New Insights on the Therapeutic Potential of Runt-Related Transcription Factor 2 for Osteoarthritis: Evidence from Mendelian Randomization.

INTRODUCTION: Research has highlighted the role of runt-related transcription factor 2 (Runx2) in the development of osteoarthritis (OA); however, its causal association remains unclear. This study aimed to explore whether Runx2 expression is causally associated with OA and assess its therapeutic potential for OA. METHODS: Genetic proxy instruments for Runx2 expression were obtained from gene expression quantitative trait locus (eQTLs) study of eQTLGen Consortium (n = 31,684). Aggregated genome-wide association study (GWAS) data for OA (including all OA [177,517 cases and 649,173 controls], knee OA (KOA) [62,497 cases and 333,557 controls], and hip OA (HOA) [36,445 cases and 316,943 controls]) were extracted from the Genetics of Osteoarthritis Consortium. We integrated eQTLs data with OA GWAS data to estimate their causal association and to estimate the potential of Runx2 as a drug target in the treatment of OA using summary data-based Mendelian randomization (SMR) analysis. Furthermore, different OA GWAS data (including all OA [77,052 cases and 378,169 controls], KOA [24,955 cases and 378,169 controls], and HOA [15,704 cases and 378,169 controls]) derived from the GWAS Catalog database were used for replication study. RESULTS: SMR analysis showed that high expression levels of Runx2 were associated with an increased risk of all OA [odds ratio (OR) 1.044, 95% confidence interval (CI) 1.023-1.067; P = 5.03 × 10-5], KOA (OR 1.040, 95% CI 1.006-1.075; P = 0.021), and HOA (OR 1.067, 95% CI 1.022-1.113; P = 0.003). This suggests that Runx2 inhibitors may have promising potential for the treatment of OA. Notably, the causal effects of Runx2 with all OA (OR 1.053, 95% CI 1.027-1.079; P = 3.95 × 10-5) and KOA (OR 1.043, 95% CI 1.001-1.087; P = 0.045) were repeated in the replication study, but limited evidence supported the association of Runx2 expression levels with HOA (OR 1.045, 95% CI 0.993-1.101; P = 0.094). CONCLUSIONS: Our analyses indicate a positive correlation between Runx2 expression and OA risk across all three phenotypes, suggesting the potential of Runx2 inhibitors in the treatment of OA and providing evidence from a genetic perspective.

Skeletal interoception in osteoarthritis.

The interoception maintains proper physiological conditions and metabolic homeostasis by releasing regulatory signals after perceving changes in the internal state of the organism. Among its various forms, skeletal interoception specifically regulates the metabolic homeostasis of bones. Osteoarthritis (OA) is a complex joint disorder involving cartilage, subchondral bone, and synovium. The subchondral bone undergoes continuous remodeling to adapt to dynamic joint loads. Recent findings highlight that skeletal interoception mediated by aberrant mechanical loads contributes to pathological remodeling of the subchondral bone, resulting in subchondral bone sclerosis in OA. The skeletal interoception is also a potential mechanism for chronic synovial inflammation in OA. In this review, we offer a general overview of interoception, specifically skeletal interoception, subchondral bone microenviroment and the aberrant subchondral remedeling. We also discuss the role of skeletal interoception in abnormal subchondral bone remodeling and synovial inflammation in OA, as well as the potential prospects and challenges in exploring novel OA therapies that target skeletal interoception.

Molecular characterization, expression and antibacterial function of a macin, HdMac, from Haliotis discus hannai.

