BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

Objective:To investigate the incidence of radiation pneumonitis (RP) induced by 125I seed implantation for the treatment of malignant lung tumors and analyze related dosimetric parameters. Methods:A retrospective analysis was conducted on 31 cases of malignant lung tumors treated with 125I seed implantation from January 2017 to December 2022 at Hebei Provincial Tumor Radioactive Seeds Implantation Diagnosis and Treatment Center. These cases consisted of eight patients with squamous cell carcinoma, 10 patients with adenocarcinoma, and 13 patients with metastatic cancer in other sites. At 1-6 months after treatment, these patients received postoperative chest CT scans, with the efficacy evaluated based on the Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST 1.1), including the objective response rate (ORR) and the disease control rate (DCR). The efficacy of RP was evaluated using the Radiation Therapy Oncology Group (RTOG) criteria. Postoperative dosimetric parameters, including D90 (minimum peripheral dose received by 90% of the target volume), V8 (percentage of lung volume receiving 8 Gy), V32 (percentage of lung volume receiving 32 Gy), and Dmean (mean radiation dose) of the affected lung, were statistically analyzed. The relationships of the RP occurrence with postoperative D90, V8, V32, and Dmean were analyzed by comparison with relevant external radiotherapy data, to identify the parameters that are correlated closely with RP occurrence. Results:All the patients underwent successful surgeries. The postoperative efficacy evaluation after six months showed complete response (CR) in 11 cases, partial response (PR) in 11 cases, stable disease (SD) in eight cases, and progressive disease (PD) in one case, with an overall response rate (ORR) of 71.0%, and a disease control rate (DCR) of 96.8%. Three patients suffered RP, with an incidence rate of 9.7%. Postoperative V8, V32, and Dmean could not serve as predictive indicators for RP. Follow-up observation revealed that three RP cases (3/5) exhibited postoperative D90 exceeding 170 Gy and no RP cases (0/26) showed postoperative D90 below 170 Gy. Conclusions:In the treatment of malignant lung tumors with 125I seed implantation, there is a certain correlation between RP and postoperative D90, while there is no correlation between it and V8, V32, and Dmean.

Objective:To analyze the effect of montelukast combined with budesonide in the treatment of children with intermittent asthma, and the impact on airway remodeling and T helper type 1 (Th1)/T helper type 2 (Th1/Th2) related cytokines.Methods:A prospective study was conducted among 120 children with intermittent asthma admitted to Huanghua Municipal People′s Hospital from December 2021 to February 2023. The children were randomly divided into the control group (60 children treated with budesonide atomizationinhalation) and the observation group (60 children treated with montelukast on the basis of the treatment of control group). Clinical efficacy, airway remodeling indicators [total area of airway (Ao), outer diameter of airway (D) and wall area to total airway cross-sectional area (WA%)], pulmonary function [peak expiratory flow (PEF), forced expiratory volume in 1 second (FEV 1)/forced vital capacity (FVC) and the maximum expiratory flow at 25% of vital capacity (MEF25%)], Th1/Th2 related cytokines, inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), interleukin-6 (IL-6) and interferon-gamma (IFN-γ)], recurrence, and the incidence of adverse reactions were compared between the two groups. Results:The total effective rate in the observation group was higher than that in the control group: 90.00% (54/60) vs. 75.00% (45/60) ( P<0.05). After treatment, Ao, D and WA% in the observation group were lower than those in the control group: (17.58 ± 1.89) mm 2 vs. (19.22 ± 1.94) mm 2, (4.25 ± 0.48) mm vs. (4.48 ± 0.49) mm, (63.75 ± 6.49)% vs. (69.22 ± 7.14)% ( P<0.05). PEF, FEV 1/FVC and MEF25% in the observation group were higher than those in the control group: (3.13 ± 0.34) L/s vs. (2.86 ± 0.35) L/s, (87.45 ± 8.86) % vs. (83.59 ± 8.42) %, (87.63 ± 8.86)% vs. (82.15 ± 8.43)% ( P<0.05). The levels of Th1 and Th1/Th2 in the observation group were higher than those in the control group: (14.13 ± 1.46) % vs. (10.27 ± 1.25) %, 3.46 ± 0.39 vs. 1.88 ± 0.25, and the level of Th2 was lower than that in the control group: (3.96 ± 0.45)% vs. (5.48 ± 0.56)% ( P<0.05). After treatment, the levels of TNF-α and IFN-γ in the observation group were higher than those in the control group: (76.15 ± 7.78) ng/L vs. (66.38 ± 6.47) ng/L, (7.15 ± 0.74) ng/L vs. (6.14 ± 0.66) ng/L. The levels of IL-4 and IL-6 were lower than those in the control group: (77.85 ± 7.96) ng/L vs. (86.42 ± 8.74) ng/L, (37.25 ± 3.89) mg/L vs. (44.23 ± 4.57) mg/L ( P<0.05). The recurrence rate in the observation group was lower than that in the control group: 3.33% (2/60) vs. 15.00% (9/60) ( P<0.05). The incidence rates of adverse reactions in the two groups were without statistically significant difference between the groups ( P>0.05). Conclusions:Montelukast combined with budesonide can reduce airway remodeling in children with intermittent asthma, improve their pulmonary function, Th1/Th2 related cytokines and inflammatory response indicators, and reduce recurrence rate, with good safety.

Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*

Objective To analyze the epidemiological characteristics of mobile colistin resistance(mcr)genes in global Klebsiella pneumoniae(K.pneumoniae).Methods The genomes of K.pneumoniae were downloaded from the NCBI genome database by the As-pera software.After quality filtering using the CheckM v1.1.3 and Quest 5.0.2 software,the genomes were annotated using the Prokka v1.13.All the mcr gene sequences were downloaded from the NCBI website and a database was built using the makeblastdb command.Then,a self-made Perl script program was used to extract the nucleotide sequences of all genes from the annotation file as Query,and the local BLASTN analysis was performed to obtain mcr-positive strains.The gene sequence files and Profiles files of 7 housekeeping genes of K.pneumoniae were downloaded from the PubMLST website as the database,and the nucleotide sequences of the genes were extracted as Query using a self-made Perl script program.The local BLASTN analysis was implemented to determine the sequence type(ST)of each genome.Meta information of each strain,including isolation source,sample type,country,and date,were extracted in batch from the GenBank file of the downloaded K.pneumoniae genomes using a self-written Perl program to analyze the distribution characteristics of mcr-positive strains.The distribution differences of the mcr between different ST types were compared by the Chi-square test.Results Among the 11 429 global K.pneumoniae genomes included in this study,229 mcr genes were detected from 207 strains.Six variants of mcr were identified,mainly mcr-1(87/229,38.0％),mcr-8(59/229,25.8％),and mcr-9(59/229,25.8％).76 STs were identified from 207 strains,with ST15(21/207,10.1％),ST43(17/207,8.2％),ST11(16/207,7.7％),and ST 147(16/207,7.7％)being the predominant.Among 87 mcr-1 positive strains,31 STs were found,with ST43(17/87,19.5％)and ST15(10/87,11.5％)being the main ones.Among 59 mcr-8 positive strains,17 STs were identified,with ST43(17/59,28.8％)and ST11(9/59,15.3％)being the predominant.Among 59 mcr-9 positive strains,27 STs were detected,with ST147(11/59,18.6％)and ST274(11/59,18.6％)being the main ones.There were statistical differences in the variants of mcr genes carried by different ST types.The mcr-positive K.pneumoniae came from 28 countries across five continents worldwide,led by China(68/207,32.9％)and Thailand(45/207,21.7％),which were mainly from the human body(100/207,48.3％)and had a concentrated outbreak time from 2015 to 2018.Conclusion Among the global K.pneumoniae,the prevalence of mcr is mainly domina-ted by mcr-1,mcr-8,and mcr-9.The dominant clones of mcr-1 are mainly ST15 and ST43,while those of mcr-8 are mainly ST11 and ST43.The popularity of mcr-9 is mainly based on ST147 and ST274.Strengthening the monitoring of such bacteria may play an impor-tant role in preventing and controlling nosocomial infections.

Objective To analyze the prevalence and molecular characteristics of global Salmonella Typhimurium.Method The ge-nome and the corresponding serovars as well as meta-information of global Salmonella were downloaded in batch from the National Cen-ter for Biotechnology Information(NCBI)database using the online softwares,ResFinder,PlasmidFinder,Mobile Element Finder and MLST,to analyze the distribution of antibiotic resistant genes(ARGs),plasmids,mobile genetic elements(MGEs)and the sequence types(STs)among S.Typhimurium strains.Results A total of 101 ARGs were detected in 323 strains of S.Typhimurium,among which the most common was aac(6')-Iaa(322/323,99.69％),followed by sul2(155/323,47.99％),aph(3")-Ib(128/323,39.63％),aph(6)-Id(127/323,39.32％),tet(B)(109/323,33.75％),and blaTEM-1B(106/323,32.82％).Totally,36 plasmids were identified,among which IncFⅡ(S)(110/323,34.06％)and IncFⅠB(S)(108/323,33.44％)were the most common.Moreo-ver,376 MGEs were found,including 367 insertion sequences and nine transposons,among which MITEEc1(322/323,99.69％)and ISSen1(308/323,95.36％)were the most popular.Furthermore,323 strains of S.Typhimurium were assigned into 32 different STs,a-mong which,ST19(157/323,48.61％)and ST34(105/323,32.51％)were the most common accounting for over 82％.Among 115 strains of S.Typhimurium from human,18 STs were identified including ST19(52/115,45.22％)and ST34(39/115,33.91％)which were the most common.Conclusion S.Typhimurium carried multiple types of ARGs,in addition to the wide distribution of a large number of insertion sequences and plasmids,whch provide favorable conditions for the spread of drug resistance.Therefore,the meas-ures of prevention and control against the infection from this bacterium should be strengthened.

