary culture AMs treated by LPS and Dex were divided randomly into three groups:6h group,2h group and 24h group.The BAG-1 expression in AMs was detected with Western blot.The interactions between BAG-1 and glucocorticoid receptor(GR)were detected with immune co-precipitation.The changes in GR expression in nuclear protein were evaluated with Western blotting after transfection of RNA interference recombinant plasmids(named psilencer 3.1-GR)targeting to GR gene.Results The expression of BAG-1L in total protein increased,and that of BAG-1S showed no changes.Only BAG-1L,with no BAG-1S,was detected in nuclear protein,and its expression increased gradually in 24h.Interaction between BAG-1L and GR was found in nucleolus after treatment.After transfection of plasmids psilencer 3.1-GR,the BAG-1L expression in nuclear protein decreased significantly compared with that of non-transfection group(P

In recent years,with the completion of the Human Genome Project and the development of mapping the Human Ge?nome Methylation Variable Site Map Plan,research on epigenetics and the generation and deveopment of cancer,epigenetic treatment drugs,especially the successful clinical application of the DNA methyltransferase and histone deacetylation inhibitors in the treatment of cancer patients,epigenetic has becoming a hot spot. This article reviews the recent progress in pharmacological action of epigenetics-based anticancer drugs,it may provide some new ideas to the therapy and fundamental research of cancer.

Objective To investigate the change of the first-phase insulin secretion in impaired glucose regulation and its influencing factors.Methods The investigation was performed in 1024 subjects who were selected by cluster sampling.The body waist circumference (WC) was examined in these subjects.Oral glucose tolerance test was performed,and fasting plasma glucose,fasting insulin as well as 30-,60-,and 120-minute plasma glucose and insulin after glucose intake were also tested.All the 1024 subjects were divided into 5 groups based on the results of oral glucose tolerance test:normal glucose tolerance group,impaired fasting glucose group,impaired glucose tolerance group,impaired fasting glucos/impaired glucose tolerance group,and diabetes mellitus group.First-phase insulin secretion index (the ratio of change in insulin to change in glucose during the first 30 minutes after glucose ingestion was calculated.Results Compared with normal glucose tolerance group (1.96 ± 1.03),the first-phase insulin secretion index significantly decreased in the impaired fasting glucose group (1.79 ±0.91) (P =0.007),the impaired glucose tolerance group (1.81 ± 0.97) (P =0.007),the impaired fasting glucose / impaired glucose tolerance group (1.59 ± 0.85) (P =0.005),and the diabetes mellitus group (1.30 ± 0.60) (P =0.004).Logistic regression analysis showed that WC was the strongest influencing factor of first-phase insulin secretion (β =0.716,P =0.000).Conclusions The firstphase insulin secretion index has already dropped in the stage of impaired glucose regulation.WC can be a useful indicator for evaluating first-phase insulin secretion.

BACKGROUND:The carriers made from biodegradable and biocompatible polymeric materials represent an exciting approach to increase the bioavailability and stability of oral y administered insulin by the chemical reaction or physical encapsulation of insulin.
OBJECTIVE:To mainly review the research progress of the material types, preparation methods, physicochemical characteristics, in vitro release kinetics, and bioavailability of polymeric materials adopted as oral insulin carriers.
METHODS:The first author searched PubMed, Elsevier and CNKI databases for articles (2002-01/2013-02) concerning the polymeric materials and oral insulin carriers with the key words of“polymeric biomaterials, oral insulin, carrier”in English and in Chinese.
RESULTS AND CONCLUSION:Currently, there are mainly two kinds of polymeric biomaterials used for oral insulin delivery systems, that is, natural polymeric biomaterials (such as chitosan and alginates) and synthetic polymeric biomaterials. The most commonly used synthetic polymeric materials for the preparation of these vehicles are polyesters, polyacrylates and their copolymers, which are wel known for their good biodegradability, biocompatibility, and physiological properties. Although researchers have tried to develop promptly oral insulin formulation using various technologies, the reports about clinical application or commercial success have not been seen because of several questions such as polymer material as a carrier, the lower bioavailability of insulin, the quality standards and stability of the formulation. Hence, future studies wil focus on the development of a new type of polymer-based material as carriers by choosing the new materials or modifying physical and chemical characteristics of existing polymers, to avoid gastrointestinal destruction of the insulin and increase bioavailability of insulin in the body, so as to obtain the good control ed release rate and effect.

