OBJECTIVE To investigate the molecular mechanism of Tiaozhong huashi decoction in improving gastroesophageal reflux disease （GERD） by regulating endoplasmic reticulum autophagy in rat esophageal interstitial cells of Cajal （ICC） through p62 signaling pathway.METHODS After establishing a rat GERD model via the weak acid intragastric administration and irregular diet， the rats were randomly divided into model group， western medicine group [1.56 mg/（kg·d） of mosapride， 3.125 mg/（kg·d） of lansoprazole] and low-， medium- and high-dose Tiaozhong huashi decoction [4.713， 9.425， 18.85 g/（kg·d）] groups. A blank group was also set up， with 8 rats in each group. The rats in each group were intragastrically administered with corresponding liquid or normal saline twice a day for 14 days. During the intervention period， the general conditions of rats’ hair， spirit， diet and defecation were observed. After the last gavage， the pathological changes of esophageal tissue and the ultrastructure of ICC were observed. The relative expression and positive expression levels of p62， autophagy-related protein 8 （ATG8）， 200 kDa FAK family kinase-interacting protein （FIP200） and Unc-51 like autophagy activating kinase 1 （ULK1） in esophageal tissue were detected.RESULTS Compared with the blank group， rats in the model group exhibited listlessness； thickening of the esophageal basal layer， elongation of lamina propria papillae with inflammatory cell infiltration； markedly dilated endoplasmic reticulum appearing as round vacuolar cisternae with ribosome detachment； and a small number of accumulated autophagosomes. The relative expression and positive expression levels of p62， ATG8， FIP200， and ULK1 were significantly decreased （ P ＜0.05）. Compared with the model group， the above pathological injuries in the medium- and high-dose Tiaozhong huashi decoction groups had all improved to varying degrees； the damaged endoplasmic reticulum structure was repaired， and no autophagosome accumulation was observed. In particular， the relative expression levels and positive expression levels of p62， ATG8， FIP200， and ULK1 in esophageal tissue of rats in the medium-dose Tiaozhong huashi decoction group were significantly increased （ P ＜0.05）.CONCLUSIONS Tiaozhong huashi decoction ameliorated esophageal mucosal injury in GERD rats， and its mechanism was possibly associated with activation of the p62 signaling pathway and restoration of the endoplasmic reticulum structure， morphology， and autophagic capacity of ICC.

Liver cirrhosis as the terminal stage of chronic liver disease has seen many new insights and advances in its treatment strategies and perspectives in recent years. However, there are still many controversies about cirrhotic portal hypertension management, prevention, therapy, and complications. This article summarizes the main key controversial points in the current treatment of liver cirrhosis from an evidence-based medicine perspective, including the use of non-selective β-blockers during decompensated stages, exploration of precise strategies for albumin, re-evaluation of the risks of statins, weighing the pros and cons of proton pump inhibitors, new understandings of anticoagulation therapy, breakthroughs in targeting gut microbiota, and nutritional support management. In addition, it combines the latest research data and guideline recommendations to explore future development directions so as to provide clinical practice reference.

Objective To evaluate the therapeutic efficacy of stenting combined with medication versus medication alone in treating severe extracranial carotid artery stenosis.Methods This prospective,multicenter cohort study enrolled 337 patients with carotid artery stenosis ranging from 70%to 99%.Based on patient preference,they were divided into two groups:a stent combined with medication group(antiplatelet drugs+statins,n=169)and a medication-only group(n=168).The study assessed perioperative safety and the incidence of recurrent cerebrovascular events related to the target vessel within one year.Results The success rate of the stent combined with medication group was 100%.There was one case of hyperperfusion-related intracranial hemorrhage during the perioperative period(1/169)The patient recovered well after medical treatment.There was no statistically significant difference in perioperative complications between the two groups(P=0.318).During the 1-year follow-up,the stent combined with medication group(3/169)had a significantly lower incidence of recurrent ischemic stroke compared to the medication-only group(15/168),with a statistically significant difference(P=0.003).Univariate analysis showed that gender(χ2=4.372,P=0.037),stenosis rate≥85%(χ2=10.738,P=0.001),and non-stent treatment(χ2=7.171,P=0.007)were risk factors for recurrent stroke.However,the number of recurrent ischemic strokes in females was relatively small and after adjusting the chi-square test,the difference was not statistically significant(P=0.075).Logistic regression analysis revealed that stenosis rate≥85%(OR=115.925,95%CI:2.077-122.102,P=0.008)and non-stent treatment(OR=0.158,95%CI:0.044-0.564,P=0.004)were independent risk factors for recurrent cerebrovascular events within one year.Conclusions In patients with≥85%extracranial carotid artery stenosis,stenting combined with medication reduces recurrent ischemic cerebrovascular events within 1 year.

