Inhibition of IGF2BP1 attenuates the progression of endometriosis through PTBP1.

INTRODUCTION: Endometriosis (EMs), manifested by pain and infertility, is a chronic inflammatory disease. The precise pathophysiology of this disease remains uncertain. Insulin-like growth factor-2 mRNA-binding protein 1 (IGF2BP1) and polypyrimidine tract-binding protein 1 (PTBP1) have both been found to regulate proliferation, apoptosis, and invasion. This study aimed to investigate the effects of IGF2BP1/PTBP1 in treating EMs. MATERIALS AND METHODS: qRT-PCR and western blotting were employed to quantify IGF2BP1 and PTBP1 expression in six patients with EMs (mean age 33.83 years). The correlation analysis, STRING database prediction, and RNA immunoprecipitation were utilized to identify the relationship between IGF2BP1 and PTBP1. Ectopic endometrial volume, weight, HE staining, and IGF2BP1 silencing were utilized to estimate the effects of IGF2BP1 in EMs model rats. qRT-PCR, CCK-8, 5-ethynyl-2'-deoxyuridine (EDU) labeling, Transwell assay, and flow cytometry were utilized to assess the effects of IGF2BP1/PTBP1 on the proliferation, migration, invasion, and apoptosis of ectopic endometrial stromal cells (eESCs). Furthermore, western blotting was employed to evaluate expressions of PCNA, VEGF, and E-cadherin in EMs rats and eESCs. RESULTS: The mRNA and protein levels of IGF2BP1 and PTBP1 in the ectopic and eutopic endometrium of EMs patients were significantly increased. RNA immunoprecipitation revealed a close interaction of IGF2BP1 with PTBP1. Additionally, the endometrial volume, weight, and histopathologic scores in rats were significantly reduced after IGF2BP1 silencing. IGF2BP1 silencing also decreased the expression of PCNA and VEGF, and increased E-cadherin expression in endometrial tissues of EMs rats. Moreover, IGF2BP1 silencing inhibited proliferation, migration, and invasion and promoted apoptosis through PTBP1 in eESCs. CONCLUSIONS: IGF2BP1 exhibits potential beneficial properties in the management of EMs by interacting with PTBP1, thereby highlighting IGF2BP1 as a promising therapeutic target for EMs.

Preservation of Simmental bull sperm at 0°C in Tris dilution: effect of dilution ratio and long-distance transport.

OBJECTIVE: This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0°C, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. METHODS: In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0°C for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. RESULTS: The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0°C gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0°C for 1 to 6 days. There was no significant difference (p>0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p>0.05). CONCLUSION: This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0°C conditions. Additionally, it is feasible to distribute semen regionally.

Effect of comprehensive nursing methods in postoperative ICU of children with CHD.

To explore effect of comprehensive nursing in postoperative ICU of children with CHD. The subjects were 50 cases of children with CHD treated in our hospital: 25 cases in the control group: routine nursing, and 25 cases in the observation group: comprehensive nursing intervention. The effective rate of 92.00% in the observation group was significantly higher. The serum-free calcium value (1.07 ± 0.11) mmol/L of the observation group on the first day after surgery was significantly lower, and the observation group's creatine phosphate, the daily average dosage of creatine phosphate per unit body weight was significantly higher. 96.00% of patients in the observation group were significantly higher in nursing satisfaction. The complication rate of 8.00% in observation group was significantly lower. In order to successfully complete the operation schedule and improve the postoperative recovery effect of children, high requirements are placed on nursing staff. The comprehensive nursing method used in the postoperative ICU of children with CHD can reduce the incidence of postoperative complications and improve nursing satisfaction.

Macrophage KLF15 prevents foam cell formation and atherosclerosis via transcriptional suppression of OLR-1.

