ObjectiveTo establish an ultra-performance liquid fingerprint and multi-components determination method for Naomaili granules. To evaluate the quality of different batches by chemometrics， and the anti-neuroinflammatory effects of water extract and main components of Naomaili granules were tested in vitro. MethodsThe similarity and common peaks of 27 batches of Naomaili granules were evaluated by using Ultra performance liquid chromatography （UPLC） fingerprint detection. Ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） technology was used to determine the content of the index components in Naomaili granules and to evaluate the quality of different batches of Naomaili granules by chemometrics. LPS-induced BV-2 cell inflammation model was used to investigate the anti-neuroinflammatory effects of the water extract and main components of Naomaili granules. ResultsThe similarity of fingerprints of 27 batches of samples was > 0.90. A total of 32 common peaks were calibrated， and 23 of them were identified and assigned. In 27 batches of Naomaili granules， the mass fractions of 14 components that were stachydrine hydrochloride， leonurine hydrochloride， calycosin-7-O-glucoside， calycosin，tanshinoneⅠ， cryptotanshinone， tanshinoneⅡ_A， ginsenoside Rb₁， notoginsenoside R₁， ginsenoside Rg₁， paeoniflorin， albiflorin， lactiflorin， and salvianolic acid B were found to be 2.902-3.498， 0.233-0.343， 0.111-0.301， 0.07-0.152， 0.136-0.228， 0.195-0.390， 0.324-0.482， 1.056-1.435， 0.271-0.397， 1.318-1.649， 3.038-4.059， 2.263-3.455， 0.152-0.232， 2.931-3.991 mg∙g^-1， respectively. Multivariate statistical analysis showed that paeoniflorin， ginsenoside Rg₁， ginsenoside Rb₁ and staphylline hydrochloride were quality difference markers to control the stability of the preparation. The results of bioactive experiment showed that the water extract of Naomaili granules and the eight main components with high content in the prescription had a dose-dependent inhibitory effect on the release of NO in the cell supernatant. Among them， salvianolic acid B and ginsenoside Rb₁ had strong anti-inflammatory activity， with IC₅₀ values of （36.11±0.15） mg∙L^-1 and （27.24±0.54） mg∙L^-1， respectively. ConclusionThe quality evaluation method of Naomaili granules established in this study was accurate and reproducible. Four quality difference markers were screened out， and eight key pharmacodynamic substances of Naomaili granules against neuroinflammation were screened out by in vitro cell experiments.

Objective To study the chemical constituents of Fallopia aubertii L.Henry Holub. Methods The chemical constituents of Fallopia aubertii L.Henry Holub. were separated and purified by online two-dimensional preparative liquid chromatography and identified by physical and chemical constants and spectral analysis. The inhibitory activities on xanthine oxidase were determined by ultraviolet spectrophotometry. Results Ten compounds were isolated from the extract of Fallopia aubertii L.Henry Holub, including isotachioside（1）, 3,4,5-trimethoxyphenyl-（6'-O-galloyl）-O-β-D-Glucopyranoside（2）, 1-hydroxy-,4,5-1-O-[6'-O-（4''-carboxy-1'',3'',5'trihydrotrimethoxyphenylxy）-phenyl]-β-D-glucopyranoside（3）, myricetrin（4）, myricetin（5）, rutin（6）, quercetin-3-O-β-D-galactoside（7）, quercetin-3-O-β-D-glucopyranoside（8）, lyciumideA（9）, and N-trans-Feruloyltyramine（10）. The inhibitory activity test results showed that the IC₅₀ of compound 5 was 15.92 μmol/L, and the IC₅₀ of compound 6 was 87.36 μmol/L. Conclusion Compounds 1,2,3,4 and 8 were isolated from Medicago polymorpha for the first time. Compounds 5 and 6 had xanthine oxidase inhibitory activity.

