Study on the Adsorption Behavior of Polymeric Dispersants to S-ZnF Particles during Grinding Process.

Three sodium polyacrylate copolymers PD0x (Poly acrylic acid-co-sodium 4-vinylbenzenesulfonate or PD01; Poly acrylic acid-co-sodium 4-vinylbenzenesulfonate-co-hydroxyethyl methacrylate or PD02 and Poly methyl methacrylate-co-acrylic acid-co-sodium 4-vinylbenzenesulfonate-co-hydroxyethyl methacrylate or PD03) were synthesized as water-based dispersants for grinding red-brown pigment ZnFe1.2Cr0.8O4 particles prepared by the solid phase method (S-ZnF). The particle size distribution, viscosity of suspensions, and adsorption capacity of dispersants were explored by laser particle size analysis, viscometer, and thermogravimetry (TG), respectively. The application of 2 wt.% dispersant PD02 in the S-ZnF suspension ground for 90 min can deliver a finer product with the narrower particle size distribution. The added dispersant PD02 in the grinding process of the S-ZnF particles exhibits a suitable viscosity of the suspension and generates more hydrogen bonds on the S-ZnF particle surface. The sulfonic acid groups (SO3-) and carboxylic acid groups (-COO-) in the dispersant PD02 can also provide a strong charge density, which is favorable for the dispersion and grinding of the S-ZnF particles in the suspensions. Furthermore, the adsorption behavior of polymeric dispersant PD02 adsorbed on the S-ZnF particles surface was simulated and analyzed by adsorption thermodynamic models and adsorption kinetic models. It is indicated that the adsorption thermodynamic behavior of dispersant PD02 adsorbed on the S-ZnF particles surface follows the Langmuir model, and the adsorption process is endothermic and a random process with increased confusion during the grinding process. In addition, the adsorption kinetics of dispersant PD02 adsorbed on the S-ZnF particles surface are more in line with the pseudo-first-order kinetic models. Therefore, the adsorption process of dispersant PD02 on the S-ZnF particles surface can be considered as a single-surface adsorption process.

A New Class of Proton Conductors with Dramatically Enhanced Stability and High Conductivity for Reversible Solid Oxide Cells.

Reversible solid oxide cells based on proton conductors (P-ReSOCs) have potential to be the most efficient and low-cost option for large-scale energy storage and power generation, holding promise as an enabler for the implementation of intermittent renewable energy technologies and the widespread utilization of hydrogen. Here, the rational design of a new class of hexavalent Mo/W-doped proton-conducting electrolytes with excellent durability while maintaining high conductivity is reported. Specifically, BaMo(W)0.03 Ce0.71 Yb0.26 O3-δ exhibits dramatically enhanced chemical stability against high concentrations of steam and carbon dioxide than the state-of-the-art electrolyte materials while retaining similar ionic conductivity. In addition, P-ReSOCs based on BaW0.03 Ce0.71 Yb0.26 O3-δ demonstrate high peak power densities of 1.54, 1.03, 0.72, and 0.48 W cm-2 at 650, 600, 550, and 500 °C, respectively, in the fuel cell mode. During steam electrolysis, a high current density of 2.28 A cm-2 is achieved at a cell voltage of 1.3 V at 600 °C, and the electrolysis cell can operate stably with no noticeable degradation when exposed to high humidity of 30% H2 O at -0.5 A cm-2 and 600 °C for over 300 h. Overall, this work demonstrates the promise of donor doping for obtaining proton conductors with both high conductivity and chemical stability for P-ReSOCs.

CCND2 and miR-206 as potential biomarkers in the clinical diagnosis of thyroid carcinoma by fine-needle aspiration cytology.

