Cryopreservation is the primary technique for in vitro preservation of allogeneic tissue. However, its success is often hindered by factors such as low temperature, ischemia, and hypoxia. This study investigated the potential of Sini Decoction, known for its antioxidant and anti-apoptotic properties, to reduce cryopreservation-induced injury in rats' sciatic nerves. Sini Decoction was prepared according to the Chinese Pharmacopoeia, and its cytotoxicity on Rsc96 cells was assessed by using the CCK-8 method. Sini Decoction at concentrations of 4, 8, and 16 mg·mL~(-1), termed as low-(SL), medium-(SM), and high-(SH) doses group, was used for cryopreservation of rats' sciatic nerves. A normal control(NC) group and a fresh nerve control(fresh) group were set. Flow cytometry and TUNEL staining were used to detect the apoptosis of neural tissue cells after cryopreservation. Western blot was used to detect the expression of apoptosis-related proteins(Bcl-2, Bax, caspase-3, and caspase-8) and nerve regeneration proteins(NGF and BDNF) in vitro after cryopreservation. Oxidative damage of neural tissue after cryopreservation was evaluated by measuring levels of GSH, SOD, MDA, ROS, and ATP. Cryopreserved nerves were then used for allogeneic transplantation. One week after transplantation, CD4~+ and CD8~+ fluorescent double staining assessed inflammatory cell invasion in the transplanted nerve segment, and ELISA evaluated the expression of serum inflammatory factors(IL-1, IFN-γ, and TNF-α) in recipients. Twenty weeks after transplantation, electrophysiology and NF200 neurofilament staining were used to evaluate nerve regeneration. RESULTS:: showed that Sini Decoction at concentrations of below 32 mg·mL~(-1) exhibited no cytotoxicity to Rsc96 cells. During in vitro nerve cryopreservation, Sini Decoction significantly reduced cell apoptosis, ROS, and MDA production compared to the NC group. In the SH group, the protein expression of NGF and BDNF in vitro, as well as ATP, SOD, and GSH production, were significantly increased. In the rejection reaction one week after transplantation, compared to the fresh nerve transplantation group, the SL and SM groups showed reduced CD4~+ and CD8~+ T cell invasion in the transplanted nerve segment and down-regulated IL-1, IFN-γ, and TNF-α expression in recipient serum. Twenty weeks after transplantation, the electrophysiological test results of CMAP, NCV, and NF200 neurofilament protein fluorescent staining in the SM and SH groups were superior to those in the NC and fresh groups. These findings indicate that Sini Decoction offers protective benefits in the cryopreservation of rats' sciatic nerves and holds significant potential for the in vitro preservation of tissue and organs.
Animals ; Apoptosis/drug effects* ; Rats ; Oxidative Stress/drug effects* ; Sciatic Nerve/cytology* ; Cryopreservation ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Rats, Sprague-Dawley ; Protective Agents/pharmacology*

This study aims to explore the impact of host plants on the quality of Taxilli Herba and provide a theoretical basis for the quality control of Taxilli Herba. The components of Taxilli Herba from three different host plants(Morus alba, Salix babylonica, and Cinnamomum cassia) and its 3 hosts(mulberry branch, willow branch, and cinnamon branch) were detected by widely targeted metabolomics based on ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Principal component analysis(PCA), orthogonal partial least squares discriminant analysis(OPLS-DA), and Venn diagram were employed for analysis. A total of 717 metabolites were detected in Taxilli Herba from the three host plants and the branches of these host plants by UPLC-MS/MS. The results of PCA and OPLS-DA of Taxilli Herba from the three different host plants showed an obvious separation trend due to the different effects of host plants. The Venn diagram showed that there were 32, 8, and 26 characteristic metabolites in samples of Taxilli Herba from M. alba host, S. babylonica host, and C. cassia host, respectively. It was found by comparing the characteristic metabolites of Taxilli Herba and its hosts that each host transmits its characteristic components to Taxilli Herba, so that the Taxilli Herba contains the characteristic components of the host. The Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis showed that the differential metabolites of Taxilli Herba from the three hosts were mainly enriched in flavonoid biosynthesis, arginine and proline metabolism, and glycolysis/gluconeogenesis pathways. Furthermore, the differential metabolites enriching pathways of Taxilli Herba from the three hosts were different depending on the host. In a word, host plants have a significant impact on the metabolites of Taxilli Herba, and it may be an important factor for the quality of Taxilli Herba.
Metabolomics/methods* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid ; Tandem Mass Spectrometry ; Quality Control ; Salix/chemistry* ; Cinnamomum aromaticum/metabolism* ; Principal Component Analysis