Objective: To investigate the expression changes of Bcl-2-associated athanogene 1(BAG-1) and its regulatory effect on the glucocorticoid receptor(GR) activity in rat alveolar macrophages in conditions of cell inflammation and glucocorticoid therapy.Methods: The expression changes of BAG-1 were detected by Western blot after lipopolysaccharide(LPS) and Dexamethasone(Dex) treatment of rat alveolar macrophages(AMs),the interaction between BAG-1 and GR determined by immune coprecipitation experiment,and the transcriptional activation of GR measured by relative luciferase activity assay.Results:After LPS and Dex treatment,the expression of BAG-1L in total protein increased but that of BAG-1S remained changed,BAG-1L rather than BAG-1S was detected in nuclear protein and its expression increased gradually within 24 hours,the interaction between BAG-1L and GR was observed in nucleoli,and the transcriptional activation of GR decreased,with a negative correlation between BAG-1L expression and GR activity.Conclusion:LPS and Dex acting on rat alveolar macrophages,the expression of BAG-1L increases,which,coupled with GR,translocates into nucleoli and inhibits GR activity.This might be the important mechanism that underlies glucocorticoid resistance in inflammation.

Objective:To approach the distinctive histopathological chages of nontuberculous mycobacteria lymphadenitis. Methods: The experimental animal model of nontuberculous mycobacteria lymphadenitis was developed and the histopathological changes were observed under the light microscope.Results:The distinctive histopathological changes of nontuberculous mycobacteria lymphadenitis were identified as following: ①Granulomas were found in lymph nodes which were infected with nontuberculous mycobacteria.The coagulation necrosis was located at the center of the granuloma and it was not caseation.Neutrophils were distributed over the necrosis area.Surrounding the center necrosis area,many epithelioid cells,lymphocytes and mononuclear cells could be found.The periphery of the granuloma was surrounded by the fibrous tissues.Outside the granuloma,Langhans giant cells could be found.②Serpiginous necrosis was found in the lymph nodes which were infected with nontuberculous mycobacteria.The shape of serpiginous necrosis was a long strip.In the center area,the strip of coagulation necrosis and the strip of the space which was remnant after the desquamation of necrosis tissues could be found.Many neutrophils were distributed over the necrosis area.Around the center necrosis area,many epithelioid cells,lymphocytes and mononuclear cells could be found.The fibrous tissues were in the borderline.Conclusion:Serpiginous necrosis was one of the distinctive histopathological change of nontuberculous mycobacteria lymphadenitis.

BACKGROUND:With the increasing of aging, the incidence and mortality of osteoporotic hip fracture wil rise. It is of great significance to study the pathogenesis and preventing method. At present, finite element analysis can be used to judge fracture, only for the distribution trend of fracture failure in the starting point or section view, but it cannot completely reflect actual situation of fracture. OBJECTIVE:To build the fracture model of the femoral neck fracture caused by fal ing-induced external force based on the finite element analysis LS-DYNA software, and to evaluate the effect of rupture. METHODS:CT image data of one case of elderly femoral neck fracture were col ected. Using Mimics software, region growth of the contralateral area, cavity fil ing, editing, rebuilding the contralateral proximal femur model were conducted. Data were imported in Hypermesh and LS-DYNA software for meshing, and defining material properties. The failure parameters and interfacial properties were set. The load and force boundary constraints simulating the fal ing were simulated. The model of femoral neck fracture was calculated. Rupture effect was evaluated. RESULTS AND CONCLUSION:(1) The validity of contralateral proximal femur three-dimensional model was verified. Based on the finite element analysis software LS-DYNA, the femoral neck fracture model matched the actual fracture line to a degree of close to 83%. (2) Above results confirmed that based on the finite element analysis, LS-DYNA software can wel simulate the femoral neck fracture, which provides experimental basis to the exploration of femoral neck fracture classification mechanism caused by different fal ing-induced external forces.