Objective: To design and establish a new method for flow cytometry-based detection of commonly observed highly expressed antigens on red blood cells, and to further evaluate the differences and distribution characteristics of antigen expression levels between ABO blood type homozygotes and heterozygotes in healthy individuals. Methods: Residual blood samples after donor blood type identification by Shanghai Blood Center in April 2024 were collected. Among them, samples of 19 homozygous and 19 heterozygous individuals of type A and type B were selected. Then the expression level of ABO antigen on red blood cells were detected using the new method established in this study and the traditional aldehyde fixed red blood cell method. Both methods were tested independently three times and the results were compared. Results: The mean values of the three detection results of the new method was (×10 /RBC): AA homozygous 3.3±0.5, AO heterozygous 2.8±0.3, BB homozygous 3.6±0.3, BO heterozygous 3.1±2.8. The mean values of the three detection results of the aldehyde fixation method were AA homozygous 5.9±0.9, AO heterozygous 5.0±1.4, BB homozygous 3.8±0.6, and BO heterozygous 3.3±0.4. The average antigen distribution of each genotype followed a normal distribution. Comparing the average antigen expression levels of homozygotes and heterozygotes, both methods showed that A/B homozygotes had higher antigen levels than heterozygotes, with AA being 1.17 to 1.18 times that of AO and BB being 1.15 to 1.16 times that of BO. Comparing the inter batch differences in the three test results of two methods, the new method showed no significant difference in the three test results for four genotypes (P>0.05). The aldehyde fixation method showed significant differences in the test results for all three genotypes (P<0.01) except for BB homozygotes (P>0.05). The reliability and reproducibility of the new method were better than those of the traditional aldehyde fixation method. Conclusion: The antigen expression level of ABO homozygotes is higher than that of heterozygotes, and the difference in antigen level between type A homozygotes and heterozygotes is slightly higher than that of type B. The new method is superior to traditional aldolization fixation methods.

Humans ; Female ; Blood Platelets ; Ovarian Neoplasms/diagnosis*

【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.

Small cell lung cancer (SCLC) is the most malignant lung cancer with the highest mortality. At present, the first-line standard treatment is still based on Etoposide and Platinum chemotherapy. However, for SCLC that progresses after first-line therapy, the treatment options are still very limited. Since the molecular mechanism of first-line drug resistance of SCLC is still unclear, and the precision medicine strategy after first-line drug resistance is still in the pre-clinical stage. The proportion of secondary biopsy and genetic testing is very low after the progress of first-line treatment of SCLC. In this study, we report a case of a middle-aged woman who was first diagnosed with SCLC. Adenocarcinoma with sensitive gene mutations and repeated changes of small cell carcinoma were detected by multiple biopsies during the course of the disease, suggesting that the patient may be a special subtype of SCLC - mixed SCLC (M-SCLC). In this case, the patient has been treated with radiotherapy and chemotherapy, immunotherapy and targeted therapy successively, and the survival time has reached 2 years and 8 months. Through the case report and literature review retrospectively, this study aimed to explore the part patients may start to present hybrid histopathologic types or tissue type change after treatment of SCLC. Biopsy pathologic histology and genetic testing is necessary after disease progression to look for potential therapeutic targets, so as to give precise treatment based on molecular markers detection results and provide the patient with the benefit of survival for as long as possible.
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Adenocarcinoma of Lung ; Etoposide ; Female ; Humans ; Lung Neoplasms/genetics* ; Middle Aged ; Retrospective Studies ; Small Cell Lung Carcinoma/genetics*

