An archaeal virus-encoded anti-CRISPR protein inhibits type III-B immunity by inhibiting Cas RNP complex turnover.

CRISPR-Cas systems are widespread in prokaryotes and provide adaptive immune against viral infection. Viruses encode a type of proteins called anti-CRISPR to evade the immunity. Here, we identify an archaeal virus-encoded anti-CRISPR protein, AcrIIIB2, that inhibits Type III-B immunity. We find that AcrIIIB2 inhibits Type III-B CRISPR-Cas immunity in vivo regardless of viral early or middle-/late-expressed genes to be targeted. We also demonstrate that AcrIIIB2 interacts with Cmr4α subunit, forming a complex with target RNA and Cmr-α ribonucleoprotein complex (RNP). Furtherly, we discover that AcrIIIB2 inhibits the RNase activity, ssDNase activity and cOA synthesis activity of Cmr-α RNP in vitro under a higher target RNA-to-Cmr-α RNP ratio and has no effect on Cmr-α activities at the target RNA-to-Cmr-α RNP ratio of 1. Our results suggest that once the target RNA is cleaved by Cmr-α RNP, AcrIIIB2 probably inhibits the disassociation of cleaved target RNA, therefore blocking the access of other target RNA substrates. Together, our findings highlight the multiple functions of a novel anti-CRISPR protein on inhibition of the most complicated CRISPR-Cas system targeting the genes involved in the whole life cycle of viruses.

Unveiling the therapeutic potential of IHMT-337 in glioma treatment: targeting the EZH2-SLC12A5 axis.

Glioma is the most common malignant tumor of the central nervous system, with EZH2 playing a crucial regulatory role. This study further explores the abnormal expression of EZH2 and its mechanisms in regulating glioma progression. Additionally, it was found that IHMT-337 can potentially be a therapeutic agent for glioma. The prognosis, expression, and localization of EZH2 were determined using bioinformatics, IHC staining, Western blot (WB) analysis, and immunofluorescence (IF) localization. The effects of EZH2 on cell function were assessed using CCK-8 assays, Transwell assays, and wound healing assays. Public databases and RT-qPCR were utilized to identify downstream targets. The mechanisms regulating these downstream targets were elucidated using MS-PCR and WB analysis. The efficacy of IHMT-337 was demonstrated through IC50 measurements, WB analysis, and RT-qPCR. The effects of IHMT-337 on glioma cells in vitro were evaluated using Transwell assays, EdU incorporation assays, and flow cytometry. The potential of IHMT-337 as a treatment for glioma was assessed using a blood-brain barrier (BBB) model and an orthotopic glioma model. Our research confirms significantly elevated EZH2 expression in gliomas, correlating with patient prognosis. EZH2 facilitates glioma proliferation, migration, and invasion alongside promoting SLC12A5 DNA methylation. By regulating SLC12A5 expression, EZH2 activates the WNK1-OSR1-NKCC1 pathway, enhancing its interaction with ERM to promote glioma migration. IHMT-337 targets EZH2 in vitro to inhibit WNK1 activation, thereby suppressing glioma cell migration. Additionally, it inhibits cell proliferation and arrests the cell cycle. IHMT-337 has the potential to cross the BBB and has successfully inhibited glioma progression in vivo. This study expands our understanding of the EZH2-SLC12A5 axis in gliomas, laying a new foundation for the clinical translation of IHMT-337 and offering new insights for precision glioma therapy.

Platelet-rich plasma-derived exosomes enhance mesenchymal stem cell paracrine function and nerve regeneration potential.

