Commensal bacteria direct selective cargo sorting to promote symbiosis.

Mucosal immunity protects a host from intestinal inflammation and infection and is profoundly influenced by symbiotic bacteria. Here we report that in mice symbiotic bacteria directed selective cargo sorting in Paneth cells to promote symbiosis through Nod2, a cytosolic bacterial sensor, and the multifunctional protein kinase LRRK2, both encoded by inflammatory bowel disease (IBD)-associated genes. Commensals recruited Nod2 onto lysozyme-containing dense core vesicles (DCVs), which was required for DCV localization of LRRK2 and a small GTPase, Rab2a. Deficiency of Nod2, LRRK2 or Rab2a or depletion of commensals resulted in lysosomal degradation of lysozyme. Thus, commensal bacteria and host factors orchestrate the lysozyme-sorting process to protect the host from enteric infection, implicating Paneth cell dysfunction in IBD pathogenesis.

Construction of LncRNA-Related ceRNA Networks in Longissimus Dorsi Muscle of Jinfen White Pigs at Different Developmental Stages.

The development of skeletal muscle in pigs might determine the quality of pork. In recent years, long non-coding RNAs (lncRNAs) have been found to play an important role in skeletal muscle growth and development. In this study, we investigated the whole transcriptome of the longissimus dorsi muscle (LDM) of Jinfen White pigs at three developmental stages (1, 90, and 180 days) and performed a comprehensive analysis of lncRNAs, mRNAs, and micro-RNAs (miRNAs), aiming to find the key regulators and interaction networks in Jinfen White pigs. A total of 2638 differentially expressed mRNAs (DE mRNAs) and 982 differentially expressed lncRNAs (DE lncRNAs) were identified. Compared with JFW_1d, there were 497 up-regulated and 698 down-regulated DE mRNAs and 212 up-regulated and 286 down-regulated DE lncRNAs in JFW_90d, respectively. In JFW_180d, there were 613 up-regulated and 895 down-regulated DE mRNAs and 184 up-regulated and 131 down-regulated DE lncRNAs compared with JFW_1d. There were 615 up-regulated and 477 down-regulated DE mRNAs and 254 up-regulated and 355 down-regulated DE lncRNAs in JFW_180d compared with JFW_90d. Compared with mRNA, lncRNA has fewer exons, fewer ORFs, and a shorter length. We performed GO and KEGG pathway functional enrichment analysis for DE mRNAs and the potential target genes of DE lncRNAs. As a result, several pathways are involved in muscle growth and development, such as the PI3K-Akt, MAPK, hedgehog, and hippo signaling pathways. These are among the pathways through which mRNA and lncRNAs function. As part of this study, bioinformatic screening was used to identify miRNAs and DE lncRNAs that could act as ceRNAs. Finally, we constructed an lncRNA-miRNA-mRNA regulation network containing 26 mRNAs, 7 miRNAs, and 17 lncRNAs; qRT-PCR was used to verify the key genes in these networks. Among these, XLOC_022984/miR-127/ENAH and XLOC_016847/miR-486/NRF1 may function as key ceRNA networks. In this study, we obtained transcriptomic profiles from the LDM of Jinfen White pigs at three developmental stages and screened out lncRNA-miRNA-mRNA regulatory networks that may provide crucial information for the further exploration of the molecular mechanisms during skeletal muscle development.

The gut microbiome modulates the transformation of microglial subtypes.

Clinical and animal studies have shown that gut microbiome disturbances can affect neural function and behaviors via the microbiota-gut-brain axis, and may be implicated in the pathogenesis of several brain diseases. However, exactly how the gut microbiome modulates nervous system activity remains obscure. Here, using a single-cell nucleus sequencing approach, we sought to characterize the cell type-specific transcriptomic changes in the prefrontal cortex and hippocampus derived from germ-free (GF), specific pathogen free, and colonized-GF mice. We found that the absence of gut microbiota resulted in cell-specific transcriptomic changes. Furthermore, microglia transcriptomes were preferentially influenced, which could be effectively reversed by microbial colonization. Significantly, the gut microbiome modulated the mutual transformation of microglial subpopulations in the two regions. Cross-species analysis showed that the transcriptome changes of these microglial subpopulations were mainly associated with Alzheimer's disease (AD) and major depressive disorder (MDD), which were further supported by animal behavioral tests. Our findings demonstrate that gut microbiota mainly modulate the mutual transformation of microglial subtypes, which may lead to new insights into the pathogenesis of AD and MDD.

