Objective To explore the clinical and genetic characteristics of children with holocarboxylase synthetase(HLCS)deficiency.Methods A retrospective analysis was conducted on the clinical data of 3 children with HLCS deficiency who were admitted to Children's Hospital Affiliated to Zhengzhou University from December 2014 to January 2024.Relevant literature indexed in CNKI,Wanfang Data,PubMed and other databases was reviewed to summarize the clinical characteristics and HLCS gene mutations of children with HLCS deficiency.Results All 3 children were male,with onset age of 4-6 months.The main clinical manifestations included shortness of breath,vomiting,diarrhea,and poor mental state,and partial cases were complicated by growth retardation and neurological symptoms.Laboratory tests showed metabolic acidosis in all cases,blood amino acid and acylcarnitine profiles as well as urinary organic acid analysis suggested multiple carboxylase deficiency.Genetic testing revealed compound heterozygous mutation in the HLCS gene of all 3 children,among which the c.1892delT(p.L631X)mutation was previously unreported.According to the guidelines of the American College of Medical Genetics and Genomics(ACMG),the c.1892delT(p.L631X)mutation was rated as pathogenic mutation(PVS1+PM2_supporting+PM3).Biotin supplementation was effective in all cases.Literature review included 27 English literatures and 29 Chinese literatures,reporting a total of 133 children with HLCS deficiency caused by HLCS gene mutation.Common clinical manifestations included metabolic acidosis,skin lesions,vomiting,feeding difficulties,dyspnea,diarrhea,and neurological symptoms,etc.Conclusions Blood amino acid and acylcarnitine profiles,urine organic acid analysis,and gene testing are helpful for the diagnosis of HLCS deficiency.Timely biotin supplementation leads to a good prognosis.The mutation of HLCS gene is considered as the genetic etiology of HLCS deficiency in 3 children,among which the c.1892delT(p.L631X)mutation is a newly discovered mutation.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective To investigate the risk factors for pneumonia complicating infectious mononucleosis(IM)in adults and construct a nomogram prediction model.Methods A retrospective analysis was conducted on 198 IM patients admitted to the Second Affiliated Hospital of Air Force Medical University from January 2015 to December 2021.Patients were divided into pneumonia group(n=52)and non-pneumonia group(n=146)based on whether pulmonary infection occurred during hospitalization.The baseline data(age,gender,place of onset,etc.),clinical manifestations(maximum body temperature,lymph node enlargement,splenomegaly,etc.),and inflammatory indicators[white blood cell count(WBC),C-reactive protein(CRP),etc.]were compared between the two groups.Kaplan-Meier curves were plotted to analyze the key indicators affecting the hospital stay of IM patients.Multivariate logistic regression was used to analyze the independent risk factors for pneumonia complicating IM in adults and construct a nomogram prediction model based on the identified risk factors.The predictive efficacy of the model was evaluated using the receiver operating characteristic(ROC)curve and the consistency of the model was assessed using the calibration curve.The fit of the model was evaluated using the Hosmer-Lemeshow test.Additionally,the sensitivity,specificity,and accuracy of the model were assessed using confusion matrix.Results Compared with non-pneumonia group,the pneumonia group had a significantly higher proportion of patients from rural areas,with body mass index(BMI)≥24 kg/m2,smoking history,hepatomegaly,fever duration of≥7 d,as well as increased total hospitalization costs and average daily hospitalization costs,and prolonged hospital stay(P<0.05).The proportion of patients with a history of antibiotic use was lower in the pneumonia group(P<0.05).Kaplan-Meier survival analysis showed that patients from rural areas,with BMI≥24 kg/m2,smoking history,no prophylactic use of antibiotics,fever duration≥7 d,and hepatomegaly had significantly prolonged hospital stays(P<0.05).Multivariate logistic regression analysis revealed that living in a rural area(OR=4.089,P<0.05),hepatomegaly(OR=4.082,P<0.05),and elevated WBC(OR=1.205,P<0.05)were independent risk factors for pneumonia complicating IM in adults,while the prophylactic use of antibiotics(OR=0.142,P<0.05)was an independent protective factor.The area under the ROC curve of the constructed nomogram prediction model was 0.827(95%CI 0.762-0.892),and the slope of the calibration curve was close to 1,and the Hosmer-Lemeshow test showed χ2=5.299,P=0.725,indicating good consistency and fit of the prediction model.The results of the confusion matrix assessment showed that the sensitivity of the model was 0.669(0.624-0.773),the specificity was 0.827(0.724-0.930),and the accuracy was 0.732(0.665-0.793).Conclusion The nomogram prediction model based on place of onset,hepatomegaly,the prophylactic use of antibiotics and WBC has excellent fit and discrimination,providing an effective quantitative tool for prognosis assessment of IM.

