Objective: To construct gene recombinant expression plasmids of human anti-P antibody with IgG1 and kappa chain based on the human hybridoma cell line. Methods: Starting from the specific RT-PCR products encoding the variable regions of the IgH chain and IgL chain of the anti-P monoclonal cell line, appropriate restriction enzyme digestion sites were introduced at both ends of the VH and VL fragments through nested PCR. The plasmids carrying the antibody constant region and the nested PCR products of VH and VL were ligated by the action of T4 ligase and subsequently transferred into competent E. coli DH5ɑ, and positive clones were selected in the antibiotic resistant LB medium. After sequence confirmation, recombinant plasmids DNA were transfected into HEK293T cells, and the recombinant antibody obtained in the culture supernatant. The characteristics of recombinant expression antibodies were determined by using rapid antibody isotying kit, serological agglutination tests and flow cytometry. Results: Recombinant gene expression vectors, pFUSEss-CHIg-hG1+VH, pFUSE2ss-CLIg-hκ+VL, were successfully constructed. Human IgG1 kappa light chain antibodies were detected in the supernatant of HEK293T cells transient transfected with recombinant plasmids. After the supernatant was ultra-filtered and concentrated, it could cause agglutination reactions with P antigen-positive red blood cells. The mean fluorescence intensity (MFI) of the reaction between recombinant antibodies and antigen-positive red blood cells in flow cytometry experiments was higher than that of antigen-negative red blood cells. Conclusion: The experimental study on the conversion of red blood group antibody types by genetic engineering technology represents a beneficial exploration towards establishing a feasible technical route for the development of genetic recombination and modification of antibodies reagent.

Objective:To analyze the variation trends in the levels of apolipoprotein B (ApoB), the ratio of apolipoprotein B to low-density lipoprotein cholesterol (ApoB/LDL-C), and non-high density lipoprotein cholesterol (non-HDL-C) among patients and individuals undergoing physical examinations at Zhongshan Hospital, Fudan University from 2014 to 2024.Methods:Data on ApoB, LDL-C, and nonHDL-C levels were collected from individuals (the overall population) who visited or underwent physical examinations at our hospital from January 1, 2014 to December 31, 2024, as well as from individuals (the fixed population) who were tested annually over the 11-year period. The Mann-Kendall test combined with Sen slope estimation was used to analyze the trends. The abnormal rates of ApoB and non-HDL-C in both the overall and fixed populations, as well as the changes in the first and last test results over the 11 years in the fixed population, were also analyzed.Results:The overall population totaled 1 679 440 cases, aged 57 (45, 67) years, and 718 738 cases (42.8%) were female, of which 1 250 234 cases (74.4%) were in the patient population and 429 206 cases (25.6%) were in the health check population. The fixed population of 1 560 cases was 56 (45, 65) years old and 655 cases (42.0%) were female, of which 1 044 cases (66.9%) were in the patient population and 516 cases (33.1%) in the health check population. Between 2014 and 2024, the overall population ApoB decreased from 0.842 g/L to 0.822 g/L (Sen slope -0.001 g·L -1·year -1, P=0.01), the abnormality rate decreased from 24.70% to 22.03% (Sen slope -0.50%/year, P=0.06), and 69.4% of the fixed population showed no change in the subgroups of initial and final tests. ApoB/LDL-C increased from 0.824 to 0.868 (Sen slope -0.003/year, P=0.72). Non-HDL-C decreased from 3.46 mmol/L to 3.32 mmol/L (Sen slope 0.018 mmol·L -1·year -1, P=0.35), with a statistically significant upward trend since 2019 (Sen slope 0.037 mmol·L -1·year -1, P=0.008), and the anomaly rate decreased from 24.3% to 22.7% (Sen slope 0.55%/year, P=0.64), with a statistically significant upward trend since 2019 (Sen slope 1.53%/year, P=0.014), and no change in the grouping of the first and last tests in 75.0% of the fixed population. The trends in the above items were consistent between the overall population and the fixed population. 20-<40 year olds had the most significant upward trend in non-HDL-C along with the least significant downward trend in ApoB/LDL-C and ApoB. Conclusion:ApoB in the overall and fixed populations from 2014-2024 in a Shanghai hospital showed a decreasing trend, and non-HDL-C and abnormality rates showed an increasing trend from 2019 onwards.

