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Objective:To observe the expression levels of Toll-like receptor 4 (TLR4) signaling pathway-related proteins and their phosphorylation in the liver tissues of rats with inorganic arsenic poisoning, and to explore the role of TLR4-mediated inflammatory signaling pathway in arsenic-induced liver fibrosis injury.Methods:Eighteen healthy weanling SD rats were divided into 3 groups according to their body weight (80 - 100 g) using a random number table (6 rats in each group, half males and half females). The control group was given 10 ml/kg of normal saline by gavage. The sodium arsenite (NaAsO 2) exposure group was given 10 mg/kg of NaAsO 2 by gavage. The TAK-242 intervention group was given 10 mg/kg of NaAsO 2 by gavage, and 0.5 mg/kg of TAK-242 was also administered intraperitoneally to inhibit TLR4 after 12 weeks. All rats were administered 6 days a week for 36 weeks. At the end of the treatment, the liver tissues and serum of the rats in each group were collected. HE and Masson staining were used to observe the pathological and fibrotic changes of the liver tissues. Automatic biochemical analyzer was used to detect serum liver function indexes of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP). Western blot was used to detect the expression changes of rat liver fibrosis protein α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), Vimentin and TLR4 signaling pathway-related proteins TLR4, nuclear factor κB (NF-κB)-p65 subunit (p65), NF-κB-p50 subunit (p50) and their phosphorylation p-p65 and p-p50 expression levels. Enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion levels of inflammatory related factors interleukin (IL)-6, tumor necrosis factor-α (TNF-α) and IL-10. Results:HE and Masson staining results showed that compared with the control group, the NaAsO 2 exposure group showed significant inflammatory cell infiltration, hepatocyte necrosis and collagen fibrous deposition, while the TAK-242 intervention group showed improvement of the inflammatory cell infiltration and reduction of collagen fibrous deposition compared with the NaAsO 2 exposure group. The results of serum liver function indexes showed that ALT, AST and ALP in NaAsO 2 exposure group were increased compared with the control group, but the TAK-242 intervention group was significantly decreased compared with the NaAsO 2 exposure group ( P < 0.05). Western bolt results showed that in NaAsO 2 exposure group, the expression levels of fibrosis protein α-SMA, TGF-β1 and Vimentin (1.04 ± 0.19, 0.92 ± 0.14, 1.20 ± 0.21) and TLR4 signaling pathway-related proteins and their phosphorylation TLR4, p50, p-p50 and p-p65 (1.16 ± 0.21, 0.95 ± 0.16, 1.24 ± 0.23, 1.56 ± 0.25) were higher than the control group (0.44 ± 0.08, 0.42 ± 0.08, 0.72 ± 0.07, 0.69 ± 0.15, 0.71 ± 0.11, 0.46 ± 0.07, 0.54 ± 0.11, P < 0.05), and the TAK-242 intervention group (0.60 ± 0.13, 0.59 ± 0.16, 0.49 ± 0.11, 0.47 ± 0.08, 0.86 ± 0.09, 0.79 ± 0.14, 1.02 ± 0.17) were lower than the NaAsO 2 exposure group ( P < 0.05). There was no significant difference in the expression level of TLR4 signal pathway-related protein p65 among the three groups ( F = 14.29, P = 0.053). ELISA results showed that the secretion levels of IL-6 and TNF-α [(98.89 ± 4.58), (83.25 ± 4.57) ng/g] in rats liver tissues of the NaAsO 2 exposure group were higher than the control group [(27.30 ± 3.92), (27.77 ± 1.83) ng/g, P < 0.05], while the secretion level of IL-10 [(36.88 ± 3.86) ng/g] was lower than the control group [(77.96 ± 7.87) ng/g, P < 0.05]. In TAK-242 intervention group, IL-6 and TNF-α secretion levels [(44.32 ± 3.60), (36.51 ± 2.93) ng/g] were lower and IL-10 secretion level [(60.40 ± 4.94) ng/g] was higher compared with the NaAsO 2 exposure group ( P < 0.05). Conclusion:TLR4-mediated inflammatory signaling pathway-related proteins and their phosphorylation are highly expressed in the liver tissues of rats with inorganic arsenic poisoning, and inhibition of TLR4 signaling pathway could significantly reduce the degree of liver fibrosis injury caused by inorganic arsenic in rats.
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OBJECTIVE@#To validate the effectiveness of a novel comprehensive classification for intertrochanteric fracture (ITF).@*METHODS@#The study included 616 patients with ITF, including 279 males (45.29%) and 337 females (54.71%); the age ranged from 23 to 100 years, with an average of 72.5 years. Two orthopaedic residents (observers Ⅰ and Ⅱ) and two senior orthopaedic surgeons (observers Ⅲ and Ⅳ) were selected to classify the CT imaging data of 616 patients in a random order by using the AO/Orthopaedic Trauma Association (AO/OTA) classification of 1996/2007 edition, the AO/OTA classification of 2018 edition, and the novel comprehensive classification method at an interval of 1 month. Kappa consistency test was used to evaluate the intra-observer and inter-observer consistency of the three ITF classification systems.@*RESULTS@#The inter-observer consistency of the three classification systems evaluated by 4 observers twice showed that the 3 classification systems had strong inter-observer consistency. Among them, the κ value of the novel comprehensive classification was higher than that of the AO/OTA classification of 1996/2007 edition and 2018 edition, and the experience of observers had a certain impact on the classification results, and the inter-observer consistency of orthopaedic residents was slightly better than that of senior orthopaedic surgeons. The intra-observer consistency of two evaluations of three classification systems by 4 observers showed that the consistency of the novel comprehensive classification was better for the other 3 observers, except that the consistency of observer Ⅳ in the AO/OTA classification of 2018 version was slightly higher than that of the novel comprehensive classification. The results showed that the novel comprehensive classification has higher repeatability, and the intra-observer consistency of senior orthopaedic surgeons was better than that of orthopaedic residents.@*CONCLUSION@#The novel comprehensive classification system has good intra- and inter-observer consistency, and has high validity in the classification of CT images of ITF patients; the experience of observers has a certain impact on the results of the three classification systems, and those with more experiences have higher intra-observer consistency.