Carbon monoxide poisoning with hippocampi lesions on MRI: cases report and literature review.

BACKGROUND: Carbon monoxide (CO) poisoning is now one of the leading causes of poisoning-related mortality worldwide. The central nervous system is the most vulnerable structure in acute CO poisoning. MRI is of great significance in the diagnosis and prognosis of CO toxic encephalopathy. The imaging features of CO poisoning are diverse. We report atypical hippocampal lesions observed on MRI in four patients after acute CO exposure. CASE PRESENTATIONS: We report four patients who presented to the emergency department with loss of consciousness. The diagnosis of CO poisoning was confirmed on the basis of their detailed history, physical examination and laboratory tests. Brain MRI in all of these patients revealed abnormal signal intensity in hippocampi bilaterally. They all received hyperbaric oxygen therapy. The prognosis of all four patients was poor. CONCLUSION: Hippocampi, as a relatively rare lesion on MRI of CO poisoning, is of important significance both in the early and delayed stages of acute CO poisoning. In this article, we summarize the case reports of hippocampal lesions on MRI in patients with CO poisoning in recent years, in order to provide reference for the diagnosis and prognosis of CO poisoning.

First Report of Colletotrichum fioriniae Causing Anthracnose on Persimmon in China.

Persimmons (Diospyros kaki Thunb.) have a longstanding history of cultivation in China. Both aesthetically pleasing and edible, they often symbolize a sweet and fulfilling life. During the summer of 2022, a severe outbreak of anthracnose was observed on the lower leaves of persimmon trees in the National Field Genebank for Persimmon (NFGP), located in Yangling, Shaanxi, China (34°17'42.80″ N, 108°04'08.21″ E). The estimated incidence rate of this disease within the NFGP was approximately 30%. The typical symptoms of the disease included the presence of irregular lesions on leaves, and oval sunken lesions on infected fruit. Under high humidity conditions, pink sticky substances appeared in the affected areas. The presence of numerous lesions led to softening and detachment of persimmon fruit. To identify the causal pathogen, 5 × 5mm samples of the diseased leaves were collected from the interface between the infected and healthy leaves. The leaves were disinfected with 70% alcohol for 20 s, followed by rinsing with sterile water. Subsequently, the leaves were immersed in 1% NaClO for 2 to 3 minutes, rinsed with sterile water three times, dried using sterile absorbent paper, and the leaf samples were then transferred onto potato dextrose agar (PDA) medium, and cultured in 25°C incubators. Once the colony reached a certain size, small pieces of hyphae were extracted from edge and transferred for purification and repeated three times. After being cultured on PDA for 7 days, the colony showed a white spongy surface with a pink-orange center. The conidia displayed a fusiform shape and were transparent, measuring 4.58 to 6.53 µm × 9.27 to 13.11 µm (n=50). The isolates share morphological similarities with Colletotrichum fioriniae. The representative isolate HY-7 was selected for molecular identification. The internal transcribed spacers (ITS) region, chitin synthase (CHS-1), actin (ACT), beta-tubulin 2 (TUB2), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene were amplified using ITS1/4 (White et al. 1990), CHS-79F/CHS-345R (Carbone & Kohn, 1999), ACT512F/ACT (Carbone & Kohn, 1999), T1/BT2B (Glass & Donaldson 1995, O'Donnell et al., 1997), and GDF/GDR (Templeton et al. 1992), respectively. The generated sequences were deposited at GenBank under accession numbers OR878056 (ITS), OR766019 (CHS-1), OR766021(TUB2), OR766018 (ACT) and OR766020 (GAPDH). BLAST analysis revealed the sequences were 100% identical to C. fioriniae (MH865005 for ITS, JQ948953 for CHS-1, JQ949613 for ACT, JQ949943 for TUB2 and JQ948622 for GAPDH). The morphological characteristics and molecular analyses of the isolate matched the description of C. fioriniae. To fulfill Koch's postulates, the twigs and leaves of 'Fupingjianshi' in four different directions were inoculated without wounding in the field, and 10 healthy fruits were selected for wound inoculation. The concentration of conidia used for inoculation was about 1 × 106 conidia/ml, and sterilized water was used as control. The experiment was replicated three times under the same conditions. One week after inoculation, characteristic symptoms resembling those observed on the leaves of primary diseased persimmon trees appeared on the leaves and fruits. No symptoms were observed on the leaves, twigs and fruits in the control treatment. The pathogen from the artificially infected leaves and fruits were reisolated and identified as C. fiorinae based on morphological and molecular characteristics. Persimmon anthracnose is a common disease in regions where the fruit is grown, to the best of our knowledge, this is the first documented occurrence of C. fioriniae-induced anthracnose on persimmons in China, which should be paid more attentions. This report will help identify disease symptoms in the field and provides a basis for determining the occurrence, distribution, and control of C. fioriniae on persimmon leaves and fruits.