Macins are a family of antimicrobial peptides, which play multiple roles in the elimination of invading pathogens. In the present study, a macin was cloned and characterized from Pacific abalone Haliotis discus hannai (Designated as HdMac). Analysis of the conserved domain suggested that HdMac was a new member of the macin family. In non-stimulated abalones, HdMac transcripts were constitutively expressed in all five tested tissues, especially in hemocytes. After Vibrio harveyi stimulation, the expression of HdMac mRNA in hemocytes was significantly up-regulated at 12 hr (P < 0.01). RNAi-mediated knockdown of HdMac transcripts affected the survival rates of abalone against V. harveyi. Moreover, recombinant protein of HdMac (rHdMac) exhibited high antibacterial activities against invading bacteria, especially for Vibrio anguillarum. In addition, rHdMac possessed binding activities towards glucan, lipopolysaccharides (LPS), and peptidoglycan (PGN), but not chitin in vitro. Membrane integrity analysis revealed that rHdMac could increase the membrane permeability of bacteria. Meanwhile, both the phagocytosis and chemotaxis ability of hemocytes could be significantly enhanced by rHdMac. Overall, the results showed that HdMac could function as a versatile molecule involved in immune responses of H. discus hannai.

Revisiting prostaglandin E2: A promising therapeutic target for osteoarthritis.

Osteoarthritis (OA) is a complex disease characterized by cartilage degeneration and persistent pain. Prostaglandin E2 (PGE2) plays a significant role in OA inflammation and pain. Recent studies have revealed the significant role of PGE2-mediated skeletal interoception in the progression of OA, providing new insights into the pathogenesis and treatment of OA. This aspect also deserves special attention in this review. Additionally, PGE2 is directly involved in pathologic processes including aberrant subchondral bone remodeling, cartilage degeneration, and synovial inflammation. Therefore, celecoxib, a commonly used drug to alleviate inflammatory pain through inhibiting PGE2, serves not only as an analgesic for OA but also as a potential disease-modifying drug. This review provides a comprehensive overview of the discovery history, synthesis and release pathways, and common physiological roles of PGE2. We discuss the roles of PGE2 and celecoxib in OA and pain from skeletal interoception and multiple perspectives. The purpose of this review is to highlight PGE2-mediated skeletal interoception and refresh our understanding of celecoxib in the pathogenesis and treatment of OA.

Intrinsic and extrinsic pathways of apoptosis induced by multiple antibiotics residues and ocean acidification in hemocytes of scallop Argopecten irradians irradians: An interactionist perspective.

The increasing prevalence of antibiotics in seawater across global coastal areas, coupled with the ocean acidification induced by climate change, present a multifaceted challenge to marine ecosystems, particularly impacting the key physiological processes of marine organisms. Apoptosis is a critical adaptive response essential for maintaining cellular homeostasis and defending against environmental threats. In this study, bay scallops Argopecten irradians irradians were exposed to multiple antibiotics (sulfamethoxazole, tetracycline, oxytetracycline, norfloxacin, and erythromycin, each at a concentration of 1 µg/L) combined with/without acidic seawater (pH 7.6) for 35 days. The single and interactive effects of the two stressors on apoptosis and the underlying mechanisms in hemocytes of A. irradians irradians were determined through flow cytometry analysis, comet assay, oxidative stress biomarkers analysis, and transcriptome analysis. Results showed that apoptosis could be triggered by either AM exposure or OA exposure, but through different pathways. Exposure to AM leads to mitochondrial dysfunction and oxidative damage, which in turn triggers apoptosis via a series of cellular events in both intrinsic and extrinsic pathways. Conversely, while OA exposure similarly induced apoptosis, its effects are comparatively subdued and are predominantly mediated through the intrinsic pathway. Additionally, the synergistic effects of AM and OA exposure induced pronounced mitochondrial dysfunction and oxidative damages in the hemocytes of A. irradians irradians. Despite the evident cellular distress and the potential initiation of apoptotic pathways, the actual execution of apoptosis appears to be restrained, which might be attributed to an energy deficit within the hemocytes. Our findings underscore the constrained tolerance capacity of A. irradians irradians when faced with multiple environmental stressors, and shed light on the ecotoxicity of antibiotic pollution in the ocean under prospective climate change scenarios.

Interactive effects of multiple antibiotic residues and ocean acidification on physiology and metabolome of the bay scallops Argopecten irradians irradians.