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Male , Female , Humans , Young Adult , Adult , Middle Aged , Aged , Aged, 80 and over , Observer Variation , Reproducibility of Results , Hip Fractures/surgery , Tomography, X-Ray Computed/methods , RadiographyABSTRACT
Objective To evaluate the effect of body mass index (BMI) on early prognosis of the recipients after lung transplantation. Methods Clinical data of 307 lung transplant recipients were retrospectively analyzed. According to preoperative BMI, all recipients were divided into the low (BMI <18.5 kg/m2, n=114), normal (18.5 kg/m2≤BMI <24 kg/m2, n=145) and high (BMI≥24.0 kg/m2, n=48) BMI groups, respectively. Baseline data including age, sex, blood type, BMI, preoperative complications, preoperative pulmonary hypertension, and intraoperative use of extracorporeal membrane oxygenation (ECMO) of all recipients were compared among three groups. The survival rate of all recipients was estimated by Kaplan-Meier curve and the survival curve was delineated. The differences of survival rate were analyzed by log-rank test. The 30-, 90- and 180-d mortality risk of all recipients after lung transplantation in different BMI groups was compared by multivariate Cox regression analysis. Results There were significant differences in age and sex of recipients among three groups (both P<0.05). There was a significant difference regarding the 180-d survival rate after lung transplantation among different BMI groups (P<0.05). Multivariate Cox regression analysis showed that the 90-d mortality risk after lung transplantation in the high BMI group was 2.295 times higher than that in the normal BMI group [hazard ratio (HR) 2.295, 95% confidence interval (CI) 1.064-4.947, P=0.034]. In the high BMI group, the 180-d mortality risk after lung transplantation was 2.783 times higher compared with that in the normal BMI group (HR 2.783, 95%CI 1.333-5.810, P=0.006), and the 180-d mortality risk in the low BMI group was 2.181 times higher than that in the normal BMI group (HR 2.181, 95%CI 1.124-4.232, P=0.021). Conclusions Compared with the recipients with normal BMI, their counterparts with high and low preoperative BMI have higher mortality risk early after lung transplantation. Adjusting preoperative BMI to normal range contributes to improving early prognosis of lung transplant recipients.
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The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in more than 235 million cases worldwide and 4.8 million deaths (October 2021). Severe COVID-19 is characterised in part by vascular thrombosis and a cytokine storm due to increased plasma concentrations of factors secreted from endothelial and T-cells. Here, using patient data recorded in the UK Biobank, we demonstrate the importance of variations in Rab46 (CRACR2A) with clinical outcomes. Using logistic regression analysis, we determined that three single nucleotide polymorphisms (SNPs) in the gene EFCAB4B cause missense mutations in Rab46, which are associated with COVID-19 fatality independently of risk factors. All three SNPs cause changes in amino acid residues that are highly conserved across species, indicating their importance in protein structure and function. Two SNPs, rs17836273 (A98T) and rs36030417 (H212Q), cause amino acid substitutions in important functional domains: the EF-hand and coiled-coil domain respectively. By using molecular modelling, we suggest that the substitution of threonine at position 98 causes structural changes in the EF-hand calcium binding domain. Since Rab46 is a Rab GTPase that regulates both endothelial cell secretion and T-cell signalling, these missense variations may play a role in the molecular mechanisms underlying the thrombotic and inflammatory characteristics observed in patients with severe COVID-19 outcomes.
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Background Long-term exposure to sodium arsenite leads to its accumulation in the liver and liver injury as a result. Previous studies showed that mesenchymal cells play an important role in hepatic fibrosis, and epithelial-mesenchymal transformation (EMT) is considered to be a main source of mesenchymal cells. Objective To investigate the effects of sodium arsenite at different doses on liver fibrosis and EMT-related protein expressions in SD rats. Methods Twenty-four healthy weaned SD rats, half male and half female, were randomly divided into four groups according to body weight, with 6 rats in each group. The four groups were control group (gavage with 10.0 mL·kg−1 physiological saline), 2.5 mg·kg−1 sodium arsenite group, 5.0 mg·kg−1 sodium arsenite group, and 10.0 mg·kg−1 sodium arsenite group. All rats were gavaged 6 d per week for 36 weeks and weighed once a week, the serum and liver tissues of rats were collected and weighed, then the organ coefficient was calculated. Hematoxylin-eosin staining and Masson's trichrome staining were used to determine the pathological changes of hepatic fibrosis in rats. The serum secretion levels of hyaluronic acid (HA), laminin (LN), procollagen Ⅲ N-terminal propeptide (PⅢNP), and collagen Ⅳ (COL-Ⅳ) in rats were detected by enzyme-linked immunosorbent assay (ELISA). The protein expressions of HSCs activation-related proteins, such as α-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1), as well as EMT-related markers, such as E-cadherin, N-cadherin, Vimentin, and Snail, were detected by Western blotting. Results Compared with the control group, the 10.0 mg·kg−1 sodium arsenite group showed decreased body weight (P<0.05) and increased liver coefficient (P<0.05) of female and male rats. The pathological staining showed that, compared with the control group, a large number of inflammatory cells were observed in liver tissue of rats exposed to sodium arsenite, liver parenchymal cells were also liquefied, necrotic, and denatured, and the collagen positive staining area of liver tissue showed an upward trend along with the increase of arsenic exposure dose (P<0.05). The results of ELISA and Western blotting showed that the serum secretion levels of HA, LN, PⅢNP, and COL-Ⅳ in the 5.0 and 10.0 mg·kg−1 sodium arsenite groups were higher than those in the control group and the 2.5 mg·kg−1 sodium arsenite group (P<0.05). Compared with the control group, the expressions of α-SMA and TGF-β1 proteins in liver tissue were increased in each sodium arsenite exposure group (P<0.05), the expression levels of E-cadherin protein were decreased (P<0.05), and the expression levels of N-cadherin, Vimentin, and Snail were increased (P<0.05). Conclusion Sodium arsenite exposure can induce HSCs activation and liver fibrosis injury in SD rats, resulting in increased extracellular matrix secretion levels, accompanied by EMT in liver tissue, suggesting that EMT is closely related to the process of liver fibrosis caused by arsenic.