BACKGROUND: Peripheral nerve injury (PNI) presents a significant clinical challenge, leading to enduring sensory-motor impairments. While mesenchymal stem cell (MSC)-based therapy holds promise for PNI treatment, enhancing its neurotrophic effects remains crucial. Platelet-rich plasma-derived exosomes (PRP-Exo), rich in bioactive molecules for intercellular communication, offer potential for modulating cellular biological activity. METHODS: PRP-Exo was isolated, and its impact on MSC viability was evaluated. The effects of PRP-Exo-treated MSCs (MSCPExo) on Schwann cells (SCs) from injured sciatic nerves and human umbilical vein endothelial cells (HUVECs) were assessed. Furthermore, the conditioned medium from MSCPExo (MSCPExo-CM) was analyzed using a cytokine array and validated through ELISA and Western blot. RESULTS: PRP-Exo enhanced MSC viability. Coculturing MSCPExo with SCs ameliorated apoptosis and promoted SC proliferation following PNI. Similarly, MSCPExo-CM exhibited pro-proliferative, migratory, and angiogenic effects. Cytokine array analysis identified 440 proteins in the MSCPExo secretome, with 155 showing upregulation and 6 showing downregulation, many demonstrating potent pro-regenerative properties. ELISA confirmed the enrichment of several angiotrophic and neurotrophic factors. Additionally, Western blot analysis revealed the activation of the PI3K/Akt signaling pathway in MSCPExo. CONCLUSION: Preconditioning MSCs with PRP-Exo enhanced the paracrine function, particularly augmenting neurotrophic and pro-angiogenic secretions, demonstrating an improved potential for neural repair.

A novel score for predicting falls in community-dwelling older people: a derivation and validation study.

BACKGROUND: Early detection of patients at risk of falling is crucial. This study was designed to develop and internally validate a novel risk score to classify patients at risk of falls. METHODS: A total of 334 older people from a fall clinic in a medical center were selected. Least absolute shrinkage and selection operator (LASSO) regression was used to minimize the potential concatenation of variables measured from the same patient and the overfitting of variables. A logistic regression model for 1-year fall prediction was developed for the entire dataset using newly identified relevant variables. Model performance was evaluated using the bootstrap method, which included measures of overall predictive performance, discrimination, and calibration. To streamline the assessment process, a scoring system for predicting 1-year fall risk was created. RESULTS: We developed a new model for predicting 1-year falls, which included the FRQ-Q1, FRQ-Q3, and single-leg standing time (left foot). After internal validation, the model showed good discrimination (C statistic, 0.803 [95% CI 0.749-0.857]) and overall accuracy (Brier score, 0.146). Compared to another model that used the total FRQ score instead, the new model showed better continuous net reclassification improvement (NRI) [0.468 (0.314-0.622), P < 0.01], categorical NRI [0.507 (0.291-0.724), P < 0.01; cutoff: 0.200-0.800], and integrated discrimination [0.205 (0.147-0.262), P < 0.01]. The variables in the new model were subsequently incorporated into a risk score. The discriminatory ability of the scoring system was similar (C statistic, 0.809; 95% CI, 0.756-0.861; optimism-corrected C statistic, 0.808) to that of the logistic regression model at internal bootstrap validation. CONCLUSIONS: This study resulted in the development and internal verification of a scoring system to classify 334 patients at risk for falls. The newly developed score demonstrated greater accuracy in predicting falls in elderly people than did the Timed Up and Go test and the 30-Second Chair Sit-Stand test. Additionally, the scale demonstrated superior clinical validity for identifying fall risk.

Effect of levels-of-processing on rates of forgetting.

The levels-of-processing (LOP) framework, proposing that deep processing yields superior retention, has provided an important paradigm for memory research and a practical means of improving learning. However, the available levels-of-processing literature focuses on immediate memory performance. It is assumed within the LOP framework that deep processing will lead to slower forgetting than will shallow processing. However, it is unclear whether, or how, the initial level of processing affects the forgetting slopes over longer retention intervals. The present three experiments were designed to explore whether items encoded at qualitatively different LOP are forgotten at different rates. In the first two experiments, depth of processing was manipulated within-participants at encoding under deep and shallow conditions (semantic vs. rhyme judgement in Experiment 1; semantic vs. consonant-vowel pattern decision in Experiment 2). Recognition accuracy (d prime) was measured between-participants immediately after learning and at 30-min, 2-h, and 24-h delays. The third experiment employed a between-participants design, contrasting the rates of forgetting following semantic and phonological (rhyme) processing at immediate, 30-min, 2-h, and 6-h delays. Results from the three experiments consistently demonstrated a large effect size of levels of processing on immediate performance and a medium-to-large level effect size on delayed recognition, but crucially no LOP × delay group interaction. Analysis of the retention curves revealed no significant differences between the slopes of forgetting for deep and shallow processing. These results suggest that the rates of forgetting are independent of the qualitatively distinct encoding operations manipulated by levels of processing.

COVID-19 infection in patients with haematological malignancies: A single-centre survey in the latest Omicron wave in China.

