ObjectiveTo explore the mechanism of Yinchenhao Tang regulating the tumor necrosis factor receptor superfamily member 6 （Fas）/cysteine protease-8 （Caspase-8）/cysteine protease-3 （Caspase-3） signaling pathway to inhibit hepatocyte apoptosis and improve cholestatic liver injury （CLI）. MethodsAmong 48 Wistar rats，12 rats were randomly selected as the blank group，and the other rats were administered alpha-naphthalene isothiocyanate （ANIT） by gavage to induce a CLI model. The modeling rats were randomly divided into the model group， the ursodeoxycholic acid group（0.1 g·kg^-1） and the Yinchenhao Tang group（9.23 g·kg^-1），with 12 rats in each group. The rats in each group were given corresponding drugs by gavage for three consecutive days. The levels of alanine aminotransferase （ALT），aspartate aminotransferase （AST），alkaline phosphatase （ALP），gamma-glutamyl transpeptidase （γ-GT），total bilirubin （TBil） and total bile acid （TBA） in serum were detected. The levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in liver tissue were detected. The histopathological changes of the liver were observed by hematoxylin-eosin （HE） staining. The protein and mRNA expressions of Fas，Caspase-8，Caspase-3，B-cell lymphoma-2 （Bcl-2） associated X protein （Bax） and Bcl-2 in liver tissue were detected by Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultsCompared with those in the blank group，the levels of ALT，AST，ALP，γ-GT，TBA and TBil in serum of the model group were significantly increased （P<0.01）. The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly increased （P<0.01）. The arrangement of hepatocytes was disordered，and inflammatory cell infiltration and bile duct epithelial cell proliferation were observed. The protein and mRNA expressions of Fas，Caspase-8，Caspase-3 and Bax in liver tissue were significantly increased（P<0.05，P<0.01），while the protein and mRNA expressions of Bcl-2 were significantly decreased （P<0.05，P<0.01）. Compared with those in the model group，the levels of ALP，γ-GT，TBA and TBil in the serum of rats in the ursodeoxycholic acid group were significantly decreased. The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly decreased（P<0.05，P<0.01）. The protein and mRNA expressions of Fas，Caspase-8，Caspase-3 and Bax in liver tissue were significantly decreased （P<0.05，P<0.01），while the mRNA expression of Bcl-2 was significantly increased （P<0.05，）. The levels of ALT，AST，γ-GT，TBA and TBil in the serum of rats in the Yinchenhao Tang group were significantly decreased （P<0.01）. The levels and mRNA expressions of TNF-α and IL-1β in liver tissue were significantly decreased （P<0.05，P<0.01）. The protein expression of Fas and Bax and the mRNA expression of Fas，Caspase-8，Caspase-3 and Bax in liver tissue were significantly decreased （P<0.05，P<0.01），while the protein and mRNA expression of Bcl-2 were significantly increased （P<0.05，P<0.01）. Hepatocyte injury，inflammatory cell infiltration and proliferation of bile duct epithelial cells were reduced. ConclusionYinchenhao Tang can ameliorate CLI，and its mechanism may be related to inhibiting hepatocyte apoptosis mediated by the Fas/Caspase-8/Caspase-3 signaling pathway.