Arsenic is a semi-metal,and lipid-soluble arsenic compounds are one of the widespread forms in the environment and food chain,but there is a lack of standards for lipid-soluble arsenic compounds,which is one of the bottlenecks in the current analytical detection and toxicological studies of organic arsenic.In this study,four saturated arsenic-containing hydrocarbons,AsHC 318,AsHC 332,AsHC 346,and AsHC 374(The number is relative molecular mass),were successfully synthesized in three steps by using dimethylarsinic acid,potassium iodide,sodium hydroxide,and four brominated alkanes(1-Bromotetradecane,1-bromopentadecane,1-bromohexadecane,and 1-bromooctadecane)as raw materials.The structures of these four saturated arsenic-containing hydrocarbons were characterized by proton nuclear magnetic resonance(1H NMR)spectroscopy,13C nuclear magnetic resonance(13C NMR)spectroscopy,and high-resolution mass spectrometry(HR-MS).The yields of the method were 8%-10%,and the synthesized compounds could be used in subsequent toxicity evaluation experiments to assess the toxic effects and mechanisms of action of arsenic-containing hydrocarbons.This study provided an effective method for synthesis of arsenic-containing hydrocarbons,enriching the synthesis methods of arsenic-containing hydrocarbons,and provided raw materials for the subsequent toxicological studies of arsenic-containing hydrocarbons.

ObjectiveTo establish the multi-technique characteristic profiles of Alumen by X-ray diffraction（XRD）， Fourier-transform infrared spectroscopy（FTIR） and thermogravimetric-differential thermal analysis（TG-DTA）， and to explore the spectral characteristics for rapid identification of Alumen and its potential adulterant， Ammonium alum. MethodsA total of 27 batches of Alumen samples from 8 production regions were collected for preliminary identification based on visual characteristics. The PDF standard cards of XRD were used to differentiate Alumen from A. alum， and the XRD characteristic profiles of Alumen were established， and then the common peaks were screened. Based on hierarchical clustering analysis（HCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）， the characteristic information that could be used for identification of Alumen was selected with variable importance in the projection（VIP） value>1. FTIR characteristic profiles of Alumen were established， and key wavenumbers for identification were screened by HCA and OPLS-DA with VIP value>1. Meanwhile， the thermogravimetric differences between Alumen and A. alum were analyzed by TG-DTA， and the thermogravimetric traits that could be used for identification were screened. ResultsAlumen and A. alum could not be effectively distinguished by traits alone. However， by comparing the PDF standard cards of XRD， 15 batches of Alumen and 12 batches of A. alum could be distinguished. In the XRD profiles， 10 characteristic peaks were confirmed， corresponding to diffraction angles of 14.560°， 24.316°， 12.620°， 32.122°， 17.898°， 34.642°， 27.496°， 46.048°， 40.697° and 21.973°. In the FTIR profiles， 4 wavenumber ranges（399.193-403.050， 1 186.010-1 471.420， 1 801.190-2 620.790， 3 612.020-3 997.710 cm^-1） and 12 characteristic wavenumbers（1 428.994， 1 430.922， 1 432.851， 1 434.779， 1 436.708， 1 438.636， 1 440.565， 1 442.493， 1 444.422， 1 446.350， 1 448.279， 1 450.207 cm^-1） were identified. In the TG-DTA profiles， there were characteristic decomposition peaks of ammonium ion and mass reduction features near 555.34 ℃ for A. alum. These characteristics could serve as important criteria for distinguishing the authenticity of Alumen. ConclusionXRD， FTIR and TG-DTA can be used to rapidly detect Alumen and A. alum， and combined with the discriminant features selected through chemometrics， the rapid and accurate identification of Alumen and A. alum can be achieved. The research findings provide new approaches for the rapid identification of Alumen.

