BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

Objective To investigate whether miR-15b-5p can alleviate airway inflammation in asthma by negatively regulating ubiquitin specific peptidase 9X (USP9X) to down-regulate the expression of phosphatidylinositol 4, 5-diphosphate 3-kinase catalytic subunit α/AKT serine/threonine kinase 1 (PIK3CA/AKT1) pathway. Methods USP9X was predicted to be a direct target of miR-15b-5p by using an online database (miRWalk), and the luciferase reporter gene assay was performed to verify it. Co-immunoprecipitation (CO-IP) was used to verify the direct binding between USP9X and PIK3CA and the role of USP9X and its small molecule inhibitor WP1130 in the deubiquitination of PIK3CA. C57 mice were randomly divided into Control group, OVA group, OVA combined with NC group and miR-15b-5p agomir group, with 10 mice in each group. BEAS-2B cells were induced with interleukin 13 (IL-13) and treated with miR-15b-5p mimic. HE, Masson, PAS, immunohistochemistry, immunofluorescence staining, flow cytometry, Western blot and quantitative real-time PCR(qRT-PCR) were performed. Results It was found that the administration of miR-15b-5p agomir and mimic could reduce peribronchial inflammatory cells and improve airway inflammation, and miR-15b-5p could target negative regulation of USP9X. USP9X could directly bind to PIK3CA and regulate PIK3CA level in a proteasome-dependent manner, and USP9X could deubiquitinate K29-linked PIK3CA protein. Down-regulation of USP9X could increase PIK3CA ubiquitination level. WP1130, a small molecule inhibitor of USP9X, has the same effect as knockdown of USP9X, both of which could increase the ubiquitination level of PIK3CA and reduce the protein level of PIK3CA. Conclusion The miR-15b-5p/USP9X/PIK3CA/AKT1 signaling pathway may provide potential therapeutic targets for asthma.
Animals ; MicroRNAs/metabolism* ; Asthma/pathology* ; Class I Phosphatidylinositol 3-Kinases/genetics* ; Ubiquitin Thiolesterase/metabolism* ; Proto-Oncogene Proteins c-akt/genetics* ; Mice ; Signal Transduction ; Mice, Inbred C57BL ; Humans ; Inflammation/genetics* ; Cell Line ; Female ; Male

BACKGROUND: The sirtuin family is well recognized for its crucial involvement in various cellular processes. Nevertheless, studies on its role in the human endometrium are limited. This study aimed to explore the expression and localization of the sirtuin family in the human endometrium, focusing on sirtuin 3 (SIRT3) and its potential role in the oxidative imbalance of the endometrium in polycystic ovary syndrome (PCOS). METHODS: Endometrial specimens were collected from both patients with PCOS and controls undergoing hysteroscopy at the Center for Reproductive Medicine, Peking University Third Hospital, from July to August 2015 and used for cell culture. The protective effects of SIRT3 were investigated, and the mechanism of SIRT3 in improving endometrial receptivity of patients with PCOS was determined using various techniques, including cellular bioenergetic analysis, small interfering ribonucleic acid (siRNA) silencing, real-time quantitative polymerase chain reaction, Western blot, immunofluorescence, immunohistochemistry, and flow cytometry analysis. RESULTS: The sirtuin family was widely expressed in the human endometrium, with SIRT3 showing a significant increase in expression in patients with PCOS compared with controls ( P <0.05), as confirmed by protein and gene assays. Concurrently, endometrial antioxidant levels were elevated, while mitochondrial respiratory capacity was reduced, in patients with PCOS ( P <0.05). An endometrial oxidative stress (OS) model revealed that the downregulation of SIRT3 impaired the growth and proliferation status of endometrial cells and reduced their receptivity to day 4 mouse embryos. The results suggested that SIRT3 might be crucial in maintaining normal cellular state by regulating antioxidants, cell proliferation, and apoptosis, thereby contributing to enhanced endometrial receptivity. CONCLUSIONS Our findings proposed a significant role of SIRT3 in improving endometrial receptivity in patients with PCOS by alleviating OS and regulating the balance between cell proliferation and apoptosis. Therefore, SIRT3 could be a promising target for predicting and improving endometrial receptivity in this patient population.
Humans ; Female ; Polycystic Ovary Syndrome/metabolism* ; Endometrium/metabolism* ; Sirtuin 3/genetics* ; Oxidative Stress/genetics* ; Adult ; Animals ; Mice ; Apoptosis/physiology* ; Immunohistochemistry ; Cell Proliferation/physiology*

