ObjectiveTo investigate the mRNA expression profile and clinical significance of the RAB24 gene in hepatocellular carcinoma （HCC） based on multi-source database including The Cancer Genome Atlas （TCGA） and clinical cohort validation. MethodsData from TCGA （n=424） were utilized to evaluate the prognostic value of RAB24 expression via Kaplan-Meier analysis and Cox regression models. An independent clinical cohort comprising 90 HCC and 30 paracancerous tissues was established to validate protein expression and histopathological features through immunohistochemistry （IHC） and routine histological staining. The TIMER database was employed to analyze the association between RAB24 and the tumor microenvironment （TME）. Furthermore， integrated with LinkedOmics， GSEA and GSVA algorithms were used to quantitatively characterize the core molecular pathways regulated by RAB24. ResultsThe mRNA expression level of RAB24 in HCC was significantly higher than that in non-tumor liver tissues （P<0.05）. Kaplan-Meier analysis revealed that the overall survival （OS） of patients in the RAB24 low-expression group was significantly longer compared to the high-expression group （HR=1.570， Log-rank P=0.01）. Cox proportional hazards regression analysis demonstrated that RAB24 expression （HR=1.480， P=0.028）， T stage （HR=2.949， P<0.001）， M stage （HR=4.077， P=0.017）， and tumor residual status （HR=2.317， P<0.001） were independent prognostic factors for HCC. RAB24 expression was highly correlated with alpha-fetoprotein （AFP） levels and immune infiltration levels （P<0.01）， and showed a significant， low-level positive correlation with the TP53 gene （Rho=0.31， P<0.01）. Quantitative analysis of the RAB24 protein in clinical samples validated that RAB24 expression in HCC tissues was significantly elevated compared to non-tumor liver tissues （P<0.05）. TIMER analysis confirmed that RAB24 expression was highly associated with the infiltration of multiple immune cells （such as B cells and NK cells）， as well as stromal and tumor purity scores in tumor tissues （P<0.01）. GSEA/GSVA enrichment revealed that the P53， MYC， and mTORC1 signaling pathways were activated in the RAB24 high-expression group； correlation analysis confirmed that the RAB24 expression level exhibited a significant， low-level positive correlation with the ribosome pathway activity score （r=0.23， P<0.01）. ConclusionsThe RAB24 gene， as a poor prognostic factor for HCC patients， is a potential biomarker for HCC diagnosis， treatment and prognosis. The study is expected to lay a theoretical foundation for the precise diagnosis and treatment of clinical HCC.

Objective To investigate the effect of miR-132-3p on epithelial-mesenchymal transition(EMT)of trophoblast cells through targeted regulation of the SIRT1/NF-κB pathway.Methods Placental tissues from 37 pre-eclampsia(PE)patients who underwent cesarean section in Zhengzhou Women&Infants Hospital and 37 normal pregnant women who underwent cesarean section were collected as the research group and control group,respectively.RT-qPCR was performed to detect the expression of miR-132-3p and SIRT1 in tissues.During the logarithmic pro-liferation phase,HTR-8/SVneo cells were grouped into inhibitor negative group,miR-132-3p inhibitor group,mimic negative group,miR-132-3p overexpression group,miR-132-3p inhibitor+interference negative group,and miR-132-3p inhibitor+interference SIRT1 group,with untransfected HTR-8/SVneo cells as the blank group.CCK-8,Transwell assay,scratch assay,and RT-qPCR were used to detect the proliferation,invasion,migration,and changes in miR-132-3p and SIRT1 mRNA expression of HTR-8/SVneo cells.Western blot was used to detect the expression levels of EMT related proteins and SIRT1,NF-κB proteins.Dual luciferase was used to validate the targeting relationship between miR-132-3p and SIRT1.Results The level of miR-132-3p in the research group was significantly higher than that in the control group,while the SIRT1 mRNA was lower(P＜0.05).The level of miR-132-3p,E-cadherin,p-NF-κBp65/NF-κBp65 in the miR-132-3p inhibitor group was lower than those in the inhib-itor negative group and blank group.The proliferation rate,invasion cell number,migration rate,level of vimentin,N-cadherin,and SIRT1 were all higher(P＜0.05).The miR-132-3p,E-cadherin,p-NF-κBp65/NF-κBp65 in the miR-132-3p overexpression group were higher than those in the mimic negative group,the proliferation rate,inva-sion number,migration rate,vimentin,N-cadherin,and SIRT1 were lower(P＜0.05).The level of E-cadherin and p-NF-κBp65/NF-κBp65 in the miR-132-3p inhibitor+interference SIRT1 group was higher than those in the miR-132-3p inhibitor+interference negative group(P＜0.05),the proliferation rate,invasion number,migration rate,vimentin,N-cadherin,and SIRT1 expression were lower(P＜0.05).MiR-132-3p targeted regulation of SIRT1 expression(P＜0.05).Conclusions Inhibition of miR-132-3p promotes the EMT,proliferation,invasion,and migration of trophoblast cells by targeting and activating the SIRT1/NF-κB pathway.

