Incomplete histologic healing and diminished biomechanical strength of meniscus-bone interface after medial meniscus posterior root transosseous repair in a goat model.

PURPOSE: To enhance the understanding of histological healing after repairing medial meniscal posterior root tear (MMPRT) at an early stage, utilizing a goat model. METHODS: Eighteen adult goats, totaling thirty-six knee joints, were allocated into three groups (n = 12): Sham group (Sham), Root Tear group (RT), and Root Tear with Transosseous Suture group (RTS). At 12- and 24-week intervals post-surgery, all the knees were harvested for imaging, macroscopic, histological, and biomechanical assessments. RESULTS: The intact root served as a meniscus-bone interface which connected the tibial and the circular fibers of the meniscus, with a bony insertion and a root-meniscus transition. A direct-fibrous-connection displayed at the bony insertion proximal to the synovium in the RTS group, while the remaining regions of the root displayed indirect-fibrous healing. The healing in the RT group was disjointed and reminiscent of scar tissue. The RTS group exhibited a more pronounced coronal extrusion compared to the Sham group (0.42 ± 0.09 vs. 0.19 ± 0.02, P = 0.0012) but was improved relative to that of the RT group (0.49 ± 0.02, P = 0.0028). The failure load and stiffness of the RTS group were notably higher than those of the RT group, with a strength of 42.67% and a stiffness of 83.75% of the intact root. All the samples ruptured at the root-meniscus transitions. CONCLUSION: The incomplete healing may be attributed to the histological factors underlying the low healing rate and persistent MME. Notably, the region attached to the posterior-cruciate-ligament exhibited superior healing compared to other regions of the bony insertion in the repaired group. Conversely, the root-meniscus transition displayed discontinuity, representing a mechanical weakness in the healing process. CLINICAL RELEVANCE: Modifications of bone tunnel positioning and suture placement could be undertaken in subsequent studies to particularly enhance the healing of the root-meniscus transition.

Air disinfection performance of upper-room ultraviolet germicidal irradiation (UR-UVGI) system in a multi-compartment dental clinic.

Multi-compartment dental clinics present significant airborne cross-infection risks. Upper-room ultraviolet germicidal irradiation (UR-UVGI) system have shown promise in preventing airborne pathogens, but its available application data are insufficient in multi-compartment dental clinics. Therefore, the UR-UVGI system's performance in a multi-compartment dental clinic was comprehensively evaluated in this study. The accuracy of the turbulence and drift flux models was verified by experimental data from ultrasonic scaling. The effects of the ventilation rate, irradiation zone volume, and irradiation flux on UR-UVGI performance were analyzed using computational fluid dynamics coupled with a UV inactivation model. Different patient numbers were considered. The results showed that UR-UVGI significantly reduced virus concentrations and outperformed increased ventilation rates alone. At a ventilation rate of six air changes per hour (ACH), UR-UVGI with an irradiation zone volume of 20% and irradiation flux of 5 µW/cm2 achieved a 70.44% average virus reduction in the whole room (WR), outperforming the impact of doubling the ventilation rate from 6 to 12 ACH without UR-UVGI. The highest disinfection efficiency of UR-UVGI decreased for WRs with more patients. The compartment treating patients exhibited significantly lower disinfection efficiency than others. Moreover, optimal UR-UVGI performance occurs at lower ventilation rates, achieving over 80% virus disinfection in WR. Additionally, exceeding an irradiation zone volume of 20% or an irradiation flux of 5 µW/cm2 notably reduces the improvement rates of UR-UVGI performance. These findings provide a scientific reference for strategically applying UR-UVGI in multi-compartment dental clinics.

Evaluation of the extended Japan NBI expert team classification of subtype 2B in laterally spreading colorectal tumors based on blue laser imaging.

