The endoplasmic reticulum membrane protein complex subunit Emc6 is essential for rhodopsin localization and photoreceptor cell survival.

The endoplasmic reticulum (ER) membrane protein complex (EMC) is responsible for monitoring the biogenesis and synthetic quality of membrane proteins with tail-anchored or multiple transmembrane domains. The EMC subunit EMC6 is one of the core members of EMC and forms an enclosed hydrophilic vestibule in cooperation with EMC3. Despite studies demonstrating that deletion of EMC3 led to rhodopsin mislocalization in rod photoreceptors of mice, the precise mechanism leading to the failure of rhodopsin trafficking remains unclear. Here, we generated the first rod photoreceptor-specific knockout of Emc6 (RKO) and cone photoreceptor-specific knockout of Emc6 (CKO) mouse models. Deficiency of Emc6 in rod photoreceptors led to progressive shortening of outer segments (OS), impaired visual function, mislocalization and reduced expression of rhodopsin, and increased gliosis in rod photoreceptors. In addition, CKO mice displayed the progressive death of cone photoreceptors and abnormal localization of cone opsin protein. Subsequently, proteomics analysis of the RKO mouse retina illustrated that several cilium-related proteins, particularly anoctamin-2 (ANO2) and transmembrane protein 67 (TMEM67), were significantly down-regulated prior to OS degeneration. Detrimental rod photoreceptor cilia and mislocalized membrane disc proteins were evident in RKO mice. Our data revealed that in addition to monitoring the synthesis of rhodopsin-dominated membrane disc proteins, EMC6 also impacted rod photoreceptors' ciliogenesis by regulating the synthesis of membrane proteins associated with cilia, contributing to the mislocalization of membrane disc proteins.

Bioinformatics analysis of CUL2/4A/9 and its function in head and neck squamous cell carcinoma.

INTRODUCTION: Several previous studies have shown that differential expression of cullin (CUL) family proteins may be involved in mediation of the signal transduction pathways associated with cancer. However, the function of CULs is still unclear in head and neck squamous cell carcinoma (HNSCC). MATERIAL AND METHODS: We used The Cancer Genome Atlas (TCGA) database, cBioPortal, Metascape, STRING, Cytoscape, Tumor Immune Estimation Resource (TIMER), Kaplan-Meier plotter, and Tumor Immune System Interaction Database (TISIDB) to access the expression of CULs and the possible correlation with the tumourigenesis, development, prognosis, immunity, and transcriptional level of CULs in HNSCC. Furthermore, real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect messenger ribonucleid acid (mRNA) levels in HNSCC tissues and cell samples. We also explored the cell proliferation and migration separately by CCK8 assay and wound healing assay. RESULTS: The results showed that the expressions of CUL2/4A were upregulated and CUL9 was downregulated in HNSCC patients as compared with normal patients. CUL2/4A/9 were also linked to the clinicopathological features and overall survival of HNSCC in bioinformatics analysis. Moreover, we noticed that CUL2/4A/9 may take part in tumour-specific immune response by modulating the tumour-infiltrating lymphocytes (TILs) and immunomodulators. Lastly, we found that CUL2/4A/9 could promote cellular proliferation and migration. CONCLUSION: These results suggest that the transcriptional levels of CUL2/4A/9 were upregulated and these genes could affect proliferation and migration of HNSCC cells. Therefore, CUL2/4A/9 could potentially function as novel independent biomarkers in HNSCC patients.

Changes in the gut microbial profile during long-term androgen deprivation therapy for prostate cancer.

