Objective To evaluate the clinical use of the baseline CT angiography(CTA)quantitative score(self-designed collateral circulation quantitative,SD-CCQ)in determining the collateral circulation compensation status in patients with acute ischemic stroke(AIS),as well as the reliability and accuracy of the SD-CCQ score and the Alberta Stroke Program Early CT Score(ASPECTS).Methods Retrospective analysis was made on the clinical and imaging data,including CT,CTA and DWI image data,of 84 patients who were admitted for acute ischemic stroke to the Department of Neurorehabilitation of Zhongshan Hospital of Traditional Chinese Medicine from January 2020 to December 2022.Their CTA source images were annotated using a multi-task deep learning method for vascular segmentation.The ASPECTS score and SD-CCQ score were then applied to the CTA images following vascular segmentation in order to assess the collateral circulation compensation of AIS patients.The Kappa test was used to assess the consistency of the two methods used to assess collateral circulation,and the multifactorial Logistic regression analysis was used to examine the relationship between the SD-CCQ and the prognosis of the AIS patients.Results ASPECTS score had good consistency with SD-CCQ score in evaluating collateral circulation in AIS patients(κ=0.65,P＜0.001),and the diagnostic accuracy of the latter for benign collateral circulation in AIS was 96.15％.Logistic regression analysis showed that the new collateral circulation score,baseline NIHSS,and DWI infarct volume were the main factors affecting the long-term prognosis of AIS patients.Conclusion The new scoring system SD-CCQ can be used to evaluate the compensatory status of collateral circulation in AIS patients,which may help in clinical treatment decision-making and prognosis prediction.

Objective:To investigate the changes of T-lymphocyte subsets, T-cell immunoglobulin and mucin domain molecule-1 (TIM-1) and TIM-3 levels, and cytokines in the peripheral blood of patients with active tuberculosis.Methods:From December 2017 to December 2018, 50 tuberculosis patients and 50 cured tuberculosis patients in Zhejiang Hospital of Integrated Chinese and Western Medicine were selected as the tuberculosis group and cured tuberculosis group, respectively. Fifty healthy individuals in the same period were selected as the control group. Flow cytometry was used to detect the T-lymphocyte subsets in the peripheral blood. The mRNA levels of TIM-1, TIM-3, interferon(IFN)-γ and interleukin(IL)-4 in peripheral blood mononuclear cells (PBMC) were detected by quantitative real-time polymerase chain reacticn (PCR). T test was used for statistical analysis. Results:The ratio of CD4 + /CD8 + T lymphocytes in the tuberculosis group (1.21±0.50) decreased significantly, comparing with those in the cured tuberculosis group (1.88±0.62) and the control group (1.92±0.82). The differences were statistically significant ( t=2.148 and 2.207, respectively, both P<0.05). The mRNA levels of TIM-1, TIM-3 and IL-4 in PBMC in the tuberculosis group were 2.16±0.37, 1.59±0.36 and 1.52±0.69, respectively, which were all higher than those in the cured tuberculosis group (1.60±1.23, 1.01±0.52 and 0.91±0.36, respectively) and the healthy control group (1.40±0.27, 0.92±0.34 and 0.79±0.42, respectively). All of these differences were statistically significant ( t=14.120, 11.440, 17.130, 12.090, 12.050 and 17.030, respectively, all P<0.05). However, the IFN-γ mRNA level (0.43±0.11) was lower than that in the cured tuberculosis group (1.74±0.72) and the control group (1.82±1.17), and the differences were both statistically significant ( t=13.880 and 11.430, respectively, both P<0.05). Conclusion:The immune dysfunction in patients with active tuberculosis may be related to the low ratio of CD4 + /CD8 + T lymphocytes, the increased expressions of TIM-1 and TIM-3, and the imbalance of helper T lymphocyte (Th)1/Th2 cytokines.

