ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

This article is to illustrate the connection between neutrophil extracellular traps （NETs） and stasis-toxin interactions during the development of liver cancer， and to provide ideas for the prevention and treatment of liver cancer. It is believed that the mechanism of tumor-related thrombosis formation caused by NETs is closely related to "stasis" in traditional Chinese medicine （TCM）， and the continuous maintenance or enhancement of tumor microenvironment by NETs is closely related to "toxin" in TCM； the strong coagulation-promoting effect of NETs makes the local microenvironment， which can be regarded as the toxin caused by blood stasis； the changes of tumor microenvironment induced by NETs further induced platelet aggregation， which may present a state of stasis caused by toxin； the pathological changes of liver cancer caused by NETs can be regarded as the microscopic manifestation of the mechanism of stasis-toxin interaction of liver cancer.

Combining the knowledge of traditional Chinese medicine and modern medicine on glucolipid metabolism disorders， it is believed that the formation process of glycolipid metabolism disorders can be presented as five states "depression， phlegm-dampness， heat， blood stasis， and deficiency"， and the turbid pathogens run through the whole process. Accordingly， the method of "regulating states and removing turbidity" is proposed， which is specifically the method of resolving depression and turbidity， dispelling phlegm-dampness and turbidity， clearing heat and turbidity， dispelling blood stasis and turbidity， and replenishing deficiency and removing turbidity. Combined with the biological basis of glycolipid metabolism disorder， through the analysis of the clinical application of each method and the related mechanism of action， it is clarified that the method of regulating states and resolving turbidity can play a role in improving glycolipid metabolism disorder by regulating lipid metabolism disorder， insulin resistance， bile acid metabolism abnormality， intestinal bacterial flora， and its metabolite abnormality and other mechanisms of action.

OBJECTIVE To analyze the accumulation law of saponins during growth in Paris polyphylla Smith var. chinensis (Franch.) Hara, determine the content of the main saponins in different cultivation years, age groups, cultivation modes and provenances. METHODS The content of 5 kinds saponins(I, II, VI, VII, H) was simultaneous determined by HPLC. RESULTS The total saponins in P. polyphylla Smith var. chinensis (Franch.) Hara were mainly composed of saponin VII and H, supplemented by saponin VI, I and II. The content of saponins(I, II, VII) was significantly different among different cultivation years rhizome, while it reached the standard of Chinese Pharmacopoeia 2020 Edition after 6 years old, 8-year-old rhizome was the highest. The saponins(I, II, VII) content in 4a rhizome and 5a rhizome was significant higher than others, and it ranged from 0.354% to 0.765% in different cultivation modes, from high to low as follows: coniferous forest>bamboo forest>broadleaf forest>greenhouse. In different provenances, it ranged from 0.592% to 0.741%, reached the highest level in Qingyuan Baikan, and was slightly lower than the standard of Chinese Pharmacopoeia 2020 Edition in Sanming Fujian. CONCLUSION There are remarkable correlations among saponins accumulation amounts and cultivation years, age groups, cultivation modes and provenances, which can provide reference for the artificial culvication of P. polyphylla Smith var. chinensis (Franch.) Hara.

Objective To compare the pathological status of gastric mucosa and the expression of HH-PTCH-SMO-GLI(Hedgehog signaling pathway)and NOX/NF-κB/STAT1 signaling pathways in Hp and non-HP infected CAG patients,and to explore the biological mechanism of Hp promoting the"inflammatory cancer transformation"of CAG.Methods 43 patients with CAG who met the criteria were enrolled and divided into CAG with Hp infection group(Hp+ CAG group,n=21)and CAG without Hp infection group(HP-CAG group,n=22).The histological changes of gastric mucosa were observed by hematoxylin-eosin(HE)staining.Western blot was used to detect the relative expression levels of NOX1,NOX2,NOX4,STAT1,P65 and P-P65 in gastric mucosa.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect Gli1 mRNA,Gli2 mRNA,Gli3 mRNA,Shh mRNA,Smo mRNA,Ptch mRNA,NOX1 mRNA,NOX2 mRNA,NOX4 mRNA and NF-κB mRNA in gastric mucosa The mRNA level.Results HE staining results of gastric tissues in the two groups:In the Hp+CAG group,gastric epithelial cells were partially necrotic and shed,the surface was not smooth,the number of glands was reduced and disordered,intestinal metaplasia was observed,and diffuse lymphocyte and neutrophil infiltration were observed in the lamina proper.The degree of lymphocyte and neutrophil infiltration in HP-CAG group was lighter than that in Hp+CAG group.RT-qPCR results:Compared with HP-CAG group,the levels of Gli1 mRNA,Shh mRNA,Smo mRNA and Ptch mRNA in gastric mucosa of Hp+CAG group were significantly decreased(P<0.01).The levels of Gli2 mRNA,Gli3 mRNA,NOX1 mRNA,NOX2 mRNA,NOX4 mRNA and NF-κB mRNA were significantly increased(P<0.01).Western blot detection results:Compared with hP-CAG group,the relative expression levels of NOX1/GAPDH,NOX2/GAPDH,NOX4/GAPDH and P-P65/GAPDH in gastric mucosa of Hp+CAG group were significantly increased(P<0.01),and the STAT1 level was significantly decreased(P<0.01).There was no significant difference in the relative expression of P65/GAPDH between the two groups(P>0.05).Conclusion Hp infection may cause long-term inflammation of gastric mucosa,promote atrophy and intestinal metaplasia,and increase the risk of cancer by inhibiting hH-PTC-SMO-GLi signaling pathway and abnormal activation of NOX/NF-κB/STAT1 signaling pathway.

