Objective To investigate the expression and effect of secondary lymphoid tissue chemokine(SLC)in experimental ulcerative colitis(UC)in rats.Methods Thirty Spague-Dawley rats were divided into control group and UC group.SLC expression in colon tissues was detected by RT-PCR and immunohistochemistry,respectively.Results The transcriptional level of SLC in UC tissues was significantly higher than that in the control group(0.846?0.07 vs 0.312?0.12,P＜0.01).The positive expression of SLC was concentrated mainly on submucosa,and the positive rate of SLC protein expression in UC group significantly higher than that in control group(P＜0.01).Conclusion SLC overexpression could contribute to the pathological processes in UC rats,thus SLC may be an ideal ther- apeutic target for UC.

OBJECTIVETo investigate the role of integrin α4β7 in the development of ulcerative colitis (UC) in rats.

METHODSSixty Sprague-Dawley rats were randomly divided into the control group (acetone enema), the model group (2,4-dinitrochlorobenzene, DNCB enema), and the α4 intervention group. Colonic mucosa of different groups was observed and compared in terms of pathology and cytokine changes(IL-2 and IL-6) using ELISA. Semi-quantitative RT-PCR was used to detect the colon α4β7 expression. Integrin α4β7(+) lymphocytes in the portal vein of rats were determined by flow cytometry.

RESULTSThe expression of α4 mRNA was 0.68±0.24 in the model group and 0.58±0.37 in the intervention group, and the expression of β7 mRNA was 0.84±0.37 in the model group and 0.65±0.30 in the intervention group, which were all significantly higher as compared to those in the control group(0.15±0.13 for α4 and 0.24±0.62 for β7, P<0.01). The proportions of integrin α4β7 positive lymphocytes in the portal vein in the model group and intervention group were significantly higher than that in the control group [(76.7±8.2)% and (68.2±7.6)% vs. (14.7±6.7)%, P<0.01]. The expression of IL-2 and IL-6 and the result of macroscopic and microscopic scores in the intervention group were lower than those in the model group(P<0.05).

CONCLUSIONSHigh expression of α4β7 may play an important role in experimental colon mucosa inflammation in rats with ulcerative colitis. The blockade of integrin α4β7 may be a potential target to reduce colonic mucosa inflammation.

Animals ; Colitis, Ulcerative ; metabolism ; pathology ; Colon ; metabolism ; pathology ; Disease Models, Animal ; Female ; Integrins ; metabolism ; physiology ; Interleukin-2 ; metabolism ; Interleukin-6 ; metabolism ; Intestinal Mucosa ; pathology ; Rats

Adult ; Chylous Ascites ; therapy ; Female ; Fibrin Tissue Adhesive ; therapeutic use ; Humans ; Middle Aged ; Parenteral Nutrition, Total ; Somatostatin ; therapeutic use

OBJECTIVETo investigate the effect of secondary lymphoid tissue chemokine (SLC) on experimental colon lesions in rats with ulcerative colitis.

METHODSSixty Sprague-Dawley rats were randomly divided into control group, model group and SLC intervention group. Colonic mucosal lesions of different groups were observed with HE staining for inflammation and lymphocyte homing situation. Cytokine IL-2 and IL-6 levels were measured by ABC-ELISA. Semi-quantitative RT-PCR was used to examine the colonic SLC expression.

RESULTSIntestinal inflammation score and colonic cytokine levels were significantly different among three groups (P<0.05, P<0.01). Abnormal lymphocyte homing phenomenon under colonic mucosa was found in the model group and the intervention group. SLC mRNA expression of the model and intervention groups increased significantly compared with the control group (0.846+/-0.047, 0.768+/-0.135 vs 0.312+/-0.112, P<0.01). However, there was no significant difference between model group and intervention group.

CONCLUSIONSSLC may play an important role in experimental colonic mucosal inflammation in rats with ulcerative colitis. Blockade of SLC may be one of effective ways in reducing colonic mucosal inflammation.

Animals ; Chemokine CCL21 ; metabolism ; Colitis, Ulcerative ; metabolism ; physiopathology ; Female ; Inflammation ; Interleukin-2 ; metabolism ; Interleukin-6 ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the clinical application of aoptimizedtechniquebased onpreviouslyreported protecting stoma with no need forreversal.