OBJECTIVES: The Japan NBI Expert Team (JNET) classification has good diagnostic potential for colorectal diseases. We aimed to explore the diagnostic value of the JNET classification type 2B (JNET2B) criteria for colorectal laterally spreading tumors (LSTs) based on magnifying endoscopy with blue laser imaging (ME-BLI) examination. METHODS: Between January 2017 and June 2023, 218 patients who were diagnosed as having JNET2B-type LSTs using ME-BLI were included retrospectively. Endoscopic images were reinterpreted to categorize the LSTs as JNET2B-low (n = 178) and JNET2B-high (n = 53) LSTs. The JNET2B-low and JNET2B-high LSTs were compared based on their histopathological and morphological classifications. RESULTS: Among the 178 JNET2B-low LSTs, 86 (48.3%) were histopathologically classified as low-grade intraepithelial neoplasia, 54 (30.3%) as high-grade intraepithelial neoplasia (HGIN), 37 (20.8%) as intramucosal carcinoma (IMC), and one (0.6%) as superficial invasive submucosal carcinoma (SMC1). Among the 53 JNET2B-high LSTs, five (9.4%) were classified as HGIN, 28 (52.9%) as IMC, 15 (28.3%) as SMC1, and 5 (9.4%) as deep invasive submucosal carcinoma. There were significant differences in this histopathological classification between the two groups (P < 0.001). However, there was no significant difference between JNET2B-low and JNET2B-high LSTs based on their morphological classification (granular vs nongranular) or size (<20 mm vs ≥20 mm). Besides, the κ value for JNET2B subtyping was 0.698 (95% confidence interval 0.592-0.804) between the two endoscopists who reassessed the endoscopic images. CONCLUSION: The JNET2B subtyping of LSTs has a diagnostic potential in the preoperative setting, and may be valuable for treatment decision-making.

Deficiency of IQCH causes male infertility in humans and mice.

IQ motif-containing proteins can be recognized by calmodulin (CaM) and are essential for many biological processes. However, the role of IQ motif-containing proteins in spermatogenesis is largely unknown. In this study, we identified a loss-of-function mutation in the novel gene IQ motif-containing H (IQCH) in a Chinese family with male infertility characterized by a cracked flagellar axoneme and abnormal mitochondrial structure. To verify the function of IQCH, Iqch knockout (KO) mice were generated via CRISPR-Cas9 technology. As expected, the Iqch KO male mice exhibited impaired fertility, which was related to deficient acrosome activity and abnormal structures of the axoneme and mitochondria, mirroring the patient phenotypes. Mechanistically, IQCH can bind to CaM and subsequently regulate the expression of RNA-binding proteins (especially HNRPAB), which are indispensable for spermatogenesis. Overall, this study revealed the function of IQCH, expanded the role of IQ motif-containing proteins in reproductive processes, and provided important guidance for genetic counseling and genetic diagnosis of male infertility.

Comparative study of the effects of custom-made insole and ordinary insole in adults with flexible flatfoot on different slopes.

BACKGROUND: Flatfoot (pes planus) is a common foot deformity, and its causes are mainly related to age, gender, weight, and genetics. Previous studies have shown that custom-made insoles could have a positive effect in improving plantar pressure and symptoms in individuals with flexible flatfeet, but it remains to be explored whether they can still show benefits in daily walking on different slopes. OBJECTIVE: This study aims to investigate a custom-made insole based on plantar pressure redistribution and to verify its effectiveness by gait analysis on different slopes. METHODS: We recruited 10 subjects and compared the peak pressure and impulse in each area between custom-made insole (CI) and ordinary insole (OI) groups. RESULTS: The results illustrate that CI raises the pressure in T area, improves the ability of the subjects to move forward in the slope walking, which was beneficial to gait stability. CONCLUSION: The redistribution of pressure in MF and MH area is promoted to provide active protection for subjects. Meanwhile, CI could decrease the impulse in MF area during uphill and level walking, which effectively reduces the accumulation of fatigue during gait. Moreover, avoiding downhill walking could be able to protect foot from injury in daily life.

Selected Lark Mitochondrial Genomes Provide Insights into the Evolution of Second Control Region with Tandem Repeats in Alaudidae (Aves, Passeriformes).

The control region (CR) regulates the replication and transcription of the mitochondrial genome (mitogenome). Some avian mitogenomes possess two CRs, and the second control region (CR2) may enhance replication and transcription; however, the CR2 in lark mitogenome appears to be undergoing loss and is accompanied by tandem repeats. Here, we characterized six lark mitogenomes from Alaudala cheleensis, Eremophila alpestris, Alauda razae, and Calandrella cinerea and reconstructed the phylogeny of Passerida. Through further comparative analysis among larks, we traced the evolutionary process of CR2. The mitochondrial gene orders were conserved in all published lark mitogenomes, with Cytb-trnT-CR1-trnP-ND6-trnE-remnant CR2 with tandem repeat-trnF-rrnS. Phylogenetic analysis revealed Alaudidae and Panuridae are sister groups at the base of Sylvioidea, and sporadic losses of CR2 may occur in their common ancestor. CR sequence and phylogeny analysis indicated CR2 tandem repeats were generated within CR2, originating in the ancestor of all larks, rather than inherited from CR1. The secondary structure comparison of tandem repeat units within and between species suggested slipped-strand mispairing and DNA turnover as suitable models for explaining the origin and evolution of these repeats. This study reveals the evolutionary process of the CR2 containing tandem repeat in Alaudidae, providing reference for understanding the evolutionary characteristics and dynamics of tandem repeats.

