Objective:To investigate the expression levels, correlation, and diagnostic efficacy of peripheral blood CD8 +T lymphocyte subsets and different immune checkpoint markers in patients with Brucellosis. Method:A case-control study was conducted on 32 patients with acute phase brucellosis (27 males and 5 females, aged 36 (30, 43) years), 38 patients with chronic phase brucellosis (30 males and 8 females, aged 40 (32, 48) years), and 30 healthy controls (24 males and 6 females, aged 39 (32, 46) years), who underwent physical examination at Xinjiang Uygur Autonomous Region People′s Hospital from February 1, 2021 to December 31, 2023. All subjects had fasting blood sampling once in the morning. Flow cytometry was used to detect the proportion of peripheral blood lymphocyte subsets, the expression levels of CD8 +T lymphocyte surface programmed cell death receptor-1 (PD-1), T lymphocyte immunoglobulin receptor with Ig and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain containing protein 3 (TIM-3), perforin and granzyme B. The changes in these indicators during the acute and chronic phases of the disease were observed, and correlation analysis was performed using Spearman′s method. Receiver Operating Characteristic Curve (ROC) analysis is used to evaluate the diagnostic value of immunological indicators with differences in acute and chronic brucellosis. Results:CD3 +T lymphocyte in the chronic group (70.71%±8.78%) is significantly lower than that in the healthy control group (74.65%±7.31%) ( P<0.05), and CD4 +T lymphocyte in the acute phase group (39.52%±5.85%) is also lower than that in the healthy control group (45.10%±5.18%) ( P<0.01); while CD8 +T lymphocyte in the acute group (31.73%±5.87%) is significantly higher than that in the chronic phase group (26.75%±4.71%) ( P<0.001). There was a statistically significant difference ( P<0.001) in CD8+CD28 -T lymphocyte among the acute group (69.85% (58.62%, 78.55%)), chronic group (86.46% (73.30%, 92.52%)) and healthy control group (25.39% (20.60%,32.90%)), when compared pairwise. The expression levels of immune checkpoint PD-1, TIGIT, and TIM-3 on the surface of CD8 +T lymphocytes were higher in both the acute and chronic groups than in the healthy control group ( P<0.001). The expression level of perforin secreted by CD8 +T lymphocytes in the acute and chronic groups was lower than that in the healthy control group ( P<0.05), while the expression level of granzyme B in the acute and chronic groups was higher than that in the healthy control group ( P<0.01). The proportion of CD8 +CD28 -T lymphocytes in brucellosis patients was positively correlated with the expression levels of TIGIT and TIM-3 ( r=0.624, 0.406, P<0.001). The ROC curve combined with the proportion of CD8 +CD28 -T lymphocytes and the proportion of TIGIT on the surface of CD8 +T lymphocytes can distinguish acute and chronic brucellosis. The Area Under Curve (AUC) is 0.973, which has certain implications for clinical differentiation of patients with acute and chronic diseases. Conclusion:The CD8 +T lymphocyte subsets in patients with brucellosis exhibit dynamic changes, accompanied by changes in relevant immune checkpoint molecules, and can regulate the activation and inhibition of the immune status of brucellosis patients. The synergistic effect of CD8 +CD28 -T cells and TIGIT/TIM-3 may be a key mechanism of driving chronicity, and their combined diagnosis can serve as a clinical staging marker.

