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 investigate the serological characteristics and gene mutation mechanism of RhD variant blood donors in Zhengzhou.Methods From January 2023 to December 2023,1 619 RhD-negative blood donors sent to our labora-tory were selected for the study,and RhD negative confirmation test and RhCE phenotype detection were applied by tube method and microcolumn gel indirect antiglobulin test method.RHD gene amplification and Sanger sequencing were used to detect RhD variant sample genotypes.Results A total of 69 cases of RhD variants were detected in the RhD negative con-firmation test,with a proportion of 4.26%(69/1 619).The RhCE phenotypes were ccEe,Ccee,CcEe and CCee.There were 17 genotypes and 15 phenotypes of the D variant.The RHD?weak partial 15 allele was the most frequent(33 cases),with a frequency of 47.83%(33/69),and the main phenotype was the ccEe.This was followed by the RHD?DVI.3 allele in 20 ca-ses with a frequency of 28.99%(20/69)and the predominant phenotype was Ccee.The RHD?weak partial 15/RHD?01EL.01 heterozygote was found in 3 cases with a frequency of 4.35%(3/69),all with the CcEe phenotype.Other rare genotypes were present in 13 cases with a frequency of 18.84%(13/69).Antibody screening was positive in 3 cases with a frequency of 4.35%(3/69).Two cases of female blood donors,both with history of pregnancy and childbirth,were identified as anti-D;one case of male donor was anti-M.Conclusion The RHD?weak partial 15 genotype was the most common among the RhD variants in blood donors in Zhengzhou,followed by the RHD?DVI.3 genotype.It plays an important role in guarantee-ing the safety of blood supply and guiding precision transfusion.

Objective:To develop an HPLC method with gradient elution for the simultaneous determination of 4 bacteriostatic agents in triamcinolone acetonide econazole cream from the different manufacturers.Methods:The test was performed on a Waters Symmetry C18 column(250 mm ×4.6 mm,5 μm)under gradient elution of metha-nol(A)and methanol-0.02 mol·L-1 sodium dihydrogen phosphate solution(B).The flow rate of 1.0 mL·min-1 and the column temperature was 35 ℃.The detection wavelengths were 228 nm and 254 nm.Results:The good separation of the bacteriostatic agents peaks were achieved.The linear ranges of benzoic acid,methyl hydroxybenzoate,ethylparaben and propylparaben fell into 0.010 348-0.155 22 mg·mL-1,0.009 876-0.148145 mg·mL-1,0.010 106-0.151 588 mg·mL-1 and 0.010 259-0.153 882 mg·mL-1 respectively.The av-erage recoveries were 99.68％(RSD=2.35％),100.21％(RSD=1.78％),100.47％(RSD=1.59％)and 100.06％(RSD=1.65％)respectively.The LODs were 3 × 10-5 mg·mL-1,1.67 × 10-5 mg·mL-1,1.67 × 10-5 mg·mL-1 and 1.67 × 10-5 mg·mL-1 respectively.Conclusion:The established method is sensitive and accurate,and has the good separation.It provides the reliable basis for the quality control of triamcinolone ace-tonide econazole cream.

Objective To investigate the potential causal relationships between coronavirus disease-2019(COVID-19)susceptibility,hospitalization and severe case with late-onset myasthenia gravis(LOMG)based on Mendelian randomization(MR).Methods The public data from non-overlapping genome-wide association studies were screened,COVID-19 susceptibility,hospitalization and severe case served as the exposure data,and LOMG as the outcome data.The inverse-variance weighted(IVW)method was mainly adopted to evalu-ate the causal effect,which was supplemented by the methods such as MR-Egger method,weighted median method,weighted model and simple model.The sensitivity analysis was performed.Results The genetically predicted severe case of COVID-19 had the positively causal relationship with LOMG(OR=1.01,95％CI:1.00-1.03,P=0.046).The sensitivity analysis results revealed the study results were steady(P＞0.05).No heterogeneity or horizontal pleiotropy was found.Conclusion Severe case of COVID-19 may be associated with an increased risk of LOMG.

