Role of HBV DNA polymerase in mediating the immune escape of tumor cells in HBV-related hepatocellular carcinoma / 临床肝胆病杂志

ObjectiveTo determine whether HBV DNA polymerase is associated with T-cell failure and thus mediates the immune escape of HBV-related hepatocellular carcinoma （HCC） tumor cells， and to investigate the specific molecular mechanisms. MethodsLiver cancer cell lines Huh7 and HepG2 stably transfected with HBV DNA polymerase expression plasmid with Flag （Flag-HBV-P） and intercellular adhesion molecule-1 （ICAM1） were co-cultured with Jurkat cells， and MTT assay， qRT-PCR， and ELISA were used to measure Jurkat cell proliferation， activation （CD69 expression）， and secretion of the cytokine IFN-γ. RNA-seq was used to screen for differentially expressed immune-associated molecules between stably transfected cell lines and control cells， and mRNA half-life and protein half-life assays were used to determine the specific levels of the immune-associated molecules that were affected by HBV DNA polymerase. Related websites were used to predict the transcription factors that may bind to the promoter region of this immune-associated molecule， Western blot was used to verify the effect of transcription factors on the immune-associated molecule， and rescue experiment was used to determine whether HBV DNA polymerase affects the expression level of the immune-associated molecule through this transcription factor. The independent-samples t test was used for comparison between two groups. ResultsThe experimental group had significant reductions in Jurkat cell proliferation， activation， and cytokine secretion compared with the control group （all P<0.01）. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had significant reductions in the mRNA and protein expression levels of ICAM1 （all P<0.01）. Website prediction identified the ICAM1 promoter and preliminarily highlighted NFKB1， RELA， and STAT3. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had a significant reduction in the protein expression level of p65 （all P<0.01）. After p65 overexpression， there was a significant increase in the protein expression level of ICAM1， and after the expression of p65 was reduced， there was a significant reduction in the protein expression level of ICAM1 （all P<0.01）. In the rescue experiment， there was no significant difference in the protein expression level of ICAM1 between the control group and the experimental group after p65 overexpression （all P>0.05）. After the overexpression of ICAM1， there were no significant differences in the proliferation， activation， and cytokine secretion of Jurkat cells between the control group and the experimental group （Huh7 and HepG2 cell lines） （all P>0.05）. ConclusionHBV DNA polymerase downregulates the level of ICAM1 to mediate HCC immune escape by inhibiting the expression of p65 in NF-κB.

Perillyl alcohol modulates activation, permeability and integrity of human brain endothelial cells induced by Plasmodium falciparum

BACKGROUND Cerebral malaria (CM) is a severe immunovasculopathy caused for Plasmodium falciparum infection, which is characterised by the sequestration of parasitised red blood cells (pRBCs) in brain microvessels. Previous studies have shown that some terpenes, such as perillyl alcohol (POH), exhibit a marked efficacy in preventing cerebrovascular inflammation, breakdown of the brain-blood barrier (BBB) and brain leucocyte accumulation in experimental CM models. OBJECTIVE To analyse the effects of POH on the endothelium using human brain endothelial cell (HBEC) monolayers co-cultured with pRBCs. METHODOLOGY The loss of tight junction proteins (TJPs) and features of endothelial activation, such as ICAM-1 and VCAM-1 expression were evaluated by quantitative immunofluorescence. Microvesicle (MV) release by HBEC upon stimulation by P. falciparum was evaluated by flow cytometry. Finally, the capacity of POH to revert P. falciparum-induced HBEC monolayer permeability was examined by monitoring trans-endothelial electrical resistance (TEER). FINDINGS POH significantly prevented pRBCs-induced endothelial adhesion molecule (ICAM-1, VCAM-1) upregulation and MV release by HBEC, improved their trans-endothelial resistance, and restored their distribution of TJPs such as VE-cadherin, Occludin, and JAM-A. CONCLUSIONS POH is a potent monoterpene that is efficient in preventing P. falciparum-pRBCs-induced changes in HBEC, namely their activation, increased permeability and alterations of integrity, all parameters of relevance to CM pathogenesis.

Early and late interventional embolization can improve neurological function, mental state and living ability of patients with intracranial aneurysm / 中国基层医药

