ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease （AD） through regulating mitochondrial Ca²⁺ homeostasis and kinetic balance based on the protein kinase A （PKA）/calcineurin （CaN） signaling pathway. MethodsSixty three-month-old amyloid precursor protein （APP）/presenilin 1 （PS1） double transgenic mice were randomly divided into a model group， a donepezil group（0.65 mg·kg^-1）， a low-dose Yizhi Qingxin prescription group （YQF-L，2.6 g·kg^-1）， a medium-dose Yizhi Qingxin prescription group （YQF-M，5.2 g·kg^-1）， and a high-dose Yizhi Qingxin prescription group （YQF-H，10.4 g·kg^-1）， with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily， with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition （NOR） and Morris water maze （MWM） tests. Hematoxylin-eosin （HE）/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy （TEM） was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester （Fluo-4 AM） Ca²⁺ probe was used to measure intracellular Ca²⁺ concentration in brain tissue. Western blot was used to determine the protein expression of PKA， CaN， sodium/calcium/lithium exchanger （NCLX）， mitochondrial calcium uniporter （MCU）， calmodulin （CaM）， dynamin-related protein 1 （Drp1）， and phosphorylated dynamin-related protein 1 （serine 637 site） ［p-Drp1（S637）］ in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to measure the expression of PKA， CaN， CaM， NCLX， MCU， and Drp1 mRNAs. ResultsCompared with those in the normal group， the recognition index （RI） of the model group decreased （P0.01）， and the number of crossings through the original platform area， the duration of stay in the target quadrant， and the distance were reduced （P0.01）. The protein expression of PKA， NCLX， and p-DRP1 （ser637） significantly decreased （P0.05）， and the mRNA expression of PKA and NCLX significantly decreased （P0.05）. The escape latency （EL） was prolonged （P0.05）， and the intracellular Ca²⁺ level significantly increased （P0.01）. The protein expression of CaN， CaM， MCU， and Drp1， as well as the mRNA expression of CaN， MCU， and Drp1， significantly increased （P0.05）. After intervention with Donepezil and Yizhi Qingxin prescription， compared with that in the model group， the RI of the treatment group significantly increased （P0.05）， and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased （P0.05）. The protein expression of PKA， NCLX， and p-Drp1 （ser637） and the mRNA expression of PKA and NCLX significantly increased （P0.05）. On the 4th and 5th days， the EL was shortened （P0.05）， and the intracellular Ca²⁺ level decreased （P0.05）. The protein expression of CaN， CaM， MCU， and Drp1 and the mRNA expression of CaN， MCU， and Drp1 significantly decreased （P0.05）. ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway， upregulates the expression of PKA， NCLX， and p-Drp1 （ser637） proteins， reduces the expression of CaN， CaM， MCU， and Drp1 proteins， and regulates Ca²⁺ homeostasis and mitochondrial dynamic balance， thereby enhancing the spatial learning and memory abilities of AD mice.