Objective:To serologically and genotypically analyze the pedigree of a case with a new allele c.175_176insGA of B el subtype and preliminarily explore the molecular mechanism of weak expression of glycosyltransferase B. Method:In the descriptive study,a 23-year-old male voluntary blood donor and his family members were selected for the study. The ABO and Le blood types of the proband and his family members was identified by the test tube method. The agglutination inhibition test was applied to detect the B and H antigens in saliva, and the Sanger sequencing and PacBio (Pacific Bioscience) third-generation haplotype sequencing were performed on the study subjects to identify genotypes. Finally, Expasy software were applied to amino acid translation of DNA sequences and prediction of protein length after gene alteration. ORF finder was applied to predict alternative start codons as well as open reading frames of mRNA, and protein expression mechanisms were analyzed.Results:The proband and her sister were B el subtype, her mother was AB el subtype, her father was normal O type, and all members of the family were Le(a+b+) phenotype. Sanger sequencing results showed that a new allele of c.175_176insGA was found in exon 4 of the proband, her mother, and her sister. Third-generation haplotype sequencing detected the haplotypes of the family members, which revealed that the proband was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA), the father was ABO*O.01.02/ABO*O.01.02, the mother was ABO*A1.02/ABO*BEL.NEW (c.175_176insGA), and the sister was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA). Analysis of the protein expression mechanism indicated that although the new allele of ABO*BEL.NEW was presumed to cause a frameshift mutation and result in a premature stop codon p.Asp59Glu*fs20 in exon 5, encoding an inactive glycosyltransferase, an alternative start codon could be utilized to initiate translation of B el subtype functional glycosyltransferase. Conclusion:Expression of the new allele of B el subtype is associated with the translation of B el subtype glycosyltransferase initiated by alternative start codons.

Objective To explore the comorbidity patterns of chronic diseases and their associated factors in the elderly population aged 60 and above in China. Methods Based on the data from the 2018 China Health and Retirement Longitudinal Study (CHARLS), the Apriori algorithm in R4.0.4 software was used to extract frequent itemsets and construct an association rule model between chronic diseases and their associated factors, targeting the information on 14 chronic diseases in people aged 60 and above in the database. Results Among the 13 206 participants, 3 691 cases (27.90%) suffered from at least one chronic disease, while 6 217 cases (47.08%) suffered from two or more chronic diseases. Analysis of association rules showed that the most common comorbidity patterns included heart disease and hypertension, dyslipidemia and hypertension, diabetes and hypertension, as well as "heart disease, arthritis→high blood pressure" "high blood pressure, stomach disease→arthritis" "dyslipidemia, arthritis→hypertension", with support degrees of 8.93%, 8.29%, 5.44%, 3.82%, 3.71% and 3.22%, and confidence degrees of 55.15%, 59.64%, 60.13%, 53.67%, 51.58% and 60.11% respectively. Irregular sleep patterns, urban living backgrounds, and high social participation were strongly associated with hypertension; while irregular sleep patterns, rural residency, and female were highly associated with arthritis. Conclusion Given the universality of hypertension in multiple chronic disease comorbidity patterns, the prevention and screening of hypertension and its complications in the elderly population should be strengthened. For the comorbidity patterns of chronic diseases coexisting with hypertension, special attention should be paid to the daily disease screening and management of the elderly who participate in social activities in cities. At the same time, for elderly women, poor sleep quality, and rural residents, the screening and attention to the comorbidity patterns of chronic diseases with arthritis need to be strengthened.

【Objective】 To carry out serological and molecular biological identification of B (A) subtype, and discuss the rational blood transfusion strategy. 【Methods】 Serological and direct sequencing methods were used to detect serotype and genotype of 7 cases of B (A) subtype, and cross matching was performed by saline medium and anti human globulin card to analyze the red blood cells(RBCs) transfusion strategy. 【Results】 The serology results of blood type of 7 samples were similar, with B(A)04/O01 in 3 cases, B(A)04/O02 in 2 cases and B(A)02/O01 in 2 cases. 7 cases of B (A) subtypes were matched with randomly selected blood donors of type O and B on the major side. 【Conclusion】 B(A) subtypes should be identified by genotyping techniques. Washed RBCs of type B and O can be used for B(A) blood type transfusion.

