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 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 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 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 integrate the best evidence of non-drug intervention of urinary incontinence in elderly women and to formulate practical recommendations.Methods:In this systematic review study, using “elderly woman”,“urinary incontinence”,“bladder training”,“pelvic floor muscle training”,“enuresis”,“leakage of urine” as the key words, the 6S evidence resource pyramid model was used to search in British Medical Journal best practice, Uptodate, World Health Organization, Guidelines International Network, National Institute for Health and Care Excellence, Chinese Medical Association, Scottish Intercollegiate Guideline Network, Registered Nurses Association of Ontario, Cochrane Library, The Joanna Briggs Institute (JBI), New Zealand Guidelines Group, Polish Society of Gynecologists and Obstetricians, PubMed, Embase, Medline, Web of Science, SinoMed, China National Knowledge Infrastructure, WanFang Data, etc. The evidence retrieved included evidence-based knowledge base resources, clinical practice guidelines, expert consensus, systematic review, etc. Data were retrieved from January 1, 2017 to May 1, 2022, and collated from May 2, 2022 to May 25, 2022. Two researchers independently evaluated the quality of literature and extracted data using the AGREE Ⅱ and JBI evidence-based health care center assessment tools. The JBI evidence-based health care center′s evidence pre-rating system and evidence recommendation rating system were applied to rank the evidence; and under the guidance of the evidence structure of JBI, the strength of evidence recommendation was determined and the best evidence was extracted and summarized in combination with the study group discussion and expert opinion.Results:A total of 9 articles were retrieved, including 7 guidelines and 2 systematic reviews; and 6 guidelines were classified as Grade A and 1 as grade B; both 2 systematic reviews were rated as Grade A; 84% (27/32) of the items were evaluated as “Yes”. Evidence were summarized as 34 pieces of best evidence from 6 dimensions, including “overall recommendation, evaluation of type and degree of urinary incontinence, lifestyle change, behavioral therapy, prevention of precipitating factors, intervention in special population”; the flow chart of screening, evaluation, special symptoms, life style and behavior therapy was combed, and the practical suggestions were formed.Conclusions:The overall quality of the literature on non-drug intervention of urinary incontinence in elderly women is high, and the level of evidence is high. Early identification of urinary incontinence types and assessment of disease severity, lifestyle changes, avoidance of predisposing factors and behavioral therapy are the key to non-drug treatment of urinary incontinence in those patients.

Objective:To understand the current status of the compliance of immunosuppressive drugs in patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to explore the influencing factors of their compliance.Methods:This study was a cross-sectional study. Using the convenient sampling method, a total of 238 allo-HSCT patients who were followed up in the Transplant Center of Taizhou Hospital of Zhejiang Province, Zhejiang Provincial Hospital of Traditional Chinese Medicine and the First Affiliated Hospital of Wenzhou Medical University from March 2020 to February 2021 were selected as the research objects. The self-designed general information questionnaire, Basel Assessment Scale for Immunosuppressive Drug Compliance, Chinese version of Medication Belief-Specific Scale, General Self-Efficacy Scale, Perceived Social Support Scale and Family Care Index Scale were used for on-site investigation of patients. Pearson correlation analysis was used to analyze the correlation between medication compliance and medication belief, self-efficacy, social support and family care of allo-HSCT patients. Multiple linear regression analysis was used to investigate the influencing factors of immunosuppressive medication compliance in allo-HSCT patients.Results:A total of 238 questionnaires were distributed in this study, and 228 valid questionnaires were finally recovered. The medication compliance score of allo-HSCT patients was (4.80±1.18) points. And 111 patients (48.68%, 111/228) had poor medication compliance, among which the incidence of not taking medication on time was the highest, accounting for 44.30% (101/238) . Multiple linear regression analysis showed that average monthly family income, rejection, medication belief, social support and family care were the influencing factors of medication compliance ( P< 0.05) . Conclusions:The immunosuppressant medication compliance of allo-HSCT patients is at a medium and low level. Average monthly family income, rejection reaction, medication belief, social support and family care are the main factors affecting the immunosuppressant medication compliance of allo-HSCT patients. Medical staff should formulate targeted intervention measures according to the influencing factors of medication compliance to improve the immunosuppressive medication compliance of allo-HSCT patients.

Objective:To construct a risk assessment scale for flap necrosis after flap transplantation in patients with head and neck tumors, so as to provide an effective reference for clinical implementation of flap necrosis risk screening.Methods:The overall study period was from December 2020 to June 2021. Based on evidence, the item pool of the risk assessment scale for flap necrosis after flap transplantation in patients with head and neck tumors was preliminarily established. Using the Delphi method, 16 experts engaged in flap transplantation were selected for three rounds of consultation, and the risk assessment scale for flap necrosis after flap transplantation in patients with head and neck tumors was initially established. The weights were given to all indicators through the paired comparison.Results:In the first round, 16 questionnaires were distributed, and 15 valid questionnaires were recovered. In the second and third rounds, 15 questionnaires were distributed, and 15 valid questionnaires were recovered. Expert authority coefficients of the three rounds of expert consultation were 0.851, 0.853 and 0.853, respectively. The Kendall coordination coefficients of the three rounds of expert consultation were 0.377, 0.302 and 0.302 ( P<0.05) . The final constructed risk assessment scale for flap necrosis after flap transplantation in patients with head and neck tumors included 3 first-level indicators, 8 second-level indicators, and 32 third-level indicators. The weights were assigned to each indicator by paired comparison, and the weights of the first-level indicator patient factor, treatment factor, and nursing factor were 0.36, 0.38, and 0.26, respectively. Conclusions:The risk assessment scale for flap necrosis after flap transplantation in patients with head and neck tumors constructed by combining evidence-based and Delphi method is highly scientific and reliable. Its clinical applicability and effectiveness can be further verified in the future clinical flap evaluation process.