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Objective:To investigate the effect of voglibose combined with exenatide on blood glucose control and serum fibroblast growth factor 21 (FGF-21) expression in patients with type 2 diabetes mellitus (T2DM).Methods:Three hundred and eighty patients with T2DM admitted to Qingdao Municipal Hospital from October 2018 to September 2020 were selected and randomly divided into the control group and the observation group, with 190 patients in each group. The control group was treated with voglibose, and the observation group was treated with voglibose and exenatide. Both groups were treated for 16 weeks. Blood glucose, blood fat, insulin and other indicators as well as serum FGF-21 levels were compared between the two groups before and after the treatment, and the efficacy of drugs and the incidence of adverse reactions were compared.Results:The total effective rate in the observation group was higher than that in the control group: 97.89%(186/190) vs. 93.16%(177/190), χ2 = 5.00, P<0.05. After treatment, the levels of fasting plasma glucose (FPG), 2 h plasma glucose (2 h PG) and glycosylated hemoglobin (HbA 1c) in the observation group were lower than those in the control group: (6.95 ± 1.03) mmol/L vs. (8.29 ± 1.15) mmol/L, (7.88 ± 2.07) mmol/L vs. (10.03 ± 3.24) mmol/L, (7.17 ± 1.08)% vs. (8.13 ± 1.21)%, P<0.05. After treatment, the levels of triacylglycerol (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in the observation group were lower than those in the control group: (2.13 ± 0.23) mmol/L vs. (2.93 ± 0.34) mmol/L, (3.10 ± 1.01) mmol/L vs. (3.98 ± 1.14) mmol/L, (1.93±0.38) mmol/L vs. (2.73±0.54) mmol/L, P<0.05. After treatment, the level of islet β cell function index (HOMA-β) in the observation group was higher than that in the control group: 111.56 ± 20.78 vs. 102.23 ± 20.14, P<0.05. After treatment, the level of serum FGF-21 in the observation group was lower than that in the control group: (142.09 ± 26.82) ng/L vs. (150.22 ± 30.21) ng/L, P<0.05. The incidence of adverse reactions between the two groups had no significant difference ( P>0.05). Conclusions:Voglibose combined with exenatide has a definite effect on T2DM patients with metformin failure, which can effectively control their blood glucose level, reduce serum FGF-21 level, improve insulin resistance and glucose and lipid metabolism, and reduce body mass, with high safety.
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Background Arsenic is a well-known environmental toxicant. Hepatic fibrosis could occur dueto excessive or long-term exposure to arsenic, while associated molecular mechanisms remain undefined. Mitogen-inducible gene 6 (Mig-6) exhibits a protective effect on numerous diseases or cancers. However, the specific role of Mig-6 in the mechanisms of arsenite-induced hepatic fibrosis remains indistinct. Objective To investigate the specific role of Mig-6 in the activation of hepatic stellate cells (HSC) and the deposition of extracellular matrix (ECM) induced by sodium arsenite (NaAsO2). Methods Human hepatic stellate cells (Lx-2) were treated with 0, 1.875, 3.75, 7.5, and 15 μmol·L−1 of NaAsO2 for 24 h, or with 7.5 μmol·L−1 NaAsO2 for 0, 12, 24, 48, and 72 h. Additionally, Lx-2 cells were transfected by pcDNA3.1(+)/Mig-6, then treated with 7.5 μmol·L−1 NaAsO2 for 24 h; a blank control group, a pcDNA3.1(+)-control group, a pcDNA3.1(+)/Mig-6 group, and an arsenic (7.5 μmol·L−1 NaAsO2) group were also set up. After transfection, the cells and culture supernatants were collected, and the protein levels of Mig-6, α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) in Lx-2 cells were identified by Western blotting analysis; moreover, the secretion levels of main ECM components in supernatants such as hyaluronic acid (HA), laminin (LN), collagens IV (COL-IV), and procollagen-III (PIIINP) were tested by ELISA. Results The Mig-6 expression decreased in the 3.75, 7.5, and 15 μmol·L−1 NaAsO2 groups (0.561±0.095, 0.695±0.048, and 0.401±0.030) compared to the control group (1.000±0.000) in Lx-2 cells (P<0.05). After administration with 7.5 μmol·L−1 of NaAsO2 for 24, 48, and 72 h, the Mig-6 expression (0.856±0.036, 0.515±0.077, 0.491±0.060) decreased compared with the 0 h group (1.000±0.000) (P<0.05). After over-expression of Mig-6, the results of Lx-2 activation related protein levels showed that compared to the control group, the α-SMA and TGF-β1 expression were up-regulated in the arsenic group (P<0.05); meanwhile, the α-SMA and TGF-β1 in the Mig-6 over-expression combined arsenic exposure group reduced compared to the arsenic (7.5 μmol·L−1) group (P<0.05). The results of ELISA showed that compared with the control group, the HA, LN, PIIINP, COL-IV in the arsenic group were up-regulated (P<0.05); while compared to the arsenic group, the HA, LN, PIIINP, and COL-IV in the Mig-6 over-expression combined with arsenic exposure group were decreased (P<0.05). Conclusion Arsenic down-regulates Mig-6 expression in HSC, and over-expression of Mig-6 can reverse the activation of HSC and ECM deposition induced by arsenic exposure. It suggests that Mig-6 plays a protective role in arsenic-induced HSC activation and ECM deposition.
