AIM: To conduct whole exome sequencing(WES)analysis on three pedigrees with blepharophimosis-ptosis-epicanthus inversus syndrome(BPES)to identify the pathogenic gene loci, uncover novel mutations, and expand the mutation spectrum of the disease-associated genes.METHODS:Retrospective study. A total of 3 pedigrees and 30 patients with BPES(with criteria of bilateral blepharophimosis, ptosis, epicanthus inversus and wider inner canthal distance at birth)treated in the Ophthalmology Department of the Second People's Hospital of Yunnan Province were collected from January 2021 to August 2021, including 8 patients and 22 unaffected family members. Peripheral blood samples were collected from patients and related family members, and genomic DNA was extracted for whole exome sequencing. The sequencing results were screened to identify potential pathogenic gene loci, and candidate mutations were validated using Sanger sequencing.RESULTS:WES analysis identified pathogenic gene mutations in 3 BPES pedigrees: pedigree 1(6 members, 3 affected individuals, with a history of disease across three generations)harbored a novel heterozygous mutation in the PIEZO2 gene(located 36 bp upstream of exon 11, G>C). Sanger sequencing confirmed that this mutation was present in all affected individuals and absent in normal family members, and it represents the first report of this mutation. Pedigree 2(14 members, 2 affected individuals)and pedigree 3(10 members, 3 affected individuals)carried known heterozygous mutations in the FOXL2 gene, namely the missense mutation c.313A>C(p.N105H)and the in-frame mutation c.672_701dupAGCGGCTGCAGCAGCTGCGGCTGCAGCCGC(p.A225_A234dupAAAAAAAAAA), respectively.CONCLUSION:WES successfully identified the pathogenesis of familial congenital BPES in two families, including a known FOXL2 gene mutation and a newly discovered PIEZO2 gene mutation. These findings provide a theoretical basis for genetic counseling and reproductive guidance. Notably, the PIEZO2 gene mutation(located 36 bp upstream of exon 11, G>C)discovered in the pedigree 1 is reported for the first time and plays a critical role in the onset of the disease in this family. Further investigation of this new mutation could not only expand the mutation spectrum of BPES, but also enhance our understanding of its pathogenic mechanisms.

摘要 目的 探讨血浆神经胶质细胞原纤维酸性蛋白（GFAP）及同型半胱氨酸（Hcy）水平对肝豆状核变性（又称肝豆状核变性，WD）的诊断及肝脑型鉴别诊断价值。方法 招募安徽中医药大学第一附属医院脑病中心2023年1月—2025年1月收治的WD患者共120例，其中WD脑型63例，WD肝型57例，以及同期在体检中心筛查的30名健康志愿者。采用ELISA法测定纳入对象血浆GFAP、Hcy水平，进行组间差异性比较和ROC曲线分析。并运用Spearman相关分析探讨血浆GFAP、Hcy水平与统一肝豆状核变性评定量表评分（UWDRS）、24 h尿铜及血清铜蓝蛋白（CER）水平的相关性。结果 WD肝型及脑型患者的血浆GFAP水平均显著高于对照组（P<0.05），且WD脑型较WD肝型升高更为显著（P<0.05）。血浆Hcy水平也在WD肝型及脑型患者中明显升高（P<0.05），但在WD肝型、脑型间未表现出显著差异。血浆GFAP诊断WD脑型的曲线下面积（AUC）为0.861，截断值为135.71 pg/ml，敏感度68.3％，特异度82.3％；该指标诊断WD肝型的AUC为0.695，截断值为129.84 pg/ml，敏感度64.7％，特异度83.3％；其在WD肝型及脑型鉴别诊断中的AUC为0.75，截断值为151.12 pg/ml，敏感度73.9％，特异度87.8％。血浆Hcy诊断WD的AUC为0.788，截断值为15.59 μmol/L，敏感性77.9％，特异性66.7％。Spearman相关性分析结果显示，WD肝型及脑型患者血浆GFAP、Hcy水平与UWDRS评分及24 h尿铜水平均呈正相关（P<0.05），与CER水平无显著相关性（P>0.05）。结论 血浆GFAP、Hcy水平与WD神经及肝脏功能受损程度密切相关，且为早期诊断WD以及血浆GFAP对WD各分型的鉴别诊断提供了一定的临床价值。 Abstract Objective To investigate the value of plasma glial fibrillary acidic protein （GFAP） and homocysteine （Hcy） in the diagnosis of hepatolenticular degeneration （also know as Wilson disease， WD） and the differential diagnosis of the hepatic and neurological forms of WD. Methods A total of 120 WD patients who were admitted to Encephalopathy Center of our hospital from January 2023 to January 2025 were enrolled， among whom there were 63 patients with neurological WD and 57 patients with hepatic WD， and 30 healthy volunteers who underwent physical examination during the same period of time were enrolled as control group. ELISA was used to measure the plasma levels of GFAP and Hcy， and the differences between groups were analyzed. The receiver operating characteristic（ROC） curve analysis was performed， and the Spearman correlation analysis was used to investigate the correlation of the plasma levels of GFAP and Hcy with Unified Wilson Disease Rating Scale （UWDRS） score， 24-hour urinary copper， and the serum level of ceruloplasmin （CER）. Results The patients with hepatic or neurological WD had a significantly higher plasma level of GFAP than the control group（P<0.05）， and the patients with neurological WD had a significantly greater increase than those with hepatic WD（P<0.05）. The patients with hepatic or neurological WD also had a significant increase in the plasma level of Hcy（P<0.05）， but with no significant difference between the patients with hepatic WD and those with neurological WD.Plasma GFAP had an area under the ROC curve（AUC） of 0.861 in the diagnosis of neurological WD， with a cut-off value of 135.71 pg/ml， a sensitivity of 68.3%，and a specificity of 82.3%；plasma GFAP had an AUC of 0.695 in the diagnosis of hepatic WD， with a cut-off value of 129.84 pg/ml， a sensitivity of 64.7%， and a specificity of 83.3%； in the differential diagnosis of hepatic and neurological WD， plasma GFAP had an AUC of 0.75， with a cut-off value of 151.12 pg/ml，a sensitivity of 73.9%， and a specificity of 87.8%. Plasma Hcy had an AUC of 0.788 in the diagnosis of WD， with a cut-off value of 15.59 μmol/L， sensitivity of 77.9%， and specificity of 66.7%. The Spearman correlation analysis showed that in the patients with hepatic or neurological WD， the plasma levels of GFAP and Hcy were positively correlated with UWDRS score and 24-hour urinary copper （P<0.05）， but they were not significantly correlated with the level of CER （P>0.05）. Conclusion The plasma levels of GFAP and Hcy are closely associated with the degree of neurological and hepatic impairment in WD， which provides a certain clinical value for the early diagnosis of WD and the differential diagnosis of hepatic and neurological WD.
Homocysteine