HSF5 Deficiency Causes Male Infertility Involving Spermatogenic Arrest at Meiotic Prophase I in Humans and Mice.

Meiosis is a specialized cell division process that generates gametes for sexual reproduction. However, the factors and underlying mechanisms involving meiotic progression remain largely unknown, especially in humans. Here, it is first showed that HSF5 is associated with human spermatogenesis. Patients with a pathogenic variant of HSF5 are completely infertile. Testicular histologic findings in the patients reveal rare postmeiotic germ cells resulting from meiotic prophase I arrest. Hsf5 knockout (KO) mice confirms that the loss of HSF5 causes defects in meiotic recombination, crossover formation, sex chromosome synapsis, and sex chromosome inactivation (MSCI), which may contribute to spermatocyte arrest at the late pachytene stage. Importantly, spermatogenic arrest can be rescued by compensatory HSF5 adeno-associated virus injection into KO mouse testes. Mechanistically, integrated analysis of RNA sequencing and chromatin immunoprecipitation sequencing data revealed that HSF5 predominantly binds to promoters of key genes involved in crossover formation (e.g., HFM1, MSH5 and MLH3), synapsis (e.g., SYCP1, SYCP2 and SYCE3), recombination (TEX15), and MSCI (MDC1) and further regulates their transcription during meiotic progression. Taken together, the study demonstrates that HSF5 modulates the transcriptome to ensure meiotic progression in humans and mice. These findings will aid in genetic diagnosis of and potential treatments for male infertility.

Spatial distribution of bioaerosols and evaluation of four ventilation method on controlling their diffusion in a typical enhanced biosafety level 2 laboratory.

Biosafety laboratories are critical in many fields. However, experimenters associated the infection risk from biological aerosols. In this study, by conducting experiments on the release and collection of bioaerosols within a typical BSL-2 + laboratory, the spatial distribution of bioaerosols was tracked. Numerical calculations were employed to obtain and visualize the airflow patterns and aerosol dispersion paths of four ventilation methods. The results indicated that equipment and tables led to uneven airflow distribution within the laboratory. The comparison results of the four evaluation indicators showed that the air age distribution of UU (Upward supply and upward return) mode and CD (Cross-supply and downward return) mode was superior, with air change efficiency values of 0.595 and 0.603, respectively. Additionally, the contaminant removal index of CD mode was 1.48, significantly higher than the other ventilation methods. The statistical results of the contaminant dispersion index also indicated that CD mode was most conducive to diluting aerosols in the spatial environment. The LD (lateral supply and downward return) mode may lead to airflow short-circuiting. The UD (upward supply and downward return) mode can provide balanced protection for laboratory. Overall, CD mode performed the best among the four ventilation methods, followed by UU mode.

Ferroelectric field enhanced tribocatalytic hydrogen production and RhB dye degradation by tungsten bronze ferroelectrics.

Tribocatalysis is a method that converts mechanical energy into chemical energy. In this study, we synthesized tungsten bronze structured Ba0.75Sr0.25Nb1.9Ta0.1O6 ferroelectric ceramic submicron powder using a traditional solid-state route, and the powder exhibited excellent performance in tribocatalytic water splitting for hydrogen production. Under the friction stirring of three polytetrafluoroethylene (PTFE) magnetic stirring bars in pure water, the rate of hydrogen generation by the Ba0.75Sr0.25Nb1.9Ta0.1O6 ferroelectric submicron powder is 200 µmol h-1 g-1, and after 72 hours, the accumulated hydrogen production reaches 15 892.8 µmol g-1. Additionally, this ferroelectric tungsten bronze ferroelectric material also exhibits excellent tribocatalytic degradation ability toward RhB dyes, with degradation efficiency reaching 96% in 2 hours. The study of tribocatalysis based on tungsten bronze ferroelectric materials represents a significant step forward in versatile energy utilization for clean energy and environmental wastewater degradation.

