To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Objective:To analyze the immunization status of routine vaccines for children aged 0-7 years old with special health status in Tianhe District, Guangzhou City, from 2023 to 2024.Methods:From April 2023 to March 2024, 42 vaccination units in Tianhe District, Guangzhou, were organized to collect data on diseases and vaccination history of children with special health status. Vaccination rates were calculated, and multivariate logistic regression models were used to explore the impact of special health status on vaccination rates.Results:A total of 1 976 children aged 0-7 years old with special health status were included, with an average of (1.26±0.58) diseases per participant. The average number of vaccine doses administered for routine immunizations was (14.29±4.27), and the full vaccination coverage was 72.76%. The vaccination rate of 10 doses in the immunization program vaccine was less than 90.00%. The timely rate of the first dose of hepatitis B vaccine (HepB1) was 74.14%, and that of the first dose of measles vaccine (MCV1) was 63.93%. Compared with children with infectious diseases, those with neonatal diseases were more likely to miss the third dose of poliomyelitis vaccine (PV3), MCV1 and the second dose of Group A meningococcal polysaccharide vaccine (MPSV-A2). Those with neuromuscular system diseases were more likely to miss PV3, MPSV-A2 and the first dose of Japanese encephalitis vaccine, live (JE-L1). Those with congenital heart disease were more likely to miss PV3, the third dose of diphtheria tetanus-acellular pertussis vaccine (DTaP3), MCV1, MPSV-A2, and JE-L1. Those with hematological disorders were more likely to miss PV3, MCV1, MPSV-A2, and JE-L1. Those with genetic diseases were more likely to miss MPSV-A2. Those with comorbidities were more likely to miss MCV1 and MPSV-A2. Those with neonatal diseases, neuromuscular system diseases, congenital heart disease, hematopoietic system diseases, genetic diseases, or comorbidities had difficulties in completing the full vaccination process.Conclusion:Children with special health conditions have lower rates of routine immunization and timely vaccination. More measures are needed to improve vaccination rates.

AIM To investigate the protective effect of Sanfeng Tongqiao Dropping Pills against house dust mite(HDM)-induced allergic asthma in mice.METHODS Compared to the intact BALB/c mice in the blank control group,the BALB/c mice randomly assigned into the model group,the dexamethasone group(0.67 mg/kg),and the low-dose,medium-dose,and high-dose Sanfeng Tongqiao Dropping Pills groups(15,30 and 60 mg/kg),were induced into acute allergic asthma models via weekly intraperitoneal sensitization with 0.1 mL HDM solution(0.5 mg/mL)for three weeks followed by three consecutive daily intranasal challenges with 10 μL HDM solution(2.5 mg/mL)starting in the third week.The drug administered continuously 7 days after the last excitation.The mice had their airway reactive Penh value detected,their pulmonary pathological changes observed by HE staining,their blood eosinophils(EOS)counted,their Th2 cytokines in lung tissue and serum IgE levels detected by ELISA,and their number of peripheral blood mononuclear cells(PBMC)and pulmonary Th2 cells detected by flow cytometry.Chronic allergic asthma was induced in grouped BALB/c mice through repeated intranasal challenges with 10 μL HDM solution(2.5 mg/mL)administered five times weekly for five consecutive weeks.Drug treatment continued for 14 days following the final challenge.After the final treatment,the mice had their pulmonary pathological changes observed by HE staining,and their levels of Th2 cytokines in B ALF and lung tissue and serum IgE detected by ELISA.RESULTS Compared to the blank control group,the acute allergic asthma model group exhibited increases in Penh value,EOS count and IgE level in serum,IL-4 and IL-5 levels in lung tissue(P＜0.01);obvious pulmonary inflammatory cells infiltration,and thickened airway wall;and increase in pulmonary number of Th2 cells(P＜0.01).Compared to the model group,the groups intervened with Sanfeng Tongqiao Dropping Pills demonstrated decreased Penh value,serum EOS count,IgE level and IL-5 level in lung tissue(P＜0.05,P＜0.01);reduced pulmonary inflammatory infiltration and alleviated airway wall thickening;and decreased number of pulmonary Th2 cells.Compared to the blank group,the chronic allergic asthma model group showed obvious pulmonary inflammatory infiltration and airway wall thickening;and increased EOS count and IgE level in serum,IL-4 and IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).Compared to the model group,the groups intervened with either medium-dose or high-dose Sanfeng Tongqiao Dropping Pills demonstrated reduced pulmonary inflammatory infiltration;and decreased serum EOS count,IgE level,IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).CONCLUSION Sanfeng Tongqiao Dropping Pills reduce Th2 cells in peripheral blood and lung tissue,suppress type 2 inflammation,and thereby alleviate allergic asthma.