Objective:To analyze the trend of lipid changes in patients and individuals undergoing physical examination at Zhongshan Hospital of Fudan University from 2014 to 2024, providing evidence for the formulation of cardiovascular disease prevention and control strategies.Methods:A total of 2 657 835 individuals (general population) who underwent lipid testing during medical visits or physical examinations at Zhongshan Hospital of Fudan University from January 1, 2014, to December 31, 2024, were selected. Among them, 6 234 individuals who were tested consecutively for 11 years were considered as the fixed population. Lipid levels were analyzed across different genders and age groups. Total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were analyzed. The trends in lipid changes and the abnormal rates of TC (≥5.18 mmol/L) and LDL-C (≥3.40 mmol/L) in both the general and fixed populations were statistically analyzed.Results:The median age of the general population was 53 (41, 63) years, with 1 498 533 males (56.4%); 1 149 662 individuals (43.3%) were from the physical examination group. The median age of the fixed population was 52 (39, 62) years, with 3 262 males (52.3%); 2 955 individuals (47.4%) were from the physical examination group. Over an 11-year period, the logarithmically transformed TG (lnTG) in the general population slightly increased from 1.35 mmol/L to 1.36 mmol/L (Sen slope=0.007 mmol·L -1·year -1; S=27, P=0.043). Although there were fluctuations in TC, LDL-C, and HDL-C, the trends were not statistically significant ( P>0.05). However, in the subset of the population undergoing regular health check-ups, TC showed a steady increase over time ( S=27, P=0.043). Within a fixed population over the same 11-year period, there were no statistically significant changes in lipid profiles ( P>0.05). Nevertheless, in the fixed subset undergoing regular health check-ups, both TC and lnTG exhibited an upward trend (TC: S=27, P=0.043; lnTG: S=31, P=0.020), while in the fixed subset seeking medical attention, TC and LDL-C demonstrated a downward trend (TC: S=-31, P=0.020; LDL-C: S=-27, P=0.043). Trends in lipid profiles varied among different genders and age groups. Specifically, both men and women aged 20-<40 years old showed an increase in TC, abnormal TC rates, and abnormal LDL-C rates ( P<0.05). Conversely, in the fixed population, women over 60 years old exhibited a decrease in TC, abnormal TC rates, and abnormal LDL-C rates ( P<0.05). Conclusion:During the period from 2014 to 2024, there were slight fluctuations in the average lipid levels of both the general and fixed populations. Notably, TC, abnormal TC rates, and abnormal LDL-C rates increased among men and women aged 20-<40 years old, while these parameters decreased among women over 60 years old in the fixed population.

Synovium is the target organ of rheumatoid arthritis.The excessive proliferation of synovial cells and insufficient apoptosis lead to synovial hyperplasia,which in turn causes damage to the surrounding tissues of the joint and bone destruction."Yang transforming qi and yin forming elements"is derived from Su Wen and is a highly summarized description of the functions of yin and yang,which runs through the entire course of the disease.This article elucidated the theoretical connotation of"yang transforming qi and yin forming elements"and its connection with synovial hyperplasia,proposing that the insufficiency of"yang transforming qi"is the root of synovial hyperplasia,while the excess of"yin forming elements"is the manifestation of synovial hyperplasia.Based on this,it put forward that"assisting yang qi as the priority,and according to the bias of pathogenic factors of yin,supplementing the method of reducing yin forming elements"is an important principle for treating this disease,which could provide new ideas for the treatment of the disease.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective To understand the changing composition and antibiotic resistance of bacterial species in the clinical isolates from outpatient and emergency department(hereinafter referred to as outpatients)and inpatient children over time in various hospitals,and to provide laboratory evidence for rational antibiotic use.Methods The data on clinically isolated pathogenic bacteria and antimicrobial susceptibility of isolates from outpatients and inpatient children in the CHINET program from 2015 to 2021 were collected and analyzed.Results A total of 278 471 isolates were isolated from pediatric patients in the CHINET program from 2015 to 2021.About 17.1％of the strains were isolated from outpatients,primarily group A β-hemolytic Streptococcus,Escherichia coli,and Staphylococcus aureus.Most of the strains(82.9％)were isolated from inpatients,mainly SS.aureus,E.coli,and H.influenzae.The prevalence of methicillin-resistant S.aureus(MRSA)in outpatients(24.5％)was lower than that in inpatient children(31.5％).The MRSA isolates