OBJECTIVE：To in vestigate the effects of quercetin （Que）on the expressio n of angiotensin Ⅱ（AngⅡ）-induced myocardial contractile proteins of primary rats through angiotensin-converting enzyme 2-angiotensin-（1-7）-Mas （ACE2-Ang- （1-7）-Mas）axis. METHODS ：Cardiac tissue of rats aged 1-2 d were collected ，and primary cardiomyocytes were isolated and cultured. The gene silencing model of cardiomyocytes ACE2 was constructed. Experiments were divided into 12 groups. Among them，AngⅡ group，AngⅡ+ small interference RNA （siRNA）group，and Ang Ⅱ+ A 779 group were the model groups ；AngⅡ+ losartan group was positive control group ；AngⅡ+Que40 group，AngⅡ+Que80 group，AngⅡ+siRNA+Que40 group，AngⅡ+ siRNA+Que80 group，AngⅡ+A779+Que40 group and Ang Ⅱ+A779+Que80 group were the experimental groups ；blank group and siRNA group were set up. Ang Ⅱ concentration was 1×10－6 mol/L；siRNA final concentration was 50 nmol/L；Que concentration was 40 and 80 μmol/L；A779（Mas receptor inhibitor ）concentration was 1 μmol/L；losartan concentration was 1×10－4 mol/L. mRNA and protein expression of ACE 2，Ang-（1-7） and Mas in primary cardiomyocytes were detected ；the expressions of myocardial contractile proteins were also determined ，such as Na +/Ca2+ exchange channel （NCX），calcium pump （SERCA2a）， phosphoprotein （PLB）. RESULTS ：Compared with Ang Ⅱ group，mRNA expression of Mas was increased significantly in Ang Ⅱ + Que 80 group （P＜0.05）；mRNA expression of ACE2 and Mas were increased significantly in Ang Ⅱ + CZ0210-01） losartan group （P＜0.05）. Compared with Ang Ⅱ group，the 851136165@qq.com protein expression of ACE 2 and Ang- （1-7） were increased significantly in Ang Ⅱ+ Que 40 group（P＜0.05）；compared with Ang Ⅱ + siRNA group ，the protein expression of Ang-（1-7）were increased significantly in Ang Ⅱ+ siRNA+Que 40 group（P＜0.05）；compared with Ang Ⅱ+A779 group，the protein expression of Ang- （1-7）were increased significantly in Ang Ⅱ+A779+ Que 40 group（P＜0.05）. Compared with Ang Ⅱ group，the protein expression of NCX was decreased in Ang Ⅱ+Que40 group（P＜0.05），protein expression of NCX was reduced in Ang Ⅱ+ losartan group （P＜0.05）；compared with Ang Ⅱ+A779 group，the protein expression of NCX was decreased in Ang Ⅱ+A779+ Que80 group （P＜0.05）. CONCLUSIONS ：Que improves the expression of Ang Ⅱ -induced ACE 2-Ang-（1-7）-Mas axis in cardiomyocyte model to some extent ，so as to regulate myocardial contractile protein.

Objective:To explore the effects of father-feeding attitudes on feeding status in preterm infants at discharge, and the role of father support breastfeeding self-efficacy, in order to provide a certain reference and basis for the intervention to improve the breastfeeding rate of premature infants.Methods:This was a cross-sectional study. A total of 402 premature infants' fathers were selected from 3 ClassⅢ Grade A hospitals from Zhejiang Province by convenience sampling method from July to November 2020. The basic data questionnaire, Iowa Infant Feeding Attitude Scale and Father Support Breastfeeding Self-Efficacy Scale-Short Form were used in the investigation. A total of 402 questionnaires were distributed, and 376 valid questionnaires were recovered. The recovery rate of effective questionnaires was 93.5%.Results:The score of father-feeding attitude of the 376 premature infants' fathers was (49.80±9.26) , and the score of support breastfeeding self-efficacy was (43.23±8.52) . Father's self-efficacy of breastfeeding support played a mediating role between feeding attitude and feeding status of premature infants at discharge, and the mediating effect accounted for 33.3% of the total effect.Conclusions:The father's feeding attitude can affect the feeding status of premature infants at discharge by influencing their self-efficacy of supporting breastfeeding. Medical staff should help premature fathers pay attention to breastfeeding, adopt practical and diverse health education methods to help premature fathers gradually master specific support behaviors and enhance their self-efficacy in supporting breastfeeding, and ultimately improve the rate of breastfeeding in premature infants.

