Objective To observe the effects of Huosha Oral Liquid on irritable bowel syndrome with diarrhea(IBS-D)rats with cold and dampness syndrome based on Notch1/Hes-1/Math-1 signaling pathway;To elucidate its mechanism of repair of intestinal mucosal barrier.Methods An IBS-D cold and dampness syndrome rat model was established by using restraint stress combined with bitter-cold cathartic method and cold and dampness environmental method.After modeling,the rats were randomly divided into model group,Western medicine group and Huosha Oral Liquid low-,medium-and high-dosage groups,with 6 rats in each group.An additional 6 rats were set as the normal group.The Western medicine group was given mebeverine(13.5 mg/kg)by gavage,while the Huosha Oral Liquid low-,medium-and high-dosage groups were given Huosha Oral Liquid(3.15,6.3,12.6 g/kg)by gavage,the normal group and model group were given distilled water by gavage,for 4 weeks.The general status of rats were ovserved,the body mass,loose stool rate,fecal water rate and abdominal withdrawal reflex(AWR)score were measured,the morphalogy of colon tissue were observed by HE staining,the mRNA expression of Occludin,ZO-1,Notch1,Hes-1 and Math-1 in colon tissue were detected by RT-PCR,the protein expression of Notch1 was detected by Western blot.Results Compared with the normal group,the model group had a reduced body mass(P<0.05)and increased loose stool rate,fecal water rate and AWR score(P<0.05),inflammatory cell infiltration was observed in colon tissue and the expressions of Occludin,ZO-1,Math-1 mRNA decreased(P<0.05),the expressions of Notch1,Hes-1 mRNA increased(P<0.05),and the expression of Notch1 protein increased(P<0.05).Compared with the model group,the body mass of rats in Western medicine group and Huosha Oral Liquid low-,medium-and high-dosage groups increased(P<0.05),loose stool rate,fecal water rate and AWR scores decreased(P<0.05),a small amount of neutrophils was visible in the colon tissue,the expressions of Occludin,ZO-1,Math-1 mRNA in colon tissue increased in Huosha Oral Liquid medium-and high-dosage groups(P<0.05)and the expressions of Notch1 and Hes-1 mRNA decreased(P<0.05),the expression of Notch1 protein decreased in Western medicine group and Huosha Oral Liquid groups(P<0.05).Conclusion Huosha Oral Liquid may regulate the expressions of tight junction proteins by inhibiting excessive activation of the Notch/Hes-1/Math-1 signaling pathway,thereby protecting the intestinal mucosal barrier function,improving diarrhea symptoms in IBS-D rats with cold and dampness syndrome,and reducing visceral sensitivity and colonic tissue inflammation.

Objective To observe the effects of Huosha Oral Liquid on irritable bowel syndrome with diarrhea(IBS-D)rats with cold and dampness syndrome based on Notch1/Hes-1/Math-1 signaling pathway;To elucidate its mechanism of repair of intestinal mucosal barrier.Methods An IBS-D cold and dampness syndrome rat model was established by using restraint stress combined with bitter-cold cathartic method and cold and dampness environmental method.After modeling,the rats were randomly divided into model group,Western medicine group and Huosha Oral Liquid low-,medium-and high-dosage groups,with 6 rats in each group.An additional 6 rats were set as the normal group.The Western medicine group was given mebeverine(13.5 mg/kg)by gavage,while the Huosha Oral Liquid low-,medium-and high-dosage groups were given Huosha Oral Liquid(3.15,6.3,12.6 g/kg)by gavage,the normal group and model group were given distilled water by gavage,for 4 weeks.The general status of rats were ovserved,the body mass,loose stool rate,fecal water rate and abdominal withdrawal reflex(AWR)score were measured,the morphalogy of colon tissue were observed by HE staining,the mRNA expression of Occludin,ZO-1,Notch1,Hes-1 and Math-1 in colon tissue were detected by RT-PCR,the protein expression of Notch1 was detected by Western blot.Results Compared with the normal group,the model group had a reduced body mass(P<0.05)and increased loose stool rate,fecal water rate and AWR score(P<0.05),inflammatory cell infiltration was observed in colon tissue and the expressions of Occludin,ZO-1,Math-1 mRNA decreased(P<0.05),the expressions of Notch1,Hes-1 mRNA increased(P<0.05),and the expression of Notch1 protein increased(P<0.05).Compared with the model group,the body mass of rats in Western medicine group and Huosha Oral Liquid low-,medium-and high-dosage groups increased(P<0.05),loose stool rate,fecal water rate and AWR scores decreased(P<0.05),a small amount of neutrophils was visible in the colon tissue,the expressions of Occludin,ZO-1,Math-1 mRNA in colon tissue increased in Huosha Oral Liquid medium-and high-dosage groups(P<0.05)and the expressions of Notch1 and Hes-1 mRNA decreased(P<0.05),the expression of Notch1 protein decreased in Western medicine group and Huosha Oral Liquid groups(P<0.05).Conclusion Huosha Oral Liquid may regulate the expressions of tight junction proteins by inhibiting excessive activation of the Notch/Hes-1/Math-1 signaling pathway,thereby protecting the intestinal mucosal barrier function,improving diarrhea symptoms in IBS-D rats with cold and dampness syndrome,and reducing visceral sensitivity and colonic tissue inflammation.