ObjectiveTo explore the mechanism by which Yizhi Qingxin prescription improves mitochondrial dysfunction in Alzheimer's disease （AD） through regulating mitochondrial Ca²⁺ homeostasis and kinetic balance based on the protein kinase A （PKA）/calcineurin （CaN） signaling pathway. MethodsSixty three-month-old amyloid precursor protein （APP）/presenilin 1 （PS1） double transgenic mice were randomly divided into a model group， a donepezil group（0.65 mg·kg^-1）， a low-dose Yizhi Qingxin prescription group （YQF-L，2.6 g·kg^-1）， a medium-dose Yizhi Qingxin prescription group （YQF-M，5.2 g·kg^-1）， and a high-dose Yizhi Qingxin prescription group （YQF-H，10.4 g·kg^-1）， with 12 mice in each group. Twelve C57BL/6J mice with the same genetic background served as a normal group. Each treatment group received gavage administration daily， with the model and normal groups receiving equal volume of physiological saline. Intervention continued for 12 consecutive weeks. The learning and memory abilities of the mice were assessed using the novel object recognition （NOR） and Morris water maze （MWM） tests. Hematoxylin-eosin （HE）/Nissl staining was used to observe histopathological changes in the hippocampus. Transmission electron microscopy （TEM） was used to observe mitochondrial ultrastructure. Fluo-4 acetoxymethyl ester （Fluo-4 AM） Ca²⁺ probe was used to measure intracellular Ca²⁺ concentration in brain tissue. Western blot was used to determine the protein expression of PKA， CaN， sodium/calcium/lithium exchanger （NCLX）， mitochondrial calcium uniporter （MCU）， calmodulin （CaM）， dynamin-related protein 1 （Drp1）， and phosphorylated dynamin-related protein 1 （serine 637 site） ［p-Drp1（S637）］ in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to measure the expression of PKA， CaN， CaM， NCLX， MCU， and Drp1 mRNAs. ResultsCompared with those in the normal group， the recognition index （RI） of the model group decreased （P0.01）， and the number of crossings through the original platform area， the duration of stay in the target quadrant， and the distance were reduced （P0.01）. The protein expression of PKA， NCLX， and p-DRP1 （ser637） significantly decreased （P0.05）， and the mRNA expression of PKA and NCLX significantly decreased （P0.05）. The escape latency （EL） was prolonged （P0.05）， and the intracellular Ca²⁺ level significantly increased （P0.01）. The protein expression of CaN， CaM， MCU， and Drp1， as well as the mRNA expression of CaN， MCU， and Drp1， significantly increased （P0.05）. After intervention with Donepezil and Yizhi Qingxin prescription， compared with that in the model group， the RI of the treatment group significantly increased （P0.05）， and the number of crossings through the platform and the duration of stay in the target quadrant significantly increased （P0.05）. The protein expression of PKA， NCLX， and p-Drp1 （ser637） and the mRNA expression of PKA and NCLX significantly increased （P0.05）. On the 4th and 5th days， the EL was shortened （P0.05）， and the intracellular Ca²⁺ level decreased （P0.05）. The protein expression of CaN， CaM， MCU， and Drp1 and the mRNA expression of CaN， MCU， and Drp1 significantly decreased （P0.05）. ConclusionYizhi Qingxin prescription regulates the PKA/CaN pathway， upregulates the expression of PKA， NCLX， and p-Drp1 （ser637） proteins， reduces the expression of CaN， CaM， MCU， and Drp1 proteins， and regulates Ca²⁺ homeostasis and mitochondrial dynamic balance， thereby enhancing the spatial learning and memory abilities of AD mice.

Objective: To investigate the characteristics and potential risk factors associated with HIV infection among newly reported HIV positive male student cases in Jiangsu Province from 2023 to 2024, so as to provide evidence for targeted intervention strategies. Methods: Data were obtained from the China CDC Surveillance System on newly reported HIV positive male student cases from 2023 to 2024. A survey was conducted to collect information on demographic characteristics, knowledge of AIDS prevention, education and training history, HIV testing history, behavioral and substance use patterns, and other relevant factors prior to HIV diagnosis among 343 newly reported HIV positive male student cases in Jiangsu Province. Multivariate Logistic regression analysis was used to assess the risk factors of HIV infection among male students. Results: Among the HIV positive male student cases, homosexual behavior accounted for 93.88% of transmission routes, while 10.20% involved heterosexual contact, as well as 4.08% for two sexual hehaviors. Awareness of HIV prevention knowledge was 97.08%, and 66.76% had previously undergone HIV testing. Among the respondents, 10.50% had used rush poppers as enhancers, and 72.30% had received HIV prevention education within the past year. Among students cases with homosexual behavior, the median time from first homosexual contact to HIV diagnosis was ≤2 years, with 54.66% of cases falling into this category; the most common way of finding same sex partners was through social software, accounting for 88.20% of cases, while the proportion of those via "Blued" app reached 87.07% ; the proportion of using condoms every time during sexual activity in the past six months was 12.27%. Among the student cases with homosexual behavior, the results of multiple Logistic regression analysis showed that student cases aged 18 to 24 ( OR =4.52) and >24 ( OR = 19.23 ), without receiving education on HIV prevention in the past year ( OR =1.86), having consistent condom use ( OR =2.73) and not using condoms ( OR =2.12) during the last sexual activity were more likely to had the first same sex sexual activity for more than 3 years before being diagnosed as HIV positive cases (all P <0.05). Student cases who were uncertain about their partner s sexual identity ( OR =0.33), and who primarily identified same sex partners through "other" means ( OR = 0.23 ) were more likely to avoid HIV testing; in contrast, student cases with consistent condom use during the last homosexual encounter ( OR =7.20) was significantly associated with increased likelihood of HIV testing (all P <0.05). Conclusions Newly reported HIV positive male student cases in Jiangsu Province exhibit serious discrepancies between knowledge and practice regarding HIV prevention. Measures are needed to accelerate the optimization of campus based HIV prevention education content and delivery methods. Simultaneously, enhanced management of extracurricular male populations is essential to effectively control the spread of HIV.