METHODSThetechniquealso used "the assembly of drainage device" to performprotecting ileostomy. The original method includes enterotomy at the terminal ileum to placedrainage device, which was optimized as follows: two intestinal pursestring with 0.5 cm distance were placed 5 cm away from the ileocecal valve. Transverse enterotomy was performed in the anti-mesenteric side. The assembly was placed at the root of the appendix between two pursestring, and then the intestine purse suture was tighten. Ligation of the small intestine anastomosis between the anastomosis ring at both ends was carried out, and theanastomosis ring was deployed. From the root of the appendix in the cecum wall, the assembly was embedded about 2 cm and pulled out of abdominal cavitythough the Trocar hole.

RESULTSSeventeen cases of ultra-low rectal cancer completed protecting stoma, including 11 cases through ileocecal protective stoma. All the anastomosis healed well. Defecation drainage tube was removed 3-5 weeks after anastomosis ring degradation. Drainage nozzle healed after 3 to 5 days, and no complications occurred.

CONCLUSIONThe optimized ileocecal protective ileostomy has the following advantages: (1)wound healing time is significantly shorter. (2)secondary intestinal fistula can be prevented. (3)no need to fix ileum and less chance of subsequent volvulus, intestinal obstruction.

Anastomosis, Surgical ; Defecation ; Drainage ; Humans ; Ileostomy ; methods ; Ileum ; surgery ; Intestinal Fistula ; Rectal Neoplasms ; Surgical Stomas

In this study, Recombinant baculoviruses rBac-NF and rBac-NG were generated for expressing F and G proteins Nipah virus (NiV) . The expression of recommbinant G (rNG) and F (rNF) protein in rBac-NF and rBac-NG infected cells were confirmed by western-blot. Both rNG and rNF showed sensitive and specific antigenic reaction to rabbit serum anti-Nipah virus in indirect immunofluorescence detection and indirect ELISA. Immunization with rBac-NF and rBac-NG infected insect cells elicited G ad F protein specific antibody responses in mice. Furthermore, the G ad F specific antibodies could neutralize the infectivity of the VSVdeltaG* F/G, the NiV F and G envelope glycoproteins psudotyped recombinant Vesicular Stomatitis Virus expressing green fluorescence protein. The results demonstrated F and G protein expressed by the recombinant baculoviruses could be safe economic diagnostic antigens for the surveillance and monitoring of NiV and promising subunit vaccines for the prevention of NiV.
Animals ; Antigens, Viral ; immunology ; Baculoviridae ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Nipah Virus ; chemistry ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Recombination, Genetic ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology

The full-length porcine interferon gamma(PoIFN-gamma) cDNA, including the secretion signal peptide coding region was recloned into honor plasmid pFastBac 1 of Bac-To-Bac Baculovirus Expression System. These recombinant plasmids, pFastBac -PoIFN-gamma, were transformed into DH(10Bac) host bacteria to get recombinant shuttle plasmids, rBacmid-PoIFN-gamma. Recombinant baculovirus, rBac-PoIFN-gamma, was generated for expressing PoIFN-gamma, by transfecting rBacmid-PoIFN-gamma with Cellfectin Reagent into sf9 insect cells. The expression of PoIFN-gamma in insect cells was confirmed by Western Blot, indirect immunofluorescence assay and indirect ELISA. The antiviral activity assay shows that PoIFN-gamma expressed by the rBac-PoIFN-gamma can efficiently inhibit the replication of the recombinant Vesicular Stomatitis Virus expressing green fluorescence protein in PK-15 cells. The antiviral activity of PoIFN-gamma can be specifically blocked by anti-PoIFN-gamma mouse serum. The antiviral titer of culture supernatant of insect cells infected by rBac-PoIFN-gamma is 2 x 10(4) IU/mL. The results demonstrat that the rBac-PoIFN-gamma can express rPoIFN-gamma efficiently and rPoIFN-gamma has high antiviral activity.
Animals ; Antiviral Agents ; metabolism ; pharmacology ; Baculoviridae ; genetics ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Green Fluorescent Proteins ; genetics ; metabolism ; Immune Sera ; immunology ; Interferon-gamma ; genetics ; immunology ; metabolism ; pharmacology ; Mice ; Mice, Inbred BALB C ; Microscopy, Fluorescence ; Recombinant Proteins ; Spodoptera ; Swine ; Vesiculovirus ; genetics ; Virus Replication ; drug effects