BACKGROUND: Recent studies have highlighted the association between androgen deprivation therapy (ADT) and the gut microbiota in prostate cancer. However, the impact of long-term ADT remains to be explored. METHODS: To examine changes in the gut microbial profile from short-term (a median of 7 months), and middle-term (a median of 18 months) to long-term ADT (>33 months), 16S rRNA data from 56 fecal samples were reanalyzed. Additionally, a two-sample Mendelian randomization was employed to investigate the relationships between particular microbial signatures and prostate cancer as well as testosterone levels. RESULTS: In contrast to the short- and middle-term ADT groups, the long-term ADT group had significant changes in alpha and beta diversity. In particular, the relative abundance of genera such as Catenibacterium and Holdemanella decreased in the long-term ADT group, whereas the opportunistic bacterium (Erysipelatoclostridium) and Ruminococcus gnavus showed increased abundance over ADT time. Moreover, a two-sample Mendelian randomization analysis revealed the negative associations between genetically predicated genera Coprobacter, Ruminococcaceae UCG002/011, and Defluviitaleacea-UCG-011, and testosterone levels. CONCLUSIONS: In conclusion, long-term ADT use in prostate cancer patients was associated with detrimental changes in gut microbiota, including an increase in genera related to testosterone synthesis and opportunistic bacteria. These changes may be related to disease progression and side effects of long-term ADT while further longitudinal studies are required to prove this relationship.

The added effects of cold spells on stroke admissions: Differential effects on ischemic and hemorrhagic stroke.

BACKGROUND: Epidemiological evidence suggests an association between low ambient temperature and stroke risk, but available data is limited particularly on associations with different stroke subtypes. AIMS: To estimate the relationship between cold spells and stroke admissions, including the effect of cold spells on different stroke subtypes (ischaemic stroke and intracerebral haemorrhage (ICH)). METHODS: 144,405 stroke admissions from the Tianjin Centre for Health and Meteorology Multidisciplinary Innovation in China, covering the period from January 2016 to December 2020, were studied, as well as meteorological and air pollutant data. . A generalized additive model with a distributed lag nonlinear model was employed to assess the relationship, considering 12 different definitions of a cold-spell based on various temperature thresholds and durations. The analysis controlled for lagged and nonlinear effects of temperature. Analyses were performed on all strokes as well as ischaemic stroke and ICH). RESULTS: There was a significant increase in stroke admissions during cold spells. Generally, the increased risk during cold spells increased as the temperature threshold decreased, but was not significantly affected by the duration. The optimal model was obtained using the cold-spell definition based on an average daily temperature below the 10th percentile (0.11℃) for 2 or more consecutive days. According to this model, the effect of cold spells on ischemic stroke admissions had a significant lag effect and was long-lasting, with a single-day effect occurring on lag 7d, peaking on lag 13d (RR=1.05 95%CI:1.02-1.09), and lasting until lag 20d. In contrast, the effect on ICH was immediate and short-lived, with the most significant single-day effect occurring on the current day (RR=1.17 95%CI:1.06-1.29) and limited within three days. 14.15% of stroke cases could be attributed to cold spells, with ICH exhibiting a higher burden than ischemic stroke except for strict temperature threshold definitions. CONCLUSIONS: Cold spells are associated with an increased stroke risk. Different patterns of association were seen for different stroke subtypes. The effect on ischemic stroke had a lag effect and a longer duration, whereas the effect on ICH had an immediate effect and a shorter duration. These findings support the development and improvement of stroke cold-spell early warning systems and highlight the importance of public health interventions to mitigate the adverse health impacts of cold spells.

Integrated clustering signature of genomic heterogeneity, stemness and tumor microenvironment predicts glioma prognosis and immunotherapy response.