from outpatients showed lower resistance rates to the antibiotics tested than the strains isolated from inpatient children.The prevalence of vancomycin-resistant Enterococcus faecalis or E.faecium and penicillin-resistant S.pneumoniae was low in either outpatients or inpatient children.S.pneumoniae,β-hemolytic Streptococcus and S.viridans showed high resistance rates to erythromycin.The prevalence of erythromycin-resistant group A β-hemolytic Streptococcus was higher in outpatients than that in inpatient children.The prevalence of β-lactamase-producing H.influenzae showed an overall upward trend in children,but lower in outpatients(45.1％)than in inpatient children(59.4％).The prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKpn),carbapenem-resistant Pseudomonas aeruginosa(CRPae)and carbapenem-resistant Acinetobacter baumannii(CRAba)was 14％,11.7％,47.8％in outpatients,but 24.2％,20.6％,and 52.8％in inpatient children,respectively.The prevalence of multidrug-resistant E.coli,K.pneumoniae,Proteus mirabilis,P.aeruginosa and A.baumannii strains was lower in outpatients than in inpatient children.The prevalence of fluoroquinolone-resistant E.coli,ESBLs-producing K.pneumoniae,ESBLs-producing P.mirabilis,carbapenem-resistant E.coli(CREco),CRKpn,and CRPae was lower in children in outpatients than in inpatient children,but the prevalence of CRAba in 2021 was higher than in inpatient children.Conclusions The distribution of clinical isolates from children is different between outpatients and inpatients.The prevalence of MRSA,ESBL,and CRO was higher in inpatient children than in outpatients.Antibiotics should be used rationally in clinical practice based on etiological diagnosis and antimicrobial susceptibility test results.Ongoing antimicrobial resistance surveillance and prevention and control of hospital infections are crucial to curbing bacterial resistance.

Objective To investigate the changing antibiotic resistance profiles of E.coli isolated from patients in the 52 hospitals participating in the CHINET program from 2015 to 2021.Methods Antimicrobial susceptibility was tested for clinical isolates of E.coli according to the unified protocol of CHINET program.WHONET 5.6 and SPSS 20.0 software were used for data analysis.Results Atotal of 289 760 nonduplicate clinical strains ofE.coli were isolated from 2015 to 2021,mainly from urine samples(44.7±3.2)％.The proportion of E.coli strains isolated from urine samples was higher in females than in males(59.0％vs 29.5％).The proportion of E.coli strains isolated from respiratory tract and cerebrospinal fluid samples was significantly higher in children than in adults(16.7％vs 7.8％,0.8％vs 0.1％,both P＜0.05).The isolates from internal medicine department accounted for the largest proportion(28.9±2.8)％with an increasing trend over years.Overall,the prevalence of ESBLs-producing E.coli and carbapenem resistant E.coli(CREco)was 55.9％and 1.8％,respectively during the 7-year period.The prevalence of ESBLs-producing E.coli was the highest in tertiary hospitals each year from 2015 to 2021 compared to secondary hospitals.The prevalence of CREco was higher in children's hospitals compared to secondary and tertiary hospitals each year from 2015 to 2021.The prevalence of ESBLs-producing E.coli in tertiary hospitals and children's hospitals and the prevalence of CREco in children's hospitals showed a decreasing trend over the 7-year period.The prevalence of CREco in secondary and tertiary hospitals increased slowly.Antibiotic resistance rates changed slowly from 2015 to 2021.Carbapenem drugs(imipenem,meropenem)were the most active drugs amongβ-lactams against E.coli(resistance rate≤2.1％).The resistance rates of E.coli to β-lactam/β-lactam inhibitor combinations(piperacillin-tazobactam,cefoperazone-sulbactam),aminoglycosides(amikacin),nitrofurantoin and fosfomycin(for urinary isolates only)were all less than 10％.The resistance rate of E.coli strains to antibiotics varied with the level of hospitals and the departments where the strains were isolated,especially for cefazolin and ciprofloxacin,to which the resistance rate of E.coli strains from children in non-ICU departments was significantly lower than that of the strains isolated from other departments(P＜0.05).The E.coli isolates from ICU showed higher resistance rate to most antimicrobial agents tested(excluding tigecycline)than the strains isolated from other departments.The E.coli strains isolated from tertiary hospitals showed higher resistance rates to the antimicrobial agents tested(excluding tigecycline,polymyxin B,cefepime and carbapenems)than the strains from secondary hospitals and children's hospitals.Conclusions E.coli is an important pathogen causing clinical infection.More than half of the clinical isolates produced ESBL.The prevalence of CREco is increasing in secondary and tertiary hospitals over the 7-year period even though the overall prevalence is still low.This is an issue of concern.