BACKGROUND:The micro/nanostructured gradient biomimetic surface of implant materials can simulate the structure of the extracellular environment in human bone tissue,thereby achieving perfect bone integration function.However,further research is needed on the mechanisms by which the surface microstructure of bone implant materials regulates cell function and promotes osteogenesis. OBJECTIVE:To analyze the effect of titanium sheet microstructure surface on osteogenic differentiation of MC3T3-E1 osteoblasts. METHODS:(1)At a constant voltage of 5 V or 20 V,nanotube arrays of different diameters were prepared on the surface of titanium sheets by acid etching and anodic oxidation techniques,and were recorded as group R5 and group R20,respectively.The surface morphology,roughness,and hydrophilicity of pure titanium sheet(without acid etching or anodizing treatment)were measured in group R5 and group R20.(2)MC3T3-E1 osteoblasts of logarithmic growth stage were inoculated on the surface of pure titanium sheets,R5 group and R20 group respectively.After 24 hours of osteogenic induction culture,the expression of mechanical sensitive channel protein 1 was analyzed by RT-PCR and immunofluorescence staining.Osteoblast inducible base with or without the mechanosensitive channel protein 1 activator Yada1 was added,and alkaline phosphatase staining was performed after 7 days of culture.Alizarin red staining was performed after 14 days of culture. RESULTS AND CONCLUSION:(1)The surface of pure titanium sheets was smooth under scanning electron microscope.Relatively uniform and orderly nanotube arrays with average diameters of about 30 nm and 100 nm were observed on the surface of titanium sheets of groups R5 and R20,respectively.The results of scanning electron microscope were further verified by atomic force microscopy.The surface roughness of titanium sheet of group R5 was higher than that of pure titanium(P<0.05),and the water contact angle was lower than that of pure titanium(P<0.05).The surface roughness of titanium sheet in group R20 was higher than that in group R5(P<0.05),and the water contact angle was lower than that in group R5(P<0.05).(2)RT-PCR and immunofluorescence staining showed that the expression of mechanosensitive channel protein 1 in group R5 was higher than that in pure titanium group(P<0.05),and the expression of mechanosensitive channel protein 1 in group R20 was higher than that in group R5(P<0.05).Under the osteogenic induction,compared with the condition without Yada1,there were no significant changes in the activity of alkaline phosphatase and the deposition of calcified nodules in pure titanium group after Yada1 addition,while the activity of alkaline phosphatase and the deposition of calcified nodules in groups R5 and R20 after Yada1 addition were significantly increased(P<0.05).With or without Yada1,the alkaline phosphatase activity and calcified nodule deposition in group R5 were higher than those in pure titanium group(P<0.05),and the alkaline phosphatase activity and calcified nodule deposition in group R20 were higher than those in group R5(P<0.05).(3)The results show that the surface microstructure of titanium sheet can promote the osteogenic differentiation of osteoblast MC3T3-E1 by activating mechanosensitive channel protein 1.