OBJECTIVE: To analyze the time characteristics of the Ethos online adaptive radiotherapy (OART) process in clinical practice and provide guidance for the comprehensive optimization of each stage of adaptive radiotherapy. METHODS: The study involved 61 patients with cervical, rectal, gastric, lung, esophageal, and breast cancers who underwent Ethos OART. The mean ± standard deviation of segmental time, total time, and target volume for these patients were tracked. The time characteristics for different cancer types were evaluated, and the average time for target and organ at risk (OAR) modifications was compared with the average target volume for each cancer type. RESULTS: Cervical cancer born the longest total treatment time, while breast cancer had the shortest. For all cancer types except breast cancer, the modification time for target and OAR was the most time-consuming segment. The average time for target and OAR modifications aligned with the trend of the average target volume. CONCLUSION The total treatment time for various cancers ranges from 15 to 35 minutes, indicating room for improvement.
Humans ; Radiotherapy Planning, Computer-Assisted/methods* ; Neoplasms/radiotherapy* ; Female

Objective:To discuss the effect of oridonin on the proliferation,migration,epithelial-mesenchymal transition(EMT),and apoptosis of the human nasopharyngeal carcinoma HONE-1 cells,and to clarify its related antitumor mechanism.Methods:The HONE-1 cells were treated with different concentrations(0,5,10,20,40,80,and 160 mg·L-1)of oridonin for 48 h.CCK-8 method was used to detect the inhibitory rates of proliferation of the cells in various groups and the drug concentration for subsequent experiment was confirmed.The HONE-1 cells were divided into control group,3 mg·L-1 oridonin group,and 6 mg·L-1 oridonin group.After 24 and 48 h of culture,CCK-8 method was used to detect the proliferation activities of the cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)method was used to detect the rates of EdU-positive cells in various groups;colony formation assay was used to detect the numbers of clone formation in the cells in various groups;Transwell chamber experiment and cell wound assay were used to detect the numbers of migration cells and the scratch healing rates of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of cyclin-dependent kinase 1(CDK1)and cyclin-dependent kinase 4(CDK4)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of E-cadherin,Vimentin,Caspase-3,and poly ADP-ribose polymerase 1(PARP1)proteins in the cells in various groups.Results:The CCK-8 method results showed that the half-maximal inhibitory concentration(IC50)of oridonin at 48 h was 12.18 mg·L-1,and 1/4 IC50 and 1/2 IC50 values were used as the concentrations for subsequent experiments.Compared with control group,after treated for 24 and 48 h,the proliferation activities of the cells in 3 and 6 mg·L-1 oridonin groups were decreased(P＜0.05 or P＜0.01),the rate of EdU-positive cells were decreased(P＜0.05 or P＜0.01),the numbers of clone formation and migraton cells were decreased(P＜0.05 or P＜0.01),the scratch healing rates were decreased(P＜0.05 or P＜0.01),the expression levels of CDK1 and CDK4 mRNA in the cells were decreased(P＜0.05 or P＜0.01),the expression levels of E-cadherin,Caspase-3,and PARP1 proteins were increased(P＜0.05 or P＜0.01),and the expression levels of Vimentin protein were decreased(P＜0.05).Conclusion:Oridonin can inhibit the proliferation,clone formation,and migration of the human nasopharyngeal carcinoma HONE-1 cells by downregulating the expression of cell cycle-related proteins and EMT,and promote the apoptosis to exert an antitumor effect.