Spatially ordered NiOOH-ZnS/CdS heterostructures with an efficient photo-carrier transmission channel for markedly improved H2 production.

Spatially-ordered 1D nanocrystal-based semiconductor nanostructures possess distinct merits for photocatalytic reaction, including large surface area, fast carrier separation, and enhanced light scattering and absorption. Nevertheless, establishing a valid photo-carrier transmission channel is still crucial yet challenging for semiconductor heterostructures to realize efficient photocatalysis. In this work, spatially ordered NiOOH-ZnS/CdS heterostructures were constructed by sequential ZnS coating and NiOOH photo-deposition on multi-armed CdS, which consists of {112̄0}-faceted wurtzite nanorods grown epitaxially on {111}-faceted zinc blende core. Intriguingly, the surface photovoltage spectroscopy and PbO2 photo-deposition results suggest that the photogenerated holes of CdS were first transferred to the Zn-vacancy level of ZnS and then to NiOOH, as driven by the built-in electric field between ZnS and CdS and the hole-extracting effect of the NiOOH cocatalyst, leading to the efficient charge separation of NiOOH-ZnS/CdS. With visible-light (λ > 420 nm) irradiation, NiOOH-ZnS/CdS exhibited a distinguished H2-evolution rate of 152.20 mmol g-1 h-1 (apparent quantum efficiency of 40.9% at 420 nm), approximately 18 folds that of 3 wt% Pt-loaded CdS and much higher than that of ZnS/CdS and NiOOH-CdS counterparts as well as the most reported CdS-containing photocatalysts. Moreover, the cycling and long-term H2 generation tests manifested the outstanding photocatalyst stability of NiOOH-ZnS/CdS. The study results presented here may propel the controllable design of highly-active nanomaterials for solar conversion and utilization.

KLF13 promotes VSMCs phenotypic dedifferentiation by directly binding to the SM22α promoter.

Krüppel-like factor 13 (KLF13), a zinc finger transcription factor, is considered as a potential regulator of cardiomyocyte differentiation and proliferation during heart morphogenesis. However, its precise role in the dedifferentiation of vascular smooth muscle cells (VSMCs) during atherosclerosis and neointimal formation after injury remains poorly understood. In this study, we investigated the relationship between KLF13 and SM22α expression in normal and atherosclerotic plaques by bioanalysis, and observed a significant increase in KLF13 levels in the atherosclerotic plaques of both human patients and ApoE-/- mice. Knockdown of KLF13 was found to ameliorate intimal hyperplasia following carotid artery injury. Furthermore, we discovered that KLF13 directly binds to the SM22α promoter, leading to the phenotypic dedifferentiation of VSMCs. Remarkably, we observed a significant inhibition of platelet-derived growth factor BB-induced VSMCs dedifferentiation, proliferation, and migration when knocked down KLF13 in VSMCs. This inhibitory effect of KLF13 knockdown on VCMC function was, at least in part, mediated by the inactivation of p-AKT signaling in VSMCs. Overall, our findings shed light on a potential therapeutic target for treating atherosclerotic lesions and restenosis after vascular injury.

COMP promotes pancreatic fibrosis by activating pancreatic stellate cells through CD36-ERK/AKT signaling pathways.