BACKGROUND: Macrophage-derived foam cells are a hallmark of atherosclerosis. Scavenger receptors, including lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (OLR-1), are the principal receptors responsible for the uptake and modification of LDL, facilitating macrophage lipid load and the uptake of oxidized LDL by arterial wall cells. Krüppel-like factor 15 (KLF15) is a transcription factor that regulates the expression of genes by binding to the promoter during transcription. Therefore, this study aimed to investigate the precise role of macrophage KLF15 in atherogenesis. METHODS: We used two murine models of atherosclerosis: mice injected with an adeno-associated virus (AAV) encoding the Asp374-to-Tyr mutant version of human PCSK9, followed by 12 weeks on a high-fat diet (HFD), and ApoE-/-- mice on a HFD. We subsequently injected mice with AAV-KLF15 and AAV-LacZ to assess the role of KLF15 in the development of atherosclerosis in vivo. Oil Red O, H&E, and Masson's trichome staining were used to evaluate atherosclerotic lesions. Western blots and RT-qPCR were used to assess protein and mRNA levels, respectively. RESULTS: We determined that KLF15 expression was downregulated during atherosclerosis formation, and KLF15 overexpression prevented atherosclerosis progression. KLF15 expression levels did not affect body weight or serum lipid levels in mice. However, KLF15 overexpression in macrophages prevented foam cell formation by reducing OLR-1-meditated lipid uptake. KLF15 directly targeted and transcriptionally downregulated OLR-1 levels. Restoration of OLR-1 reversed the beneficial effects of KLF15 in atherosclerosis. CONCLUSION: Macrophage KLF15 transcriptionally downregulated OLR-1 expression to reduce lipid uptake, thereby preventing foam cell formation and atherosclerosis. Thus, our results suggest that KLF15 is a potential therapeutic target for atherosclerosis.

Identification of GINS1 as a potential prognostic biomarker for sarcoma using bioinformatic analysis.

Background: The GINS complex is related to cancer development, invasion, and poor prognosis in multiple tumors. In the study, we attempted to investigate the prognostic value of GINS1 in sarcoma patients. Methods: We analyzed GINS1 expression using Tumor IMmune Estimation Resource (TIMER) 2.0, Gene Expression Omnibus (GEO; GSE21122, GSE39262, and GSE21050), and The Cancer Genome Atlas (TCGA) databases. The prognostic value of GINS1 was explored using the survival and survminer packages of R. Genetic alteration analysis was investigated using cBioPortal. The Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) R script was used for the immunocyte infiltration analysis. MicroRNAs (miRNAs) targeting GINS1 were predicted using GEO (GSE69470) and MicroRNA Target Prediction Database (miRDB). Results: We found that GINS1 was overexpressed in sarcoma, especially in metastatic samples, and was associated with a worse prognosis. High GINS1 expression was a poor prognostic indicator for sarcoma patients. Moreover, GINS1 alteration was associated with worse survival of sarcoma patients. Immune infiltration analysis indicated that GINS1 expression was correlated with the infiltration of M0 and M2 macrophages in sarcoma. Finally, the miRNA hsa-miR-376a-3p was identified to potentially regulate GINS1 in sarcoma. Conclusions: These results indicate that GINS1 may be a promising prognostic biomarker and therapeutic target for sarcoma.

Mechanism of Procyanidin B2 in the Treatment of Chronic Myeloid Leukemia Based on Integrating Network Pharmacology and Molecular Docking.