Objective:To analyze the genetic evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) of influenza B virus in Tongling during 2019-2022 surveillance years.Methods:Twenty-two strains of Victoria influenza B virus isolated from our laboratory during 2019-2022 were selected for whole genome sequencing. The sequence comparison and phylogenetic analysis were conducted by using bioinformatic analysis software.Results:During 2019-2022, seasonal influenza in Tongling City was predominantly caused by influenza B Victoria lineage viruses, which fell within the V1A.3 branch. Among these, 14 strains isolated in the 2021-2022 season were further classified into the V1A.3a.2 sub-branch. Compared with vaccine strains, multiple amino acid mutation sites were detected in both HA and NA proteins of the 22 influenza B Victoria lineage viruses. Notably, all four major antigenic sites (120-loop, 150-loop, 160-helix, and 190-helix regions) in the HA protein exhibited variations. Although no mutations were detected at resistance sites on the NA protein, a change occurred in the glycosylation site at position 197 NETQ in the HA protein.Conclusions:The main amino acid sites of the HA protein of the influenza B Victoria lineage viruses in Tongling City from 2019 to 2022 have undergone significant variation, which may lead to antigenic drift. Therefore, it is essential to strengthen the monitoring of influenza virus mutations.

Objective To investigate the significance of plasma levels of insulin-like growth factor(IGF)-1 and insulin-like growth factor binding protein(IGFBP)-3 in predicting acute respiratory distress syndrome(ARDS)and prognosis in critical patients.Methods A totally of 131 critical patients in intensive care unit(ICU)of the First Affiliated Hospital of Wenzhou Medical University were reviewed.Plasma concentrations of IGF-1,IGFBP-3,blood biochemistry,procalcitonin(PCT),lactic acid(LAC)and blood albumin were measured in enrolled patients.The 60-day fatality of enrolled patients was calculated.The differences between ARDS group and control group,as well as 60-day dead group and survival group were compared.Results Plasma IGF-1 and IGFBP-3 in ARDS group were significantly lower than those in control group,while plasma PCT was higher than that in control group.Plasma levels of IGF-1 and IGFBP-3 in dead group were significantly lower than those in survival group.Multivariate Logistic regression analysis and receiver operating characteristic curve results showed that IGF-1 area under curve(AUC)was 0.770,sequential organ failure assessment(SOFA)(AUC=0.692)and PCT(AUC=0.710)were independent risk factors for ARDS in critical patients.IGF-1(AUC=0.807),IGFBP-3(AUC=0.759)and SOFA score(AUC=0.859)were independent risk factors for death in critical patients.Conclusion The plasma levels of IGF-1 and IGFBP-3 in critica patients are significantly decreased,which may be an important factor for ARDS risk and fatality in critical patients.