OBJECTIVE: To evaluate the protective effects of resveratrol against acute lung injury (ALI) and investigate the potential mechanisms underlying the reactive oxygen species (ROS)-triggered thioredoxin-interacting protein (TXNIP)/NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) pathway. METHODS: C57BL/6 mice and J774A.1 cells were selected as the research subjects. Thirty Mice were randomly divided into 5 groups of 6 in each group: control with 0.9% saline, 5 mg/kg lipopolysaccharide (LPS) 24 h, 25 mg/kg resveratrol + 5 mg/kg LPS, 100 mg/kg resveratrol + 5 mg/kg LPS, and 4 mg/kg NLRP3 inhibitor CY-09 + 5 mg/kg LPS. For cell stimulation, cells were pretreated with 5 and 20 µmol/L resveratrol for 2 h, and stimulated with or without 1 µg/mL LPS and 3 mmol/L ATP for 2 h. The antioxidant N-acetyl-L-cysteine (NAC, 2 µmol/L) was used as the positive control group. Hematoxylin and eosin staining was used to evaluate the degree of lung LPS-induced tissue damage, and enzyme-linked immunosorbent assay was used to evaluate the contents of interleukin-1 β (IL-1 β) and IL-18 in the serum and cell supernatant. ROS and malondialdehyde (MDA) levels in the lung tissue were detected using the corresponding kits. Western blotting was used to detect the expressions of TXNIP, high-mobility group box 1 (HMGB1), NLRP3, as well as cysteine-aspartic acid protease 1 (caspase-1) and gasdermin D (GSDMD) along with their cleaved forms in lung tissue. Additionally, reverse transcription quantitative polymerase chain reaction was performed to analyze the expression of related inflammatory cytokines. ROS content was detected using flow cytometry and confocal laser microscopy. Mitochondrial morphological changes were observed using transmission electron microscopy, and HMGB1 expression was detected using immunofluorescence. RESULTS: Resveratrol significantly alleviated LPS-induced lung damage with reduced inflammation, interstitial edema, and leukocyte infiltration (P<0.01). It also decreased serum levels of IL-1 β and IL-18 (P<0.05), while downregulating the expressions of NLRP3, IL-6, and other inflammatory markers at both the protein and mRNA levels (P<0.05). Notably, the higher dose (100 mg/kg) demonstrated a better effect than the lower dose (25 mg/kg). In macrophages, resveratrol reduced IL-1 β and IL-18 following LPS and ATP stimulation, suppressed HMGB1 translocation, and inhibited formation and activation of the NLRP3 inflammasome (P<0.05 or P<0.01). These anti-inflammatory effects were mediated through the suppression ROS accumulation (P<0.01) and mitochondrial dysfunction. Transmission electron microscopy revealed that resveratrol preserved mitochondrial structure, preventing the mitochondrial damage seen in LPS-treated groups (P<0.01). The expressions of cleaved caspase-1, cleaved GSDMD, and cytoplasmic HMGB1 were all reduced following resveratrol treatment (P<0.01). Moreover, resveratrol inhibited dissociation of TXNIP from thioredoxin, blocking subsequent activation of NLRP3 and downstream inflammatory cytokines (P<0.01). Similarly, the higher concentration of resveratrol (20 µ mol/L) exhibited superior efficacy in vitro. CONCLUSION Resveratrol can reduce the inflammatory response following ALI and inhibit the activation of NLRP3 inflammasome and the level of HMGB1 in the cytoplasm by inhibiting ROS overproduction.
Acute Lung Injury/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Animals ; Resveratrol/pharmacology* ; Reactive Oxygen Species/metabolism* ; Inflammation/complications* ; Mice, Inbred C57BL ; Carrier Proteins/metabolism* ; Signal Transduction/drug effects* ; Lipopolysaccharides ; Thioredoxins/metabolism* ; Mice ; Lung/drug effects* ; Male ; Cell Line ; Interleukin-1beta/metabolism* ; Cell Cycle Proteins ; Stilbenes/therapeutic use*

