[Objective]To investigate the effect of nicotinamide adenine dinucleotide on vascular endothelial injury in hypertension and its molecular mechanism.[Methods]C57BL/6J mice were randomly divided into saline group(Saline)and hypertension group(Ang Ⅱ,which were infused with Ang Ⅱ via subcutaneously implanted osmotic pumps),and supplemented daily with nicotinamide mononucleotide(300 mg/kg),a precursor of NAD+.Blood pressure,endothelial relaxation function and pulse wave velocity were measured after 4 weeks.Wound healing assay and adhesion assay were used to evaluate the function of endothelial cells in vitro.mtROS levels were detected by immunofluorescence staining.RT-PCR was used to detect the mRNA expression of mtDNA,SIRT3 and isocitrate dehydrogenase 2(IDH2).8-hydroxy-2'-deoxyguanosine levels were detected by enzyme-linked immunosorbent assay.The protein expression levels of p-eNOS,eNOS,SIRT3 and IDH2 were detected by Western blot.[Results]NMN supplementation reduced blood pressure(P<0.001)and improved endothelial function and arterial stiffness(P<0.001)in hypertensive mice.In vitro,NMN improved endothelial function in Ang Ⅱ-stimulated endothelial cells(P<0.05)and attenuated mitochondrial oxidative stress levels(P<0.001).Mechanistically,NMN elevated SIRT3 activity(P<0.001),which subsequently enhanced IDH activity(P<0.001)and reduced oxidative stress levels in endothelial cells.Conversely,knockdown of IDH2 would reverse the effect of SIRT3 in improving endothelial function(P<0.001).[Conclusion]NAD+lowers blood pressure and enhances vascular function in hypertension by reducing the level of oxidative stress in endothelial cells through activation of the SIRT3/IDH2 signal pathway.

Objective To explore the correlation between the expression of serum microRNA(miR)-1298-5p,miR-625-5p,and miR-155 with the degree of Helicobacter pylori(Hp)infection in elderly gastric cancer patients.Methods From January 2021 to November 2023,120 elderly patients with gastric cancer admitted to the hospital from January 2021 to November 2023 were selected as the gastric cancer group,and 130 non-gas-tric cancer patients who underwent gastroscopy were selected as the control group.The expression levels of miR-1298-5p,miR-625-5p and miR-155 in serum were detected by fluorescence quantitative PCR(qPCR).Car-bon 13 urea breath test was used to detect the positive rate of Hp infection in two groups,and the degree of Hp infection in elderly patients with gastric cancer were evaluated.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of serum miR-1298-5p,miR-625-5p,and miR-155 expression levels for Hp infection in elderly gastric cancer patients.Pearson method was applied to analyze the correlation between serum miR-1298-5p,miR-625-5p,miR-155 expression and positive rate of Hp infection in elderly gas-tric cancer patients.Results Compared with the control group,the expression levels of miR-1298-5p and miR-625-5p in serum of gastric cancer group decreased(P＜0.05),while the positive rate of Hp infection and the expression level of serum miR-155 increased(P＜0.05).The expression levels of serum miR-1298-5p and miR-625-5p in elderly gastric cancer patients with Hp grade Ⅰ,Ⅱ,and Ⅲ infection were lower than those without Hp infection,while the expression level of miR-155 was higher(P＜0.05).Patients with poor differ-entiation,lymph node metastasis,and TNM stage Ⅲ-Ⅳ had lower expressions of serum miR-1298-5p and miR-625-5p(P＜0.05),and higher expression of miR-155(P＜0.05)than those with moderate-high differen-tiation,no lymph node metastasis,and TNM stage Ⅰ-Ⅱ.The expression levels of serum miR-1298-5p and miR-625-5p were negatively correlated with the positive rate of Hp infection in elderly patients with gastric cancer(r=-0.443,-0.386,both P＜0.001),and the expression levels of serum miR-155 were positively correlated with the positive rate of Hp infection(r=0.525,P＜0.001).The area under the curve(AUC)of serum miR-1298-5p,miR-625-5p and miR-155 combined diagnosis of Hp infection in elderly gastric cancer pa-tients was higher than that of single diagnosis(P＜0.05).Conclusion The expression levels of miR-1298-5p and miR-625-5p in serum of elderly gastric cancer patients with Hp infection decrease,while the expression level of miR-155 increases.These three factors are related to the degree of Hp infection and have good diag-nostic value for the occurrence of Hp infection.

