Guild-level signature of gut microbiome for diabetic kidney disease.

Current microbiome signatures for chronic diseases such as diabetic kidney disease (DKD) are mainly based on low-resolution taxa such as genus or phyla and are often inconsistent among studies. In microbial ecosystems, bacterial functions are strain specific, and taxonomically different bacteria tend to form co-abundance functional groups called guilds. Here, we identified guild-level signatures for DKD by performing in-depth metagenomic sequencing and conducting genome-centric and guild-based analysis on fecal samples from 116 DKD patients and 91 healthy subjects. Redundancy analysis on 1,543 high-quality metagenome-assembled genomes (HQMAGs) identified 54 HQMAGs that were differentially distributed among the young healthy control group, elderly healthy control group, early-stage DKD patients (EDG), and late-stage DKD patients (LDG). Co-abundance network analysis classified the 54 HQMAGs into two guilds. Compared to guild 2, guild 1 contained more short-chain fatty acid biosynthesis genes and fewer genes encoding uremic toxin indole biosynthesis, antibiotic resistance, and virulence factors. Guild indices, derived from the total abundance of guild members and their diversity, delineated DKD patients from healthy subjects and between different severities of DKD. Age-adjusted partial Spearman correlation analysis showed that the guild indices were correlated with DKD disease progression and with risk indicators of poor prognosis. We further validated that the random forest classification model established with the 54 HQMAGs was also applicable for classifying patients with end-stage renal disease and healthy subjects in an independent data set. Therefore, this genome-level, guild-based microbial analysis strategy may identify DKD patients with different severity at an earlier stage to guide clinical interventions. IMPORTANCE: Traditionally, microbiome research has been constrained by the reliance on taxonomic classifications that may not reflect the functional dynamics or the ecological interactions within microbial communities. By transcending these limitations with a genome-centric and guild-based analysis, our study sheds light on the intricate and specific interactions between microbial strains and diabetic kidney disease (DKD). We have unveiled two distinct microbial guilds with opposite influences on host health, which may redefine our understanding of microbial contributions to disease progression. The implications of our findings extend beyond mere association, providing potential pathways for intervention and opening new avenues for patient stratification in clinical settings. This work paves the way for a paradigm shift in microbiome research in DKD and potentially other chronic kidney diseases, from a focus on taxonomy to a more nuanced view of microbial ecology and function that is more closely aligned with clinical outcomes.

Alteration in gut microbiota is associated with immune imbalance in Graves' disease.

Background: Graves' disease (GD), characterized by immune aberration, is associated with gut dysbiosis. Despite the growing interest, substantial evidence detailing the precise impact of gut microbiota on GD's autoimmune processes remains exceedingly rare. Objective: This study was designed to investigate the influence of gut microbiota on immune dysregulation in GD. Methods: It encompassed 52 GD patients and 45 healthy controls (HCs), employing flow cytometry and enzyme-linked immunosorbent assay to examine lymphocyte and cytokine profiles, alongside lipopolysaccharide (LPS) levels. Gut microbiota profiles and metabolic features were assessed using 16S rRNA gene sequencing and targeted metabolomics. Results: Our observations revealed a disturbed B-cell distribution and elevated LPS and pro-inflammatory cytokines in GD patients compared to HCs. Significant differences in gut microbiota composition and a marked deficit in short-chain fatty acid (SCFA)-producing bacteria, including ASV263(Bacteroides), ASV1451(Dialister), and ASV503(Coprococcus), were observed in GD patients. These specific bacteria and SCFAs showed correlations with thyroid autoantibodies, B-cell subsets, and cytokine levels. In vitro studies further showed that LPS notably caused B-cell subsets imbalance, reducing conventional memory B cells while increasing naïve B cells. Additionally, acetate combined with propionate and butyrate showcased immunoregulatory functions, diminishing cytokine production in LPS-stimulated cells. Conclusion: Overall, our results highlight the role of gut dysbiosis in contributing to immune dysregulation in GD by affecting lymphocyte status and cytokine production.

Identification of potential SLA-I-specific T-cell epitopes within the structural proteins of porcine deltacoronavirus.

Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus that mainly threatens newborn piglets and poses a potential broad cross-species transmission risk. The antigenic epitopes of PDCoV are currently unidentified, and no information about T cell epitopes is available. Here, T-cell epitopes of PDCoV structural proteins were predicted using computational methods. 17 epitope peptides were synthesized and then screened using ELIspot, intracellular cytokine staining (ICS), and RT-qPCR detection of IFN-γ mRNA to evaluate their ability to elicit interferon-gamma (IFN-γ) responses in peripheral blood mononuclear cells (PBMCs) from PDCoV-challenged pigs. Five peptides (M1, M2, M3, N6, and S4) elicited high levels of IFN-γ and were investigated further as potential T-cell epitope candidates. All five peptides were cytotoxic T lymphocyte (CTL) epitopes, and two peptides (M3, N6) were recognized simultaneously by CD8 + and CD4 + T cells. A multi-epitope peptide combining the five epitopes (designated "5T") was synthesized and its immune response and protection efficacy was evaluated in a piglet model. ELISpot assay results indicated that 5T induces robust epitope-specific cellular immune responses. Four epitopes (M1, M2, N6, S4) elicited IFN-γ responses in 5T-vaccinated piglets. No obvious protection efficacy was detected in piglets vaccinated with 5T alone. Our results provide valuable information concerning PDCoV-related antigenic epitopes and will be useful in the design of epitope-based vaccines.

Gut microbiota modulate distal symmetric polyneuropathy in patients with diabetes.

The pathogenic mechanisms underlying distal symmetric polyneuropathy (DSPN), a common neuropathy in patients with diabetes mellitus (DM), are not fully understood. Here, we discover that the gut microbiota from patients with DSPN can induce a phenotype exhibiting more severe peripheral neuropathy in db/db mice. In a randomized, double-blind, and placebo-controlled trial (ChiCTR1800017257), compared to 10 patients who received placebo, DSPN was significantly alleviated in the 22 patients who received fecal microbiota transplants from healthy donors, independent of glycemic control. The gut bacterial genomes that correlated with the Toronto Clinical Scoring System (TCSS) score were organized in two competing guilds. Increased guild 1, which had higher capacity in butyrate production, and decreased guild 2, which harbored more genes in synthetic pathway of endotoxin, were associated with improved gut barrier integrity and decreased proinflammatory cytokine levels. Moreover, matched enterotype between transplants and recipients showed better therapeutic efficacy with more enriched guild 1 and suppressed guild 2. Thus, changes in these two competing guilds may play a causative role in DSPN and have the potential for therapeutic targeting.

Identification of an immunodominant neutralizing epitope of porcine Deltacoronavirus spike protein.

Porcine deltacoronavirus (PDCoV) is a novel swine enteropathogenic coronavirus that, because of its broad host range, poses a potential threat to public health. Here, to identify the neutralizing B-cell epitopes within the S1-CTD protein, we generated three anti-PDCoV monoclonal antibodies (mAbs). Of these, the antibody designated 4E-3 effectively neutralized PDCoV with an IC50 of 3.155 µg/mL. mAb 4E-3 and one other, mAb 2A-12, recognized different linear B-cell epitopes. The minimal fragment recognized by mAb 4E-3 was mapped to 280FYSDPKSAV288 and designated S280-288, the minimal fragment recognized by mAb 2A-12 was mapped to 506TENNRFTT513, and designated S506-513. Subsequently, alanine (A)-scanning mutagenesis indicated that Asp283, Lys285, and Val288 were the critical residues recognized by mAb 4E-3. The S280-288 epitope induces PDCoV specific neutralizing antibodies in mice, demonstrating that it is a neutralizing epitope. Of note, the S280-288 coupled to Keyhole Limpet Hemocyanin (KLH) produces PDCoV neutralizing antibodies in vitro and in vivo, in challenged piglets it potentiates interferon-γ responses and provides partial protection against disease. This is the first report about the PDCoV S protein neutralizing epitope, which will contribute to research of PDCoV-related pathogenic mechanism, vaccine design and antiviral drug development.

Effects of comfort nursing on pain and quality of life in children undergoing tonsillectomy.

