A comparative study on the glycolic acid oxidase (GO) of the domestic Spirulina(Arthrospira) platensis (S_(1)) from alkaline lake in Erdos Plateau and the imported S. (A.) platensis (S_(2)) and S. (A.) maxima (S_(3)) as well is made with colorimetric method. The results show that activity of GO (25℃, pH 8.0) of S_(1), S_(2 )and S_(3) is 70.9 U/gFW, 59.6 U/gFW and 80.9 U/gFW respectively; the GO's optimum temperature of S_(1)、S_(2) and S_(3 )is 30℃; the GO's optimum pH value of S_(1 )is 8.6,while that of S_(2 ) 8.2 and that of S_(3) 8.4; the GO of S_(1 )is stable from 0℃ to 35℃ and from pH 7.6 to pH 10.0, while that of S_(2) from 0℃ to 30℃ and from pH 8.0 to pH 9.0 and that of S_(3) from 0℃ to 35℃ and from pH 8.0 to pH 8.6. Adaptive range of S_(1) GO for temperature and pH is wider, and activity at low and high temperature and under strong acidand alkali conditions is higher than that of the imported species.

Objective To study the expression level and clinical significance of bcl-XL gene in childhood medulloblastoma.Methods The expression of Bcl-XL protein and bcl-XL mRNA were determined by immunohistochemical staining and in situ hybridization in 41 samples of medulloblastoma tissues,as well as 20 normal brain tissues.Results The positive rate of Bcl-XL protein(90.2%) and bclXL mRNA(95.1%) in medulloblastoma group were significantly higher than those in normal human brain tissues(all P

OBJECTIVETo explore the complexity of mRNA in the ejaculated sperm from healthy fertile men.

METHODSSemen samples were collected from 10 healthy fathers. The swim-up method was adopted to purify the sperm from possible contamination of somatic cells and the spermatozoal total RNA extracted by Trizol was used for SAGE library analysis.

RESULTSA totle of 21 052 SAGE raw tags were sequenced from 877 clones and 2 712 unique tags that occurred at least twice in the library were given further analysis. 19.7% of the unique tags had no match in the existing SAGE map, representing novel genes. Molecular function analysis revealed 67% of unique tags related to protein binding or nucleic acid binding categories, 41% to catalytic activity, 13% to message transducer activity, and 10% to transporter, structural and transcription regulator activity, respectively.

CONCLUSIONThere exists a complex repertoire of mRNAs in the ejaculated spermatozoa from fertile men.

Adult ; Ejaculation ; Expressed Sequence Tags ; Gene Expression Profiling ; Humans ; Male ; RNA, Messenger ; genetics ; Spermatozoa ; physiology

OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.

DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.

STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.

RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.

CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.

Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family

Objective By comparing with the SD rats mixed glial cells, C57 mice mixed glial cells and Kunming mice mixed glial cells and exploring the expression of inflammatory factor of three mixed glial cells induced by lipopolysaccharide (LPS), the study aimed to explore the application value of three kinds of mice and find the ideal model of inflammation. Methods We used LPS as inducers, and NO, IL － 1β, COX－2and iNOS as anti－inflammatory antioxidant index. After cutting the head and taking the brain, we cultured the mixed glials. Then we used Greiss assay to detect the expression of NO and used western blot to detect the expression of protein of IL－1β, COX－2, and iNOS protein.Finally we compared the mixed glial cells from SD rats, C57 mice mixed glial cells and Kunming mouse mixed glial cells and selected the best inflammation model from three mixed glial cells. Results The results showed that the morphological changes of the mixed glial cells in SD rats were treated with N2－ free medium. Compared with the control group, the quantity of NO of LPS group of three mixed glial cells increased significantly (P＜0.01) . The LPS group of SD rats released the highest concentration of NO. Western blot was used to detect the expression of IL－1β, Cox－2 and iNOS in three kinds of rodents. Compared with the blank control group, the expression of COX－2 protein, iNOS and IL－1β in the LPS group of the three mice increased significantly. The results showed that LPS could successfully stimulate the release of inflammatory cytokines in three kinds of mice,among which the SD rats were more sensitive and it could be used in the study of AD inflammation model. Conclusion The results showed that LPS could induce the release of NO and the expression of IL－1β, iNOS and COX－2 in C57BL/6 mice,Kunming mice and SD rats to induce inflammatory response. Thus,LPS can induce the formation of inflammatory oxidation models of the original mixed glial cells of the three mice. Moreover, the SD rats were more sensitive.