BACKGROUND: Pancreatic fibrosis is one of the most important pathological features of chronic pancreatitis (CP) and pancreatic stellate cells (PSCs) are the key cells of fibrosis. As an extracellular matrix (ECM) glycoprotein, cartilage oligomeric matrix protein (COMP) is critical for collagen assembly and ECM stability and recent studies showed that COMP exert promoting fibrosis effect in the skin, lungs and liver. However, the role of COMP in activation of PSCs and pancreatic fibrosis remain unclear. We aimed to investigate the role and specific mechanisms of COMP in regulating the profibrotic phenotype of PSCs and pancreatic fibrosis. METHODS: ELISA method was used to determine serum COMP in patients with CP. Mice model of CP was established by repeated intraperitoneal injection of cerulein and pancreatic fibrosis was evaluated by Hematoxylin-Eosin staining (H&E) and Sirius red staining. Immunohistochemical staining was used to detect the expression changes of COMP and fibrosis marker such as α-SMA and Fibronectin in pancreatic tissue of mice. Cell Counting Kit-8, Wound Healing and Transwell assessed the proliferation and migration of human pancreatic stellate cells (HPSCs). Western blotting, qRT-PCR and immunofluorescence staining were performed to detect the expression of fibrosis marker, AKT and MAPK family proteins in HPSCs. RNA-seq omics analysis as well as small interfering RNA of COMP, recombinant human COMP (rCOMP), MEK inhibitors and PI3K inhibitors were used to study the effect and mechanism of COMP on activation of HPSCs. RESULTS: ELISA showed that the expression of COMP significantly increased in the serum of CP patients. H&E and Sirius red staining analysis showed that there was a large amount of collagen deposition in the mice in the CP model group and high expression of COMP, α-SMA, Fibronectin and Vimentin were observed in fibrotic tissues. TGF-ß1 stimulates the activation of HPSCs and increases the expression of COMP. Knockdown of COMP inhibited proliferation and migration of HPSCs. Further, RNA-seq omics analysis and validation experiments in vitro showed that rCOMP could significantly promote the proliferation and activation of HPSCs, which may be due to promoting the phosphorylation of ERK and AKT through membrane protein receptor CD36. rCOMP simultaneously increased the expression of α-SMA, Fibronectin and Collagen I in HPSCs. CONCLUSION: In conclusion, this study showed that COMP was up-regulated in CP fibrotic tissues and COMP induced the activation, proliferation and migration of PSCs through the CD36-ERK/AKT signaling pathway. COMP may be a potential therapeutic candidate for the treatment of CP. Interfering with the expression of COMP or the communication between COMP and CD36 on PSCs may be the next direction for therapeutic research.

NEK2 affects the ferroptosis sensitivity of gastric cancer cells by regulating the expression of HMOX1 through Keap1/Nrf2.

NEK2 is a serine/threonine protein kinase that is involved in regulating the progression of various tumors. Our previous studies have found that NEK2 is highly expressed in gastric cancer and suggests that patients have a worse prognosis. However, its role and mechanism in gastric cancer are only poorly studied. In this study, we established a model of ferroptosis induced by RSL3 or Erastin in AGS cells in vitro, and konckdown NEK2, HOMX1, Nrf2 by siRNA. The assay kit was used to analyzed cell viability, MDA levels, GSH and GSSG content, and FeRhoNox™-1 fluorescent probe, BODIPY™ 581/591 C11 lipid oxidation probe, CM-H2DCFDA fluorescent probe were used to detected intracellular Fe2+, lipid peroxidation, and ROS levels, respectively. Calcein-AM/PI staining was used to detect the ratio of live and dead cells, qRT-PCR and Western blot were used to identify the mRNA and protein levels of genes in cells, immunofluorescence staining was used to analyze the localization of Nrf2 in cells, RNA-seq was used to analyze changes in mRNA expression profile, and combined with the FerrDb database, ferroptosis-related molecules were screened to elucidate the impact of NEK2 on the sensitivity of gastric cancer cells to ferroptosis. We found that inhibition of NEK2 could enhance the sensitivity of gastric cancer cells to RSL3 and Erastin-induced ferroptosis, which was reflected in the combination of inhibition of NEK2 and ferroptosis induction compared with ferroptosis induction alone: cell viability and GSH level were further decreased, while the proportion of dead cells, Fe2+ level, ROS level, lipid oxidation level, MDA level, GSSG level and GSSG/GSH ratio were further increased. Mechanism studies have found that inhibiting NEK2 could promote the expression of HMOX1, a gene related to ferroptosis, and enhance the sensitivity of gastric cancer cells to ferroptosis by increasing HMOX1. Further mechanism studies have found that inhibiting NEK2 could promote the ubiquitination and proteasome degradation of Keap1, increase the level of Nrf2 in the nucleus, and thus promote the expression of HMOX1. This study confirmed that NEK2 can regulate HMOX1 expression through Keap1/Nrf2 signal, and then affect the sensitivity of gastric cancer cells to ferroptosis, enriching the role and mechanism of NEK2 in gastric cancer.

Comparative analysis of bone turnover markers in bone marrow and peripheral blood: implications for osteoporosis.

