Exploring myometrial microenvironment changes at the single-cell level from nonpregnant to term pregnant states.

The microenvironment and cell populations within the myometrium play crucial roles in maintaining uterine structural integrity and protecting the fetus during pregnancy. However, the specific changes occurring at the single-cell level in the human myometrium between nonpregnant (NP) and term pregnant (TP) states remain unexplored. In this study, we used single-cell RNA sequencing (scRNA-Seq) and spatial transcriptomics (ST) to construct a transcriptomic atlas of individual cells in the myometrium of NP and TP women. Integrated analysis of scRNA-Seq and ST data revealed spatially distinct transcriptional characteristics and examined cell-to-cell communication patterns based on ligand-receptor interactions. We identified and categorized 87,845 high-quality individual cells into 12 populations from scRNA-Seq data of 12 human myometrium tissues. Our findings demonstrated alterations in the proportions of five subpopulations of smooth muscle cells in TP. Moreover, an increase in monocytic cells, particularly M2 macrophages, was observed in TP myometrium samples, suggesting their involvement in the anti-inflammatory response. This study provides unprecedented single-cell resolution of the NP and TP myometrium, offering new insights into myometrial remodeling during pregnancy.NEW & NOTEWORTHY Using single-cell RNA sequencing and spatial transcriptomics, the myometrium was examined at the single-cell level during pregnancy. We identified spatially distinct cell populations and observed alterations in smooth muscle cells and increased M2 macrophages in term pregnant women. These findings offer unprecedented insights into myometrial remodeling and the anti-inflammatory response during pregnancy. The study advances our understanding of pregnancy-related myometrial changes.

Inversion of Molecular Chirality Associated with Ferroelectric Switching in a High-Temperature Two-Dimensional Perovskite Ferroelectric.

Controlling molecular chirality by external stimuli is of great significance in both fundamental research and technological applications. Herein, we report a high-temperature (384 K) molecular ferroelectric of a Cu(II) complex whose spontaneous polarization can be switched associated with flipping of molecular chirality. In this two-dimensional perovskite structure, the inorganic layer is separated by (NH3(CH2)2SS(CH2)2NH3)2+ organic cations skewed in a chiral conformation (P- or M-helicity in an individual crystal). As the stereodynamic disulfide bridge determines the molecular dipole moment along the polar axis, the chiral organic cation can be converted to its enantiomer as a consequence of an electric field-induced shift of the S-S moiety relative to its screw axis during the ferroelectric switching. The variation of the molecular chirality is examined with single-crystal X-ray diffraction and circular dichroism spectra. The simultaneous switching of molecular chirality and spontaneous polarization in this perovskite ferroelectric may lead to novel chiral electronic phenomena.

Dynamic viral integration patterns actively participate in the progression of BK polyomavirus-associated diseases after renal transplantation.

The classical lytic infection theory along with large T antigen-mediated oncogenesis cannot explain the BK polyomavirus (BKPyV)-associated tumor secondary to BKPyV-associated nephropathy (BKVAN), viremia/DNAemia, and viruria after renal transplantation. This study performed virome capture sequencing and pathological examination on regularly collected urine sediment and peripheral blood samples, and BKVAN and tumor biopsy tissues of 20 patients with BKPyV-associated diseases of different stages. In the early noncancerous stages, well-amplified integration sites were visualized by in situ polymerase chain reaction, simultaneously with BKPyV inclusion bodies and capsid protein expression. The integration intensity, the proportion of microhomology-mediated end-joining integration, and host PARP-1 and POLQ gene expression levels increased with disease progression. Furthermore, multiomics analysis was performed on BKPyV-associated urothelial carcinoma tissues, identifying tandem-like structures of BKPyV integration using long-read genome sequencing. The carcinogenicity of BKPyV integration was proven to disturb host gene expression and increase viral oncoprotein expression. Fallible DNA double-strand break repair pathways were significantly activated in the parenchyma of BKPyV-associated tumors. Olaparib showed an antitumor activity dose-response effect in the tumor organoids without BRCA1/2 genes mutation. In conclusion, the dynamic viral integration patterns actively participate in the progression of BKPyV-associated diseases and thus could be a potential target for disease monitoring and intervention.