BACKGROUND: To investigate the relationship between cyclin D2 (CCND2) and miR-206 expression in fine-needle aspiration cytology of thyroid carcinoma. METHODS: A total of 65 patients with thyroid carcinoma were selected as the subjects and 65 patients with benign thyroid nodules were in control group. The fine-needle aspiration cytology of thyroid nodules was performed. CCND2 and miR-206 levels were detected by PCR. RESULTS: Compared with the patients with benign thyroid nodules, the expression level of miR-206 in fine-needle aspiration cytology of thyroid cancer patients decreased significantly and the expression level of CCND2 increased significantly. CCND2 and miR-206 expression was negatively correlated in thyroid cancer tissues. Area under curve (AUC) of miR-206 level in the diagnosis of thyroid cancer was 0.889, and the sensitivity and specificity were 92.3% and 81.5%, respectively. AUC of CCND2 level in the diagnosis of thyroid cancer was 0.837, and the sensitivity and specificity were 67.7% and 89.2%, respectively. The AUC of combined detection of CCND2 and miR-206 in the diagnosis of thyroid cancer was 0.959, and the sensitivity and specificity were 93.8% and 87.7%, respectively. The levels of miR-206 and CCND2 were significantly correlated with TNM staging and lymph node metastasis. CONCLUSIONS: miR-206 and CCND2 may become new biomarkers for clinical diagnosis of thyroid cancer based on the fine-needle aspiration cytology of thyroid nodules.

The Unique ergodic stationary distribution of two stochastic SEIVS epidemic models with higher order perturbation.

Two types of susceptible, exposed, infectious, vaccinated/recovered, susceptible (SEIVS) epidemic models with saturation incidence and temporary immunity, driven by higher order white noise and telegraph noise, are investigated. The key aim of this work is to explore and obtain the existence of the unique ergodic stationary distribution for the above two models, which reveals whether the disease will be prevalent and persistent under some noise intensity assumptions. We also use meticulous numerical examples to validate the feasibility of the analytical findings. Finally, a brief biological discussion shows that the intensities of noises play a significant role in the stationary distributions of the two models.

Crowded Environment Regulates the Coacervation of Biopolymers via Nonspecific Interactions.

The membrane-less organelles (MLOs) with subcompartments are formed via liquid-liquid phase separation (LLPS) in the crowded cell interior whose background molecules are up to 400 mg/mL. It is still a puzzle how the background molecules regulate the formation, dynamics, and functions of MLOs. Using biphasic coacervate droplets formed by poly(l-lysine) (PLL), quaternized dextran (Q-dextran), and single-stranded oligonucleotides (ss-oligo) as a model of MLO, we online monitored the LLPS process in Bovine Serine Albumin (BSA) solution up to 200.0 mg/mL. Negatively charged BSA is able to form complex or coacervate with positively charged PLL and Q-dextran and thus participates in the LLPS via nonspecific interactions. Results show that BSA effectively regulates the LLPS by controlling the phase distribution, morphologies, and kinetics. With increasing BSA concentration, the spherical biphasic droplets evolve in sequence into phase-inverted flower-like structure, worm-like chains, network structures, and confined coacervates. Each kind of morphology is formed via its own specific growth and fusion pathway. Our work suggests that MLOs could be controlled solely by the crowded environment and provides a further step toward understanding the life process in cell.

The prognostic impact of tumor-infiltrating B lymphocytes in patients with solid malignancies: A systematic review and meta-analysis.

This study reviewed the prognostic effect of tumor-infiltrating B lymphocytes (TIBLs) on solid malignancies, to determine the potential role of TIBLs in predicting cancer patient's prognosis and their response to immunotherapy. A total of 45 original papers involving 11,099 individual patients were included in this meta-analysis covering 7 kinds of cancer. The pooled results suggested that high levels of TIBLs were correlated with favorable OS in lung, esophageal, gastric, colorectal, liver, and breast cancer; improved RFS in lung cancer; and improved DFS in gastrointestinal neoplasms. Additionally, TIBLs were significantly correlated with negative lymphatic invasion in gastric cancer, small tumor size in hepatocellular carcinoma, and negative distant metastasis in colorectal cancer. Additionally, TIBLs were reported as a discriminative feature of patients treated with immunotherapy with improved survival. We concluded that TIBLs play a favorable prognostic role among the common solid malignancie, providing theoretical evidence for further prognosis prediction for solid tumors.

Length and rigidity of the spacer impact on aldose reductase inhibition of the 5F-like ARIs in a dual-occupied mode.