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Objective:To analyze the immunization status of routine vaccines for children aged 0-7 years old with special health status in Tianhe District, Guangzhou City, from 2023 to 2024.Methods:From April 2023 to March 2024, 42 vaccination units in Tianhe District, Guangzhou, were organized to collect data on diseases and vaccination history of children with special health status. Vaccination rates were calculated, and multivariate logistic regression models were used to explore the impact of special health status on vaccination rates.Results:A total of 1 976 children aged 0-7 years old with special health status were included, with an average of (1.26±0.58) diseases per participant. The average number of vaccine doses administered for routine immunizations was (14.29±4.27), and the full vaccination coverage was 72.76%. The vaccination rate of 10 doses in the immunization program vaccine was less than 90.00%. The timely rate of the first dose of hepatitis B vaccine (HepB1) was 74.14%, and that of the first dose of measles vaccine (MCV1) was 63.93%. Compared with children with infectious diseases, those with neonatal diseases were more likely to miss the third dose of poliomyelitis vaccine (PV3), MCV1 and the second dose of Group A meningococcal polysaccharide vaccine (MPSV-A2). Those with neuromuscular system diseases were more likely to miss PV3, MPSV-A2 and the first dose of Japanese encephalitis vaccine, live (JE-L1). Those with congenital heart disease were more likely to miss PV3, the third dose of diphtheria tetanus-acellular pertussis vaccine (DTaP3), MCV1, MPSV-A2, and JE-L1. Those with hematological disorders were more likely to miss PV3, MCV1, MPSV-A2, and JE-L1. Those with genetic diseases were more likely to miss MPSV-A2. Those with comorbidities were more likely to miss MCV1 and MPSV-A2. Those with neonatal diseases, neuromuscular system diseases, congenital heart disease, hematopoietic system diseases, genetic diseases, or comorbidities had difficulties in completing the full vaccination process.Conclusion:Children with special health conditions have lower rates of routine immunization and timely vaccination. More measures are needed to improve vaccination rates.

AIM To investigate the protective effect of Sanfeng Tongqiao Dropping Pills against house dust mite(HDM)-induced allergic asthma in mice.METHODS Compared to the intact BALB/c mice in the blank control group,the BALB/c mice randomly assigned into the model group,the dexamethasone group(0.67 mg/kg),and the low-dose,medium-dose,and high-dose Sanfeng Tongqiao Dropping Pills groups(15,30 and 60 mg/kg),were induced into acute allergic asthma models via weekly intraperitoneal sensitization with 0.1 mL HDM solution(0.5 mg/mL)for three weeks followed by three consecutive daily intranasal challenges with 10 μL HDM solution(2.5 mg/mL)starting in the third week.The drug administered continuously 7 days after the last excitation.The mice had their airway reactive Penh value detected,their pulmonary pathological changes observed by HE staining,their blood eosinophils(EOS)counted,their Th2 cytokines in lung tissue and serum IgE levels detected by ELISA,and their number of peripheral blood mononuclear cells(PBMC)and pulmonary Th2 cells detected by flow cytometry.Chronic allergic asthma was induced in grouped BALB/c mice through repeated intranasal challenges with 10 μL HDM solution(2.5 mg/mL)administered five times weekly for five consecutive weeks.Drug treatment continued for 14 days following the final challenge.After the final treatment,the mice had their pulmonary pathological changes observed by HE staining,and their levels of Th2 cytokines in B ALF and lung tissue and serum IgE detected by ELISA.RESULTS Compared to the blank control group,the acute allergic asthma model group exhibited increases in Penh value,EOS count and IgE level in serum,IL-4 and IL-5 levels in lung tissue(P＜0.01);obvious pulmonary inflammatory cells infiltration,and thickened airway wall;and increase in pulmonary number of Th2 cells(P＜0.01).Compared to the model group,the groups intervened with Sanfeng Tongqiao Dropping Pills demonstrated decreased Penh value,serum EOS count,IgE level and IL-5 level in lung tissue(P＜0.05,P＜0.01);reduced pulmonary inflammatory infiltration and alleviated airway wall thickening;and decreased number of pulmonary Th2 cells.Compared to the blank group,the chronic allergic asthma model group showed obvious pulmonary inflammatory infiltration and airway wall thickening;and increased EOS count and IgE level in serum,IL-4 and IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).Compared to the model group,the groups intervened with either medium-dose or high-dose Sanfeng Tongqiao Dropping Pills demonstrated reduced pulmonary inflammatory infiltration;and decreased serum EOS count,IgE level,IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).CONCLUSION Sanfeng Tongqiao Dropping Pills reduce Th2 cells in peripheral blood and lung tissue,suppress type 2 inflammation,and thereby alleviate allergic asthma.