BACKGROUND:Intestinal flora is a diverse and dynamic microbial community located in the human gastrointestinal tract,which plays a crucial role in maintaining human immunity and health.In recent years,the concept of"gut-lung axis"has been proposed,suggesting that intestinal flora is closely related to the lung,and exercise can regulate respiratory diseases by maintaining the balance of intestinal flora.OBJECTIVE:To review the relationship between intestinal flora and different respiratory diseases such as pneumonia,lung cancer,asthma and chronic pulmonary obstruction,and the influence of different exercise modes on intestinal flora and respiratory diseases,providing new ideas for an in-depth exploration of the mechanisms by which exercise regulates the role of intestinal flora in respiratory diseases.METHODS:CNKI and PubMed databases were searched for relevant literature published from 1944 to 2024,using the search terms of"intestinal flora,gut bacteria,respiratory illness,pneumonia,lung cancer,asthma,chronic obstructive pulmonary diseases,aerobic exercise,resistance training"in Chinese and English,respectively.Totally 101 documents were included according to inclusion and exclusion criteria.RESULTS AND CONCLUSION:(1)Intestinal flora plays an important role in the regulation of respiratory diseases such as pneumonia,lung cancer,asthma and chronic pulmonary obstruction diseases.(2)Different exercise modes are closely related to intestinal flora.Aerobic exercise can play a beneficial role in the regulation of intestinal flora by improving insulin sensitivity,increasing microbial diversity and inhibiting systemic chronic inflammation.Resistance exercise can decrease the permeability of intestinal mucosa and promote the production of short-chain fatty acids.Aerobic exercise combined with resistance exercise can also increase the diversity of intestinal flora and affect the composition of intestinal flora.(3)Exercise can improve respiratory diseases by regulating inflammatory response and reducing oxidative stress damage as well as improving cardiopulmonary function and exercise performance.(4)Exercise plays a key role in the prevention and treatment of respiratory diseases by regulating intestinal flora,inhibiting inflammatory response,regulating oxidative stress,improving intestinal barrier permeability and maintaining intestinal flora homeostasis.