INTRODUCTION: This study examines bone turnover marker (BTM) variations between bone marrow and peripheral blood in osteoporotic and non-osteoporotic patients. BTMs offer insights into bone remodeling, crucial for understanding osteoporosis. METHODS: A total of 133 patients were categorized into osteoporotic and non-osteoporotic cohorts. BTMs-C-telopeptide cross-linked type 1 collagen (ß-CTX), serum osteocalcin (OC), Procollagen type I N-propeptide (P1NP), 25(OH)D-were measured in bone marrow and peripheral blood. Lumbar spine bone mineral density (BMD) was assessed. RESULTS: Osteoporotic patients exhibited elevated ß-CTX and OC levels in peripheral blood, indicating heightened bone resorption and turnover. ß-CTX levels in osteoporotic bone marrow were significantly higher. Negative correlations were found between peripheral blood ß-CTX and OC levels and lumbar spine BMD, suggesting their potential as osteoporosis severity indicators. No such correlations were observed with bone marrow markers. When analyzing postmenopausal women separately, we obtained consistent results. CONCLUSIONS: Elevated ß-CTX and OC levels in osteoporotic peripheral blood highlight their diagnostic significance. Negative ß-CTX and OC-BMD correlations underscore their potential for assessing osteoporosis severity. Discrepancies between peripheral blood and bone marrow markers emphasize the need for further exploration. This research advances our understanding of BTM clinical applications in osteoporosis diagnosis and treatment.

Steep Posterior Tibial Slope and Excessive Anterior Tibial Translation Are Associated With Increased Sagittal Meniscal Extrusion After Posterior Lateral Meniscus Root Repair Combined With Anterior Cruciate Ligament Reconstruction.

Purpose: To (1) evaluate the clinical and radiographic outcomes of patients with primary anterior cruciate ligament reconstruction (ACLR) with type II posterior lateral meniscus root tear (PLMRT) repair and (2) identify whether increased anterior tibial subluxation of the lateral compartment (ATSLC) and steeper posterior tibial slope (PTS) are associated with sagittal lateral meniscal extrusion (LME). Methods: Patients who underwent primary anatomic ACLR with concomitant type II PLMRTs using the all-inside side-to-side repair technique between November 2014 and September 2020 were identified. To be included, patients must have had a minimum of 2 years follow-up. All patients, including those with ATSLC and PTS and sagittal and coronal LME, were retrospectively reviewed clinically and radiologically. The patients were divided into 2 subgroups according to the occurrence of sagittal LME. Results: Forty patients were included in this study with a mean follow-up of 44 months (range, 24-94 months). In general, the postoperative parameters, including grade of pivot shift, side-to-side difference, ATSLC, Lysholm score, and International Knee Documentation Committee (IKDC) score, were significantly improved compared with the preoperative ones. However, postoperative sagittal LME was detected to be significantly larger than the preoperative one. Minimal clinically important difference (MCID) analysis for postoperative outcomes showed that the rate of patients who achieved MCID thresholds was 100% for Lysholm, 95% for IKDC, 42.50% for coronal LME, 62.50% for sagittal LME, 40% for ATSLC, and 100% for side-to-side difference. Further comparisons, where patients were divided into 2 subgroups according to the occurrence of sagittal LME, showed significant differences in PTS, ATSLC, and coronal LME. Conclusions: Clinical outcomes after type II PLMRT repair with primary ACLR were significantly improved, except for LME, at the 2-year postoperative follow-up. After repair of type II PLMRT injuries, the presence of sagittal LME was associated with increased PTS and ATSLC. Level of Evidence: Level III, retrospective cohort study.

Artificial intelligence-assisted system for the assessment of Forrest classification of peptic ulcer bleeding: a multicenter diagnostic study.