Clinical features of Talaromyces marneffei infection in HIV-positive and HIV-negative individuals: A retrospective study in southern China.

Talaromyces marneffei (TSM) is a temperature-dependent dimorphic fungus endemic to Southeast Asia and southern China. As the number of people at risk of TSM infection continues to increase, the clinical manifestations are becoming increasingly complex, posing challenges for clinical management. In this study, we analyzed the medical records of 99 patients (71 human immunodeficiency virus [HIV]-positive and 28 HIV-negative) diagnosed with TSM infection from January 1, 2017, to December 31, 2022, in southern China and compared the clinical manifestations in HIV-positive and HIV-negative patients. Most patients (83/99, 84%) were male. The incidence of skin and soft tissue involvement (48% vs. 21%, P = .016); disseminated infection with blood circulation, hematopoietic, lymphatic, alimentary, or central nervous system involvement (69% vs. 36%, P = .002); and gastrointestinal bleeding (33% vs. 9%, P = .023) was higher in the HIV-positive group than the HIV-negative group. The HIV-positive group also had significantly higher alanine aminotransferase (ALT) levels (31 [26-42] vs. 14 [11-16] U/l, P < .001) and ALT/aspartate transaminase ratio (1.9 [1.5-2.2] vs. 1.3 [1.1-1.6], P = .006) than the HIV-negative group. The time to diagnosis (5.5 ± 1.1 vs. 5.1 ± 1.4 days, P = .103), antifungal regimen (P = .278), case fatality rate (20% vs. 21%, P = .849), and relapse/reinfection rate (11% vs. 19%, P = .576) did not differ significantly between the HIV-positive and HIV-negative groups. Poor antiretroviral therapy adherence (OR = 26.19, 95%CI 3.26-210.70, P = .002), advanced age (OR = 1.13, 95%CI 1.03-1.23, P = .010), and Epstein-Barr virus co-infection (OR = 37.13, 95%CI 3.03-455.64, P = .005) were independent risk factors for all-cause mortality from TSM infection in HIV-positive patients. Overall, the predominant infection sites, clinical manifestations, and complications of TSM infection differed by HIV status. However, with prompt diagnosis and appropriate treatment, HIV-positive patients with TSM infection can have similar outcomes to HIV-negative patients.

There are certain differences in the clinical features, sites of infection, and associated complications of Talaromyces marneffei infection between individuals with and without human immunodeficiency virus. It is necessary to accurately identify individuals at high risk to enable prompt diagnosis and standardized treatment.

Fatal BK polyomavirus-associated pneumonia: report of two cases with literature review.

BACKGROUND: In immunocompromised populations, such as patients with AIDS and recipients of solid organ and hematopoietic stem cell transplants, BK polyomavirus (BKPyV) can reactivate and cause several diseases, which can lead to death in their severe forms. Unlike hemorrhagic cystitis and BKPyV-associated nephropathy, BKPyV-associated pneumonia is rare, with only seven known cases worldwide. However, the disease can rapidly progress with extremely high mortality. CASE PRESENTATION: Herein, we report two cases of BKPyV-associated pneumonia following hematopoietic stem cell transplantation. Both patients had consistent infectious pneumonia and graft-versus-host disease after stem cell transplantation. The diagnosis of BKPyV-associated pneumonia was confirmed by metagenomic next-generation sequencing and polymerase chain reaction after the sudden worsening of the pulmonary infection signs and symptoms concomitant with renal dysfunction and systemic immune weakening. Both patients eventually died of systemic multi-organ failure caused by severe pneumonia. CONCLUSIONS: Currently, BKPyV reactivation cannot be effectively prevented. Immunocompromised patients must actively manage their primary lung infections, pay close attention to pulmonary signs and imaging changes. Especially during and after steroid pulse therapy or immunosuppressive therapy for graft versus host diseases, BKPyV load in blood/urine needs to be regularly measured, and the immunosuppressive intensity should be adjusted properly after the BKPyV reactivation diagnosis. Clinical trials of new antiviral drugs and therapies for BKPyV are urgently needed.

