The evaluation value of heparin-binding protein levels in bronchoalveolar lavage fluid for severe pneumonia in children / 中国小儿急救医学

Objective:To investigate the value of the level of heparin-binding protein(HBP)in bronchoalveolar lavage fluid(BALF)on the evaluation of severe pneumonia in children.Methods:A total of 94 children with severe pneumonia who underwent bronchoscopy and bronchoalveolar lavage were admitted at Hunan Children′s Hospital, and HBP levels in BALF were detected.According to the etiological results, the patients were divided into non-bacterial infection group(19 cases) and bacterial infection group(75 cases). According to the existence and severity of acute respiratory distress syndrome (ARDS), the cases were divided into non-ARDS group(65 cases), mild ARDS group(23 cases) and moderate to severe ARDS group(6 cases).Results:The HBP level of BALF in the bacterial infection group was higher than that in the non-bacterial infection group, and the difference was statistically significant[ 20.77(5.90, 73.50)ng/mL vs.5.9(5.90, 7.64)ng/mL, Z=12.500, P<0.001]. The HBP level of BALF in the moderate to severe ARDS group[300.00(169.29, 300.00)ng/mL] was significantly higher than those in the non-ARDS group[11.90(5.90, 36.95)ng/mL] and the mild ARDS group[15.13(7.41, 46.44)ng/mL], and the difference was statistically significant( H=14.718, P=0.001). In predicting the presence of bacterial infection in severe pneumonia, the area under the receiver operating characteristic curves of BALF HBP, serum procalcitonin (PCT) and serum C-reactive protein(CRP) were 0.758, 0.737, and 0.732, respectively.When the optimal truncation values of BALF HBP, serum PCT and serum CRP were 8.40 ng/mL, 0.16 ng/mL, and 8.39 mg/L, the predicted sensitivities were 70.7%, 69.3%, 46.7%, and the predicted specificity were 79.0%, 79.0%, 94.7%, respectively. Conclusion:The level of HBP in BALF in children with severe pneumonia increases with the severity of ARDS, and significantly increases in the positive group of bacterial infection, which can be used as one of the auxiliary indicators to evaluate the severity of severe pneumonia and bacterial infection in children.

The evaluation value of heparin-binding protein in pulmonary infection and severity of pulmonary infection / 中国小儿急救医学

Severe pulmonary infection is one of important causes of death for children under five years old in China, and early identification of the cause and severity of lung infections is important for guiding clinical interventions. Heparin-binding protein (HBP) is a protein with bactericidal activity, and its concentration is very low in healthy people. Lung bacterial infection, acute lung injury or acute respiratory distress syndrome, which can lead to permeability of vascular endothelial cells increased, thus HBP in blood or tissue fluid can be significantly increased. It has certain guiding significance for the early identification of bacterial infection and judgment of the severity of the disease. This study reviewed the evaluation role of HBP in lung infection and severity of illness.

Mechanism of MALAT1 induced osimertinib resistance in HCC827 lung cancer cells / 中华肿瘤杂志

Objective@#To test the effect of metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) and/or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib.@*Methods@#We transfected HCC827 cells with LV-vector or LV-over/MALAT1. Stable transfected cells (HCC827/Vector, HCC827/MALAT1) were selected by adding puromycin. HCC827/MALAT1 cells were further transfected with the shRNA-negative control (NC) or shRNA-human epidermal growth factor receptor 3 (ERBB3) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib on the proliferation of HCC827 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and/or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib treatment were detected by western blot.@*Results@#The MTT assay showed that sensitivity to osimertinib of HCC827/MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50) of osimertinib >4 000 nmol/L in HCC827/MALAT1 cells. However, knockdown of ERBB3 facilitated the anti-proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA-ERBB3 cells was (17.27±3.21) nmol/L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827/MALAT1 cells induced by 10 nmol/L osimertinib was (8.38±0.92)%, significantly lower than (27.17±5.83)% of knockdown of ERBB3 (P<0.01). Western blotting showed that the expression of p-ERBB3, p-AKT and p-extracellular regulated protein kinases (ERK) in HCC827/MALAT1 cells was markedly up-regulated, while the expression of p-epithelial growth factor receptor (EGFR) was inhibited. The expressions of p-ERBB3, p-AKT and p-ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p-EGFR, p-ERBB3, p-AKT and p-ERK in ERBB3 deleted cells.@*Conclusions@#MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3/PI3K/AKT and ERBB3/MAPK/ERK signaling pathways.