Radical resection of mid-and low-rectal cancer requires not only oncologic safety but also preservation of organs and postoperative bowel function.While a 1-2 cm distal resection margin has been largely accepted,the optimal length of the proximal margin remains highly controversial.Clinically,the"10-cm rule"derived from colon cancer is often referenced,yet its applicability to rectal cancer lacks consistent supporting evidence.Previous studies have shown that an excessively long proximal margin may increase anastomotic tension and lead to anastomotic leakage,whereas insufficient resection heightens the risk of positive margins and local recurrence.In addition,the extent of lymph node metastasis,vascular perfusion of the proximal bowel,radiation-induced injury after neoadjuvant chemoradiotherapy,and postoperative bowel function-particularly low anterior resection syndrome-are all important factors influencing the selection of the proximal margin.In recent years,the application of indocyanine green fluorescence imaging has provided new evidence for intraoperative assessment of bowel perfusion;for patients receiving neoadjuvant chemoradiotherapy,radiation injury presents a gradient pattern,and resecting approximately≥20 cm proximal to the tumor may reduce the incidence of anastomosis-related complications.Based on current literature,this review provides a systematic overview of the historical evolution,influencing factors,and clinical evidence regarding proximal resection margins in rectal cancer surgery,with the aim of informing individualized margin selection and optimizing surgical strategies.

African swine fever(ASF)is an acute,febrile,and highly fatal disease caused by African swine fever virus(ASFV)in pigs.Given the current lack of commercial vaccines and the continu-ous evolution of ASFV in recent years,the emergence of moderately virulent genotype Ⅱ strains and the introduction of genotype Ⅰ attenuated strains have led to persistent and chronic infections in pigs.Therefore,the detection of specific antibodies against ASFV has become imperative.In this study,we established a competitive chemiluminescence immunoassay(p30-cCLIA)for detecting ASFV p30 antibodies using p30 monoclonal antibodies.By detecting sera with clear negative and positive backgrounds,we determined that the Cut-off value of this method was 50％,with both di-agnostic sensitivity(Dsn)and diagnostic specificity(Dsp)reaching 100％.Under optimal reaction conditions,we screened out an enzyme-labeled stabilizer suitable for p30 monoclonal antibody 16-5E7E8-HRP.Furthermore,the sensitivity of the established p30-cCLIA method was higher than that of the commercial blocking ELISA kit(1∶2 048 vs 1∶512)and exhibited good repeatability.Detection of sera positive for other porcine virus infections showed no cross-reactivity.The estab-lishment of this method provides a powerful tool for early diagnosis of ASF.

Radical resection of mid-and low-rectal cancer requires not only oncologic safety but also preservation of organs and postoperative bowel function.While a 1-2 cm distal resection margin has been largely accepted,the optimal length of the proximal margin remains highly controversial.Clinically,the"10-cm rule"derived from colon cancer is often referenced,yet its applicability to rectal cancer lacks consistent supporting evidence.Previous studies have shown that an excessively long proximal margin may increase anastomotic tension and lead to anastomotic leakage,whereas insufficient resection heightens the risk of positive margins and local recurrence.In addition,the extent of lymph node metastasis,vascular perfusion of the proximal bowel,radiation-induced injury after neoadjuvant chemoradiotherapy,and postoperative bowel function-particularly low anterior resection syndrome-are all important factors influencing the selection of the proximal margin.In recent years,the application of indocyanine green fluorescence imaging has provided new evidence for intraoperative assessment of bowel perfusion;for patients receiving neoadjuvant chemoradiotherapy,radiation injury presents a gradient pattern,and resecting approximately≥20 cm proximal to the tumor may reduce the incidence of anastomosis-related complications.Based on current literature,this review provides a systematic overview of the historical evolution,influencing factors,and clinical evidence regarding proximal resection margins in rectal cancer surgery,with the aim of informing individualized margin selection and optimizing surgical strategies.