Ischemic cerebrovascular disease(ICVD)refers to the degeneration,necrosis,or transient loss of function of local brain tissue due to blood supply disorders,and is one of the common neurological diseases.Exosomes are a subgroup of extracellular vesicles,characterized by good biocompatibility,low toxicity and immunogenicity,and strong designability.The components of exosomes include proteins,lipids,genetic materials,etc.They come from a wide range of sources and can be secreted by various cells under physiological or pathological conditions.The roles and related mechanisms of exosomes from different sources in the treatment of ICVD are different.This article reviews the therapeutic effects and related mechanisms of exesomes derived from mesenchymal stem cells,brain cells,blood,endothelial cells,and macrophages on ICVD,in order to provide reference for the clinical treatment of ICVD.

Objective To observe the distribution and characteristics of lymphatic vessels in normal and injured mouse spinal cord,and to determine if lymphatic vessels participate in the repair of spinal cord injury.Methods Thirty-nine adult male KM mice were divided randomly into a normal group(n=6)and an injured group(n=33).Mice in the injured group were further divided randomly into mice examined on postoperative days 1,3,5,7,and 14.Three mice in the injury group died after acupuncture injury,and the sample was subsequently supplemented randomly.Spinal cord damage was induced in the injured group by acupuncture,while mice in the normal group had no spinal cord damage.The distribution of lymphatic endothelial cells(LECs)in the spinal cord was detected by immunohistochemistry,and expression levels of the lymphatic endothelial cell markers prospero-related homobox-1(prox-1),lymphatic vessel endothelial cell hyaluronic acid receptor-1(lyve-1),and flat foot protein(podoplanin),and the vascular endothelial cell marker CD34 were observed in the spinal cords in normal and acupuncture-injured mice.Spinal cord samples were examined by immunofluorescence staining,and the source of new LECs was explored by observing the co-expression of lyve-1/prox-1,lyve-1/podoplanin,and CD34/prox-1.Results Lymphangioid structures were present in the spinal cord in normal mice and were distributed in segments,laterally between the white matter and gray matter.Nascent lymphangioid-like structures appeared in the spinal cord at the site of acupuncture injury,and prox-1,podoplanin,lyve-1,and CD34 were expressed simultaneously;however,these nascent lymphangioid-like structures disappeared after scarring during spinal cord injury.Conclusions Segmental,transversely distributed lymphangioid-like structures are present in the spinal cord in normal adult mice,and neonatal lymphangioid-like structures are involved in the reconstruction of spinal cord injury.These nascent LECs may originate from the surrounding existing lymphatic vessels or from vascular endothelial cells.

Objective To explore the safety and feasibility of intravenous dexmedetomidine(Dex)combined with targeted infusion of remifentanil in endoscopic retrograde cholangiopancreatography(ERCP)anesthesia in older adult patients.Methods From January to August 2021,98 older adult patients(≥65 years old)undergoing ERCP were randomly divided into TRP and TRD groups.The TRP group was anesthetized with target-controlled infusion of propofol and remifentanil and the TRD group was treated with Dex combined with target-controlled infusion of remifentanil.mean arterial pressure(MAP),heart rate(HR),electrocardiogram(ECG),respiratory rate(RR),pulse oxygen saturation(SpO2),bispectral index(BIS)before anesthesia induction(T0),immediately after induction of anesthesia(T1),endoscopic introduction(T2),duodenal papilla intubation(T3),endoscopy withdrawal(T4)and postoperative awakening(T5)were observed.Arterial blood gas analysis at different time points(T0,every 15 min after anesthesia induction and T5),PaO2,and PaCO2,were recorded at the above mentioned time points;and the remifentanil concentration in target-controlled infusion,operation time,recovery time(from infusion of remifentanil to consciousness recovery),anesthesia recovery time(from consciousness recovery to leaving the operating room),intraoperative body movement,Aldrete scores out of the room,Visual Analogue Scale(VAS)at 60 min after surgery,occurrence of post-operative adverse reactions,as well as the satisfaction of anesthesiologists,endoscopists,and patients were recorded.Results Compared with the TRP group,MAP at T1 and T3,SpO2 and RR at T1,T2,T3,and T4,and BIS at T2,T3,T4,and T5 increased,whereas HR at T1,T2,T3,and T4 decreased;the number of mandibular rests,incidence of hypoxemia,Aldrete score,and satisfaction increased,whereas the VAS score at 60 min after surgery decreased in the TRD group(P<0.05).There were no statistically significant differences in postoperative adverse reactions,PaO2 and PaCO2,target-controlled infusion remifentanil concentration,operation time,recovery time,and anesthesia recovery time between the two groups.Conclusion Compared with the target-controlled infusion of propofol-remifentanil,intravenous infusion of Dex combined with target-controlled infusion of remifentanil can reduce the incidence of hypoxemia in older adult patients during ERCP surgery,and the anesthesia regimen can meet the anesthesia needs of ERCP surgery,which is safe and feasible.