BACKGROUND: Inaccurate Forrest classification may significantly affect clinical outcomes, especially in high risk patients. Therefore, this study aimed to develop a real-time deep convolutional neural network (DCNN) system to assess the Forrest classification of peptic ulcer bleeding (PUB). METHODS: A training dataset (3868 endoscopic images) and an internal validation dataset (834 images) were retrospectively collected from the 900th Hospital, Fuzhou, China. In addition, 521 images collected from four other hospitals were used for external validation. Finally, 46 endoscopic videos were prospectively collected to assess the real-time diagnostic performance of the DCNN system, whose diagnostic performance was also prospectively compared with that of three senior and three junior endoscopists. RESULTS: The DCNN system had a satisfactory diagnostic performance in the assessment of Forrest classification, with an accuracy of 91.2% (95%CI 89.5%-92.6%) and a macro-average area under the receiver operating characteristic curve of 0.80 in the validation dataset. Moreover, the DCNN system could judge suspicious regions automatically using Forrest classification in real-time videos, with an accuracy of 92.0% (95%CI 80.8%-97.8%). The DCNN system showed more accurate and stable diagnostic performance than endoscopists in the prospective clinical comparison test. This system helped to slightly improve the diagnostic performance of senior endoscopists and considerably enhance that of junior endoscopists. CONCLUSION: The DCNN system for the assessment of the Forrest classification of PUB showed satisfactory diagnostic performance, which was slightly superior to that of senior endoscopists. It could therefore effectively assist junior endoscopists in making such diagnoses during gastroscopy.

Novel variations in TENT5D lead to teratozoospermia in infertile patients.

PURPOSE: Teratozoospermia is the main pathogenic factor of male infertility. However, the genetic etiology of teratozoospermia is largely unknown. This study aims to clarify the relationship between novel variations in TENT5D and teratozoospermia in infertile patients. MATERIALS AND METHODS: Two infertile patients were enrolled. Routine semen analysis of patients and normal controls was conducted with the WHO guidelines. Whole-exome sequencing (WES) was conducted to identify pathogenic variants in the two patients. Morphology and ultrastructure analysis of spermatozoa in the two patients was determined by Papanicolaou staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The functional effect of the identified variants was analyzed by immunofluorescence staining and western blotting. The expression of TENT5D in different germ cells was detected by immunofluorescence staining. RESULTS: Two new hemizygous variations, c.101C > T (p.P34L) and c.125A > T (p.D42V), in TENT5D were detected in two patients with male infertility. Morphology analysis showed abnormalities in spermatozoa morphology in the two patients, including multiple heads, headless, multiple tails, coiled, and/or bent flagella. Ultrastructure analysis showed that most of the spermatozoa exhibited missing or irregularly arranged '9+2' structures. Further functional experiments confirmed the abrogated TENT5D protein expression in patients. In addition, both p.P34L and p.D42V substitutions resulted in a conformational change of the TENT5D protein. We precisely analyzed the subcellular localization of TENT5D in germ cells in humans and mice. And we found that TENT5D was predominantly detected in the head and flagellum of elongating spermatids and epididymal spermatozoa. CONCLUSIONS: Our results showed further evidence of a relationship between TENT5D mutation and human male infertility, providing new genetic insight for use in the diagnosis and treatment of male infertility.

A novel SNP in HUWE1 promoter confers increased risk of NOA by affecting the RA/RARα pathway in Chinese individuals.

BACKGROUND: The ubiquitin ligase HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 is essential for the establishment and maintenance of spermatogonia. However, the role of HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 in regulating germ cell differentiation remains unclear, and clinical evidence linking HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 to male infertility pathogenesis is lacking. OBJECTIVE: This study aims to investigate the role of HUWE1 in germ cell differentiation and the mechanism by which a HUWE1 single nucleotide polymorphism increases male infertility risk. MATERIALS AND METHODS: We analyzed HUWE1 single nucleotide polymorphisms in 190 non-obstructive azoospermia patients of Han Chinese descent. We evaluated HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 regulation by retinoic acid receptor alpha using chromatin immunoprecipitation assays, electrophoretic mobility shift assays, and siRNA-mediated RARα knockdown. Using C18-4 spermatogonial cells, we determined whether HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 participated in retinoic acid-mediated retinoic acid receptor alpha signaling. We performed luciferase assays, cell counting kit-8 assays, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting. We quantified HUWE1 and retinoic acid receptor alpha in testicular biopsies from non-obstructive azoospermia and obstructive azoospermia patients using quantitative real-time polymerase chain reaction and immunofluorescence. RESULTS: Three HUWE1 single nucleotide polymorphisms were significantly associated with spermatogenic failure in 190 non-obstructive azoospermia patients; one (rs34492591) was in the HUWE1 promoter. Retinoic acid receptor alpha regulates HUWE1 gene expression by binding to its promoter. HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 participates in retinoic acid/retinoic acid receptor alpha signaling pathway and regulates the expression of germ cell differentiation genes STRA8 and SCP3 to inhibit cell proliferation and reduce γH2AX accumulation. Notably, significantly lower levels of HUWE1 and RARα were detected in testicular biopsy samples from non-obstructive azoospermia patients. CONCLUSIONS: An HUWE1 promoter single nucleotide polymorphism significantly downregulates its expression in non-obstructive azoospermia patients. Mechanistically, HECT, UBA, and WWE domain-containing E3 ubiquitin protein ligase 1 regulates germ cell differentiation during meiotic prophase through its participation in retinoic acid/retinoic acid receptor alpha signaling and subsequent modulation of γH2AX. Taken together, these results strongly suggest that the genetic polymorphisms of HUWE1 are closely related to spermatogenesis and non-obstructive azoospermia pathogenesis.