Objective:To investigate effect of curcumin on sepsis-induced acute lung injury rats based on Nrf2/Keap1 signaling pathway.Methods:Thirty rats were randomly divided into sham operation group,model group,curcumin low,medium and high doses groups,with 6 rats in each group.Rats in sham operation group were exposed by removing cecum and then returned to abdominal cavity,while rest of rats were constructed a model of acute lung injury in sepsis,and drugs were administered immediately after surgery for 3 consecutive days.After anaesthesia with 40 mg/kg pentobarbital,blood and alveolar lavage fluid were collected from common carotid arteries,and lung tissues were taken after death.Severity of sepsis in rats was evaluated;total cell count and neutrophil count of alveo-lar lavage fluid were detected;HE staining was performed to observe pathological damage of lung tissues;colorimetric method was used to detect SOD and MDA activities of lung tissues;ELISA was used to detect TNF-α,IL-1β and IL-6 levels;qRT-PCR was used to detect Nrf2,Keap1 and HO-1 mRNA expressions in lung tissues;Western blot was used to detect Nrf2,Keap1 and HO-1 protein expressions in lung tissues.Results:Compared with sham operation group,severity scores of sepsis in model group,curcumin low,medium and high doses groups were increased,total number of cells and neutrophil number were increased,SOD level was decreased,MDA level was increased,TNF-α,IL-1β and IL-6 levels were increased,Nrf2,HO-1 mRNA and protein expressions were decreased,Keap1 mRNA and protein expressions were increased(P<0.05);compared with model group,sepsis severity scores of curcumin low,medium and high doses groups were decreased,total number of cells and neutrophil number were decreased,SOD level was increased,MDA level was decreased,TNF-α,IL-1β and IL-6 levels were decreased,Nrf2 and HO-1 mRNA and protein expressions were increased,Keap1 mRNA and protein expressions were decreased in a dose-dependent manner(P<0.05).Conclu-sion:Curcumin can improve acute lung injury in rats with sepsis,improve lung pathological injury and protect lung tissue,which may be achieved by regulating Nrf2/Keap1 pathway and expressions of related factors.

Objective To investigate the effects of long-term exposure to three new types of light emitting diode (LED) sources on the behavior, learning, and memory of rats. Methods A total of 160 specific pathogen-free SD rats were divided into eight groups as followed, trichromatic fluorescent lamps color temperature control group, violet-chip full-spectrum white LED group, blue-chip white LED group, and blue-chip full-spectrum white LED group based on the light sources types, with color temperature of 4 000 K and 6 500 K groups in each group using the 4×2 factorial design. There were 20 rats in each group, with half of the rats were males and half females. Rats were exposed to artificial lighting, and the illumination was set at 750 lx. The rats in each group were exposed to different lighting environments for 12 hours per day for 24 weeks. The open-field and step-down tests were conducted in rats after 24 weeks exposure, followed by sacrifice of rats and measurement of organ coefficients. Differences in body weight, organ coefficients, and neurobehavioral indexes of rats in different groups were compared. Results The spleen coefficient of female rats decreased in blue-chip white LED of 6 500 K color temperature group, and the liver coefficient of male rats decreased in the violet-chip full-spectrum white LED of 4 000 K color temperature, blue-chip full-spectrum white LED of 4 000 K color temperature, and blue-chip full-spectrum white LED of 6 500 K color temperature groups, compared with the same-sex rats in trichromatic fluorescent lamps with same-color temperature control group (all P<0.05). The result of different types of light sources compared in the open-field test showed that the index of total distance and movement speed of female rats in the blue-chip full-spectrum white LED group were lower than those in the other three groups, and the time cost to the central area was longer than that in the blue-chip white LED group and the violet-chip full-spectrum white LED group (all P<0.05). The total distance and movement speed of male rats in the blue-chip full-spectrum white LED group were longer or higher than those in the violet-chip full-spectrum white LED group (all P<0.05). Based on the comparison of color temperature, the time and total distance of male rats in 6 500 K color temperature group were lower than that in the 4 000 K color temperature group (both P<0.05). In the step-down test, both male and female rats in the blue-chip full-spectrum white LED group made more errors compared with other three groups with the same gender (all P<0.05). Conclusion Based on the experimental conditions of this study, the blue-chip full-spectrum white light LED affects behavior, learning and memory of the rats, and trichromatic fluorescent lamp has the lowest effect on neurobehavior. The color temperature also affects behavior of the rats, and high color temperature has higher risk.