Coastal areas are confronted with compounding threats arising from both climatic and non-climatic stressors. Antibiotic pollution and ocean acidification are two prevalently concurrent environmental stressors. Yet their interactive effects on marine biota have not been investigated adequately and the compound hazard remain obscure. In this study, bay scallops Argopecten irradians irradians were exposed to multiple antibiotics (sulfamethoxazole, tetracycline, oxytetracycline, norfloxacin, and erythromycin, each at a concentration of 1 µg/L) combined with/without acidic seawater (pH 7.6) for 35 days. The single and interactive effects of the two stressors on A. irradians irradians were determined from multidimensional bio-responses, including energetic physiological traits as well as the molecular underpinning (metabolome and expressions of key genes). Results showed that multiple antibiotics predominantly enhanced the process of DNA repair and replication via disturbing the purine metabolism pathway. This alternation is perhaps to cope with the DNA damage induced by oxidative stress. Ocean acidification mainly disrupted energy metabolism and ammonia metabolism of the scallops, as evidenced by the increased ammonia excretion rate, the decreased O:N ratio, and perturbations in amino acid metabolism pathways. Moreover, the antagonistic effects of multiple antibiotics and ocean acidification caused alternations in the relative abundance of neurotransmitter and gene expression of neurotransmitter receptors, which may lead to neurological disorders in scallops. Overall, the revealed alternations in physiological traits, metabolites and gene expressions provide insightful information for the health status of bivalves in a natural environmental condition under the climate change scenarios.

The Albumin to Globulin Ratio Performs Well for Diagnosing Periprosthetic Joint Infection: A Single-Center Retrospective Study.

BACKGROUND: Accurate diagnosis of the periprosthetic joint infection (PJI) remains a challenge for surgeons. The purpose of this study was to assess the value of albumin to globulin ratio (AGR) and globulin (GLB) for diagnosing PJI. METHODS: A total of 182 patients undergoing revision after arthroplasty were included and divided into 2 groups, 61 in knee group (PJI: 38; non-PJI: 23) and 121 in hip group (PJI: 26; non-PJI: 95). We used receiver operating characteristic curves to determine the diagnostic value of AGR, GLB, inflammatory markers (erythrocyte sedimentation rate [ESR] and C-reactive protein [CRP]). RESULTS: The receiver operating characteristic curves showed the areas under the curve of AGR, GLB, ESR, and CRP in the knee group were 0.940, 0.928, 0.867, and 0.848, respectively, and they were 0.855, 0.831, 0.886, and 0.912 in the hip group. The optimal predictive cut-off values for AGR in knee and hip groups were 1.375 and 1.295, respectively. The sensitivity and specificity of AGR, respectively, were 94.7% and 87.0% (knee group) and 84.6% and 75.8% (hip group) for diagnosing PJI. The sensitivity of "AGR or ESR" and specificity of "AGR and GLB" in the knee group were 99.6% and 98.9%, respectively. CONCLUSION: For knee or hip groups, the AGR exhibits good value for the diagnosis of PJI comparable with ESR and CRP. The AGR and GLB, together with CRP and ESR, should be used as the preferred indicators for diagnosing PJI. The "AGR or ESR" and "AGR and GLB" in the knee group have an excellent diagnostic value in sensitivity and specificity, respectively.

[Correlation between preoperative MRI measurement of cross-sectional area of hamstring tendon and graft in anterior cruciate ligament reconstruction].