ObjectiveTo study the mechanism of Yinchenhao Tang on the improvement of cholestatic liver injury （CLI） by inhibiting toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor-κB （NF-κB） pathway via regulating farnesol X receptor （FXR）. MethodsA total of 40 Wistar male rats were randomly selected， with 10 as a blank group，and the remaining rats were subjected to the CLI model induced by alpha-naphthalene isothiocyanate （ANIT）. After modeling，they were randomly divided into the model group， the ursodeoxycholic acid （0.1 g·kg^-1） group and the Yinchenhao Tang （9.23 g·kg^-1） group，with 10 animals in each group. Each administration group was given the corresponding drug by intragastric administration for three consecutive days. Alanine aminotransferase （ALT），aspartate aminotransferase （AST），alkaline phosphatase （ALP），γ-glutamyl transpeptidase （γ-GT），total bile acid （TBA），total bilirubin （TBil） and direct bilirubin （DBil） levels in serum were detected. Tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β），and interleukin-6 （IL-6） levels in liver tissue were detected. Real-time PCR was used to detect the mRNA expression of FXR，TLR4，MyD88，NF-κB，F4/80，TNF-α，IL-1β and IL-6 in liver tissue. Western blot was used to detect protein expression of FXR，TLR4，MyD88 and NF-κB in liver tissue. The histopathological changes of the liver were observed by hematoxylin-eosin （HE） staining. ResultsCompared with those in the blank group，ALT，AST，ALP，γ-GT，TBA，TBil and DBil levels in serum of rats in the model group were significantly increased （P<0.01）. The levels and mRNA expression of TNF-α，IL-1β and IL-6 in liver tissue were significantly increased （P<0.01），and the mRNA and protein expressions of FXR in liver tissue were decreased （P<0.01）. The mRNA and protein expressions of TLR4，MyD88 and NF-κB and the mRNA expression of F4/80 were obviously increased （P<0.05，P<0.01）. Hepatic histopathology showed inflammatory cell infiltration and proliferative changes of bile duct epithelial cells. Compared with those in the model group，ALT，ALP，γ-GT，TBA，TBil and DBil levels in serum of rats in the ursodeoxycholic acid group were obviously decreased （P<0.05，P<0.01），and the levels and mRNA expression of TNF-α，IL-1β and IL-6 in liver tissue were obviously decreased （P<0.05，P<0.01）. The mRNA and protein expressions of TLR4，MyD88 and NF-κB and the mRNA expression of F4/80 in liver tissue were obviously decreased （P<0.05，P<0.01）. ALT，AST，ALP，γ-GT，TBA，TBil and DBil levels in the serum of rats in the Yinchenhao Tang group were obviously decreased （P<0.05，P<0.01），and the levels and mRNA expression of TNF-α，IL-1β and IL-6 in liver tissue were obviously decreased （P<0.01）. The mRNA and protein expressions of FXR in liver tissue were significantly increased，and the mRNA expressions of TLR4，MyD88，NF-κB，and F4/80， as well as the protein expressions of TLR4 and NF-κB were obviously decreased （P<0.05，P<0.01）. The inflammatory cell infiltration of liver tissue and the proliferation of bile duct epithelial cells decreased. ConclusionYinchenhao Tang has an obvious protective effect on CLI，and its mechanism may be related to regulating FXR to inhibit TLR4/MyD88/NF-κB pathway-mediated inflammatory response.

OBJECTIVES: To study the therapeutic mechanism of Tuihuang Mixture against cholestasis. METHODS: Forty-eight Wistar rats were randomized equally into blank group, model group, ursodeoxycholic acid group and Tuihuang Mixture group. Except for those in the blank group, all the rats were given α‑naphthylisothiocyanate (ANIT) to establish rat models of cholestasis, followed by treatments with indicated drugs or distilled water. Serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL of the rats were determined, and hepatic expressions IL-1β, IL-18, FXR, NLRP3, ASC, Caspase-1 and GSDMD were detected using q-PCR, ELISA or Western blotting. Histopathological changes of the liver tissues were observed using HE staining. RESULTS: The rat models of cholestasis had significantly increased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL with increased mRNA and protein expressions of IL-1β and IL-18, decreased protein and mRNA expressions of FXR, and increased protein expressions of NLRP3 and Caspase-1 and mRNA expressions of NLRP3, ASC, Caspase-1 and GSDMD in the liver tissue, showing also irregular arrangement of liver cells, proliferation of bile duct epithelial cells and inflammatory cells infiltration. Treatment of the rat models with Tuihuang Mixture significantly decreased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL, lowered IL-1β and IL-18 and increased FXR protein and mRNA expressions, and reduced NLRP3, ASC, Caspase-1 and GSDMD proteins and NLRP3, ASC and Caspase-1 mRNA expressions in the liver tissue. Tuihuang Mixture also significantly alleviated hepatocyte injury, bile duct epithelial cell proliferation and inflammatory cell infiltration in the liver of the rat models. CONCLUSIONS Tuihuang Mixture can effectively improve cholestasis in rats possibly by inhibiting NLRP3 inflammatosome-mediated pyroptosis via regulating FXR.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein ; Rats ; Receptors, Cytoplasmic and Nuclear/metabolism* ; Cholestasis/drug therapy* ; Rats, Wistar ; Inflammasomes/metabolism* ; 1-Naphthylisothiocyanate ; Drugs, Chinese Herbal/therapeutic use* ; Male ; Interleukin-18/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