OBJECTIVE: To observe the effect of comfort nursing on pain, quality of life, and nutritional status in children undergoing tonsillectomy. METHODS: In this retrospective study, a total of 114 children who underwent tonsillectomy in Chun'an Hospital of Traditional Chinese Medicine were divided into a research group and a control group according to the nursing methods, with 57 cases in each group. The control group received routine nursing care, and the research group received additional comfort nursing. We compared the pain level (assessed by visual analogue scale (VAS) scale), quality of life (assessed by Generic Quality of Life Inventory-74 (GQOLI-74)), relevant clinical indicators (postoperative swallowing recovery time, wake-up time, and hospital stay), nutrition indicators (total blood protein and albumin), sleep quality (assessed by Pittsburgh Sleep Quality Index (PAQI)), nursing satisfaction, and postoperative complications between the two groups. RESULTS: After postoperative nursing, the VAS scores and PSQI scores were significantly decreased (both P<0.05), and the GQOLI-74 scores were significantly increased (P<0.05) in both groups. The postoperative swallowing recovery time, wake-up time, and hospital stay in the research group were significantly shorter than those in the control group (all P<0.05). The levels of total serum protein and albumin in the research group were significantly higher than those in the control group (both P<0.05). The research group showed a significantly higher satisfaction rate and lower incidence of complications as compared with the control group (both P<0.05). The results of the logistic regression analysis showed that postoperative upper respiratory infection and the degree of tonsillar embedment were independent risk factors for hemorrhage after tonsillectomy (P<0.05). CONCLUSION: In children undergoing tonsillectomy, providing comfort nursing can significantly reduce pain and improve their quality of life.

Arthroscopic-assisted reduction plus internal fixation and traditional open reduction plus internal fixation for talus fractures: a comparative study.

OBJECTIVE: To explore the efficacy of arthroscopic-assisted reduction and internal fixation (ARIF) and traditional open reduction and internal fixation in the treatment of talus fractures. METHODS: This study retrospectively analyzed the clinical data of 92 patients with talus fractures admitted to our hospital. The patients were divided into a control group (treated with traditional open reduction and internal fixation) and a research group (with ARIF) with 46 cases in each. The operation indices, the score of the American Orthopaedic Foot and Ankle Society Ankle-Hindfoot Scoring System (AOFAS-AH), callus growth score, pain score, treatment effect, complications and quality of life score were compared between the two groups. RESULTS: The research group showed shorter time of fracture healing, hospitalization and less intraoperative blood loss than the control group (all P<0.001). The ankle-hindfoot score in the research group was higher than those in the control group 3 and 6 months after surgery (both P<0.001). The excellent and good rate of treatment in the research group (93.48%) was higher than that in the control group (78.26%; P<0.05). Compared with the control group, the VAS score was lower and the callus growth score was higher in the research group at 1st, 3rd and 6th month after surgery (all P<0.01). The incidence of complications in the research group (2.17%) was lower than that in the control group (13.04%; P<0.05). Six months after surgery, the SF-36 score increased compared with that before surgery, with higher parameters in the research group than in the control group (P<0.001). CONCLUSION: ARIF is more effective than traditional open reduction and internal fixation in treating talus fractures, with less complications and higher safety.

Cardiovascular Benefits of Empagliflozin Are Associated With Gut Microbiota and Plasma Metabolites in Type 2 Diabetes.

CONTEXT: Cardiovascular benefits of empagliflozin in patients with type 2 diabetes mellitus (T2DM) have been reported; however, the underlying mechanism remains unknown. OBJECTIVE: We hypothesized that the cardiovascular benefits of empagliflozin are associated with altered gut microbiota and plasma metabolites, and that empagliflozin may be used as an initial treatment for patients with T2DM at risk of cardiovascular diseases (CVDs). METHODS: This randomized, open-label, 3-month, 2-arm clinical trial included 76 treatment-naïve patients with T2DM and risk factors for CVD who were treated with either empagliflozin (10 mg/d, n = 40) or metformin (1700 mg/d, n = 36). We investigated changes in clinical parameters related to glucose metabolism and CVD risk factors, gut microbiota using 16S rRNA gene sequencing, and plasma metabolites using LC-MS. RESULTS: We found significant and similar reduction in HbA1c levels and alleviation of glucose metabolism in both groups. However, only empagliflozin improved CVD risk factors. Empagliflozin significantly reshaped the gut microbiota after 1 month of treatment; this alteration was maintained until the end of the trial. Empagliflozin increased the levels of plasma metabolites such as sphingomyelin, but reduced glycochenodeoxycholate, cis-aconitate, and uric acid levels. Concurrently, empagliflozin elevated levels of short-chain fatty acid-producing bacteria such as species from Roseburia, Eubacterium, and Faecalibacterium, and reduced those of several harmful bacteria including Escherichia-Shigella, Bilophila, and Hungatella. CONCLUSION: Empagliflozin may be a superior initial therapy for patients with T2DM at risk of CVDs; its cardiovascular benefits may be associated with shifts in gut microbiota and plasma metabolites.