OBJECTIVE: To study the pharmacological mechanism of procyanidin B2 (PCB2) on chronic myeloid leukemia (CML) by integrating network pharmacological methods systematically. METHODS: Firstly, the potential target genes of PCB2 were predicted by the pharmacological database and analysis platform (TCMSP and Pharmmapper). Meanwhile, the relevant target genes of CML were collected from GeneCards and DisGene. Pooled data were collected to screen for common target genes. Furthermore, the above intersection genes were imported into the String website to construct a protein-protein interaction (PPI) network, and the Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were further analyzed. Besides, molecular docking was performed to verify the possible binding conformation between PCB2 and candidate targets. Finally, MTT and RT-PCR experiments of K562 cells were performed to verify the above results of network pharmacology. RESULTS: A total of 229 PCB2 target genes were retrieved, among which 186 target genes had interaction with CML. The pharmacological effects of PCB2 on CML were related to some important oncogenes and signaling pathways. The top ten core targets predicted by Network Analysis were as follows: AKT1, EGFR, ESR1, CASP3, SRC, VEGFA, HIF1A, ERBB2, MTOR, and IGF1. Molecular docking studies confirmed that hydrogen bonding was the main interaction force of PCB2 binding targets. According to the molecular docking score, the following three target proteins were most likely to bind to PCB2: VEGFA (-5.5 kcal/mol), SRC (-5.1 kcal/mol), and EGFR (-4.6 kcal/mol). After treatment of PCB2 for 24h, mRNA expression levels of VEGFA and HIF1A decreased significantly in K562 cells. CONCLUSION: Through integrating network pharmacology combined with molecular docking, the study revealed the potential mechanism of PCB2 anti-chronic myeloid leukemia.

A Sucrose Fatty Acid Ester-mixed Emulsifier as Stabilizer of Emulsion Systems.

In the present study, the mesomorphic phase and internal structure of sucrose fatty acid ester (S1670) with cetyl alcohol mixed in different proportions was studied and assessed, within common emulsion system of cosmetics and pharmaceuticals. To characterize the system's colloidal structure the following physicochemical techniques were used: polarization microscopy, small- and wide-angle X-ray scattering (SAXS and WAXS), differential scanning calorimetry (DSC), continual and oscillatory rheology and cryogenic scanning electron microscopy (cryo-SEM). Ultimately, it was well established that there was a predominant and stable α-crystalline gel structure, formed with sucrose fatty acid ester (S1670) and cetyl alcohol, in the pseudo quarter-phase system (including squalane and aqua). In addition, the optimum ratio for α-crystalline gel formation was found to be around 61.50:38.50 (S1670 to Cetyl alcohol). Moreover, as it deviates from the critical point within a controllable range, although identical α-gel (α-form hydrated crystal) continues to exist, yet the strength and proportion in whole sample decrease with the expansion of deviation. The decent stability (which was derived from the dismatch between the lipophilic chains and the intensive hydration of -OH from S1670) and the agreeable rheological properties of the above α-gel suggest wide application in cosmetics and pharmacy in the future.

The coordinated impacts of agricultural insurance and digital financial inclusion on agricultural output: Evidence from China.

The coordinated development of agricultural insurance and digital financial inclusion is crucial to agricultural production, previous research on the subject is unclear. Based on the data of some provinces in China from 2011 to 2019, the entropy method is used in this paper to construct indexes measuring the development of agricultural insurance and digital financial inclusion. Their coupling coordination index is calculated, and the basic characteristics of the coupling coordination degree are analyzed. The influence of the coupling coordination degree of agricultural insurance and digital financial inclusion on agricultural output is empirically analyzed by constructing a regression model. The results show that the coupling coordination degree of agricultural insurance and digital financial inclusion significantly improves farmers' agricultural output, and the promotion effect is more prominent in eastern China and mountainous areas. And the threshold effect analysis show that there is a nonlinear relationship between the coupling coordination degree of agricultural insurance and digital financial inclusion on agricultural output. The conclusion of this paper provides a theoretical basis and empirical evidence for the coordinated development of rural financial system and agricultural construction.

Sources and historical records of metal(loid)s in river sediments from an agricultural tributary of Tuojiang river: Evidence from geochemical analyses and Pb isotope.