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Treatment of severe COVID-19 is currently limited by clinical heterogeneity and incomplete understanding of potentially druggable immune mediators of disease. To advance this, we present a comprehensive multi-omic blood atlas in patients with varying COVID-19 severity and compare with influenza, sepsis and healthy volunteers. We identify immune signatures and correlates of host response. Hallmarks of disease severity revealed cells, their inflammatory mediators and networks as potential therapeutic targets, including progenitor cells and specific myeloid and lymphocyte subsets, features of the immune repertoire, acute phase response, metabolism and coagulation. Persisting immune activation involving AP-1/p38MAPK was a specific feature of COVID-19. The plasma proteome enabled sub-phenotyping into patient clusters, predictive of severity and outcome. Tensor and matrix decomposition of the overall dataset revealed feature groupings linked with disease severity and specificity. Our systems-based integrative approach and blood atlas will inform future drug development, clinical trial design and personalised medicine approaches for COVID-19.
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Objective:To compare clinical effects of the open versus closed high tibial osteotomy on knee osteoarthritis.Methods:A total of 100 patients with knee osteoarthritis admitted to our hospital from May 2018 to May 2019 were included.They were randomly divided into groups A and B(n=50, each group)according to the principle of random and double blind.Patients in group A received the medial opening high tibial osteotomy, and group B were treated with lateral closed high tibial osteotomy.The changes in the Lysholm knee score, hospital for special surgery(HSS)knee score and complications were compared between the two groups before and 1 year after surgery.The correction angle, the change of patella height before and after operation, and the change of posterior slope of tibial plateau were compared between the two groups.Results:Before and after treatment, Lysholm scores were(63.51±5.47)and(90.98±5.84)( t=24.275, P=0.000), and HSS scores were(51.85±4.68)and(88.64±5.87)( t=34.652, P=0.000). Lysholm scores were(62.98±6.14)and(91.52±6.54 9)( t=22.497, P=0.000), and HSS scores were(52.05±5.16)and(89.54±5.15)( t=36.362, P=0.000)in group A and B, .After treatment, all index were significantly improved in the two groups, but there was no statistical difference between the two groups( P>0.05). In group A, the posterior slope of tibial plateau were(8.75±1.48)° and(10.25±1.65)°( t=4.785, P=0.000)and the patellar height was(0.890±0.031)and(0.898±0.032)( t=1.270, P=0.207)before and after treatment.Before and after treatment in group B, the posterior slope of tibial plateau were(8.69±1.53)° and(5.26±1.21)°( t=12.434, P=0.000)and the patellar height were(0.889±0.047)and(0.821±0.039)( t=7.873, P=0.000). The correction angle, posterior slope of tibial plateau and patella height were significantly improved after treatment in the two groups.While, the decreases of posterior slope of tibial plateau and patella height were better in the group B than in the group A( P<0.05). There was no significant difference in the incidence of complications between the two groups( P>0.05). Conclusions:For treatment of knee osteoarthritis patients, the medial opening high tibial osteotomy and lateral closed high tibial osteotomy have the same exact effect and high safety, but the two methods have their own advantages and disadvantages in clinical treatment.And the appropriate surgical treatment can be selected according to the characteristics of patients.
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Objective:To investigate the main postoperative complications, causes of death and the risk factors for survival in patient with benign end-stage lung diseases within 1 year after lung transplantation.Methods:A retrospective analysis was conducted to collect the clinical data of 200 patients with benign end-stage lung disease who underwent lung transplantation admitted to Wuxi People's Hospital Affiliated to Nanjing Medical University from May 2017 to October 2018. The main postoperative complications, survival and causes of death within 1 year after operation were analyzed. The Kaplan-Meier method was used to plot survival curves, and the Log-Rank test was used to compare the influence of factors, including recipient's gender, use of marginal donor lung, primary disease, preoperative combination of moderate to severe pulmonary hypertension (PAH), intraoperative extracorporeal membrane oxygenation (ECMO) support, surgical methods, intraoperative massive blood loss, postoperative complications [infection, primary graft dysfunction (PGD), acute rejection], on 1-year survival in patients who underwent lung transplantation. The multivariate Cox proportional hazards regression model was used to evaluate the risk factors of death within 1 year after lung transplantation.Results:Two hundred patients underwent successful lung transplantation. The major postoperative complications within 1 year after transplantation included infection in 131 patients, PGD in 20 patients, acute rejection in 57 patients, anastomotic complication in 26 patients and others (new onset diabetes, osteoporosis, etc.) in 53 patients. The 3-month, 6-month, and 1-year postoperative cumulative survival rates were 81.5%, 80.0% and 77.5%, respectively. Forty-five patients died during 1 year after operation, among whom 14 died of infection, 7 died of PGD, 8 died of acute rejection, 4 died of anastomotic complication, 3 died of cardio-cerebrovascular accident, 3 died of multiple organ failure, 2 died of respiratory failure and 4 died of other causes (traffic accident, etc.). The Kaplan-Meier survival analysis showed that recipient's gender, idiopathic pulmonary fibrosis (IPF) as the primary disease, preoperative combination of moderate and severe PAH, intraoperative ECMO support, intraoperative massive blood loss, postoperative complications (infection, PGD, acute rejection) were influencing factors for postoperative 1-year survival rate. The multivariate Cox regression model showed that male was the protective factor [hazard ratio ( HR) = 0.481, 95% confidence interval (95% CI) was 0.244-0.947, P = 0.034], IPF as the primary disease ( HR = 2.667, 95% CI was 1.222-5.848, P = 0.014), intraoperative use of ECMO support ( HR = 1.538, 95% CI was 0.787-3.012, P = 0.028), massive blood loss during surgery ( HR = 2.026, 95% CI was 0.976-4.205, P = 0.045) and postoperative infection ( HR = 3.138, 95% CI was 1.294-7.608, P = 0.011), PGD ( HR = 1.604, 95% CI was 0.464-5.539, P = 0.004), and acute rejection ( HR = 1.897, 95% CI was 0.791-4.552, P = 0.015) were the independent risk factors for death within 1 year after transplantation. Conclusions:One-year survival rates after lung transplantation are affected by recipient's gender, primary disease, preoperative combination of moderate and severe PAH, intraoperative ECMO support, intraoperative massive blood loss, and postoperative complications (infection, PGD, acute rejection). The male is the protective factor, while IPF as the primary disease, intraoperative ECMO support, massive blood loss during surgery and postoperative complications (infection, PGD, acute rejection) are independent risk factors for death within 1 year after lung transplantation.