BACKGROUND: Glioma is the most frequent primary tumor of the central nervous system. The high heterogeneity of glioma tumors enables them to adapt to challenging environments, leading to resistance to treatment. Therefore, to detect the driving factors and improve the prognosis of glioma, it is essential to have a comprehensive understanding of the genomic heterogeneity, stemness, and immune microenvironment of glioma. METHODS: We classified gliomas into various subtypes based on stemness, genomic heterogeneity, and immune microenvironment consensus clustering analysis. We identified risk hub genes linked to heterogeneous characteristics using WGCNA, LASSO, and multivariate Cox regression analysis and utilized them to create an effective risk model. RESULTS: We thoroughly investigated the genomic heterogeneity, stemness, and immune microenvironment of glioma and identified the risk hub genes RAB42, SH2D4A, and GDF15 based on the TCGA dataset. We developed a risk model utilizing these genes that can reliably predict the prognosis of glioma patients. The risk signature showed a positive correlation with T cell exhaustion and increased infiltration of immunosuppressive cells, and a negative correlation with the response to immunotherapy. Moreover, we discovered that SH2D4A, one of the risk hub genes, could stimulate the migration and proliferation of glioma cells. CONCLUSIONS: This study identified risk hub genes and established a risk model by analyzing the genomic heterogeneity, stemness, and immune microenvironment of glioma. Our findings will facilitate the diagnosis and prediction of glioma prognosis and may lead to potential treatment strategies for glioma.

Single-cell RNA-sequencing reveals heterogeneity and intercellular crosstalk in human tuberculosis lung.

Lung inflammation indicated by 18F-labeled fluorodeoxyglucose (FDG) in patients with tuberculosis is associated with disease severity and relapse risk upon treatment completion. We revealed the heterogeneity and intercellular crosstalk in lung tissues with 18F-FDG avidity and adjacent uninvolved tissues from 6 tuberculosis patients by single-cell RNA-sequencing. Tuberculous lungs had an influx of regulatory T cells (Treg), exhausted CD8 T cells, immunosuppressive myeloid cells, conventional DC, plasmacytoid DC, and neutrophils. Immune cells in inflamed lungs showed general up-regulation of ATP synthesis and interferon-mediated signaling. Immunosuppressive myeloid and Treg cells strongly displayed transcriptions of genes related to tuberculosis disease progression. Intensive crosstalk between IL4I1-expressing myeloid cells and Treg cells involving chemokines, costimulatory molecules, and immune checkpoints, some of which being specific in 18F-FDG-avid lungs, were found. Our analysis provides insights into the transcriptomic heterogeneity and cellular crosstalk in pulmonary tuberculosis and guide unveiling cellular and molecular targets for tuberculosis therapy.

A new time-varying coefficient regression approach for analyzing infectious disease data.

Since the beginning of the global pandemic of Coronavirus (SARS-COV-2), there has been many studies devoted to predicting the COVID-19 related deaths/hospitalizations. The aim of our work is to (1) explore the lagged dependence between the time series of case counts and the time series of death counts; and (2) utilize such a relationship for prediction. The proposed approach can also be applied to other infectious diseases or wherever dynamics in lagged dependence are of primary interest. Different from the previous studies, we focus on time-varying coefficient models to account for the evolution of the coronavirus. Using two different types of time-varying coefficient models, local polynomial regression models and piecewise linear regression models, we analyze the province-level data in Canada as well as country-level data using cumulative counts. We use out-of-sample prediction to evaluate the model performance. Based on our data analyses, both time-varying coefficient modeling strategies work well. Local polynomial regression models generally work better than piecewise linear regression models, especially when the pattern of the relationship between the two time series of counts gets more complicated (e.g., more segments are needed to portray the pattern). Our proposed methods can be easily and quickly implemented via existing R packages.

In Situ N, O-Dually Doped Nanoporous Biochar Derived from Waste Eutrophic Spirulina for High-Performance Supercapacitors.