Nonalcoholic fatty liver disease （NAFLD） is an abnormal lipid metabolic disorder of the liver characterized by accumulation of a large amount of lipids in the liver， and it is currently the most common liver disease around the world. Mendelian randomization （MR） incorporates genomic data into traditional epidemiological study designs to infer the causal relationship between exposure factors and disease risk. In recent years， MR has been widely used in studies on inference of the etiology of NAFLD. This article systematically summarizes the advances in the application of MR in NAFLD research， so as to provide new ideas for understanding the nature of the disease and scientific interventions.

Objectives To explore the effect and mechanism of saikosaponin D on cellular autophagy of ICCs by regulating the CaMKKβ/AMPK signaling pathway.Methods Rat primary ICCs cells were isolated and stimulated with glutamate to construct an autophagy model.The Ca2+level was detected by immunofluorescence.Primary ICCs cells were divided into control group,model group,model+saikosaponin D group,model+CaMKKβ inhibitor group,and model+saikosaponin D+CaMKKβ inhibitor group.The ultrastructure of autophagosomes was observed by transmission electron microscopy.The levels of Ghrelin and SP were detected by ELISA.The expressions of Ca2+and LC-3Ⅱ were detected by immunofluorescence.The protein expression levels of LC-3Ⅱ/Ⅰ、CaMKKβ,p-AMPK,Drp1,MFN2,IP3R and RyR were detected by Western blot.Results The fluorescence expression of LC-3Ⅱ was enhanced.Saikosaponin D reduced the levels of CaMKKβ,AMPK and MFN2(P<0.01),and increased the levels of LC-3Ⅱ/Ⅰ、IP3R,RyR,Drp1,Ghrelin and SP(P<0.01).The effect of Saikosaponin D combined with CaMKKβ inhibitor STO-609 was more significant.Conclusion Saikosaponin D can mediate Ca2+outflow through the CaMKKβ/AMPK signaling pathway,and affect the expression of excessive autophagy and gastrointestinal motility-related factors in ICCs cells.

Objective:To discuss the medication, prescription law and academic thoughts of doctors in the Ming and Qing Dynasties for the treatment of abdominal tympanites based on the Yi An Lei Ju. Methods:The medical records of doctors in the Ming and Qing Dynasties for the treatment of tympanites contained in Yi An Lei Ju were retrieved, and the prescription information was extracted. Based on R4.2.1 and the TCM inheritance calculation platform V3.0, the frequency, property and taste, and meridian tropism, and efficacy categories of drugs were counted, and association rules analysis and clustering analysis were performed. Results:A total of 214 prescriptions were included, involving 239 kinds of Chinese materia medica, with a total frequency of 2 080 times. The high-frequency drugs (frequency≥30) were Poria, Citri Reticulatae Pericarpium, Alismatis Rhizoma, Atractylodis Macrocephalae Rhizoma, Aconiti Lateralis Radix Praeparata, etc. The medicinal properties of the drugs were mainly warm, neutral and cold. The flavors were mainly pungent, sweet and bitter. The meridians were mainly spleen, liver and kidney meridians. The efficacy categories were mainly tonic drugs, diuresis and dampness drugs, and qi-regulating drugs. Association rule analysis found that the most frequently used drug pair was Poria-Citri Reticulatae Pericarpium. Clustering analysis showed that drugs (frequency≥20) could be divided into 4 clusters.Conclusions:Doctors in the Ming and Qing Dynasties treated abdominal tympanites with the basic treatment methods of invigorating the spleen and promoting diuresis, soothing the liver and regulating qi, warming the kidney and resolving fluid retention. The medication took into account the ascending and descending of qi, the combination of attack and supplement, the combination of cold and heat, and the harmony of qi and blood.

In recent years， the continuous advances in material sciences and techniques have helped with the establishment and development of liver organoids that can simulate the structure and function of organs in vivo. In addition to the research on traditional biological factors， the construction of microenvironments with different mechanical cues to investigate the influence of mechanical stimulation on the growth of liver organoids has also become a research focus. This article first discusses the development of liver organoids and then reviews the influence of mechanical forces of different properties on the formation of liver organoids， so as to lay a foundation for the construction of more complex and ordered liver organoids in vitro and provide ideal research models for understanding the interaction between biological and mechanical factors in the formation of liver organoids.