OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

Objective:To assess the nutritional status of obese patients prior to bariatric surgery, and to explore the related factors of nutrient deficiency.Methods:Clinical data of 43 patients with obesity who underwent bariatric surgery at Beijing Hospital from Jan. 2011 to Dec. 2017 were retrospectively analyzed. Gender, age, BMI, body composition analysis data, blood test data of nutrients were collected. The software of SPSS 20.0 was used to conduct data analysis.Results:Nutrients deficiencies were found for vitamin D (100.0%), iron (26.1%), prealbumin (15.4%) and hemoglobin (7.0%). Hemoglobin, prealbumin, and serum iron levels were significantly higher in male patients than in female patients ( P=0.001, 0.000 and 0.001, respectively). Body fat percentage was negatively correlated with 25 hydroxyvitamin D ( r=-0.983, P=0.017), and positively correlated with serum sodium ( r=0.568, P<0.001). Conclusions:Obesity patients were presented with a variety of nutritional deficiencies before bariatric surgery. A comprehensive nutrients test should be performed before bariatric surgery, to detect and correct nutrient deficiencies preoperatively.

BACKGROUND: At the end of 2019, the outbreak of coronavirus disease 2019 (COVID-19) severely damaged and endangered people's lives. The public health emergency management system in China has played an essential role in handling the response to the outbreak, which has been appreciated by the World Health Organization and some countries. Hence, it is necessary to conduct an overall analysis of the development of the health emergency management system in China. This can provide a reference for scholars to aid in understanding the current situation and to reveal new research topics. METHODS: We collected 2247 international articles from the Web of Science database and 959 Chinese articles from the China National Knowledge Infrastructure database. Bibliometric and mapping knowledge domain analysis methods were used in this study for temporal distribution analysis, cooperation network analysis, and co-word network analysis. RESULTS: The first international article in this field was published in 1991, while the first Chinese article was published in 2005. The research institutions producing these studies mainly existed in universities and health organizations. Developed countries and European countries published the most articles overall, while eastern China published the most articles within China. There were 52 burst words for international articles published from 1999-2018 and 18 burst words for Chinese articles published from 2003-2018. International top-ranked articles according to the number of citations appeared in 2005, 2007, 2009, 2014, 2015, and 2016, while the corresponding Chinese articles appeared in 2003, 2004, 2009, and 2011. CONCLUSIONS There are differences in the regional and economic distribution of international and Chinese cooperation networks. International research is often related to timely issues mainly by focusing on emergency preparedness and monitoring of public health events, while China has focused on public health emergencies and their disposition. International research began on terrorism and bioterrorism, followed by disaster planning and emergency preparedness, epidemics, and infectious diseases. China considered severe acute respiratory syndrome as the starting research background and the legal system construction as the research starting point, which was followed by the mechanism, structure, system, and training abroad for public health emergency management.
Betacoronavirus ; China ; epidemiology ; Communicable Disease Control ; organization & administration ; Coronavirus Infections ; epidemiology ; prevention & control ; Disease Outbreaks ; prevention & control ; Humans ; Internationality ; Pandemics ; prevention & control ; Pneumonia, Viral ; epidemiology ; prevention & control

Objective To evaluate the clinical outcomes of hemodialysis patients after superficial femoral artery - superficial femoral vein arteriovenous graft (AVG). Methods Hemodialysis patients with mid - thigh superficial femoral artery - superficial femoral vein AVG from August 2015 to March 2018 in department of vascular surgery, Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine were enrolled. Their clinical outcomes and complications after operation were analyzed retrospectively. Patency rates were measured by Kaplan - Meier survival curve. Results A total of 18 cases were enrolled. The success rate of the operation was 100％without complication. Follow - up time was (22.00 ± 11.77) months with 100％ follow - up rate. The 6 months -, 12 months -, 24 months - primary patency rates were 83.3％±8.8％, 48.5％±12.1％, 24.2％± 13.5％, respectively; secondly patency rates were 100.0％, 100.0％, 87.5％±11.7％. There were 1 case of seroma, 1 case of puncture site infection, 11 cases of stenosis and 5 cases thrombosis during follow-up, while no heart failure, ischemia or pseudoaneurysm. Conclusion Mid - thigh AVG has low infection rate and high patency rate, so it can be as the first choice for the lower extremity AVG in hemodialysis patients.