Evaluating the efficacy of standardized pressure ulcer management protocols in the prevention of pressure injuries among patients undergoing neurosurgical procedures.

Pressure injuries are a significant concern for patients undergoing neurosurgical procedures due to prolonged immobility and the complexity of care. This study evaluates the efficacy of standardized pressure ulcer management protocols in preventing pressure injuries and enhancing patient care in a neurosurgical context. A comprehensive retrospective analysis was conducted at a single institution from December 2020 to December 2023, comparing 50 patients who received standardized pressure ulcer management (intervention group) with 50 patients who received conventional care (control group). The study assessed the incidence of pressure ulcers, patient comfort levels using the Kolcaba Comfort Scale and sleep quality using the Richards-Campbell Sleep Questionnaire (RCSQ). Statistical analysis was performed using SPSS software, version 27.0, applying t-tests and chi-square tests as appropriate. The intervention group exhibited a significantly lower incidence of pressure ulcers at all measured time points post-surgery compared to the control group. Patient comfort levels in the intervention group were consistently higher across psychological, environmental, physiological and socio-cultural domains. Sleep quality metrics, including sleep depth, latency to sleep onset and overall sleep quality, were significantly improved in the intervention group. The implementation of standardized pressure ulcer management protocols in neurosurgical care significantly reduces the incidence of pressure injuries, enhances patient comfort and improves sleep quality. These findings highlight the importance of adopting structured care protocols to improve postoperative outcomes and patient well-being in neurosurgical settings.

Genome-wide analysis of circular RNA-mediated ceRNA regulation in porcine skeletal muscle development.

BACKGROUND: As a diverse and abundant class of endogenous RNAs, circular RNAs (circRNAs) participate in various biological processes including cell proliferation and apoptosis. Nevertheless, few researchers have investigated the role of circRNAs in muscle development in cultivated pigs. RESULTS: In this study, we used RNA-seq to construct circRNA expression profiles in skeletal muscle of Jinfen White pigs at the age of 1, 90, and 180 days. Among the 16,990 identified circRNAs, 584 circRNAs were differentially expressed. Moreover, the enrichment analysis of DE circRNA host genes showed that they were mainly involved in muscle contraction, muscle organ development and muscle system processes, as well as AMPK and cAMP-related signal pathways. We also constructed a circRNA-miRNA-mRNA co-expression network to find key circRNAs which many involved in the regulation of porcine skeletal muscle development through the competitive endogenous RNA (ceRNA) mechanism. It is noteworthy that circ_0018595/miR-1343/PGM1 axis may play a regulatory role in the development of porcine skeletal muscle. CONCLUSIONS: This study identified the circRNAs and present the circRNA expression profile in the development of pigs, revealed that DE circRNA host genes participate in different cell fates and enriched the porcine ceRNA network. Thus, this work will become a valuable resource for further in-depth study of the regulatory mechanism of circRNA in the development of porcine skeletal muscle.

Hesperidin inhibits methylation and autophagy in LPS and high glucose-induced human villous trophoblasts.

