ObjectiveTo investigate pulmonary function levels and associated influencing factors among rural elderly in Guangzhou， to identify high-risk populations for poor pulmonary function， and to reveal the relationship between the influencing factors of pulmonary function. MethodsWe recruited 1 500 residents aged 60 to 94 years from rural area of Conghua District， Guangzhou City using convenience sampling in 2023. Data on demographics， body measurements， medical history and lifestyle were collected via face-to-face questionnaires and physical examination. Meanwhile， expiratory function parameters including forced expiratory volume in one second （FEV1）， forced vital capacity （FVC）， FEV1/FVC， and the prevalence of airflow obstruction （AFO） were assessed using a portable spirometer. Age and sex distribution of pulmonary function in older adults at 5-year intervals was reported， and risk factors of AFO using multifactorial logistic regression models were analyzed. Furthermore， path analysis was further employed to explore the role of lifestyle in the association between other influencing factors and lung function. ResultsAmong the 1 500 participants， the median age was 71 years （67-75）， and 44.2% were male. Subjects identified as AFOs were generally older， more likely male， less educated， and had lower rates of moderate to vigorous physical activity （＜1 time/week） and lower lean body mass. Mean FEV1/FVC ratio was （82.0±16.4） %. FEV1/FVC was （79.80±17.58） % in men and （83.66±15.22） % in women. Older age， lower education， male sex and leanness were negatively associated with all pulmonary function outcomes （all P values＜0.05）. Path analysis identified that age， gender， marital status， occupation and income may influence pulmonary function indirectly through lifestyle. ConclusionRural elderly in Guangzhou exhibited lower pulmonary function levels， and male sex， non-married status， advanced age， lower education， smoking habits， insufficient engagement in moderate to vigorous physical activity， and lean body type were all associated with worse pulmonary function.

Objective:To establish a rapid quantitative PCR (qPCR) technique for Mycobacterium marinum skin infections, and to analyze its clinical diagnostic efficiency. Methods:DNA was extracted from Mycobacterium marinum colonies and serially diluted (10 -1 to 10 -8). Twelve pairs of previously reported primers and probes, as well as 6 pairs of newly designed primers and probes in this study, were used for qPCR amplification to identify the most sensitive primers and probes for the detection of Mycobacterium marinum. Skin lesion tissues were collected from 72 patients with confirmed Mycobacterium marinum infections (experimental group) and 68 with other mycobacterial infections (control group) at Shandong Provincial Hospital for Skin Diseases & Shandong Provincial Institute of Dermatology and Venereology, Shandong First Medical University & Shandong Academy of Medical Sciences in 2021. These skin tissues were subjected to qPCR amplification, interferon-gamma release assay (IGRA), acid-fast staining, and tissue culture to evaluate the diagnostic efficacy. Results:The newly designed primers and probes targeting the mycobacterial enhanced infection locus 2 (Mel2) demonstrated the highest sensitivity, with a detection limit of 0.86 copies/μl (cycle threshold value = 37) ; the qPCR amplification with the Mel2 primers/probes did not yield positive results when used for the detection of other mycobacteria (including Mycobacterium leprae and Staphylococcus spp) . Among the 72 patients in the experimental group, 44 were positive for qPCR with a sensitivity of 61.1% (95% CI: 49.6% - 71.5%), and 47 were positive for culture with a sensitivity of 65.2% (95% CI: 53.8% - 75.3%) ; all the 68 controls were negative for both qPCR and culture, with their specificities both being 100%. Among 65 patients subjected to IGRA, 31 were positive with a sensitivity of 47.7% (95% CI: 36.0% - 59.6%), while 16 out of 25 controls were negative for IGRA with a specificity of 64.0% (95% CI: 44.5% - 79.8%). Among 58 patients subjected to acid-fast staining, 37 were positive with a sensitivity of 63.8% (95% CI: 50.9% - 74.9%), and 52 out of 66 controls were negative for acid-fast staining with a specificity of 78.8% (95% CI: 67.5% - 86.9%). The combination of qPCR and culture resulted in a sensitivity of 93% and a specificity of 100% for the detection of Mycobacterium marinum. Conclusion:In this study, a highly sensitive qPCR assay was developed for the detection of Mycobacterium marinum, and its combination with culture could further improve the detection sensitivity.

