ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ （AngⅡ） and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment， AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h， and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 （CCK-8） to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of B-type natriuretic peptide （BNP） in cell culture supernatant， and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently， the experiment was divided into a blank group （20% blank serum）， a model group （20% blank serum + 5×10^-5 mol·L^-1 AngⅡ）， low-， medium-， and high-dose （5%， 10% and 20%） serum containing Zhenwutang groups， an autophagy inhibitor group （1×10^-4 mol·L^-1 3-MA）， and autophagy inducer group （1×10^-7 mol·L^-1 rapamycin）. The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe （Lyso Tracker） and mitochondrial probe （Mito Tracker） co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect Caspase-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， and cytochrome C （Cyt C） in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase （Parkin）， phosphatase and tensin homolog （PTEN）-induced kinase 1 （PINK1）， and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3， cleaved Caspase-9， Bax， Bcl-2， and Cyt C in apoptosis-related pathways， phosphorylated ubiquitin ligase （p-Parkin）， phosphorylated PTEN-induced kinase 1 （p-PINK1）， p62， and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways， and the change of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10^-5 mol·L^-1， the cell activity was the lowest， and there was no cytotoxicity. At this concentration， the surface area of cardiomyocytes was significantly increased （P<0.01）， and the content of BNP in the supernatant of culture medium was significantly increased （P<0.05）. Therefore， AngⅡ with a concentration of 5×10^-5 mol·L^-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group， the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate （P<0.01） and significantly decreased mitochondrial membrane potential （P<0.01）. The results of immunofluorescence co-localization showed that compared with the blank group， the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group， the relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 in the model group was significantly up-regulated （P<0.01）， while the relative mRNA expression of Bcl-2， Parkin， and PINK1 was significantly down-regulated （P<0.01）. In addition， the relative protein expression of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 was significantly up-regulated （P<0.01）. The LC3Ⅱ/Ⅰ was significantly decreased， and the relative protein expression of Bcl-2， p-Parkin， p-PINK1， and Beclin1 was significantly down-regulated （P<0.01）. Compared with the model group， the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate （P<0.01）， and the decrease ratio of mitochondrial membrane potential is significantly lowered （P<0.01） in a dose-dependent manner. Additionally， both red and green fluorescence spots became more in these groups. In the 3-MA group， the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 was significantly down-regulated （P<0.05， P<0.01）， while that of Bcl-2， Parkin， and PINK1 was significantly up-regulated （P<0.01）. In the serum containing Zhenwutang groups， the relative protein expression levels of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 were significantly down-regulated （P<0.05，P<0.01）. The LC3Ⅱ/Ⅰ was significantly increased， and the relative protein expression levels of Bcl-2， p-Parkin， p-PINK1， and Beclin1 were significantly up-regulated （P<0.01）. ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

Definitive treatment of lung cancer with radiotherapy is challenging, as respiratory motion and anatomical changes can increase the risk of severe off-target effects during radiotherapy. Online adaptive radiotherapy (ART) is an evolving approach that enables timely modification of a treatment plan during the interfraction of radiotherapy, in response to physiologic or anatomic variations, aiming to improve the dose distribution for precise targeting and delivery in lung cancer patients. The effectiveness of online ART depends on the seamless integration of multiple components: sufficient quality of linear accelerator-integrated imaging guidance, deformable image registration, automatic recontouring, and efficient quality assurance and workflow. This review summarizes the present status of online ART for lung cancer, including key technologies, as well as the challenges and areas of active research in this field.
Humans ; Lung Neoplasms/radiotherapy* ; Radiotherapy Planning, Computer-Assisted/methods*