Aberrant expression of inhibitory receptors on B cells in patients with Graves' disease.

The pathophysiological mechanism underlying Graves' disease (GD) remains incompletely understood. Inhibitory receptors on B cells are critical for humoral immunity, which plays a key role in GD pathogenesis. This study aimed to investigate B cell subsets distribution and inhibitory receptor expression on these subsets in GD patients. Peripheral blood was drawn from 41 healthy controls and 46 GD patients (21 patients with moderate GD, 25 patients with severe GD). B cell subset distribution and CD22, CD32b and CD72 expression on B cells were analyzed by flow cytometry. Serum cytokines were examined by enzyme-linked immunosorbent assay (ELISA). Compared with healthy controls, the naïve B cell percentage was increased, while the preswitched memory and conventional memory B cell percentages were decreased. The inhibitory receptors expression, especially CD32b, on B cell subsets was significantly decreased in patients with GD. In addition, the inhibitory receptors expression on B cell subsets from severe GD patients exhibited a decreasing trend compared with those from moderate GD patients. These results suggest that abnormal B cell subset distribution occurs in GD. Impaired inhibitory receptors, in particular CD32b, play a crucial role in GD pathogenesis and might be a therapeutic target to rebuild self-immune tolerance in GD.

Characteristics of the Gut Microbiota and Metabolism in Patients With Latent Autoimmune Diabetes in Adults: A Case-Control Study.

OBJECTIVE: Type 1 and type 2 diabetes are associated with gut dysbiosis. However, the relationship between the gut microbiota and latent autoimmune diabetes in adults (LADA), sharing clinical and metabolic features with classic type 1 and type 2 diabetes, remains unclear. Here, we used a multiomics approach to identify the characteristics of the gut microbiota and metabolic profiles in patients with LADA. RESEARCH DESIGN AND METHODS: This age- and sex-matched case-control study included 30 patients with LADA, 31 patients with classic type 1 diabetes, 30 patients with type 2 diabetes, and 29 healthy individuals. The gut microbiota profiles were identified through the 16S rRNA gene, and fecal and serum metabolites were measured through untargeted liquid chromatography-mass spectrometry. RESULTS: Patients with LADA had a significantly different structure and composition of the gut microbiota and their metabolites as well as a severe deficiency of short-chain fatty acid-producing bacteria. The gut microbiota structure of the patients with LADA was more similar to that of patients with type 1 diabetes who were positive for GAD antibody. We identified seven serum metabolite modules and eight fecal metabolite modules that differed between the LADA group and the other groups. CONCLUSIONS: The characteristic gut microbiota and related metabolites of patients with LADA are associated with autoantibodies, glucose metabolism, islet function, and inflammatory factors, which may contribute to the pathogenesis of LADA. Future longitudinal studies should explore whether modulating the gut microbiota and related metabolites can alter the natural course of autoimmune diabetes in the quest for new therapeutics.

Investigation on the micromorphology and thermophysical properties of NaNO3 heat storage materials modified by solution combustion.

Nitrate has a wide temperature range, wide operating temperature, low vapor pressure, low cost, strong heat transfer and stable chemical properties. It is widely used in solar thermal power generation heat storage material. In this paper, the alkali salt NaNO3 was modified by solution combustion method with citric acid as fuel. The structure and thermal properties of the prepared salts were studied by field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC). The results show that the solution combustion process improves the structure and thermal properties of NaNO3, and the resulting product has a new phase. The particle size and microscopic morphology of the prepared salt were changed. As the proportion of fuel increases, the hollow cuboid structure gradually grows on the surface and inside of the modified salt. The microstructure obtained is different at different ignition temperatures, and a finer and even rod-like structure is obtained at an ignition temperature of 600 °C. The specific heat capacity of all modified samples has been improved, among which solid specific heat and liquid specific heat have increased the most, respectively 3.10 J/g·K and 3.19 J/g·K, which are 140.31% and 131.16% higher than the base salt, respectively. This work not only studies the specific heat capacity of NaNO3 modified by solution combustion, but also explores the effect of micromorphology and new phase formation on its performance, which provides innovative ideas for improving the specific heat capacity of molten salt heat storage materials.