Objective:To evaluate the efficacy and safety of PD-1 inhibitor combined with azacitidine and HAG regimen in the treatment of relapsed/refractory acute myeloid leukemia(R/R AML).Methods:This study is a prospective,single-arm,phase Ⅱ clinical trial that included R/R AML patients who met the inclusion criteria and were treated at The First Affiliated Hospital of Soochow University from December 2020 to August 2023.Patients could undergo allogeneic hematopoietic stem cell transplantation(allo-HSCT)after salvage therapy.The efficacy and safety were evaluated.Results:Twenty patients were enrolled,including 14 males and 6 females,with an average age of(50.7±15.3)years.The overall response rate(ORR)after one cycle of the treatment was 75.0％(15/20),and 35.0％(7/20)of the patients achieved complete remission(CR)or complete remission with incomplete hematologic recovery(CRi)after two cycles of the treatment.Eight patients received allo-HSCT.The main adverse events were hematologic toxicities,and no grade 5 adverse events occurred.Conclusion:The combination of PD-1 inhibitor,azacitidine,and the HAG regimen is a feasible and relatively safe treatment option for R/R AML,thus,to be worth further study.

Objective:To investigate the changes of cerebral blood flow before and after transfusion in neonates with anemia,and analyze the related influencing factors of neonatal middle cerebral artery blood flow.Methods:A total of 39 neonates with anemia who were hospitalized in the Department of Neonatology of Shaanxi Provincial People's Hospital from October 2021 to March 2023 and received blood transfusion treatment were selected.Basic data were collected.Transcranial Doppler ultrasound was used to collect peak systolic flow velocity(Vs),end-diastolic flow velocity(Vd)and vascular resistance index(RI)of left and right middle cerebral artery(MCA).To analyze the relationship between neonatal anemia and middle cerebral artery blood flow velocity.To explore the effects of anemia and blood transfusion on middle cerebral artery blood flow,and analyze the related factors of middle cerebral artery blood flow.Results:A total of 39 neonates were included in this study,and the Vs on both the left and right sides of the middle cerebral artery after transfusion was lower than that before transfusion[Vs on the left side after transfusion was(44.7±16.7)cm/s compared with that before transfusion(45.9±19.2)cm/s,Vs on the right side after transfusion:(49.2±18.4)cm/s Vs(52.4±25.1)cm/s before transfusion];The mean blood pressure,Vd and RI after transfusion were all higher than those before transfusion[mean pressure(after transfusion/before transfusion):(41.7±6.3)mmHg ratio(40.9±6.9)mmHg],[Vd after transfusion on the left side:(11.7±6.6)cm/s compared with that before transfusion(10.9±5.0)cm/s,Vd after transfusion on the right side:(10.5±4.0)cm/s compared with(9.6±5.5)cm/s],[left post-transfusion RI:(0.75±0.08)compared with pre-transfusion RI:(0.74±0.09),right post-transfusion RI:(0.77±0.08)compared with(0.70±0.86)before transfusion],but the difference was not statistically significant(P＞0.05);Through further correlation analysis,gestational age at birth,standard deviation of hemoglobin and normal value before and after transfusion,mean blood pressure,birth weight and blood flow of middle cerebral artery were respectively correlated,and it was found that gestational age was positively correlated with MCA Vd before transfusion,the standard deviation of hemoglobin before transfusion was negatively correlated with MCA on the left and right side,and the mean blood pressure was positively correlated with MCA blood flow.Birth weight was positively correlated with right side Vd of MCA after transfusion.Conclusion:Blood transfusion in anemic neonates can affect blood flow velocity in middle cerebral artery.The blood flow velocity of middle cerebral artery was correlated with gestational age,anemia degree,mean blood pressure and birth weight.