Objective To analyze the expression of miRNA-29a in U937 macrophages infected with Mycobacterium tuberculosis and its regulation of target genes.Methods The target genes of miRNA-29a were predicted with softwares miRnada,RNAhybrid and targetscan.The differentiation of U937 macrophages was induced by phorbol ester(PMA), the induced U937 cells were infected with bacilli calmette guerin (BCG).The expression levels of miRNA-29a and its target genes in U937 cells were detected with real-time fluorescence quantitative PCR(RT-fqPCR).The miRNA-29a over-and low-expression U937 macrophage cell lines were constructed and the levels of miRNA-29a and its garget genes were detected.The SPSS 18.0 software was used to analyze the data.Results As predicted by relevant softwares,the miRNA-29a regulate the expression of VEGFA,NFATC3 and TSC22D3 genes.After BCG infection,the expression of miRNA-29a increased to 1.33 fold(P <0.05), and the expression levels of VEGFA, NFATC3 and TSC22D3 were increased to 5.34,99.25 and 2.12 fold,respectively(P<0.01).In the miRNA-29a over-expressing U937 macrophages,the expression levels of VEGFA,NFATC3 and TSC22D3 were up-regulated to 1.35,1.29 and 3.38 fold,respectively(P<0.05 or <0.01).While in the U937 macrophages with miRNA-29a knock-down,the expression levels of VEGFA, NFATC3 and TSC22D3 were down-regulated to 0.07, 0.08 and 0.55 fold, respectively(P <0.01).Conclusion The results suggest that Mycobacterium tuberculosis infection can increase the expression of miRNA-29a in U937 macrophages,further targeting the regulation of VEGFA, NFATC3 and TSC22D3 gene expression, which may participate in the pathogenesis and development of tuberculosis.

Objective To investigate the effects of human dendritic cells ( DCs) infected by bovine Mycobacterium tuberculosis attenuated live bacteria ( BCG) on differentiation of CD4 +naive T cells from neonate cord blood .Methods After infected with BCG , human DCs were cultured with CD 4 +naive T cells from neonate cord blood, the expression of miRNA-99b in DCs and the expression of Foxp3, ROR-γt, IFN-γand IL-10 mRNAs in CD4+ T cells were detected by qRT-PCR.DCs were transfected with miRNA-99b antisense oligonucleotides and co-cultured with neonatal cord blood CD 4 +naive T cells , and the transcription level of CD4 +T cell-related genes was detected .SPSS 15.0 was used to analyze the data .Results The transcriptional activity of miRNA-99b gene in BCG-infected DCs was significantly higher than that in uninfected DCs (t=7.06,P<0.01).Compared with CD4 +T cells co-cultured with uninfected DCs, the mRNA expression of IFN-γ(45.61 ±4.46 vs.3.54 ±1.73, t=32.32, P<0.01), IL-10 (4.17 ±1.06 vs.1.26 ±0.67, t=2.24, P<0.05) in CD4 +T cells co-cultured with BCG-infected DCs was significantly increased, while the mRNA expression of ROR-γt was significantly decreased ( 0.08 ±0.02 vs.0.63 ± 0.10, t=0.42, P<0.01).Compared with CD4 +T cells co-cultured with DCs transfected with NC-siRNA, the miRNA-99b expression was blocked , the mRNA expression of Foxp3 (0.12 ±0.01 vs.1.57 ±0.90, t=1.06, P<0.05), IFN-γ(0.03 ±0.01 vs.0.64 ±0.35, t =0.44, P<0.05), IL-10(0.03 ±0.01 vs. 0.76 ±0.09, t=0.54,P<0.01) in CD4 +T cells was significantly decreased , while the expression ROR-γt mRNA was significantly increased (17.03 ±5.51 vs.1.32 ±0.14, t=11.54,P<0.01).Conclusion BCG induces the imbalance of initial CD 4 +T lymphocytes into Th17/Treg by regulating the expression of miRNA-99b in DCs, leading to the occurrence and development of infection .