Association of serum CTRP9 levels with cardiac autonomic neuropathy in patients with type 2 diabetes mellitus.

AIMS: Cardiac autonomic neuropathy (CAN) is a serious complication of diabetes and is associated with adipokines. The C1q tumor necrosis factor-related protein 9 (CTRP9) is a newly discovered adipokine. This study aimed to evaluate the association of serum CTRP9 levels with the prevalence and severity of CAN in patients with type 2 diabetes mellitus. MATERIALS AND METHODS: We enrolled 262 patients (aged ≥18 years) with type 2 diabetes mellitus into this study. Standard cardiovascular autonomic reflex tests (CARTs) were used to assess CAN and patients were divided into three groups accordingly: a non-CAN group, an early CAN group, and a definite CAN group. Serum CTRP9 levels were measured by enzyme-linked immunosorbent assay, and the tertiles were calculated. RESULTS: Serum CTRP9 levels decreased significantly in the early CAN and definite CAN groups (P < 0.05). The percentage of definite CAN was the highest at the minimum tertile of serum CTRP9 level (T1; P < 0.05). Additionally, serum CTRP9 levels were negatively correlated with age, DM duration, hemoglobin A1c (HbA1c), and fasting plasma glucose (FPG) while positively correlated with high-density lipoprotein cholesterol (HDL; P < 0.05). The level of CTRP9 was also significantly associated with the four indexes of CARTs (P < 0.05). Furthermore, CTRP9 was a protective factor for definite CAN (P < 0.05). Compared with the maximum tertile (T3) of the serum CTRP9 levels, a decreased level of serum CTRP9 in T1 significantly increased the prevalence ratio of definite CAN in patients with type 2 diabetes mellitus (P < 0.05). CONCLUSION: Serum CTRP9 levels were independently associated with definite CAN. CTRP9 represents a reliable biomarker for exploring CAN in patients with type 2 diabetes mellitus.

Association of time in range, as assessed by continuous glucose monitoring, with painful diabetic polyneuropathy.

AIMS/INTRODUCTION: This study aimed to evaluate the association between time in range (TIR) obtained from continuous glucose monitoring and the prevalence and degree of painful diabetic neuropathy. MATERIALS AND METHODS: A total of 364 individuals with diabetic peripheral neuropathy were enrolled in this study. Sensor-based flash glucose monitoring systems were used to monitor the participants' glucose levels, and the glycemic variability metrics were calculated, including the TIR, glucose coefficient of variation, standard deviation and the mean amplitude of glycemic excursions. The participants were asked to record any form of pain during the 2 weeks of monitoring, and score the pain every day on a numerical rating scale. Based on the numerical rating scale, the patients were divided into the pain-free group, mild pain group and moderate/severe pain group. RESULTS: Overall, 51.92% (189/364) of the participants were diagnosed with painful diabetic neuropathy. Compared with the pain-free group, the level of TIR decreased significantly in the mild pain and moderate/severe pain groups (P < 0.05). The prevalence of mild pain and moderate/severe pain decreased with increasing TIR quartiles (all P < 0.05). Multiple linear regression analysis showed that TIR was significantly negatively correlated with the numerical rating scale score after adjustment for glycated hemoglobin, glycemic variability indicators and other risk factors (P < 0.05). Logistic regression analysis showed that a decreasing level of TIR was significantly associated with an increasing risk of any pain and moderate/severe pain (P < 0.05). CONCLUSIONS: TIR is correlated with painful diabetic neuropathy and is underscored as a valuable clinical evaluation measure.

Alcanivorax sediminis sp. nov., isolated from deep-sea sediment of the Pacific Ocean.

