AIM To investigate the protective effects of Shuangyi Qushi Tongluo Capsules(Shuangyi Capsules)on Dexamethasone(Dex)induced osteoporosis in mice.METHODS The C57BL/6J mice were randomly divided into the control group,the model group,the Xianling Gubao Capsules group(1.5 g/kg),and the low-dose,moderate-dose,and high-dose Shuangyi Capsules groups(0.6,1.2,and 2.4 g/kg).The mouse model of osteoporosis was induced by 8-week intraperitoneal injection of Dex sodium phosphate injection(5 mg/kg).The mice had their femur osteogenesis observed with hematoxylin and eosin(HE)staining and tartrate-resistant acid phosphatase(TRAP)staining;their serum alkaline phosphatase(ALP)and osteocalcin(BGP)activities detected by ELISA;their femoral mRNA expressions of Col-Ⅰ,OCN,and OPN detected by RT-qPCR;and their femoral protein expressions of OPG and RANKL detected by Western blot.Upon the MC3T3-E1 cells exposed to Dex and Shuangyi Capsules,their viability was evaluated by CCK-8 assay;their mineralization determined by alkaline phosphatase staining and alizarin red staining(ARS);and their intracellular ROS level detected using DCFH-DA probe.RESULTS Compared with the model group,Shuangyi Capsules groups demonstrated improved fracture of femoral trabeculae and reduced number of osteoclasts;increased serum ALP and BGP activities(P＜0.05,P＜0.01);increased femoral expressions of Col-Ⅰ mRNA and OPG protein(P＜0.05,P＜0.01);and decreased RANKL protein expression(P＜0.05).Compared with the MC3T3-E1 cells stimulated by Dex,those underwent further treatment of Shuangyi Capsules demonstrated increased cell viability and ALP activity(P＜0.05,P＜0.01);increased mineralization and calcium nodule formation;increased expressions of Col-Ⅰ,OCN,OPN mRNA and OPG protein(P＜0.05,P＜0.01);decreased RANKL protein expression(P＜0.05,P＜0.01);and reduced ROS levels.CONCLUSION Shuangyi Capsules ameliorate Dex-induced osteoporosis in mice by suppressing osteoclast overactivation,enhancing osteoblast activity,and stimulating bone formation through modulation of Col-Ⅰ,OCN,OPN mRNA and OPG/RANKL protein levels.

Objective To elucidate the impact of chronic kidney disease(CKD)on the clinical outcomes of patients with left main coronary artery disease(LMCAD)undergoing intravascular ultrasound(IVUS)-guided percutaneous coronary intervention(PCI).Methods This retrospective study enrolled consecutive patients with LMCAD who underwent IVUS-guided PCI at Wuhan Asia Heart Hospital between January 2017 and December 2020.Patients were stratified into CKD and non-CKD groups according to the presence of CKD.Clinical data were systematically retrieved from the electronic health record system.Demographic,clinical,and angiographic characteristics were compared between groups.The primary endpoint was major adverse cardiovascular events(MACE),defined as a composite of all-cause mortality,myocardial infarction,and ischemic stroke.Results A total of 325 LMCAD patients[mean age(62.56±9.86)years;73.54％male]were included,with 31 patients(9.54％)in the CKD group.During a median follow-up of 5 years,CKD patients exhibited significantly older age[(70.13±9.77)years vs.(61.77±9.54)years,P＜0.001],higher prevalence of three-vessel disease(64.52％vs.38.10％;P=0.040)and left main bifurcation lesion(45.16％vs.37.76％,P=0.011),greater IVUS-detected calcification burden(P=0.029),and higher median SYNTAXⅡ scores[(34.10(30.30,39.25)vs.26.75(22.42,31.58),P＜0.001)].The cumulative incidence of MACE was significantly higher in the CKD group compared to the non-CKD group(32.26％vs.9.18％,P＜0.001).Univariate Cox regression analysis and Kaplan-Meier survival curves confirmed a 5.877-fold increased risk of MACE in CKD patients(95％CI 2.765-12.494).After adjusting for age and cardiac function,CKD remained an independent predictor of MACE(HR 3.611,95％CI 1.634-7.978).Conclusions LMCAD patients with concomitant CKD present with advanced age,impaired cardiac function,more extensive coronary disease,and severe calcification.The presence of CKD is associated with a significantly worse long-term prognosis.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