Objective:To investigate the characteristics of anxiety and depression of postgraduates in military medical university under public health emergencies.Methods:By means of self-rating anxiety scale (SAS) and self-rating depression scale (SDS), 944 postgraduate students of a military medical university were investigated to analyze their characteristics of anxiety and depression. SPSS 25.0 was used to statistically analyze the collected data.Results:①The positive rates of anxiety and depression were respectively 8.03% and 8.25%. ②The positive rate of anxiety of male postgraduates was significantly higher than that of female postgraduates ( P=0.035). ③The positive rate of anxiety of serviceman students was significantly higher than that of civilian students ( P=0.022). ④There was no statistical difference in the positive rates of anxiety and depression among postgraduates in different areas ( P > 0.05). ⑤The levels of anxiety and depression of left-behind children were significantly higher than those of unleft-behind children ( P=0.000, P=0.018). ⑥The levels of anxiety and depression of postgraduate students were lower than those of undergraduate students ( P=0.000, P=0.002). Conclusion:Under the outbreak of public health emergencies, anxiety and depression occur in the postgraduates. To strengthen the mental health maintenance of postgraduates in military medical university during the outbreak, it is necessary to pay more attention on the mental health of military students, males and who were left-behind children before.

【Objective】 To analyze and master the serological and genetic characteristics of the samples with CisAB subtype and their genetic background. 【Methods】 From January 2018 to January 2022, blood samples with discrepant ABO blood typing results, from Zhengzhou voluntary blood donors and hospital patients, were subjected to phenotypic classification using micro column gel card and tube method, as well as amplification of exons 6 and 7 in ABO gene using PCR. The pedigrees of individuals with the same CisAB subtype but different serological typing results in the same family were analyzed. 【Results】 11 The forward typing of 12 samples was AB type, and unexpected antibodies against weaker antigens were found in 11 serum samples, including 9 cases with strong antigen A, 2 cases with strong antigen B, and 1 case with consistent forward and reverse typing results. Gene sequencing confirmed that 11 cases were CisAB01 subtype and 1 case was CisAB05 subtype.Among them, 7 cases had the genotype of CisAB01/O and serological phenotype of A2B3; 2 cases had the genotype of CisAB01/B and phenotype of A2B; 2 cases had the genotype of CisAB01/A and serological phenotype of A1Bx and A1B3; 1 case had the phenotype of AxB. In the CisAB01 family, 1 case of CisAB01/O with A2B3 phenotype and 1 case of CisAB01/B with A2B phenotype were detected. In the CisAB05 family, 2 CisAB05/O01 and 1 CisAB05/O02 were detected. 【Conclusion】 The serological phenotypes of different individuals in the same CisAB01 family can be different when paired with different ABO alleles. It is advisable to accurately identify the CisAB subtype genes with molecular biological methods to ensure blood transfusion safety.

Objective To investigate the clinical value of 99Tcm-methoxyisobutylisonitrile (MIBI) double-phase imaging in differential diagnosis of bone lesions in tumor patients,which was indefinite in 99Tcm-methylene diphosphonate (MDP) bone imaging,and evaluate the correlation between 99Tcm-MIBI imaging and chemotherapy efficacy.Methods Fifty-two tumor patients (23 males,29 females,mean age 58 years) with bone isolated lesions(≤3) found by 99Tcm-MDP bone scintigraphy was enrolled from June 2014 to November 2015.Since the results of 99Tcm-MDP imaging were indefinite,99Tcm-MIBI double-phase (10 min and 30 min after injection of 99Tcm-MIBI) SPECT/CT imaging was then performed within 1 week.The final diagnosis was made according to results of more than two imaging modalities (CT,MRI,PET/CT) and/or follow-up (≥6 months).The diagnostic efficacy of 99Tcm-MIBI SPECT/CT imaging was calculated.The clinical dataof chemotherapy were also collected.Patients with bone metastasis were grouped as the treatment response.Retention index (RI) of metastatic lesions was calculated and compared between different groups using two-sample t test.The relationship between RI and chemotherapy efficacy was investigated by Spearman correlation analysis.Results A total of 12 benign lesions and 84 malignant lesions were eventually diagnosed.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and Youden index of 99Tcm-MIBI imaging were 96.43％(81/84),83.33％(10/12),97.59％(81/83),76.92％(10/13),and 0.80,respectively.The RI was statistically different between complete remission (CR)+partial remission (PR) group and stable disease (SD) + progressive disease (PD) group:-0.142± 0.036 vs-0.384± 0.067 (t =2.367,P＜0.05).The RI of 99Tcm-MIBI in bone metastases was positively correlated with the chemotherapy efficacy (rs =0.78,P＜0.01).Conclusions 99Tcm-MIBI imaging is helpful in differential diagnosis of bone lesions with indefinite diagnosis by 99Tcm-MDP bone imaging in tumor patients.99Tcm-MIBI RI of bone metastasis may predict the therapeutic response of chemotherapy efficacy.