OBJECTIVETo establish human colorectal tissue model in HIV-1 mucosal infection and by using pseudotyped virus to simulate the biological process of HIV-1 mucosal infection from HIV-1 entering into mucosa to local infection establishment.

METHODSTumor adjacent normal colorectal tissues were obtained with informed consent. After excised the muscularis externa, the mucosa and submucosa were dissected into the same blocks and cultured in 12-well cell culture plates. The cultured tissue structure and morphology were observed from day 0 to day 13 by staining with the hematoxylin eosin (HE), and the tissue activity was detected by 3(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The established tissues explants were infected by a single cycle replicated pseudotyped virus and propagated for 6 - 7 days, then subjected to the detection of p24 production within supernatant to verify the applicability of the model for the studying of HIV-1 mucosal infection. The applicability of the established explants for safety and reactivity evaluation of mucosa topical drugs was conducted by the using of first generation antiseptic Nonoxynol-9 (N-9) as an example.

RESULTSHE staining showed the structure of colorectal tissue was remained well until 5(th) day and still evident until 13(th) day. The tissue activity of cultured mucosa was above 80% at day 4, and still remained over 50% at day 7 as detected by MTT assay. After infected by pseudo virus, the increased level of p24 was detected from supernatant collected on 1(st), 4(th), 8(th) day, which indicated a local infection was created. In addition, the dose changing of N-9 was reflected sensitively by the activity of this model.

CONCLUSIONEx vivo human colorectal tissue model mimic HIV-1 mucosal infection was established that can be used to replicate the bioprocess of human HIV-1 mucosal infection.

Colon ; pathology ; virology ; HIV Infections ; pathology ; virology ; HIV-1 ; Humans ; Intestinal Mucosa ; pathology ; virology ; Models, Biological ; Rectum ; pathology ; virology ; Tissue Culture Techniques ; methods ; Tumor Cells, Cultured

We report a case of a 72-year-old woman with Churg-Strauss syndrome, who presented with intestinal perforation. She has had bronchial asthma with peripheral blood eosinophilia for 30 years. Gross findings of a resected colon showed multiple ulcers with perforation. Histologic findings demonstrated transmural inflammation infiltrated with large numbers of eosionophils, neutrophils and lymphoplasma cells, and characteristic extravascular granuloma in the subserosa. There were multifocally-distributed transmural vasculitis showing all stages of activity in medium and small-sized arteries and veins located in the submucosa, and proper muscle and subserosal layers of the colon, some of which revealed granulomatous inflammation. Histologic finding of liver showed chronic viral hepatitis B with mild inflammatory activity and macronodular cirrhosis. Immunohistochemical findings, acid fuschin orange G staining and electromicroscope found no evidence of hepatitis B virus infection contributing to the pathogenesis of this lesion.
Aged ; Case Report ; Churg-Strauss Syndrome/virology ; Churg-Strauss Syndrome/pathology ; Churg-Strauss Syndrome/complications* ; Colon/virology ; Colon/pathology* ; Colonic Diseases/virology ; Colonic Diseases/pathology ; Colonic Diseases/etiology* ; Female ; Hepatitis B/pathology ; Hepatitis B Antigens/analysis ; Human ; Immunohistochemistry ; Intestinal Perforation/virology ; Intestinal Perforation/pathology ; Intestinal Perforation/etiology*

Perforation of the colon occurs in 0.2 to 2% of all colonoscopic examinations. The most common sites of perforation are rectosigmoid junction and cecal area. Colonic perforation, leading to tension pneumoperitoneum in most cases, may be caused by direct trauma or pressurized air. It should be suspected in patients with hypotension, tachycardia and tachypnea during or after the colonoscopy. An 83-year-old woman was admitted due to pulmonary embolism and left cerebellar infarction. Colonoscopy was performed due to bloody diarrhea. She was diagnosed as cytomegalovirus (CMV) colitis. One week after the colonoscopy, colon perforation was incidentally found on ascending colon, and tension pneumoperitoneum occurred immediately after the procedure. The perforated site was primarily closed and the patient discharged 20 days later. Herein, we report a case of tension pneumoperitoneum following colonoscopy in a patient with CMV colitis.
Aged, 80 and over ; Colitis/*diagnosis/virology ; Colon/*injuries ; Colonoscopy/*adverse effects ; Cytomegalovirus Infections/*diagnosis ; Female ; Humans ; Intestinal Perforation/*etiology ; Pneumoperitoneum/*etiology

UNLABELLEDTo provide a reliable animal model for study of human CMV disease in gastrointestinal track, we tried to infect with murine cytomegalovirus (MCMV) in mice that were received allogenetic skin transplantation under immunosuppression. (1) Skin transplantation was performed between 18 donor C57BL/6 mice and 72 recipient BALB/c mice. (2) All recipient mice were then given Cyclosporine at 12 mg/kg daily for 2 weeks by intraperitoneal injection. Mice were randomly divided into 3 groups. Two experimental groups were received MCMV-infected mouse embryonic fibroblasts (MEF) at 10(4) PFU and 10(5) PFU respectively, and the control group received MEF only. We observed any possibly pathophysiological behavior changes and recorded the changes in body weight. The mice were sacrificed at 5d, 9d, 14d, 21d post infection and colon tissue was collected for analysis.

RESULTSMice infected with MCMV at 10(5) PFU group showed anorexia, lethargy and degression in locomotor activity. This group of mice showed significant decrease in body weight than that of other groups. Colon tissues were collected 14 days after infection. Histological examination revealed that the mucous layer became thinner in the proximal colon and increased number of lymphoid follicles in distal colon in infected animals. The changes in the mucosal structure was most prominent in the group 10(5) PFU MCMV. Viral DNA was present in the colon by in situ hybridization for IE1 gene, and viral gB transcript was positive by RT-PCR. One of the viral major proteins, pp65, was widely distributed in the colon by immunohistochemistry. These data demonstrated that MCMV established infection in colon of the mice after allogenetic skin transplantation. Electron microscopy showed that there were herpes virus particles in the colon tissue.

CONCLUSIONInfection with MCMV in mouse after allogenetic skin transplantation by nasal cavity inoculation resulted in the pathological changes in colon tissue similar to that of inflammation in human colon. The small animal model of colon inflammation may provide a platform for further study of pathogenesis as well as medical intervention of HCMV involved inflammation of human bowel.

Animals ; Colon ; immunology ; pathology ; virology ; Cytomegalovirus Infections ; immunology ; pathology ; virology ; Disease Models, Animal ; Female ; Herpesviridae Infections ; immunology ; pathology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Muromegalovirus ; genetics ; immunology ; isolation & purification ; Random Allocation ; Skin Transplantation ; adverse effects ; immunology ; pathology ; Transplantation, Homologous ; adverse effects ; immunology ; pathology ; Viral Proteins ; genetics ; metabolism