Objective To explore the effects and mechanisms of human umbilical cord mesenchymal stem cell (HUC-MSC)-derived exosomes (Exo) pretreated with Huangkui capsules on renal ischemia-reperfusion injury (IRI). Methods HUC-MSCs were cultured in media containing different concentrations of Huangkui capsules for 24 hours to determine cell viability and select an appropriate concentration for subsequent experiments. HUC-MSCs were pretreated with 50 μg/mL Huangkui capsules for 24 hours, and Exo were extracted using an exosome extraction kit. The morphology was observed under a transmission electron microscope, particle size was measured by nanoparticle tracking analysis, and the expression of exosomal membrane surface marker proteins was detected by Western blot. Human renal tubular epithelial cells (HK-2 cells) were randomly divided into hypoxia/reoxygenation group (M group), hypoxia/reoxygenation + Exo group (E group), and hypoxia/reoxygenation + Huangkui capsules pretreated Exo group (H group). Western blotting was used to measure the expression of endoplasmic reticulum stress (ERS)-related proteins, and real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to measure the expression of ERS-related gene messenger RNA (mRNA). Mice were randomly divided into sham operation group (Sham group), ischemia-reperfusion group (I/R group), ischemia-reperfusion + Exo group (E group), and ischemia-reperfusion + Huangkui capsules pretreated Exo group (H group). Renal histological assessment, serum creatinine (Scr), blood urea nitrogen (BUN) measurement and inflammatory factor detection were performed 24 hours later. Results Both Exo and Huangkui capsules prereated Exo had a bilayer membrane structure and a cup-shaped morphology; their average particle sizes were 116.8 nm and 81.3 nm, respectively. Both expressed CD9, CD63, TSG101. Compared with the M group, the E group had decreased relative expression of transcription factor 6 (ATF6) and protein kinase R-like endoplasmic reticulum kinase (PERK) proteins, increased mRNA relative expression, increased relative expression of C/EBP homologous protein (CHOP) protein, and decreased mRNA relative expression. Compared with the E group, the H group had decreased relative expression of ATF6, PERK, CHOP proteins, and decreased mRNA relative expression of ATF6 and PERK (all P<0.05). Animal experimental results showed that compared with the Sham group, the I/R group had increased renal tubular injury scores, Scr, BUN, interleukin (IL)-1β, IL-10, IL-18, tumor necrosis factor (TNF)-α levels. Compared with the I/R group, the E and H groups had decreased renal tubular injury scores and Scr, BUN, IL-1β, IL-10, IL-18, TNF-α levels. Compared with the E group, the H group had decreased renal tubular injury scores and Scr, BUN, IL-1β, IL-10, IL-18, TNF-α levels (all P<0.05). Conclusions Huangkui capsules pretreatment HUC-MSC-derived Exo may alleviate renal IRI by inhibiting ERS.

ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription （ZJTP） on human umbilical vein endothelial cells （HUVECs） damaged by high glucose combined with lipopolysaccharide （LPS）. MethodThe survival rate of cells was determined by cell counting kit-8 （CCK-8）， and the level of tumor necrosis factor-α （TNF-α） was determined by enzyme-linked immunosorbent assay （ELISA） to determine the optimal injury concentration and action time of LPS， as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group， a model group， a ZJTP drug-containing serum group， and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 （ET-1）， nitric oxide （NO）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 （GPR43）， β-suppressor protein-2 （β-arrestin-2）， nuclear factor-κB suppressor α （IκBα）， and nuclear factor κB p65 （NF-κB p65）. The nucleation of NF-κB p65 was observed by immunofluorescence staining （IF）. The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid （siRNA）. The cells after intervention were divided into an empty carrier group， a ZJTP drug-containing serum group， a Si-GPR43 group， and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β， IL-6， and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L^-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group， the content of ET-1 in the model group was significantly increased， and the content of NO was significantly decreased （P<0.01）. The levels of inflammatory factors were significantly increased （P<0.01）. The expressions of GPR43 and IκBα were significantly decreased， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased （P<0.01）. NF-κB p65 protein was transferred from the extranuclear to the intranuclear （P<0.01）. Compared with the model group， the content of ET-1 in the ZJTP drug-containing serum group was decreased， and the content of NO was increased （P<0.05）. The levels of inflammatory factors decreased （P<0.05）. The protein expressions of GPR43 and IκBα were increased， while the expressions of β-arrestin-2 and NF-κB p65 were decreased （P<0.05）. The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased （P<0.01）. The mechanism study showed that compared with the Si-GPR43 group， the content of IL-1β， IL-6， and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum （P<0.01）. The protein expressions of GPR43 and IκBα were significantly increased （P<0.01）， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased （P<0.01）. The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased （P<0.01）. ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury， which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.