The angiogenic response is essential for the repair of ischemic brain tissue. Integrin α6 (Itga6) expression has been shown to increase under hypoxic conditions and is expressed exclusively in vascular structures; however, its role in post-ischemic angiogenesis remains poorly understood. In this study, we demonstrate that mice with endothelial cell-specific knockout of Itga6 exhibit reduced neovascularization, reduced pericyte coverage on microvessels, and accelerated breakdown of microvascular integrity in the peri-infarct area. In vitro, endothelial cells with ITGA6 knockdown display reduced proliferation, migration, and tube-formation. Mechanistically, we demonstrated that ITGA6 regulates post-stroke angiogenesis through the PI3K/Akt-eNOS-VEGFA axis. Importantly, the specific overexpression of Itga6 in endothelial cells significantly enhanced neovascularization and enhanced the integrity of microvessels, leading to improved functional recovery. Our results suggest that endothelial cell Itga6 plays a crucial role in key steps of post-stroke angiogenesis, and may represent a promising therapeutic target for promoting recovery after stroke.
Animals ; Nitric Oxide Synthase Type III/metabolism* ; Mice ; Proto-Oncogene Proteins c-akt/metabolism* ; Integrin alpha6/genetics* ; Endothelial Cells/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Stroke/pathology* ; Vascular Remodeling/physiology* ; Vascular Endothelial Growth Factor A/metabolism* ; Mice, Knockout ; Signal Transduction/physiology* ; Mice, Inbred C57BL ; Male ; Neovascularization, Physiologic/physiology*

Objective:To understand the tobacco use situation and related factors among the occupational population in Beijing in 2018.Methods:The data were obtained from a special survey on “Healthy Beijing People—Ten-year Action Plan for Health Promotion (2009—2018)”. From June to September 2018, 12 908 participants were recruited from 16 districts of Beijing using the stratified sampling method combined with the typical sampling method. Questionnaires were conducted to collect basic information, work situations, tobacco exposure in public places, and smoking cessation. The Chi-square test was used to compare the differences in current smoking rates, smoking cessation rates and second-hand smoke exposure rates among participants with different characteristics. Multivariate logistic regression analysis was used to analyze the related factors of tobacco use.Results:The present smoking rate of the occupational population in Beijing was 18.0% (95% CI: 17.3%-18.7%), and the passive smoking exposure rate of non-smokers was 61.0% (95% CI: 60.1%-61.9%). The smoking cessation rate among the working population was 22.2% (95% CI: 20.7%-23.7%), and the proportion of current smokers with the intention to quit was 63.9% (1 485/2 325). The present smoking rate of employees of different genders, ages, education levels and occupational nature showed statistically significant differences ( P<0.05). The results of multivariate logistic regression analysis showed that in males, compared with the age group of 16-29 years old, the risk of smoking increased in the age groups of 30-39 years old, 40-49 years old and over 50 years old [ OR (95% CI) values were 1.49 (1.27-1.74), 1.34 (1.13-1.59) and 1.30 (1.06-1.59)]. Compared with those with junior high school or lower education, participants with high school/vocational/college education group had a higher risk of smoking ( OR=1.38, 95% CI: 1.13-1.69), while those with master′s degree or above had a lower risk of smoking ( OR=0.30, 95% CI: 0.22-0.42). The physical workers ( OR=1.28, 95% CI: 1.11-1.47) had a higher risk of smoking than the mental workers. Conclusion:The current smoking rate among the occupational population in Beijing is lower than the national level during the same period, but the exposure rate to second-hand smoke is relatively high. Age, educational level, job nature and gender are related factors of smoking.

Angiogenesis is a key link in the development of atherosclerotic plaques.Inhibiting angiogenesis con-tributes to plaque stabilization and reduces the risk of related cardiovascular events.Apolipoprotein A1 binding protein(A1 BP),an important secretory protein,has been shown in a growing body of research to play a significant role in the reg-ulation of angiogenesis.This article aims to elucidate the mechanisms of action of A1 BP on angiogenesis and cardiovascu-lar diseases,thereby providing new perspectives for the clinical treatment of cardiovascular diseases.

