To establish an immunochromatographic method for rapid detection of caprine enterovir-us(CEV),the monoclonal antibody against CEV VP1 protein was used as gold-labeled monoclonal antibodies,and the purified rabbit-derived polyclonal antibody of CEV-VP1 and sheep anti-mouse IgG were used as the detection line and quality control line,respectively.The colloidal gold immu-nochromatographic test strips for CEV were prepared according to the principle of double antibody sandwich,evaluated,and applied for clinical specimen detection.The results showed that the meth-od specifically recognized CEV without cross-reaction with bovine enterovirus and bovine viral di-arrhea virus.The minimum detection limit of the method was 102.49 TCID50/mL and had good re-producibility.The prepared test strips had a shelf life of three months kept at 4 ℃.Detection of clin-ical samples using the immunochromatographic test strips showed 100％coincidence rate with RT-PCR method.In conclusion,the colloidal gold immunochromatographic test strips for detection of the emerging CEV with good specificity,sensitivity and repeatability,which provides a new techni-cal means easily used for the rapid detection/diagnosis and epidemiological investigation on CEV infection.

To establish an immunochromatographic method for rapid detection of caprine enterovir-us(CEV),the monoclonal antibody against CEV VP1 protein was used as gold-labeled monoclonal antibodies,and the purified rabbit-derived polyclonal antibody of CEV-VP1 and sheep anti-mouse IgG were used as the detection line and quality control line,respectively.The colloidal gold immu-nochromatographic test strips for CEV were prepared according to the principle of double antibody sandwich,evaluated,and applied for clinical specimen detection.The results showed that the meth-od specifically recognized CEV without cross-reaction with bovine enterovirus and bovine viral di-arrhea virus.The minimum detection limit of the method was 102.49 TCID50/mL and had good re-producibility.The prepared test strips had a shelf life of three months kept at 4 ℃.Detection of clin-ical samples using the immunochromatographic test strips showed 100％coincidence rate with RT-PCR method.In conclusion,the colloidal gold immunochromatographic test strips for detection of the emerging CEV with good specificity,sensitivity and repeatability,which provides a new techni-cal means easily used for the rapid detection/diagnosis and epidemiological investigation on CEV infection.

Tic disorder is a common neurodevelopmental disorder characterized by motor or vocal tics.The etiology and pathogenesis have not been fully elucidated.Increasingly, studies have shown that immune-mediated chronic inflammation of the nervous system plays an important role in the onset and progression of tic disorder.Histamine is an endogenous biogenic amine with peripheral and central immunomodulatory functions, which plays a crucial role in regulating the cortex-striatum-thalamus-cortex circuit, and exerts dual effects in the process of neuroinflammation.This review aims to investigate the pathological mechanism of histamine in the neuroinflammation of tic disorder.

Objective: To explore the relationship between visual acuity and physical fitness of university freshmen, so as to provide reference for myopia prevention and control for freshmen. Methods: From October to November 2022, 2 160 college freshman without majoring in public safety administration, selected from Guangxi Police College in 2022 by using the stratified cluster random sampling method, were reviewed for the results of visual acuity test and physical fitness scores. The physical fitness indices were evaluated by using the Z scores of physical fitness test scores, and the strength of association between the level of physical fitness index and myopia was analyzed by using Logistic regression model. Results: Among 2 160 college freshman without majoring in public safety administration, 917 (42.5%) students were diagnosed screening myopia, including 66 (3.1%) cases of high myopia, 383 (17.7%) cases of moderate myopia and 468 (21.7%) cases of mild myopia. The differences in the distribution of visual acuity tests among students with different physical fitness indices, body mass index, and gender were statistically significant ( Z/H=54.50, 49.53, 15.51, P <0.01). Low level and low middle level physical fitness indices were associated with screening myopia among freshmen[ OR (95% CI )=2.81(1.93-4.08),1.87(1.38-2.54)], and low level physical fitness indexes were associated with high myopia [ OR (95% CI )=7.22(2.33-22.32)] ( P <0.01). Conclusions Screening myopia among college freshman without majoring in public safety administration is related to physical fitness, and low level and low middle level physical fitness index are risk factors for myopia. Improving the level of physical fitness might be effective in preventing myopia.