Objective:To investigate the application value of early and late interventional embolization in intracranial aneurysms.Methods:Eighty-two patients with intracranial aneurysm who received treatment in Wenzhou People's Hospital from October 2015 to February 2020 were included in this study. These patients were divided into early (≤ 3 days) and late (> 3 days) groups, with 41 patients in each group, according to time from disease onset to surgery. The early group was subjected to early interventional embolization, and the late group was treated with late interventional embolization. The effects of embolization and National Institutes of Health Stroke Scale score pre- and post-treatment, as well as modified Barthel index, Mini-Mental State Exam score, matrix metalloproteinase-9 level, and soluble intercellular adhesion molecule-1 level post-treatment and prognosis were compared between the two groups.Results:The embolization effects in the early group were statistically superior to those in the late group ( P = 0.046). After treatment, National Institutes of Health Stroke Scale score in the early group was significantly lower than that in the late group [(4.02 ± 1.64) points vs. (6.81 ± 2.02) points, t = 6.86, P < 0.01]. Mini-Mental State Exam score and modified Barthel index in the early group were (28.09 ± 1.35) points and (81.12 ± 9.67) points, respectively, which were significantly higher than (26.01 ± 1.19) points and (73.02 ± 8.19) points in the late group ( t = 7.40, 4.09, both P < 0.001). After treatment, matrix metalloproteinase-9 and soluble intercellular adhesion molecule-1 levels in the early group were (420.33 ± 29.40) μg/L and (403.70 ± 23.28) ug/L, respectively, which were significantly lower than (491.30 ± 31.19) μg/mL and (496.37 ± 30.46) μg/L in the late group ( t = 10.60, 15.47, both P < 0.001). Prognosis in the early group was superior to that in the late group ( P = 0.049). Conclusion:Early interventional embolization has better efficacy than late interventional embolization in the treatment of intracranial aneurysm. The former can effectively improve neurological function and mental state, enhance living ability, and improve prognosis, which may be related to the regulation of matrix metalloproteinase-9 and soluble intercellular adhesion molecule-1 levels.

HucMSC-cm protects lipopolysaccharide-induced acute lung injury by activating AMP-activated protein kinase / 中华急诊医学杂志

Objective:To investigate the protective effect of human umbilical cord mesenchymal stem cell conditioned medium (HucMSC-cm) against lipopolysaccharide (LPS)-induced acute lung injury (ALI) and relevant mechanism of action.Methods:Forty 6-week-old male C57BL/6 mice were selected and randomized (random number) into the sham group, LPS group, LPS + HucMSC-cm (LPS+cm) group, and LPS+HucMSC-cm+Compound C (LPS+cm+cc) group, with 10 mice in each group. Mice were intratracheally injected with LPS (5 mg/kg) to establish ALI model, and intratracheally injected with hucMSC-CM (50 μL) 4 h after LPS treatment. Mice in the LPS+cm+cc group were intraperitoneally treated with Compound C (15 mg/kg) prior to LPS treatment. Neutrophils in peripheral blood were counted with the automated hematology analyzer 72 h after LPS administration. After that, mice were sacrificed, and the lung tissue pathology was observed using hematoxylin eosin (HE) staining. Besides, the expressions of IL-6, ICAM-1, VCAM-1 and P-AMP-activated protein kinase (P-AMPK) in the lung tissues were analyzed by Western blot and immunohistochemical assay. In vitro, human lung microvascular endothelial cells (HuLEC-5a) were cultured and divided into three groups: control group, LPS group (10 μg/ mL), and LPS + HucMSC-cm group. After 24 h of treatment, the expressions of p-AMPK and AMPK were detected by Western blot, and the expressions of IL-6 and IL-8 were detected by real-time fluorescence quantitative PCR. Oneway analysis of variance was used to compare the mean values of normally distributed measurement data between groups. Comparisons between two groups were performed using the Tukey’s multiple comparison test. Results:Compared with the sham group, the LPS group showed lungs with congestion and swelling, thickened pulmonary septum, and inflammatory cell infiltration. Moreover, in the LPS group, the protein expressions of IL-6 ( P=0.003), ICAM-1 ( P<0.001) and VCAM-1 ( P=0.001) were increased significantly, while the expression of p-AMPK was decreased ( P=0.013), accompanied by an increase in the proportion of neutrophils in peripheral blood ( P<0.001). Compared with the LPS group, the LPS+HucMSC-cm group demonstrated eased congestion, edema and pathological injury of lung tissue, reversed protein expressions of IL-6 ( P=0.003), ICAM-1 ( P=0.002), VCAM-1 ( P=0.006) and P-AMPK ( P=0.002), as well as decreased proportion of neutrophils in peripheral blood ( P<0.005). Compared with the LPS+HucMSC-cm group, the LPS+cm+cc group exhibited more severe lung histopathological injury, significantly increased protein expressions of IL-6, ICAM-1 and VCAM-1 in lung tissues, as well as decreased expression of P-AMPK protein. The results of immunohistochemistry were consistent with those of protein. In vitro experiment, after LPS treatment, the mRNA expressions of IL-6 ( P<0.001) and IL-8 ( P=0.027) were increased and p-AMPK protein expression ( P=0.005) was decreased as compared with the control group. In comparison with the LPS group, the LPS+HucMSC-cm group showed decreased mRNA expression levels of IL-6 ( P=0.003) and IL-8 ( P=0.002), but increased protein level of p-AMPK ( P=0.003). Conclusions:HucMSC-cm has a protective effect against LPS-induced acute lung injury, which is mainly attributed to the inhibited expression of adhesion molecules and inflammatory factors under the activation of AMPK.

