Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO₂) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.

T7 RNA polymerase (T7 RNAP) is one of the simplest known RNA polymerases. Its unique structural features make it a critical model for studying the mechanisms of RNA synthesis. This review systematically examines the static crystal structure of T7 RNAP, beginning with an in-depth examination of its characteristic “thumb”, “palm”, and “finger” domains, which form the classic “right-hand-like” architecture. By detailing these structural elements, this review establishes a foundation for understanding the overall organization of T7 RNAP. This review systematically maps the functional roles of secondary structural elements and their subdomains in transcriptional catalysis, progressively elucidating the fundamental relationships between structure and function. Further, the intrinsic flexibility of T7 RNAP and its applications in research are also discussed. Additionally, the review presents the structural diagrams of the enzyme at different stages of the transcription process, and through these diagrams, it provides a detailed description of the complete transcription process of T7 RNAP. By integrating structural dynamics and kinetics analyses, the review constructs a comprehensive framework that bridges static structure to dynamic processes. Despite its advantages, T7 RNAP has a notable limitation: it generates double-stranded RNA (dsRNA) as a byproduct. The presence of dsRNA not only compromises the purity of mRNA products but also elicits nonspecific immune responses, which pose significant challenges for biotechnological and therapeutic applications. The review provides a detailed exploration of the mechanisms underlying dsRNA formation during T7 RNAP catalysis, reviews current strategies to mitigate this issue, and highlights recent progress in the field. A key focus is the semi-rational design of T7 RNAP mutants engineered to minimize dsRNA generation and enhance catalytic performance. Beyond its role in transcription, T7 RNAP exhibits rapid development and extensive application in fields, including gene editing, biosensing, and mRNA vaccines. This review systematically examines the structure-function relationships of T7 RNAP, elucidates the mechanisms of dsRNA formation, and discusses engineering strategies to optimize its performance. It further explores the engineering optimization and functional expansion of T7 RNAP. Furthermore, this review also addresses the pressing issues that currently need resolution, discusses the major challenges in the practical application of T7 RNAP, and provides an outlook on potential future research directions. In summary, this review provides a comprehensive analysis of T7 RNAP, ranging from its structural architecture to cutting-edge applications. We systematically examine: (1) the characteristic right-hand domains (thumb, palm, fingers) that define its minimalistic structure; (2) the structure-function relationships underlying transcriptional catalysis; and (3) the dynamic transitions during the complete transcription cycle. While highlighting T7 RNAP’s versatility in gene editing, biosensing, and mRNA vaccine production, we critically address its major limitation—dsRNA byproduct formation—and evaluate engineering solutions including semi-rationally designed mutants. By synthesizing current knowledge and identifying key challenges, this work aims to provide novel insights for the development and application of T7 RNAP and to foster further thought and progress in related fields.

Objective： To analyze the symptoms, diagnosis and treatment of upper urinary tract calculi patients combined with mild and moderate benign prostatic hyperplasia (BPH) after ureteral stent implantation. Methods： One hundred and six BPH patients who were hospitalized for upper urinary tract calculi and had ureteral stents retained from January 2019 to December 2022 were selected and divided into 2 weeks group and 4 weeks group according to the time of removal of ureteral stents after surgery. Their general clinical data were analyzed and compared. International Prostatic Symptom Scale (IPSS), postoperative ureteral Stent Symptom Questionnaire (USSQ), and incidence of adverse events after ureteral stent removal were recorded before and after removal. Results： The scores of IPSS were significantly increased in all patients, and symptoms in urinary tract had improved significantly after discharge (P<0.05). Compared with the 2 weeks group, the USSQ score of the 4 weeks group was significantly increased (P<0.05). And no significant adverse event was observed in the 2 weeks group after the removal of ureteral sten. Conclusion： IPSS score and USSQ score increased significantly during stent implantation in BPH patients with lithiasis. And complications increased significantly over time. Following thorough clinical assessment, early ureteral stent removal demonstrates both safety and efficacy, representing an optimal therapeutic approach in selected cases.
Humans ; Male ; Prostatic Hyperplasia/surgery* ; Stents ; Ureter/surgery* ; Aged ; Middle Aged ; Urinary Calculi/surgery* ; Ureteral Calculi/surgery*