Microbiota of long-term indwelling hemodialysis catheters during renal transplantation perioperative period: a cross-sectional metagenomic microbial community analysis.

Background: Catheter-related infection (CRI) is a major complication in patients undergoing hemodialysis. The lack of high-throughput research on catheter-related microbiota makes it difficult to predict the occurrence of CRI. Thus, this study aimed to delineate the microbial structure and diversity landscape of hemodialysis catheter tips among patients during the perioperative period of kidney transplantation (KTx) and provide insights into predicting the occurrence of CRI.Methods: Forty patients at the Department of Transplantation undergoing hemodialysis catheter removal were prospectively included. Samples, including catheter tip, catheter outlet skin swab, catheter blood, peripheral blood, oropharynx swab, and midstream urine, from the separate pre- and post-KTx groups were collected and analyzed using metagenomic next-generation sequencing (mNGS). All the catheter tips and blood samples were cultured conventionally.Results: The positive detection rates for bacteria using mNGS and traditional culture were 97.09% (200/206) and 2.65% (3/113), respectively. Low antibiotic-sensitivity biofilms with colonized bacteria were detected at the catheter tip. In asymptomatic patients, no statistically significant difference was observed in the catheter tip microbial composition and diversity between the pre- and post-KTx group. The catheter tip microbial composition and diversity were associated with fasting blood glucose levels. Microorganisms at the catheter tip most likely originated from catheter outlet skin and peripheral blood.Conclusions: The long-term colonization microbiota at the catheter tip is in a relatively stable state and is not readily influenced by KTx. It does not act as the source of infection in all CRIs, but could reflect hematogenous infection to some extent.

A two-stage reconstruction method for complex networked system with hidden nodes.

Reconstructing the interacting topology from measurable data is fundamental to understanding, controlling, and predicting the collective dynamics of complex networked systems. Many methods have been proposed to address the basic inverse problem and have achieved satisfactory performance. However, a significant challenge arises when we attempt to decode the underlying structure in the presence of inaccessible nodes due to the partial loss of information. For the purpose of improving the accuracy of network reconstruction with hidden nodes, we developed a robust two-stage network reconstruction method for complex networks with hidden nodes from a small amount of observed time series data. Specifically, the proposed method takes full advantage of the natural sparsity of complex networks and the potential symmetry constraints in dynamic interactions. With robust reconstruction, we can not only locate the position of hidden nodes but also precisely recover the overall network structure on the basis of compensated nodal information. Extensive experiments are conducted to validate the effectiveness of the proposed method and superiority compared with ordinary methods. To some extent, this work sheds light on addressing the inverse problem, of which the system lacks complete exploration in the network science community.

Proteomic and miRNA Profiles of Exosomes Derived from Myometrial Tissue in Laboring Women.

