Objective:To analyze the etiological characteristics of Clostridium ramosum DZS3717106 isolated from the blood of a patient with septic shock secondary to perianal abscess and the immunological characteristics of the patient. Methods:The isolate was subjected to morphological observation, mass spectrometry, antibiotic susceptibility testing, and 16S rRNA sequencing. Biochemical and cytological test results of the patient were collected. Flow cytometry was used to detect T cell subsets. Impacts of the virulence factors of the isolate on the host immune system were evaluated.Results:DZS3717106 was an anaerobic Clostridium with pleomorphic rod-shaped cells and spores. It was sensitive to penicillin G, piperacillin/tazobactam, and metronidazole, but resistant to clindamycin. It carried various virulence and resistance genes. The patient was immunocompromised with abnormal IL-6, IL-8, and IL-10 levels. Conclusions:Septic shock caused by Clostridium ramosum is rare, and more research is needed on the causes and epidemiology. DZS3717106 infection triggers over-activated inflammatory response in the patient, which may be closely related to the occurrence and development of septic shock.

Objective:To analyze the etiological characteristics of Clostridium ramosum DZS3717106 isolated from the blood of a patient with septic shock secondary to perianal abscess and the immunological characteristics of the patient. Methods:The isolate was subjected to morphological observation, mass spectrometry, antibiotic susceptibility testing, and 16S rRNA sequencing. Biochemical and cytological test results of the patient were collected. Flow cytometry was used to detect T cell subsets. Impacts of the virulence factors of the isolate on the host immune system were evaluated.Results:DZS3717106 was an anaerobic Clostridium with pleomorphic rod-shaped cells and spores. It was sensitive to penicillin G, piperacillin/tazobactam, and metronidazole, but resistant to clindamycin. It carried various virulence and resistance genes. The patient was immunocompromised with abnormal IL-6, IL-8, and IL-10 levels. Conclusions:Septic shock caused by Clostridium ramosum is rare, and more research is needed on the causes and epidemiology. DZS3717106 infection triggers over-activated inflammatory response in the patient, which may be closely related to the occurrence and development of septic shock.

Objective:To investigate the factors that influence the prolongation of gestation following emergency cervical cerclage.Methods:This retrospective study included 88 singleton pregnant women who were diagnosed with cervical incompetence and underwent emergency cervical cerclage at 12-26 weeks of gestation in the Jiaxing Maternity and Child Health Care Hospital from January 1, 2019 to September 1, 2022. The participants who delivered after 28 gestational weeks were assigned to the success group ( n=77), while those who delivered or miscarried before 28 gestational weeks were assigned to the failure group ( n=11). Two independent sample t-test or Mann-Whitney U test was used for comparison between groups. The factors affecting the prolongation of gestation after the procedure were selected by univariate analysis and multiple regression equations. Results:The success rate (delivery rate≥28 weeks) was 87.5% (77/88). There were six women delivered at 28-31 +6 weeks of gestation (6/77, 7.8%), two at 32-33 +6 weeks of gestation (2/77, 2.6%), 16 at 34-36 +6 weeks of gestation (16/77, 20.8%), and 53 at 37 weeks of gestation or later (53/77, 68.8%). Multiple regression analysis indicated that three times of early pregnancy miscarriage ( β=－5.1, 95% CI: -9.5 to -0.7), five times of early pregnancy miscarriage ( β=－11.8, 95% CI: -22.1 to -1.6), had two live births ( β=－6.9, 95% CI: -12.9 to -0.9), gestational age at cerclage ( β=－0.6, 95% CI: -1.0 to -0.3), external cervical os dilation<15 mm ( β=－12.1, 95% CI: -22.5 to -1.8) and ≥15 mm before cerclage ( β=－11.0, 95% CI: -21.4 to -0.71) were factors affecting the prolongation of gestation after cerclage. After adjusting for maternal age and weight before emergency cervical cerclage, five times of early pregnancy miscarriage ( β=－18.1, 95% CI: -28.3 to -7.8), gestational age at cerclage ( β=－0.6, 95% CI: -1.0 to -0.3), and external cervical os dilation≥15 mm before cerclage ( β=－11.4, 95% CI:－21.2 to -1.6) remained the significant influencing factors (all P<0.05). Conclusion:The number of early pregnancy miscarriages, gestational age at cerclage, and the width of external cervical os dilation before cerclage≥15 mm are the factors that influence the prolongation of gestation after the emergency cervical cerclage procedure.

