Effects of intestinal microbiota of patients with chronic constipation on the expression of serotonin transporter and the bowel movement in mice / 中华消化杂志

Objective To investigate the effects of intestinal microbiota of patients with chronic constipation on the expression of serotonin transporter (SERT) and the bowel movement in mice.Methods Fecal samples of patients with slow transit constipation met Rome Ⅲ criteria and healthy normal controls were collected and made into fecal microbiota solution.Twenty specfic pathogen free (SPF) mice were divided into experiment group and control group.The mice of two groups were both pre-treated with streptomycin to establish the germ-free mice model.The mice of control group were gavaged with mixed fecal microbiota solution of healthy normal controls and the mice of experiment group were gavaged with mixed fecal microbiota solution of patients with chronic constipation.Mice were sacrificed after fed for 15 days.Defecation parameters and ink discharge time of mice were detected.The expressions of SERT mRNA and SERT protein in mice intestinal tissues were detected with real time-polymerase chain reaction and Western blotting.The 5-hydroxytryptamine (5-HT) levels of mice intestinal tissues were evaluated by enzyme-linked immunosorbent assay (ELISA) and double immunofluorescent staining.The methods of t test and Chi-square test were performed for statistical analysis.Results On the 15th day,the total number of the feces within 2 h of the experiment group and control group was 8.55±1.83 and 12.14±2.90,respectively,and the difference was statistically significant (t=3.33,P＜0.05).The weight of feces were (151.90 ± 32.42) mg and (246.72 ± 64.01) mg,respectively,and the difference was statistically significant (t=4.18,P＜0.01).The dry weight of feces were (65.52±11.76) mg and (92.93±23.07) mg,respectively,and the difference was statistically significant (t=3.37,P＜0.05).The water content of feces were (56.63 ± 3.01) ％ and (61.95 ± 3.70) ％,respectively,and the difference was statistically significant (t=3.57,P＜0.05).The defecating time of first black feces of the experiment group and control group were (83.24±11.31) min and (69.06±2.72) min,respectively,and the difference was statistically significant (t=-2.74,P＜0.05).The expressions of SERT mRNA and SERT protein levels in mice intestine tissues of the experiment group were significantly higher than those of the control group (t =2.61,-6.89;both P＜0.05).5-HT level of mice intestinal tissues of the experimental group and control group were (151.69± 10.18) ng/mL and (198.77 ± 25.99) ng/mL,respectively,and the difference was statistically significant (t=-4.13,P＜0.01).Conclusion Intestinal microbiota of patients with chronic constipation may influence the expression of SERT in the mice intestinal tissues,and then decrease the level of 5-HT,slowing the bowel movement in mice.

Surgery for ulcerative colitis in the era of biological agents therapy / 中华消化外科杂志

Ulcerative colitis is an inflammatory disease of colon and rectum whose etiology is still unclear.Infliximab is an anti-tumor necrosis factor antibody,which has been approved recently by the United States FDA for the treatment of ulcerative colitis to reduce signs and symptoms,to induce clinical remission and healing of the intestinal mucosa.Total proctocolectomy with pouch-anal anastomosis are the standard operation for ulcerative colitis now.The perioperative infliximab use,operation timing and procedures are the important factors affecting prognosis of patients in the era of infliximab therapy.

Producing and quality control of fecal-derived microbiota enteric capsules / 中华消化杂志

Objective To initially explore the feasibility and quality control of producing fecal-derived microbiota enteric capsules.Methods Fecal-derived microbiota was put into double layered enteric capsules.The bacteria colony numbers of fresh prepared glycerol containing fecal-derived microbiota liquid,glycerol containing and glycerol free fecal-derived microbiota after stored at -80 ℃ for 72 h were counted with standard plate count methods in order to investigate the stability after frozen.Methylene blue was taken as the standard,resistance to acid and release rate of capsules was evaluated.The t test was performed for statistical analysis.Results The preparation process of double layered microbiota capsules was simple and practicable.The data of 12 plates of each microbiota were acquired.The number of bacteria colony of fresh prepared glycerol containing fecal-derived microbiota ((5.08 ±1.37)×107 colony-forming units (cfu)/mL)was significantly more than that of the group without glycerin ((1.73±0.64)×107 cfu/mL)at -80 ℃for 72 h (t = 7.621 ,P <0.01).There was no significant difference in the number of bacteria colony between glycerin containing frozen fecal-derived microbiota ((4.67±1 .56)×10 7 cfu/mL)and fresh fecal-derived microbiota (t = 0.694,P = 0.495).Regression equations were achieved with fecal-derived microbiota containing methylene blue,and there was a good linear relation between 0.5 μg/mL and 8.0 μg/mL.Three fecal-derived microbiota enteric capsules containing methylene blue were prepared.Their resistance to acid was 96.0%,99.1 %,and 95 .5 %,while the release rate was 88.6%,95 .1 % and 86.5 %, respectively.All met the requirement of Chinese Pharmacopoeia to enteric capsules.Conclusion The preparation of double layered fecal-derived microbiota enteric capsules had feasible technology and stable quality,which could provide reference in prevention and treatment of diseases related with colonic microbiota imbalance.