Aim To explore the effect of shikonin on stemness maintance of glioma stem cells ( GSCs ). Methods After the U87-MG cells were cultured and isolated, the sphere cells were identified by immuno-fluorescent staining. The alteration of stemness of GSCs by shikonin treatment(2 μmol·L - 1 ) for 12 h, 24 h and 48 h was valued by morphological detection using optical microscope and sub-sphere forming assay. Mo-reover, the related markers of stem cells, such as CD133, were detected in shikonin treated GSCs by western blot assay. Protein expression of PI3K, p-PI3K, Akt and p-Akt was detected by western blot af-ter shikonin treatment alone. Furthermore, by combi-nation with insulin-like growth factor-1 ( IGF-1), we observed the alteration of stemness maintance of shiko-nin-treated GSCs. Results The presence of neural stem cell related markers CD133 and nestin proved the characteristics of GSCs. Shikonin treatment significant-ly inhibited the morphology of GSCs and the sub-sphere forming. Besides, the reduced expression of CD133 was detected in shikonin treated GSCs. Though, the expression of PI3K and Akt did not change compared with the control group, the expression of p-PI3K and p-Akt was reduced. Furthermore, the combination of IGF-1 markedly attenuated the inhibitory effect of shikonin on stemness maintance of GSCs. Conclusion The stemness maintance of GSCs can be significantly inhibited by shikonin treatment, in which PI3K/ Akt pathway is involved.

Objective To discuss the impact of repeated contrast media exposure on renal function in patients who received coronary angiography ( CAG) or percutaneous coronary intervention ( PCI) within 1 week after CTA of coronary ateries. Methods A total of 258 patients who received CAG or PCI after coronary CTA were divided into the study group ( n=132, patients had CAG/PCI within 1 week after CTA) and the control group ( n=126, patients had CAG/PCI 1-2 weeks after CTA). Serum creatinine, cystatin C and estimated GFR were tested before and on day 1, 2 and 3 after procedures. The occurance of contrast-induced nephropathy ( CIN ) was recorded. Resu1ts The baseline clinical characteristics of the patients between the two groups had no significant difference. Preoperative and postoperative serum creatinine, cystatin C and eGFR values on day 1, 2 and 3 had no significant difference between the two groups (all P﹥0. 05). There was no significant difference in the incidence of CIN between two groups (5. 3% in the study group vs. 4. 8% in the control group, P﹥0. 05 ) . Conc1usions It is safe and feasible for patients with eGFR≥60 ml/( min?1. 73 m2 ) to undergo CAG or PCI within 1 week after coronary CTA.

Aim To investigate the signaling mecha-nisms in endothelial monocyte-activating polypeptide-Ⅱ( EMAP-Ⅱ)-induced increase in blood-tumor barri-er ( BTB ) permeability. Methods Relatively pure cerebral microvessel fragments were obtained from the cortex of 3-5 days old Wistar rats by using careful dis-section, enzyme digestion, and dextran centrifugation. Then, these fragments were seeded on dishes and cul-tured primarily. In vitro BTB models were constructed by co-cultivation of rat brain microvascular endothelial cells ( BMECs) with C6 glioma cells. Confluent mono-layers of co-cultured BMECs were divided randomly in-to 5 groups ( each n=6 ): control, EMAP-Ⅱ, H7 +EMAP-Ⅱ, C3 exoenzyme + EMAP-Ⅱ, and C3 ex-oenzyme + H7 + EMAP-Ⅱ groups. Transendothelial electric resistance values and horseradish peroxidase flux were measured to evaluate changes in the BTB permeability . The expression levels of tight junction-re-lated protein occludin and ZO-1 in BMECs were meas-ured by Western blot. Immunofluorescence was used to identify the expression and distribution of occludin and ZO-1 in BMECs. Also, Western blot were used to de-tect the expression levels of myosin light chain ( MLC) and phosphomyosin light chain ( pMLC ) in BMECs. Results Compared with control group, the BTB per-meability of EMAP-Ⅱ group was increased significant-ly. The expression levels of occludin and ZO-1 in BMECs were significantly decreased, accompanied with marked increase in the expression level of pMLC. These above-mentioned effects of EMAP-Ⅱ were sig-nificantly inhibited by pretreatment with H7 ( an inhib-itor of PKC ) or/and C3 exoenzyme ( an inhibitor of RhoA ) . Conclusion Signaling molecules PKC and RhoA play important roles in EMAP-Ⅱ-induced in-crease in BTB permeability; signaling pathways PKC-pMLC and RhoA-pMLC are involved in this process.