As the COVID-19 variant Omicron surge in Beijing, China, a better understanding of risk factors for adverse outcomes may improve clinical management in patients with haematological malignancies (HM) diagnosed with COVID-19. The study sample includes 412 cases, mainly represented by acute leukaemia, chronic myeloid leukaemia (CML), plasma cell disorders and lymphoma and chronic lymphocytic leukaemia. COVID-19 pneumonia was observed in 10.4% (43/412) of patients, and severe/critical illness was observed in 5.3% (22/412). Among the 86 cases with advanced malignancies, 17.6% (12/86) of patients developed severe/critical COVID-19, which was significantly higher than reported in patients with stable malignancies (9/326, 2.70%, p < 0.001). Similarly, the advanced malignancy cohort had a higher mortality rate (9/86, 10.5% vs. 0/326, 0%, p < 0.001) and a poor 30-day overall survival (OS) compared with the stable malignancy cohort (74.2% vs. 100.0%, p < 0.0001). Overall, nine patients (2.2%) died. The primary cause of death was progressive HM in four patients and a combination of both COVID-19 and HM in five patients. In the multivariable analysis, over 65 years of age, comorbidities and advanced malignancy were correlated with severe/critical COVID-19 in HM patients. This study sheds light on the poor outcomes among COVID-19 HM patients with the leading cause of advanced malignancy.

Characteristics and outcomes of COVID-19 in patients with plasma cell dyscrasias during the first Omicron wave in Beijing since December 2022: a retrospective study at a National Clinical Research Center for Hematologic Disease.

The wave of coronavirus disease 2019 (COVID-19) with Omicron variant reached its first peak in Beijing, China in December 2022. We delineated characteristics and factors associated with adverse outcome of patients with plasma cell dyscrasias (PCDs) and COVID-19 during the first month of the wave. A total of 104 patients with a median age of 65 years were included in the study, with multiple myeloma (74%, n=77) and primary Immunoglobulin light chain amyloidosis (16.3%, n=17) being the two most common disease. Overall, severe or critical COVID-19 was developed in 18 (17.3%) patients, with a total all-cause mortality rate of 4.8% (n=5). The vaccination coverage was 41% and 48.1%, before and during the upsurge of Omicron, respectively, calling for the improvement of vaccination in PCD patients. Multivariable analysis revealed that age was the only independent risk factors (OR=1.14, 95% CI: 1.06-1.26, p = 0.002) associated with the development of severe or critical disease. Among patients with severe or critical group, low levels of albumin (HR=18.29; 95% CI: 1.82-183.44, p = 0.013) and high levels of lactic dehydrogenase (LDH) (HR=0.08; 95% CI: 0.01-0.65, p = 0.018) were associated with longer time to negative conversion of COVID-19.

Metagenome-assembled genomes reveal carbohydrate degradation and element metabolism of microorganisms inhabiting Tengchong hot springs, China.

A hot spring is a distinctive aquatic environment that provides an excellent system to investigate microorganisms and their function in elemental cycling processes. Previous studies of terrestrial hot springs have been mostly focused on the microbial community, one special phylum or category, or genes involved in a particular metabolic step, while little is known about the overall functional metabolic profiles of microorganisms inhabiting the terrestrial hot springs. Here, we analyzed the microbial community structure and their functional genes based on metagenomic sequencing of six selected hot springs with different temperature and pH conditions. We sequenced a total of 11 samples from six hot springs and constructed 162 metagenome-assembled genomes (MAGs) with completeness above 70% and contamination lower than 10%. Crenarchaeota, Euryarchaeota and Aquificae were found to be the dominant phyla. Functional annotation revealed that bacteria encode versatile carbohydrate-active enzymes (CAZYmes) for the degradation of complex polysaccharides, while archaea tend to assimilate C1 compounds through carbon fixation. Under nitrogen-deficient conditions, there were correspondingly fewer genes involved in nitrogen metabolism, while abundant and diverse set of genes participating in sulfur metabolism, particularly those associated with sulfide oxidation and thiosulfate disproportionation. In summary, archaea and bacteria residing in the hot springs display distinct carbon metabolism fate, while sharing the common energy preference through sulfur metabolism. Overall, this research contributes to a better comprehension of biogeochemistry of terrestrial hot springs.

Risk factors of brain abscess in neonatal meningitis: a propensity score-matched study.