Sustainable and high-performance energy storage materials are crucial to address global energy and environmental challenges. In this study, Spirulina platensis was used as the carbon and nitrogen source, and Spirulina-based nanoporous biochar (SNPB) was synthesized through chemical activation using KOH as the activating agent in N2 atmosphere. SNPB-800-4 was characterized by N2 adsorption-desorption and XPS, showing a high specific surface area (2923.7 m2 g-1) and abundant heteroatomic oxygen (13.78%) and nitrogen (2.55%). SNPB-800-4 demonstrated an exceptional capacitance of 348 F g-1 at a current density of 1 A g-1 and a remarkable capacitance retention of 94.14% after 10,000 cycles at a current density of 10 A g-1 in 6 M KOH. Notably, symmetric supercapacitors SNPB-800-4//SNPB-800-4 achieved the maximum energy and power densities of 17.99 Wh kg-1 and 162.48 W kg-1, respectively, at a current density of 0.5 A g-1, and still maintained 2.66 Wh kg-1 when the power density was increased to 9685.08 W kg-1 at a current density of 30 A g-1. This work provides an easily scalable and straightforward way to convert waste algae biomass into in situ N, O-dually doped biochar for ultra-high-power supercapacitors.

Calculation of left ventricular ejection fraction using an 8-layer residual U-Net with deep supervision based on cardiac CT angiography images versus echocardiography: a comparative study.

Background: Accurate segmentation of the left ventricle (LV) is an important step in assessing cardiac function. Cardiac CT angiography (CCTA) has become an important means of clinical diagnosis of cardiovascular diseases (CVDs) because of its advantages of non-invasive, short examination time and low cost. In order to obtain the segmentation of LV in CCTA scans, we propose a deep learning method based on 8-layer residual U-Net with deep supervision. In this study we compared the left ventricular ejection fraction (LVEF) calculated by deep learning (DL) method (AccuLV) from CCTA to LVEF by conventional two-dimensional echocardiography (EC). Methods: This was a retrospective cross-sectional study, and consecutive patients who had undergone CCTA and EC in our hospital from February 2021 to May 2021 were recruited. The current study included 180 patients who had undergone CCTA and EC. To obtain LVEF, we used an 8-layer residual U-Net with deep supervision to segment LV contours from CCTA images and compute LVEF (DL-LVEF). The EC and DL-LVEF measurements were compared. A 50% EC-LVEF cut-off value was used as a reference standard to assess the diagnostic performance of AccuLV in assessing LV function. Results: The overall mean EC-LVEF and DL-LVEF values were 64.0% (52.3%, 69.0%) and 73.0% (52.3%, 77.0%), respectively. Three patient groups were studied: (I) hypertensive patients, (II) postmenopausal women, and (III) diabetes. EC-LVEF and DL-LVEF were found to be positively correlated for all of the included patients (r=0.82, P<0.001), with the detailed results for the three groups as follows: hypertensive patients (r=0.77, P<0.001), postmenopausal women (r=0.92, P<0.001) and diabetes (r=0.88, P<0.001). The diagnostic accuracy, sensitivity, and specificity of the DL method in predicting EC-LVEF <50% for all patients were 93.9%, 92.3%, and 94.3%, and for hypertensive patients were 95.4%, 93.8%, and 95.8%, for postmenopausal women were 87.0%, 100%, and 84.2%, for diabetes were 97.4%, 100%, and 96.6%. Conclusions: In comparison to echocardiography, which is commonly used in clinical setting, AccuLV may be a promising, fully automated tool for rapid and accurate quantification of LV function and thus for making reliable clinical decisions.

Associations of metal mixtures with thyroid function and potential interactions with iodine status: results from a cross-sectional study in MEWHC.