BACKGROUND: The available evidence regarding the benefits of percutaneous coronary intervention (PCI) on patients receiving dialysis with coronary artery disease (CAD) is limited and inconsistent. This study aimed to evaluate the association between PCI and clinical outcomes as compared with medical therapy alone in patients undergoing dialysis with CAD in China. METHODS: This multicenter, retrospective study was conducted in 30 tertiary medical centers across 12 provinces in China from January 2015 to June 2021 to include patients on dialysis with CAD. The primary outcome was major adverse cardiovascular events (MACE), defined as a composite of cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke. Secondary outcomes included all-cause death, the individual components of MACE, and Bleeding Academic Research Consortium criteria types 2, 3, or 5 bleeding. Multivariable Cox proportional hazard models were used to assess the association between PCI and outcomes. Inverse probability of treatment weighting (IPTW) and propensity score matching (PSM) were performed to account for potential between-group differences. RESULTS: Of the 1146 patients on dialysis with significant CAD, 821 (71.6%) underwent PCI. After a median follow-up of 23.0 months, PCI was associated with a 43.0% significantly lower risk for MACE (33.9% [ n  = 278] vs . 43.7% [ n  = 142]; adjusted hazards ratio 0.57, 95% confidence interval 0.45-0.71), along with a slightly increased risk for bleeding outcomes that did not reach statistical significance (11.1% vs . 8.3%; adjusted hazards ratio 1.31, 95% confidence interval, 0.82-2.11). Furthermore, PCI was associated with a significant reduction in all-cause and cardiovascular mortalities. Subgroup analysis did not modify the association of PCI with patient outcomes. These primary findings were consistent across IPTW, PSM, and competing risk analyses. CONCLUSION This study indicated that PCI in patients on dialysis with CAD was significantly associated with lower MACE and mortality when comparing with those with medical therapy alone, albeit with a slightly increased risk for bleeding events that did not reach statistical significance.
Humans ; Percutaneous Coronary Intervention/methods* ; Male ; Female ; Coronary Artery Disease/drug therapy* ; Retrospective Studies ; Renal Dialysis/methods* ; Middle Aged ; Aged ; China ; Proportional Hazards Models ; Treatment Outcome

OBJECTIVE: To explore mechanisms of acid-sensing ion channel 1 (ASIC1) mediated lumbar nucleus pulposus cell apoptosis through extracellular-signalregulated protein kinase 5 (ERK5) signaling pathway and mitochondrial dysfunction pathway. METHODS: Totally 34 patients with degenerative lumbar disc herniation (LDH) admitted from January 2020 to December 2022 were collected as research objects, including 21 males and 13 females;aged from 29 to 52 years old with an average of (37.43±4.75) years old;22 patients with grade Ⅱ and 12 patients with grade Ⅳ, according to Pfirrmann grading criteria;15 patients with L_4,5 and 19 patients with L₅S₁. The expression of ASIC1 in nucleus pulposus of LDH patients was measured by immunohistochemical staining. Nucleus pulposus cells were cultured by primary culture method, identified by toluidine blue staining and immunohistochemical staining, and the expression of ASIC1 protein was located by immunofluorescence staining. According to the addition of siRNA-ASIC1, ASIC1 overexpression plasmid, and ERK5 inhibitors, the nucleus pulpocyte was divided into three groups, named as SIRNA-silenced group, overexpression group, and inhibitor group, with 3 patients in each group. Cells of each group were collected at 72 h after intervention, expression of ASIC1, ERK5, BCL-xL/BCL-2-associated Death promoter (Bad), B-cell lymphoma-2 associated X (Bax) and B-cell lymphoblast-2 gene (Bcl-2) were detected by reverse transcription-polymerase chain reaction (RT-PCR);intracellular calcium ion levels were detected by calcium ion kit, mitochondrial membrane potential was detected by JC-1 kit, and apoptosis was observed by AV-PI kit. RESULTS: In LDH patients with grade Ⅳ, nucleus pulposus tissue removed during operation revealed poor elasticity, white color and poor ductility, and immunohistochemical results showed increased ASIC1 expression. There was no significant difference in mRNA relative expression of ASIC1 between siRNA silencing group (0.31±0.03) and inhibitor group (0.39±0.05) (P>0.05). The mRNA relative expression level of ERK5 in siRNA silencing group(0.32±0.05) was significantly higher than that in inhibitor group (0.15±0.04)(P<0.05), which suggested ERK5 was the downstream molecule of ASIC1. The mRNA relative expression levels of apoptosis promoting factor Bad and Bax in siRNA silencing group and inhibitor group were lower than those in overexpression group(P<0.05), the relative expression level of anti-apoptosis factor Bcl-2 mRNA was significantly increased (P<0.05). The calcium content in overexpression group was higher than that in siRNA silencing and inhibitor groups (P<0.05), the normal proportion of mitochondrial membrane potential in overexpression group was lower than that in siRNA silencing and inhibitor group (P<0.05), and the apoptosis rate in overexpression group was higher than that in siRNA silencing and inhibitor group (P<0.05). CONCLUSION After the activation of ASIC1 channel protein, calcium ions could enter the cells and act as a second messenger molecule to regulate apoptosis of nucleus pulposus cells by ERK5 signaling pathway and mitochondrial disorder pathway.
Humans ; Acid Sensing Ion Channels/physiology* ; Male ; Female ; Apoptosis ; Middle Aged ; Adult ; Signal Transduction ; Mitogen-Activated Protein Kinase 7/physiology* ; Mitochondrial Diseases/genetics* ; Nucleus Pulposus/metabolism* ; Intervertebral Disc Degeneration/metabolism* ; Mitochondria/metabolism* ; Intervertebral Disc Displacement/genetics*

Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

The structure of intestinal tissue is complex. In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases. Recently, organoids have been successfully constructed and they have come to play an important role in biomedical research. Organoids are miniaturized three-dimensional (3D) organs, derived from stem cells, which mimic the structure, cell types, and physiological functions of an organ, making them robust models for biomedical research. Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine, thereby providing a valuable platform for intestinal development and disease research. In this article, we review the latest progress in the construction and application of intestinal organoids.
Organoids/cytology* ; Intestines/physiology* ; Humans ; Animals ; Pluripotent Stem Cells

Objective:To investigate the pattern of distribution of the circumferential fascia of the rectum and elucidate its clinical implications.Methods:In this descriptive study, we examined the gross anatomy of four male hemipelvic cadaveric specimens from the Department of Anatomy at Fujian Medical University and the histological features of 16 fresh postoperative specimens from patients who had undergone total mesorectal excision for rectal cancer at the Department of Colorectal Surgery, Union Hospital, Fujian Medical University, between January and December 2022. The resultant combination of gross anatomical and histological features was employed to assess the following areas: (1)the morphology of the anterior mesorectum and fascia at the peritoneal reflection; (2)the caudal attachment point of Denonvilliers' fascia; (3) the fusion area of the pelvic plexus and the pre-hypogastric fascia; (4)the lateral and posterior attachment edges of the rectosacral fascia; and (5) selected histological features.Results:Our findings were as follows. (1) At the peritoneal reflection, the anterior mesorectum forms a triangular fat pad with a dense fascial structure. The base of this pad extends anteriorly across the most caudal point of the peritoneal reflection, with Denonvilliers' fascia originating from the anterior side of the triangle, near the bladder side of the peritoneum craniad to the peritoneal reflection. (2) The caudal attachment of Denonvilliers' fascia is at the angle between the seminal vesicles, the ampulla of the vas deferens, and the prostate. It adheres tightly to the prostatic capsule and vascular bundles pass through its cephalic side. (3) The pre-hypogastric fascia transitions laterally to merge with Denonvilliers' fascia; its middle part being inseparable from the main body of the pelvic plexus, which gives rise to the nerves that innervate the rectum. (4) The rectosacral fascia is formed by fusion of the fascia propria with the pre-hypogastric fascia. The resultant fused fascia bifurcates into two leaves on the right side; the outer leaf being the pre-hypogastric fascia and the inner leaf the fascia propria. (5) Histologically, the peritoneal reflection zone shows cuboidal epithelium of the peritoneum at its lowest point with no detectable origin of Denonvilliers' fascia. The anterior side of the peritoneal reflection, from which Denonvilliers' fascia originates, has a dense double-layered fascial structure comprising thick collagen fiber (16/16). The fascia propria exhibits a thinner and looser collagen fiber structure and its origin varies between individuals, 13/16 originating together with Denonvilliers' fascia from the craniad side of the peritoneal reflection, and 3/16 originating separately from the most caudal point of the peritoneal reflection. The caudal edge of Denonvilliers' fascia has a double-layered fascial structure with multiple S100-stained areas. The posterior edge of the rectosacral fascia has a fused fascial structure, thick nerve fibers being clearly observable between collagen fibers originating from the pre-hypogastric fascia under high magnification. The lateral edge of the rectosacral fascia extends interiorly, maintains the integrity of the fascia propria.Conclusions:In this study, we investigated the pattern of distribution of the circumferential fascia of the rectum by cadaveric dissection and histological examination of postoperative specimens. We found that the anterior mesorectum forms a triangular fat pad that can serve as a reference for dissection anterior to Denonvilliers' fascia, by making incisions 1 cm above the peritoneal reflection. The region of fusion of Denonvilliers' fascia with the prostatic capsule on the caudal side is rich in neurovascular bundles, contradicting the traditional view of a retroprostatic plane. This finding supports the practice of cutting Denonvilliers' fascia 0.5 cm above the base of the seminal vesicles. The fusion of the fascia propria with the pre-hypogastric fascia posteriorly forms the rectosacral fascia, which bifurcates into two leaves on both sides of the rectum, the inner leaf being the fascia propria and the outer leaf the pre-hypogastric fascia. These transition anteriorly to become Denonvilliers' fascia and fuse densely with the main body of the pelvic plexus on both sides. These findings provide a theoretical foundation for protecting the pelvic plexus and hypogastric nerve by transecting Denonvilliers' fascia and then dissecting in a top-to-bottom direction (i.e., from anterior to caudal), ultimately leading to the transection of the pre-hypogastric fascia.

Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.

Objective:To investigate the pattern of distribution of the circumferential fascia of the rectum and elucidate its clinical implications.Methods:In this descriptive study, we examined the gross anatomy of four male hemipelvic cadaveric specimens from the Department of Anatomy at Fujian Medical University and the histological features of 16 fresh postoperative specimens from patients who had undergone total mesorectal excision for rectal cancer at the Department of Colorectal Surgery, Union Hospital, Fujian Medical University, between January and December 2022. The resultant combination of gross anatomical and histological features was employed to assess the following areas: (1)the morphology of the anterior mesorectum and fascia at the peritoneal reflection; (2)the caudal attachment point of Denonvilliers' fascia; (3) the fusion area of the pelvic plexus and the pre-hypogastric fascia; (4)the lateral and posterior attachment edges of the rectosacral fascia; and (5) selected histological features.Results:Our findings were as follows. (1) At the peritoneal reflection, the anterior mesorectum forms a triangular fat pad with a dense fascial structure. The base of this pad extends anteriorly across the most caudal point of the peritoneal reflection, with Denonvilliers' fascia originating from the anterior side of the triangle, near the bladder side of the peritoneum craniad to the peritoneal reflection. (2) The caudal attachment of Denonvilliers' fascia is at the angle between the seminal vesicles, the ampulla of the vas deferens, and the prostate. It adheres tightly to the prostatic capsule and vascular bundles pass through its cephalic side. (3) The pre-hypogastric fascia transitions laterally to merge with Denonvilliers' fascia; its middle part being inseparable from the main body of the pelvic plexus, which gives rise to the nerves that innervate the rectum. (4) The rectosacral fascia is formed by fusion of the fascia propria with the pre-hypogastric fascia. The resultant fused fascia bifurcates into two leaves on the right side; the outer leaf being the pre-hypogastric fascia and the inner leaf the fascia propria. (5) Histologically, the peritoneal reflection zone shows cuboidal epithelium of the peritoneum at its lowest point with no detectable origin of Denonvilliers' fascia. The anterior side of the peritoneal reflection, from which Denonvilliers' fascia originates, has a dense double-layered fascial structure comprising thick collagen fiber (16/16). The fascia propria exhibits a thinner and looser collagen fiber structure and its origin varies between individuals, 13/16 originating together with Denonvilliers' fascia from the craniad side of the peritoneal reflection, and 3/16 originating separately from the most caudal point of the peritoneal reflection. The caudal edge of Denonvilliers' fascia has a double-layered fascial structure with multiple S100-stained areas. The posterior edge of the rectosacral fascia has a fused fascial structure, thick nerve fibers being clearly observable between collagen fibers originating from the pre-hypogastric fascia under high magnification. The lateral edge of the rectosacral fascia extends interiorly, maintains the integrity of the fascia propria.Conclusions:In this study, we investigated the pattern of distribution of the circumferential fascia of the rectum by cadaveric dissection and histological examination of postoperative specimens. We found that the anterior mesorectum forms a triangular fat pad that can serve as a reference for dissection anterior to Denonvilliers' fascia, by making incisions 1 cm above the peritoneal reflection. The region of fusion of Denonvilliers' fascia with the prostatic capsule on the caudal side is rich in neurovascular bundles, contradicting the traditional view of a retroprostatic plane. This finding supports the practice of cutting Denonvilliers' fascia 0.5 cm above the base of the seminal vesicles. The fusion of the fascia propria with the pre-hypogastric fascia posteriorly forms the rectosacral fascia, which bifurcates into two leaves on both sides of the rectum, the inner leaf being the fascia propria and the outer leaf the pre-hypogastric fascia. These transition anteriorly to become Denonvilliers' fascia and fuse densely with the main body of the pelvic plexus on both sides. These findings provide a theoretical foundation for protecting the pelvic plexus and hypogastric nerve by transecting Denonvilliers' fascia and then dissecting in a top-to-bottom direction (i.e., from anterior to caudal), ultimately leading to the transection of the pre-hypogastric fascia.