INTRODUCTION: Gestational diabetes mellitus (GDM) is the first occurrence of diabetes due to abnormal maternal sugar metabolism after pregnancy, which may lead to adverse pregnancy outcomes. Hesperidin is known to decrease in the cord blood of GDM with obesity, but its role is unknown. This study aims to explore the potential function of hesperidin in GDM with obesity to develop new therapeutic ideas. METHODS: Peripheral blood and placental tissues from GDM and GDM with obesity patients were collected to isolate human villous trophoblasts and detection. Bioinformatics was used to analyze the differential methylation genes between GDM and GDM with obesity. Immunofluorescence was applied for the detection of CK7 expression. Cells vitality was detected by CCK8 and transwell. Molecular docking was applied to predict the binding of hesperidin and ATG7 protein. Inflammation and m6A levels was analyzed by ELISA. ATG7, LC3, TLR4 and P62 proteins was analyzed by Western blot. RESULTS: The methylation of ATG7 gene was up-regulated in GDM with obesity compared with GDM. The m6A and autophagy proteins levels in GDM with obesity were higher than that in GDM. LPS with 2.5-25 mM glucose induced the increase of autophagy proteins, inflammation and m6A levels in human villous trophoblasts. Hesperidin formed hydrogen bonds and hydrophobic interactions with ATG7 proteins. Hesperidin (0.25 µM) inhibited the autophagy proteins and m6A level in LPS and 25 mM glucose-induced human villous trophoblasts. DISCUSSION: GDM with obesity followed the increase of autophagy proteins and m6A levels. Hesperidin inhibited the autophagy proteins and m6A level in LPS and glucose-induced human villous trophoblasts.

Andrographolide induced ferroptosis in multiple myeloma cells by regulating the P38/Nrf2/HO-1 pathway.

Andrographis paniculata is used as a functional food in Asia. Andrographolide (Andro), a diterpene lactone isolated from Andrographis paniculata, has been reported to have potent anticancer activity. Multiple myeloma (MM), the second most common malignant tumor in hematology, is incurable. Ferroptosis, a type of cell death driven by iron-dependent lipid peroxidation, has shown potential in the treatment of various cancers. However, previous studies have not demonstrated whether Andro inhibits the development of MM via ferroptosis or any other mechanism. In the present study, we observed that Andro induced cell death, G0/G1 cell cycle arrest and evoked oxidative stress in MM cells. Interestingly, these phenomena were accompanied by increases in intracellular and mitochondrial Fe2+ and lipid peroxidation levels. Furthermore, treatment with ferroptosis inhibitors rescued Andro-induced cell death, which indicated that ferroptosis contributed to this phenomenon. Mechanistic examination showed that Andro may block the Nrf2/HO-1 signaling pathway by activating P38, thereby inducing ferroptosis. Moreover, inhibition of P38 expression rescued Andro-induced cell death, changes in the level of Nrf2 and HO-1 expression, Fe2+ and lipid peroxidation. Taken together, our findings suggest that Andro induces ferroptosis in MM cells via the P38/Nrf2/HO-1 pathway, providing a potential preventative and therapeutic approach for MM.

Major pathological remissions in a patient with stage IIIA nonsmall cell lung cancer after neoadjuvant tislelizumab combined with chemotherapy: a case report and literature review.

INTRODUCTION: Currently, there are few reports of patients with locally advanced lung cancer achieving a clinical complete response by medical treatment. Preoperative neoadjuvant immunotherapy combined with chemotherapy is an option for patients with unresectable, locally advanced nonsmall cell lung cancer (NSCLC) which is of great potential, and may change traditional treatment paradigms. There are relatively few large-scale, high-quality randomized-controlled trials yet, and limitations such as short postoperative follow-up period and immature disease-free survival and overall survival data still persist. Thus, evidence-based medical evidence is urgently needed. It is worthy to explore the further treatment of patients who achieved complete response after initial treatment, though lacking of evidence by now. CASE PRESENTATION: We report a stage IIIA lung squamous cell carcinoma case who achieved a major pathologic remission after neoadjuvant treatment with tislelizumab and chemotherapy. CONCLUSION: Our case study contributes to the existing evidence on the feasibility, efficacy and safety of neoadjuvant immunotherapy combined with chemotherapy in locally advanced unresectable NSCLC.

Transcriptome Analysis Revealed Potential Genes of Skeletal Muscle Thermogenesis in Mashen Pigs and Large White Pigs under Cold Stress.

