ObjectiveTo investigate the protective effect of resveratrol against blood-brain barrier damage in a rat model of cerebral ischemia-reperfusion and to determine whether this effect is mediated by the C-C motif chemokine ligand 3 (CCL3)/C-C chemokine receptor type 5 (CCR5) signaling pathway　Methods　A total of 100 specific pathogen-free (SPF) male Sprague-Dawley (SD) rats, aged 6-8 weeks, were randomly divided into five groups: sham-operated (Sham), model (MCAO), resveratrol-treated (Res), resveratrol+CCR5 agonist (Res+Agonist), and resveratrol+CCR5 antagonist (Res+Antagonist) groups. The MCAO, Res, Res+Agonist, and Res+Antagonist groups were subjected to middle cerebral artery occlusion (MCAO) for 45 min followed by reperfusion to establish the rat model of cerebral ischemia, while the Sham group underwent carotid artery exposure without ischemia. After reperfusion, resveratrol (60 mg/kg) was intraperitoneally injected in Res, Res+Agonist and Res+Antagonist groups, and an equal volume of normal saline was injected in Sham and MCAO groups. Thirty minutes later, CCR5 agonist (20 μg/kg) was injected via the tail vein in the Res+Agonist group, CCR5 inhibitor (1 mg/kg) was injected via the tail vein in the Res+Antagonist group, and the equal volume of normal saline was injected via the tail vein in the Res, Sham and MCAO groups. Thereafter, resveratrol, CCR5 agonist, CCR5 inhibitor, or saline were administered every 24 hours for 3 consecutive days. After modeling, cerebral infarct volume was evaluated by 2,3,5‑triphenyltetrazolium chloride (TTC) staining. Blood‑brain barrier (BBB) permeability was assessed by Evans blue extravasation. Serum levels of the pro‑inflammatory cytokines interleukin‑1β (IL‑1β), tumor necrosis factor‑α (TNF‑α), and CCL3 were measured by enzyme‑linked immunosorbent assay (ELISA). Histopathological changes in brain tissue were observed by hematoxylin and eosin (HE) staining and Nissl staining. Immunofluorescence staining was used to detect the fluorescence intensity of the microglial marker ionized calcium‑binding adapter molecule 1 (Iba‑1) and CCR5. Western blotting analysis was used to detect proteins associated with the CCL3/CCR5 signaling pathway (CCL3, CCR5) in brain tissue, serum pro-inflammatory cytokines [phosphorylated nuclear factor-kappa B (p-NF-κB)] and blood-brain barrier-related proteins [matrix metalloproteinase 9 (MMP9) and occludin]. ResultsCompared with the Sham group, the MCAO group exhibited significantly increased levels of all injury parameters, including cerebral infarct volume, Evans blue extravasation rate in brain tissue, serum levels of the pro‑inflammatory cytokines (IL-1β, TNF-α, and CCL3), the rate of Iba-1- and CCR5-positive immunofluorescence signals in brain tissue, and the protein expression levels of CCL3, CCR5, p-NF-κB, and MMP9 (all P<0.001), accompanied by markedly decreased expression of occludin (P<0.001). HE and Nissl staining revealed pronounced histopathological damage in the MCAO group.Compared with the MCAO group, the Res group showed significant reductions in all the aforementioned injury indicators and a significant increase in occludin expression (P<0.01), together with ameliorated histopathological changes. Compared with the Res group, the Res+Agonist group displayed significantly elevated injury indicators and decreased occludin expression (P<0.05), with aggravated brain tissue damage. In contrast, the Res+Antagonist group exhibited significant decreases in injury indicators and a significant increase in occludin expression (P<0.05), along with further alleviation of histopathological injury. ConclusionResveratrol attenuates peripheral inflammation in cerebral ischemia‑reperfusion injury by targeting the CCL3/CCR5 signaling pathway, thereby exerting a protective effect on the blood‑brain barrier.