Objective To analyze the PLCβ4 gene mRNA expression and its clinical significance in hepatocellular carcinoma (HCC) based on TCGA database. Methods Based on the data on 424 clinical samples (including 374 cases of HCC tissues and 50 cases of nontumor liver tissues) in the TCGA database, Kaplan–Meier method, Cox regression analysis, and immune infiltration analysis were performed to evaluate the relationship between PLCβ4 gene and the clinical characteristics and survival prognosis of HCC patients. Correlation analysis between PLCβ4 gene and 24 types of immune cells was applied to investigate the relationship between PLCβ4 gene and immune cell infiltration and mRNA expression level of TP53 gene, a high-frequency mutation gene in HCC. In addition, paraffin sections of highly, moderately, and poorly differentiated tumor tissues and normal liver tissues from HCC patients were collected. The histopathological observation was carried out via HE staining method, and the expression levels of PLCβ4 and Ki-67 proteins in each clinical sample were verified through the immunohistochemical method. Results The expression level of PLCβ4 gene in HCC was significantly higher than that in normal tissues (P<0.01), and all patients in the PLCβ4 high-expression group had a significantly longer overall survival than those in the low-expression group (P<0.05), which suggested that PLCβ4 substantially affected the prognosis of HCC patients. Correlation analysis showed that the expression level of PLCβ4 gene was highly correlated with immune cell infiltration and the expression level of TP53 gene. As verified by clinical sample experiments, HE staining experiments and immunohistochemical results revealed that PLCβ4 gene expression in HCC tissue samples was significantly higher than that in normal tissues (P<0.001), and it was negatively correlated with the degree of differentiation. Conclusion PLCβ4 may serve as an independent prognostic factor in HCC and is expected to be a novel molecular target for HCC treatment.

Thirteen novel diterpenoids, comprising seven tiglianes (walliglianes G-M, 1-7), four rhamnofolanes (wallinofolanes A-D, 8-11), and two daphnanes (wallaphnanes A and B, 12 and 13), together with two known rhamnofolane diterpenoids (euphorwallside H and euphorwallside I, 14 and 15), were isolated and characterized from Euphorbia wallichii(E. wallichii). The chemical structures of these compounds were elucidated through nuclear magnetic resonance (NMR), mass spectrometry (MS), and quantum chemical calculations. Compounds 9 and 11 demonstrated protective effects against H₂O₂-induced BV-2 microglial cell damage. Molecular docking analyses indicated that compound 9 exhibited binding affinity to the anti-oxidant-related targets HMGCR, GSTP1, and SHBG.
Euphorbia/chemistry* ; Antioxidants/isolation & purification* ; Diterpenes/isolation & purification* ; Molecular Structure ; Mice ; Molecular Docking Simulation ; Animals ; Hydrogen Peroxide/toxicity* ; Cell Line ; Microglia/drug effects*

Objective:To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband.Methods:A Chinese pedigree which opted elective abortion at the Women and Children′s Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). Results:Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c. 7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c. 1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. Conclusion:This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.