Correlation between adhesion molecules on the surface of red blood cell membrane and red blood cell lifespan / 中国输血杂志

【Objective】 To explore the correlation between red blood cell lifespan and adhesion molecules on the surface of red blood cell membrane, in order to establish a method to detect the duration of red blood cell storage. 【Methods】 10 samples(10 mL each) of fresh red blood cell, collectedf rom 10 healthy voluntary blood donors, were divided into 5 age groups (layers) by Percoll density gradient centrifugation. The expression of CD47, CD44 and CD147 on the surface of red blood cell membrane in each layer was detected using flow cytometry. The variance of protein expression in each layer of red blood cells was analyzed by SPSS statistical software. 【Results】 The expression levels (%) of 3 adhesion molecules on the surface of red blood cell membranes from young to old were CD47: 14.44±2.61, 9.30±1.75, 7.84±1.49, 6.54±1.32 and 5.53±1.12 (P<0.01); CD44: 25.01±1.94, 19.22±1.52, 17.10±1.28, 15.18±1.11 and 13.56±1.08 (P<0.01); CD147: 33.46±1.99, 28.31±2.95, 23.83±1.59, 20.40±1.56 and 18.03±1.65 (P<0.01). 【Conclusion】 The expression levels of CD47, CD44 and CD147 on the surface of red blood cell membranes have showed a downward trend as the storage extended. These three protein adhesion molecules have showed a correlation with red blood cells lifespan, and could be used as detection markers of cell age.

Proliferation and activation of vascular smooth muscle cells induced by activated complement alternative pathway and intervention study / 中国药理学通报

Aim To study the effect of proliferation and activation of vascular smooth muscle cells (VSMCs) induced by activated the complement alternative pathway and intervention. Methods Normal human plasma was specifically activated the complement alternative pathway by incubated with cobra venom factor (CVF). Exposed VSMCs to the activated complement products, the change of cell morphology and the cell viability were assayed by inverted phase contrast microscope and MTT method, respectively. The supernatant was assayed for expression of E-selectin, ICAM-1 and VCAM-1 by using ELISA reagent kits. The protein expression levels of p-NF-kB p65, NF-kB p65 and IKK were assayed by Western blot. The nucleus transcriptional activity of NF-ΚB p65 was tested by the dual luciferase reporter assay system. Pyrrolidine dithiocarbamate (PDTC) was used to intervene the proliferation and activation of VSMCs. Results VSMCs were activated and induced to proliferation after exposed to the products of activated complement alternative pathway. The expressions of E-selectin, VCAM-1 and ICAM-1 were up-regulated. The contents of ICAM-1 and VCAM-1 reached the peak at 6 h and the E-selectin increased significantly at 12 h. Meanwhile, the phosphorylation level of NF-ΚB p65, nucleus transcriptional activity of NF-ΚB p65 and the protein expression of IKK and N F - Κ B p65 increased. The above mentioned changes were clearly inhibited by PDTC. Conclusions The activated complement alternative pathway can initiate proliferation and activation of VSMCs, and its mechanism goes hand in hand with activation of N F - Κ B signaling pathway. PDTC, a specific inhibitor of N F - K B, can effectively inhibit the proliferation and activation of VSMCs.

Expression and significance of L1 cell adhesion molecule and transforming growth factor-β1 in pancreatic cancer tissue / 临床肝胆病杂志

ObjectiveTo investigate the expression of L1 cell adhesion molecule (L1CAM) and transforming growth factor-β1 (TGFβ1) in pancreatic cancer tissue and their association with the prognosis of pancreatic cancer. MethodsHistological specimens were collected from 125 patients with pancreatic cancer who underwent surgical resection in The First Affiliated Hospital of Guangxi Medical University, Guangxi Medical University Cancer Hospital, and Wuming Hospital of Guangxi Medical University from January 2015 to January 2018. Immunohistochemistry was used to measure the expression of L1CAM and TGFβ1 in all specimens, and the association of the expression of L1CAM and TGFβ1 with clinical indices, survival, and prognosis was analyzed. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups; the Cox proportional-hazards regression model was used to investigate the influencing factors for the survival of patients with pancreatic cancer; the Kaplan-Meier survival analysis was used to evaluate the survival of patients with different expression levels of L1CAM and TGFβ1. ResultsThe high protein expression rate of L1CAM in pancreatic cancer tissue was significantly higher than that in adjacent tissue (75.20% vs 20.00%, χ2=76.352, P＜0.001). The high protein expression rate of TGFβ1 in pancreatic cancer tissue was significantly higher than that in adjacent tissue (8160% vs 23.20%, χ2=85.461, P＜0.001). The protein expression of L1CAM was positively correlated with that of TGFβ1 in pancreatic cancer (r=0.492, P＜0.001). The protein expression of L1CAM and TGFβ1 were associated with tumor size, degree of tumor differentiation, TNM stage, lymph node metastasis, intravascular tumor thrombus, and perineural invasion (all P＜0.05). The patients with high protein expression of L1CAM or TGFβ1 had a significantly lower overall survival rate than those with low expression (χ2=54661 and 39597, both P＜0.001). ConclusionL1CAM and TGFβ1 proteins are highly expressed in pancreatic cancer tissue and may be associated with poor prognosis by promoting lymphatic metastasis and hematogenous metastasis. L1CAM and TGFβ1 proteins play an important role in the development, progression, and metastasis of pancreatic cancer.