BACKGROUND:Endometrial receptivity is a key factor in embryo implantation in northwest Tibetan cashmere goats,and the expression of genes related to endometrial receptivity and their variable splicing are still unclear. OBJECTIVE:To analyze and explore genes and variable splicing events related to endometrial receptivity in northwest Tibetan cashmere goats. METHODS:On days 5 and 15 of pregnancy(representing pre receptive endometrium group and receptive endometrium group),three northwest Tibetan cashmere goats were randomly selected.Endometrial tissue was collected and stained with hematoxylin and eosin to observe tissue morphology.Immunohistochemical staining was used to detect the expression of endometrial receptive marker proteins leukemia inhibitory factor and vascular endothelial growth factor.After the total RNA was extracted and the quality test was qualified,transcriptome sequencing was performed to search differentially expressed mRNAs,lncRNAs,circRNAs,and miRNAs,perform functional prediction,and analyze alternative splicing mRNAs and lncRNAs related to endometrial receptivity. RESULTS AND CONCLUSION:(1)Compared with the pre receptive endometrium group,the expression levels of leukemia inhibitory factor and vascular endothelial growth factor proteins in the endometrial tissue of the receptive endometrium group were significantly increased.(2)The sequencing results showed that the differentially expressed genes were mostly mRNA and lncRNA genes,including 250 upregulated mRNAs,193 upregulated lncRNAs,135 downregulated mRNAs,and 123 downregulated lncRNAs,which were significantly enriched in the Wnt,Hedgehog,and Hippo signaling pathways.(3)Alternative splicing event analysis uncovered 8 differentially expressed variable splicing transcripts,which were all mRNA transcripts,including 2 downregulated and 6 upregulated,and were significantly associated with vascular endothelial growth factor receptor signaling,cell motility,and embryonic development.

Kupffer cells (KCs), as residents and sentinels of the liver, are involved in the formation of hepatic fibrosis (HF). However, the biological functions of circular RNAs (circRNAs) in KCs to HF have not been determined. In this study, the expression levels of circRNAs, microRNAs, and messenger RNAs (mRNAs) in KCs from a mouse model of HF mice were investigated using microarray and circRNA-Seq analyses. circDcbld2 was identified as a candidate circRNA in HF, as evidenced by its up-regulation in KCs. Silver staining and mass spectrometry showed that Wtap and Igf2bp2 bind to cirDcbld2. The suppression of circDcbld2 expression decreased the KC inflammatory response and oxidative stress and inhibited hepatic stellate cell (HSCs) activation, attenuating mouse liver fibrogenesis. Mechanistically, Wtap mediated the N ⁶-methyladenosine (m6A) methylation of circDcbld2, and Igf2bp2 recognized m6A-modified circDcbld2 and increased its stability. circDcbld2 contributes to the occurrence of HF by binding miR-144-3p/Et-1 to regulate the inflammatory response and oxidative stress. These findings indicate that circDcbld2 functions via the m6A/circDcbld2/miR-144-3p/Et-1 axis and may act as a potential biomarker for HF treatment.

Objective To investigate the application of cardiovascular magnetic resonance feature tracking(CMR-FT)in assessing myocardial strain in dilated cardiomyopathy(DCM)patients residing at high altitudes.Methods We retrospectively enrolled 29 DCM patients living at high altitudes(DCM-H),27 DCM patients living in a low-altitude plain environment(DCM-P),23 healthy volunteers living at a high altitude(HV-H),and 24 healthy volunteers living in a low-altitude plain environment(HV-P).All subjects underwent cine MRI scanning using a 3.0T rapid steady-state free precession sequence.The CMR images thus acquired were analyzed using cvi42,a post-processing software,to obtain left ventricular function and myocardial strain parameters.Results Compared with the HV-H group,the DCM-H group showed higher left ventricle end-diastolic volume(LVEDV)and left ventricle end-systolic volume(LVESV),and lower left ventricular ejection fraction(LVEF)and stroke volume(LVSV)(all P＜0.01).No significant difference was observed in cardiac function between the DCM-H and DCM-P groups(all P＞0.05).The absolute values of global radial strain(GRS),global circumferential strain(GCS),and global longitudinal strain(GLS)in the DCM-H group were lower than those in the HV-P group([14.5±6.5]％vs.[34.2±10.7]％,[-11.1±4.4]％vs.[-19.9±2.8]％,and[-7.7±3.2]％vs.[-13.6±4.1]％,respectively),with the differences being statistically significant(all P＜0.001).The DCM-H group had higher absolute GRS,GCS,and GCS values than the DCM-P group did([14.5±6.5]％vs.[7.0±2.7]％,[-11.1±4.4]％vs.[—5.4±2.2]％,and[—7.7±3.2]％vs.[—4.3±1.7]％,respectivley,all P＜0.01).Conclusion Myocardial strain in DCM patients living at a high altitude is lower than that in healthy volunteers living at a high altitude,but higher than that in DCM patients living in a low-altitude plain environment.CMR-FT can be used to quantitatively assess myocardial contractility in DCM patients living at a high altitude,showing promise for clinical application.