Myometrial contraction is essential for successful delivery. Recent studies have highlighted the vital roles of tissue-derived exosomes in disease diagnostic, prognostic, and therapeutic applications; however, the characteristics of uterine myometrium-derived exosomes are unclear. Here, we successfully isolated exosomes from myometrial tissues, human myometrial smooth muscle cells (HMSMCs), and human umbilical vein endothelial cells (HUVECs), then performed quantitative liquid chromatography-tandem mass spectrometry and miRNA sequencing to investigate the cargo of the exosomes. Fifty-two proteins and five miRNAs were differentially expressed (DE) in term non-labor and term labor myometrium-derived exosomes. Among them, seven proteins (SERPINE1, THBS1, MGAT1, VIM, FGB, FGG, and VWF) were differentially expressed both in the myometrial exosomes and tissues, three miRNAs (miR-363-3p, miR-203a-3p, and miR-205-5p) target 13 DE genes. The top three miRNA derived from HMSMCs (miR-125b-1-3p, miR-337-5p, and miR-503-5p) and HUVECs (miR-663a, miR-4463, and miR-3622a-5p) were identified. Two proteins, GJA1 and SLC39A14, exist in female blood exosomes and are highly expressed in HMSMCs exosomes, are also upregulated in the laboring myometrium, which verified increased in laboring blood samples, might be novel potential biomarkers for myometrial activation. The proteomic and miRNA profile of exosomes derived from laboring myometrium revealed some molecules in the exosomes that affect the intercellular communication and the function of the myometrium.

Shotgun metagenomics reveals both taxonomic and tryptophan pathway differences of gut microbiota in major depressive disorder patients.

BACKGROUND: The microbiota-gut-brain axis, especially the microbial tryptophan (Trp) biosynthesis and metabolism pathway (MiTBamp), may play a critical role in the pathogenesis of major depressive disorder (MDD). However, studies on the MiTBamp in MDD are lacking. The aim of the present study was to analyze the gut microbiota composition and the MiTBamp in MDD patients. METHODS: We performed shotgun metagenomic sequencing of stool samples from 26 MDD patients and 29 healthy controls (HCs). In addition to the microbiota community and the MiTBamp analyses, we also built a classification based on the Random Forests (RF) and Boruta algorithm to identify the gut microbiota as biomarkers for MDD. RESULTS: The Bacteroidetes abundance was strongly reduced whereas that of Actinobacteria was significantly increased in the MDD patients compared with the abundance in the HCs. Most noteworthy, the MDD patients had increased levels of Bifidobacterium, which is commonly used as a probiotic. Four Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologies (KOs) (K01817, K11358, K01626, K01667) abundances in the MiTBamp were significantly lower in the MDD group. Furthermore, we found a negative correlation between the K01626 abundance and the HAMD scores in the MDD group. Finally, RF classification at the genus level can achieve an area under the receiver operating characteristic curve of 0.890. CONCLUSIONS: The present findings enabled a better understanding of the changes in gut microbiota and the related Trp pathway in MDD. Alterations of the gut microbiota may have the potential as biomarkers for distinguishing MDD patients form HCs.

Complex networks identification using Bayesian model with independent Laplace prior.

Identification of complex networks from limited and noise contaminated data is an important yet challenging task, which has attracted researchers from different disciplines recently. In this paper, the underlying feature of a complex network identification problem was analyzed and translated into a sparse linear programming problem. Then, a general framework based on the Bayesian model with independent Laplace prior was proposed to guarantee the sparseness and accuracy of identification results after analyzing influences of different prior distributions. At the same time, a three-stage hierarchical method was designed to resolve the puzzle that the Laplace distribution is not conjugated to the normal distribution. Last, the variational Bayesian was introduced to improve the efficiency of the network reconstruction task. The high accuracy and robust properties of the proposed method were verified by conducting both general synthetic network and real network identification tasks based on the evolutionary game dynamic. Compared with other five classical algorithms, the numerical experiments indicate that the proposed model can outperform these methods in both accuracy and robustness.

The prevalence and independent influencing factors of obesity and underweight in patients with schizophrenia: a multicentre cross-sectional study.