Comparison of Soybean-based Oil and MCT-olive-fish-soy Oil Intravenous Lipid Emulsions on Soluble Adhesion Markers in Preterm Neonates with Sepsis: A Randomized Controlled Trial

Objectives: To compare the effects of two different intravenous lipid emulsions on solubleadhesion markers in preterm infants with sepsis. Methods: This randomized controlled pilottrial was conducted from February 2016 to February 2017. 40 preterm infants with sepsiswere enrolled and assigned to receive either Medium chain triglyceride-Olive-Fish-Soy lipidemulsion (MOFS-LE) or soybean oil-based lipid emulsion (S-LE). Outcomes of the studywere changes in sICAM-1 and leukocyte integrin β2 levels, and growth after 7 days ofintervention. Results: Leukocyte integrin β2 was significantly higher in MOFS-LE group. Nostatistically significant differences were observed for sICAM-1, duration of mechanicalventilation and antibiotics treatment, and mortality rate. Conclusions: Leukocyte integrin β2was significantly higher in preterm septic neonates who received MOFS-LE

Clinical value of serum cell adhesion molecules in predicting of acute kidney injury in patients with severe sepsis / 中国医师进修杂志

Objective@#To assess the clinical value of serum cell adhesion molecules in predicting of acute kidney injury (AKI) in patients with severe sepsis.@*Methods@#The clinical data of 72 patients with severe sepsis from July 2015 to July 2017 in Sinopharm Dongfeng General Hospital were retrospectively analyzed. Among them, AKI occurred in 45 cases (AKI group), and AKI did not occur in 27 cases (non-AKI group). The clinical data of 2 groups were recorded; the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) on admission were evaluated; the blood routine, serum creatinine, C reactive protein, procalcitonin, lactic acid, intercellular adhesion molecular-1 (ICAM-1), E-selectin, P-selectin and vascular cell adhesion molecule-1 (VCAM-1) were detected.@*Results@#The incidences of infectious shock and respiratory failure, APACHE Ⅱ, SOFA, creatinine, lactic acid, ICAM-1, E-selectin, VCAM-1 in AKI group were significantly higher than those in non-AKI group: 84.4% (38/45) vs. 51.9% (14/27), 51.1% (23/45) vs. 22.2% (6/27), (20.6 ± 2.8) scores vs. (15.9 ± 3.2) scores, (7.4 ± 1.5) scores vs. (4.5 ± 0.7) scores, (131.9 ± 48.7) μmol/L vs. (76.8 ± 10.3)μmol/L, (3.28 ± 0.78) mmol/L vs. (2.64 ± 0.37) mmol/L, (0.93 ± 0.12) mg/L vs. (0.59 ± 0.07) mg/L, (2.25 ± 0.20) mg/L vs. (1.13 ± 0.17) mg/L and (2.65 ± 0.31) mg/L vs. (1.86 ± 0.16) mg/L, and there were statistical differences (P < 0.01 or <0.05); there were no statistical differences in gender, age, basic disease, blood pressure, heart rate, respiratory frequency, infection site, white blood cell, lymphocyte ratio, platelet, blood glucose, C reactive protein, procalcitonin and P-selectin between 2 groups (P>0.05). Pearson correlation analysis result showed that serum ICAM-1, E-selectin, VCAM-1 were positively correlated with serum creatinine (r = 0.625, 0.358 and 0.441; P < 0.05); but there was no correlation between serum lactic acid and serum creatinine (r = 0.146, P > 0.05). Stepwise Logistic regression analysis result showed that creatinine, lactic acid, ICAM-1, E-selectin and VCAM-1 were independent risk factors of AKI in patients with severe sepsis (OR = 2.758, 1.710, 2.955, 1.801 and 2.218; 95% CI 1.845 to 4.371, 1.335 to 2.314, 1.674 to 4.163, 1.572 to 3.235 and 1.339 to 3.752; P < 0.01 or <0.05). The area under curve of serum ICAM-1, E-selectin, VCAM-1 combined detection for predicting of AKI in patients with severe sepsis was 0.876 (95% CI 0.777 to 0.942), with a specificity of 80.0% and a sensitivity of 85.2%.@*Conclusions@#serum ICAM-1, E-selectin, VCAM-1 combined detection can be used as predictor of AKI in patients with severe sepsis.