ObjectiveTo clarify the relationship between intestinal flora and intestinal motility in rats with slow transit constipation （STC） and qi stagnation syndrome by conducting a pseudo-sterile experiment and fecal microbiota transplantation （FMT） technology. MethodsTwenty-four Wistar rats were randomly divided into normal group （n=6）， STC with qi stagnation pattern group （n=6） and pseudo-sterile group （n=12）. In the STC group with qi stagnation pattern， 3 mg/kg of loperamide suspension by intragastric administration combined with tail clamping stimulation were performed to establish the rat model of STC with qi stagnation pattern. After successful modeling， fresh feces from the rats in the STC with qi stagnation pattern group and the normal group were collected to prepare 100 mg/ml of fecal bacterial suspension. In the pseudo-sterile group， the antibiotic cocktail method was used （a mixed antibiotic suspension containing bacitracin， streptomycin sulfate， and neomycin sulfate at 20 mg/ml each was administered intragastrically） to establish pseudo-sterile rats model. After successful modeling， the rats were randomly divided into normal fecal bacterial liquid group and STC with qi stagnation pattern fecal bacterial liquid group， with six rats in each group， and then were given 10 ml/kg of the prepared corresponding rat fecal bacterial suspension by gavage. Rats in STC with qi stagnation pattern group were given an equal volume of sterile water by gavage. All groups were administered once a day for 7 consecutive days. The small intestinal propulsion rate of the STC with qi stagnation pattern group， the normal fecal bacterial liquid group， and STC with qi stagnation pattern fecal bacterial liquid group were compared. ELISA method was used to detect serum 5-hydroxytryptamine （5-HT） levels. Immunohistochemistry was used to detect the positive expression levels of 5-hydroxytryptamine 3 receptor （5-HT3R） and 5-hydroxytryptamine 4 receptor （5-HT4R） in colon tissue. Western blot method was used to detect the protein expression levels of tryptophan hydroxylase 1 （TPH1）， tryptophan hydroxylase 2 （TPH2）， serotonin transporter （SERT）， and monoamine oxidase A （MAO-A） in colon tissue. ResultsCompared to those in the normal fecal bacterial liquid group， the small intestinal propulsion rate， serum 5-HT level， positive expression of 5-HT3R and 5-HT4R in colon tissue， and protein expression of TPH1， TPH2， SERT and MAO-A significantly decreased in the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＜0.05）. There was no statistically significant difference in the indicators between the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＞0.05）. ConclusionThe intestinal flora in STC rats with qi stagnation pattern can lead to a slowdown in intestinal transmission function， whose mechanism may be related to intestinal motility disorders affected by the synthesis， transport， metabolism and other pathways of 5-HT.

OBJECTIVE: To research the toxicokinetics features of ivabradine hydrochloride and the accumulation situation of its products in Beagles, during one month oral administration with it.