Objective To summarize the experiences of aneurysmorrhaphy for arteriovenous fistula aneurysms with acute thrombosis in hemodialysis patients.Methods There were 7 cases of arteriovenous fistula with acute thrombosis from Nov 2015 to Feb 2017 at our department of vascular surgery,Longhua Hospital.Results In all cases thrombosis was secondary to proximal stenosis or occlusion.The stenosis and occlusion were corrected with embolectomy and aneurysmorrhaphy.The proximal part of the cephalic vein was translocated to the basilic vein in 1 case,axillary vein in 2 cases;autologous vein graft in 1 case;resection of the occlusion,end-to-end anastomosis in 1 case;autogenous patch in 1 case.No perioperative complications occurred.The operation site was cannulated within one month after operation in all cases.Patients were followed up for 7 months to 23 months,all cases were patent.Conclusions Aneurysmorrhaphy is effective,reliable and safe for arteriovenous fistula aneurysms with acute thrombosis in hemodialysis patients.

OBJECTIVETo use next generation sequencing (NGS) to identify unknown abnormality of chromosome 9 in a fetus and explore its mechanism.

METHODSA pregnant woman with abnormal fetal ultrasound finding underwent amniocentesis for G-banded chromosomal analysis. Karyotyping was also performed on peripheral blood samples derived from its parents. Fetal blood sample was obtained for NGS testing to identify abnormality unrecognized by karyotyping.

RESULTSAnalysis of amniocytes has revealed a 46,XX,der(9)(?::p21 to qter) karyotype, while both parents had a normal karyotype. NGS analysis of the fetus revealed a 20.67 Mb duplication (4 454 279-25 126 275) at 9p21.3p24.2, which overlapped with that of the 9p duplication syndrome, and a 4.43 Mb deletion at 9p24.2p24.3 (10 001-4 442 364), which partially overlapped with that of 9p deletion syndrome and 46,XY sex reversal 4 region. Comparison of the sequencing data with reference genome database indicated direct duplication of 9p21.3p24.2, which was also supported by review of the morphology of chromosome 9p. Therefore, the karyotype of the fetus was verified to be 46,XX,der(9) dir dup(9)(p21.3p24.2), del(9)(p24.2p24.3).

CONCLUSIONCombined G-banded karyotyping and NGS can identify dir dup del(9p) with accuracy. Delineation of the mechanism of dir dup del(9p) and its genotype-phenotype correlation may facilitate genetic counseling and estimation of recurrence risk.

Adult ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 9 ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Male ; Pregnancy ; Prenatal Diagnosis ; Trisomy ; genetics

OBJECTIVETo analyze a case with Angelman syndrome (AS) using single nucleotide polymorphism array (SNP array) and explore its genotype-phenotype correlation.

METHODSG-banded karyotyping and SNP array were performed on a child featuring congenital malformations, intellectual disability and developmental delay. Mendelian error checking based on the SNP information was used to delineate the parental origin of detected abnormality. Result of the SNP array was validated with fluorescence in situ hybridization (FISH).

RESULTSThe SNP array has detected a 6.053 Mb deletion at 15q11.2q13.1 (22,770,421- 28,823,722) which overlapped with the critical region of AS (type 1). The parents of the child showed no abnormal results for G-banded karyotyping, SNP array and FISH analysis, indicating a de novo origin of the deletion. Mendelian error checking based on the SNP information suggested that the 15q11.2q13.1 deletion was of maternal origin.

CONCLUSIONSNP array can accurately define the size, location and parental origin of chromosomal microdeletions, which may facilitate the diagnosis of AS due to 15q11q13 deletion and better understanding of its genotype-phenotype correlation.

Angelman Syndrome ; genetics ; Child ; Genotype ; Humans ; Karyotyping ; methods ; Male ; Phenotype ; Polymorphism, Single Nucleotide ; genetics

OBJECTIVETo investigate the prenatal application of single nucleotide polymorphism array (SNP array) in the identification of 5p deletion syndrome with partial trisomy 11q.

METHODSG-banded karyotyping and SNP array were performed using amniocytes on a fetus with multiple malformations for the identification of chromosome abnormality. Furthermore, karyotyping was carried out on the parental peripheral blood specimens to ascertain the origin of chromosome abnormalities and then fluorescence in situ hybridization (FISH) was also utilized to confirm the results.

RESULTSKaryotype of amniocyte showed 46, XY, der(5) (?::p15 → qter). SNP array revealed a 13.907 Mb deletion at 5p15.33p15.2 (chr5: 113576-14020561), overlapping the region of 5p deletion syndrome, and a 18.254 Mb duplication at 11q23.3 q25 (chr11: 116684627-134938470), overlapping no known syndrome. Karyotype of the parents showed a normal 46,XX in mother and 46,XY,t(5;11)(p15;q23) in father. Three-color metaphase FISH analysis on paternal peripheral blood specimens also confirmed the paternal karyotyping result.

