ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

ObjectiveTo investigate the mechanism by which Xianglian Huazhuo prescription alleviates chronic atrophic gastritis （CAG） through modulating the nuclear factor erythroid 2-related factor 2 （Nrf2）/quinone oxidoreductase 1 （NQO1）/heme oxygenase-1 （HO-1） pathway of oxidative stress （OS） based on animal experiments. MethodsFifty rats were modeled for CAG by gavage of 2% sodium salicylate （10 mL·kg^-1）， free access to 180 mg·L^-1 N-methyl-N′-nitro-N-nitrosoguanidine （MNNG）， and periodic diet intervention （72 h cycle： 24 h fasting + 48 h free access to food）. The modeled rats were grouped by a random number method as follows： low-， medium-， and high-dose （9， 18，36 g·kg^-1， respectively） Xianglian Huazhuo prescription， Moradan， and model， with 10 rats in each group. The general state of all rats was observed. Hematoxylin-eosin （HE） staining was employed to observe the pathological changes of the gastric mucosal tissue. The ultrastructure of cells was observed under transmission electron microscopy. Biochemical methods were adopted to determine the levels of malondialdehyde （MDA） and total superoxide dismutase （T-SOD） in the rat serum. The mRNA and protein levels of Kelch-like ECH-associated protein 1 （Keap1）， Nrf2， NQO1， and HO-1 in the gastric mucosal tissue of rats from each group were measured by Real-time polymerase chain reaction （Real-time PCR） and Western blot， respectively. Reactive oxygen species （ROS） staining was employed to detect the fluorescence intensity of ROS in the gastric mucosal tissue of rats in each group. ResultsCompared with the normal group， the modeled rats were sluggish， had yellowing and sparse hair， short and dark tail， loose stool， and reduced food intake， and showed the gastric mucosa characterized by atrophy， thinning， congestive edema， declined gland density and disarrangement， and apparent inflammatory cell infiltration， irregular nucleus morphology， nuclear condensation， mitochondria with decreased number， declined density， and abnormal morphology， vacuolization， declined serum T-SOD level （P<0.05）， elevated serum MDA level （P<0.05）， downregulated mRNA and protein levels of Nrf2， NQO1， and HO-1 （P<0.05）， upregulated mRNA and protein levels of Keap1 （P<0.05）， and enhanced mean fluorescence intensity of ROS （P<0.05）. Compared with the model group， each treatment group exhibited improved general state， alleviated histopathological changes in gastric mucosa， improved ultrastructural characteristics of cells， elevated T-SOD level and declined MDA level in the serum （P<0.05）， upregulated mRNA and protein levels of Nrf2， NQO1， and HO-1， downregulated mRNA and protein levels of Keap1 （P<0.05）， and attenuated mean fluorescence intensity of ROS （P<0.05）. ConclusionXianglian Huazhuo prescription can improve the general status， ameliorate pathological changes of gastric mucosa， and repair the cell ultrastructure to varying degrees， demonstrating a definite therapeutic effect on CAG. It may restrain OS and safeguard the natural physiological function of gastric mucosa in CAG rats by regulating the Nrf2/NQO1/HO-1 signaling pathway.