Objective:To evaluate the expressions of three biomarkers combination of CD27, CD38 and human leucocyte antigen (HLA)-DR in the application of discrminating active tuberculosis (ATB) and latent tuberculosis infection (LTBI).Methods:Sixty cases of ATB and 44 cases of LTBI were enrolled from March 2021 to February 2022 in Huashan Hospital, Fudan University and Wuxi Fifth People′s Hospital. Freshly isolated peripheral blood mononuclear cells (PBMC) from patients were stimulated with 6 kDa early secretory antigenic target/culture filtrate protein 10 peptide pools. The expressions of CD27, CD38 and HLA-DR on Mycobacterium tuberculosis-specific CD4 + T lymphocytes were evaluated by polychromatic flow cytometry. Mann-Whitney U test was used for statistical analysis. The area under the receiver operator characteristic curve (AUROC) was used to evaluate the diagnostic value of biomarkers in discriminating ATB and LTBI. Results:The frequencies of CD27 -, CD38 +, HLA-DR +, CD27 -CD38 +, CD27 -HLA-DR + and CD38 + HLA-DR + in ATB group were all higher than those in LTBI group, and the differences were all statistically significant ( U=26.00, 451.00, 384.00, 8.00, 7.00 and 184.00, respectively, all P<0.001). The AUROC of CD27 -CD4 + interferon-γ(IFN-γ) + T lymphocytes was 0.71 with a cut-off value of 52.31%, with the sensitivity of 50.00% and specificity of 87.20%. The AUROC of CD38 + CD4 + IFN-γ + T lymphocytes was 0.82 with a cut-off value of 30.25%, with the sensitivity of 73.40% and specificity of 89.70%. The AUROC of HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.85 with a cut-off value of 36.60%, with the sensitivity of 66.00% and specificity of 94.90%. The AUROC of CD27 -CD38 + CD4 + IFN-γ + T lymphocytes was 0.80 with a cut-off value of 8.82%, with the sensitivity of 90.60% and specificity of 61.50%. The AUROC of CD27 -HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.83 with a cut-off value of 18.62%, with the sensitivity of 75.00% and specificity of 79.50%. The AUROC of CD38 + HLA-DR + CD4 + IFN-γ + T lymphocytes was 0.93 with a cut-off value of 22.35%, with the sensitivity of 79.70% and specificity of 100.00%. Conclusions:The expressions of CD27 -, CD38 + and HLA-DR + in Mycobacterium tuberculosis-specific CD4 + T lymphocytes are higher in ATB group compared to LTBI group. ATB and LTBI could be well discriminated by detecting the expressions of CD27, CD38 and HLA-DR on CD4 + IFN-γ + T lymphocytes with flow cytometry.

Objective:To investigate the role of glycoprotein A repetitions predominant (GARP) in the pathogenesis of tuberculosis through regulatory T cell (Treg), in order to provide new targets for the treatment of tuberculosis.Methods:Sixty patients with active pulmonary tuberculosis (ATB) admitted to Huashan Hospital, Fudan University and Wuxi Fifth People′s Hospital from January to September 2021 were included. And six individuals with latent tuberculosis infection (LTBI), and 16 healthy controls (HC) were recruited during the same period. Flow cytometry was performed to detect the proportion of Treg in the peripheral blood, and the expressions of GARP and transforming growth factor-β1 (TGF-β1) on Treg in different groups. Mann-Whitney U test was used for statistical analysis. Results:Among the 60 patients with ATB, 23 patients did not receive anti-tuberculosis drug therapy, 17 patients were treated for less than three months, ten patients were treated for three to less than six months, and ten patients were treated for greater than or equal to six months. The percentage of CD4 + CD25 + forkhead box protein 3 (Foxp3) + Treg in untreated ATB patients was 7.50%(5.67%, 9.00%), which was higher than that in HC (5.57%(5.03%, 6.09%)), and the difference was statistically significant ( U=95.00, P=0.010). The percentage of GARP expressing in CD4 + CD25 + Foxp3 + Treg in untreated ATB patients was 10.37%(7.79%, 12.90%), which was higher than that in LTBI (7.02%(5.15%, 8.81%)) and HC (5.33%(4.26%, 6.67%)), respectively, and the differences were both statistically significant ( U=31.00, P=0.040; U=36.00, P<0.001, respectively), while there was no significant difference between LTBI and HC ( U=25.00, P=0.095). The percentage of CD4 + CD25 + Foxp3 + Treg expressing TGF-β1 in untreated ATB patients was 7.13%(4.25%, 8.89%), which was higher than that in HC (3.59%(2.10%, 5.17%)), and the difference was statistically significant ( U=71.00, P=0.001). The expressions of GARP in CD4 + CD8 -CD25 + Foxp3 + Treg in patients with ATB treated for less than three months group, three to less than six months group and greater than or equal to six months group were 7.82%(3.94%, 13.17%), 6.92%(5.61%, 9.47%) and 7.26%(5.82%, 9.64%), respectively. The expressions of TGF-β1 in CD4 + CD8 -CD25 + Foxp3 + Treg in the above three treatment groups were 11.16%(7.91%, 15.23%), 8.66%(5.43%, 12.54%) and 7.82%(6.01%, 9.53%), respectively, and the expression of TGF-β1 in CD4 + CD8 -CD25 + Foxp3 + Treg in the patients with ATB treated for less than three months group was higher than that in the greater than or equal to six months group, the difference was statistically significant ( U=37.50, P=0.024). Conclusions:Foxp3/GARP/TGF-β1 pathway may be involved in the immune mechanism of Treg regulating the pathogenesis of tuberculosis, and GARP may be a new target for anti-tuberculosis therapy.