Objective To observe the origin and course of the obturator artery(OA),so as to provide anatomical reference for reducing hemorrhage during pelvic surgery and pubic fracture fixation.Methods A total of 65 human hemi-pelvises specimens with intact structure were dissected to observe the origin,course and other variations of OA.Measure the length of the inner section of OA basin and the outer diameter at the origin,etc.Results OA originated from the internal iliac artery in 57 cases(87.7%),including 3 cases(4.6%)of the superior gluteal artery,5 cases(7.7%)of the inferior gluteal artery,3 cases(4.6%)of the external iliac artery and 5 cases(7.7%)of the inferior epigastric artery.OA participated in the formation of the arterial trunk in 3 cases(4.6%).The length of the pelvic segment of the OA in male and female was(50.87±15.41)mm and(51.71±14.19)mm,respectively,with no statistically significant difference between them(P>0.05).The outer diameters at the origin of the OA in male and female were(2.79±1.05)mm and(2.35±0.86)mm,and there was no statistically significant difference between them(P>0.05).Conclusion OA mainly originated from the anterior trunk of the internal iliac artery,with a few OA originated from the branches of the posterior trunk or the inferior epigastric artery,or participated in the formation of the arterial trunk.In pelvic surgery involving OA area,attention should be paid to the length of its pelvic segment and the outer diameter at the origin of OA,so as to better locate and protect blood vessels during surgery.

Colorectal cancer is one of the common malignancies of the digestive system.Laparo-scopic surgery,as a minimally invasive procedure,has advantages such as minimal trauma,rapid re-covery,and shortened hospital stay,gradually becoming a main choice for treatment of digestive system diseases.However,anastomotic leakage remains one of the serious complications following this surger-y,impacting patients'recovery and prognosis.This review aimed to summarize and analyze recent re-search on anastomotic leakage after laparoscopic colorectal cancer surgery,exploring its pathogenesis,risk factors,preventive strategies,treatment and nursing measures,in order to provide a scientific ba-sis and guidance for clinical practice.

Objective:To investigate the influence of ultrasonic monitoring of bladder filling levels on setup errors before fractionated radiotherapy for cervical cancer patients through a comparative analysis, and its effectiveness in improving clinical target volume (CTV) margins.Methods:A retrospective study was conducted on 1 284 error data of setup via cone beam CT (CBCT) and 6D setup error correction system from 172 cervical cancer patients treated in the Radiotherapy Department of Peking University Cancer Hospital from January 2019 to October 2023. These patients were classified into two groups: 87 (659 times of setup) with ultrasonic monitoring of bladder filling levels and 85 (625 times of setup) without ultrasonic monitoring. The setup errors, error distributions, and numbers of abnormal setups between the two groups were compared in the lateral (Lat), longitudinal (Lng), vertical (Vrt), pitch (Pitch), roll (Roll), and rotational (Rtn) dimensions. Moreover, the CTV to planning target volume(PTV) margin values in the three-dimensional direction were calculated for both groups to assess the clinical value of ultrasonic monitoring of bladder filling levels before fractionated radiotherapy.Results:Compared to the group without ultrasonic monitoring, the group with ultrasonic monitoring exhibited lower median values of setup errors in all six-dimensional directions and smaller upper and lower interquartile ranges ( Z = -10.86 to -6.34, P<0.05). The group with ultrasonic monitoring manifested more concentrated setup errors in various directions and statistically significantly reduced numbers of abnormal setups ( χ2=15.33, P<0.05). Moreover, CTV-PTV margins of the group with ultrasonic monitoring displayed reduced CTV-PTV margin values by 0.55, 1.52, and 1.26 mm in the Vrt, Lng, and Lat directions, respectively. Conclusions:Pre-radiotherapy ultrasonic monitoring of bladder filling levels in cervical cancer patients can significantly improve the repeatability of setup, thus notably reducing the incidence of abnormal setups. Theoretically, it can narrow the range from the CTV to the PTV, thereby minimizing radiation exposure to healthy tissues and ultimately enhancing radiotherapy precision for cervical cancer and reducing radiation damage.

OBJECTIVE T o explore the value of targeted next-generation sequencing(t-NGS)in diagnosis of respir-atory tract pathogens through meta-analysis so as to provide reference for clinical application.METHODS PubMed database,Web of Science database,Wanfang database,CNKI database and Sinomed database were retrieved,and the time period of retrieval ranged from Jan.2010 to May 2024.The literatures were screened out based on the es-tablished standards.The quality was assessed by QU ADAS-2,the risk of bias graph was drawn by Revman 5.4,and the statistical analysis was performed by Stata 16.0.RESULTS A total of 9 literatures were included in the study.The result of meta-analysis showed that the heterogeneity test Q for sensitivity was 268.21,P＜0.01,I2=97.02％,with the heterogeneity test Q for specificity 210.04,P＜0.01,I2=96.19％,the combined sensitivity 0.88(95％CI:0.62 to 0.97),combined specificity 0.68(95％CI:0.41 to 0.86),combined positive likelihood ratio 2.72(95％CI:1.44 to 5.15),combined negative likelihood ratio 0.18(95％CI:0.06 to 0.53),combined diagno-sis score 2.74(95％CI:1.68 to 3.80),and combined diagnostic odds ratio 15.44(95％CI:5.34 to 44.66).The area under synthesize receiver operating characteristic(SROC)curve(AUC)was 0.85(95％CI:0.82 to 0.88).The result of Deeks funnel plot showed that P was 0.99,indicating that there was no obvious publication bias.CONCLUSIONS The sensitivity of tNGS is high in detection of the pathogens causing acute respiratory tract infection,the specificity needs to be improved,but its comprehensive ability is satisfactory.It has certain val-ue in early clinical diagnosis.