Brain abscess is a rare but life-threatening complication of meningitis. The purpose of this study was to identify clinical features and potentially relevant factors of brain abscess in neonates with meningitis. This study was a propensity score-matched case-control study of neonates with brain abscess and meningitis in a tertiary pediatric hospital between January 2010 and December 2020. A total of 16 neonates with brain abscess were matched to 64 patients with meningitis. Demography, clinical characteristics, laboratory results, and pathogens were collected. Conditional logistic regression analyses were performed to identify the independent risk factors associated with brain abscess. The most common pathogen we found in the brain abscess group was Escherichia coli. Risk factors of brain abscess were identified: multidrug-resistant bacterial infection (OR, 11.204; 95% CI, 2.315-54.234; p = 0.003), C-reactive protein (CRP) > 50 mg/L (OR, 11.652; 95% CI, 1.799-75.470; p = 0.010).  Conclusion: The risk factors of brain abscess are multidrug-resistant bacterial infection and CRP > 50 mg/L. Monitoring the level of CRP is essential. Bacteriological culture and rational use of antibiotics are necessary for the prevention of MDR bacterial infection as well as the occurrence of brain abscess. What is Known: • Morbidity and mortality of neonatal meningitis have declined, but brain abscess associated with neonatal meningitis is still life-threatening. What is New: • This study investigated relevant factors related to brain abscess. • It is important for neonatologists to perform prevention, early identification, and appropriate interventions for neonates with meningitis.

Investigation of the Role of T-cell Type 17 Transcription Factor in Early Diagnosis of Cholangitis Disease.

Context: Anti-mitochondrial antibody M2+ (AMA-M2+) primary bile cholangitis (PBC) is difficult to diagnose, and early diagnosis is the key to ensure effective treatment and the safety of patients. Objective: The study intended to investigate the role of T helper type 17 (Th17) cells and their transcription factors in the early diagnosis of AMA-M2+ PBC to provide an effective guarantee of the ability to predict the prognosis of patients in the future. Design: The research team designed a prospective controlled study. Setting: The study took place at the Affiliated Hospital of Hebei University in Baoding, Hebei, China. Participants: Participants were 30 patients with AMA-M2+ PBC at the hospital between November 2020 and August 2021 and 30 healthy controls who concurrently underwent physical examinations. Outcome Measures: The study measured liver function (LF) and secretion of Th17 and its transcription factors-forkhead box P3 (Foxp3) and RAR-related orphan receptor gamma (RORγt)-and inflammatory factors-interleukin-17 IL-17 and IL-22-in participants' peripheral blood. The study also evaluated Th17 and its transcription factors in AMA-M2+ PBC, determined the expression of phosphorylated proteins using Western blotting, and analyzed the relationship between Th17 and LF. Results: The Th17 in the intervention group's peripheral blood was significantly higher than that of the control group (P < .05), and the sensitivity and specificity of the AMA-M2+ PBC were 63.33% and 96.67%, respectively. The expression of Foxp3 and p-Foxp3 proteins for the intervention was significantly lower (P < .001), while RORγt and P-ROR γ T were significantly higher (P < .001). The levels of interleukin-17 (IL-17) and IL-22 for the intervention group were significantly higher than those for the control group. The Pearson correlation coefficient showed that alanine aminotransferase (ALT), alkaline phosphatase (ALP), and γ-glutamyl transpeptidase (GGT) were positively correlated with Th17 cells, RORγt, IL-17, and IL-22 and negatively correlated with Foxp3. Conclusions: Th17 plays an important role in the early diagnosis of AMA-M2+ PBC, and Th17 and its transcription factors are highly effective for the early diagnosis of AMA-M2+ PBC, which is expected to be a breakthrough in the future diagnosis of the disease.

Effective gossypol removal from cottonseed meal through optimized solid-state fermentation by Bacillus coagulans.