The primary objective of this study was to investigate the structure-activity relationship of a new series of 5F-like Aldose Reductase Inhibitors (ARIs) using in silico docking method. In this perspective, 6 novel ARIs have been designed and synthesized. Evaluation of the inhibition of these compounds to ALR2 was carried on with epalrestat and 5F as the references. It was found that the spacer of 5F-like ARIs has a great influence on their inhibitory activity. Rigid spacer with length equal to 3 âˆ¼ 4 carbon alkyl chain brings about better inhibitory activity. Among them, compound 4b was verified as the most active ARIs, where its IC50 value was 16.8 ± 1.3 nM. Furthermore, in silico docking studies using AutoDock 4.2 as well as molecular simulation using GROMACS 2022.1 showed that 5F-like ARIs adopt a dual-occupation mode. The interaction energy (-25 to -74 kcal/mol), as well as MM-GBSA binding free energy (-37 to -65 kcal/mol) was positively correlated with their ALR2 inhibition constant (2000 to 16.8 nM). Docking interaction explained well the structure-activity relationship. A pharmacophore model has been set up for 5F-like ARIs thereafter. This model indicates that as an effective ARI, the entity should have four characteristics: an aromatic center, two hydrogen bond donors, and one hydrogen bond acceptor. By the way, all the 5F-like ARIs reported here are good to mild antioxidant with EC50 value between 13.6 ± 1.2 and 71.1 ± 3.2 µM. All our data direct the further development of more optimal ARIs for the treatment of diabetic complication in the future.

Multi-Level Support Technology and Application of Deep Roadway Surrounding Rock in the Suncun Coal Mine, China.

To solve these problems of poor supporting effect and serious deformation and failure of surrounding rock of mining roadway under deep mining stress, a FLAC-3D numerical calculation model is established with -800 m level no. 2424 upper roadway in the Suncun Coal Mine as the background to compare the stress, deformation, and failure law of surrounding rock of mining roadway under once support and multi-level support with the same support strength. It is found that the multi-level support technology has obvious advantages in the surrounding rock of the horizontal roadway on the 2424 working face. From this, the key parameters of multi-level support are determined, and the field industrial test is carried out. The results show that the overall deformation of the surrounding rock is obviously reduced after multi-level support. The displacement of the two sides is reduced by about 40%, the displacement of the roof and floor is reduced by about 30%, and the plastic zone of the roadway is reduced by about 75%. The peak value of concentrated stress decreases from 98.7 MPa to 95.8 MPa, which decreases slightly. The integrity and stability of the surrounding rock are excellent, and the support effect is satisfactory. The research can provide reference and technical support for surrounding rock control of deep high-stress mining roadways.

Symptom clusters and impact on quality of life in esophageal cancer patients.

BACKGROUND: Esophageal cancer patients can experience co-occurring, related symptoms labeled symptom clusters. This study aimed to identify symptom clusters and explore which SCs independently affect the quality of life (QoL) among esophageal cancer patients. METHODS: This cross-sectional study was performed in Shenyang, China, from February 2021 to February 2022. Finally, 118 esophageal cancer patients effectively completed the survey. Questionnaires' information included the Functional Assessment of Cancer Therapy-General (FACT-G), the MD Anderson Symptom Inventory Gastrointestinal Cancer Module (MDASI-GI), the Multidimensions Scale of Perceived Social Support, and demographic and clinical characteristics. Exploratory factor analysis with principal axis factoring was used to identify symptom clusters, and multiple regression analysis was employed to analyze the influencing factors of QoL. RESULTS: The mean score of FACT-G was 69.88 (SD = 17.85) among 118 esophageal cancer patients. Four symptom clusters were identified: psychological-somatic, dysphagia, fatigue-pain, and gastrointestinal symptom clusters. Results of regression analysis indicated a significant impact on QoL for chemotherapy (ß = 0.140, P < 0.045), psychological-somatic symptom cluster (ß = - 0.329, P = 0.013), and social support (ß = 0.409, P < 0.001) after adjusting demographic and clinical characteristics. The linear combination explained 47.8% of the variance in QoL. CONCLUSIONS: There is a critical need to emphasize the importance of psychological-somatic symptoms clusters management programs and increasing social support to improve QoL in esophageal cancer patients.