Objective:To explore clinical and genetic features of persistent asymptomatic microscopic hematuria in children.Methods:A retrospective case analysis of 135 individuals admitted to Xi ′an Children′s Hospital with persistent asymptomatic microscopic haematuria between January 2016 to December 2023 was conducted. The demographic characteristics, kidney pathology and gene results of 135 individuals were analyzed. One hundred and thirty-five individuals were divided into 2 groups (positive group and negative group) according to family history of glomerulogenic hematuria in first-degree relatives. The differences of hematuria remission, proteinuria and gene variation were compared between the 2 groups. Two independent sample t test, Wilcoxon rank sum test, Pearson Chi-square, Yates′ corrected Chi-squared test or Fisher exact test were used for comparison between groups. Results:All 135 children, with 48 males and 87 females, were 8.5 (6.5, 9.5) years old at first presentation. Kidney biopsy was performed in 73 cases (54.1%). Kidney pathology showed mild lesions in 41 cases (56.2%), thin basement membrane disease (TBMD) in 24 cases (32.9%), typical pathological features of Alport syndrome in 5 cases (6.8%), and other manifestations in 3 cases (4.1%). The positive group comprised 52 individuals, whereas the negative group consisted of 83 individuals. The positive group demonstrated a higher susceptibility in proteinuria and gene variation, while the negative group exhibited a greater rate of hematuria remission ( χ2=5.00, 5.27, 8.52, all P<0.05). Whole exome sequencing was performed in 80 individuals and 18 individuals (22.5%) had a pathogenic or likely pathogenic variant in COL4A3-5. COL4A5 was the most common gene afected, accounting for 11 cases. The 135 individuals were followed up for 4.2 (2.9, 5.1) years, of which 31 cases (22.9%) had complete hematuria remission at 2.1 (1.4, 2.7) years. Up to March 2024, there were also 7 individuals (5.2%) with varying degrees of proteinuria, and 3 individuals (2.2%) with proteinuria progressed to chronic kidney insufficiency. Conclusions:The most common kidney pathological types in children with persistent asymptomatic microscopic hematuria are minor lesions and TBMD. Children with microscopic hematuria whose first-degree relatives have a family history of hematuria are more likely to have proteinuria and gene variants. COL4A3-5 genetic screening could be considered a priority in these children.

Objective:To explore clinical and genetic features of persistent asymptomatic microscopic hematuria in children.Methods:A retrospective case analysis of 135 individuals admitted to Xi ′an Children′s Hospital with persistent asymptomatic microscopic haematuria between January 2016 to December 2023 was conducted. The demographic characteristics, kidney pathology and gene results of 135 individuals were analyzed. One hundred and thirty-five individuals were divided into 2 groups (positive group and negative group) according to family history of glomerulogenic hematuria in first-degree relatives. The differences of hematuria remission, proteinuria and gene variation were compared between the 2 groups. Two independent sample t test, Wilcoxon rank sum test, Pearson Chi-square, Yates′ corrected Chi-squared test or Fisher exact test were used for comparison between groups. Results:All 135 children, with 48 males and 87 females, were 8.5 (6.5, 9.5) years old at first presentation. Kidney biopsy was performed in 73 cases (54.1%). Kidney pathology showed mild lesions in 41 cases (56.2%), thin basement membrane disease (TBMD) in 24 cases (32.9%), typical pathological features of Alport syndrome in 5 cases (6.8%), and other manifestations in 3 cases (4.1%). The positive group comprised 52 individuals, whereas the negative group consisted of 83 individuals. The positive group demonstrated a higher susceptibility in proteinuria and gene variation, while the negative group exhibited a greater rate of hematuria remission ( χ2=5.00, 5.27, 8.52, all P<0.05). Whole exome sequencing was performed in 80 individuals and 18 individuals (22.5%) had a pathogenic or likely pathogenic variant in COL4A3-5. COL4A5 was the most common gene afected, accounting for 11 cases. The 135 individuals were followed up for 4.2 (2.9, 5.1) years, of which 31 cases (22.9%) had complete hematuria remission at 2.1 (1.4, 2.7) years. Up to March 2024, there were also 7 individuals (5.2%) with varying degrees of proteinuria, and 3 individuals (2.2%) with proteinuria progressed to chronic kidney insufficiency. Conclusions:The most common kidney pathological types in children with persistent asymptomatic microscopic hematuria are minor lesions and TBMD. Children with microscopic hematuria whose first-degree relatives have a family history of hematuria are more likely to have proteinuria and gene variants. COL4A3-5 genetic screening could be considered a priority in these children.

Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*

Objective To investigate the genetic causes of auditory neuropathy with optic atrophy in a family.Methods The proband's medical history and family history were inquired in detail,and relevant clinical examina-tions were performed to confirm the diagnosis of auditory neuropathy with optic atrophy,and the genetic pedigree of the family was drawn.Peripheral blood of proband(Ⅲ-7)was collected for whole exome sequencing,and the patho-genicity of the detected mutations were interpreted.Blood samples of proband's wife(Ⅲ-8),eldest daughter(Ⅳ-7),second daughter(Ⅳ-9)and son(Ⅳ-10)were tested for mutation sites by Sanger sequencing.Combined with clinical manifestations and examination results,the family was studied.Results The genetic pattern of this family was autosomal dominant.The proband showed decreased visual acuity at the age of 19,bilateral sensorineural deaf-ness at the age of 30,and decreased speech recognition rate.Among 20 members of the family of 5 generations,10(2 deceased)showed similar symptoms of hearing and visual impairment.Proband(Ⅲ-7),eldest daughter(Ⅳ-7)and son(Ⅳ-10)underwent relevant examination.Pure tone audiometry showed bilateral sensorineural deafness.ABR showed no response bilaterally.The 40 Hz AERP showed no response in both ears.OAE showed responses in some or all of the frequencies.No stapedial reflex was detected.The eye movement of Ⅲ-7 and Ⅳ-10 were reasona-ble in all directions,and color vision was normal.Ocular papilla atrophy was observed in different degrees in fundus examination.OCT showed thinning of optic disc nerve fibers in both eyes,and visual evoked potential showed pro-longed P100 wave peak.They were diagnosed as hereditary auditory neuropathy with optic atrophy.A mutation of the OPA1 gene c.1334G＞A(p.Arg445His,NM_015560.2)at a pathogenic locus on chromosome 3 was detected by whole exon detection in Ⅲ-7.The results of generation sequencing analysis showed that the OPA1 gene c.1334G＞A(p.Arg445His,NM_015560.2)mutation of chromosome 3 was also found in Ⅳ-7 and Ⅳ-10.Meanwhile,the gen-otypes of Ⅲ-8 and Ⅳ-9 were wild homozygous,that is,no mutation occurred.Conclusion The OPA1 c.1334G＞A(p.Arg445His,NM_015560.2)mutation site might be the pathogenic mutation in this family.

Purpose To explore the cardioprotective effect of cang-ai volatile oil(CAVO)on rats with cardiac function impairment model under low-pressure and low-oxygen environment in Tibet Plateau based on 7.0T cardiovascular magnetic resonance(CMR)imaging.Materials and Methods Forty SD rats were randomly divided into the normal group,the high altitude model group,the CAVO-treated group and the rhodiola rosea-treated group,with 10 rats in each group.Except for the normal group,the rats in other groups were transferred from the plain(500 m above sea level)to the Tibet Plateau(4 250 m above sea level)for two months,and then administered with the corresponding drugs by gavage for 14 d.The left ventricle function was measured by using a 7.0T high-field strength CMR and myocardial strain was analysed by using tissue tracing technique.HE staining was used to observe the morphology of cardiomyocytes,Masson staining to observe interstitial fibrosis,wheat germ agglutinin staining to observe cardiomyocyte hypertrophy,and transmission electron microscopy to observe the morphological changes of mitochondria in each group.Serum levels of creatine kinase,creatine kinase isoenzyme,lactate dehydrogenase,cardiac troponin T,superoxide dismutase,malondialdehyde and glutathione peroxidase were detected.Intracellular reactive oxygen species levels were detected using flow cytometry.Results The left ventricular ejection fraction of rats in the CAVO-treated group was higher than that of the high altitude model group[(66.61±1.38)％vs.(60.94±3.21)％;t=3.969,P=0.032];meanwhile,the global circumferential strain of the left ventricle in the CAVO-treated group was higher than that of the high altitude model group(-25.68±1.30 vs.-22.84±1.17;t=3.967,P=0.003).HE,Masson and wheat germ agglutinin staining showed hypertrophy and necrosis as well as interstitial fibrosis and ultrastructural disruption of cardiomyocytes in the high altitude model group,which improved after CAVO treatment.The level of cardiac troponin T in the serum of rats with CAVO treatment group was significantly decreased compared with that of the high altitude model group[(314.03±20.05)pg/ml vs.(518.30±18.13)pg/ml;1=13.090,P=0.001].Conclusion CAVO treatment can reduce cardiac injury caused by low-pressure hypoxia in high altitude,and its effect can be detected dynamically and non-invasively by 7.0T high-field strength CMR.