Objective To study the effects of Qingjinkangkuoyin (QJKKY) on TNF-α and NE in rats with bronchiectasis.Methods Models were established by intrabronchially injecting with pseudomonas aeruginosa,and divided into 5 groups by random:the QJKKY high dose treatment group (given high dose of QJKKY into stomach),the QJKKY low dose treatment group (given low dose of QJKKY),the levofloxacin group (given levofloxacin),the model group (given normal saline),and the normal contrast group (given normal saline).After 2 weeks of treatment,the histopathology of lung tissue,the levels of TNF-α and inflammatory cells in peripheral blood and NE in rats' lung tissue were detected.Results Compared with the model group (160.425±9.9293)ng/L,QJKKY could decrease the level of TNF-α in blood significantly [high dose of QJKKY treatment group was (137.133±6.1646)ng/L,P＜0.05]; the expression of inflammatory cells in serum were decreased significantly by QJKKY [high dose of QJKKY treatment group was (1.106± 0.3580) 109/L,P＜0.05].Low dose of QJKKY treatment group was (1.086 ±0.2433) 109/L,(P＜0.05) ; the expression of NE in lung tissue were decreased remarkably by QJKKY [high dose of QJKKY treatment group(80.697 ±4.5877)ng/L,P＜0.05]; low dose of QJKKY treatment group is (80.747±3.6925)ng/L,(P＜0.05); and the histopathologic change of lung tissue in QJKKY treatment groups were ameliorated under light microscope by HE staining.Conclusion Qingjinkangkuoyin could cure bronchiectasis by decreasing the expression of TNF-αin peripheral blood and NE in rats' lung tissue.

Objective To observe the efficacy and safety of different initial doses of nebulized budesonide inhalation (BI) in infants and young children with moderate to severe asthma exacerbations.Methods A multi-center,parallel controlled clinical trial was performed during Sep 2008 to Apr 2010 in three hospitals,which were Department of Pediatrics,Shanghai Jiaotong University Affiliated Shanghai First People's Hospital,Department of Pediatrics,Shanghai Jiaotong University School of Medicine Affiliated Xinhua Hospital,and Department of Respiratory,Fudan University Affiliated Children's Hospital.One hundred and fifty children aged 6 to 36 month with moderate to severe asthma exacerbations were randomly divided into two groups.The high-starting-dose group was treated with a dose of 1 mg nebulized BI every 8 h for 2 days,while the conventional-starting-dose group was treated with a dose of 0.5 mg cvcry 8 h for 4 days.The terbutaline sulfate aerosol liquid was administered with a dose of 2.5 mg each time as needed.The primary outcome measures were severity scores,which were assessed at admission (0 h),and 8 h,16 h,24 h,48 h,72 h after treatment separately.The secondary outcome measures included the use of β2 receptor agonist,the systemic use of corticosteroids,average length of hospital stay and total cost.The data was analyzed with SPSS 13.0.Results (1) The clinical severity scores were significantly decreased at all time points after treatment in both groups (P ＜ 0.05).Compared with conventional starting-dose of BI,high starting-dose of 3.25 ± 1.82,P ＜ 0.01).(2) The terbutaline doses and the systemic corticosteroids do-ses were significantly reduced in high-starting-dose group compared with conventional-starting-dose group [(16.27 ± 12.99) mg vs (22.90 ± 18.27) mg,P ＜ 0.05 ; (4.54 ± 18.18) mg vs (11.16 ± 21.34) mg,P ＜ 0.05).The average length of hospital stay and the total cost of the two groups showed no significant differences (P ＞ 0.05).(3) There were no side effects associated with BI.Conclusion Compared with conventional treatment,high-starting-dose of BI can control symptoms fast and reduce the use of systemic corticosteroid without any side effects.BI improved symptoms more quickly at 8 h (2.87 ± 1.60 vs 4.48 ± 2.24,P ＜ 0.01) and 16 h (2.48 ± 1.56 vs