AIM:To evaluate the effect of exogenous hydrogen sulfide(H2S)on the expression of NLRP3 in-flammasome in hepatocytes.METHODS:The hepatocytes L 02 and SMMC-7721 were used to establish the model of inflam-mation by stimulating with lipopolysaccharide(LPS)at different concentrations in vitro.The expression of NLRP3 inflam-masome in the hepatocytes was detected by Western blot and the cell viability was measured by MTT assay for determining appropriate concentration of LPS.The hepatocytes were divided into 4 groups:the cells in control group were incubated with normal medium for 18.5 h;the cells in LPS group were incubated with normal medium for 0.5 h followed by 100 μg/L LPS for 18 h;the cells in LPS+H2 S group and H 2 S group were incubated with 200μmol/L sodium hydrosulfide hydrate(NaHS)for 0.5 h followed by 100 μg/L LPS or normal medium for 18 h,respectively.The protein expression of NLRP3 and caspase-1 in the cells of every group was determined by Western blot.RESULTS:Compared with control group ,the protein expression of NLRP3 and caspase-1 increased significantly in LPS group(P<0.05)and had no significant change in H2S group.Compared with LPS group,the protein expression of NLRP3 and caspase-1 in LPS+H2S group decreased significantly(P<0.05).CONCLUSION:In hepatocytes,exogenous H2S suppresses the expression of NLRP3 inflamma-some.

OBJECTIVETo explore the possibility that the free latissimus dorsi musculo-cutaneous flap to repair the forearm leg wound.

METHODSTo design latissimus dorsi musculo-cutaneous flap which is foundation on T form thoracodorsal artery stalk. To set the short arm into the receiver artery break and anastomos them. It is not only reassure the blood of free musculo-cutaneous flap, but also reconstruct the continuation of the receiver main artery.

RESULTSIn 16 patients, 15 patients success completely, 1 patient main success. The blood supply of receiver is adequate.

CONCLUSIONSThe free T form thoracodorsal artery stalk musculo-cutaneous flap free grafting is a good method to repair the skin and soft tissues defection of forearm and leg.

Anastomosis, Surgical ; methods ; Arteries ; Forearm ; Free Tissue Flaps ; blood supply ; transplantation ; Humans ; Leg ; Lower Extremity ; Reconstructive Surgical Procedures ; Skin ; Superficial Back Muscles ; blood supply ; transplantation ; Wound Healing

BACKGROUNDIntegrase interactor 1 (INI1), which encodes a component of the ATP-dependent chromatin remodeling hSWI-SNF complex, has been identified as a tumor suppressor in many tumors. Nonetheless, the role of INI1 in gastric tumor progression is not known exactly. The aim of this research was to investigate the effect of INI1 in the carcinogenesis and progression of gastric cancer.

METHODSGastric tumor tissues with different differentiation levels from clinical gastric carcinoma samples and adjacent control normal tissues were taken. Expression levels of INI1 were detected by quantitative reverse transcriptation-polymerase chain reaction (RT-PCR) and Western blotting. Gastric cancer cell line SGC7901 was transfected with INI1 eukaryotic expressing vector INI1-GFP. Cell proliferation activities were assessed by MTT; cell count and cell cycle were detected by flow cytometry (FCM); cell apoptosis were measured by TUNEL and FCM; cell migration and invasiveness were evaluated by wound healing and transwell assays. Expression levels of INI1 and proliferation-related genes including p16, p21, cyclin D1 and cyclin A, apoptosis genes p53, B-cell non-Hodgkin lymphoma-2 (Bcl-2), Bcl-2-associated x protein (Bax) and caspase-3, and invasion-related genes including intercellular adhesion molecule 1 (ICAM1), matrix metalloproteinase 2 (MMP2), MMP9 and tissue inhibitor of matrix metalloproteinase 1 (TIMP1), were detected by quantitative RT-PCR and Western blotting.

RESULTSINI1 expression levels were lower in gastric carcinoma compared with adjacent control normal tissues. Overexpression of INI1 in SGC7901 cells inhibited its proliferation and invasiveness, but increased anoikis and G(0)/G(1) cell number. INI1-GFP transfection upregulated expression of INI1 and proliferation related genes p16 and p21, apoptosis genes p53 and Bax, and invasion-related genes TIMP1; cyclin D1, cyclin A, Bcl2, ICAM1, MMP2 and MMP9 were downregulated, and there was no significant change in caspase 3 levels.