Based on the neurovascular unit(NVU), neurovascular coupling functions as a barrier to maintain the homeostasis of the microenvironment by regulating the signaling and metabolic activity of nerve cells and capillaries. Widely dispersed across the retina, the NVU is essential to preserving its normal physiological function. A disturbance in retinal neurovascular homeostasis produced by a range of factors can result in a variety of retinal disorders, such as diabetic retinopathy(DR), glaucoma, retinitis pigmentosa(RP)and age-related macular degeneration(ARMD). The retina also has a widespread distribution of brain-derived neurotrophic factor(BDNF), which functions to promote neuron growth and repair damage by binding to its receptor TrkB. In recent years, BDNF was found to play a protective role against damage in the early stage of retinal neurovascular homeostasis imbalance, often known as the neurodegenerative stage. It also helps to reduce the production of pro-angiogenic substances of neurological origin and offers a fresh approach for the early detection and treatment of associated eye disorders.

Objective:To systematically review the risk prediction models for cognitive impairment in stroke patients, aiming to provide references for clinical healthcare professionals in selecting or constructing high-quality risk assessment tools.Methods:A computerized search was conducted in PubMed, Embase, Web of Science, OVID, Cochrane Library, SinoMed, CNKI, Wanfang Database, and VIP to identify studies related to risk prediction models for cognitive impairment in stroke patients. The search was limited to articles published up to August 1, 2023. Two researchers independently screened the literature, extracted data, and assessed the risk of bias and applicability of the included studies using PROBAST.Results:A total of 26 articles were included. The applicability of the studies was generally good, but all studies had some degree of bias risk, mainly arising from unreasonable study designs, inappropriate time intervals between predictor assessment and outcome determination, insufficient sample sizes, unreasonable handling of continuous variables, omission of missing data, lack of reporting of calibration, and overfitting of the models. Meta-analysis results showed that age ( OR=0.05, 95% CI: 0.033-0.057), education level ( OR=-0.13, 95% CI: -0.171 - -0.082), history of diabetes ( OR=2.32, 95% CI: 1.867-2.881), history of hypertension ( OR=0.67, 95% CI: 0.420-0.918), and NIHSS score ( OR=0.40, 95% CI: 0.331-0.469) were factors for cognitive impairment in stroke patients. Conclusions:While various risk prediction models for cognitive impairment in stroke patients exist, they suffer from methodological flaws and high bias risks, with some commonalities and controversies in predictors. Future research should adhere to the principles of transparent reporting of individual prognosis or diagnosis of multivariate prediction models, develop localized prediction models with low bias risk and good applicability, and conduct internal and external validations to demonstrate their applicability and feasibility in clinical practice.