Objective:To determine the prevalence and associates of hyperuricemia (HUA) among the middle- and older-aged population of the island and mountainous areas in Taizhou City of Zhejiang Province.Methods:A cross-sectional study was conducted on individuals aged 45 and above in the island and mountainous area of Taizhou City. The study included questionnaires, physical examinations, and laboratory tests. Data were primarily collected on sociodemographic characteristics, chronic disease history, lifestyle factors, waist circumference, blood pressure, and serum uric acid levels. The association between hyperuricemia and these factors was analyzed by logistic regression.Results:A total of 971 individuals were included in the study, comprising 468 from island and 503 from mountainous area. The prevalence of hyperuricemia was 17.9%, with a significantly higher prevalence in the island area (25.6%) compared to the mountainous area (10.7%). Stratified by gender and age, differences in the prevalence of hyperuricemia between island and mountainous areas were observed in males aged 55 to 64, females aged 45 to 54, 55 to 64, 75 and above (all P<0.05). Multiple logistic regression analysis showed that high intake of sea food (>3 times/week) was positively associated with hyperuricemia ( OR=2.10, 95% CI:1.33-3.34). Furthermore, separate regionally stratified logistic regression analyses showed that in the island area, male gender ( OR=3.15, 95% CI:1.78-5.66), central obesity ( OR=2.38, 95% CI:1.46-3.93), and hypertriglyceridemia ( OR=2.00, 95% CI:1.22-3.30) were positively correlated with hyperuricemia (all P<0.05). In the mountainous area, the age group of 65 and above ( OR=3.50, 95% CI:1.09-12.50), male ( OR=6.79, 95% CI:2.87-17.81), those employed in enterprises and institutions ( OR=6.57, 95% CI:1.92-23.73) and hypertension ( OR=3.68, 95% CI:1.66-8.87) were positively correlated with hyperuricemia (all P<0.05). Conclusions:The prevalence of hyperuricemia among the middle- and older-aged population in the island of Taizhou City is significantly higher than that in the mountainous areas. Targeted comprehensive behavioral interventions such as routine screening of chronic diseases, low-fat diet, alcohol control, reduced seafood intake, enhanced exercise, weight management, and blood pressure control are warranted.

Objective To investigate the predictive value of the mini-fluid challenge test in elderly orthope-dic patients for post-spinal anesthesia hypotension.Methods Seventy-two elderly patients who underwent elective hip or knee replacement surgery were rigorously screened according to predefined inclusion and exclusion criteria.All patients were scheduled for subarachnoid block anesthesia.Subjects were grouped based on changes in blood pressure within 15 minutes of assuming a supine position following single-shot lumbar anesthesia.According to previ-ously established definitions of hypotension,they were categorized into either the hypotension group(H group)or the normal blood pressure group(N group).Prior to spinal anesthesia,a mini-fluid challenge test was conducted using noninvasive cardiac output monitoring to measure the change in stroke volume index(ΔSVI),and baseline circulatory data were recorded.Multivariate logistic regression analysis was employed to identify factors influencing outcomes in elderly patients undergoing orthopedic joint replacement surgery.Receiver operating characteristic(ROC)curves for ΔSVI were constructed,and the area under the curve(AUC)was calculated to evaluate its predic-tive performance.Results After spinal anesthesia,29 patients(40.27%)experienced hypotension.Compared with Group N,patients in Group H who experienced hypotension within 15 minutes while in a supine position were signifi-cantly older,had a higher proportion of ASA grade III,and a higher prevalence of hypertension(P<0.05).The analysis results indicated that ΔSVI was an independent influencing factor for post-lumbar anesthesia hypotension in elderly patients.ΔSVI demonstrated a sensitivity of 82.8%and a specificity of 81.4%in predicting post-spinal anes-thesia hypotension(PSAH)at a cut-off value of 0.805 or greater.There was a moderate positive linear correlation between the maximum decrease in systolic blood pressure(SBP)and ΔSVI(r=0.562,P<0.01).Conclusion The mini-fluid challenge test is an effective method for predicting hypotension in elderly orthopedic patients following spinal anesthesia.