Objective:To investigate the diagnostic values of interleukin-22 (IL-22), interferon-γ(IFN-γ)and macrophage migration inhibition factor (MIF) in pleural effusion for tuberculosis pleurisy.Methods:From April 2018 to May 2019, a total of 77 patients including 45 cases of tuberculous pleurisy, 19 cases of malignant pleurisy, 13 cases of parapneumonia and 13 cases of healthy control in Wuxi Fifth People′s Hospital were enrolled. The levels of IL-22, IFN-γ and MIF in plasma and pleural effusion were detected by enzyme linked immunosorbent assay (ELISA). Mann-Whitney U test was used for statistical analysis.The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic values of IL-22, IFN-γ and MIF for tuberculous pleurisy. Results:The median levels of IL-22, IFN-γ, MIF and adenosine deaminase in 45 cases with pleural effusion in tuberculosis pleurisy group were 396.8 ng/L, 2 200.0 ng/L, 241.3 μg/L and 70.8 U/L, respectively, which were all significantly higher than 32 cases with non-tuberculosis pleurisy group, including 19 cases with malignant pleurisy and 13 cases with parapneumonia (52.8 ng/L, 232.3 ng/L, 179.6 μg/L and 17.0 U/L, respectively). The differences were all statistically significant ( U=179.000, 118.500, 287.000, 162.000, respectively, all P<0.05). The median levels of IL-22 and IFN-γ in plasma of tuberculosis pleurisy group were 20.0 ng/L and 45.9 ng/L, respectively, which were both higher than healthy control group (14.3 ng/L and 33.4 ng/L, respectively). The level of MIF was 96.2 μg/L, which was lower than healthy control (159.5 μg/L). The differences were all statistically significant ( U=74.000, 13.000 and 73.000, respectively, all P<0.05). The areas under ROC curve (AUC) of IL-22, IFN-γ and MIF in pleural effusion for the diagnosis of tuberculosis pleurisy were 0.876, 0.917 and 0.682, respectively.The sensitivities were 93.75%, 100.00% and 63.64%, respectively; the specificities were 82.22%, 91.11% and 65.85%, respectively. The median levels of IL-22 and IFN-γ in plasma in tuberculosis pleurisy group at two months of follow-up after anti-tuberculosis therapy were 16.0 ng/L and 33.9 ng/L, respectively, which were both lower than baseline (20.0 ng/L and 44.7 ng/L, respectively). The differences were both statistically significant ( U=2.156 and 2.221, respectively, both P<0.05). Conclusion:IFN-γ and IL-22 in pleural effusion could be used as effective indicators to identify tuberculous pleurisy, and the dynamic monitoring of IL-22 in patients′plasma could be an important biomarker in evaluating the efficacy of anti-tuberculosis treatment.

Objective To explore the effect of sequential and comprehensive preventative measures on the development of premature infants' intelligence.Methods A cohort of 120 premature infants was randomly divided into an observation group and a control group,each of 60.Both groups were given routine premature infant care,but the observation group was additionally provided with sequential and comprehensive preventive intervention.It included neonatal screening,inpatient-outpatient link-up,and their parents' watching CDs explaining early childhood education and health education.All of the infants were followed up from birth to 3 years old.Their adaptive capacity,fine motor skills,language acquisition,gross motor skills and social communication were evaluated at 12,24 and 36 months old using a child intelligence developmental scale for neurological development.Development intelligence quotients (DQs) were calculated and compared.Results After 12 months,significant inter-group differences were observed in adaptability and fine motor control.At 24 and 36 months old there were also significant differences in language skills.At one,two and 3 years old the average DQ of the observation group was significantly higher than that of the control group.Significant within-group differences in average DQ were observed in both groups between 1 and 2 years old,but not between 2 and 3.Conclusion Intervention within two years after birth is critical for premature infants.Timely,sequential,integrated,preventive intervention can promote the development of intelligence and better life quality for premature infants.