Objective:To observe the effect of G protein alpha inhibitory subunit (Gαi) 1 and Gαi3 on signal transduction and angiogenesis induced by Netrin-1 (NTN1) and explore the possible mechanisms.Methods:Twenty male C57BL/6J mice aged 6 to 8 weeks were randomly assigned to a control group and a diabetic group, with 10 mice in each group. Diabete group mice were induced by streptozotocin to establish diabetes model. 12 weeks after modeling, quantitative real-time polymerase chain reaction and Western blot were performed to detect the expression of Ntn1, Gαi1 and Gαi3 in diabetic retinas. Additionally, 35 male C57BL/6J mice aged 2 weeks were randomly stratified into three groups: a control group, an intravitreal injection of NTN1 group (NTN1 group), and a retinal endothelial cell-specific Gαi1/Gαi3 knockdown coupled with intravitreal NTN1 injection group (Gαi1/Gαi3 eKD+NTN1 group), with 15 mice in each of the normal control and NTN1 groups, and 5 mice in the Gαi1/Gαi3 eKD+NTN1 group. Isolectin B4 staining was performed to observe retinal neovascularization. In vitro, human umbilical vein endothelial cells (HUVEC) were divided into four groups: negative control lentiviral transfection group (shC group), negative control lentiviral transfection+NTN1 treatment group (shC+NTN1 group), Gαi1/Gαi3 knockdown group (shGαi1/Gαi3 group), and Gαi1/Gαi3 knockdown+NTN1 treatment group (shGαi1/Gαi3+NTN1 group). The effects of NTN1, Gαi1, and Gαi3 on HUVEC proliferation were assessed using the EdU assay. Transwell assays were conducted to determine the effects on HUVEC migration, and Matrigel assays were used to evaluate the effects on HUVEC tube formation. Protein kinase B (Akt), phosphorylated Akt (p-Akt), ribosomal protein S6 kinase (S6K), phosphorylated S6K (p-S6K), extracellular regulatory protein kinase (Erk1/2), phosphorylated Erk1/2 (p-Erk1/2) protein expression on HUVEC were detected by Western blot.Results:Compared with the control group, the relative expression levels of Ntn1, Gαi1, and Gαi3 mRNA and protein in the diabetic group retina were significantly increased, with statistically significant differences ( t=11.800, 9.298, 10.620, 7.503, 3.432, 8.037; P<0.000 1). Compared with the shC group, the relative expression levels of Gαi1 and Gαi3 mRNA and protein in the shGαi1/Gαi3 group in HUVEC were significantly reduced, showing statistically significant differences ( t=16.310, 16.300, 13.600, 9.068; P<0.000 1). HUVEC proliferation rate, migration number and lumen formation number: compared with shC group, shC+NTN1 group significantly increased, while shGαi1/Gαi3 group and shGαi1/Gαi3+NTN1 group significantly decreased, and the differences were statistically significant ( F=62.750, 49.830, 54.900; P<0.000 1). Compared with the control group, the relative expression levels of Gαi1 and Gαi3 mRNA and protein in retina were significantly decreased in the Gαi1/Gαi3 eKD group, showing statistically significant differences ( t=10.920, 13.460, 9.219, 10.500; P<0.000 1). Retinal neovasculogenesis area: compared with the normal control group, the area of retinal neovasculogenesis increased significantly in the NTN1 group, but decreased significantly in the Gαi1/Gαi3 eKD+NTN1 group, with statistical significance ( F=24.010, P<0.000 1). The protein expression of p-Akt relative to Akt, p-S6K relative to S6K, and p-Erk1/2 relative to Erk1/2: compared with shC group, the protein expression of shC+NTN1 group was significantly increased, while that of shGαi1/Gαi3 group and shGαi1/Gαi3+NTN1 group was significantly decreased, with statistical significance ( F=78.610, 144.400, 77.010; P<0.000 1). Conclusions:NTN1 induces Gαi1/Gαi3 to mediate activation of downstream Akt-mammalian target proteins of rapamycin and Erk1/2, thereby promoting angiogenesis in vivo and in vitro environments. Knocking down Gαi1/Gαi3 significantly reduces the NTN1-induced angiogenesis effect.

