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ObjectiveMolecular docking and animal experiments were employed to explore the protective effect and mechanism of Da Chengqitang （DCQD） on intestinal barrier in septic mice. MethodText mining method was used to screen the active ingredients in DCQD. AutoDock Tools and Discovery Studio were used to study the interactions of active components with the core target proteins ［claudin-1， tumor necrosis factor （TNF）-α， interleukin （IL）-6， endogenous antimicrobial peptide mCRAMP， Toll-like receptor 4 （TLR4）， and myeloid differentiation primary response gene 88 （MyD88）］ in sepsis. Fifty C57BL/6 mice were randomized into sham， model， low- and high-dose （4 g∙kg-1 and 8 g∙kg-1） DCQD， and ulinastatin groups （n=10）. Before， during， and after the day of modeling surgery， each group was administrated with corresponding drugs. The mice in other groups except the model group were subjected to modeling by cecal ligation and puncture. Enzyme-linked immunosorbent assay （ELISA） was used measure the serum level of D-lactic acid to assess intestinal mucosa permeability. Hematoxylin-eosin staining was employed to observe the histopathological changes in the ileum and assess the intestinal mucosal damage and inflammatory infiltration. Western blotting was employed to determine the expression levels of tight junction proteins claudin-1 and occludin in the ileal tissue， which were indicative of the bowel barrier function. The TNF-α and IL-6 levels were measured by ELISA to assess the intestinal inflammation. The expression of mCRAMP in the ileal tissue was observed by immunohistochemistry. The mRNA levels of mCRAMP， TLR4， and MyD88 in mouse ileal tissue were determined by Real-time polymerase chain reaction， on the basis of which the mechanism of DCQD in protecting the intestinal barrier of septic mice was explored. ResultMolecular docking results showed that most of the 10 active ingredients of DCQD that were screened out by text mining could bind to sepsis targets by van der Waals force， hydrogen bonding， and other conjugated systems. The results of animal experiments showed that compared with the model group， low- or high-dose DCQD lowered the D-lactic acid level in the serum （P<0.01）， alleviated damage to the ileal tissue and mucosal edema， protected the small intestine villus integrity， reduced inflammatory cell infiltration， promoted the expression of claudin-1 （P<0.01）， lowered the IL-6 level （P<0.01）， up-regulated the mRNA and protein levels of mCRAMP （P<0.01）， and down-regulated the mRNA and protein levels of TLR4 and MyD88 （P<0.01） in the ileal tissue. In addition， high-dose DCQD lowered the TNF-α level and promoted the expression of occludin in the ileum tissue （P<0.01）， and low-dose DCQD up-regulated the protein level of occludin in the ileum tissue （P<0.05）. ConclusionDCQD has a protective effect on intestinal barrier in septic mice. It can reduce intestinal inflammation， repair intestinal mucosal damage， improve the tight junction protein level， and reduce intestinal mucosal permeability by up-regulating the mRNA and protein levels of mCRAMP and the down-regulating the expression of genes in the TLR4/MyD88 pathway.