Clarifying the sources and historical variation of metal(loid)s in agricultural river sediments is vital for watershed contamination control and environmental improvements. In this study, a systematical geochemical investigation of Pb isotopic characteristics and the spatial-temporal distribution of metal(loid)s abundances was conducted to delineate the origins of metal(loid)s (Cd, Zn, Cu, Pb, Cr, and As) in sediments from an agricultural river, Sichuan Province, Southwest China. The results showed significant enrichment of Cd and Zn in the whole watershed, with substantial anthropogenic contributions of 86.1% and 63.1% for the surface sediments, and 79.1% and 67.9% for the core sediments, respectively. As was mainly derived from natural sources. Cu, Cr, and Pb were originated from the mixing sources of natural and anthropogenic processes. The anthropogenic origin of Cd, Zn, and Cu in the watershed was closely correlated with agricultural activities. The profile of EF-Cd and EF-Zn displayed an increasing trend from the 1960s-1990s, and then kept a high value, which was consistent with the development of national agricultural activities. Pb isotopic signatures suggested multiple sources of the anthropogenic Pb contamination, including industrial/sewage discharge, coal combustion, and vehicle exhaust. The average anthropogenic 206Pb/207Pb ratio (1.1585) approximated that of local aerosols (1.1660), suggesting aerosol deposition was a crucial pathway of anthropogenic Pb input to sediment. Furthermore, the anthropogenic Pb percentages (mean of 52.3 ± 10.3%) from the EF approach were in line with that from the Pb isotopic method (mean of 45.5 ± 13.3%) for sediments under intense anthropogenic impacts.

Effects of Acupuncture on Cancer Pain in Animal Intervention Studies: A Systematic Review and Quality Assessment.

BACKGROUND: Preclinical studies often provide the evidence base for clinical studies. However, the design and reporting of preclinical trial results are inadequate, resulting in poor reproducibility and clinical translatability. We aimed to systematically evaluate the methodology and reporting quality of animal studies of acupuncture for cancer pain. METHODS: About 7 databases were searched for animal research articles on acupuncture for cancer pain from the beginning of the database to January 31, 2022. ARRIVE guidelines, STRICTA, and SYRCLE risk of bias tools were used to assess the reporting quality and risk of bias of the selected studies. RESULTS: A total of 18 studies were evaluated. Of the 22 items on the SYRCLE tool, only 6 items had a positive reporting rate of more than 50%. Of the 39 items in the ARRIVE guidelines, 14 were rated excellent, and the least frequently reported checklist items were 7. Out of the 17 STRICTA checklist items analyzed, 10 were considered appropriately reported in more than 80% of the studies, while 4 were correctly reported in less than 20%. CONCLUSIONS: Some crucial points in the design, implementation, and reporting of the experiments included in the study were not well developed, which could significantly affect the clarity, reproducibility, and translatability of the experiments. There is a need to fully implement scientific tool guidelines for future experimental studies in order to improve the quality of preclinical studies and facilitate effective translation of their results to the clinic.

Observation of Anesthetic Effect of Dexmedetomidine Combined With Intraspinal Anesthesia in Hip Arthroplasty and its Effect on Postoperative Delirium and Stress Response.

Objective: To observe the anesthetic effect of dexmedetomidine combined with spinal anesthesia in hip arthroplasty, and to analyze the effects of dexmedetomidine on postoperative stress response, incidence of delirium, immune function and inflammatory indicators. Methods: A total of 42 patients who underwent hip replacement in our hospital from March 2020 to June 2021 were selected as the research subjects and randomly divided into the control group and the observation group, 21 cases in each group. The control group was given intraspinal anesthesia, and the observation group was given dexmedetomidine on this basis. The onset time and maintenance time of sensory and motor nerve block were recorded. Stress response indexes [cortisol (Cor), blood glucose (Glu), adrenaline (E), noadrenaline (NE)], T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+), inflammatory indexes [tumor necrosis factor -α (TNF-α) and interleukin-6 (IL-6)] were detected before and after operation, and the incidence of postoperative delirium in both groups was recorded. Results: The onset time of sensory nerve block and motor block in the observation group were lower than those in the control group, and the retention time of sensory nerve block and motor nerve block were higher than those in the control group (P < 0.05). After surgery, the levels of Cor, Glu, E and NE in the observation group were lower than those in the control group (P < 0.05). After surgery, the incidence of postoperative delirium in the observation group (4.79%) was lower than that in the control group (28.57%) (P < 0.05). After surgery, the levels of CD3+, CD4+, CD8+, and CD4+/CD8+ in the observation group were higher than those in the control group (P < 0.05). After surgery, the levels of TNF-α and IL-6 in the observation group were lower than those in the control group (P < 0.05). Conclusion: The combined use of dexmedetomidine and intraspinal anesthesia has good anesthesia effect in hip joint replacement, which can greatly reduce the stress response of patients, reduce the incidence of postoperative delirium, and effectively restore the immune function of patients, reduce the level of inflammatory response, and has high clinical application value.