The relationship between the expression of IL-13 and periostin and the severity of airway remodeling in asthmatic patients / 中国医师杂志

Objective@#By detecting the expression of interleukin-13 (IL-13) and periostin in the airway of asthmatic patients, the pathological changes and pulmonary functions of airway tissues in asthmatic patients were evaluated, and the role of IL-13 and periostin airway remodeling in bronchial asthma was preliminarily explored.@*Methods@#The bronchial tissues adjacent to tumor nest were obtained from 12 patients with lung cancer complicated with bronchial asthma (asthmatic group) and 12 lung cancer patients without bronchial asthma (non-asthmatic group) after lung cancer resection. Pulmonary function was measured for all subjects before surgery. Pathological changes of airway tissues and degree of airway remodeling were assessed by hematoxylin-eosin (H&E) staining, masson′s trichrome staining, and periodic acid-silver methenamine (PASM) staining of paraffin-embedded sections. The expression of IL-13 and periostin in bronchial tissues were evaluated by immunohistochemistry.@*Results@#Values of the forced expiratory volume in 1 second of the predicted value (FEV1% pred) and FEV1/forced vital capacity (FEV1/FVC%) in asthmatic patients were significantly decreased compared with the non-asthmatic patients (P<0.05), indicating that lung function was impaired in asthmatic patients. There was more severe airway remodeling representing as thickening of basement membranes, collagen deposition, and increasing of goblet cells and fibroblasts in asthmatic patients than in non-asthmatic patients (all P<0.05). The expression of IL-13 and periostin were higher in asthmatic tissues than in non-asthmatic tissues (P<0.05). The immunohistochemical expression of IL-13 and periostin in bronchial tissues were positively correlated with the degree of airway remodeling in asthmatic patients, and the expression of IL-13 and periostin in bronchial tissues were positively correlated with each other.@*Conclusions@#The expression of IL-13 and periostin were increased in bronchial tissue in patients with asthma. They work together to promote the occurrence of airway remodeling, which eventually lead to a decline in lung function.

The expression and clinical significance of epithelial cell adhesion molecule and Claudin18 in gastric cancer / 中华老年医学杂志

Objective To investigate the expression and clinical significance of epithelial cell adhesion molecule(EpCAM) and Claudin-18 in gastric cancer.Methods Surgical specimens of gastric cancer were taken from 64 patients.The histological diagnostic criteria were based on WHO standards.Immunohistochemical staining was used to detect expression levels of EpCAM and Claudin-18 in all samples.The relationship between the expression of EpCAM and Claudin-18 and clinicopathological parameters of gastric cancer was analyzed.Correlations of the prognosis of gastric cancer patients with EpCAM and Claudin-18 expression were analyzed using the Cox proportional hazards regression model.Results The positive expression rate of EpCAM was elevated with the increase in tumor size and the occurrence of distant metastasis(x2 =4.526 and 36.090,P=0.033 and 0.000).There were significant differences in Claudin-18 protein expression among different age,TNM stage,growth pattern,depth of invasion and distant metastasis patients(all P＜0.05).Cox regression model analysis showed that tumor size and distant metastasis were the main risk factors affecting the survival of patients with gastric cancer (RR =50.076 and 1.617,P =0.016 and 0.032),while levels of EpCAM and Claudin-18 were not independent risk factors (both P ＞ 0.05).Conclusions EpCAM and Claudin-18 are closely related to the invasiveness of gastric cancer,but abnormally high levels of EpCAM and Claudin-18 are not independent risk factors for the prognosis of gastric cancer patients.

Intervention effect of tetramethylpyrazine on inflammatory response of endothelial cells induced by activated complement alternative pathway / 中国药理学通报

Aim To study the intervention effect of tet- ramethylpyrazine(TMP) on human microvascular endothelial cells (HMECs) inflammatory response induced by activated complement alternative pathway and the possible molecular mechanisms. Methods HMECs were pretreated with different concentrations of TMP, and then exposed to the activated products of the complement alternative pathway which was prepared by cobra venom factor( CVF). The supernatant was removed and assayed for expression of the adhesion molecules (ICAM-1, VCAM-1 and E-selectin) and the inflammatory mediator(IL-6 and TNF-ot) by using ELISA reagent kits. The nucleus transcriptional activity of NF- kB was measured by the dual luciferase reporter assay ? system. Results The adhesion molecules, inflammato ry mediator and nucleus transcriptional activity of NF- kB increased after HMECs were exposed to the products of the activated complement alternative pathway. The up-regulation of ICAM-1, VCAM-1, E-selectin, IL-6, TNF-a and the nucleus transcriptional activity of NF-kB were inhibited by various concentrations of TMP in a dose-dependent manner. Conclusions TMP can effectively reduce inflammatory response of HMECs in-duced by the activated complement alternative pathway , and the mechanism may be highly related to inhibition of nucleus transcriptional activity of NF-kB.