BACKGROUND: Few studies have investigated the weight of patients with schizophrenia in China. OBJECTIVE: The aim of this study was to analyse the prevalence, clinical characteristics and influencing factors of obesity and underweight in patients with chronic schizophrenia in China. METHODS: A total of 325 patients with schizophrenia and 172 sex- and age-matched healthy controls from the community were recruited. Socio-demographic data and laboratory measurements were collected for all subjects. Using the Positive and Negative Syndrome Scale (PANSS), we evaluated the psychiatric symptoms of patients with schizophrenia. According to the body mass index (BMI) criteria in China, BMI ≥ 28 kg/m2 indicates obesity, and BMI < 18.5 kg/m2 indicates underweight. RESULTS: Of the patients with schizophrenia, 16.3% were obese, and 6.8% were underweight; 11.0% of the healthy controls were obese, and 3.5% were underweight. There was no difference between the two groups in the prevalence of obesity and underweight. After controlling for relevant variables, the obesity rate remained non significant, but the underweight rate appeared to be different. The multinomial regression analysis revealed that among the patients with schizophrenia, female sex, triglyceride level and LDL level were independent risk factors for obesity and that HDL level was an independent protective factor against obesity. In contrast, male sex and HDL level were independent risk factors for underweight. CONCLUSION: We found that the patients with schizophrenia had an increased rate of underweight and some factors related to weight. LEVEL OF EVIDENCE: Level V, descriptive study.

Short-term effects of air pollution on cause-specific mental disorders in three subtropical Chinese cities.

BACKGROUND: The effects of ambient air pollution on specific mental disorders are rarely studied, and the reported results are inconsistent. OBJECTIVE: To assess the short-term effect of ambient air pollution on the morbidity of mental disorders in three subtropical Chinese cities. METHODS: Daily concentrations of air pollution were averaged from 19 fixed monitoring stations across each city, and data on patients were collected from three psychiatric specialty hospitals. A time-series study combined with a generalized additive Poisson model was conducted to investigate the association between air pollution and mental disorders. The exposure-response relationships were explored and stratified analyses by age and sex were conducted. RESULTS: A total of 1,133,220 outpatient visits were recorded in three subtropical cities (Huizhou, Shenzhen, and Zhaoqing). The number of daily outpatient visits for mental disorders increased with higher air pollutant (PM2.5, PM10, SO2 and NO2) concentrations, and the effect of NO2 appeared to be consistently significant across the three cities, with excess risk (ER) of 4.45% (95% CI: 2.90%, 6.04%) in Huizhou, 7.94% (95% CI: 6.28%, 9.62%) in Shenzhen, and 2.19% (95% CI: 0.51%, 3.89%) in Zhaoqing, respectively, at lag03. We also observed significant effect of PM2.5 at lag0 (ER = 1.20%, 95% CI: 0.28%, 2.13%), PM10 at lag0 (ER = 0.99%, 95% CI: 0.36%, 1.62%), and SO2 at lag0 (ER = 10.74%, 95% CI: 3.20%, 18.84%) in Shenzhen. For specific mental disorders, significant associations were found in all the air pollutants except between SO2 and affective disorder and between PM2.5 and schizophrenia. In addition, we found that air pollution exhibited stronger effects for males and adults (≥18 years). CONCLUSION: Acute exposure to air pollution, especially NO2, might be an important trigger of mental disorders.

Carbon and oxygen isotopic analyses of calcite in calcite-dolomite mixtures: Optimization of selective acid extraction.

RATIONALE: Carbon and oxygen isotopic compositions of individual minerals in calcite-dolomite mixtures could be measured using a selective acid extraction technique based on the different reaction rates of calcite and dolomite with phosphoric acid. However, poor accuracies of calcite are usually obtained when mixtures of low calcite content are analyzed, which may result in incorrect conclusions. To overcome this shortcoming, improvements are needed in accuracy and precision when using the technique with mixtures of low calcite content. METHODS: Calcite and dolomite were analyzed under different reaction conditions using a GV Isoprime™ II isotope ratio mass spectrometer equipped with a dual inlet (DI) and an automatic sampler, to study the relationships between isotopic compositions and CO2 yield, and reaction time and temperature. Artificial mixtures with varied calcite:dolomite ratios were prepared, and their calcite isotopic compositions were determined with the aim of optimizing analytical conditions. RESULTS: The CO2 yields and isotopic compositions of calcite and dolomite were obtained on-line under different experimental conditions using DI-isotope ratio mass spectrometry (DI-IRMS). Based on the analysis results of artificial mixtures, the reaction conditions for analyzing isotopic composition of calcite in mixtures were optimized as follows. Samples should have grain sizes of 75-80 µm; and samples with calcite contents of >50%, 30-50% and < 30% should be reacted with acid for 10-15 min at 50°C, 6 min at 50°C and 45 min at 25°C, respectively. CONCLUSIONS: Optimized analytical conditions are suggested for the determination of the isotopic compositions of calcite in mixtures with improved accuracy by an automatic selective acid extraction technique.

