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.

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.

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 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

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 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 evaluate the efficacy and safety of micafungin in the treatment of invasive fungal infections (IFI) in patients with acute leukemia.Methods A total of 133 IFI patients with acute leukemia received micafungin 150 mg once daily for 14 days.The clinical and mycological efficacies were evaluated on (7±2) days and(14±2) days of treatment.Meanwhile,the adverse events were recorded.The normally distributed data was compared using analysis of variance and nonnormal distributed data was analyzed using Wilcoxon rank-sum test.Results Among 133 IFI patients with acute leukemia,116 finished the 14-day micafungin treatment.The total clinical efficacy was 94.8% and the total mycological efficacy was 75.0% at (14±2) days of treatment.The fungus eliminate rates were 82.9%,66.7% and 55.6% against Monilia,Aspergillus and others,respectively.The clinical and mycological efficacies of (14±2)-day treatment were both higher than those of (7±2)-day treatment(X2=6.060,34.416.both P<0.05).The clinical efficacy was not related with age,sex,IFI diagnose,types of leukemia and combinative drugs (X2=26.541,P<0.05).The incidence of drug-related adverse events of micafungin was 3%among 133 patients,which included skin rash in 3 eases, diarrhea in 1 case. Only one case was discontinued because of severe skin rash and micafungin was well tolerant in other patients. Conclusion Treatment of micafungin 150 mg daily for 14 days is effective and safe in IFI patients with acute leukemia.

AIM:To investigate the activation of nuclear factor ?B(NF-?B) induced by lipopolysaccharides(LPS) in rat alveolar macrophages(AMs) and its regulatory role in tumor necrosis factor(TNF-?) secretion.METHODS:The dynamic activity changes of NF-?B induced by LPS were determined with electrophoretic mobility shift assay(EMSA).Antisense oligonucleotides of NF-?B subunit(p65) were transfected into AMs prior to LPS stimulation.The effect of antisense oligonucleotide transfection on expressions of p65 and TNF-? in supernatant were measured with Western blotting and enzyme linked immunosorbent assay(ELISA),respectively.RESULTS:NF-?B activity increased markedly and reached its peak level at 4 h after LPS stimulation.After transfected with antisense oligonucleotides of NF-?B subunit(p65),expression of p65 and TNF-? in supernatant decreased markedly.CONCLUSION:NF-?B activity has a positive effect on regulating secretion of TNF-? in AMs induced by LPS.

Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.