Schizophrenia (SZ) stands as a severe psychiatric disorder. This study applied diffusion tensor imaging (DTI) data in conjunction with graph neural networks to distinguish SZ patients from normal controls (NCs) and showcases the superior performance of a graph neural network integrating combined fractional anisotropy and fiber number brain network features, achieving an accuracy of 73.79% in distinguishing SZ patients from NCs. Beyond mere discrimination, our study delved deeper into the advantages of utilizing white matter brain network features for identifying SZ patients through interpretable model analysis and gene expression analysis. These analyses uncovered intricate interrelationships between brain imaging markers and genetic biomarkers, providing novel insights into the neuropathological basis of SZ. In summary, our findings underscore the potential of graph neural networks applied to multimodal DTI data for enhancing SZ detection through an integrated analysis of neuroimaging and genetic features.
Humans ; Schizophrenia/pathology* ; Diffusion Tensor Imaging/methods* ; Male ; Female ; Adult ; Brain/metabolism* ; Young Adult ; Middle Aged ; White Matter/pathology* ; Gene Expression ; Nerve Net/diagnostic imaging* ; Graph Neural Networks

Objective:To observe the effect of human mesenchymal stem cells(hMSCs)conditioned medium(CM)co-cultured with the human liposarcoma SW872 cells on the proliferation and migration of the tumor cells,and to discuss the effect of hMSCs CM on the liposarcoma cells and the possible mechanism.Methods:The hMSCs were cultured in vitro and transfected with either lentiviral vector control shNS(control group)or lentiviral shRNA targeting Yes-associated protein(YAP)(shYAP-hMSCs group)by lentiviral methods.The expression levels of YAP mRNA and protein in the hMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The CM was then harvested.The SW872 cells were cultured in vitro and divided into control group(normal culture),hMSCs CM group,and shYAP-hMSCs CM group.The proliferation activities of the cells in various groups were detected by CCK-8 assay;the apoptotic rates of the cells in various groups were detected by flow cytometry;the scratch healing rates of the cells in various groups were detected by cell scratch assay;the expression levels of YAP,matrix metallopeptidase-9(MMP-9),and cyclin D1 proteins in the cells in various groups were detected by Western blotting method.Results:Compared with control group,the expression levels of YAP mRNA and protein in the cells in shYAP-hMSCs group were decreased(P＜0.01),indicating the successful establishment of a stable transfected cell line.The CCK-8 assay results showed that compared with control group,the proliferation activity of the cells in hMSCs CM group was increased(P＜0.05),and the proliferation activity of the cells in shYAP-hMSCs CM group was decreased(P＜0.01).The flow cytometry results showed that compared with control group,there was no significant change in the apoptotic rate of the cells in hMSCs CM group(P＞0.05),while the apoptotic rate of the cells in shYAP-hMSCs CM group was increased(P＜0.01).The cell scratch assay results showed that compared with control group,the scratch healing rate of the cells in hMSCs CM group was increased(P＜0.05),and the scratch healing rate of the cells in shYAP-hMSCs CM group was decreased(P＜0.01).The Western blotting results showed that compared with control group,there were no significant differences in the expression levels of YAP,MMP-9,and cyclin D1 proteins in the cells in hMSCs CM group(P＞0.05),while the expression levels of YAP,MMP-9,and cyclin D1 proteins in the cells in shYAP-hMSCs group were decreased(P＜0.05 or P＜0.01).Conclusion:The hMSCs regulate the proliferation and migration of the human liposarcoma SW872 cells,and its mechanism may be related to the expression of YAP.