OBJECTIVES: To investigate the molecular mechanism by which salvianolic acid B (Sal-B) modulates mitochondrial functional homeostasis and alleviates myocardial ischemia-reperfusion (I/R) injury in mice. METHODS: Mouse cardiomyocyte HL-1 cells were pretreated with 5 μmol/L Sal-B with or without sh-Sirt1 transfection before exposure to hypoxia-reoxygenation (HR), and the changes in ATP production, mitochondrial superoxide activity, substrate oxidation level were evaluated. In the animal experiment, 36 C57BL/6J mice were randomized into 3 groups (n=12) for sham operation or ligation of the left anterior coronary artery to induce myocardial I/R injury with or without intravenous injection of Sal-B+I/R (50 mg/kg). In the rescue experiment, 60 adult C57BL/6J mice were randomized into 5 groups (n=12): sham-operated group, myocardial I/R group, Sal-B+I/R group, I/R+Sal-B+Sirt1fl/fl group, and I/R+Sal-B+cKO-Sirt1 group. Myocardial injury was evaluated with HE staining, and cardiac function was assessed by measurement of the ejection fraction and fractional shortening using echocardiography. RESULTS: In HL-1 cells with HR injury, Sal-B pretreatment significantly increased cellular ATP production, reduced mitochondrial superoxide anion levels, and enhanced oxygen consumption level. In the mouse models of myocardial I/R injury, Sal-B pretreatment markedly ameliorated I/R-induced structural disarray of the cardiac myocytes and improved cardiac ejection. Cycloheximide chase with Western blotting and ubiquitination assays after Sirt1-IP showed that Sal-B significantly inhibited Sirt1 degradation in HL-1 cells. Sirt1 knock-down reversed Sal-B-induced increases in ATP production, reduction in superoxide, and elevation of OCR in HL-1 cells. Cardiomyocyte-specific Sirt1 knockout obviously reversed Sal-B-mediated improvement in cardiac ejection function and myocardial structure damage in mice with myocardial I/R injury. CONCLUSIONS Sal-B promotes mitochondrial functional homeostasis in cardiomyocytes with HR injury and improves cardiac function in mice after myocardial I/R by inhibiting Sirt1 protein degradation.
Animals ; Sirtuin 1/metabolism* ; Myocardial Reperfusion Injury/physiopathology* ; Mice, Inbred C57BL ; Mice ; Myocytes, Cardiac/drug effects* ; Benzofurans/pharmacology* ; Homeostasis/drug effects* ; Male ; Mitochondria/drug effects* ; Depsides

ObjectiveTo investigate the accuracy of multiple discriminant analysis （MDA） versus fibrosis-4 （FIB-4） in assessing liver fibrosis degree in patients with HBV infection， as well as the possibility of MDA as an indicator for disease progression. MethodsA total of 263 patients with HBV infection who underwent liver biopsy in The First Affiliated Hospital of Guangxi Medical University from April 2010 to April 2024 were included， and their clinical data were collected. According to the results of pathological examination， they were divided into non-significant fibrosis group （F<2） with 126 patients and significant fibrosis group （F≥2） with 137 patients. The correlation of MDA and FIB-4 with liver fibrosis degree was analyzed， and MDA and FIB-4 were compared in terms of their accuracy in assessing significant liver fibrosis. A total of 62 patients completed follow-up， and according to the presence or absence of progression to liver cirrhosis at the last follow-up visit， they were divided into progressive group with 21 patients and non-progressive group with 41 patients； the efficacy of MDA and FIB-4 in diagnosing disease progression was analyzed and compared. The independent-samples t test was used for comparison of normally distributed continuous data between groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups； the Kruskal-Wallis H test was used for comparison between multiple groups， and the Bonferroni method was used for further comparison between two groups. The chi-square test was used for comparison of categorical data. The Spearman’s correlation coefficient was used for correlation analysis. The Wilcoxon signed rank sum test was used for the analysis of baseline data and data at the end of follow-up， and the binary Logistic regression analysis was used to investigate the influencing factors for progression to liver cirrhosis. The receiver operating characteristic （ROC） curve was used to investigate the diagnostic efficacy of indicators， the Z-test was used for comparison of the area under the ROC curve （AUC）， and the paired chi-square test was used for comparison of the sensitivity， specificity， and accuracy of the two indicators. ResultsThe correlation coefficient between FIB-4 and liver fibrosis degree was 0.378， while the correlation coefficient between MDA and liver fibrosis degree was -0.325 （both P<0.001）. FIB-4 had an AUC of 0.688， a sensitivity of 64.96%， a specificity of 68.87%， a positive predictive value of 67.42%， a negative predictive value of 63.36%， an accuracy of 65.40%， and a cut-off value of 1.01， while MDA had an AUC of 0.653， a sensitivity of 52.55%， a specificity of 78.57%， a positive predictive value of 72.73%， a negative predictive value of 60.37%， an accuracy of 65.02%， and a cut-off value of 0.29， suggesting that compared with FIB-4， MDA had a lower sensitivity （P=0.004） and a higher specificity （P=0.001）. The progressive group had a significantly higher age than the non-progressive group at baseline （t=2.611， P=0.011）. For the progressive group， there was an increase in FIB-4 and a reduction in MDA from baseline to the end of follow-up （both P<0.001）， while the non-progressive group showed no significant changes （both P>0.05）. The multivariate Logistic regression analysis showed that aspartate aminotransferase （odds ratio ［OR］=0.940， 95% confidence interval ［CI］： 0.885‍ ‍—‍ ‍0.998， P<0.05） and MDA （OR=0.445， 95%CI： 0.279‍ ‍—‍ ‍0.710， P<0.001） were independent influencing factors for disease progression. MDA had an AUC of 0.893 and an optimal cut-off value of -0.01 in diagnosing the disease progression of liver cirrhosis. ConclusionMDA has a comparable accuracy to FIB-4 in the diagnosis of significant liver fibrosis， and MDA<-0.01 has a high accuracy in diagnosing the progression of liver fibrosis to liver cirrhosis， which can help to reduce the need for liver biopsy in clinical practice.