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OBJECTIVE To study the protective effects of Dachengqi decoction （DCQD） on intestinal septic mice， and to explore the possible mechanisms from the Toll-like receptor 4（TLR4）/myeloid differentiation factor 88（MyD88） signaling pathway. METHODS The SPF male C57BL/6J mice were randomly divided into Sham group， Sham+DCQD-H group， model （CLP） group， DCQD-L group， DCQD-H group and Positive group. The model of intestinal sepsis was established by cecal ligation and puncture in CLP group， DCQD-L group， DCQD-H group and Positive group. Three days before the operation and seven days after the operation， DCQD-L group and DCQD-H group were given DCQD intragastrically at 4， 8 g/kg （calculated by crude drug）， respectively. Positive group was given ulinastatin intraperitoneally 2 h before operation and 7 d after the operation （at 50 000 U/kg）. In Sham group and Sham+DCQD-H group， only cecum of mice was exposed without ligation and puncture. Sham+DCQD- H group was given DCQD intragastrically （8 g/kg，calculated by crude drug） 3 days before the operation and 7 days after the operation. Both the Sham group and CLP group were given normal saline 0.2 mL intragstrically and intraperitoneally each day， for 10 consecutive days. After the operation， the severity of sepsis was assessed， and the 7 d survival rate of mice was assessed. One hour after the last medication， the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum and ileum of mice were determined； the pathological and morphological changes of mice’s liver， lung， kidney and ileum were observed； mRNA expressions of the TLR4 and MyD88 in ileum were tested. RESULTS Compared with CLP group， sepsis score， the levels of TNF-α and IL-6 in serum and ileum （except for IL-6 in ileum of DCQD-L group）， damage score of the liver， lung， kidney and ileum， mRNA expressions of TLR4 and MyD88 in ileum were all decreased significantly in DCQD-L group and DCQD-H group （P＜0.05 or P＜0.01）， while 7 d survival rate （except for DCQD-L group） was increased significantly （P＜0.05）. The damage to liver tissue in mice was significantly improved， and inflammation infiltration and apoptosis were reduced； lung tissue damage had been alleviated， with varying degrees of improvement in alveolar atrophy， bleeding and edema； the renal tissue damage was improved and weakened dilation of renal tubular lumen was weakened； the damage and edema of ileal tissue were significantly improved. CONCLUSIONS DCQD may exert a protective role on intestinal septic model mice. The mechanism may be related to the inhibition of systemic inflammation， the reduction of multiple organ damage， and down-regulation of TLR4/MyD88 signaling pathway.
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Objective:To study the influencing factors in predicting conversion of laparoscopic liver tumor resection to open surgery to provide references for selection of patients for laparoscopic hepatectomy.Methods:The clinical data of 222 patients who were planned to undergo laparoscopic liver tumor resection at Ningbo Medical Center Lihuili Hospital from December 2015 to November 2018 were analyzed retrospectively. There were 146 males and 76 females, with an average age of 58.1 years. These patients were divided into the conversion group ( n=24) and the laparscopic group ( n=198) according to whether intraoperative conversion to open surgery was carried out. Chi-square test was used to evaluate relevant factors of conversion. Logistic regression analysis was used for multivariate analysis and to find out independent risk factors of conversion. Results:The conversion rate was 10.8% (24/222). Univariate analysis showed that hypertension, history of abdominal surgery, liver cirrhosis, tumors located at posterosuperior segments and major liver resection were significantly associated with conversion (all P<0.05). Multivariate analysis showed that the risk factors which were independently associated with conversion were liver cirrhosis ( OR=2.611, 95% CI: 1.018-6.701), tumors located at posterosuperior segments ( OR=6.115, 95% CI: 2.207-16.941), and major liver resection ( OR=3.361, 95% CI: 1.150-9.825)(all P<0.05). Conclusion:Patients who were planned for laparoscopic liver tumor resection with liver cirrhosis, tumors at posterosuperior segments or major liver resection were at higher risks of conversion to open surgery.
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Objective To establish a method of HPLC assay for determining stilbene glucoside in Zishen Ningshen Pills(ZNP),and to study the influence factors on the content of stilbene glucoside in the process of preparation.Methods HPLC was used for the determination of stilbene glucoside in ZNP.Through simulation the process of preparation,the stilbene glucoside content in the intermediate products was determined by HPLC,and its retention rate and metastasis rate were also investigated.Results The resolution and the linearity of stilbene glucoside were fine,the average recoveries being 98 % ～ 102 %.The retention rate of stilbene glucoside in the drying powder was 60.3 %,lower than that in the original medicinal powder.Conclusion The quantitative method for determining the ingredients in ZNP is simple,feasible and reproducible,and is beneficial for quality control of ZNP.The drying process under normal pressure is the main influence factors of the decrease of stilbene glucoside content,and the decompression drying can be taken into account to take the place of the atmospheric drying.
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Objective To establish a HPLC method for the determination of effective components in Radix Notoginseng in San Xiong Zhitong Ointment.Methods Kromasil KR100-5 C18 column(250 mm?4.6 mm,5 ?m)and gradient elution were used.Solvent A was acetonitrile and solvent B was water.The detection wavelength was set at 203 nm.Results The four components were isolated well.The linearity was fine with the recoveries of 98 %～100 %.Conclusion The quantitative method for determining the ingredient of San Xiong Zhitong Ointment is simple,feasible and repeatable,and is beneficial for quality control of San Xiong Zhitong Ointment.