Pigs are susceptible to cold stress due to the absence of brown fat caused by the partial deletion of uncoupling protein 1 during their evolution. Some local pig breeds in China exhibit potential cold adaptability, but research has primarily focused on fat and intestinal tissues. Skeletal muscle plays a key role in adaptive thermogenesis in mammals, yet the molecular mechanism of cold adaptation in porcine skeletal muscle remains poorly understood. This study investigated the cold adaptability of two pig breeds, Mashen pigs (MS) and Large White pigs (LW), in a four-day cold (4 °C) or normal temperature (25 °C) environment. We recorded phenotypic changes and collected blood and longissimus dorsi muscle for transcriptome sequencing. Finally, the PRSS8 gene was randomly selected for functional exploration in porcine skeletal muscle satellite cells. A decrease in body temperature and body weight in both LW and MS pigs under cold stress, accompanied by increased shivering frequency and respiratory frequency, were observed. However, the MS pigs demonstrated stable physiological homeostasis, indicating a certain level of cold adaptability. The LW pigs primarily responded to cold stress by regulating their heat production and glycolipid energy metabolism. The MS pigs exhibited a distinct response to cold stress, involving the regulation of heat production, energy metabolism pathways, and robust mitochondrial activity, as well as a stronger immune response. Furthermore, the functional exploration of PRSS8 in porcine skeletal muscle satellite cells revealed that it affected cellular energy metabolism and thermogenesis by regulating ERK phosphorylation. These findings shed light on the diverse transcriptional responses of skeletal muscle in LW and MS pigs under cold stress, offering valuable insights into the molecular mechanisms underlying cold adaptation in pigs.

A predictive nomogram for a failed trial of labor after cesarean: A retrospective cohort study.

AIM: To validate risk factors and a nomogram prediction model for the failure of a trial of labor after cesarean section (TOLAC) in a Chinese population. METHODS: We included women who tried TOLAC between January 2017 and May 2019, grouped according to the success/failure of TOLAC. The patients were randomized 3:1 into the development and validation sets. Multivariable logistic regression analyses were used to develop a nomogram prediction model for TOLAC failure. RESULTS: In total, 535 (86.3%) of the women (n = 620) aged 29-34 years had a successful vaginal birth after cesarean (VBAC). All women had a fully healed previous uterine incision. The univariable analyses showed that the cephalopelvic score (p < 0.001), BMI (p = 0.001), full engagement into the pelvis (p < 0.001), Bishop cervical maturity score (p < 0.001), and estimated fetal weight at admission (p < 0.001) could enter the multivariable model. Furthermore, the multivariable analysis showed that the cephalopelvic score (OR = 0.42, 95%CI: 0.23-0.77, p = 0.005), full engagement in the pelvis (OR = 0.16, 95%CI: 0.08-0.33, p < 0.001), and Bishop cervical maturity score (OR = 0.46, 95%CI: 0.35-0.59, p < 0.001) were independent predictors of the failure of TOLAC. CONCLUSION: This study proposes a nomogram that can assess the risk of failure of TOLAC in Chinese pregnant women. The statistical model could help clinicians know the likelihood of successful TOLAC in the clinical setting.

Fine mapping QTL and mining genes for protein content in soybean by the combination of linkage and association analysis.

Soybean is one main source of dietary protein; therefore, improving protein content is an important objective in breeding programs. There is a significant negative correlation between protein and oil content, which influenced mapping quantitative trait locus (QTL) and quantitative trait nucleotides for these two traits. In this study, a linkage map was created with 2232 single-nucleotide polymorphism markers for the four-way recombinant inbred line (FW-RIL) population derived from the cross (Kenfeng 14 × Kenfeng 15) × (Heinong 48 × Kenfeng 19), and then conditional and unconditional QTL analyses were carried out by inclusive complete interval mapping based on the phenotypic data of protein and oil content collected in 10 different environments. As shown in the results of linkage analysis, a total of 85 QTL have been detected. We have performed association analysis using 109,676 markers after quality filtering for FW-RIL, and the results have shown that a total of 60 QTNs were detected. We have performed association analysis using 63,306 markers after quality filtering for resource population, and the results have shown that a total of 123 QTNs were detected. We have combined linkage and association analysis, and there are six QTNs verified by FW-RIL and resource population. We have performed pathway analysis on the genes in these six QTN attenuation regions, and the result shows that a total of four candidate genes are related to the synthesis or metabolism of soybean protein. These findings will facilitate marker-assisted selection and molecular breeding of soybean.