Objective To analyze the application of serological test results in the diagnosis and treatment of anti-M-in-duced hemolytic disease of the fetus and newborn(HDFN),and to explore HDFN prevention strategies.Methods The se-rological test results of 12 cases of HDFN caused by anti-M diagnosed in our laboratory from January 2017 to December 2023 were retrospectively analyzed,including blood group identification of mothers and children,serum total bilirubin/hemoglo-bin/antibody titer test,and three hemolysis tests in newborns.Clinical data of the children and mothers were collected,in-cluding pregnancy history,blood transfusion history,prenatal antibody testing,history of intrauterine blood transfusion and gestational week of delivery,and the prognosis of the children was followed up.Results All 12 cases of fetal neonatal he-molytic disease due to anti-M were RhD+MN phenotype newborn born to RhD+NN mother,with maternal-fetal incompati-blility in MN blood groups.In the ABO blood group system,ABO incompatibility between mother and child accounted for 41.7%(5/12).None of the mothers had a history of blood transfusion,and the median titer of the test at 4℃was 32,and the median titer at 37℃was 4.The mothers of 3 cases had a history of multiple intrauterine blood transfusions,with an inci-dence of 25%(3/12).One case had an abnormal first pregnancy,with an incidence of 8.3%(1/12),and seven cases had an abnormal pregnancy with a miscarriage,with an incidence of abnormal pregnancy and birth history of 58.3%(7/12).There were 6 cases of premature labor,with an incidence of 50%(6/12).The mothers in three cases underwent regular ob-stetric examination and the specificity of the antibodies was determined,accounting for 25%(3/12).Twelve children had free antibodies with a median titer of 6 at 4℃and 2 at 37℃.Two children had anti-M antibodies that were not reactive at 37℃,with a negative rate of 16.7%(2/12).The positive rate of DAT and elution test was respectively 8.3%(1/12)and 16.7%(2/12)in the children.The median minimum hemoglobin value was 75 g/L,and all 12 children received blood transfusions.The median peak total bilirubin value was 157.5 μmol/L,and none of them reached the threshold for blood ex-change.The rate of delayed anemia was16.7%(2/12),the postnatal mortality rate was8.3%(1/12),and 11 children was free of growth and neurodevelopmental delay in prognosis.Conclusion Anti-M can cause severe HDFN,which can also oc-cur in primigravida.The intensity of antibody titer does not correlate with the severity of the disease,and it is prone to cause delayed anemia,which should be monitored regularly according to the serological characteristics of anti-M and clinical symp-toms,and should be treated timely.

Objective To analyze the risk factors for iatrogenic pseudoaneurysm(PSA)occurring after cardiovascular interventional procedures.Methods The clinical data of 48 patients,who developed PSA after receiving cardiovascular interventional procedure at the Yantai Yuhuangding Hospital of China between January 2018 and December 2022,were retrospectively analyzed.The control group included 192 patients who had no PSA.At a case-control ratio of 1∶4,the PSA patients and non-PSA patients were paired,and the paired indicators included age,and puncture site.Univariate and multivariate logistic regression analyses were used to analyze the patients'basic data,hematological examination,and situation of the interventional procedure,and the independent risk factors were screened out.Results Multivariate logistic regression analysis showed that the high body mass index(BMI,OR=1.324,95％CI=1.097-1.598,P=0.003),smoking history(OR=4.477,95％CI=1.599-12.536,P=0.004),use of antiplatelet agents(OR=4.861,95％CI=1.018-23.214,P=0.047),combination use of antiplatelet and anticoagulant(OR=26.994,95％CI=2.353-309.686,P=0.008),the operator of the interventional procedure being an attending physician(OR=5.817,95％CI=1.139-29.717,P=0.034),low haemoglobin level(OR=0.946,95％CI=0.922-0.971,P＜0.01),elevated D-dimer level(OR=2.407,95％CI=1.367-4.239,P=0.002),long-time interventional operation(OR=1.019,95％CI=1.005-1.033,P=0.009),and sheath size＞6 F(OR=4.368,95％CI=1.196-15.947,P=0.026)were the independent risk factors for PSA occurring after cardiovascular interventional surgery.Conclusion High BMI,smoking history,use of antiplatelet agents,combination use of antiplatelet and anticoagulant,the operator of the interventional procedure being an attending physician,low haemoglobin level,elevated D-dimer level,long-time interventional operation,and sheath size＞6 F are the independent risk factors for PSA occurring after cardiovascular interventional procedure,which can provide a basis for the early prevention of PSA.(J Intervent Radiol,2024,33:646-650)

