Objective To explore the application effect of ultrasound-assisted localization in spinal anesthesia for elderly patients with hip fractures.Methods A total of 114 elderly patients undergoing hip fracture surgery with spinal anesthesia in our hospital from September 2022 to June 2024 were enrolled.The random number table method was applied to allocate participants into two groups:surface anatomical landmark localization group and ultrasound-assisted localization group,with 57 patients in each group.Both groups of patients underwent spinal anesthesia via the paramedian approach for puncture.The first-attempt puncture success rate,success rate of the initial puncture site,number of skin punctures,number of adjusted epidural needle direction,positioning time of the puncture point,puncture time(puncture success time,anesthesia time,and total time),adverse events during the puncture process(nerve hypersensitivity and accidental puncture of the dura mater),spinal anesthesia related complications(back pain,headache,and nerve injury),patient satisfaction and hospital stay were observed and compared between the two groups.The quality of early postoperative recovery was evaluated by using the 15-Item Quality of Recovery(QOR-15)scale at 24 h after surgery.Results The first-attempt puncture success rate in the ultrasound-assisted localization group was 71.9％(41/57),which was significantly higher than that in the surface anatomical landmark localization group[28.1％(16/57),x2=21.930,P=0.000].The success rate of the initial puncture site in the ultrasound-assisted localization group was 87.7％(50/57),which was significantly higher than that in the surface anatomical landmark localization group[54.4％(31/57),x2=15.396,P=0.000].The number of skin punctures and adjusted epidural needle direction in the ultrasound-assisted localization group were 1(1,1)and 0(0,1)times,while in the surface anatomical landmark localization group were 2(1,3)and 3(0,5)times,with statistically significant differences(all P=0.000).The puncture positioning time in the ultrasound-assisted localization group was 2.2(1.7,3.2)min,which was significantly longer than that in the surface anatomical landmark localization group[0.8(0.5,1.2)min,Z=-8.418,P=0.000].The puncture success time,anesthesia time,and total time of the ultrasound-assisted localization group were 1.9(1.7,2.3),4.9(4.3,5.3),and 7.1(6.3,8.7)min,while of the surface anatomical landmark localization group were 5.1(2.3,8.0),7.9(5.7,11.0),and 8.6(6.6,12.0)min,with statistically significant differences(all P＜0.05).There were no statistically significant differences in the incidence of nerve hypersensitivity during operation(3.5％vs.7.0％,x2=0.176,P=0.675),accidental puncture of the dura mater by epidural needles(0％vs.3.5％,P=0.496),postoperative headache(0％vs.3.5％,P=0.496),and back pain(1.8％vs.10.5％,x2=2.435,P=0.119)between the two groups.Both groups of patients showed no symptoms of nerve injury after surgery.There were no statistically significant differences in QOR-15 scores at 24 h postoperatively[119(115,124)points vs.116(112,121)points,Z=-1.858,P=0.063]and length of hospital stay[10.0(9.0,12.5)dvs.10.0(8.0,13.0)d,Z=-0.043,P=0.966]between the two groups.The satisfaction of patients in the ultrasound-assisted localization group was significantly higher than that in the surface anatomical landmark localization group(P=0.004).Conclusion Application of ultrasound-guided localization in spinal anesthesia for elderly patients with hip fractures significantly improves the first-attempt puncture and initial puncture site success rates,reduces skin puncture attempts and adjustments of the epidural needle direction,shortens time of anesthetic procedure,and enhances patient satisfaction,making it highly recommendable for clinical use.

