Objective:To investigate the effects of long non coding RNA KCNQ1OT1(LncRNA KCNQ1OT1)on the migration and invasion of oral squamous cell carcinoma(OSCC)cells by regulating the microRNA-875-5p(miR-875-5p)/ETS like transcription factor 4(ELK4)axis.Methods:QRT-PCR was applied to detect the mRNA levels of LncRNA KCNQ1OT1,miR-875-5p,and ELK4 in OSCC cell lines(HSC-3,PE/CA-PJ15,HN13)and tissues.The dual luciferase assay was applied to detect the targeting relationship between LncRNA KCNQ1OT1 and miR-875-5p,and target relationship between miR-875-5p and ELK4.HSC-3 cells were used in control group,sh-NC group,sh-KCNQ1OT1 group,sh-KCNQ1OT1+anti-NC group,sh-KCNQ1OT1+anti-miR-875-5p group,miR-NC group,miR-875-5p mimic group,miR-875-5p mimic+pcDNA-NC group,and miR-875-5p mimic+ELK4 group.The migration and invasion abilities of HSC-3 cells were detected.Immunoblotting was applied to detect the protein expression of ELK4,MMP-2,MMP-9 and epithelial mesenchymal transition(EMT)(E-Cadherin,N-Cadherin,Vimentin).The nude mouse transplant tumor was applied to verify the effect of LncRNA KCNQ1OT1 on OSCC transplant tumors.Results:LncRNA KCNQ1OT1 and ELK4 mRNA expression increased in OSCC tissues and cancer cell lines,while miR-875-5p expression decreased(P＜0.05).Database predictions show that miR-875-5p specifically bound to LncRNAs KCNQ1OT1 and ELK4,respectively.Compared with the sh-NC group,the numbers of cell migration and cell invasion,the expression of LncRNA KCNQ1OT1,ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin in the sh-KC-NQ1OT1 group were lower,while the expression of miR-875-5p and E-Cadherin was higher(P＜0.05).Compared with the sh-KC-NQ1OT1+anti-NC group,the expression of miR-875-5p and E-Cadherin in the sh-KCNQ1OT1+anti-miR-875-5p group was lower,while the numbers of cell migration and cell invasion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin were higher(P＜0.05).Compared with the miR-NC group,the expression of miR-875-5p and E-Cadherin in the miR-875-5p mimic group was higher,while the numbers of cell migration and cell invasion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vim-entin were lower(P＜0.05).Compared with the miR-875-5p mimic+pcDNA-NC group,the numbers of cell migration and cell inva-sion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin in the miR-875-5p mimic+ELK4 group were higher,while the expression of E-Cadherin was lower(P＜0.05).The transplant tumor volume and weight of the sh-KCNQ1OT1 group were smaller than those of the sh-NC group,the mRNA and protein expression levels of LncRNA KCNQ1OT1,ELK4 were lower than those of the sh-NC group,and the expression level of miR-875-5p was higher than that of the sh-NC group(P＜0.05).Conclusion:Inhibition of LncRNA KCNQ1OT1 can target the miR-875-5p/ELK4 axis to inhibit migration,invasion,and EMT of OSCC cells.