CONCLUSIONSNP array could uncover 5p deletion syndrome with partial trisomy 11q unidentified by G-banded karyotyping and accurately locate the genomic breakpoints, facilitating the mapping of pathogenic critical regions and the identification of candidate genes, also accumulating research data for genotype-phenotype study.

Adult ; Chromosome Banding ; Chromosome Deletion ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Chromosomes, Human, Pair 11 ; genetics ; Chromosomes, Human, Pair 5 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis ; methods ; Trisomy ; diagnosis ; genetics

Objective To evaluate the role of c-Jun N-terminal kinase (JNK) signaling pathway in isoflurane (ISO) preconditioning against cerebral ischemia/reperfusion (I/R) injury and investigate the relationship between JNK signaling pathway and apoptosis.Methods Global cerebral I/R models were made by 4-artery occlusion technique.Forty male SD rats of clean grade were divided into sham operation group (S group),I/R injury group (I/R group),SP600125 (an inhibitor of JNK signaling pathway) + I/R group (SP + I/R group),ISO preconditioning group (ISO group),and ISO preconditioning + SP600125 group (ISO + SP group) according to the random number table.Preconditioning protocol was successive inhalation of 15 g/L ISO for 5 days,1 h/d.I/R was induced at 24 hours after the end of preconditioning.Brain tissues were harvested at 72 hours later to take histomorphological examination by HE staining as well as detect apoptosis of hippocampal nerve cells by TUNEL method,expression of caspase-3 in hippocampal nerve cells by immuno-histochemistry,and expression of protein p-JNK in hippocampal tissues by Western blot.Results Compared with S group,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 were significantly increased in the other groups (P ＜ 0.05).Moreover,p-JNK protein had a higher expression in IR group than in S group (P ＜ 0.05),but no significant difference was observed in SP + I/R group,ISO group,and ISO + SP group as compared with S group (P ＞ 0.05).Compared with I/R group,brain injury score,apoptosis ratio of nerve cells,expression of caspase-3,and expression of p-JNK protein were all declined in SP + I/R group,ISO group,and ISO + SP group (P ＜ 0.05).Moreover,brain injury score,apoptosis ratio of nerve cells,and expression of caspase-3 had further decline in ISO + SP group as compared with SP + I/R group and ISO group (P ＜ 0.05),but the difference in expression of p-JNK protein was insignificant among the three groups.Compared with SP + I/R group,no significant changes of each index were found in ISO group.Conclusion ISO preconditioning alleviates cerebral I/R injury through down-regulating expression of caspase-3 and inhibiting JNK signaling pathway.

OBJECTIVETo evaluate the therapeutic effects of transcatheter arterial chemoembolization (TACE) with or without radiofrequency ablation (RFA) in hepatocellular carcinoma (HCC) patients based on propensity score matching.

METHODSA logistic regression model was established with the treatment assignment as the dependent variable and the covariates as the independent variables. For each HCC patient, the propensity score was calculated from the model for caliper matching, and a survival analysis of the matched data were carried out.

RESULTSThe covariates between the groups were balanced after caliper matching based on the propensity scores. Before matching, the one-, two-, and three-year survival rates of TACE and TACE+RFA were 52.07% and 59.08%, 32.24% and 36.43%, and 316.54% and 19.39%, with the median survival time of 1.20 and 1.40 years, respectively, showing no significant differences in the overall survival rate between the two groups. After matching, the 1-year, 2-year, and 3-year survival rates of TACE and TACE+RFA groups were 54.39% and 62.28%, 23.15% and 40.08%, and 10.20% and 18.52%, with the median survival time of 1.10 years and 1.50 years, respectively, showing significant differences in the overall survival between the two groups. The survival rate in TACE+RFA group was higher than that of TACE only group.

CONCLUSIONPropensity score matching can effectively reduce the confounding bias of non-randomized clinical observational data for a more accurate evaluation of the therapeutic effect in HCC patients.

Adult ; Aged ; Carcinoma, Hepatocellular ; epidemiology ; mortality ; therapy ; Catheter Ablation ; Chemoembolization, Therapeutic ; Female ; Humans ; Liver Neoplasms ; epidemiology ; mortality ; therapy ; Logistic Models ; Male ; Middle Aged ; Propensity Score ; Survival Analysis ; Survival Rate