OBJECTIVE: To explore the genetic etiology of a Chinese pedigree with recurrent Joubert syndrome with negative results by whole-exome sequencing in the prior proband. METHODS: Chinese pedigree which opted elective abortion at the Women and Children's Hospital Affiliated to Chongqing Medical University in December 2024 was selected as the study subject. Whole-genome sequencing was carried out on fetal tissue after termination of pregnancy. Candidate variants were validated by Sanger sequencing and interpreted, while non-coding variant was analyzed using in silico prediction tools. RNA sequencing and cDNA sequencing were conducted on fetal brain tissue. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.2024YL045-02). RESULTS: Both the fetus and the affected child were found to harbor compound heterozygous variants of the CEP290 gene, namely c.7341dup (p.Leu2448fs*8) (pathogenic, maternally inherited) and c.1523-408G>A (likely pathogenic, paternally inherited). Both in silico analysis and fetal brain RNA sequencing confirmed aberrant RNA splicing caused by the intronic variant. CONCLUSION This case has highlighted the value of combining whole-genome sequencing with RNA functional validation. Above results not only enriched the spectrum of CEP290 gene mutations but also underscored its diagnostic value in resolving complex prenatal cases, providing critical clues for the prenatal diagnosis and recurrence risk assessment in genetic counseling.
Female ; Humans ; Pregnancy ; Abnormalities, Multiple/genetics* ; Antigens, Neoplasm/genetics* ; Cell Cycle Proteins/genetics* ; Cerebellum/abnormalities* ; Cytoskeletal Proteins/genetics* ; Eye Abnormalities/genetics* ; Introns/genetics* ; Kidney Diseases, Cystic/diagnosis* ; Pedigree ; Retina/abnormalities* ; Sequence Analysis, RNA/methods* ; Whole Genome Sequencing/methods* ; Child

Objective To explore the value of cranial CT perfusion imaging(CTP)parameters combined with head and neck CT angiography(CTA)in assessing collateral circulation status and predicting prognosis in acute ischemic stroke(AIS).Methods A retrospective analysis was carried out on 83 AIS patients who were treated in Taikang Xianlin Drum Tower Hospital from June 2018 to June 2023.CTP and head and neck CTA examinations were performed within 24 hours after admission.Digital subtraction angiography is the gold standard for assessing collateral circulation status in AIS patients.The general information of these patients was collected,and the patient's prognosis was evaluated using the modified Rankin scale through telephone or outpatient follow-up 90 days after the occurrence of AIS.Pearson or Spearman correlation was used to analyze the correlation between collateral circulation assessment and CTP parameters and head and neck CTA scores.The value of CTP parameters and head and neck CTA scores in predicting the prognosis of AIS patients was discussed using multivariate Logistic regression.Moreover,the receiver operating characteristic(ROC)curve was used to analyze the predictive value of CTP parameters,head and neck CTA,and the combination for the prognosis of AIS patients.Results The cerebral blood volume(CBV),cerebral blood flow(CBF),and CTA score were higher,while mean transit time(MTT)and time to peak(TTP)were shorter in the good collateral circulation group than in poor collateral circulation group(P<0.05).The collateral circulation status in AIS patients was negatively correlated with CBV,CBF,and CTA score,while positively correlated with MTT and TTP(P<0.05).Compared with poor prognosis group,good prognosis group had higher CBV,CBF,CTA,and shorter MTT and TTP(P<0.05).Multivariate Logistic regression analysis identified MTT and TTP as risk factors for poor prognosis,and CBV,CBF,and CTA scores as protective factors for poor prognosis in AIS patients(P<0.05).The ROC results showed that the area under the curve(AUC)of CBV,MTT,CBF,TTP,CTA score and the combination to predict the prognosis of AIS patients were 0.897,0.864,0.835,0.920,0.918,and 0.979,respectively,showing better predictive performance of the combination than single index alone(Z=2.194,2.910,2.521,2.229,2.171;P<0.05).Conclusion CTP parameters combined with CTA may effectively assess collateral circulation in patients with AIS and is significant for prognosis prediction.

