Objective To provide anatomic data for operation of inserting the electron cochlear in young children. Methods Fourteen heads,28 sides specimens of young children of 1to-5-year old were dissected,through posterior tympanum approach,via mastoidectomy,posterior tympanoto to enter posterior tympanum.The related anatomy structures of the location of the electron cochlear inserted into the proper sites were observed and measured under surgical microscope. Results The round window was seated in superior part of the round window niche.The pyramidal eminence,tendo musculi stapedius,incudostapedial joint,base of stapes,cochleariform process,round window niche and promontorium tympani were all visible from different directions.The posterior arch of stapes was situated in the prozone of scala.Scala was situated in the posteroinferior scala vestibuli.The distance from the middle point of the anterior border of the round window niche to the inferior wall was(1.49?0.42)mm,to the posterior wall of the Scala tympani(0.90?0.31)mm,to the basal tissue(1.49?0.41)mm,to the pyramidal eminence(3.28?0.55)mm,to the lateral semicircular canal(7.41?0.90)mm,to the inferior margin of the base of stapes(3.09?0.53)mm.Conclusion It is considered that the location of the insertion should be at the middle point of anterior border of the round window niche anterior from 0.90mm to 1.49mm,deorsum from 0mm to 1.49mm.When the round window niche is not found,the location of the insertion has to be at the middle point of the inferior margin of the base of stapes deorsum 3mm.

Objective To investigate the influence of the purging fire and removing toxin method on chemokines and adhesion factors related to vascular endothelialitis injury induced by toxin of trimeresurus stejnegeri bite.Methods ① Animal experiment:50 healthy New Zealand white rabbits were chosen.According to random numbers generated by statistical software,they were divided into normal control group,model group,low,middle and high dose Sheshang capsule groups,10 in each group.Trimeresurus stejnegeri bite model was replicated by injecting 0.75 mL/kg snake venom into subcutaneous tissues of rabbits' right hind legs.And the same volume of normal saline was injected into the rabbit in the normal control group.After the model was established for 6 hours,the rabbits in low,middle and high dose Sheshang capsule groups received 174,348 and 522 mg· kg-1 · d-1 of Sheshang capsule solution respectively (the content of capsules was dissolved in normal saline to make liquid with 17.4,34.8 and 52.2 g/L Sheshang solution respectively,so the volume of gavage of each group was 10 mL· kg-1 · d-1);in the model and normal control groups,the same amount of normal saline was given by gavage,once daily for consecutive one week.24 hours after the last gavage,the blood of the rabbits was collected through an auricular border vein and the serum was separated by centrifuge ready for use.Meanwhile,the whole abdominal aorta segment of the rabbit was harvested and kept them in liquid nitrogen ready for use.② Cell experiment:human umbilical vascular endothelial cell (HUVEC) was cultured with MEM for 24 hours.The solution was replaced and according to the random number generated by statistical software,the cells were divided into blank control group,model group and low,middle,high dose Sheshang capsule medicinal serum groups,10 wells in each group.Trimeresurus stejnegeri toxin cell model was reproduced by addition of 5 mg/L snake venom into the cell culture medium.After 6-hour culture,the cells of model group and blank control group received 10％ normal rabbit serum,and the cells of low,middle and high dose Sheshang medicinal serum capsule groups received serum containing 5％,10％ and 15％ drug,respectively.After culture for 72 hours,the cells were collected and the total RNA was extracted.The real-time fluorescent quantitative polymerase chain reaction (qPCR) was used to detect the levels of mRNA of interleukin-8 (IL-8),monocyte chemoattractant protein-1 (MCP-1),intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial cell adhesion molecule-1 (VCAM-1) in the vascular endothelial cells of rabbit aorta abdominalis and human umbilical vein,and the content of serum E-select element (CD62E) was measured by enzyme linked immunosorbent assay (ELISA).Results In model group,the expression levels of mRNA in IL-8,MCP-1,ICAM-1,VCAM-1 and the content of CD62E were all increased significantly in the endothelial cells of rabbit aorta abdominalis and HUVEC compared with those in control group [when the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 in normal and blank control group were all being 1,the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in animal model group were 3.96 ± 0.39,3.07 ± 0.27,3.71 ± 0.26,3.94 ± 0.26,and the mRNA expression levels (2-△ △Ct) of the above mentioned inflammatory factors and adhesion molecule in HUVEC model group were 3.53±0.70,2.24±0.48,3.13±0.44,2.80±0.13,respectively,all P ＜ 0.01].The content of CD62E in serum was increased significantly in model group compared with that in normal control group (μg/L:1.31 ± 0.22 vs.0.82 ± 0.13,P ＜ 0.01),the mRNA expression levels of IL-8,MCP-1,ICAM-1 and VCAM-1 were decreased significantly in low,middle,high dose Sheshang capsule groups compared with those in model group in endothelial cells of aorta abdominalis of rabbits and HUVEC [abdominal aorta:IL-8 mRNA (2-△ △Ct) were 1.13 ± 0.19,1.26 ± 0.16,1.27 ± 0.17 vs.3.96 ± 0.39,MCP-1 mRNA (2-△ △ Ct) were 1.79 ± 0.24,2.22 ± 0.38,1.76±0.19 vs.3.07±0.27,ICAM-1 mRNA (2 △△Ct) were 2.05±0.11,1.68±0.09,2.37±0.48 vs.3.71±0.26,VCAM-1 mRNA (2-△△Ct) were 1.59±0.08,1.40±0.11,1.84±0.11 vs.3.94±0.26;HUVEC:IL-8 mRNA (2-△△Ct) were 2.33±0.59,2.82±0.82,2.51±0.77 vs.3.53±0.70,MCP-1 mRNA (2-△△Ct) were 1.59±0.35,1.48±0.36,1.54±0.29 vs.2.24±0.48,ICAM-1 mRNA (2-△△Ct) were 1.46±0.38,1.77±0.65,1.73±0.50 vs.3.13±0.44,VCAM-1 mRNA (2-△△Ct) were 2.49±0.24,2.18±0.19,2.45±0.24 vs.2.80±0.13,all P ＜ 0.05].The contents of CD62E were decreased significantly in middle,high dose Sheshang capsule groups compared with the content in model group (μg/L:1.01 ±0.14,1.04±0.13 vs.1.31 ±0.22,all P ＜ 0.01),but there were no statistical significant differences among the three drug group (all P ＞ 0.05).Conclusion The therapy of purging fire and removing toxin can treat vascular endothelial injury by inhibiting the inflammatory response induced by Trimeresurus stejnegeri bites.