Mapping QTL/QTN and mining candidate genes for plant height and its response to planting densities in soybean [Glycine max (L.) Merr.] through a FW-RIL population.

Plant height (PH) determines the morphology and seed yield of soybean, so it is an important breeding target, which is controlled by multiple genes and affected by plant density. In this research, it was used about a four-way recombinant inbred lines (FW-RIL) with 144 families constructed by double cross (Kenfeng 14 × Kenfeng 15) × (Heinong 48 × Kenfeng 19) as experimental materials, with the purpose to map QTL/QTN associated with PH under densities of 2.2×105 plant/ha (D1) and 3×105 plant/ha (D2) in five environments. The results showed that response of PH to densities varied in accordance to genotypes among environments. A total of 26 QTLs and 13 QTNs were identified specifically in D1; 20 QTLs and 21 QTNs were identified specifically in D2. Nine QTLs and one QTN were discovered commonly in two densities. Fifteen QTLs and 9 QTNs were repeatedly detected by multiple statistical methods, densities, or environments, which could be considered stable. Eighteen QTLs were detected, as well as 7 QTNs underlying responses of PH to density increment. Six QTNs, co-located in the interval of QTL, were detected in more than two environments or methods with a longer genome length over 3000 kb. Based on gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, five genes were predicted as candidates, which were likely to be involved in growth and development of PH. The results will help elucidate the genetic basis and improve molecular assistant selection of PH. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01209-0.

Prognostic value of preoperative lymphocyte-related systemic inflammatory biomarkers in upper tract urothelial carcinoma patients treated with radical nephroureterectomy: a systematic review and meta-analysis.

BACKGROUND: Growing evidence shows that the preoperative lymphocyte-related systemic inflammatory biomarkers are associated with the prognosis of patients with upper tract urothelial carcinoma (UTUC). These markers include neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and monocyte-to-lymphocyte ratio (MLR). However, these findings are inconsistent, and the prognostic significance of these biomarkers is unclear. Moreover, the currently available prognostic indicators do not precisely predict the outcome of UTUC patients. This motivated us to investigate the prognostic values of NLR, PLR, and MLR in UTUC patients treated with radical nephroureterectomy (RNU). METHODS: We prospectively registered this in PROSPERO (CRD42020186531). We performed a comprehensive literature search of the PubMed, Web of Science, EMBASE, and Cochrane Library databases to identify the eligible studies evaluating the prognostic values of preoperative NLR, PLR, and MLR. Hazard ratios with 95% confidence intervals of overall survival (OS), cancer-specific survival (CSS), disease-free survival (DFS), recurrence-free survival (RFS), metastasis-free survival (MFS), and progression-free survival (PFS) were extracted from the multivariate analyses and analyzed with fixed or random effects models when applicable. Heterogeneity among the studies was evaluated using Cochran's Q test and I2 statistic. Sensitivity and subgroup analyses were conducted to explore the origin of heterogeneity. The Newcastle-Ottawa Scale (NOS) was applied to assess the quality of each enrolled study. Publication bias was determined using funnel plots together with Egger's tests. The Grading of Recommendations Assessment, Development, and Evaluation (GRADE) was used to evaluate the quality of the evidence. RESULTS: Overall, we included 10,339 UTUC patients from twenty-five retrospective studies. The results indicated that elevated preoperative NLR, PLR, and MLR were significantly associated with worse OS, CSS, DFS/RFS/MFS, and PFS in the UTUC patients undergoing RNU. Furthermore, the results of sensitivity and subgroup analyses demonstrated the rationality and reliability of the results. CONCLUSIONS: The present meta-analysis demonstrated a significant association between elevated preoperative NLR, PLR, and MLR and poor prognosis in patients with surgically treated UTUC. Hence, lymphocyte-related systemic inflammatory biomarkers, in conjunction with clinicopathological factors, molecular markers, and other prognostic indicators, could be helpful to determine the primary treatment strategies and to design individualized follow-up plans for UTUC patients.

Hypermethylation of dihydrofolate reductase promoter increases the risk of hypertension in Chinese.