The relationship between the expression of IL-13 and periostin and the severity of airway remodeling in asthmatic patients / 中国医师杂志

Objective By detecting the expression of interleukin-13 (IL-13 and periostin in the airway of asthmatic patients,the pathological changes and pulmonary functions of airway tissues in asthmatic patients were evaluated,and the role of IL-13 and periostin airway remodeling in bronchial asthma was preliminarily explored.Methods The bronchial tissues adjacent to tumor nest were obtained from 12 patients with lung cancer complicated with bronchial asthma (asthmatic group) and 12 lung cancer patients without bronchial asthma (non-asthmatic group) after lung cancer resection.Pulmonary function was measured for all subjects before surgery.Pathological changes of airway tissues and degree of airway remodeling were assessed by hematoxylin-eosin (H&E) staining,masson's trichrome staining,and periodic acid-silver methenamine (PASM) staining of paraffin-embedded sections.The expression of IL-13 and periostin in bronchial tissues were evaluated by immunohistochemistry.Results Values of the forced expiratory volume in 1 second of the predicted value (FEV1％ pred) and FEV1/forced vital capacity (FEV1/FVC％) in asthmatic patients were significantly decreased compared with the non-asthmatic patients (P ＜ 0.05),indicating that lung function was impaired in asthmatic patients.There was more severe airway remodeling representing as thickening of basement membranes,collagen deposition,and increasing of goblet cells and fibroblasts in asthmatic patients than in non-asthmatic patients (all P ＜ 0.05).The expression of IL-13 and periostin were higher in asthmatic tissues than in non-asthmatic tissues (P ＜ 0.05).The immunohistochemical expression of IL-13 and periostin in bronchial tissues were positively correlated with the degree of airway remodeling in asthmatic patients,and the expression of IL-13 and periostin in bronchial tissues were positively correlated with each other.Conclusions The expression of IL-13 and periostin were increased in bronchial tissue in patients with asthma.They work together to promote the occurrence of airway remodeling,which eventually lead to a decline in lung function.

Upregulation of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in renal tissue in severe dengue in humans: Effects on endothelial activation/dysfunction

Abstract INTRODUCTION: Dengue is an important mosquito-borne disease in tropical and subtropical regions. Adhesion molecules have not been systematically characterized in the renal tissue of patients with severe dengue (SD). The objective of this study was to detect viral antigens in samples from patients that evolved with SD, correlating with the expression of ICAM-1, VCAM-1, VE-cadherin, and E-selectin to contribute to a better understanding of the pathophysiology of SD. METHODS: Kidney specimens from patients with SD were selected according to clinical and laboratorial data and submitted to histological and immunohistochemistry analysis. A semiquantitative evaluation was performed considering positive immunostaining in 20 glomeruli. RESULTS: Viral antigens were mainly detected in distal tubules. The intense immunostaining of VCAM-1 and ICAM-1 was observed. The expression of E-selectin was discrete, and VE-cadherin expression varied from mild to moderate. VCAM-1 was slightly intense in the glomerular capsule; the expression of ICAM-1 was diffuse. E-selectin was diffuse, and VE-cadherin varied from mild to moderate. The most frequent histological findings were glomerular congestion, mild glomerulitis, acute renal injury, and glomerular atrophy. CONCLUSIONS: The results appear to demonstrate an imbalance between vascular endothelial permeability regulating events in renal lesions in SD. The increase in the expression of ICAM-1 and VCAM-1 is an in-situ indicator of higher permeability with a consequent influx of cells favoring the inflammation of the endothelium. These molecules are important in the pathophysiology of the disease and provide the possibility of developing new markers for the evaluation, clinical follow-up, and therapeutic response of patients with SD.

Comparison of CD9 & CD146 markers in endometrial stromal cells of fertile & infertile females

Background & objectives: CD9 and CD146 are important adhesion molecules that play a role in the implantation of an embryo. This study was undertaken to correlate the expression of these markers in fertile and infertile women's endometrial stromal cells. Methods: Human endometrial stromal cell culture from endometrial biopsies of fertile (n=50) and infertile females (n=50) was performed and primary cell lines were established. Expression of CD9 and CD146 was studied for all the 100 cell lines with the help of flow cytometry. Gene expression of CD9 and CD146 was performed by real-time polymerase chain reaction. Results: There was a significant difference in endometrial stromal cells of fertile and infertile females. Flow cytometric results revealed significantly lower expression of CD9 (P=0.0126) and CD146 (P=0.0006) in the infertile endometrial stromal cells as compared to fertile endometrial stromal cells. These results were comparable with real-time data. Interpretation & conclusions: This study showed that endometrial stromal cells from infertile females had lower expression of adhesion molecules, CD9 and CD146. Our findings suggest that CD9 and CD146 may have a role in infertility. Infertile female's endometrial stromal cells have decreased expression of CD9 and CD146 which can be the cause of infertility related to implantation failure.