Objective:To understand the tobacco use situation and related factors among the occupational population in Beijing in 2018.Methods:The data were obtained from a special survey on “Healthy Beijing People—Ten-year Action Plan for Health Promotion (2009—2018)”. From June to September 2018, 12 908 participants were recruited from 16 districts of Beijing using the stratified sampling method combined with the typical sampling method. Questionnaires were conducted to collect basic information, work situations, tobacco exposure in public places, and smoking cessation. The Chi-square test was used to compare the differences in current smoking rates, smoking cessation rates and second-hand smoke exposure rates among participants with different characteristics. Multivariate logistic regression analysis was used to analyze the related factors of tobacco use.Results:The present smoking rate of the occupational population in Beijing was 18.0% (95% CI: 17.3%-18.7%), and the passive smoking exposure rate of non-smokers was 61.0% (95% CI: 60.1%-61.9%). The smoking cessation rate among the working population was 22.2% (95% CI: 20.7%-23.7%), and the proportion of current smokers with the intention to quit was 63.9% (1 485/2 325). The present smoking rate of employees of different genders, ages, education levels and occupational nature showed statistically significant differences ( P<0.05). The results of multivariate logistic regression analysis showed that in males, compared with the age group of 16-29 years old, the risk of smoking increased in the age groups of 30-39 years old, 40-49 years old and over 50 years old [ OR (95% CI) values were 1.49 (1.27-1.74), 1.34 (1.13-1.59) and 1.30 (1.06-1.59)]. Compared with those with junior high school or lower education, participants with high school/vocational/college education group had a higher risk of smoking ( OR=1.38, 95% CI: 1.13-1.69), while those with master′s degree or above had a lower risk of smoking ( OR=0.30, 95% CI: 0.22-0.42). The physical workers ( OR=1.28, 95% CI: 1.11-1.47) had a higher risk of smoking than the mental workers. Conclusion:The current smoking rate among the occupational population in Beijing is lower than the national level during the same period, but the exposure rate to second-hand smoke is relatively high. Age, educational level, job nature and gender are related factors of smoking.

Angiogenesis is a key link in the development of atherosclerotic plaques.Inhibiting angiogenesis con-tributes to plaque stabilization and reduces the risk of related cardiovascular events.Apolipoprotein A1 binding protein(A1 BP),an important secretory protein,has been shown in a growing body of research to play a significant role in the reg-ulation of angiogenesis.This article aims to elucidate the mechanisms of action of A1 BP on angiogenesis and cardiovascu-lar diseases,thereby providing new perspectives for the clinical treatment of cardiovascular diseases.

Objective:To investigate the clinical effect of Alirocumab in reducing low density lipoprotein cholesterol(LDL-C)in patients with coronary heart disease(CHD)and substandard blood lipids and its effect on atheroscle-rotic plaque.Methods:A total of 127 CHD patients with substandard blood lipids who were treated in Shenzhen Longhua District People's Hospital between September 2017 and March 2021 were selected.According to patients re-ceived Alirocumab therapy or not,they were divided into control group(n=81)and combined group(n=46).The control group remained original statin therapeutic regimen,while combined group received additional PCSK9 inhibi-tor Alirocumab injection based on original statin therapeutic regimen,75mg subcutaneously,once every 2 weeks.Both groups were treated for 3 months,then followed up for 12 months.Levels of total cholesterol(TC)and LDL-C,plaque fiber cap thickness,lipid plaque radian,lipid plaque length and minimum lumen cross-sectional area before and after 12-month follow-up,and incidence of adverse reactions and clinical endpoint events were com-pared between two groups.Results:After 12-month follow-up,compared with control group,combined group had significant lower plasma levels of TC[(3.48±1.04)mmol/L vs.(2.29±0.76)mmol/L],LDL-C[(2.08±0.53)mmol/L vs.(1.27±0.41)mmol/L],lipid plaque radian[(107.22±13.29)° vs.(92.65±11.81)°]and lipid plaque length[(5.45±0.89)mm vs.(4.84±0.82)mm],and significant higher plaque fiber cap thickness[(123.60±14.87)μm vs.(131.46±14.29)μm]and minimum lumen cross-sectional area[(2.51±0.37)mm2 vs.(2.69±0.33)mm2](P＜0.01 all).There was no significant difference in incidence rates of adverse reactions(x2=0.428,P=0.513)and clinical endpoint events(x2=0.253,P=0.615)between two groups.Conclusion:Alirocumab can significantly reduce LDL-C level,increase the plaque fiber cap thickness and lumen cross-section-al area,reduce the internal lipid load of plaque and improve the stability of plaque with good safety in CHD patients with substandard blood lipids.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.