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Objective:To investigate the effect of propranolol on angiopoietin and its receptor in the nude mouse model of infantile hemangioma, so as to clarify the mechanism of propranolol in the treatment of hemangioma.Methods:The proliferative hemangioma tissue from the 980 Hospital of PLA was transplanted into the back of 50 female nude mice during May 2015 to Sept. 2016. After establishment of the hemangioma model, the nude mice were randomly divided into two groups, 25 in each group. The experimental group was gavaged with 0.4 ml propranolol every 2 days, and the control group was gavaged with normal saline every 2 days. The mice were killed in batches on the 7th, 14th, 21st and 28th day, the tumor morphology was observed by HE staining. The expression of VEGF, Ang1, Ang2 and Tie2 protein in hemangiomas was detected by immunohistochemistry and Western blot.Results:After 28 days, the hemangioma in propranolol group showed regression phase. The endothelial cells of hemangioma decreased, and the nuclear staining became shallow, the lumen enlarged, and fibrous connective tissue could be seen between the blood vessels. The expression of VEGF, Ang2 and Tie2 was lower than those in the control group ( P<0.05), and the expression of VEGF, Ang2 and Tie2 was decreased by 47.1%, 34.7% and 37.5%, respectively, while the expression of Ang1 was increased by 40.5% compared with the control group ( P<0.05). Conclusions:Propranolol may inhibit the growth of hemangioma in nude mice by promoting the expression of Ang1 and inhibiting the expression of VEGF, Ang2 and Tie2.
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Public health emergency of SARS-CoV-2 has facilitated diagnostic testing as a related medical countermeasure against COVID-19 outbreak. Numerous serologic antibody tests have become available through an expedited federal emergency use only process. This paper highlights the analytical characteristic of an ELISA based assay by AnshLabs and three random access immunoassay (RAIA) by DiaSorin, Roche, and Abbott that have been approved for emergency use authorization (EUA), at a tertiary academic center in a low disease-prevalence area. The AnshLabs gave higher estimates of sero-prevalence, over the three RAIA methods. For positive results, AnshLabs had 93.3% and 100% concordance with DiaSorin or Abbott and Roche respectively. For negative results, AnshLabs had 69.7% and 73.0% concordance with DiaSorin and Roche or Abbott respectively. All discrepant samples that were positive by AnshLabs and negative by RAIA tested positive by all-in-one step SARS-CoV-2 Total (COV2T) assay performed on the automated Siemens Advia Centaur XPT analyzer. None of these methods, however, are useful in early diagnosis of SARS-CoV-2.
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Objective:To improve awareness about infratentorial dural arteriovenous fistula (DAVF).Methods:Three cases of DAVF in the First Affiliated Hospital of Soochow University from September 2017 to September 2019 were retrospectively analyzed in terms of clinical features, cerebrospinal fluid (CSF) analysis, brain imaging and treatment, and followed up through telephone call.Results:Case 1: A 43-year-old woman, in chronic but acute aggravated course, presented with weakness of both lower limbs and urination and defecation dysfunction. Brain magnetic resonance imaging (MRI) revealed abnormal signal in medulla. CSF analysis demonstrated aquaporin-4 antibodies positive. Misdiagnosed as neuromyelitis optica spectrum disorders, the treatment was poor. Then digital subtraction angiography (DSA) showed DAVF at the left infratentorial area, and endovascular treatment was operated. Relapse was not observed in two-year follow up. Case 2: A 57-year-old woman, in chronic progressive course, mainly manifested as memory loss, but progressed with dysphagia, fever, coma. Treatment as “central nervous infection” was poor. Then DSA showed DAVF at the bilateral transverse-sigmoid sinus area, and endovascular treatment was operated with embolized partial fistulas. The patient died from lung infection within two months. Case 3: A 52-year-old man, in subacute course, was treated in the Gastroenterology Department with clinical manifestion of stubborn nausea and vomiting. Brain MRI revealed abnormal signal in medulla, with prominent vessel flow voids nearby. Then DSA showed DAVF at the craniocervical junction, and endovascular treatment was operated. Relapse was not observed in six-month follow up.Conclusions:DAVF has a variety of clinical manifestations, and infratentorial DAVF can manifest as acute neurological dysfunction involving the brain stem, cerebellum, spinal cord, which may be easily misdiagnosed. When brain MRI showed intracranial abnormal signal, the possibility of DAVF should be considered. DSA remains the gold standard to diagnose DAVF. Endovascular embolization is the main treatment of infratentorial DAVF at present. Prognosis depends on clinical presentation and fistula classification.