Objective:To investigate the expression levels, correlation, and diagnostic efficacy of peripheral blood CD8 +T lymphocyte subsets and different immune checkpoint markers in patients with Brucellosis. Method:A case-control study was conducted on 32 patients with acute phase brucellosis (27 males and 5 females, aged 36 (30, 43) years), 38 patients with chronic phase brucellosis (30 males and 8 females, aged 40 (32, 48) years), and 30 healthy controls (24 males and 6 females, aged 39 (32, 46) years), who underwent physical examination at Xinjiang Uygur Autonomous Region People′s Hospital from February 1, 2021 to December 31, 2023. All subjects had fasting blood sampling once in the morning. Flow cytometry was used to detect the proportion of peripheral blood lymphocyte subsets, the expression levels of CD8 +T lymphocyte surface programmed cell death receptor-1 (PD-1), T lymphocyte immunoglobulin receptor with Ig and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain containing protein 3 (TIM-3), perforin and granzyme B. The changes in these indicators during the acute and chronic phases of the disease were observed, and correlation analysis was performed using Spearman′s method. Receiver Operating Characteristic Curve (ROC) analysis is used to evaluate the diagnostic value of immunological indicators with differences in acute and chronic brucellosis. Results:CD3 +T lymphocyte in the chronic group (70.71%±8.78%) is significantly lower than that in the healthy control group (74.65%±7.31%) ( P<0.05), and CD4 +T lymphocyte in the acute phase group (39.52%±5.85%) is also lower than that in the healthy control group (45.10%±5.18%) ( P<0.01); while CD8 +T lymphocyte in the acute group (31.73%±5.87%) is significantly higher than that in the chronic phase group (26.75%±4.71%) ( P<0.001). There was a statistically significant difference ( P<0.001) in CD8+CD28 -T lymphocyte among the acute group (69.85% (58.62%, 78.55%)), chronic group (86.46% (73.30%, 92.52%)) and healthy control group (25.39% (20.60%,32.90%)), when compared pairwise. The expression levels of immune checkpoint PD-1, TIGIT, and TIM-3 on the surface of CD8 +T lymphocytes were higher in both the acute and chronic groups than in the healthy control group ( P<0.001). The expression level of perforin secreted by CD8 +T lymphocytes in the acute and chronic groups was lower than that in the healthy control group ( P<0.05), while the expression level of granzyme B in the acute and chronic groups was higher than that in the healthy control group ( P<0.01). The proportion of CD8 +CD28 -T lymphocytes in brucellosis patients was positively correlated with the expression levels of TIGIT and TIM-3 ( r=0.624, 0.406, P<0.001). The ROC curve combined with the proportion of CD8 +CD28 -T lymphocytes and the proportion of TIGIT on the surface of CD8 +T lymphocytes can distinguish acute and chronic brucellosis. The Area Under Curve (AUC) is 0.973, which has certain implications for clinical differentiation of patients with acute and chronic diseases. Conclusion:The CD8 +T lymphocyte subsets in patients with brucellosis exhibit dynamic changes, accompanied by changes in relevant immune checkpoint molecules, and can regulate the activation and inhibition of the immune status of brucellosis patients. The synergistic effect of CD8 +CD28 -T cells and TIGIT/TIM-3 may be a key mechanism of driving chronicity, and their combined diagnosis can serve as a clinical staging marker.

Objective To investigate the risk factors of subdelirium syndrome in adult ICU patients,then establish a risk prediction model and have it verified.Method A total of 380 adult ICU patients in our hospital between June 2022 and January 2024 were selected in this study.Among the patients,224(70％)were assigned to the model group and 114(30％)to the validation group.Independent risk factors were screened by comparison of the general data,disease and treatment,laboratory indicators and other relevant data between the patients with and without subdelirium.A risk prediction model was established with Logistic regression.Results A risk prediction model was finalised and established.It composed six risk factors of age(OR=1.023),APACHE Ⅱ score(OR=1.093),critical care pain observation tool(OR=2.216),duration of mechanical ventilation(OR=1.003),constraint(OR=2.615)and sepsis(OR=2.081).In internal validation,it was found that the calibration curve closely overlapped with the ideal curve and the area under the ROC curve was 0.816,with a predictive cut off value of 85 points.In external validation,the calibration curve was found closely overlapped with the ideal curve and the area under ROC curve was 0.808.Conclusion The risk prediction model for subdelirium syndrome in adult ICU patients established in the study has good consistency and prediction efficiency,thereby it provides a basis for early clinical screening and intervention of subdelirium syndrome.

Objective To investigate the risk factors of subdelirium syndrome in adult ICU patients,then establish a risk prediction model and have it verified.Method A total of 380 adult ICU patients in our hospital between June 2022 and January 2024 were selected in this study.Among the patients,224(70％)were assigned to the model group and 114(30％)to the validation group.Independent risk factors were screened by comparison of the general data,disease and treatment,laboratory indicators and other relevant data between the patients with and without subdelirium.A risk prediction model was established with Logistic regression.Results A risk prediction model was finalised and established.It composed six risk factors of age(OR=1.023),APACHE Ⅱ score(OR=1.093),critical care pain observation tool(OR=2.216),duration of mechanical ventilation(OR=1.003),constraint(OR=2.615)and sepsis(OR=2.081).In internal validation,it was found that the calibration curve closely overlapped with the ideal curve and the area under the ROC curve was 0.816,with a predictive cut off value of 85 points.In external validation,the calibration curve was found closely overlapped with the ideal curve and the area under ROC curve was 0.808.Conclusion The risk prediction model for subdelirium syndrome in adult ICU patients established in the study has good consistency and prediction efficiency,thereby it provides a basis for early clinical screening and intervention of subdelirium syndrome.