Secondary bile acid induced intestinal adenoma canceration and its effect on intestinal microflora in Apcmin/+ mice / 中华消化杂志

Objective To investigate secondary bile acid induced canceration process of intestinal adenoma and effects on intestinal microflora in Apcmin/+ mice.Methods Forty four-week-old mice (20 Apcmin/+mice and 20 wild-type C57BL/6J mice) were divided into four groups:wild-type control group (regular drinking water),wild-type deoxycholic acid (DOC) group (with 0.2 ％ DOC in drinking water),Apcmin/+ control group and Apcmin/+ DOC group.Fecal pellets of Apcmin/+ mice were collected at 0 week and 12 week after administration.The changes of intestinal microflora were analyzed by pyrosequencing.All mice were sacrificed after 12 weeks.The number,size and location of intestinal adenoma were observed.The pathological type of adenoma was evaluated after hematcxylin-eosin (HE) staining.Proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry.Cell apoptosis was determined by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling technique (TUNEL).Independent t test was used for the quantitative data comparison between two groups.Results No intestinal tumors were found in the wild-type mice.The total number of intestinal adenoma of Apcmin/+ DOC group significantly increased,compared with that of Apcmin/+ control group (57.00 ± 3.07 vs 21.50± 4.69,t=20.03,P＜0.01),the increase of the adenoma with maximum diameter between 1 to 2 mm was most significant (30.62± 7.73 vs 7.75 ± 4.59,t =8.04,P＜ 0.05),the rate of adenoma canceration also significantly increased compared with that of Apcmin/+ control group.The percentage of PCNA positive cells significantly increased compared with that of Apcmin/+ control group ((90.17 ± 2.14) ％ vs (41.97 ± 4.26) ％,t=31.97,P＜0.01).The percentage of cell apoptosis significantly declined ((1.40± 1.12) ％ vs (7.50 ± 0.65)％,t =14.90,P＜ 0.01).The diversity of intestinal flora of Apcmin/+ DOC group significantly decreased.The ratio of Firmicutes and Bacteroidetes significantly increased compared with control group (0.586 7±0.148 4 vs 0.387 3±0.013 6,t=2.36,P＜0.05).The number of pathogenic bacteria increased in Apcmin/+ DOC group and probiotics significantly decreased.Conclusion DOC can induce intestinal flora imbalance in Apcmin/+ mice and promote intestinal adenoma into adenocarcinoma through increasing tumor cell proliferation and inhibiting cell apoptosis.

Expression of CDX2, COX-2, and NF-κB in gastric carcinoma and precancerous lesions / 中国肿瘤临床

Objective: To detect the expression of CDX2, COX-2, and NF-κB in the tissues of gastric carcinoma (GC), precancerous lesions, and normal gastric mucosa and to in vestigate the correlation among CDX2, COX-2, and NF-κB. Methods:Data on the expression of CDX2, COX-2, and NF-κB were evaluated using immunohistochemistry in 60 cases with GC, 45 cases with precancerous lesions, and 20 cases with normal tissues. The protein levels in different pathological tissues, as well as their correlation with GC, were analyzed. Results:The positive rates of NF-κB and COX-2 proteins were significantly higher in GC group than those in precancerous lesion and normal tissue groups (P<0.05). CDX2 was expressed in GC and precancerous lesion groups but poorly expressed in the normal mucosa. The positive rate of CDX2 expression was significantly higher in precancerous lesion group than that in GC group (P<0.01). NF-κB expression was positively correlated with COX-2 in GC (P<0.01), whereas CDX2 expression was negatively correlated with COX-2 and NF-κB in GC (P<0.01). Conclusion:CDX2, COX-2, and NF-κB are possibly correlated with one another, and they cooperate during the onset, progression, and metastasis of GC.

