Objective:To explore the genetic and clinical characteristics of Guizhou patients with enlarged vestibular aqueduct（EVA） syndrome through combined SLC26A4 variant analysis and clinical phenotype analysis. Methods:Seventy-two EVA patients underwent comprehensive genetic testing using a multiplex PCR-based deafness gene panel and next-generation sequencing（NGS）. The audiological and temporal bone imaging characteristics were compared across mutation subtypes. Results:A total of 27 pathogenic loci of SLC26A4 were detected in 72 patients, including c.919-2A>G in 79.2%（57/72）. A novel deletion（c.1703_1707+6del） was discovered. Among 65 cases, truncated mutations were 89.2%（58/65）, 52.3%（34/65）, 28（43.1%） and 7（10.8%）. No significant differences were observed in the midpoint diameter of the vestibular aqueduct and the incidence of incomplete partitioning typeⅡ（IP-Ⅱ） of the cochlea among the three groups of patients. Moreover, there was no difference in the midpoint diameter of different vestibular pipes or the combination with IP-Ⅱ. Conclusion:The most common mutation site of SLC26A4 in EVA patients in Guizhou is c.919-2A>G, though genotype-phenotype correlations remain elusive. The detection of 27 mutation sites and the discovery of new mutation sites suggested the precise diagnostic significance of NGS technology in EVA patients in Guizhou.
Humans ; Sulfate Transporters ; Vestibular Aqueduct/abnormalities* ; Mutation ; Membrane Transport Proteins/genetics* ; Hearing Loss, Sensorineural/genetics* ; Male ; Female ; Child ; Adolescent ; Child, Preschool ; Adult ; Young Adult ; Phenotype ; High-Throughput Nucleotide Sequencing

Objective To construct a risk prediction model for atherosclerosis(AS)in patients with rheumatoid arthritis(RA)based on Lasso-Logistic regression analysis and provide a scientific basis for individualized clinical intervention.Methods The retrospective clinical data were collected from 344 RA patients,including 86 patients with AS(RA+AS group)and 258 patients with without AS(RA group).The clinical characteristics and initial laboratory test results were compared between the two groups.Lasso regression was used to screen the key predictive variables,and Logistic regression was combined to construct the prediction mode.The discrimination of the model was evaluated through the receiver operating characteristic(ROC)curve and the area under the curve(AUC).The Hosmer-Lemeshow test was used to assess the calibration,and decision curve analysis was used to verify the clinical applicability of the model.Results Seven predictive variables were identified including RA disease duration,DAS28 score,C-reactive protein(CRP),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),fasting blood glucose(FBG)and hypertension.The risk prediction model for AS in RA patients was:Logit(P)=-2.674+0.605×RA disease duration+0.393×DAS28 score+0.310×CRP+1.346×TG-2.289×HDL-C+0.679×FBG+0.711×hypertension.The AUC of the model was 0.965(95%CI:0.943-0.987),and the Hosmer-Lemeshow test showed χ2=0.547,P=1.000,indicating good discrimination and calibration.Clinical decision curve analysis showed that the probability threshold ranged from 7%to 92%,demonstrating high clinical applicability.Conclusion The AS risk prediction model constructed in this study for RA patients can effectively identify high-risk individuals,supporting the development of personalized prevention and treatment strategies.

Objective To construct a risk prediction model for atherosclerosis(AS)in patients with rheumatoid arthritis(RA)based on Lasso-Logistic regression analysis and provide a scientific basis for individualized clinical intervention.Methods The retrospective clinical data were collected from 344 RA patients,including 86 patients with AS(RA+AS group)and 258 patients with without AS(RA group).The clinical characteristics and initial laboratory test results were compared between the two groups.Lasso regression was used to screen the key predictive variables,and Logistic regression was combined to construct the prediction mode.The discrimination of the model was evaluated through the receiver operating characteristic(ROC)curve and the area under the curve(AUC).The Hosmer-Lemeshow test was used to assess the calibration,and decision curve analysis was used to verify the clinical applicability of the model.Results Seven predictive variables were identified including RA disease duration,DAS28 score,C-reactive protein(CRP),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),fasting blood glucose(FBG)and hypertension.The risk prediction model for AS in RA patients was:Logit(P)=-2.674+0.605×RA disease duration+0.393×DAS28 score+0.310×CRP+1.346×TG-2.289×HDL-C+0.679×FBG+0.711×hypertension.The AUC of the model was 0.965(95%CI:0.943-0.987),and the Hosmer-Lemeshow test showed χ2=0.547,P=1.000,indicating good discrimination and calibration.Clinical decision curve analysis showed that the probability threshold ranged from 7%to 92%,demonstrating high clinical applicability.Conclusion The AS risk prediction model constructed in this study for RA patients can effectively identify high-risk individuals,supporting the development of personalized prevention and treatment strategies.