Objective:To investigate the value of nomogram predictive model established by the risk factors of upper extremity venous thrombosis risk associated with peripherally venous inserted central catheter (PICC) in cancer patients.Methods:A total of 1 032 patients who underwent PICC insertion between January 2016 and March 2017 in Zhejiang Cancer Hospital were selected by using prospective cohort study and convenience sampling. Risk factors of upper extremity venous thrombosis risk associated with PICC in cancer patients were evaluated by using Cox regression model. The nomogram predictive model of upper extremity venous thrombosis risk associated with PICC insertion was constructed. Bootstrap method was used to complete the inside check, and figure calibration was used to verify the nomogram.Results:A multivariate Cox regression analysis showed that trombosis history ( HR = 27.82, 95% CI 8.17-94.88, P < 0.01) and hyperlipidemia ( HR = 3.01, 95% CI 1.31-6.93, P = 0.009) were independent risk factors for upper extremity venous thrombosis associated with PICC. The nomogram model C-index was 0.71 (95% CI 0.63-0.80) based on the above risk factors, which indicated that the nomogram had a good differentiation. The calibration curve for predicting the probability of upper extremity venous thrombosis risk associated with PICC within one week, two weeks and one month deviated slightly from the standard curve, suggesting that the model might overestimate the risk of upper extremity venous thrombosis associated with PICC in cancer patients. Conclusions:The nomogram model has a good predictive value and strong operability, which can be used to predict the probability of upper extremity venous thrombosis associated with PICC in cancer patients after PICC insertion. It can provide a reference for identifying the high-risk cancer patients and formulating proper therapeutic strategies.

Objective:To explore the incidence and risk factors of peripherally inserted central catheter (PICC) related upper extremity venous thrombosis (UEVT) in patients with head and neck neoplasm so as to provide a basis for preventing thrombosis.Methods:This study used the design of prospective cohort study. From January 2016 to March 2018, UEVT follow-up examination by B ultrasound was carried out for 1 137 head and neck neoplasm patients with PICC selected by convenience sampling. Single factor and multivariate Cox regression were used to determine the risk factors of PICC related UEVT.Results:There were 3.6% (41/1 137) of patients with PICC related UEVT. Multivariate Cox regression showed that the independent risk factors of PICC related UEVT included the older patients ( RR=1.04, 95% CI: 1.01-1.07, P=0.013) , being with a history of PICC catheterization ( RR=3.22, 95% CI: 1.53-6.77, P=0.002) and high frequency of catheter delivery ( RR=1.98, 95% CI: 1.30-3.00, P=0.001) . Conclusions:Patients with head and neck neoplasm have the low incidence of PICC related UEVT. The independent risk factors of PICC related UEVT in patients with head and neck neoplasm include the older ages, history of PICC catheterization and high frequency of catheter delivery. Positive intervention should be carried out for those patients which may reduce the incidence of PICC related thrombosis.

Objective: To facilitate using the CRISPR/Cas9 gene editing system in human liver and gallbladder cancer cells, we established Cas9 stably expressed human liver and gallbladder cancer cell lines, and validated the gene editing activity of Cas9. Methods: Human liver cancer cell lines (Huh7, PLC/PRF/5, HepG2, Hep3b, SK-HEP-1 and Li-7), human cholangiocarcinoma cells (RBE) and human gallbladder cancer cells (GBC-SD) were infected with 3 Cas9-expressing lentivirus vectors (pLv-EF1α-Cas9-Flag-Neo, pLv-EF1α-Cas9-Flag-Puro, Cas9m1.1), respectively, and Cas9 stably expressed colonies were screened and selected. We extracted the genomic DNA and protein, validated the stable expression of Cas9 by using genomic polymerase chain reaction (PCR) and western blot. Three of cell lines were further infected with Lv-EF1α-mCherry. Then mCherry positive cells were sorted by flow cytometry and infected with designed guide RNA (gRNA) vectors which targeted mCherry gene. Subsequently the gene editing activity of Cas9 was detected by genomic PCR, fluorescence microscopic observation and flow cytometry analysis. Results: One hundred Cas9-expressing human liver and gallbladder cancer cell lines were selected. Among them, 35 cell lines expressed Cas9-Neo, 25 expressed Cas9-puro, and 40 expressed mutant Cas9 (mCas9). We also established 3 cell lines with stable expression of mCherry (Huh7-mCas9-M, PLC/PRF/5-Cas9-M and SK-HEP-1-Cas9-M). The results of genomic PCR and sequencing showed that by lentiviral infection with 2 types of designed gRNA, the long fragment deletion of mCherry gene was found in these 3 cell lines. Moreover, mCherry^-EGFP⁺ cells infected with 2 types of gRNA were observed by fluorescence microscope. The results of flow cytometry showed that mCherry^-EGFP⁺ cells accounted from 0.3% to 93.6%. Conclusion We successfully establish 100 human liver and gallbladder cancer cell lines with stable expression of Cas9 protein and validate their activities of gene editing.