OBJECTIVE: To study the corretation between the cross-sectional area of hamstring tenden measured by MRI and gragt in anterior cruciate ligament rexonstruction. METHODS: MRI data of 50 patients who planned to undergo anterior cruciate ligament reconstruction from November 2021 to March 2022 were collected, including 32 males and 18 females, aged from 19 to 48 years old with an average of(31.1±8.7) years. Before the operation, the semitendinosus and gracilis tendons were measured and recorded by MRI, and then the anterior cruciate ligament was reconstructed under arthroscope. During the operation, gracilis and semitendinosus tendons were taken to prepare the final tendon to be transplanted, and the diameter of the prepared final graft was measured during the operation. Finally, the data were analyzed by statistical software. RESULTS: The cross sectional areas of semitendinosus tendon, gracilis tendon, semitendinosus tendon and gracilis tendon measured by MRI were significantly and positively correlated with the diameter of grafts required in anterior cruciate ligament surgery, the r values were 0.858, 0.728, 0.842(P<0.001), respectively. The area under curre (AUC), sensitivity, and specificity of the sum of the cross sectional areas of semitendinosus tendon and gracilis tendon were 0.925, 90.48%, and 85.71%, respectively. CONCLUSION: In patients undergoing anterior cruciate ligament reconstruction, preoperative MRI measurement has a strong statistical correlation with the diameter of hamstring muscle transplantation during operation. The sum of the cross sectional areas of semitendinosus tendon and gracilis tendon has a high predictive value for the diameter of grafts during anterior cruciate ligament reconstruction, and can predict the size of grafts during operation.

Effects of exposure to nanoplastics on the gill of mussels Mytilus galloprovincialis: An integrated perspective from multiple biomarkers.

The pervasive presence of nanoplastics (NPs) in marine environments poses a threat to marine organisms. Gills, as the organ in direct contact with the environment in marine invertebrates, maybe the first to accumulate NPs. To date, the toxic effects of NPs on the gills of marine invertebrates are still largely unknown. In this study, the response of multiple biomarkers (i.e., total antioxidant capacity, the activity of acetylcholine, ion content and transport enzyme, metabolic enzymes, and lipids content) in mussels Mytilus galloprovincialis exposed to polystyrene nanoplastics (PS-NPs) for 7 days were evaluated. Significant inductions of total antioxidant capacity (T-AOC) and inhibition of acetylcholine (AChE) activity were detected after 7 days of PS-NPs exposure. PS-NPs also triggered significant alteration in ion content (Na+ and K+) and suppressed the activities of the ion transport enzyme (Na+/K+-ATPase). Moreover, we found the activity of metabolic enzymes (succinate dehydrogenase and pyruvate kinase) and lipids content (triacylglycerol and cholesterol) were significantly altered, suggesting the interference of PS-NPs on energy metabolism and lipid metabolism. This investigation provides substantial information to understand the physical responses of invertebrate gills to PS-NPs exposure. Given the crucial ecological roles of invertebrates, the presence of PS-NPs in the marine environment may have far-reaching impacts on population abundance, biodiversity, and stability of the marine ecosystem.

Comparison of spatiotemporal, kinematic, and kinetic gait characteristics in total and unicompartmental knee arthroplasty during level walking: A systematic review and meta-analysis.

PURPOSE: This meta-analysis was performed to compare the spatiotemporal, kinematic, and kinetic gait characteristics during level walking between total knee arthroplasty (TKA) and unicompartmental knee arthroplasty (UKA). METHODS: An electronic database literature search was performed to screen clinical trials which were included the studies evaluating not only spatiotemporal, kinematic, and kinetic gait parameters, but also knee range of motion and knee score (Knee Society Score and Oxford Knee Score, i.e., KSS and OKS). The data analysis was performed using statistical software Stata 14.0 and Review Manager 5.4. RESULTS: Thirteen studies (369 knees) that met the criteria were eventually included in this meta-analysis. The results revealed significant differences between UKA and TKA with regard to walking speed (P = 0.04), stride length (P = 0.02), maximum knee flexion at loading (P = 0.001), the 1st peak of vert-GRF (P = 0.006), the 1st valley of vert-GRF (P = 0.007), knee internal rotational moment (P = 0.04), knee extension (P < 0.00001), and KSS Function score (P = 0.05). In contrast, there were no statistical differences in the remaining spatiotemporal, kinematic, and kinetic gait parameters. CONCLUSION: Medial UKA design is superior to TKA design with regard to walking speed, stride length, maximum knee flexion at loading, the 1st peak and the 1st valley of vert-GRF, knee internal rotational moment, knee extension, and KSS Function score. And it could provide a stronger basis for physicians to make clinical decisions.