ObjectiveTo investigate the prevalence of overweight or obesity in community patients with schizophrenia in Shanghai and to explore the related factors. MethodsStratified cluster sampling method was used and the general condition， physical examination and laboratory examination data of patients with schizophrenia who voluntarily participated in 2020 free health examination of National Basic Public Health Service were analyzed. ResultsA total of 3 200 patients were included into the study ，and the prevalence of overweight and obesity was 36.75% and 17.19%， respectively. Multivariate logistic regression analysis indicated that age between 40 and 60 （OR=1.333， 95%CI： 1.030‒1.724）， intake of first-generation antipsychotics （OR=1.413， 95%CI： 1.112‒1.796）， intake of second-generation antipsychotics （OR=1.573， 95%CI： 1.288‒1.921）， high-normal blood pressure （OR=1.549， 95%CI： 1.245‒1.927）， high-abnormal blood pressure （OR=2.824， 95%CI： 2.204‒3.619）， elevated ALT （OR=1.874， 95%CI： 1.386‒2.535）， elevated FBG （OR=1.270， 95%CI： 1.066‒1.513）， and elevated TG （OR=1.652， 95%CI： 1.335‒2.044） were the related factors that associated overweight or obesity in patients with schizophrenia. ConclusionOverweight and obesity are highly prevalent among community patients with schizophrenia in Shanghai. Age between 40 and 60， taking first-generation and second-generation antipsychotics， blood pressure higher than 120/80 mmHg， elevated ALT， elevated FBG， and elevated TG are associated with overweight or obesity in patients with schizophrenia. To provide personalized health guidance， medical staff in primary health care institutions should pay more attention to high-risk groups of overweight and obesity in schizophrenia patients at annual physical examination.

Abstract: Objective　To investigate the early diagnostic value of peripheral blood procalcitonin (PCT), C-reactive protein (CRP), fibrinogen (FIB) and D-dimer (D-D) levels in patients with pulmonary tuberculosis (PTB) complicated with bacterial pneumonia. Methods　A total of 102 patients who admitted to Department of Tuberculosis of Affiliated Nantong Hospital of Shanghai University from Jan 2021 to May 2022 were enrolled in this study and divided into a group (52 cases) with pulmonary tuberculosis (PTB) patients and a group (50 cases) with PTB patients complicated with bacterial pneumonia. The levels of PCT, CRP, FIB and D-D in the peripheral blood were measured, the differences and correlations in all indicators were compared among two groups. The sensitivity and specificity of these indicators in the early diagnosis of PTB complicated with bacterial pneumonia were analyzed by receiver operating characteristic (ROC) curve. Results　The levels of PCT, CRP, FIB and D-D in the peripheral blood from the PTB complicated with bacterial pneumonia group were 0.06 (0.04, 0.16) ng/mL, 38.00 (3.88, 96.10) mg/L, 4.51 (3.02, 6.07) g/L, and 0.59 (0.34, 1.88) mg/L, respectively, which were significantly higher than corresponding 0.04 (0.03, 0.04) ng/mL, 3.20 (0.84, 7.22) mg/L, 2.96 (2.48, 3.77) g/L, and 0.27 (0.17, 0.36) mg/L in the PTB group (Z=-4.784, -5.233, -3.853, -4.199, all P<0.001). Furthermore, the levels of CRP and FIB in the PTB complicated by bacterial pneumonia group were highly positively correlated (r=0.855, P<0.001). The area under the ROC curve (AUC) of PCT, CRP, FIB and D-D for early diagnosis of PTB complicated with bacterial pneumonia were 0.757, 0.794, 0.747 and 0.764, respectively. In addition, the AUC obtained by simultaneous measurement of PCT, CRP, FIB and D-D was as high as 0.916, and the sensitivity and specificity of diagnosing PTB complicated with bacterial pneumonia were increased to 85.7% and 96.9%, respectively, which were higher than those of individual indicators. Conclusions　Levels of peripheral blood PCT, CRP, FIB, and D-D all show varying degrees of increase in patients with PTB complicated with bacterial pneumonia, and detecting the levels of all four markers, rather than any single marker, can assist in early monitoring whether the tuberculosis patients are complicated with bacterial pneumonia.