ObjectiveTo evaluate the application of a whole blood interferon-γ (IFN-γ) release assay QuantiFERON-TB Gold In Tube (QFT-GIT) in the diagnosis of tuberculous pleural effusion.Methods IFN-γ released by specific T cells stimulated by early secreted antigenic target 6 × 103protein (ESAT-6),culture filtrate protein 10 (CFP -10) and TB7.7 were measured by QFT-GIT test in 44 tuberculous pleural effusion patients and 16 non-tuberculous pleurisy controls.The IFN-γ release level between groups was compared by Mann-Whitmey test.ResultsThe positive rates of QFT-GIT in patients with tuberculous pleural effusion and non tuberculous pleurisy were 95.5％ and 12.5％,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of QFT-GIT were 95.6％,87.5％,95.6％ and 87.5％,respectively.The antigen-specific IFN-γ release level in the patients with tuberculous pleural effusion was significantly higher than that in non-tuberculous pleurisy controls (P＜0.01).Conclusions The whole blood INF-γ release assay QFT-GIT is a sensitive and specific assay for detecting pleural tuberculosis infection.It could be a useful diagnostic tool for the diagnosis of tuberculous pleural effusion in China.

Objective To investigate the mechanism of traditional Chinese medicine (TCM) Sheshang capsule for treatment of blood coagulation dysfunction in patients bitten by Trimeresurus stejnegeri snake. Methods A prospective study was conducted. Seventy Trimeresurus stejnegeri snake envenoming patients whose manifestations conformed to the diagnostic criteria of the fire toxin syndrome in TCM were assigned into therapy group and control group by random number table (each, 35 cases). The basic treatments (including wound disinfection, intramuscular injection of 1 500 U tetanus antitoxin, conventional dose of antibiotics, 10 mg dexamethasone, 40 mg omeprazole) and 10 Jidesheng Sheyao tablets three times a day were applied in the control group. In the therapy group, the basic treatments the same as those of the control group were given, and in the mean time 5 Sheshang capsules (the drug was prepared in our hospital including ingredients:rhubarb, coptidis rhizoma, pleione bulbocodioides, elecampane inula root, bayberry bark, borneol and so on) were administered three times a day. The therapeutic course in the two groups was 1 week. The levels of platelet α-granule membrane protein (CD62p), thromboxane B2 (TXB2), platelet factor 3 (PF3) and von Willebrand factor (vWF) in serum were measured by enzyme linked immunosorbent assay (ELISA) before and after treatment. Results Before treatment, there were no significant differences in CD62p, TXB2, PF3 and vWF between therapy group and control group [CD62p (μg/L):3.81±1.64 vs. 3.52±1.43, TXB2 (μg/L):13.04±1.67 vs. 13.31±1.14, PF3 (μg/L): 2.84±1.08 vs. 2.88±1.23, vWF (μg/L):12.36±2.42 vs. 11.89±2.08, all P>0.05]. After treatment, the levels of CD62p, TXB2 and PF3 were increased, while vWF decreased compared with those before treatment in both groups, the level changes in therapy group being more remarkable [CD62p (μg/L): 6.73±1.77 vs. 5.81±1.62, TXB2 (μg/L):18.65±1.77 vs. 17.90±1.68, PF3 (μg/L):5.61±1.48 vs. 4.77±1.24, vWF (μg/L):3.87±1.01 vs. 4.58±1.09, P < 0.05 or P < 0.01]. Conclusion The Sheshang capsule is capable of treating patients with blood coagulative disorder after Trimeresurus stejnegeri snake bite, and its mechanism is possibly related to the improvement of platelet activation function and amelioration of the damage of vascular endothelial cells.