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

Objective:To discuss the effects of microRNA(miR)-199a-5p overexpression on cell migration and apoptosis in the glioblastoma U251 cells,and to clarify the targeting regulatory relationship between miR-199a-5p and caveolin-1(CAV-1).Methods:The glioblastoma U251 cells and oligodendroglioma Hs683 cells were cultured in vitro.Western blotting method was used to detect the expression levels of CAV-1 protein in 2 kinds of cells;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-199a-5p in 2 kinds of cells.The U251 cells were divided into blank group(non-transfection),mimics NC group(transfected with empty vector),and miR-199a-5p mimics group(transfected with miR-199a-5p mimics).The Hs683 cells were divided into blank group(no transfection),inhibitor NC group(transfected with empty vector),and miR-199a-5p inhibitor group(transfected with miR-199a-5p inhibitor).RT-qPCR method was used to detect the transfection efficiency of the cells in various groups;Western blotting method was used to detect the expression levels of CAV-1 protein in the cells in various groups.TargetScan database was used to predict the binding sites between miR-199a-5p and CAV-1 in the 3'untrans lated region(3'UTR);psiCHECKTM-2-CAV-1-WT and psiCHECKTM-2-CAV-1-Mut were co-transfected with miR-199a-5p mimics and mimics NC into the U251 cells,respectively,forming psiCHECKTM-2-CAV-1-WT+mimics NC group,psiCHECKTM-2-CAV-1-WT+miR-199a-5p mimics group,psiCHECKTM-2-CAV-1-Mut+mimics NC group,and psiCHECKTM-2-CAV1-Mut+miR-199a-5p mimics group;dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-199a-5p and CAV-1;cell scratch assay was used to detect the scratch healing rates of the U251 cells in various groups;flow cytometry was used to detect the apoptotic rates of the U251 cells in various groups.Results:The Western blotting and RT-qPCR results showed that compared with Hs683 cells,the expression level of CAV-1 protein in the U251 cells was significantly decreased(P<0.05);compared with U251 cells,the expression level of miR-199a-5p in the Hs683 cells was significantly increased(P<0.01).Compared with blank group and mimics NC group,the expression level of miR-199a-5p in the U251 cells in miR-199a-5p mimics group was significantly increased(P<0.01),the expression level of CAV-1 protein in the U251 cells in miR-199a-5p mimics group was significantly decreased(P<0.05).Compared with blank group,the expression levels of miR-199a-5p in the Hs683 cells in inhibitor NC group and miR-199a-5p inhibitor group were significantly decreased(P<0.01).No significant differences were observed in the expression levels of CAV-1 protein in the Hs683 cells among various groups(P>0.05).The dual-luciferase reporter gene assay results showed that psiCHECKTM-2-CAV-1-wild type(WT)and psiCHECKTM-2-CAV-1-mutant(Mut)expression vectors were successfully constructed;compared with psiCHECKTM-2-CAV-1-WT-mimics NC group,the relative luciferase activity of WT CAV-1 in the U251 cells in psiCHECKTM-2-CAV-1-WT-miR-199a-5p mimics group was significantly decreased(P<0.01).The cell scratch assay results showed that at 12,24,and 48 h after transfection,compared with blank group,the scratch healing rate of the U251 cells in miR-199a-5p mimics group was significantly decreased(P<0.05 or P<0.01).The flow cytometry results showed that compared with blank group and mimics NC group,the apoptotic rate of the U251 cells in miR-199a-5p mimics group was significantly increased(P<0.01).Conclusion:Transfection of mature miR-199a-5p mimics into the glioblastoma U251 cells can reduce the expression of CAV-1 protein,inhibit glioma cell migration,promote apoptosis,and suppress tumorigenesis and development.The targeting relationship between miR-199a-5p and CAV-1 may represent a potential mechanism for glioma development and could serve as a potential diagnostic and therapeutic target for glioma.

Background Injury poses a serious threat to human health. As global warming continues to intensify, there is an urgent need to explore the impact of temperature changes on injury deaths. However limited research has focused on this issue. Objective To investigate the relationship between daily mean temperature change (Tm) and injury death, as well as to estimate the associated future death burden in Hunan Province. Methods We employed an individual-level, time-stratified case-crossing design to establish a conditional logistic regression model to analyze the exposure-response relationship between daily mean temperature change and injury death in Hunan Province from 2013 to 2018. Consequently, we conducted subgroup analysis of gender, age group, and injury type. Finally, we estimated the excess burden of injury death attributable to temperature changes under a sustainable development path [low emission scenario (SSP1-2.6)], regional competition path [high emission scenario (SSP3-7.0)], or fossil fuel development path [very high emission scenario (SSP5-8.5)]. Results The study collected 155577 injury deaths in Hunan Province. The results indicated that for each 1 ℃ increment in daily mean temperature, the cumulative excess risk (CER) of injury mortality among Hunan residents increased by 0.71% (95%CI: 0.53%, 0.90%). Notably, the CER for intentional injuries (1.06%, 95%CI: 0.51%, 1.61%) was higher than that for unintentional injuries (0.66%, 95%CI: 0.46%, 0.87%). Warmer temperatures were positively associated with CER of mortality due to drowning, falls, transport injuries, assaults, and suicides, while they were negatively associated with mortality due to poisoning. Compared with 2010–2019, for 2090–2099, the excess mortality rate (EMR) due to ambient temperature increase in Hunan Province under the SSP5-8.5 scenario (14.62/100000, 95%CI: 10.81/100000, 18.40/100000) would be higher than that of the SSP3-7.0 scenario (10.28/100000, 95%CI: 7.58/100000, 12.97/100000) and the SSP1-2.6 scenario (3.35/100000, 95%CI: 2.46/100000, 4.23/100000); the EMR would be around 2.5 times among men (20.64/100000, 95%CI: 14.44/100000, 26.8/100000) than that among women (8.33/100000, 95%CI: 3.98/100000, 12.6/100000). Among unintentional injuries, the leading contributors to EMR would be drowning (4.46/100000), falls (3.96/100000), and transport injuries (2.96/100000). And among intentional injuries, the EMR for suicide (2.08/100000) would be higher than that for assault (0.47/100000). Conclusion Climate warming will increase the total burden of injury-related deaths among residents in Hunan Province, especially in the absence of effective mitigation strategies and measures. It is necessary to strengthen the prevention and control of severe injuries closely related to temperature changes, such as drowning, falls, traffic, and suicides injuries, in order to reduce injury deaths associated with climate change.