Study protocol for a parallel-group, double-blinded, randomized, controlled, noninferiority trial: the effect and safety of hybrid electroconvulsive therapy (Hybrid-ECT) compared with routine electroconvulsive therapy in patients with depression.

BACKGROUND: Electroconvulsive therapy (ECT) is the most rapid and effective treatment for patients with depression, ECT can achieve remarkable antidepressant effects in the initial 3-4 sessions, but significant side effects limit its use. However, recent low-charge electrotherapy (LCE) studies have demonstrated antidepressant or antipsychotic effects with significantly fewer side effects. The aim of this study is to propose a novel two-step charge set strategy for ECT treatment, referred to as Hybrid-ECT, to decrease side effects by using a low charge while preserving treatment efficacy. METHODS/DESIGN: A randomized, double-blinded, standard-controlled, parallel-group design will be carried out. We plan to enroll 112 inpatients diagnosed with depression (unipolar or bipolar) and randomly assign them to conventional ECT (control group) or to Hybrid-ECT (treatment group, 3 ECT sessions followed by LCE sessions (approximately 2.8 joules per session)). We will evaluate participants across a wide variety of domains including clinical symptoms, cognitive, psychological and functional metrics. We will also perform magnetic resonance imaging (MRI) and event-related potential (ERPs) assessments during treatment to explore brain function differences between ECT and LCE. DISCUSSION: This research proposes a simple but completely novel ECT strategy that aims to rapidly relieve depressive symptoms and minimize side effects. The mechanism of ECT and LCE will be further discussed. TRIAL REGISTRATION: Chinese Clinical Trial Registry, Number: ChiCTR1900022905 (Registration date: April 30, 2019).

Robust network structure reconstruction based on Bayesian compressive sensing.

Complex network has proven to be a general model to characterize interactions of practical complex systems. Recently, reconstructing the structure of complex networks with limited and noisy data attracts much research attention and has gradually become a hotspot. However, the collected data are often contaminated by unknown outliers inevitably, which seriously affects the accuracy of network reconstruction. Unfortunately, the existence of outliers is hard to be identified and always ignored in the network structure reconstruction task. To address this issue, here we propose a novel method which involves the outliers from the Bayesian perspective. The accuracy and the robustness of the proposed method have been verified in network reconstruction with payoff data contaminated with some outliers on both artificial networks and empirical networks. Extensive simulation results demonstrate the superiority of the proposed method. Thus, it can be concluded that since the proposed method can identify and get rid of outliers in observation data, it is conducive to improve the performance of network reconstruction.

Occurrence and regression of BK polyomavirus associated carcinoma: a clinical and next-generation sequencing study.

Low-level BK polyomavirus (BKPyV) shedding is seen in at least 10% of seropositive immunocompetent adults. Moreover, BKPyV infection is highly prevalent amongst immunocompromised populations, yet little is known on its relationship with malignancy. We studied a female patient with BKPyV-associated and donor-derived de novo high-grade sarcomatoid urothelial carcinoma developed 8 years after kidney transplantation from a male donor. Through whole-genome sequencing, we discovered integration of genotype IV BKPyV genome into the non-coding RNA (ncRNA) intronic region of human chromosome 18. The two breakpoints in the virus genome were located at the non-coding control region (NCCR) and large T antigen (TAg) coding region, respectively. Nevertheless, the TAg was overexpressed. We, therefore, inferred that the BKPyV was clonally integrated into the human genome in the form of concatemers, facilitating the expression of the TAg. The patient presented with multiorgan metastases, which were reduced in size and number throughout the body after removal of the graft and cessation of immunosuppressants. The few remaining lesions located in the liver were identified, through biopsy to be necrotic tumor tissue with TAg detected; additionally, genomic sequencing of the liver mass found Y chromosome. In conclusion, we propose that integration of the BKPyV genome is closely related to oncogenesis in this patient; while oncogenesis occurred when host immunity was impaired, recovery of the patient's native immunity effectively curbed viral replication and eliminated the metastatic lesions.