Objective:To investigate the efficacy and safety of oral testosterone therapy in individuals diagnosed with androgen insensitivity syndrome (AIS).Methods:A self-controlled study design was utilized, focusing on individuals with AIS who were genetically diagnosed at the Department of Endocrinology, Genetics, and Metabolism of Beijing Children′s Hospital between 2009 and 2021. These patients underwent treatment involving the administration of testosterone. The primary observed indexes include the measurement of penis length, which should meet the minimal surgical standard (penis length≥2.5 cm) or greater than or equal to -2.5 s (lower limit of normal). Secondary observed indexes include penile length standard deviation score (PL-SDS), an increase in penis longitude (ΔPL), medication dosage, the course of therapy, and safety indicators, among others. There were 4 courses of treatment. After each course, patients were evaluated to determine whether termination of treatment was appropriate. Patients who exhibited inadequate post-treatment penile length growth were advised to continue with further treatment. The statistical methodology included t-test, and a Wilcoxon rank sum test to describe efficacy and safety. The patients were followed up until 2023. Results:The study comprised a total of 51 individuals with AIS, comprising 33 males and 18 females (gender of registered permanent residence). Among these patients, 10 were diagnosed with complete androgen insensitivity syndrome (CAIS) and 41 were diagnosed with partial androgen insensitive syndrome (PAIS). There were 2 children with CAIS were diagnosed by doctors and prescribed testosterone undecanoate, but the children did not really take medicine.The penile length of CAIS patients could not be measured (penile length<0.5 cm) before and after treatment. For PAIS patients, baseline penile length and PL-SDS were (2.3±0.6) cm and -3.7±1.3, respectively. The measurements for penile length and PL-SDS after each treatment course were recorded as follows: (2.7±0.8), (2.8±0.6), (2.6±0.4), (2.6±0.4) cm and -2.8±1.6, 2.5±1.6, 2.9±1.2, -3.2±0.9, respectively. Both penile length and PL-SDS interventions showed statistically significant gains when compared to the baseline performance of the 4 courses ( t=4.05、3.56、2.55、2.23 and 3.88、3.50、2.50、2.19, all P<0.05). Before treatment, 13 PAIS patients (32%) reached 2.5 cm and seven (17%) reached greater than or equal to -2.5 s. Following the initial, subsequent, third, and fourth therapeutic interventions, 18 cases (44%), 24 cases (59%), 25 cases (61%), and 26 cases (63%) reached 2.5 cm, respectively. Additionally, A total of 12 cases (29%), 15 cases (37%), 20 cases (49%), and 21 cases (51%), respectively, were found to reach greater than or equal to -2.5 s. The study involved the longitudinal monitoring of patients with the highest recorded age being 13.7 years. The weight, height, body mass index, bone age/age, cholesterol, hemoglobin and so on were all within the normal range and the difference were not statistically significant (all P>0.05). All 49 patients were no abnormalities in blood electrolyte, liver and kidney function and thyroid function and no changes in precocious puberty, pubic hair growth, aggressive behavior, vulvar skin darkening, diarrhea or other conditions. Conclusions:Testosterone undecanote in children with CAIS was no effective. The initial course of treatment for patients with PAIS demonstrates observable enhancements in penile length and PL-SDS. For patients with inadequate penile length growth, continued treatment in subsequent courses (such as the second, third, and fourth courses) is recommended toenhance outcomes gradually. Testosterone undecanoate was safe and effective for the majority of individuals with PAIS patients, with few adverse effects and good treatment tolerance.

Objective:To explore the dynamic changes of donor derived T cells at different time points in the aplastic anemia mouse model.Methods:The aplastic anemia mouse model was induced and then the proportion of infiltrated donor derived T cells in spleen and bone marrow, expression of activation molecular markers, cell cycle and functional subsets were measured by flow cytometry at different time points to evaluate the functional status of T cells in different periods.Results:①T cell immune-mediated aplastic anemia mouse model was successfully established by half lethal dose irradiation combined with major histocompatibility antigen (MHC) haploidentical lymph node cells infusion. ②The donor derived T cells began to infiltrate significantly in the spleen of aplastic anemia mouse from the 3rd day after transplantation and the ratio of CD4 +/CD8 + gradually inverted. After the 5th day, they gradually entered the bone marrow, predominated by CD8 + cells. ③The expression peak of CD69 in donor CD4 + cells was later than that in CD8 + cells. The trend of CD25 expression in CD4 + cells was the same as that in CD8 + cells, but the expression level in CD8 + cells was higher than CD4 + cells. ④The proportion of donor CD4 + cells in S/G 2/M phase reached the peak in spleen, about 12%, within 3 days after transplantation, while a higher level in CD8 + cells, which was about 20%. And the proportion of both CD4 + and CD8 + cells in S/G 2/M phase increased again after entering bone marrow, which was continued to be higher in CD8 + cells than that in CD4 + cells after 3 days of transplantation. ⑤Immune activated T cells in the spleen rapidly differentiated into effector memory T cells (T EM) after a short central memory T cell (T CM) stage. After entering the bone marrow, some T EM differentiated into effector cells to further function. Conclusion:In the aplastic anemia mouse model, donor derived T cells activated rapidly after entering the allogenic recipient, reached its proliferation booming period and differentiated into T EM cells within 5 days. After 5 days, they began to enter the bone marrow to continue proliferate and damage hematopoiesis.

