Objective:To observe the therapeutic effects of Danggui Shaoyao Powder on diarrhea type irritable bowel syndrome (IBS-D) rats with liver stagnation and spleen deficiency syndrome, as well as its effects on farnesyl ester X receptor (FXR) and G protein bile acid coupled receptor 5 (TGR5).Methods:Totally 60 SD rats were divided into normal group, model group, Deshute group, and Danggui Shaoyao Powder high-, medium-, and low-dosage groups using a random number table method, with 10 rats in each group. Except for the normal group, the IBS-D model of liver depression and spleen deficiency syndrome was established in all other groups using the method of mother child separation + chronic restraint stress + senna leaf gavage. After the completion of modeling, the Deshute group was treated with 1.5 g/kg for gavage. Danggui Shaoyao Powder high-, medium-, and low-dosage groups were orally administered with Danggui Shaoyao Powder at dosages of 1.93, 3.85, and 7.70 g/kg, respectively, while the normal group and model group were orally administered with equal volumes of physiological saline for gavage. The number of fecal particles and loose stools in each group of rats was observed, the loose stools rate was calculated, the small intestine carbon powder propulsion rate was measured, and the visceral sensitivity of rats was evaluated through abdominal wall withdrawal reaction (AWR) score. Open field experiments and sugar preference experiments were used to observe changes in depression and anxiety like behavior in rats. The level of D-xylose in serum was measured using phloroglucinol method, and the total bile acid (TBA) content in fecal supernatant was measured using ELISA method. Western-blot and RT-PCR were used to detect the protein and mRNA expressions of FXR and TGR5 in colon tissue.Results:Compared with the model group, Danggui Shaoyao Powder high-, medium-, and low-dosage groups and the Deshute group showed a decrease in loose stools rate, small intestine carbon powder propulsion rate and AWR score ( P<0.05), an increase in the number of passages and standing times of the open field experiment ( P<0.05), an increase in sugar preference rate ( P<0.05), an increase in serum D-xylose levels, a decrease in fecal TBA level ( P<0.05), and an increase in protein and mRNA expressions of FXR and TGR5 in colon tissue ( P<0.05). Conclusions:Danggui Shaoyao Powder can regulate intestinal motility disorders, reduce visceral sensitivity, and improve depression and anxiety in IBS-D rats with liver depression and spleen deficiency syndrome. Its mechanism may be related to activating the FXR/TGR5 signaling pathway and regulating bile acid metabolism in the body.

Objective:To establish the cisplatin-resistant human lung adenocarcinoma cell line A549/(DDP) cisplatin and to study the relationship between EHD2 and drug resistance. Methods:DDP-resistant human lung cancer cell line A549/DDP was established by gradual and stepwise dose enhancement. MTT was used to measure drug sensitivity. Western blot and immunofluorescence were used to evaluate expression and subcellular localization of EHD2 and breast cancer resistance protein (BCRP). Results:The DDP-resistant cell line A549/DDP was established, with a resistance index of 7.6. EHD2 and BCRP expressions both increased and were enriched on the cell surface membrane. Conclusion:Both EHD2 and BCRP expressions were enriched on the resistant cell surface membrane, suggesting that EHD2 endocytic protein stabilizes BCRP and is involved in drug resistance.

BACKGROUNDTo investigate the origin of tumor blood vessel and blood supply during pulmonary carcinogenesis, and the relationship between vascular endothelial growth factor (VEGF), its receptor Flk-1 and angiogenesis.

METHODSOne hundred Wistar rats were instilled with 3-methylcholanthrene (MCA) and diethylinitrosamine (DEN) to induce pulmonary squamous cell carcinoma through left lower lobe bronchus. To acquire different pathological phase during the carcinogenesis, rats were killed in 15, 35, 55, 65, 75 days after instillation. Yellow and green silastics were respectively injected into the bronchial and pulmonary arteries of 30 rats in 55, 65, 75 days after instillation. Intertumor microvessel density (MVD) was marked by anti-von Willebrand factor monoantibody. VEGF and Flk-1 expression were examined by immunohistochemistry.

RESULTSIn the tumor area the tumor blood vessels were yellow and connected with distorted bronchial artery and very few green incomplete branches of pulmonary artery were seen. Silastic particles could be seen in the disordered tumor blood vessels by microscope after bronchial artery perfusion. There was no silastic particles in the carcinoma interstitial blood vessels after pulmonary artery perfusion. MVD count significantly increased in carcinoma in situ (39.50±12.60) and infiltrative carcinoma (61.05±19.92) as compared to atypical hyperplasia (8.92±3.80)(both P < 0.01), and the increased vessels originated from bronchial artery, but not pulmonary artery. The expression of VEGF and Flk-1 increased during pulmonary carcinogenesis. The positive coefficients of VEGF and FLK-1 expressions became higher and higher from epithelial proliferation to squamous metaplasia, to atypical hyperplasia, to carcinoma in situ and finally to infiltrative carcinoma. There was significant correlation between MVD and VEGF expression (r=0.979 8, P < 0.005), as well as between MVD and Flk-1 expression (r=0.907 8, P < 0.05).

CONCLUSIONSAngiogenesis is the important phenomenon of the rat pulmonary carcinogenesis and the newly formed blood vessels in tumor connect with the branches of bronchial artery, but not pulmonary artery. This confirms that the blood supply of pulmonary carcinoma is from bronchial artery, not from pulmonary artery. VEGF and Flk-1 are closely related to angiogenesis of tumor.

Objective　To evaluate significance of squamous cell carcinoma associated antigen (SCC Ag) and carcinoembrynoic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3) in early diagnosis of lung squamous cell carcinoma. Methods　Lung squamous cell carcinoma was induced with methylcholanthrene (MCA) in iodized oil in Wistar rats. Ninty-one rats were used for this experiment, and the rats were sacrificed on the 20th, 40th, 50th, 60th, 70th and 80th days respectively. The stage of the cancer development including atypical hyperplasia, carcinoma in situ, early invasive carcinoma was diagnosed histopathologically. SCC Ag, CEA and CA15-3 in the serum of different conceration stage rats were measured by MEIA technique. Results　Serum levels of SCC Ag, CEA and CA15-3 of the model rats in the atypical hyperplasia and carcinoma in situ had no significant difference. The level of SCC Ag increased predominantly at the early invasive carcinoma(P＜0.01), but the other two markers had no change. Conclusion　SCC Ag level in serum may be of considerable importance in the early diagnosis of lung squamous cell carcinoma.