Characteristics of Klebsiella pneumoniae pyogenic liver abscess from 2010-2021 in a tertiary teaching hospital of South China.

OBJECTIVES: Pyogenic liver abscess (PLA) is a severe and potentially fatal infectious disease. Klebsiella pneumoniae (K. pneumoniae) is the predominant pathogen responsible for PLA. This study aims to investigate the clinical characteristics and prognostic factors of K. pneumoniae-induced pyogenic liver abscess (KP-PLA), particularly those caused by carbapenem-resistant K. pneumoniae (CRKP). METHODS: Analyses were performed on PLA patients from January 2010 to December 2021, to investigate the differences of K. pneumoniae from other etiologically infected PLA patients. Univariate and multivariate logistic regression analyses were used to compare prognostic factors between patients with carbapenem-resistant K. pneumoniae PLA (CRKP-PLA) and patients with carbapenem-sensitive K. pneumoniae PLA. RESULTS: Univariate analysis demonstrated a significant association between KP-PLA and factors including diabetes mellitus (P < 0.001), cholecystitis and cholelithiasis (P = 0.032), single abscess (P = 0.016), and abscesses with a diameter over 50 mm (P = 0.004). The CRKP group exhibited a higher prevalence of therapeutic interventions before K. pneumoniae infection, including abdominal surgery, mechanical ventilation, sputum suction, tracheal cannula, routine drainage of the abdominal cavity, and peripherally inserted central venous catheters (P < 0.05). Multivariate logistic regression analysis revealed that admission to the intensive care unit was an independent risk factor associated with CRKP-PLA (odds ratio 36; 95% confidence interval 1.77-731.56; P = 0.020). CONCLUSION: The KP-PLA patients were significantly associated with diabetes and were more likely to have single abscesses larger than 50 mm. PLA patients with a history of admission to intensive care unit or invasive therapeutic procedures should be given special consideration if combined with CRKP infection.

Tenofovir amibufenamide vs tenofovir alafenamide for treating chronic hepatitis B: A real-world study.

BACKGROUND: The efficacy and safety profile of tenofovir amibufenamide (TMF) in chronic hepatitis B (CHB) patients is not well-established. AIM: To compare the efficacy and safety of TMF and tenofovir alafenamide (TAF) over a 48-wk period in patients with CHB. METHODS: A total of 215 subjects meeting the inclusion criteria were enrolled and divided into two groups: TMF group (n = 106) and the TAF group (n = 109). The study included a comparison of virological response (VR): Undetectable hepatitis B virus DNA levels, alanine transaminase (ALT) normalization rates, renal function parameters, and blood lipid profiles. RESULTS: At 24 and 48 wk, VR rates for the TMF group were 53.57% and 78.57%, respectively, compared with 48.31% and 78.65% for the TAF group (P > 0.05). The VR rates were also similar in both groups among patients with low-level viremia, both hepatitis B e antigen (HBeAg)-positive and HBeAg-negative subgroups. The TMF cohort showed ALT normalization and renal safety profiles similar to the TAF group. There was a notable increase in total cholesterol levels in the TAF group (P = 0.045), which was not observed in the TMF group (P > 0.05). In patients with liver cirrhosis, both groups exhibited comparable VR and ALT normalization rates and renal safety profiles. However, the fibrosis 4 score at 48 wk showed a significant reduction in the TAF group as compared to the TMF group within the liver cirrhosis subgroup. CONCLUSION: Our study found TMF is as effective as TAF in treating CHB and has a comparable safety profile. However, TAF may be associated with worsening lipid profiles.