Few studies are available on associations between metal mixture exposures and disrupted thyroid hormone homeostasis; particularly, the role of iodine status was ignored. Here, we aimed to explore the cross-sectional relationship of blood cell metals with thyroid homeostasis and explore the potential modifying effect of iodine status. Among 328 workers from the manganese-exposed workers healthy cohort (MEWHC), we detected thyroid function parameters: thyroid stimulating hormone (TSH), total triiodothyronine (TT3), free triiodothyronine (FT3), total tetraiodothyronine (TT4), free tetraiodothyronine (FT4) as well as calculated sum activity of peripheral deiodinases (GD) and thyroid's secretory capacity (GT). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure 22 metal concentrations in blood cells. Based on the consistent results of least absolute shrinkage and selection operator (LASSO) and Bayesian kernel machine regression (BKMR) analyses, there were significant positive associations between copper and TSH (ß = 2.016), iron and FT4 (ß = 0.403), titanium and GD (ß = 0.142), nickel and GD (ß = 0.057), and negative associations between copper and FT4 (ß = - 0.226), selenium and GD (ß = - 0.332), among the participants. Interestingly, we observed an inverted-U shape relationship between magnesium and FT4. Furthermore, we found a synergistic effect between arsenic and copper on the TSH level, while antagonistic effects between nickel and copper as well as nickel and selenium on the TSH level. We observed a modified effect of iodine status on association between strontium and GD (Pinteraction = 0.026). It suggests metal mixture exposures can alter thyroid homeostasis among the occupational population, and deiodinase activity had a modified effect on association between strontium and GD. Validation of these associations and elucidation of underlying mechanisms require further researches in the future.

Cornuside improves murine autoimmune hepatitis through inhibition of inflammatory responses.

BACKGROUND: Autoimmune hepatitis (AIH) poses an important public health concern worldwide, with few therapeutic options available. Cornuside, a primary cornel iridoid glycoside present in Cornus officinalis Sieb. et Zucc., is a well-known traditional Chinese medicine that possesses anti-inflammatory, antioxidant and anti-apoptotic properties. However, the effects of cornuside on autoimmune diseases including AIH is still not defined, neither is clear on the mechanisms of cornuside in the suppression of inflammatory responses. PURPOSE: The study was aimed to investigate the therapeutic effects of cornuside on AIH using murine models. STUDY DESIGN: A murine model of AIH induced by concanavalin A (Con A) was used to examine the pharmacological activity of cornuside in suppressing the inflammatory responses in vivo. METHODS: C57BL/6J mice were intravenously with different doses of cornuside and challenged with 18 mg/kg Con A 3 h later. Network pharmacological analysis was performed to identify the potential target genes and signaling pathways by cornuside in AIH. Next serum and liver tissues were collected 12 h after Con A injection to analyze the levels of markers for hepatic injury, apoptosis, oxidative stress, immune responses, and inflammation. RESULTS: Network pharmacological analysis revealed that cornuside may modulate oxidative stress and apoptosis in AIH. Compared with the Con A group, cornuside pretreatment significantly reduced the serum levels of alanine aminotransferase and aspartate aminotransferase, improving histopathological damage and apoptosis in the livers. In addition, cornuside decreased the levels of malondialdehyde, myeloperoxidase, but increased superoxide dismutase levels, suggesting the relieving of oxidative stress. Furthermore, cornuside suppressed the activation of T and natural killer T cells, whereas the proportion of myeloid-derived suppressor cells was significantly increased. The production of proinflammatory cytokines, including interleukin (IL)-6, IL-12, IL-1ß, and tumor necrosis factor-alpha (TNF-α), was also clearly decreased. Finally, western blot analysis displayed that cornuside inhibited the phosphorylation of extracellular receptor kinase (ERK) and c-Jun N-terminal kinase (JNK). CONCLUSIONS: We demonstrated that cornuside has protective effects for Con A-induced immune-mediated hepatitis by suppressing the oxidative stress, apoptosis, and the inflammatory responses through the ERK and JNK signaling pathways, as well as by modulating the activation and recruitment of immune cells.

Biochar carbon sequestration potential rectification in soils: Synthesis effects of biochar on soil CO2, CH4 and N2O emissions.