Objective:To explore the effect of signal transducer and activator of transcription 6 (STAT6) on ferroptosis in skeletal muscle cells in sepsis model and its potential mechanism.Methods:Twenty-four 8-week-old male specific pathogen free Kunming mice were divided into normal control group, sham group, sepsis model group and STAT6 inhibitor pretreatment group according to random number table method with 6 mice in each group. A mouse sepsis model was reproduced by cecal ligation and perforation (CLP). In the sham group, the skin of mice was sutured after exposing the cecum tissue. In the STAT6 inhibitor pretreatment group, 10 mg/kg AS1517499 was injected intraperitoneally 1 hour before model reproduction. The sham group and the model group were intraperitoneally injected with the same volume of normal saline. Mice in the normal control group did not receive any operation or drug intervention. The mice were sacrificed 24 hours after model reproduction, and the muscle tissue of hind limb was obtained under sterile condition. Hematoxylin-eosin (HE) staining was used to observe the histopathology with optical microscope, and mitochondrial morphological changes were observed by transmission electron microscopy after double staining with uranium acetate lead citrate. The ferroptosis marker proteins expressions of chitinase-3-like protein 1 (CHI3L1), cyclooxygenase-2 (COX-2), acyl-CoA synthetase long-chain family member 4 (ACSL4), ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPx4) were detected by Western blotting.Results:Under the optical microscope, the morphology and structure of skeletal muscle tissues in the normal control and sham groups were normal. In the model group, the structure of skeletal muscle tissues was loose, the muscle fiber became smaller and atrophic, inflammatory cell infiltration and even muscle fiber loss were found. Compared with the model group, the structure of skeletal muscle tissues was tight and skeletal muscle atrophy was improved in the STAT6 inhibitor pretreatment group. The ultrastructure of skeletal muscle cell in the normal control and sham groups was normal under transmission electron microscope. The ultrastructure characteristics of skeletal muscle in the model group showed that cell membrane was broken and blister, mitochondria became smaller and membrane density increased, the mitochondrial crista decreased or disappeared, the mitochondrial outer membrane was broken, and the nucleus was normal in size but lacked chromatin condensation. Compared with the model group, the STAT6 inhibitor pretreatment group had a significant improvement in the ultrastructure of muscle cells. Compared with the normal control and sham groups, the protein expressions of CHI3L1, COX-2, ACSL4 and FTH1 in the muscle of the model group were significantly increased, while the protein expression of GPx4 was decreased significantly, indicating that the skeletal muscle cells in the mouse sepsis model showed characteristic mitochondrial injury and abnormal expression of ferroptosis markers. Compared with the model group, the protein expressions of CHI3LI, COX-2, ACSL4 and FTH1 in the STAT6 inhibitor pretreatment group were significantly decreased [CHI3L1 protein (CHI3L1/GAPDH): 0.70±0.08 vs. 0.97±0.09, COX-2 protein (COX-2/GAPDH): 0.61±0.03 vs. 0.83±0.03, ACSL4 protein (ACSL4/GAPDH): 0.75±0.04 vs. 1.02±0.16, FTH1 protein (FTH1/GAPDH): 0.49±0.06 vs. 0.76±0.13, all P < 0.05], while the protein expression of GPx4 was significantly increased (GPx4/GAPDH: 1.14±0.29 vs. 0.53±0.03, P < 0.05). Conclusions:Sepsis can induce ferroptosis in skeletal muscle cells of mice. STAT6 may mediate ferroptosis in mouse skeletal muscle cells by regulating the expressions of COX-2, ACSL4, FTH1 and GPx4, thereby inducing skeletal muscle cell injury in sepsis.