BACKGROUND:Epidemiologic investigations and some experiments have shown that there is a close relationship between peripheral blood cells and osteoporosis,but the causal relationship between the two at the genetic level is still unclear. OBJECTIVE:To explore the causal relationship between peripheral blood cells and osteoporosis using Mendelian randomization methods. METHODS:Genome-wide association study data sets on peripheral blood cells,overall bone density at different ages,and calcaneal bone density were obtained from databases such as Blood Cell Consortium and MRC Integrative Epidemiology Unit. Blood cells were used as exposure data,with bone density at different ages and calcaneal bone density serving as outcome data. Mendelian randomization analyses were performed using methods such as inverse variance weighting,MR-Egger,weighted median method,and simple median. The results were assessed for heterogeneity,pleiotropy,and sensitivity using Cochran's Q,MR-Egger regression,and Leave-one-out method. The causal relationship between exposure and outcomes was evaluated using β values. RESULTS AND CONCLUSION:Due to the heterogeneity revealed by Cochran's Q test in the Mendelian randomization results,the results of the study were based on the inverse variance weighting method. The inverse variance weighting results showed that when age-specific bone density was used as an outcome,there was a negative causal relationship between white blood cell count and whole-body bone mineral density at the age of 45-60 years[β=-0.07,95％ confidence interval (CI):-0.13,-0.01,P=0.02],a positive causal relationship between monocyte count and whole-body bone mineral density at the age of 45-60 years (β=0.05,95％ CI:0.00,0.10,P=0.037),a negative causal relationship between white blood cell and basophil counts and whole-body bone mineral density over 60 years old (β=-0.04,95％ CI:-0.07,-0.01,P=0.005;β=-0.04,95％ CI:-0.07,-0.00,P=0.038),a positive causal relationship between hemoglobin concentration and hematocrit and whole-body bone mineral density over 60 years old (β=0.04,95％ CI:0.01,0.08,P=0.012;β=0.04,95％ CI:0.00,0.07,P=0.039),and a negative causal relationship between white cell count and whole-body bone mineral density at an undistinguished age (β=-0.10,95％ CI:-0.16,-0.03,P=0.002). When heel bone mineral density was used as an outcome,there was a negative causal relationship between white cell count and heel bone mineral density (β=-0.04,95％ CI:-0.07,-0.01,P=0.016),and a positive causal relationship between hemoglobin concentration and hematocrit and heel bone mineral density (β=0.05,95％ CI:0.01,0.08,P=0.007;β=0.05,95％ CI:0.01,0.08,P=0.004). To ensure the robustness of the results,meta-analyses of Mendelian randomization results of peripheral blood cells and whole-body bone mineral density as well as heel bone mineral density in different age groups were conducted. The results suggested that for every standard deviation decrease in log-transformed white blood cell count,there was a 5％ reduction in the risk of decreased bone mineral density (OR=0.95,95％ CI:0.94,0.97,P<0.001);whereas for every standard deviation increase in hemoglobin concentration and hematocrit,there was a 4％ reduction in the risk of decreased bone density (OR=1.04,95％ CI:1.03,1.06,P<0.001). In conclusion,increased white blood cell count in peripheral blood is a risk factor for bone mineral density;whereas increased hematocrit and hemoglobin concentration are protective factors for bone mineral density.

Objective:To investigate the effects of circular RNA(circRNA)hsa_circ_0081621 on the malignant biological behaviors of human laryngeal squamous cell carcinoma AMC-HN-8 and TU177 cells.Methods:AMC-HN-8 and TU177 cells were routinely cultured.si-NC,si-hsa_circ_0081621,empty vector(vector),and hsa_circ_0081621 overexpression vector(hsa_circ_0081621-OE)were transfected into AMC-HN-8 and TU177 cells,namely si-NC,si-hsa_circ_0081621,vector,and hsa_circ_0081621-OE groups,respectively.The effects of knockdown or overexpression of hsa_circ_0081621 on the proliferation,migration,and invasion of AMC-HN-8 and TU177 cells were detected by CCK-8 assay,colony formation assay,scratch wound healing assay,and Transwell chamber assay.Results:Successful knockdown or overexpression of hsa_circ_0081621 was achieved in AMC-HN-8 and TU177 cells.hsa_circ_0081621 knockdown significantly inhibited while hsa_circ_0081621 overexpression significantly promoted the proliferation,migration,and invasion of AMC-HN-8 and TU177 cells(P<0.01 or P<0.001 or P<0.0001).Conclusion:hsa_circ_0081621 promotes the malignant biological behaviors of human laryngeal squamous cell carcinoma AMC-HN-8 and TU177 cells.