Biochar production and its soil sequestration are promising ways to mitigate global warming. Effects of biochar on soil CO2, CH4 and N2O release have been studied extensively. In contrast, few studies have comprehensively quantified and synthesized the effect of biochar on soil greenhouse gas (GHG) emission and coupled it to the calculation of carbon sequestration potential. This study obtained the influence coefficient of biochar on soil GHG release relative to biochar carbon storage potential in soils under different environmental conditions, by literature statistics and data transformations. Our results showed that the overall average effect of biochar on soil CO2, CH4,N2O and CO2e release observed in our databases would compensate the potential of biochar soil carbon storage by -2.1 ±â€¯3.3 %, 13.1 ±â€¯9.8 %, -1.6 ±â€¯8.6 % and 5.3 ±â€¯11.4 %, respectively. By combining biochar induced soil GHG emission reduction mechanism and results from our literature statistics, some specific application environmental scenarios (such as biochar with high pyrolysis temperature of 500-600 °C, application in flooded soils, application in straw-return scenarios, etc.) were recommended, which could increase the actual carbon sequestration potential of biochar by an average of about 43.3 ±â€¯30.2 % relative the amount of carbon buried. Our findings provide a scientific basis for developing a precise application strategy towards large scale adoption of biochar as a soil amendment for climate change mitigation.

Knockdown of LIMD2 inhibits the progression of ovarian carcinoma through ERK1/2 pathway.

BACKGROUND: The incidence rate of ovarian carcinoma (OC) is the third of the female reproductive system malignant tumors, while its mortality rate ranks first among causes of female reproductive system tumor related death in the world. METHODS: In the present research, we investigated the specific role of LIMD2 through LIMD2 knockdown in OC cells. RESULTS: The results of online analysis and expression detection proved that LIMD2 was up-regulated in human OC tissues and cells. Knockdown of LIMD2 inhibited the proliferation, migration and invasion in OC cells. LIMD2 knockdown promoted the apoptosis, as well as the expression of Cleaved-Caspase3 and Bax. Importantly, knockdown of LIMD2 promotes cell autophagy. LC3-II/I ratio and Beclin1 expression increased in LIMD2 knockdown cells, while P62 expression declined in LIMD2 knockdown cells. Additionally, the phosphorylation of ERK1/2 was inhibited by the knockdown of LIMD2 in SKOV3 and OVCAR3 cells. CONCLUSION: Knockdown of LIMD2 inhibits cell proliferation, migration, invasion and autophagy, and promotes the apoptosis through the ERK1/2 signaling pathway, suggesting that LIMD2-siRNA may be an effective molecule to prevent OC progression.

Engineering pedicled vascularized bladder tissue for functional bladder defect repair.

An engineered bladder construct that mimics the structural and functional characteristics of native bladder is a promising therapeutic option for bladder substitution. We previously showed that pedicled vascularized smooth muscle tissue fabricated by grafting smooth muscle cell (SMC) sheets onto an axial capsule vascular bed had the potential for reliable bladder reconstruction. In this study, we investigated the feasibility of buccal mucosa graft (BMG) integration with the pedicled vascularized smooth muscle tissue to generate a full-layer pedicled vascularized bladder construct. BMG transplanted onto vascularized smooth muscle tissue showed good survival and developed into a pedicled vascularized bladder construct with full-layer structures, appropriate thickness, abundant vascularization, and effective barrier function. Then the full-thickness bladder defects were, respectively, reconstructed by pedicled capsule tissue (pedicled capsule group), nonpedicled vascularized bladder construct (nonpedicled construct group), and pedicled vascularized bladder construct (pedicled construct group). The picrosirius red (PSR) staining and immunohistochemistry results showed minimal fibrosis, maximal smooth muscle proportion, and high vascular density in the pedicled construct group. A continuous mucosal layer was observed only in the pedicled construct group. Moreover, morphological and functional studies revealed better bladder compliance and good ductility in the pedicled construct group. Overall, these results suggested that the BMG could be well integrated with vascularized smooth muscle tissue and generated a pedicled, fully vascularized, and structurally organized bladder construct, which facilitated structural remodeling and functional recovery and could become an alternative to bladder reconstruction.

[Source Apportionment and Source-specific Risk of Typical Antibiotics in Baiyangdian Lake].