Objective This study aims to discuss the research hotspot and development trend in the field of polycystic ovary syndrome(PCOS)through bibliometric statistics and visual analysis of long noncoding RNA(lncRNA)related studies.Methods Utilizing the Web of Science core database as the literature data source,we searched for PCOS lncRNA-related literature from 2015 to 2023.CiteSpace software was used to conduct a visual analysis,including the annual distribution,citation trends,countries,institutions,funding sources and key words,as well as co-occurrence and cluster analysis of key words.Results The visual analysis of 108 PCOS lncRNA literature revealed that China was the country with the highest number of publications.The first contributing institution was the Shandong University.The national natural science fund of China gave the biggest funding.The keyword cluster analysis suggested that PCOS lncRNA signal pathway regulation,related receptor activators,and the expression of regulatory factors were the research hotspots in ovary syndrome lncRNA research.Conclusion LncRNA related regulatory factors,bioinformatics analysis,and gene transcription in PCOS are new targetsfor PCOS treatment,providing valuable insights for clinical therapy and new strategies for the development of PCOS-related pharmaceuticals.

Objective:To investigate the incidence and influencing factors of olfactory dysfunction in postoperative laryngeal cancer patients.Methods:From November 2022 to August 2023, totally 76 postoperative laryngeal cancer patients admitted to the Department of Head and Neck Surgery at Zhejiang Cancer Hospital were selected by convenience sampling. General information questionnaire and the T&T olfactometer were used for assessment. Multiple linear regression analysis was conducted to explore the influencing factors of olfactory dysfunction in postoperative laryngeal cancer patients.Results:The T&T olfactometer score for the 76 postoperative laryngeal cancer patients at two weeks post-surgery was (4.74±0.93). Olfactory dysfunction occurred in 76 patients, with 25 cases of anosmia, 33 cases of severe hyposmia, 17 cases of moderate hyposmia, and one case of mild hyposmia. Multiple linear regression analysis revealed that age, educational level, surgical method, and smoking history were influencing factors for olfactory dysfunction in these postoperative laryngeal cancer patients.Conclusions:Postoperative laryngeal cancer patients experience varying degrees of olfactory dysfunction. Healthcare providers should pay special attention to older patients, those with lower educational levels, those undergoing total laryngectomy, and those with a history of smoking. Designing practical olfactory rehabilitation programs and implementing early olfactory training methods can help reduce the incidence of olfactory dysfunction.

Objective:To explore the clinical characteristics and variant of CREBBP gene in a fetus with Rubinstein-Taybi syndrome (RSTS). Methods:A fetus with RSTS diagnosed at the Third Affiliated Hospital of Zhengzhou University in August 2022 was selected as the study subject. Clinical data, amniotic fluid sample of the fetus and peripheral blood samples of its parents were collected for whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:Foot malformation, cerebellar vermis agenesis, brain agenesis, polysyndactyly of the big toes and other phenotypes were found by prenatal ultrasound. WES revealed that the fetus has harbored a heterozygous c. 4684G>T (p.E1562*) variant in exon 28 of the CREBBP gene (NM_004380.3), which was de novo in origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic (PVS1+ PS2_Moderate+ PM2_Supporting). After genetic counseling, the couple had opted to terminate the pregnancy and refused autopsy for the fetus. Conclusion:The c. 4684G>T (p.E1562*) variant of the CREBBP gene probably underlay the RSTS in this fetus. The newly discovered variant has enriched the mutational spectrum of the CREBBP gene and illustrated that WES is an efficient tool for the prenatal diagnosis of RSTS.