Novel mutations of ITGB2 induced leukocyte adhesion defect type 1 / 中华儿科杂志

Objective@#To investigate the pathogenic mechanism of two novel ITGB2 mutations in leukocyte adhesion defect type 1 (LAD1).@*Methods@#The clinical history and blood sample of an 11 years old patient admitted to Affiliated Hospital of Qingdao University in August 2014 were collected. Expression of CD18 (encoded by ITGB2) was analyzed by flow cytometry. Novel ITGB2 mutations were identified by next-generation sequencing technology and confirmed by Sanger sequencing. The functional effect of ITGB2 mutations was detected by PolyPhen2. Expression vectors of both wild type and mutant ITGB2 were constructed and transfected into mammalian cells for analysis of protein stability and subcellular location.@*Results@#The symptoms of the patient (recurrent infections, lowered alveolar ridge and hypodontia) supported the diagnosis of LAD1. Expression of CD18 on the leukocytes was significantly decreased (0.2%) compared with the control samples from the parents (paternal: 99.0%; maternal: 99.1%). The patient was identified to be compound heterozygous for ITBG2 c.954del G (novel mutation) and c.1802C>A (paternal originated). ITGB2 c.954 del G was confirmed to be a harmful frameshift mutation; ITGB2 c.1802C>A was also predicted to be harmful. In terms of protein stability. There was no significant difference between mutant D18 and wild type. However, subcellular location analysis showed the mutant D18 could not locate on cell membrane.@*Conclusion@#The compound heterozygous of ITGB2 mutations (c.954del G and c.1802C>A) decreases the expression and impairs the location of CD18 on leukocytes, which leads to LAD1.

Expression of adhesion molecules on CD34+ cells from steady-state bone marrow before and after mobilization and their association with the yield of CD34+ cells

BACKGROUND: Cell adhesion molecules (CAMs) expressed on hematopoietic progenitor cells (HPCs), endothelial cells, and stromal cells play a pivotal role in the mobilization of CD34+ cells. Herein, we conducted a non-randomized peripheral blood stem cell (PBSC) mobilization study aimed to compare the potential differences in the expressions of several CAMs and chemokines on CD34+ cells obtained from bone marrow aspirate before and after HPC mobilization from patients with hematologic malignancies and healthy donors. METHODS: Three-color cytofluorometric analysis was used to compare the expressions of CAMs and chemokines in the bone marrow before and after mobilization. RESULTS: For all studied groups, CAM expression among those with good and poor yields of CD34+ cells was significantly correlated with VCAM-1 (P=0.007), CD44 (P=0.027), and VLA-4 (P=0.014) expressions. VCAM-1 (P=0.001), FLT-3 (P=0.001), CD44 (P=0.011), VLA-4 (P=0.001), and LFA-1 (P=0.001) expressions were higher before HPC mobilization than after HPC mobilization. By contrast, the expression of CXCR4 significantly varied before and after mobilization only among those with successful PBSC mobilization (P=0.002). CONCLUSION: We attempted to identify particular aspects of CAMs involved in CD34+ cell mobilization, which is a highly complex mechanism that involves adhesion molecules and matrix metalloproteases. The mechanism by which CD34+ cell mobilization is activated through proteolytic enzymes is not fully understood. We believe that CXCR4, VLA-4, CD44, and VCAM-1 are the most important molecules implicated in HPC mobilization, particularly because they show a correlation with the yield of CD34+ cells collected via large volume leukapheresis.

Expression of tumor stem cell marker EpCAM in patients undergoing liver transplantation for hepatocellular carcinoma and its relationship with tumor recurrence and metastasis / 中华普通外科杂志

Objective To investigate the expression of EpCAM (epithelial cell adhesion molecule) in patients undergoing liver transplantation for hepatocellular carcinoma (HCC),and to explore the relationship between EpCAM expression level and HCC recurrence.Methods 83 HCC tissue samples were collected and analyzed retrospectively.EpCAM was detected by immunohistochemistry staining and the correlation between EpCAM and clinicopathological features,prognosis and recurrence were analyzed retrospectively from patients undergoing liver transplantation in Peking University People's Hospital from 2000 to 2010.Results Log-rank univariate survival analysis showed that the 5-year overall survival (41.2％ vs.73.3％,x2 =4.935,P =0.026) and 5-year disease free survival (41.2％ vs.73.3％,x2 =4.634,P =0.031)of HCC patients with high EpCAM expression level was significantly lower than that with low expression level of EpCAM.COX multivariate survival analysis showed patients with high EpCAM expression had a higher risk of recurrence(HR =2.860,95％ CI:1.012-8.083) and death (HR =2.909,95％ CI:1.030-8.217)after liver transplantation than those with low EpCAM expression,which was an independent predictor of 5-year overall survival and 5-year disease free survival recurrence (P =0.044).Furthermore,EpCAM expression level was highly related to tumor distant metastasis (P =0.01).Conclusion There was positive relation between high expression of EpCAM and high HCC recurrence after liver transplantation,suggesting that EpCAM can be a predictor for HCC recurrence and long-term survival of patients with HCC after transplantation.