Humans ; Aortic Valve/surgery* ; Retrospective Studies ; Transcatheter Aortic Valve Replacement ; Sex Factors ; Heart Valve Prosthesis Implantation ; Treatment Outcome ; China ; Aortic Valve Stenosis/surgery* ; Risk Factors ; Risk Assessment

Humans ; Retrospective Studies ; Transcatheter Aortic Valve Replacement/adverse effects* ; Risk Assessment ; China ; Prognosis ; Aortic Valve Stenosis/surgery* ; Treatment Outcome ; Risk Factors ; Aortic Valve

Objective To investigate the effect of sepsis on vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.Methods Thirty-six adult male SPF SpragueDawley rats,aged 2-3 months,weighing 200-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sham operation group (group S) and sepsis group (group Sep).Sepsis was induced by cecum ligation and puncture (CLP) in rats anesthetized with intraperitoneal chloral hydrate 350 mg/kg.At 12 h after CLP,the sciatic nerve-pretibial muscle was prepared.Vecuronium was added to the culture medium with the final concentration of 0.08 μg/ml,and the sciatic nerve-pretibial muscle was incubated for 15 min.Before and after administration,evoked endplate potentials (EPPs) and miniature endplate potentiais (MEPPs) were recorded by using intracellular microelectrode.EPP/MEPP ratio was calculated.Results Compared to C and S groups,EPPs,MEPPs and EPP/MEPP ratio were significantly increased before and after administration in group Sep.EPPs,MEPPs and EPP/MEPP ratio were significantly lower after administration than before administration in the three groups.Conclusion Sepsis can promote acetylcholine release in neuromuscular junction,thus weakening vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.

Objective To investigate the expression of two long non-coding RNAs ( lncRNAs ) during HIV-1 infection, which were nuclear-enriched autosomal transcript 1 (NEAT1) and metastasis asso-ciated lung adenocarcinoma transcript 1 ( MALAT1 ) , and their relationships with disease progression . Methods Fifty-nine patients with HIV-1 infection and 21 healthy subjects were recruited in this study , of which 31 patients were highly active antiretroviral therapy ( HAART)-na?ve and 28 patients received HAART for more than one year with undetectable viral loads .Total RNAs were extracted from PBMC and plasma samples, respectively.The levels of NEAT1 and MALAT1 were detected by quantitative real time polymer-ase chain reaction .Results The levels of NAET1 and MALAT1 in PBMC from HAART na?ve patients were 3 to 5 times higher than those in healthy subjects (P<0.01).The levels of NAET1 and MALAT1 in PBMC from HAART treated patients were similar to those in healthy subjects .The levels of NEAT1 in plasma sam-ples from patients with HIV-1 infection were lower than those in healthy subjects , and were positively corre-lated with CD4+T cell counts (P<0.01).No significance differences in the levels of MALAT1 in plasma samples were found between those with and without HIV-1 infection (P>0.05).Conclusion This study suggested that NEAT 1 and MALAT1 might be involved in the disease progression in patients with HIV-1 in-fection.The level of NEAT1 in plasma could be used as a potential biomarker of HIV-1 infection.

BACKGROUND:The discovery and concept of pulp tissue-derived stem cells is beneficial to the understanding of tooth development and regeneration and repair mechanisms from the cellular level. OBJECTIVE:To understand the induced differentiation capacity and induced conditions in vitro of human dental pulp stem cells into neuron-like cells. METHODS:Pulp tissue was separated from human healthy third molars. Single cellsuspensions were prepared and seeded into 6-wel plates containing alpha-modified minimum essential medium supplemented with 15%fetal bovine serum. Subconfluent cultures (first passage) of colony forming cells were induced with butylhydroxy anisole, forskolin,β-mercaptoethanol, basic fibroblast growth factor. RESULTS AND CONCLUSION:Immunofluorescence and reverse transcription-PCR assay showed that human dental pulp stem cells positively expressed stro-1, Col-I, dentin sialoprotein after 2 weeks of induction. Nestin and neuron-specific enolase were strongly expressed, but the gingival fibroblasts were negatively expressed. It indicates that adult stem cells in human dental pulp have a high neuron-like celldifferentiation potential under a certain inductive condition.