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Objective To investigate the expression of transcription factor forkhead/winged helix transcription factor 3 (Foxp3),immune factor transforming growth factor-beta 1 (TGF-β1),and T-lymphocyte activation related factor interleukin-2 (IL-2) in peripheral blood of patients with coal-burning arsenic poisoning,and to analyze the effects of arsenic exposure on immune function.Methods A case-control study was conducted to investigate 149 cases [94 males and 55 females,(50.69 ± 6.14) years old] of arsenic poisoning in Yuzhang coalburning arsenic poisoning area,southwestern Guizhou Province,and the cases were diagnosed based on the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and confirmed by clinical review.According to skin damage,the patients were divided into mild arsenic poisoning group (39 cases),moderate arsenic poisoning group (54 cases) and severe arsenic poisoning group (56 cases);and 41 cases [12 males and 29 females,(45.76 ± 7.88) years old] of non-arsenic exposed residents from 12 km of Yuzhang coal-burning area were selected as control group.Morning urine and peripheral blood samples were collected with informed consent.Urine arsenic content was measured by inductively coupled plasma mass spectrometry (ICP-MS).Urine arsenic was corrected by creatinine (Cr).Detection of regulatory T cell (Treg)-specific transcription factor Foxp3 gene expression in human peripheral blood was done by real-time fluorescence quantitative PCR,and the levels of Treg-related immune factor TGF-β1 and IL-2 in serum were detected by enzyme linked immunosorbent assay (ELISA).Results The urinary arsenic contents [median (quartile):29.13 (19.75-54.50),31.81 (17.52-53.31),30.51 (18.35-45.76) μg/g Cr] in each arsenic poisoning group were higher than that in the control group [21.62 (17.65-28.44) μg/g Cr,P < 0.05].The expression levels of Foxp3 mRNA in peripheral blood of each arsenic poisoning group [median (quartile):0.58 (0.17-1.27),0.32 (0.17-0.61),0.33 (0.13-0.62)] were significantly lower than that in the control group [0.87 (0.64-1.50),P < 0.05];compared with mild arsenic poisoning group,the expression of Foxp3 mRNA in peripheral blood of moderate and severe arsenic poisoning groups decreased (P < 0.05).The contents of serum TGF-β1 [(13.14 ± 5.19),(12.85 ± 5.51),(12.78 ± 4.95) μg/L] in each arsenic poisoning group were significantly higher than that in the control group [(3.90 ± 1.53) μg/L,P < 0.05].The levels of IL-2 in serum of each arsenic poisoning group [(9.85 ± 5.38),(11.64 ± 6.40),(12.27 ± 6.19) ng/L] were lower than that in the control group [(34.30 ± 4.84) ng/L,P < 0.05];the serum level of IL-2 in severe arsenic poisoning group was higher than that in mild arsenic poisoning group (P < 0.05).Conclusions Arsenic exposure can cause abnormal changes of Treg-specific transcription factor Foxp3 and related immune factors TGF-β1 and IL-2 in peripheral blood of patients.It is suggested that Treg dysfunction may be related to arsenic poisoning.
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Objective To observe the change of T helper 17 (Th17),regulatory T cell (Treg) as a percentage of lymphocytes and the Th17/Treg ratio in peripheral blood of patients with coal-burning-borne arsenic poisoning,and to explore the role of Th17 cells and Treg cells balance in arsenic-induced immune injury.Methods A case-control study was conducted to investigate 149 cases of arsenic poisoning in Yuzhang arsenic poisoning area in the southwestern Gnizhou Province,and the age was (50.69 ± 6.14) years old,including 94 males and 55 females.The diagnosis was based on the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015),and the cases were divided into mild arsenic poisoning group (39 cases),moderate arsenic poisoning group (54 cases) and severe arsenic poisoning group (56 cases);forty--one residents of non-arsenic exposed villages about 12 km away from the diseased area were collected as control group,the age was (45.76 ± 7.88) years old,including 12 males and 29 females.Hair samples and peripheral blood were collected from the subjects.The content of hair arsenic was detected by inductively coupled plasma mass spectrometry (ICP-MS).The percentages of Th17 cells and Treg cells in peripheral blood lymphocytes were detected by flow cytometry,and changes in the ratio of Th17/Treg in each group were analyzed.Results The hair arsenic contents in control,mild,moderate,and severe arsenic poisoning groups [median (quartile)] were 0.12 (0.08-0.18),0.20 (0.16-0.33),0.25 (0.18-0.41),0.28 (0.21-0.50) μg/g,and the differences were statistically significant between groups (H =52.22,P < 0.01),and the hair arsenic content in each arsenic poisoning group was higher than that of the control group (P < 0.05).The percentages of Th17 cells in peripheral blood lymphocytes of moderate and severe arsenic poisoning groups [(0.42 ± 0.21)%,(0.41 ± 0.23)%] were higher than that of the control group [(0.29 ± 0.16)%,P < 0.05].The percentages of Treg cells in peripheral blood lymphocytes of mild,moderate and severe arsenic poisoning groups [(0.37 ± 0.18)%,(0.31 ± 0.19)%,(0.27 ± 0.18)%] were lower than that of the control group [(0.71 ± 0.20)%,P < 0.05];with respect to the mild arsenic poisoning group,the percentage of Treg cells in severe arsenic poisoning group was reduced (P < 0.05).The ratios of Th17/Treg in mild,moderate and severe arsenic poisoning groups (1.17 ± 0.63,1.56 ± 0.69,1.83 ± 0.85) were higher than that of the control group (0.43 ± 0.22,P < 0.05);compared with mild arsenic poisoning group,the ratio of Th17/Treg in severe arsenic poisoning group was elevated (P < 0.05).Correlation analysis showed that the hair arsenic content was positively correlated with the percentage of Th17 cells in peripheral blood lymphocytes and the ratio of Th17/Treg (r =0.323,0.608,P < 0.05),and negatively correlated with the percentage of Treg cells in peripheral blood lymphocytes (r =-0.486,P < 0.05).Conclusion Coal-burning-borne arsenic poisoning can cause the proportion of Th17 cells in the peripheral blood of patients to increase in lymphocytes,and the proportion of Treg cells in lymphocytes to decrease,which in turn changes the balance of Th17/Treg,resulting in weakened immune tolerance and disorder the regulation of inflammatory response,thus participates in the occurrence and development of arsenic-induced immune damage.
