Objective:To establish a flow cytometric assay for myeloid-derived suppressor cells (MDSC) in human peripheral whole blood and reference intervals for healthy adults in Shanghai region.Method:A whole blood eight-color and twelve-parameter flow cytometric assay was designed, utilizing fluorescently labeled antibodies against CD45, CD3, CD19, CD123, CD56, CD16, HLA-DR, CD33, CD11b, CD14, CD15 and CD20.A total of 246 healthy participants who met the health standards from the health check-ups conducted at the Tongji University Affiliated Shanghai East Hospital between May 8 to December 2, 2024 were enrolled. Peripheral venous whole blood was collected using EDTA-K 2 anticoagulant vacuum tubes for MDSC detection. A single-platform flow cytometry based relative count technique was used to quantify the percentage of each MDSC subpopulation. Kolmogorov Smirnov (K-S) test was used to test the distribution of specimens. Mann-Whitney U test and Kruskal-Wallis (K-W) test were used to evaluate whether reference intervals should be established separately based on gender or age. According to the clinical significance of MDSC, bilateral reference intervals were taken. Non parametric methods were used to take the percentile P2.5 and P97.5 to represent the rank of the lower and upper reference limits, respectively. Results:The results showed that a gating strategy was designed to exclude granulocytes, lymphocyte lineage cells, and natural killer cells. The K-S test results showed that the MDSC in each group of healthy individuals were distributed in a skewed manner. The U test showed significant gender differences ( P0.05) in the distribution of total myeloid-derived suppressor cells (T-MDSC) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). The K-W test showed no significant differences in MDSC among different age groups (21-30 years old, 30-40 years old, 40-50 years old, and 50-60 years old). T-MDSC reference interval is 0.056%-0.485%, PMN-MDSC reference interval is 0.035%-0.406%, Monocytic myeloid-derived suppressor cells (M-MDSC) reference interval is 0.000%-0.221%, early-stage myeloid-derived suppressor cells (E-MDSC) reference interval is 0.004%-0.125%. Reference interval verification was conducted on 20 healthy individuals, with a pass rate of 100%. Conclusion:A whole blood eight-color and twelve-parameter flow cytometric assay was established in this experiment. Based on the flow cytometry single platform method, reference intervals for healthy adults in Shanghai region were established.

Objective:To establish a flow cytometric assay for myeloid-derived suppressor cells (MDSC) in human peripheral whole blood and reference intervals for healthy adults in Shanghai region.Method:A whole blood eight-color and twelve-parameter flow cytometric assay was designed, utilizing fluorescently labeled antibodies against CD45, CD3, CD19, CD123, CD56, CD16, HLA-DR, CD33, CD11b, CD14, CD15 and CD20.A total of 246 healthy participants who met the health standards from the health check-ups conducted at the Tongji University Affiliated Shanghai East Hospital between May 8 to December 2, 2024 were enrolled. Peripheral venous whole blood was collected using EDTA-K 2 anticoagulant vacuum tubes for MDSC detection. A single-platform flow cytometry based relative count technique was used to quantify the percentage of each MDSC subpopulation. Kolmogorov Smirnov (K-S) test was used to test the distribution of specimens. Mann-Whitney U test and Kruskal-Wallis (K-W) test were used to evaluate whether reference intervals should be established separately based on gender or age. According to the clinical significance of MDSC, bilateral reference intervals were taken. Non parametric methods were used to take the percentile P2.5 and P97.5 to represent the rank of the lower and upper reference limits, respectively. Results:The results showed that a gating strategy was designed to exclude granulocytes, lymphocyte lineage cells, and natural killer cells. The K-S test results showed that the MDSC in each group of healthy individuals were distributed in a skewed manner. The U test showed significant gender differences ( P0.05) in the distribution of total myeloid-derived suppressor cells (T-MDSC) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). The K-W test showed no significant differences in MDSC among different age groups (21-30 years old, 30-40 years old, 40-50 years old, and 50-60 years old). T-MDSC reference interval is 0.056%-0.485%, PMN-MDSC reference interval is 0.035%-0.406%, Monocytic myeloid-derived suppressor cells (M-MDSC) reference interval is 0.000%-0.221%, early-stage myeloid-derived suppressor cells (E-MDSC) reference interval is 0.004%-0.125%. Reference interval verification was conducted on 20 healthy individuals, with a pass rate of 100%. Conclusion:A whole blood eight-color and twelve-parameter flow cytometric assay was established in this experiment. Based on the flow cytometry single platform method, reference intervals for healthy adults in Shanghai region were established.