Objective:To investigate the relationship between the expression level of the stem cell gene LGR5 and the clinical characteristics and prognosis of colon cancer. Methods:This study included 585 patients with colon cancer, whose clinical data and transcriptome sequencing data were derived from the GSE39582 dataset in the Gene Expression Omnibus (GEO). Among the 585 patients, 322 were male and 263 were female, with ages ranging from 24 to 96 years with a mean age of (67±13) years. According to the TNM staging system, 38 patients were classified as stage Ⅰ, 271 as stage Ⅱ, 210 as stage Ⅲ, and 60 as stage Ⅳ, with staging information missing for 6 patients. The expression level of LGR5 in tumor tissues was detected using gene sequencing technology, and patients were divided into high-expression group (expression level≥7.306, n=292) and low-expression group (expression level<7.306, n=293) based on the median expression level of LGR5(7.306). Clinical characteristics and molecular features between the two groups were compared using t-tests or χ2 tests. Kaplan-Meier survival curves and the log-rank test were used to analyze survival data, including comparisons of overall survival (OS) and disease-free survival (DFS). Results:The expression level of LGR5 was not significantly correlated with the TNM staging of colon cancer but was closely related to the molecular biological features of the tumor. In the high-expression group of LGR5, tumors were more frequently located in the distal colon, with higher proportions of pMMR, CIMP -, CIN +, and BRAF wild type, higher tumor cell purity, and less immune cell infiltration. Survival analysis showed that the overall survival (OS) of the high-expression group was significantly better than that of the low-expression group ( P=0.008 4), with median survival times of 170 months and 96 months, respectively; however, there was no significant difference in disease-free survival (DFS) between the two groups ( P=0.36). Variables that showed significant differences between the two groups were identified and subjected to subgroup analysis. Subgroup analysis indicated that in colon cancer through the traditional adenoma-carcinoma pathway, high expression of LGR5 was associated with better overall survival, while in colon cancer through other molecular biological pathways, the expression level of LGR5 was not significantly related to prognosis. Conclusions:The expression level of LGR5 is significantly correlated with the molecular biological features of colon cancer and serves as a favorable prognostic indicator in colon cancer through the traditional adenoma-carcinoma pathway. LGR5 may be involved in the occurrence, development, and prognosis of colon cancer by affecting the tumor microenvironment and tumor cell characteristics.