Peptides derived from the integrin ß cytoplasmic tails inhibit angiogenesis.

BACKGROUND: Integrins are essential regulators of angiogenesis. However, the antiangiogenic potential of peptides derived from the integrin cytoplasmic tails (CT) remains mostly undetermined. METHODS: Here we designed a panel of membrane-penetrating peptides (termed as mßCTPs), each comprising a C-terminal NxxY motif from one of the conserved integrin ß CTs, and evaluated their antiangiogenic ability using both in vitro and in vivo approaches. RESULTS: We found that mß3CTP, mß5CTP and mß6CTP, derived respectively from the integrin ß3, ß5 and ß6 CTs, but not others, exhibit antiangiogenic ability. Interestingly, we observed that the integrin ß3, ß5 and ß6 CTs but not others are able to interact with ß3-endonexin. In addition, the antiangiogenic core in mß3CTP is identical to a previously identified ß3-endonexin binding region in the integrin ß3 CT, indicating that the antiangiogenic mßCTPs may function via their binding to ß3-endonexin. Consistently, knockdown of endogenous ß3-endonexin in HUVECs significantly suppresses tube formation, suggesting that ß3-endonexin is proangiogenic. However, neither treatment with the antiangiogenic mßCTPs nor knockdown of endogenous ß3-endonexin affects integrin-mediated HUVEC adhesion and migration, indicating that their antiangiogenic effect may not rely on directly regulating integrin activity. Importantly, both treatment with the antiangiogenic mßCTPs and knockdown of endogenous ß3-endonexin in HUVECs inhibit VEGF expression and cell proliferation, thereby providing mechanistic explanations for the functional consequences. CONCLUSION: Our results suggest that the antiangiogenic mßCTPs can interact with ß3-endonexin in vascular endothelial cells and suppress its function in regulating VEGF expression and cell proliferation, thus disclosing a unique pathway that may be useful for developing novel antiangiogenic strategies.

Genetic diagnosis of polycystic kidney disease, Alport syndrome, and thalassemia minor in a large Chinese family.

Polycystic kidney disease (PKD) and Alport syndrome (AS) are serious inherited disorders associated with renal disease, and thalassemia is a hereditary blood disease with a high prevalence in south China. Here, we report an exceptional PKD coincidence of thalassemia minor and AS (diagnosed genetically) in a large Chinese family. Whole genome next-generation sequencing (NGS) was performed on the proband, and all family members underwent clinical evaluation. Sanger sequencing was used to validate the mutations distinguished by NGS. The pathogenic potential of the variants were evaluated by Polymorphism Phenotyping v2 (PolyPhen-2), Sorting Intolerant From Tolerant (SIFT) algorithm, and MutationTaster. Immunohistochemical, Western blot, immunofluorescent, and TdT-mediated dUTP nick-end labeling (TUNEL) analyses were performed to investigate polycystin 1 (PC1) expression, and cell proliferation and apoptosis in kidney tissues from the proband and normal control. A novel frameshift polycystic kidney disease 1 (PKD1) mutation (c.3903delC, p.A1302Pfs) was identified to be responsible for renal disease in this family. PC1 expression, and cell proliferation and apoptosis were significantly increased in the kidney tissues of the proband. Moreover, a deletion of approximately 19.3 kb of DNA with α-globin genes (_ _SEA) was associated with thalassemia minor in the family. In addition, a collagen type IV α 5 chain (COL4A5) variant (c.2858G>T, rs78972735), annotated as a pathogenic mutation in dbSNP and human gene mutation database (HGMD), was found in four family members with no clinical traits of AS. A novel pathogenic PKD1 mutation (c.3903delC) and (_ _SEA) thalassemia deletion were found to be responsible for the clinical symptoms in this family. The reported pathogenic COL4a5 variant (c.2858G>T, rs78972735) was not pathogenic alone.