ObjectiveTo establish the multi-technique characteristic profiles of Alumen by X-ray diffraction（XRD）， Fourier-transform infrared spectroscopy（FTIR） and thermogravimetric-differential thermal analysis（TG-DTA）， and to explore the spectral characteristics for rapid identification of Alumen and its potential adulterant， Ammonium alum. MethodsA total of 27 batches of Alumen samples from 8 production regions were collected for preliminary identification based on visual characteristics. The PDF standard cards of XRD were used to differentiate Alumen from A. alum， and the XRD characteristic profiles of Alumen were established， and then the common peaks were screened. Based on hierarchical clustering analysis（HCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）， the characteristic information that could be used for identification of Alumen was selected with variable importance in the projection（VIP） value>1. FTIR characteristic profiles of Alumen were established， and key wavenumbers for identification were screened by HCA and OPLS-DA with VIP value>1. Meanwhile， the thermogravimetric differences between Alumen and A. alum were analyzed by TG-DTA， and the thermogravimetric traits that could be used for identification were screened. ResultsAlumen and A. alum could not be effectively distinguished by traits alone. However， by comparing the PDF standard cards of XRD， 15 batches of Alumen and 12 batches of A. alum could be distinguished. In the XRD profiles， 10 characteristic peaks were confirmed， corresponding to diffraction angles of 14.560°， 24.316°， 12.620°， 32.122°， 17.898°， 34.642°， 27.496°， 46.048°， 40.697° and 21.973°. In the FTIR profiles， 4 wavenumber ranges（399.193-403.050， 1 186.010-1 471.420， 1 801.190-2 620.790， 3 612.020-3 997.710 cm^-1） and 12 characteristic wavenumbers（1 428.994， 1 430.922， 1 432.851， 1 434.779， 1 436.708， 1 438.636， 1 440.565， 1 442.493， 1 444.422， 1 446.350， 1 448.279， 1 450.207 cm^-1） were identified. In the TG-DTA profiles， there were characteristic decomposition peaks of ammonium ion and mass reduction features near 555.34 ℃ for A. alum. These characteristics could serve as important criteria for distinguishing the authenticity of Alumen. ConclusionXRD， FTIR and TG-DTA can be used to rapidly detect Alumen and A. alum， and combined with the discriminant features selected through chemometrics， the rapid and accurate identification of Alumen and A. alum can be achieved. The research findings provide new approaches for the rapid identification of Alumen.

In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics.Similarities and variations of components present significant analytical challenges.A two-dimensional(2D)liquid chromatography-mass spectrometry(LC-MS)method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium(CMS).A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated.For efficient and high-accuracy screening of CMS,a targeted method based on a self-constructed high resolution(HR)mass spectrum database of CMS components was established.The database was built based on the commercial MassHunter Personal Compound Database and Library(PCDL)software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening.On this basis,the unknown peaks in the CMS chromatograms were deduced and assigned.The molecular formula,group composition,and origins of a total of 99 compounds,of which the combined area percentage accounted for more than 95％of CMS components,were deduced by this 2D-LC-MS method combined with the MassHunter PCDL.This profiling method was highly efficient and could distinguish hundreds of components within 3 h,providing reliable results for quality control of this kind of complex drugs.

Objective:To screen out the disinfection procedure and disinfectant suitable for the actual production of specific pathogen free(SPF)chicken embryo eggs,so as to ensure the disinfection effect of specific pathogen free(SPF)chicken embryo eggs in vaccine production.Methods:This study compares the microbial counting methods of soaking,swabbing with cotton swabs and pouring after thin-film filtration for SPF chicken embryo eggs in a GMP production workshop,and selects the most suitable method for SPF chicken embryo egg microbial counting.Experi-mental groups A,B,and C use self-prepared concentrations of 1∶50 sporicidal agent dilution solution,1∶128 alka-line phenol salt dilution solution,and ready-to-use compound quaternary ammonium disinfectant(sterile),respec-tively,and follow the actual disinfection procedures in the workshop to disinfect and sample the surfaces of SPF chicken embryo eggs entering different cleanliness grades,while using sterile water instead of disinfectant as the control group.The average bactericidal rate is calculated by recording the number of colonies and monitoring the viability of chicken embryo cells using microbial culture,and the disinfection effect of the three disinfectants on SPF chicken embryo eggs is evaluated.Results:The comparison of the results from the three methods shows that the method of sampling SPF chicken embryo eggs by immersion and then counting the microbial colonies through membrane filtration is superior to the other two methods.The final cleaning rate of the control group,which used sterilized injection water to clean the SPF chicken embryo eggs,was 91.67％to 96.97％,while the final steriliza-tion rate of the experimental group,which used the above three disinfectants to disinfect the SPF chicken embryo eggs,was 100.00％.By comparing the cell counts of the experimental group and the control group,it was found that the live cell density of the control group was(6.03-6.25)× 105 cells·mL-1,and that of the experimental groups A-C was(6.08-6.17)× 105 cells·mL-1,(5.99-6.25)× 105 cells·mL-1,and(5.87-6.21)× 105 cells·mL-1 respectively;the cell viability of the control group was 90.33％to 91.35％,and that of the experi-mental groups A-C was 88.25％to 92.12％,89.45％to 93.59％,and 88.02％to 92.89％respectively.Through statistical analysis,it was found that the P values of all experimental groups compared with the control group were greater than 0.05,indicating no statistically significant difference.Conclusion:By comparing the dis-infection effects,cell density and cell viability of the three disinfectants and comprehensively considering factors such as cost and risk of the three disinfectants,1∶50 sporicide dilution,1∶128 alkaline phenolate dilution and ready-to-use compound quaternary ammonium salt disinfectant(sterile)can all be used for the daily surface disin-fection of SPF chicken embryo eggs in the production workshop.The selection of an appropriate disinfectant should be based on specific application scenarios and requirements.