In vitro study of deltamethrin-induced extracellular traps in hemocytes of Ruditapes philippinarum.

Deltamethrin (DLM), a broad-spectrum pesticide, has been proven to have toxic effects on aquatic organisms. Here, we detected the formation of extracellular traps (ETosis) formation in Manila clam (Ruditapes philippinarum) hemocytes stimulated by three concentrations of DLM (0.01, 0.1 and 1 µg/mL) in vitro, and explored the underlying mechanisms induced by this pesticide. Extracellular DNA structure observation and quantitative results indicated that DLM exposure could obviously induce hemocytes ETosis, especially under high concentration of DLM induction. Moreover, DLM increased the levels of myeloperoxidase (MPO) and reactive oxygen species (ROS) in a dose-dependent manner, and enhanced the mRNA expression of several ROS-related genes. DPI (NADPH oxidase inhibitor) and ABAH (MPO inhibitor) could substantially inhibit DLM-induced extracellular traps (ETs), suggesting that the induced ETs release was caused by the induction of the ROS burst and MPO production. In addition, three concentrations of DLM-induced ETs were also accompanied by mitochondrial dysfunction, such as increasing the production of mitochondrial ROS, leading to a decrease in mitochondrial membrane potential (MMP) and activation of mitochondrial permeability transition pore (MPTP). Taken together, these results will shed new light on the immunotoxicity of DLM in clams and perhaps lays the foundation for health assessment in bivalves.

Intestinal microbiota perturbations in the gastropod Trochus niloticus concurrently exposed to ocean acidification and environmentally relevant concentrations of sulfamethoxazole.

Ocean acidification (OA) and antibiotic pollution pose severe threats to the fitness of keystone species in marine ecosystems. However, the combined effects of OA and antibiotic pollution on the intestinal microbiota of marine organisms are still not well known. In this study, we exposed the herbivorous gastropod Trochus niloticus, a keystone species to maintains the stability of coral reef ecosystems, to acidic seawater (pH 7.6) and/or sulfamethoxazole (SMX, 100 ng/L, 1000 ng/L) for 28 days and determined their impacts on (1) the accumulation of SMX in the intestine of T. niloticus; (2) the characteristics of the intestinal microbiota in T. niloticus; (3) the relative abundances of sulfonamide resistance genes (i.e., sul1 and sul2) and intI1 in the intestinal microbiota of T. niloticus. Our results show that OA exposure leads to dramatic microbiota dysbiosis in the intestine of T. niloticus, including changes in bacterial community diversity and structure, decreased abundances of dominant species, existences of characteristic taxa, and altered functional predictions. In addition, SMX exposure at environmentally relevant concentrations had little effect on the intestinal microbiota of T. niloticus, whether in isolation or in combination with OA. However, after exposure to the higher SMX concentration (1000 ng/L), the accumulation of SMX in the intestine of T. niloticus could induce an increase in the copies of sul2 in the intestinal microbiota. These results suggest that the intestinal health of T. niloticus might be affected by OA and SMX, which might lead to fitness loss of the keystone species in coral reef ecosystems.

Characteristic and antibacterial effect of a histone H2A and its preliminary roles in extracellular traps in manila clam Ruditapes philippinarum.

In the present study, a histone H2A (designed as RpH2A) was identified and characterized from clam Ruditapes philippinarum, and its open reading frame (ORF) was of 387 bp encoding a polypeptide of 128 amino acids. The deduced amino acid sequence of RpH2A shared high identities ranging from 57.1% to 96.1% with that of other identified H2A. The mRNA expression of RpH2A was up-regulated significantly after Vibrio anguillarum challenge. The recombinant RpH2A protein (rRpH2A) displayed significantly binding affinity to lipopolysaccharide (LPS) and peptidoglycan (PGN) in vitro, and also exhibited antimicrobial properties against Escherichia coli. In addition, the antimicrobial RpH2A was shown to co-localize with extracellular traps (ETs) released from hemocytes induced by E. coli, suggesting that RpH2A might contribute to eliminate invading bacteria in clam ETs. Altogether, our data revealed that RpH2A could function as antimicrobial peptides, which might play a crucial role in the immune responses of hemocytes ETs in clams.