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Evaluation of the outcomes of completed projects funded by National Natural Science Foundation of China (NSFC) is a key process of funding management which is of great importance for management improvement and promoting transformation of scientific or technological achievements.Therefore,we evaluated all the projects completed at the end of 2016 funded by NSFC in the field of preventive medicine.The research achievements of selected projects were evaluated,as well as the problems.We hope our work could provide reference and datum for making discipline development strategy in the future.
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Objective To observe the differential expression level of CD4+CD25+Foxp3+regulatory T cells (Treg) in liver of arsenic-exposed rats, explore the regulatory mechanisms on immunological of hepatic injury induced by arsenic, and provide a basis for prevention and treatment of the disease. Methods Thirty-two healthy Wistar rats were selected and randomly divided into control,low,medium and high arsenic dose groups by weight,8 rats per group. Rats in control group were given oral gavage of deionized water, while the other groups were given oral gavage doses of 2.00 g/L sodium arsenite(NaAsO2) according to their body weight for 6 days every week, the concentrations were 1.25, 2.50 and 5.00 ml/kg. After 4 months, liver tissue samples of rats were collected, the content of arsenic in liver was detected by inductively coupled plasma mass spectrometry (ICP-MS);the expression of Treg cells in liver was detected by immunohistochemistry; enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of interloukin-10 (IL-10),transforming growth factor beta 1 (TGF-β1), IL-6, IL-17 and IL-2. Results Compared with the control group [28.57 (17.64 - 35.64)μg/g], the content of arsenic in liver in low,medium and high arsenic exposed groups[M(P25-P75):638.30(527.91-802.58),591.64(513.82-723.16),792.55 (695.93 - 1 074.41) μg/g] increased, the differences were statistically significant(P < 0.05). Compared with low arsenic group, the content of arsenic in liver in high arsenic group increased, the difference was statistically significant (P < 0.05). Numerical density on area (NA) of positive Treg cells in medium,high arsenic exposed groups [(2.25 ± 0.50),(4.00 ± 2.16)A/cm2]was higher than that of the control group[(0.60 ± 0.54)A/cm2,P<0.05];NA of positive Treg cells in high arsenic exposed group was higher than that of the low arsenic exposed group[(1.50 ± 0.58) A/cm2, P < 0.05]. The expressions of the IL-10 in low, medium and high arsenic exposed groups [(5.58 ± 1.70), (6.78 ± 1.09),(7.18 ± 0.53)μg/L]were higher than that of the control group[(2.32 ± 0.83) μg/L,P<0.05];compared with low arsenic group, the expression of IL-10 in high arsenic group increased (P < 0.05); compared with control group [(1.46 ± 0.65) μg/L], the expression of TGF-β1 in high arsenic exposed group increased[(9.06 ± 3.60)μg/L, P<0.05];compared with control group [(2.33 ± 0.66)μg/L], the expression of IL-6 in high arsenic exposed group increased [(5.03 ± 1.39) μg/L, P < 0.05], compared with low arsenic exposed group [(2.46 ± 1.71) μg/L], the expressions of IL-6 in high arsenic exposed group increased, the difference was statistically significant (P < 0.05);the expression of IL-17 among control, low, medium and high arsenic exposed groups[(4.87 ± 1.64),(7.50 ± 2.74), (6.21 ± 1.47),(7.23 ± 2.68)μg/L]were not statistically significant (F = 1.429, P > 0.05); compared with control group [(16.30 ± 3.98) μg/L], the expression of IL-2 in high arsenic exposed group decreased[(9.93 ± 2.65) μg/L, P <0.05]. The content of arsenic in liver was positively correlated with the expression of IL-10, TGF-β1, IL-17, IL-6 (rs=0.696,0.463,0.632,0.502,P<0.05),and negatively correlated with the expression of IL-2(rs=-0.522,P<0.05). Conclusion With increasing of arsenic exposure level, the content of arsenic in liver and the expression of CD4+CD25+Foxp3+Treg have increased,the cytokines are secreted abnormally,liver immunological micro environment is disordered,immune tolerance is formed,and immune clearance is inhibited,which may play an important role in the occur and development of immunological liver damage induced by arsenic in rat.