The current situation of antibiotic pollution in lakes is critical. At present, most of the previous studies on antibiotics in lakes have focused on the spatiotemporal distribution and risk assessment, while less attention has been paid to the source apportionment. Ultra-high performance liquid chromatography-mass spectrometry was used to determine the concentration of tetracyclines (TCs), sulfonamides (SAs), and quinolones (QNs) in the samples. The source apportionment and source-specific risk of typical antibiotics in the study area were analyzed using the combination of a PMF model and risk quotients (RQ). The results showed that ① the total concentrations of target antibiotics (Σ antibiotics) ranged from ND to 2635 ng·L-1 for surface water and from ND to 259.8 ng·g-1 for sediments. ② The spatial distribution of QNs in surface water decreased from west to east, SAs decreased from middle to north and south, and TCs increased from middle to north and south. In the sediment, QNs decreased from middle to east and west, whereas SAs and TCs increased from east to west. ③ Aquaculture was the major antibiotic source, accounting for the highest proportion (33.2%), followed by sewage treatment plants (29.2%), livestock activities (18.9%), and domestic sewage (18.7%). ④ The ecological risk assessment results showed that enrofloxacin and flumequine were at a medium-high risk level. ⑤ For the spatial distribution of source-specific risk, the results showed that the aquaculture at S1 was at a high risk level, whereas the source-specific risks for other sites were at a medium-low risk level. In terms of source types, aquaculture was at a medium-high risk level, whereas the other sources were at a medium-low risk level. Therefore, considering the major sources and source-specific risk level of antibiotics, more precise and scientific antibiotic risk control should be adopted in Baiyangdian Lake.

[Source Analysis and Health Risk Assessment of Heavy Metals in the Groundwater of Shijiazhuang, a Typical City in North China Plain].

Increasing attention has been paid to the heavy metal pollution in groundwater. The source analysis and risk assessment of heavy metals will provide data and method support for the targeted control of heavy metal pollution in groundwater. In this study, 20 sampling sites were selected in Shijiazhuang City. The APCS-MLR model and health risk model were applied to analyze and evaluate the pollution sources and health risks of 10 types of heavy metals in the groundwater of Shijiazhuang. The results showed that ① the mean concentration of heavy metals in groundwater followed the order of Fe>Zn>Mn>Cu>Al>Pb>Cr>As>Cd>Hg, and the mean ρ(Fe) and ρ(Pb) were 260.3 µg·L-1 and 10.01 µg·L-1, respectively. According to the results of the single factor and Nemerow index, Pb, Fe, and Cd primarily contributed to the heavy metal pollution in the groundwater. ② The concentration of heavy metals ranged from 47.30 to 2560 µg·L-1. In terms of spatial distribution, the highest concentration appeared at S3 (2560 µg·L-1), whereas the lowest concentration was at S9 (47.30 µg·L-1). ③ Source analysis results showed that industrial and agricultural activities, transportation emission, and geological background were the major heavy metal sources, among which the contribution of industrial and agricultural activities was the highest (47.83%). ④ The industrial-agricultural activities posed a potential threat to adults (HI>1); however, the non-cancer and the cancer risks of other sources for both adults and children were at an acceptable level (HI<1) and potential threat level, respectively; industrial-agricultural activities were the major source of non-cancer (adults:52.46%, children:52.45%) and cancer risks (adults:65.22%, children:65.69%), among which Cd and As showed high cancer risk. Therefore, to ensure the safety of the groundwater environment, strictly controlling the pollution sources and further strengthening the risk control of heavy metal pollution in groundwater are necessary.

Effect of Hypoglycemic Drugs on Kisspeptin Expression in the Hypothalamic Arcuate Nucleus of PCOS Rats.