Objective To elucidate the impact of chronic kidney disease(CKD)on the clinical outcomes of patients with left main coronary artery disease(LMCAD)undergoing intravascular ultrasound(IVUS)-guided percutaneous coronary intervention(PCI).Methods This retrospective study enrolled consecutive patients with LMCAD who underwent IVUS-guided PCI at Wuhan Asia Heart Hospital between January 2017 and December 2020.Patients were stratified into CKD and non-CKD groups according to the presence of CKD.Clinical data were systematically retrieved from the electronic health record system.Demographic,clinical,and angiographic characteristics were compared between groups.The primary endpoint was major adverse cardiovascular events(MACE),defined as a composite of all-cause mortality,myocardial infarction,and ischemic stroke.Results A total of 325 LMCAD patients[mean age(62.56±9.86)years;73.54％male]were included,with 31 patients(9.54％)in the CKD group.During a median follow-up of 5 years,CKD patients exhibited significantly older age[(70.13±9.77)years vs.(61.77±9.54)years,P＜0.001],higher prevalence of three-vessel disease(64.52％vs.38.10％;P=0.040)and left main bifurcation lesion(45.16％vs.37.76％,P=0.011),greater IVUS-detected calcification burden(P=0.029),and higher median SYNTAXⅡ scores[(34.10(30.30,39.25)vs.26.75(22.42,31.58),P＜0.001)].The cumulative incidence of MACE was significantly higher in the CKD group compared to the non-CKD group(32.26％vs.9.18％,P＜0.001).Univariate Cox regression analysis and Kaplan-Meier survival curves confirmed a 5.877-fold increased risk of MACE in CKD patients(95％CI 2.765-12.494).After adjusting for age and cardiac function,CKD remained an independent predictor of MACE(HR 3.611,95％CI 1.634-7.978).Conclusions LMCAD patients with concomitant CKD present with advanced age,impaired cardiac function,more extensive coronary disease,and severe calcification.The presence of CKD is associated with a significantly worse long-term prognosis.

AIM To investigate the protective effects of Shuangyi Qushi Tongluo Capsules(Shuangyi Capsules)on Dexamethasone(Dex)induced osteoporosis in mice.METHODS The C57BL/6J mice were randomly divided into the control group,the model group,the Xianling Gubao Capsules group(1.5 g/kg),and the low-dose,moderate-dose,and high-dose Shuangyi Capsules groups(0.6,1.2,and 2.4 g/kg).The mouse model of osteoporosis was induced by 8-week intraperitoneal injection of Dex sodium phosphate injection(5 mg/kg).The mice had their femur osteogenesis observed with hematoxylin and eosin(HE)staining and tartrate-resistant acid phosphatase(TRAP)staining;their serum alkaline phosphatase(ALP)and osteocalcin(BGP)activities detected by ELISA;their femoral mRNA expressions of Col-Ⅰ,OCN,and OPN detected by RT-qPCR;and their femoral protein expressions of OPG and RANKL detected by Western blot.Upon the MC3T3-E1 cells exposed to Dex and Shuangyi Capsules,their viability was evaluated by CCK-8 assay;their mineralization determined by alkaline phosphatase staining and alizarin red staining(ARS);and their intracellular ROS level detected using DCFH-DA probe.RESULTS Compared with the model group,Shuangyi Capsules groups demonstrated improved fracture of femoral trabeculae and reduced number of osteoclasts;increased serum ALP and BGP activities(P＜0.05,P＜0.01);increased femoral expressions of Col-Ⅰ mRNA and OPG protein(P＜0.05,P＜0.01);and decreased RANKL protein expression(P＜0.05).Compared with the MC3T3-E1 cells stimulated by Dex,those underwent further treatment of Shuangyi Capsules demonstrated increased cell viability and ALP activity(P＜0.05,P＜0.01);increased mineralization and calcium nodule formation;increased expressions of Col-Ⅰ,OCN,OPN mRNA and OPG protein(P＜0.05,P＜0.01);decreased RANKL protein expression(P＜0.05,P＜0.01);and reduced ROS levels.CONCLUSION Shuangyi Capsules ameliorate Dex-induced osteoporosis in mice by suppressing osteoclast overactivation,enhancing osteoblast activity,and stimulating bone formation through modulation of Col-Ⅰ,OCN,OPN mRNA and OPG/RANKL protein levels.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

ObjectiveRapid and accurate identification of body fluid traces at crime scenes is crucial for case investigation. Leveraging the speed and sensitivity of nucleic acid detection technology based on SHERLOCK, our research focuses on developing a peripheral blood SHERLOCK-HBA detection system to detect mRNA in forensic practice. MethodsShort crRNA fragments targeting the blood-specific mRNA gene HBA were designed and screened, alongside RPA primers. Optimal RPA primers were selected based on specificity and amplification efficiency, leading to the establishment of the RPA system. The most efficient crRNA was chosen based on relative fluorescence units (RFU) generated by the Cas protein reaction, and the Cas protein reaction system was constructed to establish the SHERLOCK-HBA detection method. The RPA and Cas protein reaction systems in the SHERLOCK detection system were then individually optimized. A total of 79 samples of five body fluids were tested to evaluate the method’s ability to identify blood, with further verification through species-specific tests, sensitivity tests, mixed spots detection, aged samples, UV-irradiated samples, and actual casework samples. ResultsThe SHERLOCK reaction system for the peripheral blood-specific marker HBA was successfully established and optimized, enabling detection within 30 min. The method demonstrated a detection limit of 0.001 ng total RNA, better than FOB strip method and comparable to RT-PCR capillary electrophoresis. The system could detect target body fluids in mixed samples and identify blood in samples stored at room temperature for three years and exposed to UV radiation for 32 h. Detection of 11 casework samples showed performance comparable to RT-PCR capillary electrophoresis. ConclusionThis study presents a CRISPR/Cas-based SHERLOCK-HBA detection system capable of accurately, sensitively, and rapidly identifying blood samples. Introducing CRISPR/Cas technology to forensic body fluid identification represents a significant advancement in applying cutting-edge molecular biology techniques to forensic science.The method’s simplicity, shorter detection time, and independence from specialized equipment make it promising for rapid blood sample identification in forensic cases.