Objective To investigate the changes of diameter of the main trunk of the extracranial segment of the facial nerve,local hemodynamics and facial muscle thickness in patients with peripheral facial palsy(PFP)at acute stage by high frequency ultrasound(HFUS).Methods Sixty patients with acute PFP were enrolled as the PFP group,and 30 healthy people were selected as the control group.The following facial param-eters were scanned and recorded using HFUS:diameter of the main trunk of the extracranial segment of the facial nerve(FN-d),facial artery diameter(FA-d),facial artery systolic velocity(FA-Vs),facial artery diastolic velocity(FA-Vd),facial artery resistance index(FA-RI),frontalis muscle thickness(FM-t),depressor anguli oris muscle thickness(DAO-t),and depressor labii inferioris muscle thickness(DLI-t).The feasibility of facial param-eters in the diagnosis of acute-phase peripheral facial palsy(PFP)was evaluated,with intra-group and inter-group variability comparisons performed.Pearson correlation analysis was conducted to evaluate the relationship between acute-phase facial parameters and House-Brackmann(H-B)scale scores in PFP patients.Results(1)The H-B score of the PFP group was significantly lower than that of the control group,and the difference was statistically significant(P＜0.05).(2)The reliability of repeated measurements of facial parameters by different testers is good(ICC＞0.75).(3)The FN-d and FA-RI values on the affected side of PFP group were higher than those on the healthy side and control group,and FA-Vs,FA-Vd,DAO-t and DLI-t values were lower than those on the healthy side and control group,with statistical significance(P＜0.05).(4)The FN-d and FA-RI values of the affected side in the PFP group were negatively correlated with the H-S scale score(r=-0.847,P＜0.05;r=-0.863,P＜0.05);FA Vs,FA Vd,DAO-t,DLI-t are positively correlated with H-S scale scores(r=0.808,P＜0.05;r=0.757,P＜0.05;r=0.836,P＜0.05;r=0.694,P＜0.05).Conclusions HFUS can effectively detect the characteristic changes such as the increase in the diameter of the extrcranial facial nerve trunk,local microcircula-tion disturbance and facial muscle atrophy in patients with PFP in the acute stage,and the increase and decrease of FN-d,FA Vs,FA Vd,FA-RI,DAO-t,and DLI-t can accurately reflect the improvement or progress of PFP,which has important guiding significance for clinical disease monitoring and efficacy evaluation.

Objective:To explore the incidence, clinical features, genetic variant characteristics and prognosis of Glutaric acidemia type I (GA1) among neonates from Henan Province.Methods:A total of 814 625 neonates undergoing screening for inherited metabolic diseases by tandem mass spectrometry (MS/MS) at the Third Affiliated Hospital of Zhengzhou University from January 2016 to December 2022 were selected as the study subjects. A retrospective method was adopted to collect the clinical data of the patients. Whole exome sequencing was carried out to detect GCDH gene variants in individuals with positive results by GA1 newborn screening, and Sanger sequencing was used to verify the candidate variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of candidate variants was rated. This study was approved by the Medical Ethics Committee of the Hospital (Approval Number: 2019 Medical Ethics Review No. 67). Results:Eight cases of GA1 were diagnosed among the 814 625 neonates. Blood glutaryl carnitine (C5DC) and urine glutaric acid (GA) levels of the 8 children were higher than the normal reference values. In total 12 variants were detected, all of which were missense variants. c. 1064G>A (p.Arg355His) was the most common one, accounting for 21.4% (3/14). Three GCDH gene variants, including 1297G>C (p.Ala433Pro), c. 467G>A (p.Gly156Asp) and c. 1125T>G (p.Cys375Trp), were previously unreported. REVEL software analysis predicted that all of the three variants were harmful. 3D protein structure modeling indicated that the three variants may cause amino acid residue alterations, and c. 1297G>C (p.Ala433Pro) and c. 1125T>G (p.Cys375Trp) may result in increase in hydrogen bonds and may affect the function of GCDH protein. By December 2023, one of the eight children had deceased, and another child had severe clinical symptoms with poor prognosis. Six children had a good prognosis, of which two had mild motor development delay and four had normal development without clinical symptoms. Conclusion:The incidence of GA1 in newborns screened by MS/MS in Henan Province is 1/101 828, and the carrier rate of pathogenic GCDH variants is 1/160. The c. 1064G>A (p.Arg355His) may be the hotspot variant of the GCDH gene among children with GA1 in Henan. Discovery of the three novel variants has enriched the mutational spectrum of the GCDH gene and provide a basis for the early diagnosis, treatment, prognosis and genetic counseling of this disease.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.