OBJECTIVE：To improve the quality standard of M ongolian med icine Artemisia sacrorum ，and to provide scientific basis for comprehensive quality evaluation. METHODS ：The appearance and microscopic characteristics of A. sacrorum were identified；scopoletin，chlorogenic acid ，caffeic acid ，scopoletin and 3，5-dicaffeoylquinic acid were identified quantitatively by TLC；the contents of above 5 components were determined by HPLC. The water content ，total ash and extract were examined. RESULTS：The stem of A. sacrorum was cylindrical ，and its surface was purple or purple-brown or cyan-brown ；the leaves were ovate or oblong-ovate ，fragrant；the flowers were yellow ，head-shaped，subglobose or hemispherical. The powder was green or yellow-green，its pollen grain had three germination ；the parenchymal cell clusters with sharp edges and numerous threaded ducts ， occasionally having marginal pitted ducts ；its wood fibers were in bundles mostly. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for 5 substance control and samples. The linear range of scopoletin， chlorogenic acid ， caffeic acid ， scopolactone and 3，5-dicaffeoylquinic acid were 85.60-428.00， 10.16-101.60， 10.20-102.00，40.84-408.40 and 40.80-408.00 μg/mL（all r＞0.999 0）. RSDs of precision ，stability，repeatability tests were all less than 3.00％（n＝6）. The average recoveries were 103.07％，99.66％，98.37％，97.78％，98.40％（all RSDs ＜3.00％，n＝6）. The contents of the above-mentioned 5 compounds in 10 batches of samples were 0.36％-1.23％，0.09％-0.51％，0.04％-0.13％， 0.61％ -1.13％ ，0.12％ -1.11％ ，respectively；the average com contents of water ，total ash and water soluble extract were 6.25％，5.86％，26.50％，respectively. CONCLU SIONS：O the basis of the original quality standard of A. sacrorum ， microscopic identification，TLC identification ，content determination and examination items of water ，total ash and extract are added. The method shows good precision ，accuracy and stability ，which can provide reference for more scientific and standardized evaluation of the quality of this medicinal material.

Objective To detect de novo development of anti-HLA antibodies after renal transplantation, and to investigate their influence on graft function. Methods 384 kidney recipients,who were negative for anti-HLA antibody before transplantation, were monitored for anti-HLA antibodies over a period of 3-96 months, and a sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HLA antibodies. HLA antibody ＞10 % was defined as positive levels. Results Among 384 recipients tested, 318 recipients (82. 8 %) were negative for anti-HLA antibody after transplantation; 66 recipients (17. 2 %) developed de novo HLA antibodies, 3 recipients with HLA class Ⅰ, 61 with HLA class Ⅱ, 2 with both HLA class Ⅰ and Ⅱ. According to amino acid residue matching, 7 cases developed de novo antibodies among 92 recipients with 0 HLA-DR mismatches,compared with 59 cases among 292 recipients with 1-2 mismatches, which showed significant difference between two groups (P＜0. 01 ). 87. 4 % (278/318) recipients negative for HLA antibodies after transplantation achieved good graft function, in comparison with 65. 2 % (43/66) recipients positive for HLA antibodies (P＜0. 05). Conclusion De novo production of HLA antibodies posttransplantation may be closely associated with HLA-DR mismatch. De novo HLA antibodies posttransplantation might damage graft function and reduce graft survival rate. The detection of de novo development of anti-HLA antibodies after renal transplantation has clinical significance for assessing renal allograft function.