ObjectiveTo investigate the mechanism by which Xianglian Huazhuo prescription alleviates chronic atrophic gastritis （CAG） through modulating the nuclear factor erythroid 2-related factor 2 （Nrf2）/quinone oxidoreductase 1 （NQO1）/heme oxygenase-1 （HO-1） pathway of oxidative stress （OS） based on animal experiments. MethodsFifty rats were modeled for CAG by gavage of 2% sodium salicylate （10 mL·kg^-1）， free access to 180 mg·L^-1 N-methyl-N′-nitro-N-nitrosoguanidine （MNNG）， and periodic diet intervention （72 h cycle： 24 h fasting + 48 h free access to food）. The modeled rats were grouped by a random number method as follows： low-， medium-， and high-dose （9， 18，36 g·kg^-1， respectively） Xianglian Huazhuo prescription， Moradan， and model， with 10 rats in each group. The general state of all rats was observed. Hematoxylin-eosin （HE） staining was employed to observe the pathological changes of the gastric mucosal tissue. The ultrastructure of cells was observed under transmission electron microscopy. Biochemical methods were adopted to determine the levels of malondialdehyde （MDA） and total superoxide dismutase （T-SOD） in the rat serum. The mRNA and protein levels of Kelch-like ECH-associated protein 1 （Keap1）， Nrf2， NQO1， and HO-1 in the gastric mucosal tissue of rats from each group were measured by Real-time polymerase chain reaction （Real-time PCR） and Western blot， respectively. Reactive oxygen species （ROS） staining was employed to detect the fluorescence intensity of ROS in the gastric mucosal tissue of rats in each group. ResultsCompared with the normal group， the modeled rats were sluggish， had yellowing and sparse hair， short and dark tail， loose stool， and reduced food intake， and showed the gastric mucosa characterized by atrophy， thinning， congestive edema， declined gland density and disarrangement， and apparent inflammatory cell infiltration， irregular nucleus morphology， nuclear condensation， mitochondria with decreased number， declined density， and abnormal morphology， vacuolization， declined serum T-SOD level （P<0.05）， elevated serum MDA level （P<0.05）， downregulated mRNA and protein levels of Nrf2， NQO1， and HO-1 （P<0.05）， upregulated mRNA and protein levels of Keap1 （P<0.05）， and enhanced mean fluorescence intensity of ROS （P<0.05）. Compared with the model group， each treatment group exhibited improved general state， alleviated histopathological changes in gastric mucosa， improved ultrastructural characteristics of cells， elevated T-SOD level and declined MDA level in the serum （P<0.05）， upregulated mRNA and protein levels of Nrf2， NQO1， and HO-1， downregulated mRNA and protein levels of Keap1 （P<0.05）， and attenuated mean fluorescence intensity of ROS （P<0.05）. ConclusionXianglian Huazhuo prescription can improve the general status， ameliorate pathological changes of gastric mucosa， and repair the cell ultrastructure to varying degrees， demonstrating a definite therapeutic effect on CAG. It may restrain OS and safeguard the natural physiological function of gastric mucosa in CAG rats by regulating the Nrf2/NQO1/HO-1 signaling pathway.

Objective To observe the effects of electroacupuncture(EA)on TGF-β1/Smads signaling pathway and epithelial-mesenchymal transition(EMT)related proteins in rats with neurogenic bladder(NB)after spinal cord injury;To explore the possible mechanism of EA in improving NB fibrosis.Methods Totally 36 female SD rats were randomly selected,with 10 rats in the sham-operation group and the remaining 26 rats undergoing complete transection of the spinal cord beneath the T8 vertebrae to establish a NB rat model.The modeling rats were randomly divided into model group and EA group,with 10 rats in each group.EA group was applied to"Ciliao","Zhongji"and"Sanyinjiao",20 min per time,once a day for 7 days.The general condition of the rats in each group was observed,the ultrasound index of the bladder was detected by ultrasound technique,the bladder function of the rats was detected by urodynamics,the body mass of the rats and the wet weight of the bladder were recorded,and the bladder index was calculated.HE staining was used to observe bladder tissue morphology,the degree of bladder tissue fibrosis and bladder wall thickness were detected by Masson staining.The positive expressions of E-cadherin,N-cadherin and Vimentin in bladder tissue were detected by immunohistochemistry.The protein expressions of TGF-β1,p-Smad2/3,E-cadherin,N-cadherin and Vimentin were detected by Western blot.Results Compared with the sham-operation group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the model group significantly increased(P<0.001),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly increased(P<0.001),the bladder index increased significantly(P<0.001),the bladder epithelial cells were thickened and arranged irregularly,the bladder collagen volume fraction and bladder wall thickness significantly increased(P<0.001),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue increased(P<0.01),and the expression of E-cadherin decreased(P<0.001).Compared with the model group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the EA group decreased significantly(P<0.05),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly decreased(P<0.05),the bladder index significantly decreased(P<0.05),the thickness of the bladder epithelial cell layer became thinner and arranged more neatly,and the bladder collagen volume fraction and bladder wall thickness of the bladder were significantly reduced(P<0.05),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue significantly decreased(P<0.05),and the expression of E-cadherin significantly increased(P<0.05).Conclusion EA may reduce the EMT of bladder epithelial cells and decrease the degree of bladder tissue fibrosis by inhibiting the activation of the TGF-β1/Smads signaling pathway,thereby improving bladder function in NB rats after spinal cord injury.