Transverse testicular ectopia with persistent Mullerian duct syndrome: Report and review of two cases.

Transverse testicular ectopia is a rare anomaly characterized by both testes descending through a single inguinal canal. The objective of this study was to investigate the pathogenesis, diagnosis, and treatment of transverse testicular ectopia (TTE) with persistent Mullerian duct syndrome (PMDS), and to deepen the understanding of the disease in clinical. A retrospective analysis of the clinical manifestation, diagnosis, and treatment of two children suffering from TTE with PMDS was conducted. Previous studies on the characteristics, diagnosis, and treatment of this disease were reviewed. The two patients were treated with laparoscopy-assisted transseptal orchidopexy-inguinal evaluation. After the surgery, the two patients recovered well. The follow-up visits were done 3 months after the operation. An ultrasound examination confirmed that the two patients had testes in the orthotopic position and normal size. TTE with PMDS is an exceedingly rare disease. The patients manifested cryptorchidism on one side; contralateral inguinal hernia was suspected. Detailed physical and ultrasound examinations before the operation are the key to the early diagnosis of TTE. Laparoscopic evaluation is helpful for the diagnosis and finding of other abnormalities. Surgical treatment is the only method to cure the disease; long-term follow-up is needed after TTE operation.

Leveraging Nanodrug Delivery System for Simultaneously Targeting Tumor Cells and M2 Tumor-Associated Macrophages for Efficient Colon Cancer Therapy.

Tumor-associated macrophages (TAMs) widely exist in the solid tumors, which participate in the entire course of tumor development and execute momentous impacts. Therefore, manipulating TAMs has been identified as an expecting strategy with immense potential for cancer therapy. Herein, a nanodrug delivery system was leveraged for simultaneously targeting tumor cells and M2-type TAMs for efficient colon cancer therapy. The broad-spectrum anticancer chemotherapeutic drug doxorubicin (DOX) was hitchhiked in a mannose-modified bovine serum albumin (MAN-BSA) carrier. The DOX@MAN-BSA nanodrug delivery system was verified to possess feasible physical performances for unhindered systemic circulation and active targeting on colon tumors. DOX@MAN-BSA nanoparticles could be preferentially swallowed by colon tumor cells and M2 TAMs through mannose receptor-mediated endocytosis. Further in vivo antitumor therapy in CT26 colon tumor-bearing mice has achieved remarkable suppression efficacy with satisfactory biosafety. Leveraging the nanodrug delivery system for simultaneously targeting tumor cells and M2 TAMs has contributed a feasible strategy to collaboratively repress the malignant tumor cells and the collusive M2 TAMs for efficient cancer therapy.

Mindfulness mediates the relationship between positive parenting and aggression, depression, and suicidal ideation: A longitudinal study in middle school students.

Previous research has indicated that parenting factors affect the risk of maladaptive psychological outcomes (e.g., aggression, depression, or suicidal ideation), and that positive parenting is a prospective risk factor for maladaptive psychological outcomes. However, the mechanisms underlying the relationships between positive parenting, mindfulness, and maladaptive psychological outcomes remain unknown, as do the processes that mediate the effect of positive parenting on maladaptive psychological outcomes in adolescents. The objective of the present study was to investigate the longitudinal relationship between positive parenting, mindfulness, and maladaptive psychological outcomes in middle school students, as well as the mediating effect of mindfulness in the relationships between positive parenting and depression, aggression, and suicidal ideation. In this study, 386 middle school children (aged 12-16) were tested three times over a period of 6 months. Positive parenting was assessed at Time 1, mindfulness at Time 2, and depression, aggression, and suicidal ideation at Time 3. Using structural equation modeling, positive parenting was revealed to be longitudinally associated with mindfulness and negatively associated with maladaptive psychological outcomes. More crucially, mindfulness mediated the relationship between positive parenting and maladaptive psychological outcomes. This research provides important insights into how to effectively decrease adolescent maladaptive psychological outcomes and highlights the importance of teaching mindfulness to youths.