Objective To investigate the effect of overexpression of LncRNA MEG3 on proliferation,migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism.Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma(HCC)was analyzed by online bioinformatics analysis,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MEG3 expression in different HCC cell lines.A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells,and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay.CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells.A reactive oxygen species(ROS)fluorescence probe(DCFH-DA)and malondialdehyde(MDA)kit were used to assess the changes in ROS production and MDA level in the cells.Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells,which was consistent with the results of bioinformatic analysis(P<0.05).Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration(P<0.05),increased the cell inhibition rate of cisplatin(P<0.05),enhanced cellular ROS production and increased MDA levels in the cells(P<0.05).MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.Conclusion The expression of MEG3 is decreased in HCC cells,and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

Objective To investigate the effect of overexpression of LncRNA MEG3 on proliferation,migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism.Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma(HCC)was analyzed by online bioinformatics analysis,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MEG3 expression in different HCC cell lines.A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells,and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay.CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells.A reactive oxygen species(ROS)fluorescence probe(DCFH-DA)and malondialdehyde(MDA)kit were used to assess the changes in ROS production and MDA level in the cells.Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells,which was consistent with the results of bioinformatic analysis(P<0.05).Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration(P<0.05),increased the cell inhibition rate of cisplatin(P<0.05),enhanced cellular ROS production and increased MDA levels in the cells(P<0.05).MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.Conclusion The expression of MEG3 is decreased in HCC cells,and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

Objective To compare the clinical curative effect of retroauricular and tympanic injection of methylprednisolone sodium succinate combined with hyperbaric oxygen in nerve deafness.Methods A retrospective analysis was performed on the case data with nerve deafness.According to cohort method,they were divided into retroauricular injection group(retroauricular injection of 40 mg methylprednisolone sodium succinate+hyperbaric oxygen therapy)and tympanic injection group(tympanic injection of 40 mg methylprednisolone sodium succinate+hyperbaric oxygen therapy).Both groups were treated for 7 times(once every other day).The curative effect,pure tone hearing threshold,oxidative stress and inflammatory response indexes were compared between the two groups,and adverse reactions were statistically analyzed.Results There were 56 cases in retroauricular injection group and 60 cases in tympanic injection group.There was no significant difference in total response rate between retroauricular injection group and tympanic injection group(92.86％vs 88.33％,P＞0.05).After treatment,average pure tone hearing thresholds in retroauricular injection group and tympanic injection group were(38.49±5.71)and(40.35±6.23)dB;high-frequency hearing thresholds were(61.36±6.52)and(63.42±7.09)dB;low-frequency hearing thresholds were(59.72±6.85)and(60.81±7.26)dB;levels of superoxide dismutase(SOD)were(112.38±10.72)and(110.43±10.29)U·mL-1;levels of lipid peroxide(LPO)were(4.69±1.08)and(5.03±1.12)nmol·mL-1;endothelin(ET)levels were(2.11±0.76)and(2.34±0.81)ng·L-1;interleukin-6(IL-6)levels were(31.79±4.27)and(33.18±4.98)pg·mL-1;levels of C-reactive protein(CRP)were(5.25±1.48)and(5.72±1.62)mg·L-1;levels of tumor necrosis factor-α(TNF-α)were(45.33±4.61)and(46.95±5.24)ng·L-;the difference was not statistically significant(all P＞0.05).During treatment,there were significant differences in total incidence of adverse reactions between retroauricular injection group and tympanic injection group(23.21％vs 51.67％,P＜0.05).There were 11 case times of local pain and 4 case times of vertigo in retroauricular injection group,while there were 30 case times of local pain and 15 case times of vertigo in tympanic injection group.Conclusion Retroauricular injection or tympanic injection of methylprednisolone sodium succinate combined with hyperbaric oxygen therapy has the comparable curative effect in patients with nerve deafness;the two administration modes both can promote hearing recovery,relieve reduce oxidative stress and inflammatory response.However,retroauricular injection has lower incidence of local pain and vertigo.