OBJECTIVE To determine the contents of 15 components in Aurantii Fructus Immaturus from different origins （Citrus aurantium， C. junos， C. aurantium Linn.， C. sinensis Osb.， C. sinensis）， screen differential components， and provide references for the quality evaluation of Aurantii Fructus Immaturus. METHODS HPLC method was adopted to determine the contents of synephrine， N-methyltyramine， 5，7-dihydroxychromone-7-neohesperidoside， neoponcirin， narirutin， naringin， hesperidin， neohesperidin， naringenin， hesperetin， sinensetin， nobiletin， tangeretin， 5-demethylnobiletin， and auraptene in 46 batches of Aurantii Fructus Immaturus from different origins. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-0.1% formic acid （gradient elution） at the flow rate of 1.0 mL/min； column temperature was set at 40 ℃ ， detection wavelength was 284 nm， and sample injection volume was 5 μL. The differences between different origins of Aurantii Fructus Immaturus were analyzed by cluster analysis， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）， and differential components were screened. RESULTS The linear relationships of the aforementioned 15 components were all good within the tested mass concentration ranges （all r＞0.999 0）. The RSDs for precision， stability （24 h）， and repeatability tests were all less than 2.00%. The average recovery rate ranged from 91.1% to 103.9% （all RSDs＜3.10%）. Cluster analysis， PCA， and OPLS-DA revealed that C. sinensis Osb. and C. sinensis were clustered into one category， while C. aurantium，C. junos and C. aurantium Linn. were clustered into another category. The variable importance projection values for neohesperidin， auraptene， naringin， neoponcirin， tangeretin， hesperidin， sinensetin， and 5，7-dihydroxychromone-7-neohesperidoside were all greater than 1. CONCLUSIONS In this study， the contents of 15 components in Aurantii Fructus Immaturus from different origins are determined， and 8 differential components， including neohesperidin， auraptene， naringin， and neoponcirin， are screened out.

OBJECTIVE To evaluate the influence of pharmaceutical quality control on the efficiency and outcomes of standardized medication therapy management （MTM） services for patients with coronary heart disease by using Economic， Clinical and Humanistic Outcomes （ECHO） model. METHODS This study collected case data of coronary heart disease patients who received MTM services during January-March 2023 （pre-quality control implementation group， n＝96） and June-August 2023 （post-quality control implementation group， n＝164）. Using propensity score matching analysis， 80 patients were selected from each group. The study subsequently compared the economic， clinical， and humanistic outcome indicators of pharmaceutical services between the two matched groups. RESULTS There were no statistically significant differences in baseline data between the two groups after matching （P＞0.05）. Compared with pre-quality control implementation group， the daily treatment cost （16.26 yuan vs. 24.40 yuan， P＜0.001）， cost-effectiveness ratio [23.12 yuan/quality-adjusted life year （QALY） vs. 32.32 yuan/QALY， P＜0.001]， and the incidence of general adverse drug reactions （2.50% vs. 10.00%， P＝0.049） of post-quality control implementation group were decreased significantly； the utility value of the EuroQol Five-Dimensional Questionnaire （0.74± 0.06 vs. 0.71±0.07， P＝0.003）， the reduction in the number of medication related problems （1.0 vs. 0.5， P＜0.001）， the medication adherence score （[ 6.32±0.48） points vs. （6.10±0.37） points， P＝0.001]， and the satisfaction score （[ 92.56±1.52） points vs. （91.95±1.56） points， P＝0.013] all showed significant improvements. Neither group experienced serious adverse drug reactions. There was no statistically significant difference in the incidence of new adverse reactions between the two groups （1.25% vs. 3.75%， P＝0.310）. CONCLUSIONS Pharmaceutical quality control can improve the quality of pharmaceutical care， and the ECHO model can quantitatively evaluate the effect of MTM services， making pharmaceutical care better priced and more adaptable to social needs， thus being worthy of promotion.