Objective To evaluate the clinical value of neutrophil CD64(nCD64)index as a novel biomark-er in the differential diagnosis of acute and chronic brucellosis.Methods A total of 38 patients with acute bru-cellosis and 48 patients with chronic brucellosis diagnosed in the People's Hospital of Xinjiang Uygur Autono-mous Region from February 2021 to July 2023 were included.Peripheral blood of the patients was collected and nCD64 index was detected by flow cytometry,and the correlation between nCD64 index and disease severi-ty was analyzed.Receiver operating characteristic(ROC)curve was used to analyze the sensitivity and the specificity of nCD64 index in differentially diagnosing acute and chronic brucellosis.Meanwhile,Rose-Bengal Plate Test(RBPT)and Standard Tube Agglutination Test(SAT)were used as controls to evaluate the clini-cal diagnostic value of the three.Results The nCD64 index of acute brucellosis patients was higher than that of chronic brucellosis patients(U=216.00,P<0.001),and the index was positively correlated with the sever-ity of the disease(r=0.670,P<0.001).The ROC curve analysis results showed that the area under the curve of nCD64 index in the differential diagnosis of acute and chronic brucellosis was 0.882(95%CI:0.811-0.952,P<0.001),the cut-off value was 2.81,and sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 83.3%,81.6%,80.4%,81.9%and 82.6%,respectively.The efficacy of nCD64 index in differential diagnosis of nCD64 index was significantly better than those of the qualitative tests of RBPT and SAT.Conclusion nCD64 index has favourable sensitivity and specificity in the differential diag-nosis of acute and chronic brucellosis,and tends to reflect the severity of the disease.It has clinical value in the differential diagnosis of acute brucellosis and chronic brucellosis,and plays an important role in the early diag-nosis and treatment effect monitoring of brucellosis.

Objective To investigate the characteristics of changes in interleukin(IL)-33 and regulatory T(Treg)cells in brucellosis,to verify the regulatory effect of IL-33 on Treg cells,so as to clarify the immune mechanism of IL-33 on Treg cells in brucellosis.Methods The peripheral blood of 39 patients with brucellosis treated in the People's Hospital of Xinjiang Uygur Autonomous Region from January to December 2021(the brucellosis group)and 42 healthy controls(the healthy control group)who underwent physical examination during the same period were collected.The serum IL-33 level was detected by AimPlex kit,and the proportion of Treg cells was detected by flow cytometry.Peripheral blood mononuclear cell(PBMC)was extracted and cultured in vitro to observe the proportion and mRNA expression levels of forkhead box protein P3(Foxp3)after stimulation and blocking of IL-33.Results Compared with the healthy control group,the level of IL-33 and the proportion of Treg cells in brucellosis group were significantly increased,with statistical significance(P＜0.05).In vitro tests showed that the Foxp3 proportion and mRNA expression level of PBMC in the two groups were significantly increased after IL-33 stimulation,and significantly decreased after IL-33 blocking,with statistical significance(P＜0.001).Conclusion IL-33 and Treg cells increased significantly in brucellosis patients,and IL-33 promoted the immune function of Treg cells.Blocking IL-33 is expected to be a potential target for immunotherapy of brucellosis.

【Objective】 To investigate the development of adaptability in children aged 2 - 6, and to explore its influencing factors, so as to provide reference for promoting the development of adaptability in young children. 【Methods】 Data were from the National Nutrition and Health Systematic Survey for Children in China, and 3 319 children aged 2 - 6 and their parents from 28 sites across 14 provinces were recruited in this study.The Development Scale for Children Aged 0 - 6 years (WS/T 580-2017) was used to measure the developmental quotient of children′s adaptive ability, and a survey questionnaire was used to collect relevant information about children and their parents. 【Results】 Among 3 319 children aged 2 - 6, the proportion of slightly low or low level of adaptability, moderate adaptability development, good and excellent adaptability development was 7.68%,66.25% and 26.06%, respectively.The proportion of children aged 5 - 6 with good and excellent adaptability was lower in 3-year-old and 4-year-old groups (χ²=59.29, P<0.05).Multiple stepwise linear regression showed that children′s gender (β=0.06), gestational age of birth (β=-0.05), only child (β=-0.04), left-behind child (β=-0.04), the main caregiver (β=-0.06), and the education level of parents (β=0.09, 0.10), whether parents actively pay attention to children′s emotions (β=-0.06) and whether children play with homemade toys (β=-0.04) were the influencing factors of children′s adaptive development quotient.Girls, full-term children, only children, non-left-behind children, children with parents as main caregivers, parents with a high level of education, parents who often take the initiative to pay attention to children′s emotions, and children who play with homemade toys had a higher level of adaptability development quotient. 【Conclusions】 The development level of adaptability in children aged 2 - 6 in China is mostly above the average level and is related to multiple factors.Targeted intervention work can be carried out on relevant factors in order to promote the development of children′s adaptability.