Differential expression of HAVCR2 gene in pan-cancer: A potential biomarker for survival and immunotherapy.

T-cell immunoglobulin mucin 3 (TIM-3) has emerged as a promising immune checkpoint target in cancer therapy. However, the profile of the hepatitis A virus cellular receptor 2 (HAVCR2) gene, encoding TIM-3 expression, is still obscure, along with its role in cancer immunity and prognosis. This study comprehensively analyzed HAVCR2 expression patterns in pan-cancer and underlined its potential value for immune checkpoint inhibitor-based immunotherapy. Our results displayed that HAVCR2 was differentially expressed and closely corresponded to survival status in pan-cancer. More importantly, the HAVCR2 expression level was also significantly related to cancer immune infiltration, immune checkpoint genes, and immune marker genes. Enrichment analyses implicated HAVCR2-associated terms in cancer, including immunity, metabolism, and inflammation. Our study demonstrated that HAVCR2 could participate in differing degrees of immune infiltration in tumorigenesis. The highlights of the HAVCR2 pathway revealed that TIM-3 could function as both a biomarker and clinical target to improve the therapeutic efficacy of immunotherapy.

A Pan-Cancer Analysis of the Oncogenic Role of BCL7B: A Potential Biomarker for Prognosis and Immunotherapy.

Background: Previous studies have partly explored the role of B-cell CLL/lymphoma 7 protein family member B (BCL7B) in tumorigenesis and development. However, the prognosis and immunoregulatory value of BCL7B in pan-cancer patients remains unclear. Methods: Through The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases, the distinct expression of BCL7B gene in 33 tumors and adjacent normal tissues was analyzed. The Kaplan-Meier method (univariate Cox regression analysis and Kaplan-Meier curve) was used to identify the cancer types whose BCL7B gene expression was related to prognosis. The receiver operating characteristic (ROC) curve was used to elucidate the diagnosis value of BCL7B gene. Spearman's rank correlation coefficient was used to explore the relationship between BCL7B gene expression and immune cell infiltration, immune checkpoints, DNA methylation, DNA repair genes, immune-activating genes, immune-suppressing genes, immune subtypes, tumor mutation burden (TMB), and microsatellite instability (MSI). The Wilcoxon rank sum test and Kruskal-Wallis test were used to compare the expression of BCL7B gene in tumor tissues with different clinicopathological features. Gene set enrichment analysis (GSEA) was conducted to identify the tumor-related pathways in pan-cancer. The Human Protein Atlas (HPA) database was used to verify the BCL7B gene expression at the protein level. Results: High expression of BCL7B was associated with an inferior prognosis in glioblastoma multiforme (GBM), glioma (GBMLGG), kidney chromophobe (KICH), brain lower grade glioma (LGG), oral squamous cell carcinoma (OSCC), rectum adenocarcinoma (READ), and uveal melanoma (UVM). Low expression of BCL7B was associated with a poor prognosis in kidney renal clear cell carcinoma (KIRC), kidney renal papillary cell carcinoma (KIRP), skin cutaneous melanoma (SKCM), thyroid carcinoma (THCA), and sarcoma (SARC). The BCL7B gene expression had varying degrees of correlation with 24 immune cell subsets in 37 tumor environments such as adrenocortical carcinoma (ACC) and bladder urothelial carcinoma (BCLA). Spearman's rank correlation coefficient showed that BCL7B gene expression had different degrees of correlation with 47 immune checkpoints, 46 immune-activating genes, 24 immune-suppressing genes, 5 DNA repair genes, and DNA methylation, TMB, and MSI in 39 tumors. GSEA suggested that BCL7B was notably associated with cancer-related and immune-related pathways. Conclusion: In summary, BCL7B gene has a high diagnostic and prognostic value in pan-cancer and is related to the infiltration of 24 immune cell subsets in pan-cancer.