ObjectiveTo explore the construction and evaluation methods of a rat model of acute myocardial infarction（AMI） with Qi and Yin deficiency syndrome established by sleep deprivation combined with coronary artery ligation. MethodsThirty-six SD rats were randomly divided into a normal group（n=6）， a myocardial infarction group（model A group， n=10）， an acute sleep deprivation+myocardial infarction group（model B group， n=10）， and a chronic sleep deprivation+myocardial infarction group（model C group， n=10） according to body weight. Rats in the normal group were not treated， rats in the model A group underwent only ligation of the left anterior descending coronary artery， rats in the model B group were sleep deprived for 96 h and then underwent ligation of the left anterior descending coronary artery， and rats in the model C group were sleep deprived for an additional 48 h each week with a 24 h rest period as one cycle for three weeks on the basis of the model B group. After coronary artery ligation in the model C group， the first week was defined as the starting point of the first sleep deprivation cycle， and indexes were tested weekly for rats in each group for 3 weeks. Electrocardiogram was used to determine the ligation of the left anterior descending coronary artery in rats， and small animal echocardiography was used to evaluate the cardiac function. The levels of serum creatine kinase（CK）， creatine kinase isoenzyme（CK-MB）， lactate dehydrogenase（LDH）， cardiac troponin T（cTnT）， interleukin-18（IL-18）， and tumor necrosis factor-α（TNF-α） were detected by biochemical assays， and hematoxylin-eosin（HE） staining was used to evaluate the pathological changes of myocardial tissue in rats. The syndrome indicators of Qi and Yin deficiency were evaluated by general state and body weight， grip strength， facial temperature， paw temperature， rectal temperature， salivary flow rate， open field test， tongue color［red（R）， green（G）， and blue（B）］ values， pulse amplitude changes， and enzyme-linked immunosorbent assay（ELISA） for the detection of expression levels of cyclic adenosine monophosphate（cAMP）， cyclic guanosine monophosphate（cGMP）， rat serum corticotropin-releasing factor（CRF）， adrenocorticotropic hormone（ACTH）， triiodothyronine（T3）， tetraiodothyronine（T4）， and corticosterone（CORT） in serum. ResultsIn terms of disease indicators， compared with the normal group， the ST segment of the electrocardiogram in each model group was significantly elevated， the echocardiographic parameters were decreased， the contents of myocardial enzymes and inflammatory factors were increased（P<0.01）， and the myocardial tissue in the infarcted area was significantly damaged. In terms of syndrome indicators， compared with the normal group， the body weight of rats in the model B and C groups decreased at each time point， the grip strength of each model group decreased， the total distance traveled and the number of entries into the center in the open field test decreased， the immobility time increased， the facial and rectal temperatures of rats in the model B and C groups increased， the salivary flow rate of each model group decreased， the tongue color was bright red or light， the tongue body was dry or smooth like a mirror， lacking of moisture sensation， the R， G and B values of the tongue surface increased， the pulse amplitude changes decreased， and the contents of T3 and T4 increased， while the expressions of cAMP， CRF， ACTH and CORT in the model B and C groups increased（P<0.05， P<0.01）. ConclusionContinuous sleep deprivation for 96 h in a multi-platform method combined with coronary artery ligation can construct a rat model of AMI with Qi and Yin deficiency syndrome， and the syndrome manifestations can be maintained for 3 weeks.