Exploring the thermostable properties of halohydrin dehalogenase from Agrobacterium radiobacter AD1 by a combinatorial directed evolution strategy.

As a crucial factor for biocatalysts, protein thermostability often arises from a combination of factors that are often difficult to rationalize. In this work, the thermostable nature of halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) was systematically explored using a combinatorial directed evolution approach. For this, a mutagenesis library of HheC mutants was first constructed using error-prone PCR with low mutagenesis frequency. After screening approximately 2000 colonies, six mutants with eight mutation sites were obtained. Those mutation sites were subsequently combined by adopting several rounds of iterative saturation mutagenesis (ISM) approach. After four rounds of saturation mutagenesis, one best mutant ISM-4 with a 3400-fold improvement in half-life (t 1/2) inactivation at 65 °C, 18 °C increase in apparent T m value, and 20 °C increase in optimum temperature was obtained, compared to wild-type HheC. To the best of our knowledge, the mutant represents the most thermostable HheC variant reported up to now. Moreover, the mutant was as active as wild-type enzyme for the substrate 1,3-dichloro-2-propanol, and they remained most enantioselectivity of wild-type enzyme in the kinetic resolution of rac-2-chloro-1-phenolethanol, exhibiting a great potential for industrial applications. Our structural investigation highlights that surface loop regions are hot spots for modulating the thermostability of HheC, the residues located at these regions contribute to the thermostability of HheC in a cooperative way, and protein rigidity and oligomeric interface connections contribute to the thermostability of HheC. All of these essential factors could be used for further design of an even more thermostable HheC, which, in turn, could greatly facilitate the application of the enzyme as a biocatalyst.

[Characteristics of urinary tract infection in kidney transplant recipients and non-recipient patients].

OBJECTIVE: To compare the characteristics of urinary tract infection (UTI) between kidney transplant recipients and non-recipient patients. METHODS: Forty-nine kidney transplant recipients with UTI (69 episodes) and 401 non-recipient patients with UTI (443 episodes) admitted in Nanfang Hospital from January 2003 to August 2014 were enrolled in the study. The characteristics of UTI were compared between two groups. RESULTS: In both groups of UTI, female patients comprised a greater proportion (63.3% and 58.6%) and Escherichia coli was the most common pathogen isolated (37.7% and 34.1%). However, the infection rate of Klebsiella pneumonia in recipients was higher than that in non-recipients (11.6% vs 3.2%, P= 0.001), while the infection rate of Candida albicans was lower (1.5% vs 11.3%, P=0.008) than that in non-recipients. Recipients were likely to develop antibiotic resistance and with a higher recurrence rate than non-recipient patients (38.8% vs 16.7%, P<0.001). Compared to non-recipient UTI patients, the symptoms of urinary irritation in recipient UTI patients were more common. There was higher percentage of neutrophil granulocyte (72.65% ± 1.90% vs 68.59% ± 0.73%, P=0.048), lower proportion of lymphocytes (17.73% ± 1.27% vs 21.28% ± 0.61%, P=0.037), and less platelets [(187.64 ± 10.84) × 10(9)/L vs (240.76 ± 5.26) × 10(9)/L, P<0.01] in recipients than in non-recipient UTI patients. CONCLUSION: These results indicate that the characteristics of UTI in kidney transplantation recipients and non-recipients patients are different.