Immunomodulatory mechanisms of abatacept: A therapeutic strategy for COVID-19.

Coronavirus disease 2019 (COVID-19) caused by coronavirus-2 (SARS-CoV-2) infection has rapidly spread throughout the world and become a major threat to human beings. Cytokine storm is a major cause of death in severe patients. Abatacept can suppress cytokines used as antirheumatic drugs in clinical applications. This study analyzed the molecular mechanisms of abatacept treatment for COVID-19. Differentially expressed genes (DEGs) were identified by analyzing expression profiling of abatacept treatment for rheumatoid arthritis (RA) patients and SARS-CoV-2 infection patients. We found that 59 DEGs were upregulated in COVID-19 patients and downregulated following abatacept treatment. Gene set enrichment analysis (GSEA) and Gene Ontology (GO) analysis showed that immune and inflammatory responses were potential regulatory mechanisms. Moreover, we verified 8 targeting genes and identified 15 potential drug candidates for the treatment of COVID-19. Our study illustrated that abatacept could be a promising property for preventing severe COVID-19, and we predicted alternative potential drugs for the treatment of SARS-CoV-2 infection.

A single-CRD C-type lectin from Haliotis discus hannai acts as pattern recognition receptor enhancing hemocytes opsonization.

C-type lectins (CTLs), as a member of the Ca2+-dependent carbohydrate recognition protein superfamily, play multiple roles in non-self recognition and the elimination of invading pathogens. In this study, a C-type lectin was identified and characterized from the Pacific abalone Haliotis discus hannai (designed as HdClec), and its open reading frame (ORF) encoded a polypeptide of 163 amino acids containing a typical signal peptide and only one carbohydrate-recognition domain (CRD). The deduced amino acid sequence of CRD in HdClec shared identities ranging from 22.4% to 39.8% with that of other identified CRDs of CTLs. A novel NPN motif was found in Ca2+-binding site 2 of HdClec. The mRNA transcripts of HdClec were detectable in all the examined tissues of non-stimulated abalones, with the highest expression in hepatopancreas (224.13-fold of that in gills). The expression of HdClec mRNA in hemocytes was significantly up-regulated after Vibrio harveyi challenge. Recombinant HdClec protein (rHdClec) could bind lipopolysaccharide (LPS) and peptidoglycan (PGN) in vitro in the presence of Ca2+. Coinciding with the PAMPs binding assay, rHdClec displayed broad agglutination activities towards Gram-negative bacteria V. splendidus, V. anguillarum, V. parahaemolyticus, V. harveyi, Escherichia coli, and Gram-positive bacteria Micrococcus luteus. Moreover, rHdClec could significantly elicit the chemotactic response of hemocytes in vitro. And the phagocytosis and encapsulation ability of hemocytes could be significantly enhanced by rHdClec. All these results showed that HdClec could function as pattern recognition receptors (PRRs) and further enhance the opsonization of hemocytes, which might play a crucial role in the innate immune responses of Pacific abalone.

Effects of the potential probiotic Bacillus subtilis D1-2 on growth, digestion, immunity and intestinal flora in juvenile sea cucumber, Apostichopus japonicus.