Objective To investigate the role of PRSS3/FGFR2 in immune escape of non-small cell lung cancer(NCLC)cells by using an A549 cell model which stably knocks down PRSS3 and overexpresses FGFR2 on the basis of PRSS3.Methods A549 cells were divided into sh-NC group,sh-PRSS3 group,sh-PRSS3+ov-NC group and sh-PRSS3+ov-FGFR2 group,and were transfected with corresponding lentiviruses respectively.Western blot was used to detect the expression of PRSS3,FGFR2,p-JAK2,JAK2,p-STAT3,STAT3 and PD-L1 in A549 cells of different groups;CCK-8 was used to detect the proliferation of theses A549 cells;Transwell was used to detect the migration and invasion ability of theses A549 cells.Furhtermore,CD8+T cells were co-cultured with these A549 cells,and trypan blue staining and flow cytometry were used to detect the viability and apoptosis rate of the CD8+T cells,respectively.The levels of sPD-L1,IFN-γ,TNF-α,granzyme B and perforin in the supernatant of co-cultured cells were detected by ELISA.Results Compared with sh-NC group,sh-PRSS3 group and sh-PRSS3+ov-NC group demonstrated lower expressions of PRSS3,FGFR2,p-JAK2,p-STAT3 and PD-L1 proteins,as well as lower levels of proliferation activity,the number of migrating cells and the number of invading cells of A549 cells.Furthermore,in the supernatant of co-culture medium of sh-PRSS3 and sh-PRSS3+ov-NC groups,the viability of CD8+T cells was increased,the levels of IFN-γ,TNF-α,granzyme B and perforin were elevated,while the apoptosis of CD8+T cells and the level of sPD-L1 were decreased,as compared with sh-NC group.In the sh-PRSS3+ov-FGFR2 group,all these differences mentoned above were mitigated,which means the effects of sh-PRSS3 were antagonized by overexpression of FGFR2.Conclusion PRSS3 induces immune escape of NSCLC cells by up-regulating FGFR2 expression,and its mechanism may be related to the up-regulation of PD-L1 expression induced by activating JAK2/STAT3 signaling pathway.

This study was designed to explore whether overexpression of HMBOX1 inhibits COPD-induced infiltration and pulmonary macrophage activation by regulating NF-κB/CCL2 signaling pathway.Forty Wistar rats were randomly divided into control group,chronic obstructive pulmonary disease group(COPD group),COPD+control overexpression group(COPD+ov-NC group)and chronic COPD+HMBOX1 overexpression group(COPD+ov-HMBOX1 group),with 10 rats in each group.The COPD model was established by continuous cigarette incense and intermittent intratracheal injection of lipopolysaccharide,wihle the HMBOX1 overexpression treatmet was carried out by intratracheal instillation of HMBOX1 overexpressing adenovirus.Western blot was used to detect the expression of HMBOX1,p-NF-κB and NF-κB proteins in lung tissue of rats;RT-qPCR was used to detect the mRNA expression of HMBOX1 and CCL2 in rat lung tissue;HE staining was used to observe the pathological changes of lung tissue in rats;ELISA was applied to detect the levels of TNF-α,MIP-2,IL-1β and IL-10 in serum and BALF of rats.Furthermore,the ratio of CD11b+F4/80+cells in lung tissue of rats was detected by immunofluorescence,while the ratio of F4/80+MHC Ⅱ+cells and F4/80+CD80+cells in lung tissue of rats was detected by flow cytometry.In control group,the alveolar structure of rats was intact,and no inflammatory cell infiltration was found.In COPD group and COPD+ov-NC group,a large number of inflammatory cells infiltrated into the lung tissue and alveolar structure was damaged.In COPD+ov-HMBOX1 group,there were fewer inflammatory cells infiltrated in lung tissue,and the damage of alveolar structure was alleviated.Compared with the control group,the mRNA and protein expression of HMBOX1 in lung tissue,the levels of IL-10 in serum and BALF,the levels of TNF-α,MIP-2 and IL-1β in serum and BALF,CD11b+F4/80+cells,F4/80+MHC Ⅱ+cells and F4/80+CD80+cells in lung tissue of rats in COPD and COPD+ov-NC groups were significantly decreased.HMBOX1 overexpression could revers the changes mentioned above in the two COPD groups.Taken together,overexpression of HMBOX1 can alleviate COPD-induced airway inflammation and lung injury,and its mechanism may be related to inhibiting infiltration and abnormal activation of macrophages in lung tissue mediated by activation of NF-κB/CCL2 signaling pathway.