Background and purpose:Gallic acid(GA)induces tumor cells apoptosis and inhibits angiogenesis.Beyond directly attacking tumor cells,another crucial aspect of GA is its ability to modulate and enhance immune system function.For example,it can improve T cell metabolism,alleviate T cell exhaustion,and promote the formation of memory T cell phenotypes.Although several chimeric antigen receptor T(CAR-T)cells products have gained market approval,the technology still faces significant challenges.These limitations include off-target effects,a predisposition to T cell exhaustion and so on.Moreover,similar to exhaustion,cellular senescence is a major hindrance that impairs T cell function.This study aimed to investigate the effects of GA on the anti-tumor function of CAR-T cells both in vitro and in vivo.We further evaluated the impact of GA on CAR-T cells senescence and memory phenotypes,as well as the impact of GA and CAR-T cells on immune cell infiltration within the tumor microenvironment(TME).Methods:Second-generation CAR targeting mouse glypican 3(GPC3)and human epidermal growth factor receptor 2(HER2)were constructed to generate CAR-T cells.CAR-T cells were co-cultured with GA at a concentration of 5 μg/mL,and flow cytometry was used to assess the senescence status and memory phenotype of CAR-T cells and their killing ability against tumor cells at different effector-to-target ratios.Senescence markers included p53,p21,γ-H2AX and senescence-associated β-galactosidase(SA-β-gal),while CCR7 served as the memory phenotype marker.A subcutaneous tumor model was established to explore the effects of GA on the anti-tumor function of CAR-T cells and immune cell infiltration within the TME.Results:We successfully generated human HER2 and murine GPC3 CAR-T cells,achieving a purity of 30%-50%.GA enhanced the in vitro killing ability of CAR-T cells targeting mouse GPC3 and human HER2(P＜0.001)at different E:T ratios,delayed the senescence of mouse GPC3 CAR-T cells(p53,p21,γ-H2AX,P＜0.05;SA-β-gal,P＜0.001;CCR7,P＜0.001).And GA promoted the differentiation of CAR-T cells toward a memory phenotype(P＜0.001).Additionally,GPC3 CAR-T cells inhibited tumor cell growth(P＜0.05),prolonged mouse survival(P＜0.001),and enhanced the infiltration capacity of CAR-cells(P＜0.001)and endogenous immune cells[CD4+T cells,P＜0.05;CD8+T cells,P＜0.01;natural killer(NK)cells,P＜0.01].Conclusion:GA can enhance the cytotoxic activity of CAR-T cells in vitro,and delay the senescence of CAR-T cells.Furthermore,by modulating TME,GA improved immune cell infiltration,thereby augmenting the overall anti-tumor efficacy of CAR-T cells.

BACKGROUND: Bile acids (BAs) facilitate the progression of gastric intestinal metaplasia (GIM). Long non-coding RNAs (lncRNAs) dysregulation was observed along with the initiation of gastric cancer. However, how lncRNAs function in GIM remains unclear. This study aimed to explore the role and mechanism of lncRNA PVT1 in GIM, and provide a potential therapeutic target for GIM treatment. METHODS: We employed RNA sequencing (RNA-seq) to screen dysregulated lncRNAs in gastric epithelial cells after BA treatment. Bioinformatics analysis was conducted to reveal the regulatory mechanism. PVT1 expression was detected in 21 paired biopsies obtained under endoscopy. Overexpressed and knockdown cell models were established to explore gene functions in GIM. Molecular interactions were validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (Ch-IP). The levels of relative molecular expression were detected in GIM tissues. RESULTS: We confirmed that lncRNA PVT1 was upregulated in BA-induced GIM model. PVT1 promoted the expression of intestinal markers such as CDX2 , KLF4 , and HNF4α . Bioinformatics analysis revealed that miR-34b-5p was a putative target of PVT1 . miR-34b-5p mimics increased CDX2 , KLF4 , and HNF4α levels. Restoration of miR-34b-5p decreased the pro-metaplastic effect of PVT1 . The interactions between PVT1 , miR-34b-5p, and the downstream target HNF4α were validated. Moreover, HNF4α could transcriptionally activated PVT1 , sustaining the GIM phenotype. Finally, the activation of the PVT1 /miR-34b-5p/ HNF4α loop was detected in GIM tissues. CONCLUSIONS BAs facilitate GIM partially via a PVT1/miR-34b-5p/HNF4α positive feedback loop. PVT1 may become a novel target for blocking the continuous development of GIM and preventing the initiation of gastric cancer in patients with bile reflux.
Humans ; RNA, Long Noncoding/metabolism* ; MicroRNAs/metabolism* ; Hepatocyte Nuclear Factor 4/genetics* ; Bile Acids and Salts ; Kruppel-Like Factor 4 ; Metaplasia/metabolism*