Objective To establish a hypobaric hypoxia rat model in a real high-altitude environment,to investigate the effects of the real high-altitude environment on rat bone mass and bone microstructure using multiple methods such as Micro CT,blood biochemistry,and pathology,and to explore the potential mechanisms involved.Methods Sprague Dawley(SD)rats were transported to the Yushu Plateau Laboratory(at 4250 m above sea level)in Qinghai Province and kept there for 4,or 8,or 18 months.These groups were designated as H-4,H-8,and H-18,respectively.Upon completion of the high-altitude exposure,these animals were transported to the Molecular Imaging Laboratory,West China Hospital,Sichuan University(at 500 m above sea level)in Chengdu for relevant testing and comparison with the control animals raised in a low-altitude environment for the same durations(designated L-4,L-8,and L-18).The tests performed included blood biochemistry,Micro CT imaging,and pathological assessments such as ELISA,Western blot,and HE and TRAP staining.Results Compared with that of the control group,the body mass of rats in the H-4 and H-18 groups decreased significantly(H-4 group vs.L-4 group:[513.75±35.10]g vs.[649.18±60.03]g,P＜0.01;H-18 group vs.L-18 group:[535.58±66.65]g vs.[670.86±44.96]g,P＜0.01).The serum Ca2+concentration was higher in the H-8 group and H-18 group compared to that in the control group(H-8 group vs.L-8 group:[2.48±0.09]mmol/L vs.[2.38±0.07]mmol/L,P＜0.05;H-18 group vs.L-18 group:[2.55±0.11]mmol/L vs.[2.13±0.27]mmol/L,P＜0.05).No statistically significant difference was observed in the concentration of P3+.Bone metabolism indicator cross-linked carboxy-terminal telopeptide of type Ⅰ collagen(CTX-Ⅰ)was significantly increased in all high-altitude groups compared to the low-altitude groups(H-4 group vs.L-4 group:[1.44±0.08]ng/mL vs.[0.70±0.13]ng/mL,P＜0.01;H-8 group vs.L-8 group:[1.52±0.10]ng/mL vs.[0.75±0.10]ng/mL,P＜0.01;H-18 group vs.L-18 group:[2.70±0.13]ng/mL vs.[1.94±0.15]ng/mL,P＜0.01).In addition,CT results showed a decrease in bone volume fraction of trabecular bone in the three high-altitude groups(H-4 group vs.L-4 group:[7.48±2.35]％vs.[10.40±2.93]％,P＜0.05;H-8 group vs.L-8 group:[7.17±2.68]％vs.[10.09±2.95]％,P＜0.05;H-18 group vs.L-18 group:[2.90±2.91]％vs.[8.68±4.11]％,P＜0.01),and increased trabecular separation in the three high-altitude groups(H-4 group vs.L-4 group:[0.70±0.12]mm vs.[0.60±0.06]mm,P＜0.05;H-8 group vs.L-8 group:[0.68±0.07]mm vs.[0.59±0.05]mm,P＜0.01;H-18 group vs.L-18 group:[0.80±0.09]mm vs.[0.70±0.09]mm,P＜0.05).TRAP staining showed an increase in osteoclasts in the H-4 and H-18 groups.Western blot results indicated an increase in the expression of receptor activator of nuclear factor-κB ligand(RANKL)and hypoxia inducible factor-1α(HIF-1α)in high-altitude environment,while the expression of osteoprotegerin(OPG)was inhibited.Conclusion The impact of high-altitude environment on rat femurs is characterized primarily by a reduction in trabecular bone mass and damage to bone microstructure.