Objective: To explore whether the survival benefit of the third-line extensive small-cell lung cancer (ES-SCLC) will be obtained by the combination of anlotinib and radiotherapy, and evaluate the safety of this treatment regimen. Methods: Twenty-seven patients with ES-SCLC who received third-line treatment with less than three metastatic organs at the Cancer Hospital of Xinjiang Medical University from November 2018 to July 2021 were collected and treated with radiotherapy based on anlotinib. Kaplan-Meier curve was used to estimate the overall survival (OS) and progression-free survival (PFS), descriptive statistical analysis was used to evaluate the safety, and European organisation for research and treatment of cancer quality of life questionnaire-core 30 (EORTC QLQ-C30) was used to evaluate the quality of life. Results: The follow-up cut-off date was July 1, 2021, and the follow-up time ranged from 4.8 to 31.0 months, with a median follow-up time of 10.2 months for the entire group. Among the 27 patients, 4 achieved partial remission, 17 had stable disease and 6 had progression of disease. The objective remission rate (ORR) was 14.8%, and the disease control rate (DCR) was 77.8%. Median PFS and the median OS were 5 months and 11 months, respectively. The most common adverse reactions included fatigue (33.3%, 9/27), anorexia (14.8%, 4/27), bleeding (14.8%, 4/27) and hand-foot syndrome (11.1%, 3/27). Most of them were grade 1 to grade 2, 3 cases were more than grade 3, and there was no grade 5 toxicity recorded. After radiotherapy combined with amlotinib treatment, patients showed improvement in general health, somatic functioning, social functioning, and emotional functioning (all P<0.05). Conclusion: For the third-line ES-SCLC patients, radiotherapy based on the anlotinib can significantly prolong their PFS and OS, and the adverse reactions can be tolerated.
Humans ; Small Cell Lung Carcinoma/radiotherapy* ; Lung Neoplasms/radiotherapy* ; Quality of Life ; Treatment Outcome ; Progression-Free Survival

Objective: To explore whether the survival benefit of the third-line extensive small-cell lung cancer (ES-SCLC) will be obtained by the combination of anlotinib and radiotherapy, and evaluate the safety of this treatment regimen. Methods: Twenty-seven patients with ES-SCLC who received third-line treatment with less than three metastatic organs at the Cancer Hospital of Xinjiang Medical University from November 2018 to July 2021 were collected and treated with radiotherapy based on anlotinib. Kaplan-Meier curve was used to estimate the overall survival (OS) and progression-free survival (PFS), descriptive statistical analysis was used to evaluate the safety, and European organisation for research and treatment of cancer quality of life questionnaire-core 30 (EORTC QLQ-C30) was used to evaluate the quality of life. Results: The follow-up cut-off date was July 1, 2021, and the follow-up time ranged from 4.8 to 31.0 months, with a median follow-up time of 10.2 months for the entire group. Among the 27 patients, 4 achieved partial remission, 17 had stable disease and 6 had progression of disease. The objective remission rate (ORR) was 14.8%, and the disease control rate (DCR) was 77.8%. Median PFS and the median OS were 5 months and 11 months, respectively. The most common adverse reactions included fatigue (33.3%, 9/27), anorexia (14.8%, 4/27), bleeding (14.8%, 4/27) and hand-foot syndrome (11.1%, 3/27). Most of them were grade 1 to grade 2, 3 cases were more than grade 3, and there was no grade 5 toxicity recorded. After radiotherapy combined with amlotinib treatment, patients showed improvement in general health, somatic functioning, social functioning, and emotional functioning (all P<0.05). Conclusion: For the third-line ES-SCLC patients, radiotherapy based on the anlotinib can significantly prolong their PFS and OS, and the adverse reactions can be tolerated.
Humans ; Small Cell Lung Carcinoma/radiotherapy* ; Lung Neoplasms/radiotherapy* ; Quality of Life ; Treatment Outcome ; Progression-Free Survival