Objective To explore the application effect of ultrasound-assisted localization in spinal anesthesia for elderly patients with hip fractures.Methods A total of 114 elderly patients undergoing hip fracture surgery with spinal anesthesia in our hospital from September 2022 to June 2024 were enrolled.The random number table method was applied to allocate participants into two groups:surface anatomical landmark localization group and ultrasound-assisted localization group,with 57 patients in each group.Both groups of patients underwent spinal anesthesia via the paramedian approach for puncture.The first-attempt puncture success rate,success rate of the initial puncture site,number of skin punctures,number of adjusted epidural needle direction,positioning time of the puncture point,puncture time(puncture success time,anesthesia time,and total time),adverse events during the puncture process(nerve hypersensitivity and accidental puncture of the dura mater),spinal anesthesia related complications(back pain,headache,and nerve injury),patient satisfaction and hospital stay were observed and compared between the two groups.The quality of early postoperative recovery was evaluated by using the 15-Item Quality of Recovery(QOR-15)scale at 24 h after surgery.Results The first-attempt puncture success rate in the ultrasound-assisted localization group was 71.9％(41/57),which was significantly higher than that in the surface anatomical landmark localization group[28.1％(16/57),x2=21.930,P=0.000].The success rate of the initial puncture site in the ultrasound-assisted localization group was 87.7％(50/57),which was significantly higher than that in the surface anatomical landmark localization group[54.4％(31/57),x2=15.396,P=0.000].The number of skin punctures and adjusted epidural needle direction in the ultrasound-assisted localization group were 1(1,1)and 0(0,1)times,while in the surface anatomical landmark localization group were 2(1,3)and 3(0,5)times,with statistically significant differences(all P=0.000).The puncture positioning time in the ultrasound-assisted localization group was 2.2(1.7,3.2)min,which was significantly longer than that in the surface anatomical landmark localization group[0.8(0.5,1.2)min,Z=-8.418,P=0.000].The puncture success time,anesthesia time,and total time of the ultrasound-assisted localization group were 1.9(1.7,2.3),4.9(4.3,5.3),and 7.1(6.3,8.7)min,while of the surface anatomical landmark localization group were 5.1(2.3,8.0),7.9(5.7,11.0),and 8.6(6.6,12.0)min,with statistically significant differences(all P＜0.05).There were no statistically significant differences in the incidence of nerve hypersensitivity during operation(3.5％vs.7.0％,x2=0.176,P=0.675),accidental puncture of the dura mater by epidural needles(0％vs.3.5％,P=0.496),postoperative headache(0％vs.3.5％,P=0.496),and back pain(1.8％vs.10.5％,x2=2.435,P=0.119)between the two groups.Both groups of patients showed no symptoms of nerve injury after surgery.There were no statistically significant differences in QOR-15 scores at 24 h postoperatively[119(115,124)points vs.116(112,121)points,Z=-1.858,P=0.063]and length of hospital stay[10.0(9.0,12.5)dvs.10.0(8.0,13.0)d,Z=-0.043,P=0.966]between the two groups.The satisfaction of patients in the ultrasound-assisted localization group was significantly higher than that in the surface anatomical landmark localization group(P=0.004).Conclusion Application of ultrasound-guided localization in spinal anesthesia for elderly patients with hip fractures significantly improves the first-attempt puncture and initial puncture site success rates,reduces skin puncture attempts and adjustments of the epidural needle direction,shortens time of anesthetic procedure,and enhances patient satisfaction,making it highly recommendable for clinical use.

Background and purpose:Gallic acid(GA)induces tumor cells apoptosis and inhibits angiogenesis.Beyond directly attacking tumor cells,another crucial aspect of GA is its ability to modulate and enhance immune system function.For example,it can improve T cell metabolism,alleviate T cell exhaustion,and promote the formation of memory T cell phenotypes.Although several chimeric antigen receptor T(CAR-T)cells products have gained market approval,the technology still faces significant challenges.These limitations include off-target effects,a predisposition to T cell exhaustion and so on.Moreover,similar to exhaustion,cellular senescence is a major hindrance that impairs T cell function.This study aimed to investigate the effects of GA on the anti-tumor function of CAR-T cells both in vitro and in vivo.We further evaluated the impact of GA on CAR-T cells senescence and memory phenotypes,as well as the impact of GA and CAR-T cells on immune cell infiltration within the tumor microenvironment(TME).Methods:Second-generation CAR targeting mouse glypican 3(GPC3)and human epidermal growth factor receptor 2(HER2)were constructed to generate CAR-T cells.CAR-T cells were co-cultured with GA at a concentration of 5 μg/mL,and flow cytometry was used to assess the senescence status and memory phenotype of CAR-T cells and their killing ability against tumor cells at different effector-to-target ratios.Senescence markers included p53,p21,γ-H2AX and senescence-associated β-galactosidase(SA-β-gal),while CCR7 served as the memory phenotype marker.A subcutaneous tumor model was established to explore the effects of GA on the anti-tumor function of CAR-T cells and immune cell infiltration within the TME.Results:We successfully generated human HER2 and murine GPC3 CAR-T cells,achieving a purity of 30%-50%.GA enhanced the in vitro killing ability of CAR-T cells targeting mouse GPC3 and human HER2(P＜0.001)at different E:T ratios,delayed the senescence of mouse GPC3 CAR-T cells(p53,p21,γ-H2AX,P＜0.05;SA-β-gal,P＜0.001;CCR7,P＜0.001).And GA promoted the differentiation of CAR-T cells toward a memory phenotype(P＜0.001).Additionally,GPC3 CAR-T cells inhibited tumor cell growth(P＜0.05),prolonged mouse survival(P＜0.001),and enhanced the infiltration capacity of CAR-cells(P＜0.001)and endogenous immune cells[CD4+T cells,P＜0.05;CD8+T cells,P＜0.01;natural killer(NK)cells,P＜0.01].Conclusion:GA can enhance the cytotoxic activity of CAR-T cells in vitro,and delay the senescence of CAR-T cells.Furthermore,by modulating TME,GA improved immune cell infiltration,thereby augmenting the overall anti-tumor efficacy of CAR-T cells.