Objective:To investigate the predictive ability of Glypican-3 (GPC3) positive hepatocellular carcinoma based on the hepatobiliary specific contrast agent gadoxetate disodium enhancement of the liver imaging reporting and data system version 2018 (LI-RADS v2018) imaging features, and to assess the relevant clinical imaging features for postoperative recurrence in GPC3 positive HCC patients.Methods:This study was a cohort study. A total of 122 hepatocellular carcinoma patients who underwent gadoxetate disodium enhanced MRI examination with hepatic tumor resection in Henan Provincial People′s Hospital from January 2017 to December 2021 were retrospectively collected, including 96 GPC3 positive and 26 GPC3 negative patients. The imaging features defined by LI-RADS v2018 of HCC lesions were analyzed. Patients were followed up for 40 months to determine recurrence free survival (RFS). The logistic regression was used to analyze the risk factors of GPC3 positivity. An imaging model, and a clinical-imaging model which combined the patient′s alpha-fetoprotein levels were constructed. The efficacy of the model for predicting GPC3 positivity was assessed using receiver operating characteristic curves. Kaplan-Meier method was used to draw the survival curve, and the log-rank test was used to compare the RFS between GPC3 positive and negative patients. Risk factors affecting the recurrence of GPC3 positive HCC were assessed by Cox regression.Results:The results of logistic multivariate regression analysis confirmed that rim enhancement ( OR=5.685, 95% CI 1.229-26.287, P=0.026) and irregular tumor margin at hepatobiliary phase ( OR=4.431, 95% CI 1.684-11.663, P=0.003) were independent risk factors for GPC3 positive HCC. The area under the curve for predicting GPC3 positivity was 0.745 (95% CI 0.636-0.854) for the imaging model and 0.776 (95% CI 0.677-0.876) for the clinical-imaging model. The mean RFS in the GPC3 positive group was 22 months, and it was 32 months in the negative group. There was a statistically significant difference in RFS between the two groups ( χ2=5.15, P=0.023). The multivariate Cox regression analysis showed that the arterial rim enhancement ( HR=5.460, 95% CI 1.966-15.162, P=0.001), microvascular invasion ( HR=2.402, 95% CI 1.210-4.769, P=0.012), portal vein tumor thrombus ( HR=3.226, 95% CI 1.114-9.344, P=0.031) were independent risk factors for recurrence after hepatic tumor resection for GPC3-positive HCC. Conclusions:A model based on the LI-RADS v2018 imaging features of hepatobiliary specific contrast agent gadoxetate disodium enhancement can effectively predict GPC3 positive HCC. The arterial rim enhancement, microvascular invasion and portal vein tumor thrombus are independent risk factors for postoperative recurrence of GPC3 positive HCC.

Background and objective Pembrolizumab(PEM)has been shown to be effective in clinical trials for the treatment of advanced non-small cell lung cancer(NSCLC),but clinical trials were based on cohorts of patients selected on specific criteria,and whether the findings are consistent with real-world patients is debatable.The aim of this study is to evaluate the efficacy and safety of PEM in the treatment of advanced NSCLC based on real-world data.Methods A retro-spective collection of real-world data from patients with advanced NSCLC receiving PEM was conducted.Propensity score matching was used to eliminate inter-group differences and assess the efficacy and safety of PEM compared to chemotherapy.Results Among 450 matched patients,the incidence rates of any-grade adverse events were 79.87％in the PEM group and86.71％inthe chemotherapy group,while the incidence rates of grade＞3 adverse events were 4.03％and 7.31％,respectively.The objective response rates were 48.63％for PEM and 36.00％for chemotherapy(P=0.011).The median progression-free survival was 15.5 months for PEM and 8.8 months for chemotherapy(P＜0.001),and the median overall survival was not reached for PEM and 26.2 months for chemotherapy(P＜0.001).Conclusion PEM treatment for advanced NSCLC demonstrates favorable survival outcomes and acceptable safety in real-world clinical practice.