CONCLUSIONINI1 plays a key role in gastric carcinogenesis by affecting proliferation, apoptosis and invasion.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Cycle ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Humans ; Real-Time Polymerase Chain Reaction ; SMARCB1 Protein ; Stomach Neoplasms ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo construct the prokaryotic and eukaryotic expression vectors pET32/E5 and pcDNA3.1/E5 for transformation into E. coli BL21 and NIH(3)T(3) cells respectively to observe the expression of human papillomavirus type 16 E5 protein (HPV16 E5).

METHODSHPV16 E5 gene was amplified by PCR from clinical isolates of HPV 16 and inserted into the plasmid pET32a(+) followed by digestion with BamH I and Hind III. The recombinant plasmid pET32/E5 was transformed into E. coli JM109 and selected with ampicillin. The positive clones containing the recombinant plasmid pET32/E5 were verified by restriction endonucleases BamH I and Xho and sequence analysis. The expression of HPV16 E5-TRX fusion protein in E. coli BL21(ED3) was identified by SDS-PAGE and Western blotting. The digestion product of BamH I and Xho was purified and inserted into the eukaryotic expression vector pcDNA3.1(+) to construct pcDNA3.1/E5, which was identified by sequencing and transfected into NIH3T3 cells. The NIH(3)T(3) cells with stable expression of HPV16 E5 were selected by G418 and confirmed by RT-PCR.

RESULTSThe pET32/E5 and pcDNA3.1/E5 vectors were constructed successfully. E.coli BL21(DE3) transformed by the recombinant plasmid pET32/E5 expressed HPV16 E5-TRX fusion protein efficiently. In the presence of 1 mmol/L IPTG at 28 degrees C, HPV16 E5-TRX recombinant protein accounted for about 10% of the total bacterial proteins. NIH3T3 cells stably expressing HPV16 E5 were harvested by selection with 250 g/ml of G418. HPV16 E5 gene from pcDNA3.1/E5-transfected NIH(3)T(3) cells was amplified by RT-PCR, and sequence analysis demonstrated the acquisition of the full-length gene fragment.

CONCLUSIONSThe prokaryotic and eukaryotic vectors for the HPV16 E5 gene have been successfully constructed. The acquisition of E .coli and NIH(3)T(3) cells with stable expression of HPV16 E5 protein may facilitate subsequent research of the biological properties and the transformation mechanism of HPV16 E5 protein on specific cells.

3T3 Cells ; Animals ; Escherichia coli ; genetics ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Mice ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the ways testosterone influences the murine bone marrow-derived macrophages (BMMs) and how testosterone affects the function of BMMs after bound to their membrane surface.

METHODSBMMs were cultured in vitro, their total RNA and proteins isolated, and the expression of intracellular androgen receptor (AR) detected through RT-PCR and Western blotting. The binding site of testosterone (T) to the membrane surface of BMMs was observed by confocal laser scanning microscopy after T-BSA-FITC incubation. Moreover, the intracellular Ca2+ was tested by Fura-2 method, and the influence of ionic currents on BMMs plasma membrane induced by testosterone was examined by the whole cell patch-clamp.

RESULTSRT-PCR and Western blotting failed to detect intracellular ARs in BMMs, but confocal laser scanning microscopy showed testosterone to be bound to the membrane surface of BMMs by impermeable T-BSA-FITC, inducing a rapid rise in the intracellular free Ca2+ concentration ([Ca2+]i) of Fura-2 loaded BMMs, predominantly due to the influx of extracellular Ca2+ through Ni2+ -blockable Ca2+ channels in the plasma membrane. Similarly, the patch-clamp technique revealed T-induced calcium influx in BMMs.

CONCLUSIONIt is reasonable to assume that the testosterone receptor exists on the plasma membranes, and testosterone act through unconventional plasma membrane receptors, induce Ca2+ influx and a rapid rise in the intracellular Ca2+ concentration, and influence the function of BMMs.

Animals ; Blotting, Western ; Calcium ; metabolism ; Calcium Channels ; physiology ; Cell Membrane ; metabolism ; Cells, Cultured ; Female ; Macrophages ; cytology ; metabolism ; physiology ; Membrane Potentials ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Protein Binding ; Receptors, Androgen ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; metabolism