Salvianolic acid A regulates pyroptosis of endothelial cells via directly targeting PKM2 and ameliorates diabetic atherosclerosis.

Rescuing endothelial cells from pyroptotic cell death emerges as a potential therapeutic strategy to combat diabetic atherosclerosis. Salvianolic acid A (SAA) is a major water-soluble phenolic acid in the Salvia miltiorrhiza Bunge, which has been used in traditional Chinese medicine (TCM) and health food products for a long time. This study investigated whether SAA-regulated pyruvate kinase M2 (PKM2) functions to protect endothelial cells. In streptozotocin (STZ)-induced diabetic ApoE-/- mice subjected to a Western diet, SAA attenuated atherosclerotic plaque formation and inhibited pathological changes in the aorta. In addition, SAA significantly prevented NLRP3 inflammasome activation and pyroptosis of endothelial cells in the diabetic atherosclerotic aortic sinus or those exposed to high glucose. Mechanistically, PKM2 was verified to be the main target of SAA. We further revealed that SAA directly interacts with PKM2 at its activator pocket, inhibits phosphorylation of Y105, and hinders the nuclear translocation of PKM2. Also, SAA consistently decreased high glucose-induced overproduction of lactate and partially lactate-dependent phosphorylation of PKR (a regulator of the NLRP3 inflammasome). Further assay on Phenylalanine (PKM2 activity inhibitor) proved that SAA exhibits the function in high glucose-induced pyroptosis of endothelial cells dependently on PKM2 regulation. Furthermore, an assay on c16 (inhibitor of PKR activity) with co-phenylalanine demonstrated that the regulation of the phosphorylated PKR partially drives PKM2-dependent SAA modulation of cell pyroptosis. Therefore, this article reports on the novel function of SAA in the pyroptosis of endothelial cells and diabetic atherosclerosis, which provides important insights into immunometabolism reprogramming that is important for diabetic cardiovascular disease complications therapy.

Iron-Sulfur Clusters: A Key Factor of Regulated Cell Death in Cancer.

Iron-sulfur clusters are ancient cofactors that play crucial roles in myriad cellular functions. Recent studies have shown that iron-sulfur clusters are closely related to the mechanisms of multiple cell death modalities. In addition, numerous previous studies have demonstrated that iron-sulfur clusters play an important role in the development and treatment of cancer. This review first summarizes the close association of iron-sulfur clusters with cell death modalities such as ferroptosis, cuprotosis, PANoptosis, and apoptosis and their potential role in cancer activation and drug resistance. This review hopes to generate new cancer therapy ideas and overcome drug resistance by modulating iron-sulfur clusters.

Association between food intake and mental health problems among preschoolers in China.