Objective To analyze the prevalent characteristics,sequence type(ST),and blaOXA gene distribution of global Acineto-bacter pittii,and provide reference for prevention and clinical medication against this infection.Methods All the sequences of A.pittii genomes were downloaded in batch from NCBI by using Aspera software with deadline of November 30,2023,and perl script was used to extract the meta information of strains in batch from the downloaded GBK file.A self-made Perl program was utilized to extract the nucleotide coding sequence of the gene from each genome sequence file of A.pittii as the analysis database.BLASTN comparative analy-sis was implemented by using the allelic sequences of 7 housekeeping genes of the genus Acinetobacter Sp downloaded from the website as a query database to determine the ST of each genome.The nucleotide sequences of the blaOXA genes were downloaded from the NCBI pathogen resistance gene database and the self-written AMRG software was utilized to analyze the distribution of blaOXA gene among A.pittii.Results A total of 305 A.pittii genomes were obtained,which were mainly isolated from 1990 to 2020.The isolation rates showed an increasing trend of year by year with the highest isolation rate in 2015.The separation rates of the United States,China and Germany ranked in the top three positions,with the separation rates of 29.5％(90 strains),15.7％(48 strains),and 13.4％(41 strains),respectively.A total of 180(59.0％)specimens were sourced from humans,of which 42(23.3％)were from sputum,27(15.0％)from blood,and 15(8.3％)from skin and soft tissues.The 305 strains of A.pittii were assigned into 79 STs,among which ST93(14.4％),ST64(12.1％)and ST63(11.8％)accounted for the highest proportion.In the United States,China and Germany,the infections were mainly affected by ST64,ST63 and ST93,respectively.The 305 A.pitti carried the blaOXA gene except 6 strains,while the remained 299 strains carried 31 blaOXA variants,of which,252 blaOXA genes were found to own the carbapenemase hydrolysis activity and 146 strains carried blaOXA-500 gene.Conclusion There should be regional differences in the global distribution of A.pittii,and the high prevalence of the carbapenemase gene blaOXA suggested a potential trend of multi-drug resistance,which is worthy to be monitored.

Objective To detect and analyze the drug resistance characteristics,phylogenetic typing,and virulence gene distribution of Escherichia coli(E.coli)in blood culture.Methods The strains of E.coli isolated from consecutive non-repetitive blood cultures in our hospital from January 1,2019 to December 13,2020 were collected.The sensitivity of E.coli to 17 antibiotics was determined u-sing the micro-broth method.The bacterial genomic DNA was extracted using the boiling method,and then the arpA,chuA,yjaA,TspE4C2,ArpAgpE and trpAgpC genes were detected by PCR to determine the bacterial phylogroup.The virulence genes,including iutA,fimH,fyuA,kpsMT Ⅱ,cnf1,cvac,hlyA,traT,kpsMT Ⅲ,and PAI,were detected using the multiplex PCR.The differences in drug resistance and virulence gene distribution among different phylogroups were analyzed by the Chi-square test.Results 270 strains of E.coli in blood culture showed high resistance rates to ceftriaxone,compound sulfamethoxazole,ampicillin,ampicillin sulbactam,cefazolin,and ciprofloxacin,all exceeding 50.0％.They had good susceptibility to imipenem,ertapenem,amikacin,and piperacillin tazobactam,with resistance rates all below 5.0％.The most common phylogroups were types B2 and D,accounting for 38.0％and 16.2％,respectively,while type E and hidden branch type I were relatively rare,accounting for less than 1.0％.The virulence gene analysis revealed that the distribution rates of fimH and fyuA genes were the highest,both above 99.0％.The distribution rates of kpsMT Ⅲ,hlyA,and cvaC genes were relatively low,all below 20.0％.The Chi-square test showed that the distribution rates of viru-lence genes such as iutA,fimH,fyuA,kpsMT Ⅱ,cnf1,and PAI in the B2 group were significantly higher than those in the non-B2 group(P<0.05).The distribution rates of iutA,fyuA,kpsMT Ⅱ,cnf1,and PAI genes in the B2 group were significantly higher than those in the D group(P<0.05).Conclusion When treating bloodstream infections caused by E.coli,caution should be exercised in the use of drugs such as ceftriaxone,compound sulfamethoxazole,ampicillin,ampicillin sulbactam,cefazolin,and ciprofloxacin.When bloodstream infections are caused by phylogroup B2 E.coli,middle-stream urine culture should be performed simultaneously to confirm the source of infection and monitor the success rate of treatment.