Objective:The aim is to understand the current situation of death literacy among Chinese residents in the Guangdong-Hongkong-Macao Greater Bay Area and explore educational approaches to enhance the death literacy of the social public.Method:The Death Literacy Index was conducted among 2,002 residents in the Guangdong-Hong Kong-Macao Greater Bay Area,which included 29 items from four dimensions namely practice,experience,fact,and community.Results:The average score of death literacy among the respondents was moderate(6.70),with scores for practical and experiential knowledge(7.32,7.50)higher than those for factual and community knowledge(6.20,6.10).The weakest content included discussing bereavement with people who have just lost loved ones(5.07),laws and regulations related to death at home(5.35),and providing support for the terminal person and their families in the community.Conclusion:The death literacy of the Chinese residents in the Greater Bay Area was moderate,in which community knowledge was the weakest with a lack of concern and support for the terminal person and the caregivers.Therefore,community health promotion strategies were proposed to strengthen death education for the public and promote the death literacy of the residents.

Objective:To explore the clinical application value of multiple cytogenetic and molecular genetic techniques in the diagnosing of chromosomal microstructural abnormalities.Methods:The proband was a 30-year-old man. He went to Reproductive Medicine Center of Shenzhen Zhongshan Urology Hospital in July 2021 because of a 7-year history of primary infertility. Chromosome karyotype was analyzed by conventional G-banding technique. One case was found to be suspected of microstructural aberration in X chromosome. The origin and structural characteristics of this X chromosome structural aberration was identified by high-resolution G-banding, copy number variation sequencing (CNV-seq), C-banding and fluorescence in situ hybridization (FISH). The pregnancy outcome of this case was followed up. Results:Conventional G-banding karyotype of peripheral blood lymphocytes was initially diagnosed as 46, Y, ?inv(X)(p22.3p22.2). The final karyotype of proband was interpreted as 46, Y, der(X) t(X; Y)(p22.3; q12) mat by high-resolution G-banding karyotype analysis, CNV-seq, C-banding analysis and FISH detection. His spouse had conceived singleton pregnancy via in vitro fertilization and embryo transfer. Prenatal diagnosis had been performed. Karyotype of amniotic fluid was 46, X, der(X) t(X; Y)(p22.3; q12) pat. No structural malformation was detected prenatally by ultrasound. The neonate was phenotypically normal one month after birth. Conclusion:The combined application of multiple cytogenetic and molecular genetic techniques can provide a reliable technical platform for characterizing the microscopic structural aberrations of chromosomes, and an important genetic basis for exploring the phenotype, prognosis and offspring risk of such patients.

Objective:To explore the clinical application value of multiple cytogenetic and molecular genetic techniques in the diagnosing of chromosomal microstructural abnormalities.Methods:The proband was a 30-year-old man. He went to Reproductive Medicine Center of Shenzhen Zhongshan Urology Hospital in July 2021 because of a 7-year history of primary infertility. Chromosome karyotype was analyzed by conventional G-banding technique. One case was found to be suspected of microstructural aberration in X chromosome. The origin and structural characteristics of this X chromosome structural aberration was identified by high-resolution G-banding, copy number variation sequencing (CNV-seq), C-banding and fluorescence in situ hybridization (FISH). The pregnancy outcome of this case was followed up. Results:Conventional G-banding karyotype of peripheral blood lymphocytes was initially diagnosed as 46, Y, ?inv(X)(p22.3p22.2). The final karyotype of proband was interpreted as 46, Y, der(X) t(X; Y)(p22.3; q12) mat by high-resolution G-banding karyotype analysis, CNV-seq, C-banding analysis and FISH detection. His spouse had conceived singleton pregnancy via in vitro fertilization and embryo transfer. Prenatal diagnosis had been performed. Karyotype of amniotic fluid was 46, X, der(X) t(X; Y)(p22.3; q12) pat. No structural malformation was detected prenatally by ultrasound. The neonate was phenotypically normal one month after birth. Conclusion:The combined application of multiple cytogenetic and molecular genetic techniques can provide a reliable technical platform for characterizing the microscopic structural aberrations of chromosomes, and an important genetic basis for exploring the phenotype, prognosis and offspring risk of such patients.