Objective To establish a real‐time fluorescent polymerase chain reaction and detect 16S rRNA gene of Legionella strains isolated from sputum specimens of patients with pulmonary infection by using this method .Methods 16s rRNA gene of Le‐gionella was used to design primers and probes .The reaction system and reaction conditions were optimized and the specificity ,sen‐sitivity and repeatability of this method were verified by detecting Legionella pneumophila ,non‐Legionella pneumophila and other bacteria .A total of 557 sputum specimens of patients with pulmonary infection were detected ,and PCR‐digestion identification method was carried out as control .Otherwise ,sequences of 16S rRNA were verified in patients with positive detection results .Re‐sults The results showed that all reference strains of Legionella were positive ,while all of other bacteria were negative ,and the sensitivity was 102 CFU/mL .Among sputum specimens collected from 577 cases of patients with pulmonary infection ,the positive rate of Legionella detected by using real‐time fluorescent PCR and PCR‐digestion identification method was 23 .1% and 19 .9% re‐spectively ,while the positive rate was 17 .2% by verifying the sequences of 16s rRNA .There were no statistically significant differ‐ences of positive rate among the three methods(P>0 .05) .Conclusion The real‐time fluorescent PCR is fast and convenient in de‐tection of Legionella strains isolated from sputum specimens of patients ,which could be an assisted method for clinically diagnosing Legionella infection .

BACKGROUND: Although we have gained much information about leadinduced organ damage, the effect of blood lead level on T cell subgroup is yet to be determined.OBJECTIVE: To assess the effect of blood lead level on T cell subgroup of children, and the association of T cell subgroup with threshold limit value of blood lead level.DESIGN: Comparative observation.SETTING: The Institute of Molecular Microorganism, Maternity and Child Care Hospital, Shanxi Provincial Children Hospital.PARTICIPANTS: Sixty children, (32 boys and 28 girls), aged 3-6 years with a mean of (4.5±1.5) years old, were selected from the suburbs of Taiyuan city between May 2003 and October 2003. Informed consents were obtained from all their guardians. The enrolled children according to their blood lead levels were assigned into three groups, 13 in Group Ⅰ with blood lead level ≥ 0.48 μmol/L, 20 in Group 2 with lead level ≥ 0.24 μmol/L but ＜ 0.48 μmoL/L and 27 in Group 3 with lead level ＜ 0.24 μmol/L.METHODS: Blood lead level and expression of T cell subgroup were measured with graphite furnace atomic absorption spectroscopy and immunofluorescence methods respectively. Student t test was used in data analysis, and linear correlation analysis was used to assess the correlation between blood lead level and expression of T cell subgroup.level and expression of T cell subgroup.pression of CD3 and ratio of CD4 to CD8 were lower in Group 1 (lead level ≥0.48 μmol/L) than Group 3 (lead level ＜ 0.24 μmol/L) (t=3.27,blood lead level showed had significant inverse correlation with CD3 expression and the ratio of CD4 to CD8(r=-0.689,-0.674,P ＜ 0.01).CONCLUSION: A blood lead level ≥ 0.48 μmol/L, is shown to significantly decrease the expression of CD3 and ratio of CD4 to CD8 in peripheral blood, which may impair the children's immunological function.

BACKGROUND: Although we have gained much information about leadinduced organ damage, the effect of blood lead level on T cell subgroup is yet to be determined.OBJECTIVE: To assess the effect of blood lead level on T cell subgroup of children, and the association of T cell subgroup with threshold limit value of blood lead level.DESIGN: Comparative observation.SETTING: The Institute of Molecular Microorganism, Maternity and Child Care Hospital, Shanxi Provincial Children Hospital.PARTICIPANTS: Sixty children, (32 boys and 28 girls), aged 3-6 years with a mean of (4.5±1.5) years old, were selected from the suburbs of Taiyuan city between May 2003 and October 2003. Informed consents were obtained from all their guardians. The enrolled children according to their blood lead levels were assigned into three groups, 13 in Group Ⅰ with blood lead level ≥ 0.48 μmol/L, 20 in Group 2 with lead level ≥ 0.24 μmol/L but ＜ 0.48 μmoL/L and 27 in Group 3 with lead level ＜ 0.24 μmol/L.METHODS: Blood lead level and expression of T cell subgroup were measured with graphite furnace atomic absorption spectroscopy and immunofluorescence methods respectively. Student t test was used in data analysis, and linear correlation analysis was used to assess the correlation between blood lead level and expression of T cell subgroup.level and expression of T cell subgroup.pression of CD3 and ratio of CD4 to CD8 were lower in Group 1 (lead level ≥0.48 μmol/L) than Group 3 (lead level ＜ 0.24 μmol/L) (t=3.27,blood lead level showed had significant inverse correlation with CD3 expression and the ratio of CD4 to CD8(r=-0.689,-0.674,P ＜ 0.01).CONCLUSION: A blood lead level ≥ 0.48 μmol/L, is shown to significantly decrease the expression of CD3 and ratio of CD4 to CD8 in peripheral blood, which may impair the children's immunological function.