OBJECTIVE: To explore the clinical phenotypes and genetic characteristics of a pedigree with Distal arthrogryposis type 5D (DA5D) caused by compound heterozygous variants in the ECEL1 gene. METHODS: A child (proband) diagnosed with DA5D and his family members (proband's parents and sister) who was admitted to the Department of Rehabilitation Medicine of Henan Children's Hospital in July 2022 due to "multiplex distal arthrogryposis" were enrolled into this study. Clinical data of the proband were collected and peripheral blood samples were obtained from the proband and members of his family about 3 mL. Trio-whole genome sequencing (trio-WGS) was carried out to detected the genetic variations of the proband and his family members. The candidate's pathogenic gene variants were screened and analyzed by Genome Aggregation Database (gnomAD) and other databases. The screened variants were annotated for clinical phenotypes using databases like the Online Mendelian Inheritance in Man (OMIM). The pathogenicity of the candidate variants was predicted by bioinformatics tools such as Provean. Based on the guidelines of the American College of Medical Genetics and Genomics (ACMG), pathogenicity ratings were conducted for variant sites. The protein conservation and mutation structure prediction of ECEL1 protein among species were carried out though MEGA-X and PyMOL. The research protocol of this study was reviewed by the Ethics Committee of Henan Provincial Children's Hospital (Approval No. 2023-H-H01), and informed consent for clinical research was obtained from the guardians of the probands. RESULTS: The proband had multiplex distal arthrogryposis involving hands, feet, knees, and ankles, and had right ptosis, micrognathia, low auricular position, and upturned nose. The parents and sister both had normal phenotypes. Trio-WGS and Sanger sequencing revealed that the child had compound heterozygous variants of paternal c.1742_c.1743insT and maternal c.2314T>G, for which the father and sister were carriers of the c.1742_c.1743insT heterozygous variant and the mother was carrier of c.2314T>A. Neither mutation site has been reported. According to guidelines of ACMG, the c.1742_c.1743insT variant was classified as likely pathogenic (PSV1+PM2_Supporting), and c.2314T>G was classified as uncertain (PM2_Supporting+PM3+PP3). The results of conserved analysis of amino acid residue sequences of ECEL1 protein showed that the missense mutation of the maternal c.2314T>G (p.Cys772Gly) was highly conserved among humans and other seven species. The protein structure prediction revealed that the c.1742_c.1743insT frameshift mutation led to the protein truncation, and the c.2314T>G missense mutation resulted in the failure of forming 1 disulfide bond. CONCLUSION The compound heterozygous variants of ECEL1 gene were considered to be pathogenic for this DA5D patient, which have expanded the mutational spectrum of the ECEL1 gene and provided a reference for clinical diagnosis as well as genetic counseling for this family.
Humans ; Pedigree ; Arthrogryposis/genetics* ; Male ; Female ; Heterozygote ; Phenotype ; Mutation ; Child ; Metalloendopeptidases

Trophoblast cells serve as the foundation for placental development. We analyzed published multiomics sequencing data and found that trophoblast cells highly expressed RRS1 compared to primitive endoderm and epiblast. We used HTR-8/SVneo cells for further investigation, and Western blot and immunofluorescence staining confirmed that HTR-8/SVneo cells highly expressed RRS1. RRS1 was successfully knocked down in HTR-8/SVneo cells using siRNA. Using IncuCyte S3 live-cell analysis system based on continuous live-cell imaging and real-time data, we observed that proliferation, migration, and invasion abilities were all significantly decreased in RRS1-knockdown cells. RNA-seq revealed that knockdown of RRS1 affected the gene transcription, and upregulated pathways in extracellular matrix organization, DNA damage response, and intrinsic apoptotic signaling, downregulated pathways in embryo implantation, trophoblast cell migration, and wound healing. Differentially expressed genes were enriched in diseases related to placental development. Consistent with these findings, human chorionic villus samples collected from spontaneous abortion cases exhibited significantly reduced RRS1 expression compared to normal controls. Our results highlight the functional importance of RRS1 in human trophoblasts and suggest that its deficiency contributes to early pregnancy loss.
Humans ; Trophoblasts/physiology* ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Female ; Pregnancy ; Abortion, Spontaneous/metabolism* ; Cell Line ; Placentation/genetics*