Angiopoietin-like 3 (ANGPTL3) drives cell proliferation, migration and angiogenesis in cervical cancer via binding to integrin alpha v beta 3.

Angiopoietin-like 3 (ANGPTL3) has been uncovered to play an oncogenic role in several kinds of human malignancies. Nevertheless, whether ANGPTL3 functions in cervical cancer (CC) has not yet been reported. This paper is intended to explore the impact of ANGPTL3 on CC cells and elucidate the potential mechanism. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were performed to analyze the ANGPTL3 expression. Western blot was also performed to examine integrin αvß3 protein level. Cell proliferation was evaluated by MTT assay, EdU staining and Western blot analysis. In addition, the migratory and invasive abilities of cells were, respectively, estimated by wound healing and transwell assays. Tube formation assay was performed to determine endothelial cell angiogenesis. Levels of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 2 (VEGFR2) were measured by ELISA. As a result, ANGPTL3 expression was significantly higher in CC cells relative to that in normal cervical cells. Silencing of ANGPTL3 suppressed cell proliferation, migration and invasion. Besides, downregulation of ANGPTL3 inhibited human umbilical vein endothelial cell (HUVEC) angiogenesis and repressed protein level of integrin alpha v beta 3 (αvß3). Upregulation of αvß3 offsets the inhibitory effect of ANGPTL3 on proliferation, migration and invasion in CC cells. Upregulated expression of αvß3 promoted blood vessel formation and secretions of VEGF and VEGFR2. In conclusion, ANGPTL3 silencing may serve as a tumor suppressor in CC through integrin αvß3, which provides a potentially novel therapeutic target for patients with CC.

LncRNA SNHG15 Knockdown Protects Against OGD/R-Induced Neuron Injury by Downregulating TP53INP1 Expression via Binding to miR-455-3p.

Cerebral ischemia-reperfusion (I/R) injury is the common symptom of ischemic stroke, which poses a heavy burden to human health. Long non-coding RNA (lncRNA) is indicated to be a critical regulator in cerebral ischemia. This study aims to reveal the effects of lncRNA small nucleolar RNA host gene 15 (SNHG15) on oxygen-glucose deprivation and reoxygenation (OGD/R)-induced neuron injury and underlying mechanism. The expression levels of SNHG15, microRNA-455-3p (miR-455-3p) and tumour protein p53 inducible nuclear protein 1 (TP53INP1) mRNA were determined by quantitative real time polymerase chain reaction in P12 cells. The protein levels of TP53INP1, cleaved caspase-3, caspase-3, B-cell lymphoma-2 and BCL2-associated x protein (Bax) were detected by western blot in P12 cells. Cell viability and apoptosis were revealed by cell counting kit-8 assay and flow cytometry analysis, respectively, in P12 cells. Caspase-3 activity, the levels of tumor necrosis factor-α and interleukin-1ß and the production of reactive oxygen species (ROS) were severally determined by caspase-3 activity assay, Enzyme-linked immunosorbent assay and ROS detection assay in P12 cells. The binding relationship between miR-455-3p and SNHG15 or TP53INP1 was predicted by starbase online database, and identified by dual-luciferase reporter, RNA pull-down or RNA immunoprecipitation assay. SNHG15 expression and the mRNA and protein levels of TP53INP1 were dramatically upregulated, while miR-455-3p expression was apparently downregulated in OGD/R-induced PC12 cells. SNHG15 silencing hindered the effects of OGD/R treatment on cell viability, apoptosis, inflammation and oxidative in PC12 cells; however, these impacts were restored after miR-455-3p inhibitor transfection. Additionally, SNHG15 acted as a sponge of miR-455-3p and miR-455-3p bound to TP53INP1. SNHG15 contributed to OGD/R-induced neuron injury by regulating miR-455-3p/TP53INP1 axis, which provided a novel insight to study lncRNA-directed therapy in ischemia stoke.