The two mammalian codon optimized genes, F and G genes of Nipah virus, were generated by assembly PCR, and inserted into mammalian expression vector pCAGGS under chicken beta-actin promoter to construct pCAGG-NiV-F and pCAGG-NiV-G. Syncytium formation was induced in BHK cells by plasmid pCAGG-NiV-F and pCAGG-NiV-G transfection, which indicate recombination proteins F and G were expressed in BHK cell and possessed good biologic activity. Six-week-old female BALB/c mice were intramuscularly primed with 100 microg pCAGG-NiV-F, pCAGG-NiV-G or pCAGG-NiV-F+ pCAGG-NiV-G respectively, and boosted with same dose after 4 weeks. The sera were collected at 3 weeks post second boost. The serum IgG against Nipah virus F and G proteins was detected by indirect ELISA using recombinant Baculovirus expressed Nipah F and G glycoproteins. The results showed that specific antibodies possessed good sensitivity and specificity. Furthermore, the G and F proteins' specific antibodies could neutralize the infectivity of VSVdeltaG* F/G (the NiV F and G envelope glycoproteins psudotyped recombinant vesicular stomatitis virus expressing green fluorescence protein). And, pCAGG-NiV-G also induced higher titer of neutralizing antibody response than pCAGG-NiV-F did. The result indicates that DAN immunization is an efficient vaccine strategy against Nipah virus.
Animals ; Antibodies, Viral ; blood ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Female ; Mice ; Mice, Inbred BALB C ; Nipah Virus ; immunology ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Vaccines ; immunology

OBJECTIVETo study the expression and significance of GCS gene in human pancreatic cancer cell line SW1990 and its drug-resistant sublines.

METHODSSW1990 and its drug-resistant sublines, SW1990/FU, SW1990/ADM and SW1990/GEM were cultured in vitro. CCK-8 (Cell Counting kit-8) was used to detect the drug resistance of the sublines. Relative quantitation of GCS mRNA expression was evaluated by real-time PCR and Western blot was adopted to evaluate the expression of GCS protein.

RESULTSThe drug resistance indexes of SW1990/FU, SW1990/ADM and SW1990/GEM were 339.7, 11.9 and 56.6, respectively. The GCS mRNA and protein were expressed in all SW1990 and its drug-resistant sublines. There was a higher expression of GCS mRNA in all the sublines and a significant difference of GCS protein expression was detected in SW1990/ADM and SW1990/GEM compared with SW1990.

CONCLUSIONSGCS gene is expressed in SW1990 and its drug-resistance sublines. The high expression of GCS protein in SW1990/ADM and SW1990/GEM might be one reason of resistance to ADM and GEM in the sublines.

Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Gene Expression ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Immunoblotting ; Pancreatic Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Rabies remains an important worldwide health problem. Newcastle disease virus (NDV) was developed as a vaccine vector in animals by using a reverse genetics approach. Previously, our group generated a recombinant NDV (LaSota strain) expressing the complete rabies virus G protein (RVG), named rL-RVG. In this study, we constructed the variant rL-RVGTM, which expresses a chimeric rabies virus G protein (RVGTM) containing the ectodomain of RVG and the transmembrane domain (TM) and a cytoplasmic tail (CT) from the NDV fusion glycoprotein to study the function of RVG's TM and CT. The RVGTM did not detectably incorporate into NDV virions, though it was abundantly expressed at the surface of infected BHK-21 cells. Both rL-RVG and rL-RVGTM induced similar levels of NDV virus-neutralizing antibody (VNA) after initial and secondary vaccination in mice, whereas rabies VNA induction by rL-RVGTM was markedly lower than that induced by rL-RVG. Though rL-RVG could spread from cell to cell like that in rabies virus, rL-RVGTM lost this ability and spread in a manner similar to the parental NDV. Our data suggest that the TM and CT of RVG are essential for its incorporation into NDV virions and for spreading of the recombinant virus from the initially infected cells to surrounding cells.
Animals ; Antibody Formation ; Cytoplasm* ; Glycoproteins* ; GTP-Binding Proteins ; Humans ; Mice ; Newcastle disease virus* ; Newcastle Disease* ; Parents ; Rabies virus ; Rabies* ; Reverse Genetics ; Tail* ; Vaccination ; Virion*