Color Doppler ultrasonography in assessment of activity of ulcerative colitis / 中华全科医师杂志

Sixty patients with ulcerative colitis (UC),including 28 males and 32 females were enrolled in the study.Color Doppler ultrasonographic scans were performed and the disease activity was estimated base on the Sourtherland score index (DAI).On the sonography the colon with active inflammatory disease showed bowel wall thickening,alteration of layers structure or even discrimination and increased vascular signals.Using DAI as the gold standard,the sensitivity,specificity,positive predictive value,negative predictive value and accurate rate for ultrasonography in assessment of UC activity were 90.0％ (36/40),75.0％ ( 15/20),87.8％ (36/41),78.9％ (15/19),85.0％ (51/60),respectively.In conclusion,Color Doppler ultrasonography provides a convenient technique for assessment of bowel involvement and inflammatory activity in ulcerative colitis with high sensitivity and accuracy.

DNA damage of gastric cancer cell line SGC7901 dyed by methylene blue after exposure to the cold light radiation of endoscope / 中华放射医学与防护杂志

Objective To study the co-effects of methylene blue(MB) and exposure with the cold light source on cell DNA damage, and to explore the mechanism involved. Methods The alkaline single cell gel electrophoresis was used to determine cell DNA damage. Apoptosis of the cells was determined by flow cytometry analysis using Annexin V-FITC/PI staining. DCFH-DA probe was used to determine endocellular Reactive Oxygen Species (ROS). Results The levels of DNA damage in the SGC7901 adenocarcinoma cells treated with methylene blue in the light were significantly increased compared to that of control ( F = 8.39, P＜0.05 ). The DNA damage levels were related to the length of time of light exposure, and the damage was recovered to a certain level after light withdrew. Cell apoptosis ( x2=7.71,P ＜0.05)and endocellular ROS level (F = 34.11, P＜0.01＝ increased significantly in the exposure group. Conclusions Methylene blue chromoendoscopy can induce DNA damage and cell apoptosis, and the mechanism may be associated with ROS produced by the photochemical reaction.

A preliminary study on the relationship of insulin-like growth factor-Ⅰ receptor and the carcinogenesis of gastde cancer / 中华消化杂志

Objective To study the relationship of insulin-like growth factor-Ⅰ receptor(IGF-ⅠR)and carcinogenesis of gastric cancer.Methods The expression of IGF-ⅠR was detected in 40 cases of resected gastric cancer tissues and adjacent normal gastric mucosa by immunohistochemistry.The expression of IGF-ⅠR in gastric cancer cell lines(MGC803 and SGC7901)was detected by Western Blot.siRNAs targeted to IGF-ⅠR were designed,synthesized and transfected into MGC803 cells,the changes of IGF-IR protein level were detected by Western Blot at 48 h after transfection,the cell proliferation was examined by MTT,and then the growth curve was obtained.Results The positive rate of IGF-IR expression in gastric cancer tissues was 75.5%,significantly higher than that of adjacent normal tissues (25%,P＜0.01).The expression level of IGF-IR was related with TNM stage,lymphnode metastasis (P＜0.05),but not related with sex,age,histological differentiation,invasion depth of gastric cancer (P＞0.05).Intense expression of IGF-ⅠR was showed in gastric cancer cell lines.The inhibition ratio of IGF-ⅠR expression in sigNAl group were 89.80%±4.10%,the cell proliferation decreased to mininlunl level at the fifth day aftertransfection(by 29.0%±4.0%of mock-treated group),the cell number decreased by 21.15%±1.10%of mock treated group at the same time.Conclusions IGF-ⅠR is over-expressed in gastric cancer cells and can be effectively silenced by RNA interferes,therefore the growth of tmnor cell Was inhibited.Thus,it indicates that IGF-ⅠR may be a promising target for gene therapy of gastric cancer.