Abstract Cholestasis can be seen in many acute and chronic liver diseases. If not intervened in time, persistent cholestasis can further progress to liver fibrosis, liver cirrhosis, liver failure, and even death. The pathogenesis of cholestasis is complex, and there is currently a lack of effective therapeutic drugs. Bile acids(BAs), the main component of bile, are synthesized from cholesterol in the liver as primary bile acids. After entering the intestine through the enterohepatic circulation, they are reshaped into secondary bile acids by gut microbiota. BAs can directly or indirectly affect the composition and function of gut microbiota, which in turn can regulate the synthesis and metabolism of BAs. The interaction between BAs and gut microbiota plays an important role in the pathogenesis and treatment of cholestasis. This review elucidates the bidirectional regulatory mechanisms between BAs and gut microbiota and their roles in the pathogenesis and treatment of cholestasis, aiming to provide insights and references for basic research and clinical practice related to cholestasis-associated diseases.

Objective: To compare the clinical effects of conventional hysteroscopy and HEOS hysteroscopy in the treatment of refractory intrauterine residuals. Methods: From January 2017 to February 2019, 60 patients with refractory intrauterine pregnancy residuals admitted to the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were randomly selected.The patients were divided into traditional endoscopy group and HEOS system endoscope group according to the different operation method, with 30 cases in each group.The operation time, intraoperative blood loss, hospitalization cost, postoperative uterine adhesion rate, postoperative menstrual recovery time, postoperative recovery rate of endometrium at the first, second and third month after operation were compared between the two groups. Results: The operation time of the conventional laparoscopic group was (24.38±3.16)min, which was longer than that of the HEOS system hysteroscopy group[(18.71±3.32)min](t=1.336, P=0.027). The intraoperative blood loss of the conventional laparoscopic group was (18.41±4.35)mL, which was higher than that of the HEOS system hysteroscopy group[(12.81±3.84)mL](t=1.853, P=0.031). The hospitalization cost of the conventional laparoscopic group was (7 461.87±344.92)CNY, which was more than that of the HEOS system hysteroscopy group[(6 743.19±298.52)CNY](t=0.037, P=0.040). In addition, the uterine adhesion rate in the traditional hysteroscopic group was 20.00%(6/30), which was higher than that in the HEOS system hysteroscopy group[6.67%(2/30)](χ²=2.308, P=0.029), and the postoperative menstrual recovery time in the traditional hysteroscopic group was (31.46±4.12)d, which was longer than that in the HEOS system hysteroscopy group[(26.18±3.72)d](t=1.730, P=0.026). At the first, second and third month after operation, the good endometrial recovery rates in the traditional laparoscopic group were 20.00%(6/30), 43.33%(13/30), 63.33%(19/30), respectively, which were lower than those in the HEOS system hysteroscopy group[46.67%(14/30), 70.00%(21/30), 93.33%(28/30)](χ²=4.800, 4.344, 7.954, P=0.028, 0.037, 0.005). Conclusion HEOS system endoscope in the treatment of refractory intrauterine residual has good effect and is worthy of promoting.