As a common biliary disease, cholelithiasis has high incidence and recurrence rates, with negative impact on patients′ health and quality of life. The causes of cholelithiasis are complex, and recent studies have shown that microorganisms may be associated with the formation and recurrence of cholelithiasis, but the specific relationship between the two has not been fully elucidated. Bile microbiota may be related to the increase of incidence rate of gallstones. Different types of cholelithiasis have different microbial composition. Microorganisms may participate in the formation and recurrence of gallstones by influencing bile composition, promoting the formation of stone cores and other ways. The authors provide a review on the etiology and pathogenesis of cholelithiasis, classification of cholelithiasis, microenvironment of biliary microbiota, and the relationship between biliary microbiota and stone formation, aiming to provide new ideas and theoretical basis for the prevention and treatment of cholelithiasis.

Objective To evaluate the practical application of the Media Fill Test(MFT)in assessing aseptic compounding competency of personnel in Pharmacy Intravenous Admixture Services(PIVAS).Methods A multicenter controlled study was conducted across six tertiary hospitals(center ①-⑥)in China.Participants were divided into an inexperienced group(Group A,n=118)and an experienced group(Group B,n=118),each performing five MFT operations.Positive controls validated medium efficacy.Contamination rates and pass rates were analyzed using chi-square and Fisher's exact tests.Results Valid samples included 584 for Group A and 588 for Group B.The sample pass rate was 66.78%(390/584)in Group A and 91.67%(539/588)in Group B,while personnel pass rates were 46.15%(54/117)and 80.51%(95/118),respectively,with significant intergroup differences(both P<0.01).All centers except Center ⑥ showed significantly higher pass rates in Group B(all P<0.05).Conclusion MFT effectively differentiates technical proficiency levels and is suitable for training evaluation of novice PIVAS staff.

Objective To evaluate the practical application of the Media Fill Test(MFT)in assessing aseptic compounding competency of personnel in Pharmacy Intravenous Admixture Services(PIVAS).Methods A multicenter controlled study was conducted across six tertiary hospitals(center ①-⑥)in China.Participants were divided into an inexperienced group(Group A,n=118)and an experienced group(Group B,n=118),each performing five MFT operations.Positive controls validated medium efficacy.Contamination rates and pass rates were analyzed using chi-square and Fisher's exact tests.Results Valid samples included 584 for Group A and 588 for Group B.The sample pass rate was 66.78%(390/584)in Group A and 91.67%(539/588)in Group B,while personnel pass rates were 46.15%(54/117)and 80.51%(95/118),respectively,with significant intergroup differences(both P<0.01).All centers except Center ⑥ showed significantly higher pass rates in Group B(all P<0.05).Conclusion MFT effectively differentiates technical proficiency levels and is suitable for training evaluation of novice PIVAS staff.