BACKGROUND: Cottonseed meal (CSM) is the main by-product of the cottonseed oil extraction process with high protein content, which is an important protein source for feed industry. However, CSM contains free gossypol (FG), a toxic substance that is detrimental to animal health and greatly limits its application. Microbial fermentation is currently considered to be one of the most effective methods to reduce FG and other anti-nutritional factors in CSM. Previously, yeast and bacteria species are used for degradation of FG in CSM, but showing less detoxification efficiency. Bacillus coagulans combines the properties of both lactic acid bacteria and Bacillus, producing both lactic acid and spores, and is considered a potential probiotic. In this study, we aimed to evaluate and optimize the effect of the solid-state fermentation process using a Bacillus coagulans to gossypol removal contained cottonseed meal. RESULTS: 36 B. coagulans strains were isolated and found to have the ability to remove free gossypol. Through the evaluation of strains and optimization of fermentation conditions including fermentation temperature, ratio of material to water, inoculation amount, fermentation time and pH, we have established a solid-state fermentation process using a Bacillus coagulans strain S17 on CSM substrate with 1:1 of the material-to-water ratio, 15% (v/w) seed inoculation, 2% expanded corn flour, 1% bran, and 0.3%-0.8% metal irons at 40 °C for 52 h. After fermentation, the FG content in CSM was reduced from 923.80 to 167.90 mg/kg with 81.83% detoxification efficiency. Meanwhile, the crude protein content in CSM increased from 47.98 to 52.82%, and importantly, the spore concentration of strain S17 reached 1.68 × 1010 CFU/g dry material. CONCLUSION: The study showed that B. coagulans have the potential strong ability to degrade free gossypol through cottonseed meal fermentation. This study presents a feasible process for improving the resource utilization rate and nutritional value of CSM via solid-state fermentation through B. coagulans S17.

Abdominal obesity phenotypes are associated with the risk of developing non-alcoholic fatty liver disease: insights from the general population.

BACKGROUND: The diversity of obesity-related metabolic characteristics generates different obesity phenotypes and corresponding metabolic diseases. This study aims to explore the correlation of different abdominal obesity phenotypes with non-alcoholic fatty liver disease (NAFLD). METHODS: The current study included 14,251 subjects, 7411 males and 6840 females. Abdominal obesity was defined as waist circumference ≥ 85 cm in males and ≥ 80 cm in females; according to the diagnostic criteria for metabolic syndrome recommended by the National Cholesterol Education Program Adult Treatment Panel III, having more than one metabolic abnormality (except waist circumference criteria) was defined as metabolically unhealthy. All subjects were divided into 4 abdominal obesity phenotypes based on the presence ( +) or absence (- ) of metabolically healthy/unhealthy (MH) and abdominal obesity (AO) at baseline: metabolically healthy + non-abdominal obesity (MH-AO-); metabolically healthy + abdominal obesity (MH-AO+); metabolically unhealthy + non-abdominal obesity (MH+AO-); metabolically unhealthy + abdominal obesity (MH+AO+). The relationship between each phenotype and NAFLD was analyzed using multivariate logistic regression. RESULTS: A total of 2507 (17.59%) subjects in this study were diagnosed with NAFLD. The prevalence rates of NAFLD in female subjects with MH-AO-, MH-AO+, MH+AO-, and MH+AO+ phenotypes were 1.73%, 24.42%, 7.60%, and 59.35%, respectively. Among male subjects with MH-AO-, MH-AO+, MH+AO-, and MH+AO+ phenotypes, the prevalence rates were 9.93%, 50.54%, 25.49%, and 73.22%, respectively. After fully adjusting for confounding factors, with the MH-AO- phenotype as the reference phenotype, male MH-AO+ and MH+AO+ phenotypes increased the risk of NAFLD by 42% and 47%, respectively (MH-AO+: OR 1.42, 95%CI 1.13,1.78; MH+AO+: OR 1.47, 95%CI 1.08,2.01); the corresponding risks of MH-AO+ and MH+AO+ in females increased by 113% and 134%, respectively (MH-AO+: OR 2.13, 95%CI 1.47,3.09; MH+AO+: OR 2.34, 95%CI 1.32,4.17); by contrast, there was no significant increase in the risk of NAFLD in the MH+AO- phenotype in both sexes. CONCLUSIONS: This first report on the relationship of abdominal obesity phenotypes with NAFLD showed that both MH-AO+ and MH+AO+ phenotypes were associated with a higher risk of NAFLD, especially in the female population. These data provided a new reference for the screening and prevention of NAFLD.

Comprehensive utilization of palm kernel cake for producing mannose and manno-oligosaccharide mixture and yeast culture.