A taxonomic study was carried out on strain PA15-N-34T, which was isolated from deep-sea sediment of Pacific Ocean. The bacterium was Gram-stain-positive, oxidase- and catalase-positive and rod-shaped. Growth was observed at salinity of 0-15.0% NaCl and at temperatures of 10-45 °C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain PA15-N-34T belonged to the genus Alcanivorax, with the highest sequence similarity to Alcanivorax profundi MTEO17T (97.7 %), followed by Alcanivorax nanhaiticus 19 m-6T (97.3 %) and 12 other species of the genus Alcanivorax (93.4 %-97.0 %). The average nucleotide identity and DNA-DNA hybridization values between strain PA15-N-34T and type strains of the genus Alcanivorax were 71.46-81.78% and 18.7-25.2 %, respectively. The principal fatty acids (>10 %) were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c; 31.2 %), C16 : 0 (25.0 %) and summed feature 3 (14.6 %). The DNA G+C content was 57.15 mol%. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, four unidentified aminolipids and three unidentified lipids. The novel strain can be differentiated from its closest type strain by a negative test for urease and the presence of diphosphatidylglycerol and aminolipid. The combined genotypic and phenotypic data show that strain PA15-N-34T represents a novel species within the genus Alcanivorax, for which the name Alcanivorax sediminis sp. nov. is proposed, with the type strain PA15-N-34T (=MCCC 1A14738T=KCTC 72163T).

Association of masked uncontrolled hypertension and cardiovascular diseases in treated hypertensive patients.

INTRODUCTION: The aim of the study was to evaluate the association of masked uncontrolled hypertension (MUCH) and prevalence of cardiovascular disease in treated hypertensive patients. MATERIAL AND METHODS: Patients' demographics and prior medical histories were collected. Fasting venous blood was drawn for evaluation of serum creatinine level, which was used to calculate glomerular filtration rate (GFR). Clinic blood pressure (BP) and 24 h ambulatory blood pressure monitoring (ABPM) measurements were performed. Based on the clinic BP and 24 h ABPM results, patients were divided into MUCH and non-masked hypertension groups. RESULTS: Compared to patients without masked hypertension, MUCH patients were older (62.4 ±11.2 vs. 59.7 ±10.4 years, p < 0.05), more likely to be male (66.9% vs. 63.4%), had diabetes (33.9% vs. 29.6%), longer hypertension duration (12.4 ±5.3 vs. 9.5 ±4.5 years, p < 0.05), lower GFR (79.5 ±11.6 vs. 82.4 ±10.3 ml/min/1.73 m2, p < 0.05), treated with ß-blocker (39.0% vs. 32.7%, p < 0.05) and required more antihypertensive medications (2.7 ±0.5 vs. 2.2 ±0.3, p < 0.05). MUCH patients have higher cardiovascular disease prevalence than that without masked hypertension (30.1% vs. 23.4%, p < 0.05). After adjustment for covariates, MUCH was still independently associated with higher cardiovascular disease prevalence with odds ratio 1.38 (95% confidence interval 1.17-1.62, p < 0.05). CONCLUSIONS: The MUCH is independently associated with prevalent cardiovascular disease in treated hypertensive patients. Future studies are needed to evaluate whether correction of MUCH can improve patients' outcomes.

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The application of microbial fertilizer plays an important role in improving soil restoration and fertilizer utilization. The effects of microbial fertilizer are greatly affected by crop genotypes and ecological conditions. Little is known about the effects of microbial fertilizers on maize production in Northeast China. To develop microbial fertilizer specific to the black soil and the climate characteristics of Northeast China, we isolated five plant rhizosphere-promoting bacteria (PGPR), named as MZ1, MZ2, MZ3, MZ4 and MZ5, with different degrees of biological functions such as IAA synthesis, phosphate-solubilizing, potassium-solubilizing and siderophore-releasing, from the rhizosphere of maize field. The analysis of ecological adaptability showed that those five strains differed in salt resistance, drought tolerance, acid and alkali resistance, pesticide resistance. The 16S rRNA gene sequences analysis showed that the strains MZ1, MZ2, MZ3, MZ4 and MZ5 belonged to the genus of Sphingomonas, Enterobacter, Pseudomonas, Bacillus and Rhizobium, respectively. In maize field experiment with 50% nitrogen fertilizer reduction, the inoculation with MZ1, MZ3 and MZ5 increased grain yield by 19.9%-25.0%. MZ1, MZ3, and MZ5 could be used as microbial fertilizers for maize in Northeast China.

[Analysis of CYP17A1 gene variants in 5 patients with 17-hydroxylase deficiency].