The utility of capsule endoscopy in the diagnosis of obscure gastrointestinal bleeding of the elderly / 中华老年医学杂志

Objective To evaluate the effectiveness of M2A capsule endoscopy in the diagnosis of obscure gastrointestinal bleeding of elder patients. Methods Capsule endoscopy was used in 27 elderly patients, who were suspected of small bowel bleeding because of obscure gastrointestinal bleeding. About 20 of the 27 patients underwent gastroscopy, colonoscopy or air-baricum double contrast examination, but none of the examinations confirmed the bleeding sites, whereas the other 5 patients didn't receive any examination because of their intolerance. Results M2A capsule endoscopy disclosed the sites of small bowel disorders in 20 out of the 27 patients, the positive rate was about 74. 1%. The pathological findings consisted of angiodysplasia in 6 cases, mutipolypi in the intestine in 4, and 1 of them was diagnosed rare Cronkhite-Canada syndrome, prominency in the intestine in 4, single or multiple ulcers in the intestine in 7, diffuse mucosa lesion in the intestine in 9 and diverticulum in 1, arteriolar bleeding in gastric in 2, gastric mucosa erosion or ulcer in 5, whereas 7 patients were in negative findings. There were 12 patients in whom appeared at least 2 sites of pathological changes. Conclusions This study demonstrates that capsule endoscopy provides an excellent visualization way for the small intestines, especially when the elder patients suffers obscure gastrointestinal bleeding. It is safe and usefull for evaluating the suspected small bowel bleeding.

The effect of Htr- antisense oligodeoxynucleotide in combination with DDP on human gastric carcinoma transplanted subcutaneously in nude mice in vivo / 肿瘤研究与临床

Objective To study whether hTR antisense PS ODN has anticancer effect and the effect of increasing the susceptibility of tumor to DDP, we explore the effect of hTR antisense oligodeoxynucleotide in combination with DDP on human gastric carcinoma transplanted subcutaneously in nude mice in vivo. Methods The model of human gastric carcinoma transplanted subcutaneously was established in thirty nude mice, then divided randomly into 5 groups (Control group, ASODN group, RODN group, DDP group and ASODN+DDP group). The weight of nude mice was measured, and the tumor growth inhibitory rate was calculated. The relative telomerase activity was quantitatively measured by TRAP PCR ELISA methods. Results The maximum tumor inhibitory rate in ASODN+DDP group, ASODN group, DDP group and RODN group was 94.2 %, 84.3 %, 92.8 % and 26.9 % respectively. There was significant difference in the relative telomerase activity among four treatment groups. Conclusions The results suggested that hTR antisense PS ODN may act as a specific tumor growth inhibitor and telomerase activity inhibitor may be in sequence specific manner, and have the effect of increasing the susceptibility of transplanted tumor to DDP.

Study on transplantation of adult stem cells to rat lower esophagus: a potential treatment for gastroesophageal reflux disease / 中华消化杂志

0. 05). No obvious local inflammation was found. Conclusions The study proved the feasibility and persistence of injecting SMSC and BMMSC into the lower esophagus. Transplantation of adult stem cells may serve as a potential new treatment for gastroesophageal reflux disease.

The effect of hTR antisense PS-ODN in combination with cisplatin and adriamycin on human gastric cancer cell line / 中华消化杂志

Objective To study the effect of hTR antisense PS-ODN on the susceptibility of human gastric cancer cell line SGC7901 to Adriamycin and Cisplatin. Methods hTR antisense PS-ODN with sequence CAG-TTAGGGTTAG which can recognize the RNA template of telomerase was transfected into SGC7901 by lipofectamine, and drug sensitivity test was performed with MTT method, telomerase activity was quantitatively measured by TRAP-PCR-ELISA methods, the apoptotic cells were measured with FCM. Results The telomerase activity in gastric cancer cell line SGC7901 after treatment with cisplatin and doxorubicin was different, and the change of telomerase activity of SGC7901 by cisplatin and doxorubicin was in time-dependent manner. The inhibitory ability of cisplatin and doxorubicin combined with hTR antisense PS-ODN in cell line SGC7901 was higher than that of these drugs used alone. Telomerase activity was dramatically declined during the cell incubation with hTR antisense PS-ODN combined with ADM or DDP. The hTR antisense PS-ODN increased susceptibility to cisplatin or doxorubicin-induced apoptotic cell death in gastric cancer cell line SGC7901. Conclusions The hTR antisense PS-ODN increased susceptibility to cisplatin or doxorubicin in gastric cancer cell line. This may be correlated with its inhibitory effects on telomerase activity and the ability of induced apoptotic cell death. These findings indicate that hTR antisense PS-ODN may represent a novel chemosensitive agent.