Objective To investigate the correlation between carotid intima-media thickness (CIMT) and left atrial and left ventricular enlargement in patients with acute cerebral infarction. Methods A total of 224 patients with acute cerebral infarction were included. Based on the thickness of CIMT, it was divided into three groups which were normal CIMT group, thickening CIMT group, and carotid plaque (CP) group, with 57, 97, and 70 patients included respectively. Clinical data were collected, and carotid artery color Doppler ultrasound, cardiac color Doppler ultrasound and other examinations were determined to carry out relevant statistical analysis.Results The left anterior-posterior diameter (LAD) , left atrial diameter index (LADI) , left ventricular end-diastolic septal thickness (IVSD) , and left ventricular mass index (LVMI) in the CP group were all higher than those in the normal CIMT group and thickening CIMT group (P < 0.05). The percentage of the left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) took in the thickening CIMT group were both higher than those in the CP group (P < 0.05). Multi-factor logistic regression analysis indicated that there were statistically significant differences in age, homocysteine and left LVMI (P < 0.05). In the Pearson correlation analysis, CIMT and LADI were positively correlated (r= 0.184, P < 0.01) , and there was a positive correlation between CIMT and LVMI (r = 0.236, P < 0.01). Conclusions Left ventricular enlargement is one of the highrisk factors for CIMT abnormalities in patients with acute cerebral infarction. Left atrial and left ventricular enlargement are closely correlated to the severity of CIMT in patients with acute cerebral infarction, indicating that abnormal CIMT in patients with acute cerebral infarction has a certain predictive effect on left atrial and left ventricular enlargement.

To screen the pathogenic mutation location in a genetic family with the neurofibromatosis (NF1) by the next generation sequencing and analyze the clinical phenotype,Illumina Miseq sequencing was applied to capture and analyze the target regions of NF1 family's probands,and furtherly find out the suspicious mutations,as well as to verify the family members by Sanger sequencing.Two rare variants were identified in proband,including the heterozygous missense mutation c.C3649T (p.P1217S) in KIF1B gene and the missense mutation c.T6311C (p.L2104P) on exon 41 of NF1 gene (NM_000267.3).The amino acid at position 2104 was found to be changed from leucine to proline in NF1.The protein prediction SIFT and Polyphen-2 values were 0,0.997,which predicted a conformational change in the encoded protein and eventually affected its function.The mutation c.T6311C in NF1 gene was detected in all patients in this family,which showed genetic co-segregation.The clinical phenotype was neurofibroma in the spinal canal.There were no café au lait spots,iris Lisch nodules,scoliosis,tinnitus,heating loss,or elevated intracranial pressure.The missense mutation c.T6311C (p.L2104P) in NF1 gene might be the genetic cause of this hereditary disease of neurofibromatosis.

This review introduces the system, model, and method of the US ailitary medical logistic supply and support, analyzes the characteristics of the global sourcing and distribution, and highlights the information system development, cen-tral management, efficiency and effective improvement.By learning from the experience of the US Armed Forces, we can be helped to set up new systems and enhance the power of our military medical logistic support.

Objective To evaluate the clinical results of arthroscopic repair of massive rotator cuff tear. Methods The study involved 16 patients with massive rotator cuff tears treated arthroscopically from September 2007 to June 2009. There were 6 males and 11 females at average age 61.5 years (45-75 years). The rotator cuff tears was repaired with arthroscopic double-row reconstruction. The range of motion, pain, strength of flexed elevation and function evaluation score were all recorded before operation and at final follow-up. The results were evaluated by t test and compared according to age and course of disease. Results All patients were healed without complications and the outcome was improved significantly ( P < 0.01 ). The mean VAS score was improved from preoperative 5.6 to postoperative 1.7,the average forward flexion from 69. 1°to 151.2°, the average external rotation from 14.7° to 32.2°, and internal rotation from L1 level to T10, the mean Constant-Murle from 39 to 85, the mean UCLA from 10.4 to 28, the mean SST from 2.8 to 8.8 and the strength of flexed elevation from 10.7% of normal side to 65.0%. Compared with preoperation, there was statistical difference in aspects of pain, range of motion, muscle strength and function in postoperation (P < 0.01 ). Conclusion Arthroscopic doublerow fixation can attain satisfactory results in repair of massive rotator cuff tear.