Objective To investigate the effect of LINC00839 on the malignant biological behavior of endometrial cancer cells by regulating the miR-625-5p/cytoplasmic polyadenylation element binding protein 4 (CPEB4) axis. Methods The cancer tissues and adjacent tissues of 46 patients with endometrial cancer were obtained,endometrial cancer cells of HEC-1B,Ishikawa and RL95-2 and human endometrial epithelial cells (HEEC) were cultured in vitro,and the expression levels of LINC00839,miR-625-5p and CPEB4 in tissues and cells were detected. HEC-1B cells were divided into the HEC-1B group (conventional culture),sh-Ctrl group (transfected with sh-Ctrl),sh-LINC00839 group (transfected with shRNA-LINC00839),anti-miR-625-5p group (transfected with shRNA-LINC00839 and miR-625-5p inhibitor),and anti-NC group (transfected with shRNA-LINC00839 and inhibitor-NC). The proliferation,apoptosis,migration and invasion abilities of HEC-1B cells in each group were compared,and the expressions of CPEB4 and epithelial-mesenchymal transition (EMT) related proteins were determined. The targeting relationships between LINC00839 and miR-625-5p,and miR-625-5p and CPEB4 were analyzed. Results The mRNA expression of LINC00839 and CPEB4,as well as the positive expression rate of CPEB4 protein in endometrial cancer tissues were higher than those in adjacent tissues,and the expression of miR-625-5p was lower than that in adjacent tissues (P<0.05). Compared with HEEC cells,the expression of LINC00839 and the mRNA and protein expression of CPEB4 in Ishikawa,RL95-2 and HEC-1B cells increased (P<0.05),and the expression of miR-625-5p decreased (P<0.05). Compared with the sh-LINC00839 group and anti-NC group,the protein expression of N-cadherin,Vimentin,and CPEB4,24-hour absorbance,and migration and invasion cell numbers of HEC-1B cells in the HEC-1B group,sh-Ctrl group and anti-miR-625-5p group increased (P<0.05),while the expression of E-cadherin protein and cell apoptosis rate decreased (P<0.05). There were binding sites between LINC00839 and miR-625-5p,miR-625-5p and CPEB4,with targeting regulatory relationships. Conclusion LINC00839 is related to the malignant biological behavior of endometrial cancer cells,interference with LINC00839 expression can inhibit the proliferation,invasion and migration,and promote apoptosis of HEC-1B cells,and its mechanism may be achieved by regulating the miR-625-5p/CPEB4 axis.

Aim To explore the uric acid lowering effects and mechanisms of acteoside(AS)and echina-coside(ECH)extracted from Cistanche tubulosa.Methods Hyperuricemia(HUA)rat model was es-tablished using potassium oxonate(1.5 g·kg-1·d-1)combined with adenine(100 mg·kg-1·d-1).The HUA model rats were randomly divided into a model control group,an allopurinol group(30 mg·kg-1·d-1),AS low and high(50,100 mg·kg-1·d-1)dose groups,and ECH low and high(50,100 mg·kg-1·d-1)dose groups.A normal control group was previously established.After four weeks of admin-istration,the blood biochemistry analyzer was used to detect SUA UREA,CREA,adenosine deaminase(ADA)and uric acid(UUA)levels in urine.Enzyme linked immunosorbent assay was used to detect xan-thine oxidase(XOD)levels in serum.Kidney tissue was collected,and kidney coefficient was weighed and calculated.Hematoxylin eosin(HE)staining was used to observe renal pathological changes.Immunohisto-chemistry(IHC)and Real time PCR were used to de-tect the protein and mRNA expression levels of urate transporter 1(UR AT1),glucose transporter 9(GLUT9),organic anion transporter 1(OAT1),and organic anion transporter 3(OAT3)in renal tissue.Results AS and ECH could improve renal function in HUA model rats,reduce SUA levels,promote uric acid excretion,downregulate serum levels of ADA and XOD,and upregulate protein and mRNA expression of URAT1 and GLUT9 in renal tissue,while upregulating protein and mRNA expression of OAT1 and OAT3 in renal tissue.Conclusions The phenylethanoid glyco-sides AS and ECH from Cistanche tubulosa may exert uric acid lowering effects by improving renal function in HUA model rats,reducing ADA and XOD levels,and regulating the expression of URAT1,GLUT9,OAT1,and OAT3.