Palm kernel cake (PKC) is an agricultural waste derived from palm kernel oil manufacturing, and its production is increasing year by year. It is very urgent to process this agricultural waste in an environmentally friendly way. Here, PKC was used to produce mannose and manno-oligosaccharides mixture (MMOM) and yeast culture (YC) through enzymolysis and solid-state fermentation (SSF). In enzymolysis, five factors were optimized separately and a response surface methodology analysis was performed. Then, enzymolysis of PKC was carried out at the optimal condition, and the extraction efficiency of mannose and manno-oligosaccharides reached 68.90% with mannose concentration achieving 60.27 g/L. After enzymolysis, the enzymatic hydrolysate was dried by spray drying, and the contents of MMOM reached 42.9%. In SSF, the enzymolysis residues were utilized with inoculating Saccharomyces cerevisiae for yielding YC. After optimization, the cells number of S. cerevisiae reached 2.08 × 109 cells/g and the crude protein content was increased to 27.31%. Therefore, a novel approach to produce feed additives, including MMOM and YC, with high value by comprehensive utilization of PKC was proposed, which has good application prospects in the breeding industry. KEY POINTS: • New idea for the comprehensive utilization of PKC is proposed. • PKC was used to produce mannose and mannan-oligosaccharides mixture (MMOM) by enzymolysis and spray drying. • The enzymolysis residues were reused via SSF for producing yeast culture (YC).

Protease or Clostridium butyricum addition to a low-protein diet improves broiler growth performance.

Low-protein (LP) feeds are used in the poultry industry to combat the increasing consumption of protein resources and reduce environmental pollution caused by excessive nitrogen excretion. Dietary supplementation of protease or Clostridium butyricum increases the growth performance of broilers; however, it is unclear whether they counteract the negative effects of LP diets. The effects of protease and C. butyricum on growth performance, intestinal morphology, anti-oxidant capacity, anti-inflammatory response, and microbial community of broilers have not been studied extensively. Here, 450 healthy 1-day-old Cobb500 broilers were allocated to five groups, according to different diets: basal diet (Control); LP diet (LP; 2% less crude protein than the control); LP diet + 200 g/t HuPro protease (LPH); LP diet + 1.0 × 109 CFU/t C. butyricum (LPC); and basal diet + 200 g/t oxytetracycline (Antibiotic). Supplementing both C. butyricum and protease improved the growth performance of broilers. The supplementation of HuPro protease under low-protein conditions could achieve a breeding effect similar to that of the positive control (Antibiotic). Supplementing C. butyricum could maintain intestinal barrier function, alleviate the inflammatory response, and increase ileal and cecal short-chain fatty acid concentrations. Both C. butyricum and protease altered the bacterial diversity in the cecum, increased Bacteroidetes abundance, and resulted in higher abundance of Rikenellaceae RC9 gut spp. and lower abundance of Alistipes spp. in broilers. This study demonstrates the positive effects of proteases and C. butyricum on broilers and serves as a reference for the selection of appropriate supplementation for broilers in the poultry industry. KEY POINTS: • Low-protein diet had a negative effect on growth performance of broilers. • Protease significantly reduced feed conversion rate. • Clostridium butyricum had positive effects on broilers.

Association of LDL:HDL ratio with prediabetes risk: a longitudinal observational study based on Chinese adults.

BACKGROUND: Low-density lipoprotein:high-density lipoprotein cholesterol ratio (LDL:HDL ratio) has a good performance in identifying diabetes mellitus (DM) and insulin resistance. However, it is not yet clear whether the LDL:HDL ratio is associated with a high-risk state of prediabetes. METHODS: This cohort study retrospectively analyzed the data of 100,309 Chinese adults with normoglycemia at baseline. The outcome event of interest was new-onset prediabetes. Using multivariate Cox regression and smoothing splines to assess the association of LDL:HDL ratio with prediabetes. RESULTS: During an average observation period of 37.4 months, 12,352 (12.31%) subjects were newly diagnosed with prediabetes. After adequate adjustment for important risk factors, the LDL:HDL ratio was positively correlated with the prediabetes risk, and the sensitivity analysis further suggested the robustness of the results. Additionally, in stratified analysis, we discovered significant interactions between LDL:HDL ratio and family history of DM, sex, body mass index and age (all P-interaction < 0.05); among them, the LDL:HDL ratio-related prediabetes risk decreased with the growth of body mass index and age, and increased significantly in women and people with a family history of DM. CONCLUSIONS: The increased LDL:HDL ratio in the Chinese population indicates an increased risk of developing prediabetes, especially in women, those with a family history of DM, younger adults, and non-obese individuals.