OBJECTIVE: To investigate the baseline levels of microorganisms' growth on the hands of anesthesiologists and in the anesthesia environment at a cancer hospital. METHODS: This study performed in nine operating rooms and among 25 anesthesiologists at a cancer hospital. Sampling of the hands of anesthesiologists and the anesthesia environment was performed at a ready-to-use operating room before patient contact began and after decontamination. RESULTS: Microorganisms' growth results showed that 20% (5/25) of anesthesiologists' hands carried microorganisms (> 10 CFU/cm ²) before patient contact began. Female anesthesiologists performed hand hygiene better than did their male counterparts, with fewer CFUs ( P = 0.0069) and fewer species ( P = 0.0202). Our study also found that 55.6% (5/9) of ready-to-use operating rooms carried microorganisms (> 5 CFU/cm ²). Microorganisms regrowth began quickly (1 hour) after disinfection, and increased gradually over time, reaching the threshold at 4 hours after disinfection. Staphylococcus aureus was isolated from the hands of 20% (5/25) of anesthesiologists and 33.3% (3/9) of operating rooms. CONCLUSION Our study indicates that male anesthesiologists need to pay more attention to the standard operating procedures and effect evaluation of hand hygiene, daily cleaning rate of the operating room may be insufficient, and we would suggest that there should be a repeat cleaning every four hours.
Female ; Humans ; Male ; Anesthesia ; Anesthesiologists/statistics & numerical data* ; Disinfection/standards* ; Hand Hygiene/statistics & numerical data* ; Staphylococcal Infections ; Operating Rooms/statistics & numerical data* ; Staphylococcus aureus/isolation & purification*

Objective:This work aims to assess the distribution of peripheral blood monocyte subsets, the expression level of the functional markers in rheumatoid arthritis (RA) patients, and analyze the correlation between the above indexes and the onset of RA.Methods:Peripheral blood mononuclear cells were collected and isolated from 62 RA patients, 52 healthy control (HC) and 12 disease control group′s patients via density centrifugation. The enrolled patients were attended or underwent physical examination in East Hospital, Tongji University from June 2020 to December 2021. Monocytes could be classified into classical (CM), intermediate (IM) and non-classical (NCM). Then, the flow cytometry was performed to examine the distribution of monocyte subsets and the measure the expression level of human leukocyte antigen DR (HLA-DR), intracellular tumor necrosis factor α (TNF-α) in peripheral blood monocytes. The statistical methods in this study mainly include: Kruskal-Wallis H test, Chi-Square test, Mann-Whitney U test, Wilcoxon matched-pairs signed ranks test, Spearman correlation coefficient test and Logistic regression analysis. The diagnostic value of IM proportion in RA was analyzed by ROC curve. Results:The monocytes number and monocytes proportion in white blood cells were much higher in RA [0.40 (0.40, 0.50), 7.60% (5.97%, 8.53%)] and disease control [0.40 (0.40, 0.68), 8.20% (5.85%, 10.28%)] compared with HC [0.30 (0.30, 0.40), 5.80% (5.03%, 6.38%)] ( H=24.733, P<0.001; H=27.469, P<0.001). A statistic-significant difference was detected among the proportion of CM[85.49%(76.91%,89.21%),88.94%(86.36%,91.72%),90.26%(80.25%, 92.56%)],IM[11.65%(8.47%,17.89%),7.89%(5.36%,10.75%), 5.56%(4.17%, 8.27%)], NCM[2.22%(1.39%, 3.74%), 2.49%(1.74%, 4.66%), 5.13%(3.39%, 9.85%)] in RA group, HC group and disease control group ( H=11.389, P=0.003; H=20.815, P<0.001; H=10.640, P=0.005). The proportion of CM was lower in RA and the IM proportion was increased in RA( P=0.003; P=0.003). The intracellular TNF-α level of monocytes in all three groups revealed the trend that IM>NCM>CM. The intracellular TNF-α in IM of RA was positively associated with serum TNF-α ( r=0.376, P=0.041). The HLA-DR expression in IM subsets were higher than CM and NCM subsets in all RA,HC and disease control groups. The expression of HLA-DR of IM in RA group and disease control was higher than HC group [8 611.50 (6201.3, 9890.8), 10 295.0 (7 899.0, 13632.0), 6 278.00(4 057.8, 9522.0), H=10.495, P=0.005]. There were no correlations between the proportion of peripheral blood IM and clinical characteristics CRP ( r=0.119, P=0.359), RF ( r=0.204, P=0.112) and ESR ( r=0.153, P=0.236). Logistic regression analysis showed that the proportion of IM ( OR=1.169, 95% CI 1.003-1.363, P=0.046), CRP ( OR=1.277, 95% CI 1.000-1.631, P=0.050), RF ( OR=1.179, 95% CI 1.080-1.287, P<0.001) are positively correlated with RA onset. The area under ROC curve for diagnosis of RA with IM proportion was 0.687, and the 95% confidence interval was 0.590-0.784, P<0.001. Conclusions:The distribution of monocyte subsets in peripheral blood of RA patients is abnormal. The increase in the proportion of IM, the enhanced antigen-presenting ability, and the increased level of TNF-α secretion in RA patients may play an important role in the pathogenesis of RA.