Objective:To investigate the effects of long non coding RNA KCNQ1OT1(LncRNA KCNQ1OT1)on the migration and invasion of oral squamous cell carcinoma(OSCC)cells by regulating the microRNA-875-5p(miR-875-5p)/ETS like transcription factor 4(ELK4)axis.Methods:QRT-PCR was applied to detect the mRNA levels of LncRNA KCNQ1OT1,miR-875-5p,and ELK4 in OSCC cell lines(HSC-3,PE/CA-PJ15,HN13)and tissues.The dual luciferase assay was applied to detect the targeting relationship between LncRNA KCNQ1OT1 and miR-875-5p,and target relationship between miR-875-5p and ELK4.HSC-3 cells were used in control group,sh-NC group,sh-KCNQ1OT1 group,sh-KCNQ1OT1+anti-NC group,sh-KCNQ1OT1+anti-miR-875-5p group,miR-NC group,miR-875-5p mimic group,miR-875-5p mimic+pcDNA-NC group,and miR-875-5p mimic+ELK4 group.The migration and invasion abilities of HSC-3 cells were detected.Immunoblotting was applied to detect the protein expression of ELK4,MMP-2,MMP-9 and epithelial mesenchymal transition(EMT)(E-Cadherin,N-Cadherin,Vimentin).The nude mouse transplant tumor was applied to verify the effect of LncRNA KCNQ1OT1 on OSCC transplant tumors.Results:LncRNA KCNQ1OT1 and ELK4 mRNA expression increased in OSCC tissues and cancer cell lines,while miR-875-5p expression decreased(P＜0.05).Database predictions show that miR-875-5p specifically bound to LncRNAs KCNQ1OT1 and ELK4,respectively.Compared with the sh-NC group,the numbers of cell migration and cell invasion,the expression of LncRNA KCNQ1OT1,ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin in the sh-KC-NQ1OT1 group were lower,while the expression of miR-875-5p and E-Cadherin was higher(P＜0.05).Compared with the sh-KC-NQ1OT1+anti-NC group,the expression of miR-875-5p and E-Cadherin in the sh-KCNQ1OT1+anti-miR-875-5p group was lower,while the numbers of cell migration and cell invasion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin were higher(P＜0.05).Compared with the miR-NC group,the expression of miR-875-5p and E-Cadherin in the miR-875-5p mimic group was higher,while the numbers of cell migration and cell invasion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vim-entin were lower(P＜0.05).Compared with the miR-875-5p mimic+pcDNA-NC group,the numbers of cell migration and cell inva-sion,the expression of ELK4,MMP-2,MMP-9,N-Cadherin,and Vimentin in the miR-875-5p mimic+ELK4 group were higher,while the expression of E-Cadherin was lower(P＜0.05).The transplant tumor volume and weight of the sh-KCNQ1OT1 group were smaller than those of the sh-NC group,the mRNA and protein expression levels of LncRNA KCNQ1OT1,ELK4 were lower than those of the sh-NC group,and the expression level of miR-875-5p was higher than that of the sh-NC group(P＜0.05).Conclusion:Inhibition of LncRNA KCNQ1OT1 can target the miR-875-5p/ELK4 axis to inhibit migration,invasion,and EMT of OSCC cells.

As a common biliary disease, cholelithiasis has high incidence and recurrence rates, with negative impact on patients′ health and quality of life. The causes of cholelithiasis are complex, and recent studies have shown that microorganisms may be associated with the formation and recurrence of cholelithiasis, but the specific relationship between the two has not been fully elucidated. Bile microbiota may be related to the increase of incidence rate of gallstones. Different types of cholelithiasis have different microbial composition. Microorganisms may participate in the formation and recurrence of gallstones by influencing bile composition, promoting the formation of stone cores and other ways. The authors provide a review on the etiology and pathogenesis of cholelithiasis, classification of cholelithiasis, microenvironment of biliary microbiota, and the relationship between biliary microbiota and stone formation, aiming to provide new ideas and theoretical basis for the prevention and treatment of cholelithiasis.