Objective·To observe the impact of hip synovitis on the long-term outcomes of free vascularized fibular grafting(FVFG)for osteonecrosis of femoral head(ONFH).Methods·Between October 2001 and December 2013,370 patients diagnosed with ONFH(556 hips)underwent FVFG.Preoperative synovitis was assessed using magnetic resonance imaging(MRI)and quantified with the Hip Inflammation MRI Scoring System(HIMRISS).Patients were divided into no synovitis group,moderate synovitis group,and severe synovitis group.Harris hip scores and the incidence of total hip arthroplasty were collected with an average follow-up duration of 90.5 months(range:5-215 months).Hip survival failure(defined as a Harris hip score lower than 80 at the final follow-up or the occurrence of total hip arthroplasty)was calculated.Multivariable Cox regression analysis was adopted to compare the influence of different degrees of synovial inflammation on long-term prognosis.Results·The proportion of hip survival failure was 28.0％in patients without synovitis and 28.5％in those with moderate synovitis,whereas it was significantly higher(60.4％)in patients with severe synovitis.The results of multivariable Cox regression analysis showed that severe synovitis was an independent risk factor for poor prognosis(HR 2.06,95％CI 1.21-3.53)after adjusting for age,gender,education level,marital status,ONFH type,affected side of ONFH,smoking history,baseline Harris hip score and other hip MRI-based covariates(collapse,bone marrow edema,and degeneration).Conclusion·Severe synovitis in patients with ONFH significantly increases the failure rate of hip preservation after FVFG,and the severity of synovitis should be considered in surgical decision-making.

Objective To investigate the detection rate of Klebsiella pneumoniae(KP)in the intestinal tract of intensive care unit(ICU)patients and community healthy adults,and analyze its virulence genes and drug resistance.Methods Fecal swabs or fecal specimens from ICU patients admitted to the Third Affiliated Hospital of Wenzhou Medical University and community healthy adults from December 2020 to December 2021 were collected for screening KP.The hypermucoviscosity(HM)phenotype was identified by the wire drawing test.The Vitek 2 automatic microbial analysis system and K-B disk diffusion method were used for detecting drug sus-ceptibility.PCR was used to screen extended spectrum β-lactamases(ESBLs),carbapenem resistance genes,9 virulence related genes,including allS,entB,fimH,iutA,kfu,magA,mrkD,rmpA,and ybtS,and 6 highly virulent capsule serotypes such as K1,K2,K5,K20,K54,and K57.Meanwhile,their multilocus sequence typing(MLST)was analyzed.Statistical analysis was conducted using SPSS 23.0 software.Results A total of 448 fecal samples were collected,including 140 from ICU patients and 308 from commu-nity healthy adults.A total of 89 strains of KP were isolated,including 36(25.7%)from ICU patients and 53(17.2%)from commu-nity healthy adults.The detection rate of KP in ICU patients was significantly higher than that in community healthy adults(χ2=4.375,P＜0.05).There were 9 strains(25%)of HMKP in ICU patients and 19 strains(35.8%)in community healthy adults,and there was no significant difference in the detection rate between them(χ2=1.170,P＞0.05).HMKP and KP detected in healthy adults were natu-rally resistant to ampicillin and still highly sensitive to commonly used antibiotics.The KP from ICU patients showed varying degrees of resistance to commonly used drugs.The resistance of the KP from ICU patients to cephalosporins,penicillinase inhibitors,carbapenem,quinolones,amtronam,gentamicin,amicacin and cotrimoxazole except for tobramycin was significantly higher than that from healthy a-dults(all P＜0.05).In addition,18 strains(50%)of KP producing ESBLs were detected,all of which were non-HM phenotypes.A-mong them,6 strains were carbapenem resistant KP(CRKP).Five strains of CRKP carrying KPC enzyme all belonged to ST11 type and 1 carrying NDM enzyme to ST1855 type.A total of 27(30.3%)strains of KP with highly virulent capsule serotypes were detected,mainly K1,K2,and K57,but no K5.There were no significant differences in the detection rates of 9 virulence genes and 5 highly vir-ulent capsule serotypes between ICU patients and healthy adults(all P＞0.05).High-risk ST types were detected in both ICU patients and healthy adults.Conclusion High virulence and high-risk KP strains are detected from the intestinal tract of healthy adults in this area,which increases the risk of infection transferring from community to hospital.In addition,carbapenemase-resistant ST11 is detec-ted from the intestinal tract of ICU patients,so the spread of drug-resistant plasmids should be highly vigilant.