Objective To inves1tigate the needs and influencing factors of patients with cerebrovascular disease and their caregivers in rural areas.Methods An online questionnaire designed by World Stroke Organization for stroke patients and their caregivers was used,and modified appropriately according to China's national conditions.A questionnaire survey about general situation,and needs for psychological support,stroke-related knowledge,diagnosis and nursing,social support,and functional recovery in inpatients with cerebrovascular disease and their caregivers in rural areas was performed.Results A total of 514 patients with stroke and their caregivers (n =514) completed the questionnaires.93.0％ of the patients had the needs for mental support,followed by clinical diagnosis and nursing (84.8％),stroke knowledge (74.7％),social support and functional recovery (53.5％);95.1％ of the caregivers had the needs for mental support,followed by stroke knowledge (89.9％),clinical diagnosis and nursing (84.0％),social support,and functional recovery (66.3％).The degree of need for stroke knowledge,social support,and functional recovery in caregivers was higher than that in the patients (all P ＜ 0.05).Multiple linear regression analysis showed that age,gender,degree of education,the National Institutes of Health Stroke Scale score,and the time from the first stroke onset were the influencing factors of needs for patients with cerebrovascular disease and their caregiver.Conclusions Many needs have not been met in patients with cerebrovascular disease and their caregivers,especially the psychological demand.The regional and individual services should be provided according to the different demands in patients with cerebrovascular disease and their caregivers,enhance the awareness of the prevention and treatment of stroke,and continuously improve the construction of rural health service system.

Objective:To investigate the predictive value of regional leptomeningeal collateral (rLMC) score on CT angiography (CTA) for the outcome after endovascular therapy in patients with late window anterior circulation stroke.Methods:Patients with acute anterior circulation large vessel occlusive stroke received endovascular treatment 6 to 24 h after onset in the Department of Neurology, the First Affiliated Hospital of Soochow University from January 2018 to July 2021 were enrolled retrospectively. At 3 months after onset, the outcome was evaluated according to the modified Rankin Scale. A score of ≤ 2 was defined as good outcome, and a score of >2 was defined as poor outcome. The clinical data, non-enhanced CT and CTA parameters of the patients were collected. Multivariate logistic analysis was used to determine the independent influencing factors of poor outcomes after endovascular treatment. Results:A total of 74 patients with acute anterior circulation large vessel occlusive stroke treated with endovascular treatment in the late window were enrolled. Their age was 64.41±12.98 years, 43 were males (58.1%). The baseline National Institutes of Health Stroke Scale (NIHSS) score was 13.18±5.22, and the median time from onset to puncture was 527.5 min. Fifty-three patients (71.6%) chose direct thrombectomy and 21 (28.4%) chose intravenous thrombolysis and bridging thrombectomy. Thirty-six patients (48.6%) had a good outcome and 38 (51.4%) had a poor outcome, including 4 (5.4%) died. Univariate analysis showed that there were significant differences in age, atrial fibrillation, fasting blood glucose, NIHSS score, Alberta Stroke Program Early CT Score (ASPECTS), rLMC score and clot burden score between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that higher ASPECTS (odds ratio 0.352, 95% confidence interval 0.157-0.791; P=0.012) and rLMC score (odds ratio 0.550, 95% confidence interval 0.329-0.919; P=0.022) were the independent predictors of good outcomes after endovascular therapy. Conclusion:ASPECTS and rLMC scores were the independent predictors of clinical outcomes after endovascular therapy in patients with late window anterior circulation large vessel occlusive stroke. It had certain guiding value for the decision-making of endovascular treatment in such patients.

Objective:To investigate the expression of costimulatory molecule OX40 in peripheral CD4 +CD25 +CD127 low regulatory T (Treg) cells and its clinical significance in patients with acute cerebral infarction (ACI). Methods:Seventy-five patients with first-onset ACI, admitted to our hospital from April 2019 to December 2019, and 36 age- and gender-matched volunteers (control group) were selected in this study. OX40 expression on CD4 +CD25 +CD127 low Treg cells in peripheral blood samples in the two groups were analyzed by immunofluorescent labeling and flow cytometry. Correlations of OX40 +Treg cell percentage with National Institute of Health stroke scale (NIHSS) scores, ischemic penumbra volume, core infarct volume, and infarct volume in the patient group were analyzed. The changes of OX40 +Treg cell percentage in the patient group before and after endovascular treatment or intravenous thrombolysis were compared. Results:As compared with that in the control group, the Treg cell percentage in peripheral blood samples of the patient group was significantly decreased, while OX40 +Treg cell percentage was significantly increased ( P<0.05). OX40 +Treg cell percentage was positively correlated with NIHSS scores in ACI patients ( r s=0.271, P=0.018). Meanwhile, OX40 +Treg percentage was significantly correlated with ischemic penumbra volume, core infarct volume, and infarct volume in the patient group ( r s=0.435, P=0.000; r s=0.343, P=0.003; r s=0.245, P=0.034). OX40 +Treg cell percentage in ACI patients 7 d after endovascular treatment was significantly decreased as compared with that before treatment ( P<0.05); OX40 +Treg percentage in ACI patients 3 and 7 d after intravenous thrombolysis was significantly decreased as compared with that before treatment ( P<0.05). Conclusion:OX40 is abnormally expressed on peripheral Treg cells in ACI patients, and is closely correlated with neurological deficits, imaging features and reperfusion therapy.