[Abstract] Objective: To analyze the expression level of miR-224 in cancer tissues and plasma of hepatocellular carcinoma (HCC) patients, and its correlation with clinicopathological characteristics, diagnosis and prognosis of HCC patients, and to further analyze its mechanism of action in the occurrence and development of liver cancer through bioinformatics analysis and in vitro experiments. Methods: The expression level of miR-224 in HCC tissues and normal tissues was analyzed using large sample data from Gene Expression Omnibus (GEO). qPCR method was used to verify the expression level of miR-224 in the tumor tissues and corresponding adjacent tissues that surgically resected from 80 HCC patients in Hebei Provincial People ’s Hospital from January 2017 to January 2020; in addition, the miR-224 level was also examined in plasma samples from 30 HCC patients. The Kaplan-Meier plotter database was used to analyze the correlation between the miR-224 expression and the overall survival time of HCC patients. The biological processes and signal pathways involving miR-224 were analyzed using bioinformatics tools. Hepatocellular carcinoma HepG2 cells were transfected with miR-224 inhibitor, and then Clone formation experiment, Transwell chamber experiment, qPCR and WB methods were used to detect the effect of miR-224 knockdown on the proliferation and invasion of HepG2 cells and the expression level of EMT-related molecules. Results: The results of GEO database analysis showed that the expression level of miR-224 in HCC tissues was significantly higher than that in normal tissues. The results of clinical specimen verification showed that the expression level of miR-224 in the tumor tissues and plasma of HCC patients was significantly higher than that in the corresponding adjacent tissues and plasma from healthy controls (all P<0.01). The expression level of miR-224 was significantly correlated with the TNM stage, lymph node metastasis status and tumor size of HCC patients (P<0.05 or P<0.01). ROC analysis indicated that miR-224 showed a prominent diagnostic value in liver cancer, and the increased expression level of miR-224 was significantly related to the poor prognosis of HCC patients (P<0.05). Functional enrichment analysis revealed that miR-224 was mainly involved in the mTOR signaling pathway, AGE-RAGE signaling pathway, Rap1 signaling pathway, Ras signaling pathway, ErbB signaling pathway, HIF-1 signaling pathway and p53 signaling pathway and other signaling pathways related to tumor occurrence and development. Knockdown of miR-224 could significantly inhibit the colony formation and invasion of HepG2 cells and affect the expression of EMT-related markers (P<0.05 or P<0.01). Conclusion: miR-224 is highly expressed in HCC tissues and plasma and is significantly related to the poor prognosis of HCC patients. Knockdown of miR-224 expression can inhibit the colony formation, invasion and EMT process of liver cancer HepG2 cells.