Background: To investigate the change in hypothalamic kisspeptin-1 (Kiss1) expression during the development of polycystic ovary syndrome (PCOS) and hypoglycemic drug intervention. Methods: Letrozole lavage was used to construct a polycystic ovary rat model. After successful modeling, we treated PCOS rats with metformin, pioglitazone, and acarbose, and we then observed changes in weight, estrus, glucose tolerance, insulin resistance, sex hormones, and hypothalamic kiss1 expression. Results: PCOS rats exhibited increased body weight, abnormal estrous cycle, impaired glucose tolerance, insulin resistance, increased testosterone level, increased luteinizing hormone level, and increased Kiss1 expression in the hypothalamus. However, intervention with metformin, pioglitazone, and acarbose improved the reproductive and metabolic disorders as well as reduced hypothalamic Kiss1 expression. Conclusion: The expression of hypothalamic Kiss1 may play an important role in the pathogenesis of PCOS. Metformin, pioglitazone, and acarbose may reduce the expression of hypothalamic Kiss1 by improving insulin resistance, thereby improving reproductive and metabolic disorders in PCOS rats.

Crystal structure of the GDSL family esterase EstL5 in complex with PMSF reveals a branch channel of the active site pocket.

Esterases/lipases from the GDSL family have potential applications in the hydrolysis and synthesis of important esters of pharmaceutical, food, and biotechnical interests. However, the structural and functional understanding of GDSL enzymes is still limited. Here, we report the crystal structure of the GDSL family esterase EstL5 complexed with PMSF at 2.34 Å resolution. Intriguingly, the PMSF binding site is not located at the active site pocket but is situated in a surface cavity. At the active site, we note that there is a trapped crystallization solvent 1,6-hexanediol, which mimics the bound ester chain, allowing for further definition of the active site pocket of EstL5. The most striking structural feature of EstL5 is the presence of a unique channel, which extends approximately 18.9 Å, with a bottleneck radius of 6.8 Å, connecting the active-site pocket and the surface cavity. Replacement of Ser205 with the bulk aromatic residue Trp or Phe could partially block the channel at one end and perturb its access. Reduced enzymatic activity is found in the EstL5 S205W and EstL5 S205F mutants, suggesting the functional relevance of the channel to enzyme catalysis. Our study provides valuable information regarding the properties of the GDSL-family enzymes for designing more efficient and robust biocatalysts.

Pharmacokinetics of Henagliflozin in Dialysis Patients with Diabetes.

AIM: This study aimed to assess the pharmacokinetics of henagliflozin in dialysis patients with diabetes. METHODS: In this prospective, randomized, open-label study where 10 hemodialysis and 10 peritoneal dialysis patients with diabetes were randomized in a 1:1:1:1 ratio to oral administration of henagliflozin in doses of 5 and 10 mg/day. The pharmacokinetics of a single dose of henagliflozin on Days 1 and 2, the minimum plasma concentration (Cmin) of the steady state on Day 10, and single hemodialysis clearance of henagliflozin were measured. RESULTS: The mean values of Cmax were 70.2-77.0 ng/mL and 105-143 ng/mL in the 5 mg and 10 mg henagliflozin groups, respectively; the mean values of AUCinf were 777-811 h*ng/mL and 1290-1730 h*ng/mL in the 5 mg and 10 mg henagliflozin groups, respectively. The median Tmax values ranged from 1 to 3 h across the dose range. The mean values of T1/2 of henagliflozin were 14.1-14.5 and 16.2-21.0 h in the 5 mg and 10 mg groups, respectively. The Cmin values of the steady state in dialysis patients taking 5 mg and 10 mg of henagliflozin were 15.0 ± 4.4 ng/mL and 26.8 ± 16.3 ng/mL, respectively, which were 123.8% and 131.0% higher than those in diabetic patients with normal renal function, respectively. Henagliflozin concentration was decreased by 1.1% after hemodialysis treatment. No treatment-related serious adverse events or discontinuations occurred. CONCLUSIONS: Henagliflozin at the current recommended dosage may be safe, although it is possible to result in slight accumulation in patients on dialysis. REGISTRATION: Chinese Clinical Trial Registry number ChiCTR2200062872. The date of registration: August 22, 2022.