Objective:To evaluate the accuracy and feasibility of an automated scanning and uptake system to assist pathologists with hu-man epidermal growth factor receptor 2(HER2)FISH interpretation.Methods:HER2 gene amplification is detected using FISH,and"result interpretation by independent pathologists"is regarded as the"gold standard."The consistency of"human-machine dialogue results"(use of a CytoVision* system combined with manual interpretation)and"CytoVision*-based automated interpretation"with the"gold standard"was assessed.Results:Consistency between"human-machine dialogue results"and the"gold standard"can surpass 91%,with the former method saving up to 50%of the manual operation time.The tendency of each cell nucleus's HER2 copy number to be"underestimated"is the main reason for the low sensitivity observed in cases with low copy number amplification and HER2 heterogeneous expression cases in"human-machine dialogue interpretation."Conclusions:Automatic FISH image analysis and uptake systems simulate the process of manu-ally interpreted cell selection,ensure random cell selection,and improve work efficiency.With its accurate selection of the hybridization re-gion and"human-computer dialogue,"the system is expected to"replace"interpretation by independent pathologists.

Objective To observe the effects of electroacupuncture(EA)on TGF-β1/Smads signaling pathway and epithelial-mesenchymal transition(EMT)related proteins in rats with neurogenic bladder(NB)after spinal cord injury;To explore the possible mechanism of EA in improving NB fibrosis.Methods Totally 36 female SD rats were randomly selected,with 10 rats in the sham-operation group and the remaining 26 rats undergoing complete transection of the spinal cord beneath the T8 vertebrae to establish a NB rat model.The modeling rats were randomly divided into model group and EA group,with 10 rats in each group.EA group was applied to"Ciliao","Zhongji"and"Sanyinjiao",20 min per time,once a day for 7 days.The general condition of the rats in each group was observed,the ultrasound index of the bladder was detected by ultrasound technique,the bladder function of the rats was detected by urodynamics,the body mass of the rats and the wet weight of the bladder were recorded,and the bladder index was calculated.HE staining was used to observe bladder tissue morphology,the degree of bladder tissue fibrosis and bladder wall thickness were detected by Masson staining.The positive expressions of E-cadherin,N-cadherin and Vimentin in bladder tissue were detected by immunohistochemistry.The protein expressions of TGF-β1,p-Smad2/3,E-cadherin,N-cadherin and Vimentin were detected by Western blot.Results Compared with the sham-operation group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the model group significantly increased(P<0.001),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly increased(P<0.001),the bladder index increased significantly(P<0.001),the bladder epithelial cells were thickened and arranged irregularly,the bladder collagen volume fraction and bladder wall thickness significantly increased(P<0.001),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue increased(P<0.01),and the expression of E-cadherin decreased(P<0.001).Compared with the model group,the upper and lower diameter,anteroposterior diameter,transverse diameter,bladder volume of the EA group decreased significantly(P<0.05),the maximum bladder pressure,leak point pressure difference,perfusion time and maximum bladder capacity significantly decreased(P<0.05),the bladder index significantly decreased(P<0.05),the thickness of the bladder epithelial cell layer became thinner and arranged more neatly,and the bladder collagen volume fraction and bladder wall thickness of the bladder were significantly reduced(P<0.05),the expressions of TGF-β1,p-Smad2/3,N-cadherin and Vimentin in bladder tissue significantly decreased(P<0.05),and the expression of E-cadherin significantly increased(P<0.05).Conclusion EA may reduce the EMT of bladder epithelial cells and decrease the degree of bladder tissue fibrosis by inhibiting the activation of the TGF-β1/Smads signaling pathway,thereby improving bladder function in NB rats after spinal cord injury.