Objective:To explore the clinical significance of combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood for the diagnosis of colorectal cancer. Methods The data of patients admitted to the Department of Gastroenterology, Shanghai East Hospital Affiliated to Tongji University from January to September 2019 were retrospectively analyzed. They were divided into colorectal cancer group (62 cases of colon cancer, 59 cases of rectal cancer), precancerous lesions group (77 cases of colorectal adenoma, 5 cases of high-grade intraepithelial neoplasia), interference group (61 cases of colorectal cancer and advanced adenoma negative but suffered other intestinal lesions, 17 cases of non-colorectal cancer) and healthy group (94 cases). The methylation status of three genes (Septin9, SDC2 and BCAT1) in peripheral blood plasma was detected simultaneously by fluorescence PCR. The relationship between the positive rate of three genes detected jointly and the clinic pathological characteristics of colorectal cancer was analyzed and compared with serum carcinoembryonic antigen (CEA) positive rate. The colorectal cancer group was divided into stage Ⅰ, Ⅱ, Ⅲ and Ⅳ according to TNM stage, and the colorectal cancer group was analyzed and counted by grade. The diagnostic efficiency of detection methods was analyzed by receiver operating characteristic (ROC) curve, and the area under ROC curve (AUC) was compared.Results:The positive rate of combined detection of SDC2 and BCAT1 gene methylation was higher than other three groups (χ 2 =237.246, P<0.001). The positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation was higher than CEA in colorectal cancer group ( P<0.001). The positive rates of the combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅳ of colorectal cancer group were 73%(16/22), 87%(34/39), 86%(30/35) and 96%(24/25), respectively. Compared with CEA group, the positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅲ of colorectal cancer group was higher than serum CEA ( P<0.001), but the positive rate of stage Ⅳ was not statistically significant compared with CEA group ( P>0.05). ROC curve analysis showed that the AUC of Septin9, SDC2 and BCAT1 was 0.857(95% CI 0.810-0.903),0.819(95% CI 0.768-0.871)and 0.862(95% CI 0.816-0.909), respectively. The AUC of combined detection of three gene methylations was 0.889 (95% CI 0.846-0.933), and the AUC of combined detection with serum CEA was 0.913 (95% CI 0.874-0.951). There was no significant difference in the positive rate of combined detection of Septin9, SDC2 and BCAT1 gene methylation among different gender, age and cancerous site of colon cancer patients (all P>0.05). Conclusion:The combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood plasma is helpful for the early diagnosis of colorectal cancer. The positive rate in stage Ⅰ-Ⅲ of colorectal cancer group is higher than serum CEA. The combined diagnosis of the three genes can improve the diagnostic efficiency.

Objective To assess the current status and factors associated with the mental health condition of older adults. Methods A convenience sampling survey was conducted using symptom check list 90 (SCL-90) among the Chinese older adults aged 60 or above from January to March, 2018. The older adults aged 80 or above were selected for this study. Chi-square test and binary logistic regression model were used to analyze the influencing factors. Results The total participants were 485. The SCL-90 positive detection rate was 20.21%. The symptoms of the four highest detection rates were somatization (39.38%), others (25.15%), obsessive-compulsive symptoms (24.33%) and depression (22.68%). The older adults with normal BMI (OR=0.537, 95% CI: 0.250-0.857, P=0.027) and lived in county town (OR=0.224, 95% CI:0.075-0.667, P=0.007) showed lower SCL-90 positive detection rate. These who had been educated for 1-5 years (OR=11.092, 95% CI: 4.446-27.671, P<0.001), 6-8 years (OR=9.800, 95% CI: 3.464-27.721, P<0.001), 9~11 years (OR=19.279, 95% CI : 6.722-55.297, P<0.001), 12 years and above (OR=24.321, 95% CI: 7.894-74.929, P<0.001) had higher SCL-90 positive detection rate compared with those who were uneducated. Conclusion The mental health condition of Chinese older adults is mainly influenced by residence place, education level, family income self-evaluation and BMI status.

The combination of ginkgo ketoester tablet - donepezil (GD) is a popular combination commonly used in clinic for the treatment of Alzheimer's disease. To evaluate the learning and memory improving ability of different proportions of the two drugs. We optimized the ratio of GD for treatment of dementia using a mouse model. Dementia was induced by multiple neuronal damages in mice. The experimental protocols were approved by the Animal Experimental Ethical Committee of Nanjing University of Chinese Medicine and all the procedures were strictly conducted in accordance with ethical principle of animal use and care. Morris water maze, brain hematosylin-eosin staining and the changes of the neurotransmitters and related enzymes in the plasma or brain tissues were tested to determine the effect of GD on dementia mice. The results showed that the dementia mice were significantly different from the normal group in terms of behavior, pathological sections and related indicators. Compared to the dementia mice, partial administration groups could improve learning and memory ability as well as indexes in the blood and brain tissues. Both the principal component analysis and multi-attribute comprehensive index methods were used to comprehensively evaluate the total effect of GD on anti-dementia. The results showed that the combination of two drugs at the dose of 0.5 to 1 times was in a dose-effect relationship, and the dose of 1 (the clinical equivalent) had the best treatment effect. Then based on the optimal dose, GD 1∶1 had best effect, which was consistent with the clinical use of two drugs. This provides scientific basis for more effective application of the compatibility between ketoester tablet and donepezil for modern clinic medicine.