BACKGROUND: DNA methylation was considered to play an important role in hypertension. However, the direct association between dihydrofolate reductase (DHFR) promoter methylation and hypertension remains unclear. We thus aimed to investigate the relationship between DNA methylation of DHFR promoter and hypertension. MATERIALS AND METHODS: A total of 371 hypertensive patients (diastolic blood pressure ≥90 mmHg and/or systolic blood pressure ≥140 mmHg or a history of antihypertensive treatment) and 320 age- and sex-matched healthy controls from the Hypertension Management Information System in Nanshan Community Health Service Centers were included in this case-control study. Quantitative methylation-specific polymerase chain reaction was used to measure the level of DHFR promoter methylation, which was presented as the percentage of methylated reference (PMR). A multivariate logistic regression model was used to explore the risk of DHFR promoter methylation. RESULTS: Our results indicated that the level of DHFR promoter methylation was higher in hypertensive patients (median PMR, 34.32%; interquartile range, 11.34-119.60) than in healthy controls (median PMR, 18.45%; interquartile range, 8.16-35.40) (P < 0.001). Multivariable analysis showed that the risk of DHFR promoter hypermethylation was significantly higher in hypertensive patients than in healthy controls (odds ratio = 3.94, 95% confidence interval = 2.56-6.02, P < 0.001). Furthermore, hypermethylation was positively associated with sex, high blood homocysteine levels, and alcohol drinking. In particular, the area under the receiver operating characteristic curve was 0.688 (0.585-0.668) for the male hypertensive patients, suggesting the potential diagnostic value of DHFR promoter methylation in male hypertension. CONCLUSION: Our results demonstrated that DHFR promoter hypermethylation is positively associated with the risk of hypertension in Chinese.

Silencing lncRNA ZFAS1 or elevated microRNA-135a represses proliferation, migration, invasion and resistance to apoptosis of osteosarcoma cells.

BACKGROUND: Osteosarcoma (OS) is still a disease with high mortality from malignant tumors in children and adolescents. Due to its poor treatment, this study explored the involvement of lncRNA ZFAS1/microRNA-135a (miR-135a)/apurinic/apyrimidinic exonuclease 1 (APEX1) axis in the regulation of OS growth and metastasis. METHODS: ZFAS1, miR-135a and APEX1 expression in OS tissues and cells were tested by RT-qPCR and western blot analysis. MG63 cells were transfected with sh-ZFAS1, miR-135a mimic or their controls to unearth theirs functions in the proliferation, colony formation, migration, invasion, cycle entry and apoptosis of MG63 cells by MTT and EdU, colony formation assays, flow cytometry, and Transwell assay, severally. The proliferation related factor (Ki-67, CyclinD1), apoptosis related factor (Bax, Bcl-2) and migration related factor (MMP2, MMP9) protein levels were tested. Tumor volume and weight were detected by subcutaneous tumor xenograft in nude mice. RESULTS: Overexpressed ZFAS1 and APEX1, and down-regulated miR-135a existed in OS tissues and cells. Silenced ZFAS1 or elevated miR-135a inhibited colony formation and proliferation, cycle progression, migration and invasion while promoted apoptosis of MG63 cells. Silenced ZFAS1 or elevated miR-135a suppressed tumor volume and weight of OS in vivo. LncRNA ZFAS1 promoted APEX1 expression by competitively binding with miR-135a. CONCLUSION: This study indicates that silenced ZFAS1 or up-regulated miR-135a restrained migration, proliferation and invasion and promoted apoptosis of OS MG63 cells. This study provides a possible theoretical basis for studying the regulatory mechanism of ZFAS1/miR-135a/APEX1 signaling axis on the growth and metastasis of OS.

Rip2 Is Required for Nod2-Mediated Lysozyme Sorting in Paneth Cells.

Paneth cells play an important role in maintaining intestinal homeostasis by secreting a large number of antimicrobial peptides into the intestinal lumen. In this study, we found that Rip2 is required for lysozyme sorting in Paneth cells in a manner that is dependent on Nod2, LRRK2, and Rab2a. Rip2 deficiency in mouse led to lysosomal degradation of lysozyme in Paneth cells and prevented the recruitment of Rab2a onto dense core vesicles (DCVs). Like Nod2 and LRRK2, Rip2 localizes to DCVs in Paneth cells, and its DCV localization depends on Nod2 and LRRK2. Thus, we delineated a genetic pathway, consisting of Nod2-LRRK2-Rip2-Rab2a, which is required for lysozyme sorting. Taken together, our results indicate that the lysozyme-sorting process in Paneth cells is orchestrated by a number of host factors and highlight the importance of Paneth cell function in intestinal homeostasis.