Objective:To establish a cell line stably expressing the transient receptor potential melastatin 2(TRPM2)channel for screening TRPM2 inhibitors based on PiggyBac transposition system.Methods:A plasmid PiggyBac-human TRPM2(pPB-hTRPM2)eukaryotic expression vector was constructed using PiggyBac transposition system.The plasmid and a helper plasmid were co-transfected into HEK293T cells to express TRPM2,which was identified by fluorescence and patch-clamp assays.The high throughput screening performance was assessed with the Z'factor.Calcium imaging and patch clamp techniques were employed to assess the initial activity of eleven compound molecules,confirming the inhibitory effects of the primary molecules on TRPM2.The protective effect of the screened compounds on damaged cells was validated using the oxygen-glucose deprivation/reperfusion(OGD/R)injury model and CCK-8 kit.The level of cellular reactive oxygen species(ROS)was detected by flow cytometry.The neuroprotective effects of the compounds were evaluated using a transient middle cerebral artery occlusion(tMCAO)mouse model.Results:The HEK293T cells transfected with pPB-hTRPM2-EGFP showed high TRPM2 expression.Puromycin-resistant cells,selected through screening,exhibited robust fluorescence.Whole-cell patch results revealed that induced cells displayed classical TRPM2 current characteristics comparable to the control group,showing no significant differences(P＞0.05).With a Z'factor of 0.5416 in calcium imaging,the model demonstrated suitability for high-throughput screening of TRPM2 inhibitors.Calcium imaging and electrophysiological experiments indicated that compound 6 significantly inhibited the TRPM2 channel.Further experiments showed that 1.0 μmol/L of compound 6 enhanced cell viability(P＜0.05)and reduced the level of ROS(P＜0.05)of SH-SY5Y under OGD/R injury.0.3 and 1.0 mg/kg of compound 6 reduced the cerebral infarction volume in tMCAO mice(both P＜0.05).Conclusions:A stable TRPM2 gene expressing cell line has been successfully established using PiggyBac gene editing in this study.TRPM2 channel inhibitors were screened through calcium imaging and patch clamp techniques,and an inhibitor compound 6 was identified.This compound can alleviate cell damage after OGD/R by reducing cellular ROS levels and has a protective effect against cerebral ischemia-reperfusion injury in mice.

Objective:The clinical data of prenatal stillbirth were analyzed in order to increase the understand-ing of the causes of stillbirth.Methods:Prenatal stillbirth cases that terminated pregnancy in Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University from January 2018 to December 2022 were col-lected,and the distribution characteristics of clinical data and the stillbirth causes were analyzed.The causes of death were classified according to the standards developed by the Stillbirth Collaborative Research Network(SCRN)in the United States,and they were divided into clear cause-of-death group and unknown cause-of-death group.The different characteristics of the two groups were compared and analyzed.Results:There were 210 ca-ses of prenatal stillbirth during the study period,and 104 cases met the inclusion criteria.Among them,33 cases(31.7%)had autopsy results,39 cases(37.5%)had genetic results,and 75 cases(72.1%)had placental pathol-ogy.According to the classification of SCRN standard,55 cases(52.9%)were probably related to the cause of death,33 cases(31.7%)were classified as possible,13 cases(12.5%)were probably unrelated,and 3 cases(2.9%)could not be attributed to the cause of death,that is,84.6%(88 cases)in the clear cause of death group and 15.4%(16 cases)in the unknown cause of death group.The rate of placental pathological examination in the clear cause of death group was significantly higher than that in the unknown cause of death group(78.4%).In the classification of causes of death,placental pathological changes accounted for the largest proportion,account-ing for 26.9%(28 cases),followed by pregnancy complications accounting for 25.0%(26 cases),and 15.4%of the cases were still unexplained.Conclusions:Placental pathology is of great significance for clarifying the cause of stillbirth.It is feasible to use SCRN to classify the etiology of stillbirth.Pathological placental conditions account for a relatively high proportion in the classification of stillbirth causes.It is recommended that each case of stillbirth placenta should undergo pathological examination.

Objective:The clinical data of prenatal stillbirth were analyzed in order to increase the understand-ing of the causes of stillbirth.Methods:Prenatal stillbirth cases that terminated pregnancy in Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University from January 2018 to December 2022 were col-lected,and the distribution characteristics of clinical data and the stillbirth causes were analyzed.The causes of death were classified according to the standards developed by the Stillbirth Collaborative Research Network(SCRN)in the United States,and they were divided into clear cause-of-death group and unknown cause-of-death group.The different characteristics of the two groups were compared and analyzed.Results:There were 210 ca-ses of prenatal stillbirth during the study period,and 104 cases met the inclusion criteria.Among them,33 cases(31.7%)had autopsy results,39 cases(37.5%)had genetic results,and 75 cases(72.1%)had placental pathol-ogy.According to the classification of SCRN standard,55 cases(52.9%)were probably related to the cause of death,33 cases(31.7%)were classified as possible,13 cases(12.5%)were probably unrelated,and 3 cases(2.9%)could not be attributed to the cause of death,that is,84.6%(88 cases)in the clear cause of death group and 15.4%(16 cases)in the unknown cause of death group.The rate of placental pathological examination in the clear cause of death group was significantly higher than that in the unknown cause of death group(78.4%).In the classification of causes of death,placental pathological changes accounted for the largest proportion,account-ing for 26.9%(28 cases),followed by pregnancy complications accounting for 25.0%(26 cases),and 15.4%of the cases were still unexplained.Conclusions:Placental pathology is of great significance for clarifying the cause of stillbirth.It is feasible to use SCRN to classify the etiology of stillbirth.Pathological placental conditions account for a relatively high proportion in the classification of stillbirth causes.It is recommended that each case of stillbirth placenta should undergo pathological examination.