Objective:To identify risk factors influencing clinical efficacy of endoscopic resection of small rectal neuroendocrine tumor (NETs).Methods:A retrospective analysis was conducted on patients with rectal NETs ≤10 mm who underwent endoscopic resection from 2013 to 2022. Patients were divided into the endoscopic submucosal dissection (ESD) group and the endoscopic mucosal resection with ligation (EMRL) group according to the treatment methods. After comparing the baseline data, propensity score matching was performed to compare the rates of R1 resection and adequacy of vertical margin distance.Results:A total of 186 patients were included in this study, with 139 receiving ESD and 47 receiving EMRL. The R1 resection rates were 12.2% (17/139) and 2.1% (1/47) in the ESD and EMRL group, respectively ( χ2=3.027, P=0.082). A significant difference in vertical margin adequacy was observed between the two groups [69.1% (96/139) VS 85.1% (40/47), χ2=4.598, P=0.032]. After propensity score matching, 46 pairs of cases were included, and there were no significant differences in the R1 resection rate [6.5% (3/46) VS 2.2% (1/46), χ2=0.261, P=0.609] and vertical margin adequacy [78.3% (36/46) VS 84.8% (39/46), χ2=0.649, P=0.420] between the two groups. Univariate and multivariate Logistic regression analyses revealed that operator experience and preoperative biopsy were independent risk factors for inadequate margin. Conclusion:Treatment method may not be the key factor affecting the distance of the vertical margin after endoscopic resection, but preoperative biopsy and operator experience have a significant impact on margins. Biopsy before endoscopic resection should be avoided, and less experienced doctors are recommended to use EMRL method for small NETs due to its ease of execution.

Objective:To identify risk factors influencing clinical efficacy of endoscopic resection of small rectal neuroendocrine tumor (NETs).Methods:A retrospective analysis was conducted on patients with rectal NETs ≤10 mm who underwent endoscopic resection from 2013 to 2022. Patients were divided into the endoscopic submucosal dissection (ESD) group and the endoscopic mucosal resection with ligation (EMRL) group according to the treatment methods. After comparing the baseline data, propensity score matching was performed to compare the rates of R1 resection and adequacy of vertical margin distance.Results:A total of 186 patients were included in this study, with 139 receiving ESD and 47 receiving EMRL. The R1 resection rates were 12.2% (17/139) and 2.1% (1/47) in the ESD and EMRL group, respectively ( χ2=3.027, P=0.082). A significant difference in vertical margin adequacy was observed between the two groups [69.1% (96/139) VS 85.1% (40/47), χ2=4.598, P=0.032]. After propensity score matching, 46 pairs of cases were included, and there were no significant differences in the R1 resection rate [6.5% (3/46) VS 2.2% (1/46), χ2=0.261, P=0.609] and vertical margin adequacy [78.3% (36/46) VS 84.8% (39/46), χ2=0.649, P=0.420] between the two groups. Univariate and multivariate Logistic regression analyses revealed that operator experience and preoperative biopsy were independent risk factors for inadequate margin. Conclusion:Treatment method may not be the key factor affecting the distance of the vertical margin after endoscopic resection, but preoperative biopsy and operator experience have a significant impact on margins. Biopsy before endoscopic resection should be avoided, and less experienced doctors are recommended to use EMRL method for small NETs due to its ease of execution.

Objective:To investigate the clinical characteristics and prognosis of CpG island methylator phenotype (CIMP+ ) colon cancer, and the significance of CIMP status in the diagnosis and prognosis prediction in defective mismatch repair (dMMR) colon cancer.Methods:The keywords "colorectal cancer" "patient" and "CpG Island Methylator Phenotype" were used to search the Gene Expression Omnibus (GEO) database, and the GSE39582 was obtained, which included the clinical data of 585 patients with colorectal cancer and the sequencing data of the whole transcriptome of the tumor tissues. After excluding 72 cases with missing CIMP values, 513 cases were included for further analysis, including 278 males and 235 females, with a mean age of (67±13) years. According to the CIMP status, they were divided into CIMP+ group ( n=93) and CIMP-group ( n=420), then compare the differences in clinical characteristics, the Kaplan-Meier survival curves were plotted to compare the overall survival and disease-free survival; 71 dMMR cases were divided into CIMP+ group ( n=43) and CIMP-group ( n=28), and the K-M curves were plotted to analyze the differences in overall survival (OS) and disease free survival (DFS). Comparisons between groups were performed by t-test, χ2 test or Mann-Whitney U nonparametric test, and the difference in survival curves was tested by Long-rank test. Results:Patients in the CIMP+ group were significantly older than those in the CIMP-group [(70.84±12.60) years vs (66.21±13.08) years, t=3.18, P=0.002]. Right colon tumors originating from the CIMP+ molecular pathway were 9.3 times more likely to be CIMP+ than those of the left colon cancers ( OR=9.3, 95% CI: 5.2-17.9). BRAF mutant colon cancer originating from CIMP+ was 215.2 times more common than BRAF wild-type colon cancer originating with CIMP+ ( OR=215.2, 95% CI: 53.2-1906.7); and patients with dMMR colon cancer originated 12.8 times more common than patients with pMMR ( OR=12.8, 95% CI: 7.0-23.9). The difference between the CIMP+ and CIMP-groups was not statistically significant in terms of overall survival and disease-free survival ( P=0.590, 0.220). In the dMMR colon cancer subgroup, CIMP status did not correlate with patients′ overall survival and disease-free survival ( P>0.05). Conclusions:CIMP+ colon cancer patients were mostly of advanced age, with tumors originating from the right colon, mostly combined with BRAF gene mutations, and manifested as mismatch repair-deficient colon cancers. CIMP status had no correlation with TNM stage and survival of colon cancers patients. There was no significant difference in the survival between dMMR colon cancers caused by CIMP+ and those caused by MMR gene mutations.