To formulate an expert consensus on the principles of preoperative hair removal and provide scientific guidance for standardized removal of hair before surgical procedures so as to reduce the incidence of surgical site infections.METHODS Led by the Hospital Management Institute of National Health Commission of the People's Republic of China,this consensus was reached with the joint efforts from the expects of relevant fields such as surgeries,interventional therapies,nursing,and infection prevention and control.The consensus facilitates the classification and evaluation of literatures by following the evidence grade formulated by Oxford Evidence-based Medicine Center and focuses on the association of preoperative hair removal with surgical site infection,it reaches the evidence grade of expert consensus and recommendation intensity by integrating with discussions on meetings and clinical experience of the expects from relevant fields.RESULTS A total of 6 items of consensus were reached by summarizing the latest evidence on the aspects including the indications for preoperative hair removal,tools,range,timing and places.CONCLUSION The consensus,to some extent,make supplements to and complete the exiting regulations and standards.It provides guidance for the medical institutions to carry out the preoperative hair removal.

Objective To report the diagnosis,treatment,and verification process of a patient with sex development disorder whose chromosomal karyotype and genetic test results are inconsistent,and conduct a literature review to improve the understanding of the mosaic status of sexual development disorders.Methods A 14-year-old patient presented with primary amenorrhea on April 3,2020,at the First Affiliated Hospital of Hebei North University,exhibiting female sexual characteristics.The patient underwent ultrasonic/magnetic resonance imaging of gonads,assessment of gonadal axis function,chromosomal karyotype,and molecular genetic testing,as well as pelvic exploration,malignant gonads resection,and hormone replacement therapy,resulting in drug-induced menstruation.During the diagnosis and treatment,it was found that the patient's chromosomal karyotype analysis was inconsistent with the molecular genetic test results.Subsequently,samples from the three germ layer cells were taken,and fluorescence in situ hybridization(FISH)was used to detect the sex chromosomes in each germ layer cell.XY probes were used to label the gonadal pathological sections to explore the distribution differences of the Y chromosome in the gonads,and changes in anti-Müllerian hormone(AMH)levels before and after surgery were compared.Databases such as Wanfang and PubMed were searched to summarize relevant cohort study literature and understand the current status of research on this disease.Results The patient's body exhibited a significant differences between the 45,X and 46,XY cell lines in different germ layers and within the same layer tissues.The proportion of 45,X in buccal mucosal cells derived from the ectoderm was 30%(6/20),in peripheral blood lymphocytes derived from the mesoderm was 9.7%(11/114),and in bladder shed cells derived from endoderm was 20.4%(22/108).The gonadal pathological sections labeled with XY probes indicated a mosaic state with a reduced Y-chromosome;where the epididymal structure area had a 45,X cell line mosaic of 50.0%,and the malignant area had a normal"Y"content.After gonadal resection,AMH levels significantly dropped from 7.28 pmol/L to＜0.07 pmol/L.Literature review revealed that patients with 45,X/46,XY have a complex phenotype spectrum,most with features of Turner syndrome,and female phenotypes are at risk of gonadal tumors.Conclusions In the diagnosis of difficult cases of sex development disorders,when performing peripheral blood karyotype testing,the number of counted cells and analyzed cells should be increased as much as possible,and multi-germ layer cell sampling should be performed.Gonads with a high"Y"mosaic rate are more prone to malignancy in the abdominal cavity.Detecting AMH levels can distinguish cryptorchidism and anorchidism in sexual development disorders with Y chromosomes.