Objective: Our knowledge of associations between specific types of food and mental health problems in preschool children, an important developmental period, remains scarce. Using a large sample size and a brief measure, we aimed to examine associations between food groups and mental health problems in preschoolers. Methods: A descriptive cross-sectional study was conducted among 19,548 preschoolers in nine counties in China using a two-stage cluster sampling method. Information on food frequencies, physical activity, sedentary behavior, and sleep conditions was collected by a parent-reported questionnaire. Mental health status was assessed by a parent-reported Strength and Difficulty Questionnaire (SDQ). Height and weight were measured, and BMI was calculated. Relationships between specific types of food groups and mental health status were examined using multivariable regression modeling. Results: Of the present study sample, 47.45% of children were girls. Boys had higher scores relating to total difficulties, conduct problems, hyperactivity, and peer problems than girls but lower scores in emotional symptoms and prosocial behavior. Children with more types of food categories tended to get lower SDQ total scores even after adjusting for confounding variables (p < 0.0001). The subscale scores of SDQ varied depending on the types and numbers of food consumed. Notably, boys' mental health problems were more likely to be associated with their consumed food type. Processed meat frequency intake among boys and girls was positively related to prosocial behavior (Girls: pickled food: p = 0.003, fried food: p < 0.0001, barbecued food: p = 0.006, smoked food: p = 0.005; Boys: pickled food: p = 0.3, fried food: p < 0.0001, barbecued food: p = 0.006, smoked food: p = 0.005) and negatively associated with the SDQ total and the other subscale scores (Girls: pickled food: p ≤ 0.02, fried food: p ≤ 0.005 except for emotions (p = 0.1), barbecued food: p ≤ 0.047, smoked food: p ≤ 0.04 (except for emotions (p = 0.06) and conduct behaviors (p = 0.2)); Boys: pickled food: p < 0.01 except for hyperactivity (p = 0.5), fried food: p ≤ 0.005 except for emotions (p = 0.1), barbecued food: p ≤ 0.007 except for hyperactivity (p = 0.8), smoked food: p ≤ 0.001 except for hyperactivity (p = 0.1) and peer problem (p = 0.08)). Adequate intake frequency of confectionery (2-4 times/week) was beneficial to their emotions, conduct, and total difficulties and was also beneficial for peer and prosocial behavior for boys (Girls: emotions: p = 0.02, conduct: p = 0.02, hyperactivity: p = 0.4, peer: p = 0.06, total difficulties: p = 0.01, prosocial behavior: p = 0.1; Boys: emotions: p = 0.009, conduct: p = 0.0009, hyperactivity: p = 0.7, peer: p = 0.02: total difficulties: p = 0.003, prosocial behavior: p = 0.0002). Conclusion: Diverse food categories and adequate frequency of confectionery were found to be beneficial for children's mental health, with the exception of processed meat. These findings provide novel information and supportive data for children's dietary guidelines and the improvement of mental health.

Guided-mode resonance with reduced bandwidth in mid-infrared absorption and thermal emission.

Narrowband resonance plays an important role in many optical applications, especially for the development of wavelength-selective properties and enhanced light-matter interaction. In this paper, we demonstrate metal-insulator-metal (MIM) waveguide gratings, which exhibit guided-mode resonance (GMR) with reduced bandwidth in mid-infrared absorption and thermal emission. Our fabricated MIM waveguide grating consists of a copper substrate, a lossless ZnSe film, and a top gold stripe grating. Our measurements reveal strong GMRs with a bandwidth of 1.29% of the central wavelength in both mid-infrared absorption and thermal emission spectra. By varying structural parameters of the MIM waveguide grating, strong absorptions and thermal emissions of GMRs are observed and tuned within the 3-5 µm wavelength range. These results manifest the great potential of engineering infrared properties by using GMR and could be useful for spectral control in a variety of infrared devices.

The potential for traditional Chinese therapy in treating sleep disorders caused by COVID-19 through the cholinergic anti-inflammatory pathway.

Since the outbreak of Coronavirus disease (COVID-19) in 2019, it has spread rapidly across the globe. Sleep disorders caused by COVID-19 have become a major concern for COVID-19 patients and recovered patients. So far, there's no effective therapy on this. Traditional Chinese therapy (TCT) has a great effect on sleep disorders, with rare side effects and no obvious withdrawal symptoms. The cholinergic anti-inflammatory pathway, a neuroregulatory pathway in the central nervous system that uses cholinergic neurons and neurotransmitters to suppress inflammatory responses, has been reported to be associated with sleep disorders and psychiatric symptoms. Many studies have shown that TCT activates the cholinergic anti-inflammatory pathway (CAP), inhibits inflammation, and relieves associated symptoms. Therefore, we believe that TCT may be a potential therapeutic strategy to alleviate sleep disorders induced by COVID-19 through CAP. In this review, we analyzed the relationship between cytokine storm induced by Coronavirus and sleep disorders, explained the influence of CAP on sleep disorders, discussed the TCT's effect on CAP, and summarized the treatment effect of TCT on sleep disorders. Based on these practical researches and theoretical basis, we propose potential strategies to effectively improve the sleep disorders caused by COVID-19.

Suicide Gene Delivery System Mediated by Ultrasound-Targeted Microbubble Destruction: A Promising Strategy for Cancer Therapy.