2Ch-2N square and hexagon interactions: a combined crystallographic data analysis and computational study.

In recent years, chalcogen bonding (ChB), a typical σ-hole interaction, has shown great potential as a bottom-up design approach for specific applications. According to our survey of the Cambridge Structural Database (CSD), a large number of crystal structures containing 2Ch-2N square and hexagon interaction motifs were extracted. On the basis of the CSD search results, the 2Ch-2N square interactions in the dimers of 2,1,3-benzochalcogenadiazole 1 and 2,1,3-pyridochalcogenadiazole 2, together with 2Ch-2N hexagon interactions in the dimers of chalcogenazolo-pyridine 3 and triazolo-chalcogenadiazole 4, were firstly studied. Then, substituent effects on these peculiar interactions were thoroughly examined by introducing a diversity of small, non-aromatic substituents at the 4,7-positions of the 1S scaffold and various aryl substituents at the 2-position of the 3Te scaffold. Our calculations showed that the major contribution to the attraction of such bidentate ChB interactions is electrostatics, while the orbital term also plays an important role. Some strong electron-withdrawing substituents, such as NO2, CN, and CF3, tend to enhance square ChB interactions, while C6F5 and CF3 substituents with a strong electron-withdrawing ability strengthen hexagon ChB interactions. Particularly, a good linear correlation has been established between the binding energies of the dimers under study and both the surface electrostatic potential (ESP) maxima for the Ch σ-holes and the minimum surface ESP of the N atoms, which provides reasonable models to evaluate these interactions. The results reported in this work will provide design guidance for the applications of 2Ch-2N cyclic motifs in materials science and biochemistry.

HDM2 Promotes NEDDylation of Hepatitis B Virus HBx To Enhance Its Stability and Function.

Hepatitis B virus (HBV)-encoded X protein (HBx) plays a critical role in HBV-related hepatocarcinoma development. In this study, we demonstrate that HBx is specifically modified by NEDD8. We found that E3 ligase HDM2 promotes NEDDylation of HBx to enhance HBx stability by preventing its ubiquitination-mediated degradation. Consistently, analysis of 160 hepatocellular carcinoma patient specimens indicated that the amount of HDM2 protein correlates with HBx protein level. We identified that HBx K91 and K95 as the key HBx NEDDylation sites and observed that the NEDDylation-deficient HBx has shorter half-life. We generated Huh7 cell lines which ectopically express wild-type and NEDDylation-deficient HBx and found that NEDDylation-deficient HBx showed less chromatin localization and less DDB1 binding. Consistently, the expression of HBx-regulated genes (IL-8, MMP9, and YAP) and HBV transcription (the activity of HBV enhancer and the amount of pgRNA transcribed from cccDNA) were significantly higher in cells expressing wild-type (WT) HBx than that in cells expressing mutant HBx. In addition, HBx-expressing cells proliferated faster than control and mutant HBx-expressing cells. We also showed that the ability of WT HBx-expressing cells to form tumors in nude mice was significantly higher than that of mutant HBx-expressing cells. In conclusion, we revealed that E3 ligase HDM2 promotes NEDDylation of HBx to enhance HBx stability and chromatin localization, which in turn favors HBx-dependent transcriptional regulation, cell proliferation, and HBV-driven tumor growth.IMPORTANCE Hepatitis B virus (HBV) HBx protein plays a critical role in viral replication and hepatocarcinogenesis. However, the regulation of HBx stability is not well understood. We found that HBx is modified by NEDD8 and that the HDM2 E3 ligase promotes HBx NEDDylation to enhance HBx stability by inhibiting its ubiquitination. We provide a new evidence to show the positive correlation between HDM2 and HBx in clinical hepatocellular carcinoma (HCC) samples. We also identified the major NEDDylation sites on HBx. Our studies indicate that the defective NEDDylation of HBx negatively affects its ability to activate the transcription of downstream genes and promote cell proliferation and tumor growth in vivo Taken together, our findings reveal a novel posttranslational modification of HBx by HDM2 which regulates its stability, subcellular localization, and functions. These findings indicate that HDM2 is an important regulator on HBx and a potential diagnosis/therapeutic marker for HBV-associated HCC.