Objective:The clinical data of prenatal stillbirth were analyzed in order to increase the understand-ing of the causes of stillbirth.Methods:Prenatal stillbirth cases that terminated pregnancy in Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University from January 2018 to December 2022 were col-lected,and the distribution characteristics of clinical data and the stillbirth causes were analyzed.The causes of death were classified according to the standards developed by the Stillbirth Collaborative Research Network(SCRN)in the United States,and they were divided into clear cause-of-death group and unknown cause-of-death group.The different characteristics of the two groups were compared and analyzed.Results:There were 210 ca-ses of prenatal stillbirth during the study period,and 104 cases met the inclusion criteria.Among them,33 cases(31.7%)had autopsy results,39 cases(37.5%)had genetic results,and 75 cases(72.1%)had placental pathol-ogy.According to the classification of SCRN standard,55 cases(52.9%)were probably related to the cause of death,33 cases(31.7%)were classified as possible,13 cases(12.5%)were probably unrelated,and 3 cases(2.9%)could not be attributed to the cause of death,that is,84.6%(88 cases)in the clear cause of death group and 15.4%(16 cases)in the unknown cause of death group.The rate of placental pathological examination in the clear cause of death group was significantly higher than that in the unknown cause of death group(78.4%).In the classification of causes of death,placental pathological changes accounted for the largest proportion,account-ing for 26.9%(28 cases),followed by pregnancy complications accounting for 25.0%(26 cases),and 15.4%of the cases were still unexplained.Conclusions:Placental pathology is of great significance for clarifying the cause of stillbirth.It is feasible to use SCRN to classify the etiology of stillbirth.Pathological placental conditions account for a relatively high proportion in the classification of stillbirth causes.It is recommended that each case of stillbirth placenta should undergo pathological examination.

Objective To establish the finite element model of spinal canal reconstruction and internal fixation,analysis influence of spinal canal reconstruction and internal fixation on spinal stability,and verify the effectiveness and reliability of spinal canal reconstruction and internal fixation in spinal canal surgery.Methods A 30-year-old male healthy volunteer with a height of 172 cm and weight of 75 kg was selected and his lumbar CT data were collected to establish a finite element model of normal lumbar Lo3-L,and the results were compared with in vitro solid results and published finite element analysis results to verify the validity of the model.They were divided into normal group,laminectomy group and spinal canal reconstruction group according to different treatment methods.Under the same boundary fixation and physiological load conditions,six kinds of ac-tivities were performed,including forward bending,backward extension,left bending,right bending,left rotation and right rota-tion,and the changes of range of motion(ROM)of L3-L4,L4-L5 segments and overall maximum ROM of L3-L5 were analyzed under the six conditions.Results The ROM displacement range of each segment of the constructed L3-L5 finite element model was consistent with the in vitro solid results and previous literature data,which confirms the validity of the model.In L3-L4,ROM of spinal canal reconstruction group was slightly increased than that of normal group during posterior extension(＞5％dif-ference),and ROM of other conditions was similar to that of normal group(＜5％difference).ROM in laminectomy group was significantly increase than that in normal group and spinal canal reconstruction group under the condition of flexion,extension,left and right rotation.In L4-L5,ROM in spinal canal reconstruction group was similar to that in normal group(＜5％differ-ence),while ROM in laminectomy group was significantly higher than that in normal group and spinal canal reconstruction group(＞5％difference).In the overall maximum ROM of L3-L5,spinal canal reconstruction group was only slightly higher than normal group under the condition of posterior extension(＞5％difference),while laminectomy was significantly higher than normal group and spinal canal reconstruction group under the condition of anterior flexion,posterior extension,left and right rotation(＞5％difference).The changes of each segment ROM and overall ROM of L3-L5 showed laminectomy group＞spinal canal reconstruction group＞normal group.Conclusion Laminectomy could seriously affect biomechanical stability of the spine,but application of spinal canal reconstruction and internal fixation could effectively reduce ROM displacement of the responsi-ble segment of spine and maintain its biomechanical stability.