Evaluation of seven rapid diagnostic tests for detection of hepatitis C virus antibodies in China.

Rapid diagnostic tests as an attractive alternative to enzyme immunoassay could identify hepatitis C virus (HCV) infected persons more expeditiously. The availability of high performing and quality-assured rapid diagnostic tests are essential to scale-up HCV screening. The study was undertaken to evaluate the performance of seven domestic HCV rapid diagnostic tests kits. The kits were evaluated by using HCV serum panels, including HCV basic panel, analytical specificity panel, mixed titre performance panel, characteristic panel, seroconversion panel, and genotype qualification panel. The results showed that clinical sensitivity, clinical specificity and analytical specificity of seven rapid diagnostic tests kits ranged from 94% (95% CI: 83.2-98.6) to 100% (95% CI: 91.5-100). Furthermore, specimens with HCV genotypes 1b, 2a, 3a, 4a, 5a, 6 could be detected by HCV rapid diagnostic tests kits, whereas specimens with genotypes 1a and 2b could not be detected. Additionally, most HCV rapid diagnostic tests kits had great performance in diagnosing different titres and/or different bands samples, but some low S/CO value specimens may not be fully detected by few rapid diagnostic test kits. In conclusion, seven HCV rapid diagnostic tests reagents presented high sensitivity, specificity, good anti-interference and detection ability of early infection, which could meet the requirements of clinical HCV antibody screening.

The glucagon-like peptide-1 (GLP-1) analog exenatide ameliorates intrauterine adhesions in mice.

OBJECTIVE: The purpose of the experiments in this study was to explore the effect of exenatide on intrauterine adhesions (IUAs) and to elucidate its mechanism to provide new ideas for the clinical treatment of IUAs. METHODS: In this study, an animal model of IUAs was established by double stimulation using mechanical curettage and inflammation. After modeling, the treatment group was injected subcutaneously with three doses of exenatide for two weeks. The model group was injected with sterile ultrapure water, and the sham operation group was treated the same as the normal group, except for the observation of abdominal wound changes. Two weeks later, all mice were sacrificed by cervical dysfunction. The obtained mouse uterine tissue was used for subsequent experimental detection, using HE and Masson staining for histomorphological and pathological analysis; qRT-PCR for the detection of TGF-ß1, α-SMA, and MMP-9 gene expression in uterine tissue; and western blotting analysis of TGF-ß1, α-SMA, and collagen 1 protein expression to verify whether exenatide has a therapeutic effect on IUAs in mice. RESULTS: In the high-dose exenatide treatment group, the endometrial glands significantly increased in size, and the deposition area of collagen fibers in the endometrial tissue was significantly reduced. We observed that the mRNA expression of TGF-ß1 and α-SMA in the endometrial tissue of IUAs mice in this group was significantly reduced, while the expression of MMP-9 was significantly increased. In addition, we found that the protein expression of TGF-ß1, α-SMA, and collagen 1 remarkably decreased after treatment with exenatide. CONCLUSION: Exenatide may reduce the deposition of collagen fibers in the uterus of IUAs mice and promote the proliferation of endometrial glands in mice.

Ablation of TMEM126B protects against oxygen-glucose deprivation/reoxygenation-induced injuries of PC12 cells via maintaining mitochondrial anti-apoptotic functions.