The treatment of malignant tumors has always been one of the challenges that have plagued researchers and clinicians. The ideal status in cancer treatment is to eliminate tumor cells while avoiding damage to normal tissues. Different approaches have been investigated to achieve such a goal, and suicide gene therapy has emerged as a novel mode of cancer treatment. This approach involves the delivery of genes encoding enzymes that activate non-toxic prodrugs into cytotoxic metabolites that cause the death of transfected cancer cells. Despite promising results obtained both in vitro and in vivo, this innovative approach has long been stalled in the clinic due to the lack of a suitable delivery system to introduce the suicide gene into cancer cells. Ultrasound-targeted microbubble destruction (UTMD) represents a valuable non-viral vector system for site-specific and noninvasive gene therapy. Ultrasound promotes intracellular uptake of therapeutic agents by increasing vascular and cell membrane permeability, especially in the presence of microbubbles. In this scenario, the true potential of suicide genes can be translated into clinically valuable treatments for patients. This review provides background information on suicide gene therapy and UTMD technology, summarizes the current state of knowledge about UTMD-mediated suicide gene delivery in cancer treatment, and presents an outlook on its future development.

Novel mutations in GJB1 trigger intracellular aggregation and stress granule formation in X-linked Charcot-Marie-Tooth Disease.

X-linked Charcot-Marie-Tooth Disease type 1(CMT1X) is the second most common form of inherited peripheral neuropathy that is caused by mutations in the gap junction beta-1 (GJB1) gene. Using targeted exome-sequencing, we investigated four CMT families from central-southern China and identified two novel missense variants (p.F31S and p.W44G) and two previously reported variants (p.R220Pfs*23 and p.R164Q) of GJB1. All four probands presented typical early-onset peripheral neuropathy, of which the R220Pfs*23 carrier also had neurologic manifestations in the central nervous system. We then constructed GJB1 expression vectors and performed cell biological analysis in vitro. Expression of FLAG-tagged GJB1 at various time points after transfection revealed evident protein aggregation with both wild-type and mutant forms, indicated with immunostaining and immunoblotting. Detergent-based sequential fractionation confirmed that all mutants were higher expressed and more prone to aggregate than the wild-type, whereas the R220Pfs*23 mutant showed the greatest amount of SDS-soluble multimers and monomers among groups. Moreover, intracellular aggregation probably occurs in the endoplasmic reticulum compartment rather than the Golgi apparatus. Gap junction plaques were present in all groups and were only compromised in frameshift mutant. Further evidence reveals significant intracellular stress granule formation induced by mutated GJB1 and impaired cell viability indicative of cytotoxicity of self-aggregates. Together, our findings demonstrate novel GJB1 variants-induced cell stress and dysfunction and provide insights into understanding the pathomechanisms of GJB1-CMTX1 and other related disorders.

Soil metagenomic analysis on changes of functional genes and microorganisms involved in nitrogen-cycle processes of acidified tea soils.

Nitrogen (N) is the first essential nutrient for tea growth. However, the effect of soil acidification on soil N cycle and N forms in tea plantation are unclear. In this study, the nitrogen contents, soil enzyme activity and N mineralization rate in acidified soil of tea plantation were measured. Moreover, the effects of soil acidification on N cycling functional genes and functional microorganisms were explored by soil metagenomics. The results showed that the NH4 +-N, available N and net N mineralization rate in the acidified tea soil decreased significantly, while the NO3 --N content increased significantly. The activities of sucrase, protease, catalase and polyphenol oxidase in the acidified tea soil decreased significantly. The abundance of genes related to ammonification, dissimilatory N reduction, nitrification and denitrification pathway in the acidified tea soil increased significantly, but the abundance of functional genes related to glutamate synthesis and assimilatory N reduction pathway were opposite. In addition, the abundance of Proteobacteria, Actinobacteria, Chloroflexi, Nitrospirae, Actinomadura, Nitrospira etc. microorganisms related to nitrification, denitrification and pathogenic effect increased significantly in the acidified tea soil. The correlation results showed that soil pH and N forms were correlated with soil enzyme activity, N cycling function genes and microbial changes. In conclusion, soil acidification results in significant changes in enzyme activity, gene abundance and microorganism involved in various N cycle processes in acidified tea soil, which leads to imbalance of soil N form ratio and is not conducive to N transformation and absorption of tea trees.