Objective:To evaluate the accuracy and feasibility of an automated scanning and uptake system to assist pathologists with hu-man epidermal growth factor receptor 2(HER2)FISH interpretation.Methods:HER2 gene amplification is detected using FISH,and"result interpretation by independent pathologists"is regarded as the"gold standard."The consistency of"human-machine dialogue results"(use of a CytoVision* system combined with manual interpretation)and"CytoVision*-based automated interpretation"with the"gold standard"was assessed.Results:Consistency between"human-machine dialogue results"and the"gold standard"can surpass 91%,with the former method saving up to 50%of the manual operation time.The tendency of each cell nucleus's HER2 copy number to be"underestimated"is the main reason for the low sensitivity observed in cases with low copy number amplification and HER2 heterogeneous expression cases in"human-machine dialogue interpretation."Conclusions:Automatic FISH image analysis and uptake systems simulate the process of manu-ally interpreted cell selection,ensure random cell selection,and improve work efficiency.With its accurate selection of the hybridization re-gion and"human-computer dialogue,"the system is expected to"replace"interpretation by independent pathologists.

Objective:To evaluate the role of tumor necrosis factor-α-induced protein 8-like molecule-2 (TIPE2) in the endogenous protective mechanism against acute lung injury in septic mice and the relationship with ferroptosis.Methods:Twenty SPF healthy wild-type male C57BL/6N mice and 20 TIPE2 gene knockout C57BL/6N mice, aged 6-8 weeks, weighing 20-25 g, were assigned to wild-type sham operation group (WT-Sham group), wild-type sepsis group (WT-SEP group), TIPE2 knockout sham operation group (KO-Sham group), and TIPE2 knockout sepsis group (KO-SEP group) using a random number table method, with 10 mice in each group. Acute lung injury was induced by cecal ligation and perforation in anesthetized mice. The animals were sacrificed after anesthesia at 24 h after operation and lung tissues were obtained for examination of the morphological results of lung tissues (with a light microscope) and for determination of wet to dry lung weight (W/D) ratio, contents of ferrous ions (Fe 2+ ), glutathione (GSH) and malondialdehyde (MDA) and expression of solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4) and Acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot). Results:Compared with WT-Sham group, the lung injury score, W/D ratio and contents of Fe 2+ and MDA were significantly increased, the content of GSH was decreased, the expression of GPX4 and SLC7A11 was down-regulated, and the expression of ACSL4 was up-regulated in WT-SEP group ( P<0.05). Compared with WT-SEP group, the lung injury score, W/D ratio and contents of Fe 2+ and MDA were significantly increased, the content of GSH was decreased, the expression of GPX4 and SLC7A11 was down-regulated, and the expression of ACSL4 was up-regulated in KO-SEP group ( P<0.05). Conclusions:TIPE2 is involved in the endogenous protective mechanism against acute lung injury, which may be related to the inhibition of ferroptosis in lung tissues of septic mice.

Objective To investigate the temporal and spatial relationship between autophagosome formation during spermatid deformation and lipid droplet distribution in seminiferous tubules,thereby providing insights into the en-ergy sources underlying spermiogenesis.Methods Healthy adult wild-type male mice were used in this study.The expression profile of the autophagy marker microtubule-associated protein light chain 3(LC3)in the testis was ex-amined using immunohistochemical staining and Western blot.The ultrastructural features of autophagosomes were observed via transmission electron microscopy.Lipid droplets in the seminiferous tubules were visualized by Oil Red O staining,and the relationship between lipid metabolism and energy dynamics was assessed by measuring ATP content and ATPase activity.Results Autophagosomes were detected in deforming spermatids and Sertoli cells.LC3 was predominantly expressed in the cytoplasm of elongating spermatids,with increasing levels found from stages Ⅱ to Ⅳ.Concurrently,lipid droplets within these cells also increased,peaking in spermatids and residual bodies during stagesⅦ-Ⅷ,followed by a decline.In contrast,lipid droplets in Sertoli cells remained markedly in-creased during stages Ⅸ-Ⅰ and low during stages Ⅱ-Ⅷ.The change of ATP levels in the seminiferous tubules showed a similar pattern as Sertoli cell lipid content,whereas ATPase activity showed an inverse trend.All these changes displayed a clear stage-dependent manner.Conclusions During spermatid elongation in mice LC3 expres-sion,autophagosome formation,lipid droplet accumulation in both spermatozoa and Sertoli cells,as well as ATP and ATPase levels in seminiferous tubules exhibit a tightly coordinated spatiotemporal relationship.These findings suggest that autophagy and lipid metabolism synergistically contribute to the energy supply required for the extensive cellular remodeling that occurs during spermiogenesis.