Ischemia reperfusion (I/R) injury is a key contributing factor to the pathogenic mechanism involved in cerebral infarction. Transmembrane protein 126b (TMEM126B), a mitochondrial complex I assembly factor, has been reported to have an intimate association with disease progression, but is little known in ischemia stroke. The present study was designed to explore the effects of TEME126B on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced neuronal PC12 cells. The mRNA level of TMEM126B was determined using qRT-PCR. The levels of ROS, MDA, and SOD, as well as inflammatory cytokines, were measured using corresponding commercial kits. Cell apoptosis rate was assayed by flow cytometry analysis, and the apoptosis-related proteins were measured using western blotting. ATP production measured by colorimetric reaction and mitochondrial membrane potential measured by JC-1 staining were conducted to determine mitochondrial dysfunction. The results showed that TMEM126B was upregulated upon I/R injury in vitro and in clinical, and was positively corrected with the degree of oxidative stress. TMEM126B knockdown significantly reduced oxidative stress and inflammation in OGD/R-induced PC12 cells. TMEM126B knockdown also attenuated cell apoptosis rate, accompanied with increased expressions of Bcl-2, XIAP and cleaved PARP-1, and decreased expressions of Bax, cleaved caspase 3 and cleaved caspase 9. Furthermore, TMEM126B knockdown exhibited cytoprotective roles through alleviating mitochondrial dysfunction, as assessed by ATP production and mitochondrial membrane potential. Collectively, this study indicates that TMEM126B knockdown protects against OGD/R-induced neuronal injuries through relieving oxidative stress, inflammation, apoptosis and mitochondria dysfunction, which provides a promising target for ischemic stroke treatment.

Shoutai pills improve the quality of oocytes exposed to the chemotherapeutic drug Hydroxyurea.

Hydroxyurea (HU), a DNA synthesis inhibitor, is one of the most common chemotherapeutic drugs that have been widely applied to treat a variety of cancers. HU treatment exhibits severe side effects including renal toxicity, skin toxicity and embryo-toxicity. However, the influence of HU on the female gamete development has not yet fully clarified. Here, we found that HU exposure induced the degeneration of activated follicles after primordial follicle stage, resulting in the depletion of the ovarian reserve. HU exposure also led to the oocyte meiotic maturation arrest via disrupting normal spindle assembly, chromosome alignment and kinetochore-microtubule attachment. Furthermore, exposure to HU impaired the dynamics of ovastacin and Juno, two critical fertilization regulators. Notably, we illustrated that Shoutai pills (STP), a traditional Chinese medicine drug that has been commonly used for the treatment of miscarriage in China, partially restored all of the defects of oocyte development resulting from HU exposure through inhibiting the occurrence of oxidative stress-induced apoptosis. Taken together, our data not only reveal the adverse impact of HU exposure on the female gamete development, but also provide an effective strategy to prevent it, potentially contributing to the improvement of the quality of oocytes from patients treated with HU.

An efficient method for simultaneously screening for HIV, syphilis, and HCV based on one dried blood spot sample.