BACKGROUND: Changes in N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels may not fully translate into patient-reported health status in patients with heart failure (HF). We aimed to evaluate the correlation between NT-proBNP levels and patient-reported health status changes at one month after discharge of patients, and their associations with risk of death and rehospitalization in patients with acute HF. METHODS: We used data from the China Patient-centered Evaluative Assessment of Cardiac Events Prospective Heart Failure Study (PEACE 5p-HF Study). Patient-reported health status was measured by the 12-item Kansas City Cardiomyopathy Questionnaire (KCCQ-12). Patients who were hospitalized for HF and completed the KCCQ-12 and NT-proBNP tests before and one month after discharge were eligible in our study. We stratified patients into different groups based on NT-proBNP levels (i.e., improved, stable, and deteriorated) and KCCQ-12 scores (i.e., not deteriorated and deteriorated). We also examined the associations of the joint NT-proBNP and KCCQ-12 change with the risk of one-year and four-year clinical outcomes. RESULTS: A total of 2461 patients were included in the analysis. The mean age was 64.06 ± 13.51 years, and 36.37% (895/2461) of the study population were female. Among patients with improved NT-proBNP levels, 115 (10.95%) patients had deteriorated KCCQ-12 scores. The correlation between the change in the KCCQ-12 score and NT-proBNP level was weak ( r2 = 0.002, P = 0.013). Stratification by changes in the KCCQ-12 score revealed subgroups with distinctive risks, such that patients with deteriorated KCCQ-12 scores in any of the NT-proBNP change groups exhibited an increased risk of one-year all-cause death than participants with not deteriorated KCCQ-12 scores in any of the NT-proBNP change groups. Patients with improved NT-proBNP levels and deteriorated KCCQ-12 scores presented greater risks of one-year all-cause death (hazard ratio [HR]: 2.45, 95% confidence interval [CI]: 1.34-4.48) than patients with stable NT-proBNP levels and not deteriorated KCCQ-12 scores (HR [95% CI], 1.77 [1.25-2.53]). CONCLUSIONS: A discrepancy between changes in NT-proBNP levels and KCCQ-12 scores was common. The change in NT-proBNP levels was not sufficient to characterize critical aspects related to HF during one month after discharge of patients. Changes in the KCCQ-12 score exhibit complementary information to NT-proBNP levels for the prediction of clinical outcomes in patients with acute HF. REGISTRATION www.clinicaltrials.gov (No. NCT02878811).
Aged ; Female ; Humans ; Male ; Middle Aged ; Health Status ; Heart Failure/metabolism* ; Natriuretic Peptide, Brain/metabolism* ; Peptide Fragments/metabolism* ; Prospective Studies

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

OBJECTIVE T o explore the value of targeted next-generation sequencing(t-NGS)in diagnosis of respir-atory tract pathogens through meta-analysis so as to provide reference for clinical application.METHODS PubMed database,Web of Science database,Wanfang database,CNKI database and Sinomed database were retrieved,and the time period of retrieval ranged from Jan.2010 to May 2024.The literatures were screened out based on the es-tablished standards.The quality was assessed by QU ADAS-2,the risk of bias graph was drawn by Revman 5.4,and the statistical analysis was performed by Stata 16.0.RESULTS A total of 9 literatures were included in the study.The result of meta-analysis showed that the heterogeneity test Q for sensitivity was 268.21,P＜0.01,I2=97.02％,with the heterogeneity test Q for specificity 210.04,P＜0.01,I2=96.19％,the combined sensitivity 0.88(95％CI:0.62 to 0.97),combined specificity 0.68(95％CI:0.41 to 0.86),combined positive likelihood ratio 2.72(95％CI:1.44 to 5.15),combined negative likelihood ratio 0.18(95％CI:0.06 to 0.53),combined diagno-sis score 2.74(95％CI:1.68 to 3.80),and combined diagnostic odds ratio 15.44(95％CI:5.34 to 44.66).The area under synthesize receiver operating characteristic(SROC)curve(AUC)was 0.85(95％CI:0.82 to 0.88).The result of Deeks funnel plot showed that P was 0.99,indicating that there was no obvious publication bias.CONCLUSIONS The sensitivity of tNGS is high in detection of the pathogens causing acute respiratory tract infection,the specificity needs to be improved,but its comprehensive ability is satisfactory.It has certain val-ue in early clinical diagnosis.