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Objective To investigate the infiltration of T helper 17 (Th17) and regulatory T cells (Treg) in the liver of rats exposed to arsenic,and to investigate the roles of Th17 and Treg in infiltration of liver injury induced by arsenic.Methods Thirty-two Wistar rats,half male and half female,were randomly divided into control,low,medium and high arsenic dose groups by body weight via the random number table method,8 rats per group.Rats in control group were given oral garage of deionized water,while other groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week,the concentrations of NaAsO2 were 1.25,2.50 and 5.00 ml/kg,respectively.After 4 months,liver tissue samples of rats were collected,the content of arsenic in liver was determined by inductively coupled plasma mass spectrometry (ICP-MS);Hematoxylin-eosin staining (HE) method was used to observe the morphological changes of liver tissue in rats;the protein expressions of interleukin-17A (IL-17A,the imflammatory factor secreted by Th17 cells) and Forkhead Box P3 (Foxp3,the lineage-specific transcription factor of Treg cells) were measured with immunohistochemistry.Results ① Arsenic content in liver of low,medium,and high arsenic exposed groups [63.83 (52.79-80.26),59.16 (51.38-76.58),79.26 (69.59-107.44) μg/g] were higher than those of the control group [2.86 (1.76-3.56)μg/g,P < 0.05],and the high arsenic dose group was higher than the medium arsenic dose group (P < 0.05).② With increasing doses of arsenic,the numbers of inflammatory cells in the liver tissue of rats were increased,and the liver tissue of the high arsenic dose group showed vacuolar degeneration and pathological changes in some areas.③ Compared with the control group,low and medium arsenic dose groups (0.001 + 0.001,0.010 ± 0.020,0.030 ± 0.080),the expression of IL-17A protein in the liver in high arsenic dose group were significantly increased (0.220 ± 0.130,P < 0.05),the differences were statistically significant between groups (F =14.776,P <0.05).The expressions of Foxp3 protein in the liver in low,medium,and high arsenic dose groups were significantly higher (0.270 ± 0.050,0.330 ± 0.040,0.320 ± 0.070) than that in the control group (0.070 ± 0.020),the differences were statistically significant between groups (F =56.990,P < 0.05).④ There was a positive correlation between hepatic arsenic levels and protein levels of IL-17A and Foxp3 in liver (r =0.48,0.81,P < 0.05).Conclusion Arsenic exposure can increase the content of arsenic in liver tissue of rats,which causes the changes of infiltration of Th17 and Treg cells,leading to the change of immune status,suggesting that Thl7 and Treg cells play an important role in the development of arsenic-induced immune injury.
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Objective@#To evaluate of the specificity of a new norovirus (NoV) detection method of in situ capture real-time quantitative reverse transcription polymerase chain reaction (ISC-RT-qPCR) and to apply the method for the detection of NoVs in fresh strawberry.@*Methods@#A panel of stool samples with different NoV genotypes and various inoculums were used for the experiments.@*Results@#We found that all the tested samples of eight genogroup Ⅱ (GⅡ) NoVs could be detected specifically by ISC-RT-qPCR. Moreover, in contrast to the conventional RT-qPCR method , the situation that the Ct value increased as the inoculum of NoV GⅡ decreased was not shown using ISC-RT-qPCR. When we tested NoVs in strawberry samples by ISC-RT-qPCR, the minimum test limit could reach 1.36 genocopy/10 g of fresh strawberry.@*Conclusions@#ISC-RT-qPCR is an effective and specific technic and it could be applied for the detection of infectious NoVs from stool samples and fresh strawberry samples.
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Objective@#To explore the effects of sodium arsenite (NaAsO2) exposure on the activation and extracellular matrix secretion of human hepatic stellate cells, and to provide a theoretical basis for the mechanism study of arsenic induced hepatic fibrosis.@*Methods@#Different doses of NaAsO2 (0.0, 0.1, 1.0, 10.0, 50.0, 100.0 μmol/L) were exposed to human hepatic stellate cell line (Lx-2) for 24, 48 and 72 huors. CCK-8 assay was used to measure cell viability and IC50 of NaAsO2 on Lx-2 was then calculated; According to IC50 results, 0.000, 1.875, 3.750, 7.500, and 15.000 μmol/L of NaAsO2 were exposed to Lx-2 cells for 24 hours, besides, 7.500 μmol/L of NaAsO2 was exposed to Lx-2 cells for 0, 12, 24, 48, and 72 hours, then collected cells and culture supernatant; HSC activation-related protein, including α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) expression levels were detected by Western blot analysis, the main extracellular matrix including laminin (LN) , hyaluronic acid (HA), collagen Ⅳ (COL-Ⅳ) and procollagen Ⅲ(P Ⅲ NP) secretion level was detected by Elisa assay.@*Results@#CCK-8 assay showed that the cell viability of Lx-2 cells were increased obviously at low doses (≤1.0 μmol/L) of arsenic exposure, especially at 48 and 72 h. In contrast, with the increasing doses of arsenic exposure, the survival rate of Lx-2 cell was decreased gradually, and the survival rate of the high-dose (50, 100 μmol/L) arsenic exposure group at 24, 48 and 72 h were significantly lower than 0.0 μmol/L group, P<0.05. The IC50 of NaAsO2 on Lx-2 cells at 24, 48, 72 h were calculated as 72.75, 48.19 and 29.95 μmol/L, respectively; The expression levels of HSC activation-related protein showed that, after treated with 1.875, 3.750, 7.500, 15.000 μmol/L NaAsO2 for 24 h, α-SMA and TGF-β1 protein level were higher than 0.000 μmol/L group. The increased expression of α-SMA and TGF-β1 protein were most significant in 7.500 μmol/L NaAsO2 group (P<0.05). In addition, the expression levels of α-SMA and TGF-β1 also showed a time-dependent increasing in Lx-2 cells after treated with 7.500 μmol/L NaAsO2 for 0, 12, 24, 48 and 72 h; Elisa assay showed that after treated with 1.875, 3.750, 7.500, 15.000 μmol/L NaAsO2 for 24 h, the secretion levels of HA, LN, COL-Ⅳ and PⅢNP were obvious higher than 0.000 μmol/L group (P<0.05). Moreover, the secretion levels of HA, LN, COL-Ⅳ and P Ⅲ NP also showed a time-dependent increased manner in Lx-2 cells after exposed to 7.500 μmol/L NaAsO2 for 0, 12, 24, 48 and 72 h (P<0.05).@*Conclusion@#NaAsO2 exposure to Lx-2 cells can upregulate the expression level of HSC activation-related proteins, induce its further activation, then increase ECM secretion level.