Objective To establish a rat model of chronic obstructive pulmonary disease(COPD),to explore whether berberine can improve the imbalance of Th17/Treg cells induced by COPD by regulating NF-κB/LCN2 signaling pathway.Methods Fifty Wistar rats were randomly divided into Control group(control group),chronic obstructive pulmonary disease group(COPD group),Berberine group(Berberine group),berberine+overexpression control group(Berberine+ov-NC group)and berberine+overexpression LCN2 group(Berberine+ov-LCN2 group),with 10 rats in each group.Western blotting was used to detect the expression of p-NF-κB,LCN2,IL-17 and IL-10 in lung tissue.HE staining was used to observe the pathological changes of lung tissue in rats.The ratio of Th17/Treg cells in peripheral blood of rats was detected by flow cytometry.The levels of IL-17 and IL-10 in serum of rats were detected by ELISA.Results In the control group,the lung tissue structure was intact;compared with the control group,the lung tissue structure of rats in COPD group was damaged,the expressions of p-NF-κB and LCN2 protein in lung tissue were significantly increased,the proportion of Th17 cells in peripheral blood was significantly increased,the proportion of Treg cells was significantly decreased,the level of IL-17 in serum and the expression of IL-17 protein in lung tissue were significantly increased,and the level of IL-10 in serum and the expression of IL-10 protein in lung tissue were significantly decreased(P<0.05).Compared with COPD group,Berberine group and Berberine+ov-NC group improved lung tissue injury,significantly decreased the expression of p-NF-κB and LCN2 proteins in lung tissue,significantly decreased the proportion of Th17 cells in peripheral blood,significantly increased the proportion of Treg cells,significantly decreased the level of IL-17 in serum and the expression of IL-17 protein in lung tissue(P<0.05).Compared with Berberine group and Berberine+ov-NC group,Berberine+ov-LCN2 group has more lung injury,higher expression of LCN2 protein in lung tissue,higher proportion of Th17 cells in peripheral blood,lower proportion of Treg cells,higher levels of IL-17 in serum and IL-17 protein expression in lung tissue,and higher levels of IL-10 in serum and IL-10 protein in lung tissue(P<0.05).Conclusion Berberine treatment can improve lung injury and Th17/Treg cell imbalance in COPD rats by down-regulating the expression of LCN2,and its mechanism may be related to the inhibition of NF-κB/LCN2 signaling pathway.

Objective To investigate the role of PRSS3/FGFR2 in immune escape of non-small cell lung cancer(NCLC)cells by using an A549 cell model which stably knocks down PRSS3 and overexpresses FGFR2 on the basis of PRSS3.Methods A549 cells were divided into sh-NC group,sh-PRSS3 group,sh-PRSS3+ov-NC group and sh-PRSS3+ov-FGFR2 group,and were transfected with corresponding lentiviruses respectively.Western blot was used to detect the expression of PRSS3,FGFR2,p-JAK2,JAK2,p-STAT3,STAT3 and PD-L1 in A549 cells of different groups;CCK-8 was used to detect the proliferation of theses A549 cells;Transwell was used to detect the migration and invasion ability of theses A549 cells.Furhtermore,CD8+T cells were co-cultured with these A549 cells,and trypan blue staining and flow cytometry were used to detect the viability and apoptosis rate of the CD8+T cells,respectively.The levels of sPD-L1,IFN-γ,TNF-α,granzyme B and perforin in the supernatant of co-cultured cells were detected by ELISA.Results Compared with sh-NC group,sh-PRSS3 group and sh-PRSS3+ov-NC group demonstrated lower expressions of PRSS3,FGFR2,p-JAK2,p-STAT3 and PD-L1 proteins,as well as lower levels of proliferation activity,the number of migrating cells and the number of invading cells of A549 cells.Furthermore,in the supernatant of co-culture medium of sh-PRSS3 and sh-PRSS3+ov-NC groups,the viability of CD8+T cells was increased,the levels of IFN-γ,TNF-α,granzyme B and perforin were elevated,while the apoptosis of CD8+T cells and the level of sPD-L1 were decreased,as compared with sh-NC group.In the sh-PRSS3+ov-FGFR2 group,all these differences mentoned above were mitigated,which means the effects of sh-PRSS3 were antagonized by overexpression of FGFR2.Conclusion PRSS3 induces immune escape of NSCLC cells by up-regulating FGFR2 expression,and its mechanism may be related to the up-regulation of PD-L1 expression induced by activating JAK2/STAT3 signaling pathway.