OBJECTIVE: To identify pathogenic variants in 5 sporadic patients and two Chinese pedigrees affected with 17-hydroxylase deficiency (17-OHD). METHODS: Peripheral blood samples were collected with informed consent. Variants of CYP17A1 gene were screened by PCR and Sanger sequencing. Suspected mutations were validated in other members of the pedigrees. RESULTS: Gene sequencing has identified a homozygous c.985_987delTACinsAA (Y329Kfs) mutation in exon 6 of the CYP17A1 gene in 4 patients and the sister of case 3. Case 1 was found to harbor compound heterozygous mutations c.1459_1467del9 (p.D487_F489del) and c.1244-3C>A. The parents and brother of cases 2 and 5 were heterozygous carriers of a c.985_987delTACinsAA(Y329Kfs) mutation. CONCLUSION: Mutations of the CYP17A1 gene probably underlie the pathogenesis of 17-OHD, for which c.985_987delTACinsAA(Y329Kfs) is the most common. The c.1244-3C>A is a novel mutation. Above results have facilitated genetic counseling for the affected families.

Chengkuizengella marina sp. nov., isolated from deep-sea sediment of the Pacific Ocean.

A taxonomic study was carried out on strain YPA3-1-1T, which was isolated from deep-sea sediment of the Pacific Ocean. The bacterium was Gram-stain-positive, oxidase-positive, catalase-negative, rod-shaped and spore-forming. Growth was observed at salinities of 1.0-6.0 % and at temperatures of 10-40 °C. The isolate could degrade gelatin and aesculin. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YPA3-1-1T belonged to the genus Chengkuizengella, with the highest sequence similarity to the only typespecies, Chengkuizengella sediminis J15A17T (98.5 %). The estimated average nucleotide identity and DNA-DNA hybridization values between strain YPA3-1-1T and C. sediminis J15A17T were 88.1 and 35.0 %, respectively. The cell wall of strain YPA3-1-1T contained meso-diaminopimelic acid. The principal fatty acids (>10 %) were iso-C16 : 0 (35.5 %) and anteiso-C15 : 0 (17.5 %). The G+C content of the chromosomal DNA was 33.1 mol%. The respiratory quinone was determined to be MK-7 (100 %). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, glycolipid and three unidentified phospholipids. The combined genotypic and phenotypic data show that strain YPA3-1-1T represents a novel species within the genus Chengkuizengella, for which the name Chengkuizengella marina sp. nov. is proposed, with the type strain YPA3-1-1T (=MCCC 1A14042T=KCTC 43019T).

Integrated analysis of transcriptome data revealed MMP3 and MMP13 as critical genes in anaplastic thyroid cancer progression.

To better understand the molecular mechanisms of anaplastic thyroid carcinoma (ATC), we aimed to identify the hub genes specifically involved in ATC by integrated bioinformatics analysis. In this study, using three Gene Expression Omnibus data sets with the same platform GPL570, we screened hub genes involved in ATC progression. In vitro experiments, such as western blot analysis, Transwell assays, and coimmunoprecipitation, was performed to verify our findings. By comparing three subtypes of thyroid cancer with normal tissue, we found ATC harbored more changed genes than well and poorly differentiated thyroid cancer. Using specifically differentially expressed genes between ATC and normal thyroid tissues to perform Gene ontology (GO) analysis, ATC showed enrichments of GO terms involved in lymphocyte migration and activation, collagen catabolic and metabolic process, thyroid hormone synthesis, and embolism. Using genes involved in extracellular matrix, coexpression network analysis and protein-protein interaction analysis were performed to identify matrix metalloproteinase 3 (MMP3) and MMP13 as two hub genes. Our experimental data indicated that both MMP3 and MMP13 were upregulated in ATC and knockdown of either of them could notably suppress ATC cell invasion and migration. Mechanistically, Gene Set Enrichment Analysis, coimmunoprecipitation, and rescue experiments revealed MMP3 and MMP13 not only interacted with each other, but also regulated each other through the janus kinase/signal transducer and activator of transcription 3 and mammalian target of rapamycin pathways. In conclusion, we identified a specific molecular mechanisms for the development of ATC by integrated analysis of transcriptome and in vitro experiments, which suggested that MMP3 and MMP13 might be developed as novel therapeutic targets for ATC.