Objective:This study aims to investigate the current application and the level of knowledge of intraoperative facial nerve monitoring among medical staff in China.Methods:A comprehensive online questionnaire was conducted among medical professionals across different regions in China from October 2022 to February 2023. The survey exclusively targeted departments specializing in otolaryngology, head and neck surgery, neurosurgery, and oral and maxillofacial surgery. The questionnaire covered various aspects including general information, intraoperative facial nerve monitoring practices, training history, indications for monitoring, parameters used during monitoring procedures, as well as factors influencing its implementation.Results:A total of 417 participants from 31 provincial, municipal, and autonomous regions were included. Intraoperative facial nerve monitoring was found to be implemented in 227 (54.4%,227/417) repondents of 53 institutions (24.9%, 53/213). The top three indications for implementing this technique were acoustic neuroma, parotid gland surgery, and modified middle ear surgery (mastoidectomy). Herein 81.1%(184/227) medical staff involved in intraoperative facial nerve monitoring had received relevant training, 57.3%(130/227)-92.1%(209/227) reported a lack of clear description regarding recording thresholds, stimulation currents/frequencies/wave widths.Conclusion:The majority of the institutions surveyed have not yet adopted intraoperative facial nerve monitoring. Furthermore, significant gaps concerning the procedure exist. It is imperative to establish standards or guidelines to promote its better development and application.

Diabetes, hypertension, and dyslipidemia, collectively referred to the " Three Highs, " represent increasingly prevalent metabolic risk factors in China. Many individuals experience all three conditions concurrently, significantly heightening the risk of cardiovascular disease and mortality. Although the National Metabolic Management Center(MMC) has been established for over eight years and has its unique features, the awareness, treatment, and control rates of these diseases in China remain low, and the efficiency of community management is insufficient. According to the previous two editions of management guidelines and the most recent domestic and international diagnostic and treatment guidelines, this paper conducts an in-depth analysis of the operational experience and management strategies of the MMC. Its aim is to improve the efficiency of grassroots MMC mode management for " Three Highs" patients and ensure that patients receive more standardized management.

Objective:To construct dual-targeting(CD38 and CD138)chimeric antigen receptor(CAR)gene-modified T cells(CD38/CD138 CAR-T cells)and explore their in vitro cytotoxicity against multiple myeloma(MM)cells.Methods:Based on the high expression of CD38 and CD138 antigens in MM cells,CD38 CAR-T cells and CD138 CAR-T cells targeting CD38 and CD138 respectively,and CD38/CD138 dual-targeted CAR-T cells targeting both CD38 and CD138 were constructed using CAR-T cell technology.The experimental groups included untreated T cells,CD38 CAR-T,CD138 CAR-T,and CD38/CD138 CAR-T cells.The phenotype of CAR-T cells was detected by flow cytometry.The cytotoxicity of various CAR-T cells against MM cells(RPMI8226 and U266)was assessed using the LDH release assay.Results:Three types of CAR-T cells,CD38 CAR-T,CD138 CAR-T,and CD38/CD138 CAR-T cells,were successfully constructed.The CD38/CD138 CAR-T cells tended to differentiate into a memory phenotype,expressing higher levels of proliferation marker(CD25),activation marker(CD27),and lower levels of exhaustion markers(PD-1,CTLA-4,TIM-3)(all P<0.001).Moreover,CD38/CD138 CAR-T cells were less prone to exhaustion and senescence,and expressed lower levels of r-H2AX,p-p53,p21,and p16 proteins(all P<0.01).Under different effector-to-target cell ratios,CD38/CD138 CAR-T cells exhibited stronger cytotoxic effects against RPMI8226 and U266 cells compared to CD38 CAR-T and CD138 CAR-T cells(all P<0.001).Conclusion:CD38/CD138 CAR-T cells targeting both CD38 and CD138 demonstrate an optimal phenotype and enhanced anti-tumor activity in vitro,offering promising potential for immunotherapy in multiple myeloma.

Diabetes, hypertension, and dyslipidemia, collectively referred to the " Three Highs, " represent increasingly prevalent metabolic risk factors in China. Many individuals experience all three conditions concurrently, significantly heightening the risk of cardiovascular disease and mortality. Although the National Metabolic Management Center(MMC) has been established for over eight years and has its unique features, the awareness, treatment, and control rates of these diseases in China remain low, and the efficiency of community management is insufficient. According to the previous two editions of management guidelines and the most recent domestic and international diagnostic and treatment guidelines, this paper conducts an in-depth analysis of the operational experience and management strategies of the MMC. Its aim is to improve the efficiency of grassroots MMC mode management for " Three Highs" patients and ensure that patients receive more standardized management.

Humans ; Bile Reflux/complications* ; Cross-Sectional Studies ; Risk Factors ; Gastric Mucosa