OBJECTIVE: The aim of this study was to optimize the parameters of a dried blood spot (DBS)-based ELISA method to simultaneously screen for anti-HIV, anti-hepatitis C virus (HCV) and anti-treponema pallidum (TP) antibodies, and investigate the assay performance. METHODS: Experiments were performed to establish optimized parameters for a DBS-based ELISA method to simultaneously screen for anti-HIV, anti-HCV, and anti-TP antibodies. Then, 429 paired plasma and DBS samples were collected to evaluate the performance of the assay with optimized parameters. Plasma test results were used as the reference standard. RESULTS: The optimized assay conditions were: blood volume, 70-100 µL; DBS size, whole spot; eluent volume, 500 µL; eluent, PBS with 1‰ Tween20; elution time, 4 h; elution temperature, room temperature. The overall prevalence of HIV, HCV, and TP was 6.06% (95% confidence interval [CI]: 4.07-8.87%), 27.74% (95% CI: 23.60-32.28%), and 14.92% (95% CI: 11.75-18.73%). The clinical sensitivity of the assay for anti-HIV, anti-HCV, and anti-TP antibodies was 88.46%, 98.32%, and 92.19%, respectively. The assay was 100% specificity for each analyte. The assay positive predictive value for each analyte was 100%, and the negative predictive values were >98%. Of the 429 samples, 9 DBS results were different than the plasma results; in these samples the plasma signal-to-cutoff rations were low (range, 1.40-7.81). CONCLUSION: The DBS-based ELISA method demonstrated good performance for simultaneously screening for anti-HIV, anti-HCV, and anti-TP antibodies. DBS samples are a promising method to screen for HIV, HCV, and TP infections. IMPORTANCE: DBS samples are a promising alternative to plasma samples, with the advantages of good sample stability, smaller blood volume, simpler storage, and easier transport. DBS sample testing is used for the diagnosis of a wide variety of pathogens, including HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV). The performance of DBS samples for the serological diagnosis of HIV, HCV, and Treponema pallidum (TP) has been demonstrated to be good in many studies, and therefore DBS is regarded as an ideal choice to screen for HIV, HCV, and TP infections, especially in difficult-access or resource-limited settings. There is no generally accepted procedure for sample collection and processing of DBS samples and there are few studies on simultaneous detection of anti-HIV, anti-HCV and anti-TP using a single DBS.

Downregulation of transcription factor TCTP elevates microRNA-200a expression to restrain Myt1L expression, thereby improving neurobehavior and oxidative stress injury in cerebral palsy rats.

Transcription factors have already been proposed to work on some human diseases. Yet the role of translationally controlled tumor protein (TCTP) in cerebral palsy (CP) remains elusive. This study intends to examine the mechanism of TCTP on CP by regulating microRNA-200a (miR-200a).CP models of rats were established referring to the internationally recognized improved hypoxic ischemic encephalopathy modeling method. The neuroethology of rats, ultrastructure and pathological condition in brain tissues of rats were observed through several assays. The expression of TCTP, miR-200a, myelin transcription factor 1-like (Myt1L), tyrosine hydroxylase (TH) and inducible nitric oxide synthase (iNOS) along with apoptosis in brain tissues of rats was detected. The levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in brain tissues of rats were determined. The binding site between miR-200a and Myt1L was analyzed.TCTP and Myt1L were overexpressed and miR-200a was under-expressed in CP rats. Elevated miR-200a ameliorated neurobehavior of CP rats and pathological injury in brain tissues. Elevated miR-200a up-regulated TH, GSH, GSH-Px, and SOD levels, down-regulated iNOS, ROS, MDA, TNF-α, and IL-6 levels, and attenuated neuronal apoptosis in brain tissues of CP rats. Myt1L was a target gene of miR-200a.Altogether, our study suggested that diminution of transcription factor TCTP up-regulates miR-200a to limit Myt1L expression, thereby improving neurobehavior and oxidative stress injury in CP rats.

HIV DNA Measurement and Improved Detection of HIV Infection Among Men Who Have Sex with Men: A Strategic Implication.

Rapid test (RT) is the principal screening method in the HIV control practice. However, this method may lead to inaccurate detection, primarily due to the more than 4 weeks of window phase. In the present study, we performed a HIV DNA screening method to show its application prospects in men who have sex with men (MSM). From July 2017 to April 2018, we recruited 1,301 MSM from Beijing who were not previously diagnosed as HIV positive. Both HIV DNA detection and RT were performed. In total, 141 and 135 HIV-positive results were detected by DNA detection and RT, respectively. By repetitive and confirmative tests (Western blot), we verified that DNA detection detected 10 more true positives than RT and 4 false positives were corrected from RT. This represents 14 inaccurate RT results that were corrected by DNA measurement. Therefore, DNA measurement should be fully considered as a screening method in the detection of HIV among MSM in the future.