Liver cirrhosis as the terminal stage of chronic liver disease has seen many new insights and advances in its treatment strategies and perspectives in recent years. However, there are still many controversies about cirrhotic portal hypertension management, prevention, therapy, and complications. This article summarizes the main key controversial points in the current treatment of liver cirrhosis from an evidence-based medicine perspective, including the use of non-selective β-blockers during decompensated stages, exploration of precise strategies for albumin, re-evaluation of the risks of statins, weighing the pros and cons of proton pump inhibitors, new understandings of anticoagulation therapy, breakthroughs in targeting gut microbiota, and nutritional support management. In addition, it combines the latest research data and guideline recommendations to explore future development directions so as to provide clinical practice reference.

Objective To evaluate the therapeutic efficacy of stenting combined with medication versus medication alone in treating severe extracranial carotid artery stenosis.Methods This prospective,multicenter cohort study enrolled 337 patients with carotid artery stenosis ranging from 70%to 99%.Based on patient preference,they were divided into two groups:a stent combined with medication group(antiplatelet drugs+statins,n=169)and a medication-only group(n=168).The study assessed perioperative safety and the incidence of recurrent cerebrovascular events related to the target vessel within one year.Results The success rate of the stent combined with medication group was 100%.There was one case of hyperperfusion-related intracranial hemorrhage during the perioperative period(1/169)The patient recovered well after medical treatment.There was no statistically significant difference in perioperative complications between the two groups(P=0.318).During the 1-year follow-up,the stent combined with medication group(3/169)had a significantly lower incidence of recurrent ischemic stroke compared to the medication-only group(15/168),with a statistically significant difference(P=0.003).Univariate analysis showed that gender(χ2=4.372,P=0.037),stenosis rate≥85%(χ2=10.738,P=0.001),and non-stent treatment(χ2=7.171,P=0.007)were risk factors for recurrent stroke.However,the number of recurrent ischemic strokes in females was relatively small and after adjusting the chi-square test,the difference was not statistically significant(P=0.075).Logistic regression analysis revealed that stenosis rate≥85%(OR=115.925,95%CI:2.077-122.102,P=0.008)and non-stent treatment(OR=0.158,95%CI:0.044-0.564,P=0.004)were independent risk factors for recurrent cerebrovascular events within one year.Conclusions In patients with≥85%extracranial carotid artery stenosis,stenting combined with medication reduces recurrent ischemic cerebrovascular events within 1 year.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective:To construct and verify a prognosis model of nomogram for the patients with locally advanced oral squamous cell carcinoma(OSCC).Methods:7 118 clinical cases with locally advanced OSCC from 2010 to 2015 in SEER database were in-cluded and randomly divided into training set(TS)and validation set(VS)by a 7:3 ratio.In TS,univariate and multivariate Cox proportional risk models were used to screen and determine the prognostic factors of locally advanced OSCC,and a nomogram predic-tion model was constructed to predict the cancer specific survival rate(CSS)of patients in 3 and 5 years.In TS and VS,the consis-tency index(C-index)and ROC curves were used to evaluate the differentiation of the model,the calibration curves were used to eval-uate the prediction accuracy of the model,and the decision curve analysis(DC A)method was used to evaluate the clinical applicabil-ity of the model.Results:4 984 and 2 134 cases were respectively included in TS and VS.No statistical difference of the analysis in-dexes was found between the 2 sets(P＞0.05).10 risk factors affecting the prognosis of the patients with locally advanced OSCC were identified in TS,including age,sex,race,marital status,location of onset,degree of histological differentiation,T stage,N stage,with or without operation and radiotherapy(P＜0.05).In the TS and VS,the C-index of the nomogram was 0.702 and 0.704 respectively,and the AUC of the 3-year and 5-year CSS was between 0.727 and 0.736.The calibra-tion curves showed that the 3-and 5-year CSS predicted by the nomogram model was in good agreement with the actual CSS.The DCA curves showed that the clinical benefit of the nomogram was better than that of the 7th e-dition of the AJCC TNM staging system.Conclusion:The nomogram constructed in this study can accurately predict the survival rate of OSCC patients with locally advanced tumor.