Objective:To compare the efficacy of radial forearm flap and anterolateral thigh flap in repairing soft tissue defects after oral cancer surgery and to explore their indications.Methods:A retrospective analysis was conducted on clinical data of patients with oral cancer treated at the Department of Stomatology, Affiliated Hospital of Nantong University, from May 2019 to February 2023. Patients were divided into two groups based on the repair method: the radial forearm flap group and the anterolateral thigh flap group. The groups were compared in the following aspects. (1) Surgical parameters including defect area after oral cancer resection, flap area, flap preparation time, operation time, and length of hospital stay. (2) Inflammatory markers (interleukin-6 and C-reactive protein levels) measured 1 day before surgery and 1 day after surgery. (3) Flap survival rate was calculated. (4) Complication rates was calculated in the flap donor area and infection rates in the oral recipient area within 6 months postoperatively. (5) Six months postoperatively, the patient’s oral function was assessed by a physician using the University of Washington quality of life scale (UW-QOL). The evaluation included assessments of oral opening, speech, and eating functions. Each parameter was scored on a scale of 0 to 10 (higher scores indicated better recovery). (6) Quality of life was assessed using the 36-item short form health survey scale(SF-36) at 2, 4 and 6 months postoperatively, with scores ranging from 0 to 100 (higher scores indicated better quality of life). (7) Patient satisfaction was assessed at 6 months postoperatively, with satisfaction levels categorized as satisfied, basically satisfied, and dissatisfied. The satisfaction rate was calculated as (satisfied + basically satisfied ) cases / total cases in each group × 100%. Statistical analysis was performed using SPSS 22.0. Measurement data were expressed as Mean±SD, and comparisons between groups were conducted using t-tests. Count data were expressed as cases and (or) percentages, and comparisons were made using chi-square test. P<0.05 was considered statistically significant. Results:The radial forearm flap group included 48 cases (32 males, 16 females), aged (49.3±5.0) years, with a body mass index (BMI) of (23.0±1.1) kg/m 2 and a disease course of (6.5±2.1) months. The group had 21 cases of tongue cancer, 12 of floor of mouth cancer, and 15 of buccal cancer, including 40 squamous cell carcinomas and 8 basal cell carcinomas. The anterolateral thigh flap group included 32 cases (20 males, 12 females), aged (50.1±5.0) years, with a BMI of (23.0±1.0) kg/m 2 and a disease course of (7.0±2.2) months. The group had 16 cases of tongue cancer, 7 cases of floor of mouth cancer, and 9 cases of buccal cancer, including 27 squamous cell carcinomas and 5 basal cell carcinomas. There were no significant differences in gender, age, BMI, disease course, tumor location, or pathological type between the two groups ( P>0.05). The defect area after oral cancer resection was smaller in the radial forearm flap group[ (39.0±1.3) cm 2 ] compared to the anterolateral thigh flap group[ (40.3±2.2) cm 2] ( t=3.32, P=0.001). There were no significant differences in flap area, flap preparation time, or length of hospital stay between the two groups ( P>0.05). The operation time was shorter in the radial forearm flap group [(5.1±1.1) h] compared to the anterolateral thigh flap group [(6.8±2.8) h] ( t=0.26, P<0.001). There were no significant differences in interleukin-6 and C-reactive protein levels between the two groups 1 day before surgery and 1 day after surgery ( P>0.05). The flap survival rates were 97.9% (47/48) in the radial forearm flap group and 93.8% (30/32) in the anterolateral thigh flap group, with no significant difference( P>0.05). Postoperative donor site complications mainly included infection, pigmentation, itching, etc. The overall incidence of complications in the donor site of the radial forearm flap [33.3% (16/48)] was higher than that in the anterolateral thigh flap group [12.5% (4/32)], and the difference was statistically significant ( χ2=4.44, P=0.035). There was no significant difference in infection rates in the oral recipient area between the two groups ( P>0.05). Six months postoperatively, the average scores for oral opening, speech, and eating functions were above 7 in both groups, with no significant differences ( P>0.05). Quality of life scores improved over time in both groups, with average scores above 90 at 6 months postoperatively, and no significant differences at any time point ( P>0.05). The patient satisfaction rate was 91.7% (44/48) in the radial forearm flap group and 90.6% (29/32) in the anterolateral thigh flap group, with no significant difference ( P>0.05). Conclusion:Both radial forearm flap and anterolateral thigh flap can effectively repair soft tissue defects after oral cancer resection, significantly improving patients’oral function. The anterolateral thigh flap provides sufficient tissue volume and is suitable for patients with larger defect areas. The radial forearm flap is suitable for patients with a smaller defect area after oral cancer resection. Its surgical procedure is relatively less complex and offers an advantage in reducing surgery time. However, the donor site complications are higher with the radial forearm flap compared to the anterolateral thigh flap.