The value of combining the simple anthropometric obesity parameters, Body Mass Index (BMI) and a Body Shape Index (ABSI), to assess the risk of non-alcoholic fatty liver disease.

BACKGROUND: Body mass index (BMI) and A Body Shape Index (ABSI) are current independent risk factors for non-alcoholic fatty liver disease (NAFLD). The aim of this study was to explore the value of combining these two most common obesity indexes in identifying NAFLD. METHODS: The subjects in this study were 14,251 individuals from the NAfld in the Gifu Area, Longitudinal Analysis (NAGALA) cohort who underwent routine health examination. We integrated BMI with WC and with ABSI to construct 6 combined obesity indicators-obesity phenotypes, the combined anthropometric risk index (ARI) for BMI and ABSI, optimal proportional combination OBMI+WC and OBMI+ABSI, and multiplicative combination BMI*WC and BMI*ABSI. Several multivariable logistic regression models were established to evaluate the relationship between BMI, WC, ABSI, and the above six combined indicators and NAFLD; receiver operating characteristic (ROC) curves were drawn to compare the ability of each obesity indicator to identify NAFLD. RESULTS: A total of 2,507 (17.59%) subjects were diagnosed with NAFLD. BMI, WC, ABSI, and all other combined obesity indicators were significantly and positively associated with NAFLD in the current study, with BMI*WC having the strongest correlation with NAFLD in female subjects (OR per SD increase: 3.13) and BMI*ABSI having the strongest correlation in male subjects (OR per SD increase: 2.97). ROC analysis showed that ARI and OBMI+ABSI had the best diagnostic performance in both sexes, followed by BMI*WC (area under the curve: female 0.8912; male 0.8270). After further age stratification, it was found that ARI and multiplicative indicators (BMI*WC, BMI*ABSI) and optimal proportional combination indicators (OBMI+WC, OBMI+ABSI) significantly improved the NAFLD risk identification ability of the basic anthropometric parameters in middle-aged females and young and middle-aged males. CONCLUSION: In the general population, BMI combined with ABSI best identified obesity-related NAFLD risk and was significantly better than BMI or WC, or ABSI. We find that ARI and the multiplicative combined indicators BMI*WC and BMI*ABSI further improved risk prediction and may be proposed for possible use in clinical practice.

Prevalence of sarcopenia under different diagnostic criteria and the changes in muscle mass, muscle strength, and physical function with age in Chinese old adults.

BACKGROUND: At present, there are several diagnostic criteria of sarcopenia were used in China, and the diagnostic criteria were not unified. This study aims to investigate the consistency between the latest sarcopenia diagnostic criteria Asian Working Group for Sarcopenia(AWGS 2019) and other common diagnostic criteria. The changes of muscle mass, muscle strength and physical function with age and their effects on the diagnosis of sarcopenia were also analyzed. METHODS: A total of 1009 men aged ≥60 years were enrolled from multiple communities. Skeletal muscle mass index, grip strength and 6 m gait speed were measured. The consistency of AWGS 2019 with other diagnostic criteria was analyzed and the trends of these three indicators were observed. The differences of muscle mass, muscle strength and function among different diagnostic criteria and age groups were evaluated. In addition, the change trends of these three indicators with age were observed. RESULTS: According to AWGS 2019 diagnostic criteria, the incidence of sarcopenia in male aged 60-69 years, 70-79 years and over 80 years was 1.5%, 9.6% and 33.1%, respectively. AWGS 2019 was highly consistent with other diagnostic criteria (Kappa = 0.66-0.80, P < 0.01), except the Foundation for the National Institutes of Health(FNIH) (Kappa = 0.32, P < 0.01). When AWGSA2019 diagnostic criteria are applied, the prevalence of decreased muscle strength (39.1%) and physical function (46.4%) was significantly higher than that of low muscle mass (35.9%) in the men over 80 years old. Muscle strength (P < 0.01) and function (P < 0.01) decreased at the same rate with age, both of which were more significant than muscle mass (P < 0.01). CONCLUSION: AWGS 2019 was highly consistent with other criteria. Maintaining muscle mass should be the focus of attention before age 80, while improving muscle strength and function should be focused after age 80 to prevent disability.

A CRISPR-associated factor Csa3a regulates DNA damage repair in Crenarchaeon Sulfolobus islandicus.