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 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 relationship between indicators of carotid atherosclerosis and onset of ischemic stroke in patients with non-valvular atrial fibrillation (NVAF).Methods:This is a case-control study, a total of 397 NVAF patients with newly diagnosed ischemic stroke (case group) and 3 038 NVAF patients without ischemic stroke (control group) from January 2015 to December 2017 were included in the study. Differences in general clinical features and carotid atherosclerosis indexes between the two groups were compared. Univariate and multivariate logistic regressions were used to analyze the correlation between carotid atherosclerosis indexes and ischemic stroke.Results:Proportions of patients with carotid intima thickening, carotid plaque, stable plaque, unstable plaque, and moderate to severe stenosis were higher in the ischemic stroke group than those in the control group (82.1% vs. 64.4%, 69.3% vs. 50.3%, 43.6% vs. 30.6%, 25.7% vs. 19.7%, and 7.3% vs. 4.0%, respectively, all P <0.05). After adjustment of age, gender, heart failure, hypertension, low density lipoprotein -cholesterol and drug use, multivariate analyses showed that subjects with carotid intima thickening, carotid plaque, stable plaque, unstable plaque, moderate to severe stenosis had 1.766, 2.111, 1.892, 2.256 and 1.824 times the risk for the development of ischemic stroke compared with the subjects without any carotid atherosclerosis indicators. Conclusion:Carotid atherosclerosis, especially with unstable carotid plaque, is associated with ischemic stroke in patients with NVAF.

Objective:To explore the differences of gene expression profiles of precursors of exhausted T cells (Tpex) and terminal exhausted T cells (Tex) in the peripheral blood of patients with active pulmonary tuberculosis (ATB).Methods:Twenty-five cases of ATB, 13 cases of latent tuberculosis infection (LTBI) and 10 health controls were enrolled from January 2021 to October 2022 in the Fifth People′s Hospital of Wuxi. The proportions of Tpex and Tex in the peripheral blood mononuclear cells (PBMCs) of the three groups were detected by flowcytometry. PBMCs of ATB were separated into Tpex and Tex by fluorescence-activated cell sorting. RNA-sequencing was performed and up-regulated and down-regulated genes were screended. Differently expressed genes were analyzed by gene set enrichment analysis of gene ontology (GO) to find regulatory pathways affecting cell metabolism and function. Wilcoxon matched-pairs signed rank test, Kruskal-Wallis test and Dunn multiple comparsion test were used for statistical analysis.Results:The proportion of Tpex in ATB group was 2.86%(1.74%), which was lower than 7.93%(6.16%) of Tex, and the difference was statistically significant ( Z=-3.91, P<0.001). The proportions of Tpex and Tex in LTBI group were 9.47%(6.26%) and 7.43%(5.48%), respectively, and the difference was not statistically significant ( Z=-0.93, P=0.345). The proportions of Tpex and Tex in healthy control group were 8.42%(2.69%) and 6.49%(5.14%), respectively, with no statistical significance ( Z=-1.36, P=0.170). There was statistical difference of the proportion of Tpex among the three groups ( H=21.93, P<0.001), and the proportion of Tpex in ATB group was lower than those in LTBI and heathy control groups, and the differences were both statistically significant ( Z=4.16, P<0.001 and Z=3.34, P=0.003, respectively), while the proportions of Tex in these three groups were not statistically different ( H=2.17, P=0.338). Compared with Tex, the gene expressions of memory markers, such as B-cell lymphoma 2 of Tpex were up-regulated, and the gene expressions of exhausted markers, such as lymphocyte activation gene 3 were down-regulated. In terms of cellular metabolism, the gene expressions of mitochondrial protein complex, mitochondrial matrix and oxidative phosphorylation of Tpex were up-regulated, and the gene expressions of glycolysis were down-regulated. The gene expressions of pyruvate metabolism in Tex were up-regulated, and the gene expressions of CD4 + T lymphocyte activation and differentiation and glycolytic process in Tpex were down-regulated. Conclusions:Tpex in ATB express more characteristics of memory cells and less features of exhausted markers compared with Tex, and the function of mitochondria of Tpex preserves well.