African swine fever(ASF)is an acute,febrile,and highly fatal disease caused by African swine fever virus(ASFV)in pigs.Given the current lack of commercial vaccines and the continu-ous evolution of ASFV in recent years,the emergence of moderately virulent genotype Ⅱ strains and the introduction of genotype Ⅰ attenuated strains have led to persistent and chronic infections in pigs.Therefore,the detection of specific antibodies against ASFV has become imperative.In this study,we established a competitive chemiluminescence immunoassay(p30-cCLIA)for detecting ASFV p30 antibodies using p30 monoclonal antibodies.By detecting sera with clear negative and positive backgrounds,we determined that the Cut-off value of this method was 50％,with both di-agnostic sensitivity(Dsn)and diagnostic specificity(Dsp)reaching 100％.Under optimal reaction conditions,we screened out an enzyme-labeled stabilizer suitable for p30 monoclonal antibody 16-5E7E8-HRP.Furthermore,the sensitivity of the established p30-cCLIA method was higher than that of the commercial blocking ELISA kit(1∶2 048 vs 1∶512)and exhibited good repeatability.Detection of sera positive for other porcine virus infections showed no cross-reactivity.The estab-lishment of this method provides a powerful tool for early diagnosis of ASF.

BACKGROUND:Epidemiologic investigations and some experiments have shown that there is a close relationship between peripheral blood cells and osteoporosis,but the causal relationship between the two at the genetic level is still unclear. OBJECTIVE:To explore the causal relationship between peripheral blood cells and osteoporosis using Mendelian randomization methods. METHODS:Genome-wide association study data sets on peripheral blood cells,overall bone density at different ages,and calcaneal bone density were obtained from databases such as Blood Cell Consortium and MRC Integrative Epidemiology Unit. Blood cells were used as exposure data,with bone density at different ages and calcaneal bone density serving as outcome data. Mendelian randomization analyses were performed using methods such as inverse variance weighting,MR-Egger,weighted median method,and simple median. The results were assessed for heterogeneity,pleiotropy,and sensitivity using Cochran's Q,MR-Egger regression,and Leave-one-out method. The causal relationship between exposure and outcomes was evaluated using β values. RESULTS AND CONCLUSION:Due to the heterogeneity revealed by Cochran's Q test in the Mendelian randomization results,the results of the study were based on the inverse variance weighting method. The inverse variance weighting results showed that when age-specific bone density was used as an outcome,there was a negative causal relationship between white blood cell count and whole-body bone mineral density at the age of 45-60 years[β=-0.07,95％ confidence interval (CI):-0.13,-0.01,P=0.02],a positive causal relationship between monocyte count and whole-body bone mineral density at the age of 45-60 years (β=0.05,95％ CI:0.00,0.10,P=0.037),a negative causal relationship between white blood cell and basophil counts and whole-body bone mineral density over 60 years old (β=-0.04,95％ CI:-0.07,-0.01,P=0.005;β=-0.04,95％ CI:-0.07,-0.00,P=0.038),a positive causal relationship between hemoglobin concentration and hematocrit and whole-body bone mineral density over 60 years old (β=0.04,95％ CI:0.01,0.08,P=0.012;β=0.04,95％ CI:0.00,0.07,P=0.039),and a negative causal relationship between white cell count and whole-body bone mineral density at an undistinguished age (β=-0.10,95％ CI:-0.16,-0.03,P=0.002). When heel bone mineral density was used as an outcome,there was a negative causal relationship between white cell count and heel bone mineral density (β=-0.04,95％ CI:-0.07,-0.01,P=0.016),and a positive causal relationship between hemoglobin concentration and hematocrit and heel bone mineral density (β=0.05,95％ CI:0.01,0.08,P=0.007;β=0.05,95％ CI:0.01,0.08,P=0.004). To ensure the robustness of the results,meta-analyses of Mendelian randomization results of peripheral blood cells and whole-body bone mineral density as well as heel bone mineral density in different age groups were conducted. The results suggested that for every standard deviation decrease in log-transformed white blood cell count,there was a 5％ reduction in the risk of decreased bone mineral density (OR=0.95,95％ CI:0.94,0.97,P<0.001);whereas for every standard deviation increase in hemoglobin concentration and hematocrit,there was a 4％ reduction in the risk of decreased bone density (OR=1.04,95％ CI:1.03,1.06,P<0.001). In conclusion,increased white blood cell count in peripheral blood is a risk factor for bone mineral density;whereas increased hematocrit and hemoglobin concentration are protective factors for bone mineral density.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.