Objective: To investigate the expression of integrin β₃ in the endometrial implantation window of patients with cesarean scar and infertility, and its correlation with endometrial receptivity. Methods: A total of 40 patients with previous cesarean scar defect (PCSD) secondary infertility treated in Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from April 2018 to December 2019 were enrolled as the observation group.The natural cycle of 40 normal women who were examined in our hospital at the same time were selected as the control group.The two groups were enrolled in the endometrium of the glandular epithelial cells for MMP-9, TIMP-1 and LIF immunohistochemical staining, and statistical analysis was performed.The serum E₂ and P levels of the implantation group were compared.The E₂/P ratio was measured and compared, and the expression level of integrin β₃ endometrial planting window was compared. Results: The MMP-9, TIMP-1 and LIF in the observation group [(175.31±56.36), (201.46±51.34), (209.23±45.23)] were significantly lower than those in the control group [(252.35±78.43), (257.23±74.13), (298.34±72.35)] (t=5.334, 5.766, 6.023, all P<0.05), but the E₂, P and E₂/P in the observation group [(515.31±56.36)pmol/L, (53.71±8.34)pmol/L, (13.23±5.23)] were higher than those in the control group[(352.35±78.43)pmol/L, (33.13±4.13)pmol/L, (8.17±2.91)] (t=7.334, 4.251, 3.241, all P<0.05). The level of beta 3 in the observation group (0.163±0.013) was significantly lower than that in the control group (0.253±0.031) (t=4.342, P<0.05). The thickness of endometrium in the observation group [(9.12±2.03)mm] was significantly thinner than that in the control group [(12.24±2.45)mm] (t=3.226, P<0.05). Conclusion The expression of integrin β₃ in the endometrial implantation window of patients with cesarean section scar infertility has a certain influence on endometrial receptivity.

Objective: To explore the diagnostic value of serum NSE, S100B protein and myocardial zymogram in premature infants with intrauterine infection. Methods: From January 2016 to December 2017, 60 preterm infants with intrauterine infection in the Integrated Chinese and Western Medicine Hospital of Wenzhou were selected in the study.According to whether brain injury occurred, they were divided into brain injury group (28 cases) and non-brain injury group (32 cases). Serum NSE content was detected by chemiluminescence method, serum S100B protein level was detected by enzyme linked immunosorbent assay (ELISA), and serum CK and HBDH levels were detected by automatic biochemical analyzer.The serum levels of NSE, S100B, CK and HBDH were compared between the two groups, the combined diagnostic efficacy of NSE+ S100B protein+ CK+ HBDH was analyzed, the correlation of serum NSE, S100B protein, CK, HBDH with brain injury wasanalyzed. Results: The levels of serum NSE [(2.43±0.54)μg/L] and S 100B [(14.36±3.21)ng/L] in the brain injury group were higher than those in the non-brain injury group [(0.97±0.27)μg/L and (8.10±1.87)ng/L] (t=13.498, 9.370, all P<0.05). The levels of serum CK [(437.64±54.12)U/L] and HBDH [(387.91±56.45)U/L] in the brain injury group were significantly higher than those in the non-brain injury group [(183.54±32.58)U/L and (174.3±26.63)U/L] (t=22.347, 19.126, all P<0.05). The sensitivity and specificity of the combined diagnosis of NSE+ S100B protein and myocardial zymogram were higher than those of each single index.Serum NSE, S100B protein, CK and HBDH were positively correlated with brain injury. Conclusion The elevation of serum NSE, S100B protein and myocardial zymogram in preterm infants with intrauterine infection after birth has certain clinical significance in judging whether brain injury occurs or not.

Objective To investigate the change of local vaginal microenvironment in atrophic vaginitis after estrogen therapy.Methods A prospective study was conducted in 128 patients with atrophic vaginitis.The patients were treated with estriol cream by vaginal administration,and the efficacy was observed after treatment.Results The PH value was decreased and the detection rate of lactobacilli was significantly increased after estrogen therapy(P＜0.01).Average vaginal health score was also increased (P＜0.01).Interleukin-6 (IL 6) level in vaginal secretion was decreased after the treatment (P＜0.01).Interferon-γ (INF-γ) level was increased obviously after treatment,while there was no significant difference between the two groups(P＞0.05).Conclusions Estrogen can significantly improve the local vaginal microenvironment in atrophic vaginitis.Estrogen may regulate the immune function of vagina by reversing the imbalance of Th1/Th2.

Objective To investigate the effect of cilazapril on reactivit y of blood vessels in diabetic rats. 　Methods The mesenteric vessels of streptozotocin induced diabetic rats wer e perfused with Kreb-Ringer's solution in vitro,the mesenteric vessels reactivi ty of diabetic rats was measured by physiological recorder. 　Results The responsive continual time to norepinephrine was delay ed signific antly(9.4±3.1 vs 5.2±2.1,P<0.01) and the highest perfusion pressu re decrease significantly in diabetic rats(5.12±0.87 vs 7.81±0.92 kPa, P<0.01); When taken cilazapril showed no difference in these indexes in diabetic animal s.