Expression of E-cadherin, Slug and NCAM and its relationship to tumor invasiveness in patients with acromegaly

Pituitary adenomas account for 10-15% of primary intracranial tumors. Growth hormone (GH)-secreting adenomas account for 13% of all pituitary adenomas and cause acromegaly. These tumors can be aggressive, invade surrounding structures and are highly recurrent. The objective of this study was to evaluate E-cadherin, Slug and neural cell adhesion molecule (NCAM) expression in GH-secreting pituitary adenomas and its relationship to tumor invasiveness. A cross-sectional study of patients who underwent hypophysectomy due to GH-secreting pituitary adenoma from April 2007 to December 2014 was carried out. The medical records were reviewed to collect clinical data. Immediately after surgery, tumor samples were frozen in liquid nitrogen and stored in a biofreezer at -80°C for assessment of E-cadherin 1 (CDH1), SLUG (SNAI2), and NCAM (NCAM1) by real-time PCR. The samples were fixed in formalin and embedded in paraffin for immunohistochemical analysis of E-cadherin and NCAM. Thirty-five patients with acromegaly were included in the study. Of these, 65.7% had invasive tumors. Immunohistochemically, E-cadherin was expressed in 96.7% of patients, and NCAM in 80% of patients. There was no statistically significant relationship between tumor grade or invasiveness and immunohistochemical expression of these markers. Regarding gene expression, 50% of cases expressed CDH1, none expressed SNAI2, and 53.3% expressed NCAM1. There was no statistically significant relationship between tumor grade or invasiveness and gene expression of CDH1, SNAI2, and NCAM1. The absence of Slug overexpression and of E-cadherin and NCAM suppression suggests that expression of these markers is not associated with tumor invasiveness in GH-secreting pituitary adenomas.

Effects of Alcohol Extracts of Bark and Male Flower of Eucommia ulmoides Oliv. on Airway Allergic Inflammation of Model Mice / 中国中医药信息杂志

Objective To investigate the effects of alcohol extract of bark and male flower of Eucommia ulmoides Oliv. on airway allergic inflammation induced by chicken ovalbumin (OVA) in mice; To explore its mechanism of action. Methods On day 0, day 7, mice were intraperitoneally injected OVA for sensitization, followed by nasal stimulation for 21 days to establish airway allergic inflammation mice models. The mice were divided into normal group, model group, alcohol extract of bark of Eucommia ulmoides Oliv. group, alcohol extract of male flower of Eucommia ulmoides Oliv.group,and Dexamethasone group.Each medication group was given relevant medicine for gavage. The lung tissue was embedded in HE and PAS dyeing, to observe the pathological changes of bronchus and surrounding lung. The levels of serum OVA-IgE, IL-4, IFN-γ and IL-13 were measured by ELISA. The expression of ICAM-1, VEGF, MMP9 and TIMP1 were detected by immunohistochemistry. Flow cytometry was used to detect the expression of Th17 cells in peripheral blood. The expressions of TNF-α and IL-6 mRNA in lung tissue were detected by RT-PCR. Results The model group showed changes of airway allergic inflammatory such as eosinophils and other inflammatory cell infiltration, bronchial spasm, and mucus secretion. Lung histopathology in alcohol extract of bark and male flower of Eucommia ulmoides Oliv.groups was improved significantly(P<0.05).Compared with the normal group, the levels of serum OVA-IgE, IL-4 and IL-13 increased in model group, while the level of IFN-γ decreased (P<0.05, P<0.01). The expressions of ICAM-1, VEGF and MMP9 increased, while the expression of TIMP1 decreased (P<0.01); peripheral blood IL-17+cells increased (P<0.01); the expressions of TNF-α and IL-6 mRNA increased. Compared with the model group, the levels of serum OVA-IgE, IL-4 and IL-13 decreased in alcohol extract of bark and male flower of Eucommia ulmoides Oliv. groups (P<0.05, P<0.01); the expressions of ICAM-1 and VEGF decreased (P<0.05, P<0.01); the expression of TIMP1 increased. Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.could down-regulate IL-17+cells,reduce the expression of IL-6 mRNA(P<0.05,P<0.01). Alcohol extract of bark of Eucommia ulmoides Oliv. group could induce the secretion of IFN-γ (P<0.01), and down-regulate the expression of TNF-α mRNA(P<0.05).Alcohol extract of male flower of Eucommia ulmoides Oliv. group could significantly down-regulate the expression of MMP9 (P<0.05). Conclusion Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.can induce the production of OVA-IgE,inhibit secretion of Th2 cytokines, inhibit the expression of adhesion molecules, depress Th17 cells, so as to inhibit the airway allergic inflammation.