ObjectiveTo investigate the performance of fibrosis-4 （FIB-4） versus aspartate aminotransferase-to-platelet ratio index （APRI） in predicting advanced liver fibrosis and disease progression in patients with chronic HBV infection. MethodsA total of 497 patients with chronic HBV infection who underwent liver biopsy in The First Affiliated Hospital of Guangxi Medical University from February 2013 to December 2022 were enrolled， among whom 404 were enrolled in a retrospective study and 75 were enrolled in a prospective study. Related indicators were collected， including demographic features （sex and age）， biochemical indices （alanine aminotransferase ［ALT］ and aspartate aminotransferase ［AST］）， and platelet count， and FIB-4 and APRI were calculated. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups； the chi-square test was used for comparison of categorical data between groups. The area under the ROC curve （AUC） was used to assess the ability of APRI and FIB-4 in evaluating liver fibrosis degree and disease progression in patients with chronic HBV infection. ResultsIn the retrospective analysis， compared with the FIB-4<2.67 group， the FIB-4≥2.67 group had a significantly higher proportion of the patients who were diagnosed with liver cirrhosis or hepatocellular carcinoma （66.19% vs 47.54%， χ²=12.75， P<0.001）. The medians of FIB-4 and APRI increased significantly with liver fibrosis degree from F0 to F4 （H=42.5 and 35.9， both P<0.001）. As for the fibrosis stage of F0-F4， the median of FIB-4 was significantly higher than that of APRI in the patients with the same fibrosis stage （H=59.71， P<0.001）. FIB-4 and APRI had a similar AUC for predicting stage F3 fibrosis （0.67 vs 0.65， Z=0.71， P=0.480）， while FIB-4 had a higher AUC for predicting stage F4 fibrosis than APRI （0.72 vs 0.64， Z=10.50， P<0.001）. In the prospective study cohort， FIB-4 and APRI showed an increasing trend over time in predicting disease progression （chronic hepatitis B to liver cirrhosis）， with an AUC of 0.718 （95% confidence interval ［CI］： 0.476‍ ‍—‍ ‍0.760） and 0.555 （95%CI： 0.408‍ ‍—‍ ‍0.703）， respectively， and FIB-4 had a significantly higher accuracy than APRI in predicting disease progression （χ²=12.44， P<0.001）. ConclusionFIB-4 and APRI can be used to evaluate advanced liver fibrosis （F3 and F4） and predict disease progression， and FIB-4 is superior to APRI in certain aspects.

Objective:To explore the application effects of a neurosurgical mixed-reality distance teaching (NMDT) model in standardized residency training in neurosurgery.Methods:We built an NMDT system using mixed-reality technology and remote interaction technology, and designed the implementation procedure according to the teaching objectives. After the teaching activities were completed, a teaching satisfaction questionnaire survey was conducted among 20 neurosurgery resident trainees, in which they provided satisfaction scores for the same teaching content with different teaching models (i.e., the NMDT model and traditional teaching model). SPSS 22.0 software was used to perform the t test for data analysis. Results:There were significant differences between the NMDT model and the traditional teaching model in key indicators including the score for "completion of teaching objectives" (9.20±0.68 vs. 8.25±0.70, P<0.001) and the score for "satisfaction with learning gains" score (8.95±0.67 vs. 8.05±0.92, P=0.001). The NMDT model also outperformed the traditional teaching model in the other individual scores and the total score. Conclusions:The NMDT model can improve teaching quality, increase training efficiency, and enrich teaching content, which is worthy of promotion.