Objective To analyze the expressions of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) in the peripheral blood of patients with active tuberculosis (ATB ) or latent tuberculosis infection (LTBI) ,and to evaluate its diagnostic value in differentiation of ATB and LTBI .Methods Forty-eight patients including 18 ATB cases and 30 LTBI cases were continuously enrolled from Wuxi No . 5 People′s Hospital and Huashan Hospital affiliated to Fudan University from January 2011 to March 2013 .Flow cytometry was applied to detect the CTLA-4 expression in CD4+CD25+ FoxP3+ T cells in the peripheral blood of the 48 subjects .CTLA-4 levels were compared using non-parametric Mann-Whitney U test .Results The median percentage of CTLA-4+ Treg in CD4+ CD25+ Foxp3+ Treg cells of ATB patients was 18 .95％ (quantile range :13 .86％ ,27 .73％ ) ,and that in LTBI patients was 6 .67％(quantile range :5 .74％ ,9 .59％ ) ,which was statistically significant (U=18 .0 , P< 0 .01) .Receiver operating curve (ROC) based on the CTLA-4 expression indicated that the area under the curve was 0 .96 , with the optimum cut-off value of 13 .25％ .Thus ,the sensitivity and specificity for the diagnosis of ATB were 86 .7％ and 94 .4％ ,respectively .Conclusion CTLA-4 has highly sensitivity and specificity for the differential diagnosis of ATB and LTBI whose interferon-gamma releasing assays are all positive ,which may also provide meaningful clue for the study of pathogenesis of ATB .

Thirty eight patients with type 2 diabetes mellitus (T2DM) signed with the Lingering Garden Subdistrict Community Health Service Center of Suzhou Gusu District from October to December 2015 were enrolled in the study. The patients were classified as low risk group (n=3), moderate risk group (n=11), high risk group (n=21) and extremely high risk group (n=3) according to risk stratification of the JADE program based on estimated glomerular filtration rate (eGFR) and risk factors [obesity, dyslipidemia, hypertension, diabetic retinopathy, urinary protein creatinine ratio, foot disease, glycosylated hemoglobin A1c(HbA1c), fasting plasma glucose(FPG) or non fasting plasma glucose]. The patients were managed by a team consisting of general practitioners, general nurses and specialists; the individualized management was implemented with reduction of controllable risk factors and complications as the goal. After one year of management (March 2016 to February 2017) the indicators of T2DM were evaluated. The results showed that the TC, LDL-C, FPG and HbA1c levels were significantly improved in both moderate and high risk groups (P<0.05); the TC, LDL-C and FPG levels in the extremely high risk group were also significantly improved (P<0.05).

Objective To explore the role of basic leucine zipper ATF-like transcription factor (BATF) in active tuberculosis, and to provide clues for diagnosis and therapy of tuberculosis.Methods Sixteen patients with active tuberculosis (ATB), ten cases of latent tuberculosis infection (LTBI) and fourteen healthy controls (HC) were included in this study.Flow cytometry was applied to detect the expressions of BATF and programmed death-1 (PD-1) in T lymphocytes, and the changes of BATF by blockade of PD-1/PD-L pathway using specific blocking antibody antiPD-1, antiPD-L1 and antiPD-L2.The expressions of BATF were compared using Mann-Whitney U test.And the relation of BATF and PD-1 was analyzed using Pearson correlation analysis.Results The CD4+ T lymphocytes expressing BATF accounted for 5.16% (2.96%,8.71%) of CD4+ T lymphocytes in ATB group, which was higher than 1.05% (0.40%,1.27%) in LTBI group and 0.71%(0.43%,1.21%) in HC group, and the difference were statistically significant (U value were 6.5 and 9.0, respectively, both P<0.01).The CD8+ T lymphocytes expressing BATF accounted for 4.10% (2.27%,8.17%) of CD8+ T lymphocytes in ATB group, which was higher than 0.55% (0.34%,1.18%) in LTBI group and 0.84% (0.41%,1.29%) in HC group, and the difference were statistically significant (U value were 5.0 and 8.0, respectively, both P<0.01).Furthermore, the percentage of BATF+ PD-1+ CD4+ T lymphocytes in the peripheral blood of ATB was significantly higher than those in the peripheral blood of LTBI and HC, the difference were statistically significant (Uvalue were 16.0 and 14.5, respectively, both P<0.01), and the percentage of BATF+ PD-1+ CD8+ T lymphocytes in the peripheral blood of ATB was significantly higher than those in the peripheral blood of LTBI and HC, the difference were statistically significant (Uvalue were 10.0 and 16.5, respectively, both P<0.01).In addition, there was a positive correlation between the percentage of BATF+ T cells and PD-1+ T cells, both in CD4+ T cells (r=0.676,P=0.016) and CD8+ T cells (r=0.610,P=0.035).The expressions of BATF both in CD4+ T cells and CD8+ T cells were decreased followed by blockade of PD-1/PD-L pathway (P<0.05).Conclusions BATF may be involved in the regulation of immune pathogenesis of tuberculosis.In order to provide a theory for anti-tuberculosis immunotherapy fargeting BATF, further research need to be proceeded.