CRISPR-Cas system provides acquired immunity against invasive genetic elements in prokaryotes. In both bacteria and archaea, transcriptional factors play important roles in regulation of CRISPR adaptation and interference. In the model Crenarchaeon Sulfolobus islandicus, a CRISPR-associated factor Csa3a triggers CRISPR adaptation and activates CRISPR RNA transcription for the immunity. However, regulation of DNA repair systems for repairing the genomic DNA damages caused by the CRISPR self-immunity is less understood. Here, according to the transcriptome and reporter gene data, we found that deletion of the csa3a gene down-regulated the DNA damage response (DDR) genes, including the ups and ced genes. Furthermore, in vitro analyses demonstrated that Csa3a specifically bound the DDR gene promoters. Microscopic analysis showed that deletion of csa3a significantly inhibited DNA damage-induced cell aggregation. Moreover, the flow cytometry study and survival rate analysis revealed that the csa3a deletion strain was more sensitive to the DNA-damaging reagent. Importantly, CRISPR self-targeting and DNA transfer experiments revealed that Csa3a was involved in regulating Ups- and Ced-mediated repair of CRISPR-damaged host genomic DNA. These results explain the interplay between Csa3a functions in activating CRISPR adaptation and DNA repair systems, and expands our understanding of the lost link between CRISPR self-immunity and genome stability.

DNA Motifs and an Accessory CRISPR Factor Determine Cas1 Binding and Integration Activity in Sulfolobus islandicus.

CRISPR-Cas systems empower prokaryotes with adaptive immunity against invasive mobile genetic elements. At the first step of CRISPR immunity adaptation, short DNA fragments from the invaders are integrated into CRISPR arrays at the leader-proximal end. To date, the mechanism of recognition of the leader-proximal end remains largely unknown. Here, in the Sulfolobus islandicus subtype I-A system, we show that mutations destroying the proximal region reduce CRISPR adaptation in vivo. We identify that a stem-loop structure is present on the leader-proximal end, and we demonstrate that Cas1 preferentially binds the stem-loop structure in vitro. Moreover, we demonstrate that the integrase activity of Cas1 is modulated by interacting with a CRISPR-associated factor Csa3a. When translocated to the CRISPR array, the Csa3a-Cas1 complex is separated by Csa3a binding to the leader-distal motif and Cas1 binding to the leader-proximal end. Mutation at the leader-distal motif reduces CRISPR adaptation efficiency, further confirming the in vivo function of leader-distal motif. Together, our results suggest a general model for binding of Cas1 protein to a leader motif and modulation of integrase activity by an accessory factor.

Inactivation of Target RNA Cleavage of a III-B CRISPR-Cas System Induces Robust Autoimmunity in Saccharolobus islandicus.

Type III CRISPR-Cas systems show the target (tg)RNA-activated indiscriminate DNA cleavage and synthesis of oligoadenylates (cOA) and a secondary signal that activates downstream nuclease effectors to exert indiscriminate RNA/DNA cleavage, and both activities are regulated in a spatiotemporal fashion. In III-B Cmr systems, cognate tgRNAs activate the two Cmr2-based activities, which are then inactivated via tgRNA cleavage by Cmr4, but how Cmr4 nuclease regulates the Cmr immunization remains to be experimentally characterized. Here, we conducted mutagenesis of Cmr4 conserved amino acids in Saccharolobus islandicus, and this revealed that Cmr4α RNase-dead (dCmr4α) mutation yields cell dormancy/death. We also found that plasmid-borne expression of dCmr4α in the wild-type strain strongly reduced plasmid transformation efficiency, and deletion of CRISPR arrays in the host genome reversed the dCmr4α inhibition. Expression of dCmr4α also strongly inhibited plasmid transformation with Cmr2αHD and Cmr2αPalm mutants, but the inhibition was diminished in Cmr2αHD,Palm. Since dCmr4α-containing effectors lack spatiotemporal regulation, this allows an everlasting interaction between crRNA and cellular RNAs to occur. As a result, some cellular RNAs, which are not effective in mediating immunity due to the presence of spatiotemporal regulation, trigger autoimmunity of the Cmr-α system in the S. islandicus cells expressing dCmr4α. Together, these results pinpoint the crucial importance of tgRNA cleavage in autoimmunity avoidance and in the regulation of immunization of type III systems.