In the present study, a potential probiotic Bacillus subtilis D1-2 with antibacterial activity was isolated from the gut of Apostichopus japonicus. The purpose of this experiment was to assess the effect of B. subtilis D1-2 at different concentrations (C: 0 CFU/g, BL: 105 CFU/g, BM: 107 CFU/g and BH: 109 CFU/g) on the growth performance, digestive enzyme activity, immune ability and intestinal flora of A. japonicus. After the 56-day feeding trial, the final body weight and weight gain rate of juvenile sea cucumber A. japonicus fed B. subtilis D1-2 were significantly increased, especially in the BM group. Additionally, the lipase activity of the intestine was significantly increased in the BM and BH groups. Enhanced immunity was also found in sea cucumbers supplemented with B. subtilis D1-2. Alpha diversity indices showed that the B. subtilis D1-2-supplemented groups had higher intestinal microbial richness and diversity than the control group. The beta diversity analysis indicated that the bacterial communities in the B. subtilis D1-2-supplemented groups were quite similar but different from the bacterial communities in the control group. Dietary supplementation with B. subtilis D1-2 increased the relative abundance of some potential probiotic-related genera, including Lactobacillus, Clostridium, Lactococcus, Bifidobacterium and Streptococcus. In conclusion, dietary addition of B. subtilis D1-2 could effectively promote the growth of A. japonicus, improve its digestion and immunity capacity to a certain extent, and actively regulate the intestinal microflora of A. japonicus.

Comparison of posterior cruciate retention and substitution in total knee arthroplasty during gait: a systematic review and meta-analysis.

BACKGROUND: To compare the gait patterns between posterior cruciate retention and substitution in total knee arthroplasty (TKA). METHODS: Electronic databases including the PubMed, Embase, CINAHL, Web of Science, and Cochrane databases were searched to identify clinical trials investigating posterior cruciate retention versus substitution in TKA. The outcome measurements were the kinematic gait parameters (flexion at heel strike, maximum flexion during loading response, flexion range during loading, minimal flexion at terminal stance, maximal flexion at the swing, and total flexion during the gait cycle), Knee Society Score (KSS), knee flexion, knee extension, and walking speed. Statistical software Review Manager 5.4 and Stata 14.0 were used for data analysis. RESULTS: There were finally 9 studies included in this meta-analysis. The results did not reveal differences between posterior cruciate retention (CR) and posterior cruciate substitution (PS) groups in TKA, in terms of kinematic gait parameters, knee extension, walking speed, and KSS. However, the PS group had a significantly larger knee flexion angle than that in the CR group [weighted mean difference = - 3.20, 95% CI - 6.13 to - 0.28, P = 0.03]. CONCLUSION: Both the posterior cruciate retention and posterior cruciate substitution lead to obvious improvements in patient function and have their advantages in getting a good cup position. The PS design is significantly better on the knee flexion, while there are no statistical differences in kinematic gait parameters and outcome scores between them. This might indicate that surgeons do not necessarily need a PS design to substitute the posterior cruciate ligament during TKA.

Identification, antibacterial activities and action mode of two macins from manila clam Venerupis philippinarum.

In this study, two macins were identified from clam Venerupis philippinarum (designated as VpMacin-1 and VpMacin-2). They showed 64.71% similarity with each other. The highest mRNA expression of VpMacin-1 and VpMacin-2 was detected in gills and hepatopancreas, respectively, in non-stimulated clams, and their expression could be induced significantly in hemocytes after Vibrio anguillarum infection. Silencing of VpMacin-1 and VpMacin-2 led to 22% and 49% mortality 6 days post infection. Escherichia coli cells were killed by recombinant protein rVpMacin-1 and rVpMacin-2 within 1000 and 400 min, respectively, at a concentration of 1.0 × MIC. Compared with rVpMacin-1, rVpMacin-2 not only showed higher broad-spectrum antimicrobial activities towards Vibrio strains, but possessed stronger abilities to inhibit the formation of bacterial biofilm. Both membrane integrity and electrochemical assay indicated that rVpMacins were capable of causing bacterial membrane permeabilization, especially for rVpMacin-2. Besides, rVpMacin-1 significantly induced both phagocytic (0.1 and 1.0 × MIC, p < 0.05) and chemotactic effects (0.1 × MIC, p < 0.01) of hemocytes, while there was no significant increase for rVpMacin-2. Overall, our results suggested that VpMacin-1 and VpMacin-2 play important roles in host defense against invasive pathogens.