Expression of PLOD2 in esophageal squamous cell carcinoma and its correlation with invasion and metastasis / 中华病理学杂志

Objective@#To investigate PLOD2 expression in esophageal squamous cell carcinoma, and to explore the potential mechanism by which PLOD2 promotes tumor metastasis.@*Methods@#The expression of PLOD2 in 60 cases of esophageal squamous cell carcinoma (the patients were collected at the first Affiliated Hospital of Xinxiang Medical University, from January 2016 to December 2017) was investigated by immunohistochemistry. Fibrillar collagen formation and collagen deposition were detected by picrosirius red staining. Correlation of PLOD2 expression with clinical pathologic features of the patients was performed using χ2 test and Kaplan-Meier analysis. After EC-109 cells were transfected with LV-vector and LV-over/PLOD2, the expression of PLOD2 was detected by real time PCR and the impact of POLD2 on invasion in EC-109 cells was determined by transwell migration and invasion assays. The expression of PLOD2/AKT epithelial-to-mesenchymal transition signal pathway related proteins was detected by Western blot.@*Results@#The expression level of PLOD2 in esophageal squamous cell carcinoma was 81.7% (49/60 cases),higher than their paired noncancerous tissues(8.3%, 5/60; P<0.01), and correlated significantly with tumor depth of invasion and nodal metastasis (P<0.01). Picrosirius red staining showed that collagen deposition was increased and the degree of fibrillar organization was enhanced in carcinoma tissues that had higher PLOD2 expression. Transwell migration and invasion assays showed that PLOD2 significantly promoted the migration and invasion ability of EC-109 cells. Western blot showed that PLOD2 significantly increased the expression levels of p-FAK, p-AKT and vimentin in EC-109 cells.@*Conclusions@#Esophageal squamous cell carcinoma has a high expression of PLOD2 that correlates with tumor invasion and lymph node metastasis. PLOD2 promotes invasion and metastasis of esophageal squamous cell carcinoma through epithelial-to-mesenchymal transition via FAK/AKT signal pathway.

Upregulation of PLOD2 promotes invasion and metastasis of osteosarcoma cells / 中华肿瘤杂志

Objective@#To investigate the relationship of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2.@*Methods@#The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT-PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan-Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U-2OS cells with LV-vector, LV-over/PLOD2, sh-NC and sh-PLOD2. The expression of PLOD2 was detected by qRT-PCR. The impact of POLD2 on U-2OS cell invasion was determined by wound-healing assay and Transwell migration assay. The expressions of PLOD2/FAK/JAK2-STAT3 signal pathway related proteins were detected by western blotting.@*Results@#The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues (P<0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P<0.01). The same results were also observed in qRT-PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 (P<0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+ 9.63)%, significantly higher than (9.67±1.28)% in tissue with low expression of PLOD2 (P<0.001). The result of wound-healing and Transwell migration assay showed that over-expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U-2OS cells (both P<0.01). The result of western blotting showed that over-expression of PLOD2 significantly increased the expression levels of p-FAK, p-JAK2, p-STAT3, but knockdown PLOD2 decreased the levels of p-FAK, p-JAK2, p-STAT3 in U-2OS cells.@*Conclusions@#Up-regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK/JAK2-STAT3 signal pathway.

A systematic review of the mechanisms and influence factors of cancer-related fatigue / 中国医师杂志

Cancer-related fatigue (CRF) is one of the most common symptoms in clinical cancer patients.It is associated with cancer itself or with cancer-related therapies.It is a persistent and subjective tiredness experience.It not only affects the patient;s physical,psychological and physical condition,social function,quality of life,but also may cause interruption of cancer-related treatment and affect the patient's life.However,the etiology and mechanism of CRF are not fully understood.Many foreign scholars believe that the occurrence of CRF may be related to anemia,obesity,tumor type,insomnia,inflammatory cytokines,hypothalamic-pituitary-adrenal axis and so on.As cancer patients demand higher quality of life,CRF are increasingly receiving medical attention.In this paper,the related factors and mechanisms of CRF are stated.

Mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1 induced invasion and metastasis of esophageal cancer cell EC-109 / 中华肿瘤杂志

Objective@#To investigate the effect and mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1, (LncRNA-MALAT1) on invasion and metastasis of esophageal cancer cell EC-109.@*Methods@#EC-109 cells were transfected with lentiviral vector carrying short hairpin RNA of MALAT1( shRNA-MALAT1) or a nonspecific shRNA control (shRNA-control). The expressions of MALAT1, microRNA-200a, ZEB1 and ZEB2 were detected by qRT-PCR. The effect of shRNA-MALAT1 on invasion of EC-109 cells was determined by transwell assay. The expressions of components of epithelial-msenchymal transition pathway in EC-109 cells were determined by immunofluorescence array and western blotting. The expression relationship between MALAT1 and miR-200a in EC-109 cells was detected by dual-luciferase reporter assay.@*Results@#The result of qRT-PCR showed that the expressions levels of MALAT1, ZEB1 and ZEB2 in shRNA-MALAT1 group were 0.43±0.06, 0.64±0.04 and 0.51±0.04, respectively, significantly lower than 0.97±0.08, 1.06±0.07 and 0.98±0.05 in shRNA-control group and 1 in control group, respectively(all P<0.05). Transwell assay showed that the number of invaded cells in shRNA MALAT1 group was (96.81±10.43) per low-power field, markedly lower than that of (278.44±13.28) per low-power field in shRNA-control group (P<0.01). Immunofluorescence staining and Western blotting showed that MALAT1 downregulation significantly reduced the expressions of proteins related to EMT signal pathway in EC-109 cells.Dual luciferase reporter assay showed that compared to negative control, the activities of luciferase reporter in EC-109 cells co-transfected with pmirGLO-MALAT1-wt and miR-200a were significantly down-regulated. While co-transfected pmirGLO-MALAT1-mut with miR-200a mimics had no effect on the luciferase reporter activities of MALAT1.@*Conclusion@#LncRNA MALAT1 functions as a competing endogenous RNA to regulate the expressions of ZEB1 and ZEB2 by sponging miR-200a and promotes invasion and migration of esophageal cancer cells through inducing epithelial-mesenchymal transition.

Effect of Bufalin on apoptosis in human esophageal cancer through inhibiting NF-κB / 实用医学杂志

Objective To observe the effect of Bufalin on controlling ECA109 cells , and to explore its potential anti-tumor mechanism. Method The effect of Bufalin on the proliferation of human esophageal cancer ECA109 cells was evaluated by CCK-8 assay and it effects on apoptosis of esophageal cancer ECA109 cells were determined by flow cytometry (FCM). Protein expressions of NF-κBp65, ERK, PERK1/2 and Caspase-3, Cleaved Caspase-3, PARP, Cleaved PARP in esophageal cancer ECA109 cell were observed through Western blot. Results The proliferation of human esophageal cancer ECA109 cells was significantly inhibited in bufalin group in a time- and concentration-dependent manner. Bufalin can induce the apoptosis in human esophageal ECA109 cells. Results of Western blot showed the protein expressions of apoptosis-related protein Cleaved Caspas-3 and Cleaved PARP in esophageal cancer ECA109 cells could be markedly up-regulated by Bufalin , but p-ERK1/2, NF-κBp65 in esophageal cancer ECA109 cells could be markedly down-regulated by Bufalin. Conclusion Bufalin can potently inhibit the growth of esophageal cancer ECA109 cells and the potential anti-tumor mechanism might be involved in inhibiting ERK/ NF-κB signal pathway.

Bufalin reverses hepatocyte growth factor-induced resistance to afatinib in H1975 lung cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of bufalin in reversing hepatocyte growth factor (HGF)-induced resistance to afatinib in H1975 lung cancer cells, and explore its possible mechanism.</p><p><b>METHODS</b>The afatinib-resistant H1975 lung cancer cells (H1975AR) were induced by exogenous HGF and transfected with recombinant adenoviral vector Ad-HGF-GFP. The cytostatic effects of bufalin, afatinib and bufalin plus afatinib on H1975AR cells were evaluated by MTT assay. The impact of combined therapy with bufalin and afatinib on invasion of H1975AR cells was determined by transwell migration assay. The concentrations of HGF in the culture supernatants of H1975/Vec and H1975/HGF cells were determined by ELISA. The expression of EGFR, cMET and EMT signal pathway-related proteins in H1975AR cells treated with bufalin, afatinib and bufalin plus afatinib were detected by Western blot.</p><p><b>RESULTS</b>The results of MTT assay showed that afatinib did not inhibit the growth of H1975 cells, but after 72 h of the combined treatment with bufalin and afatinib and in the presence of HGF, the growth rate of H1975 cells was (38.67 ± 8.76)%, significantly lower than the growth rate of (63.45 ± 12.65)% in the H1975 cells treated with HGF alone (P < 0.05). The results of transwell migration assay showed that in the presence of HGF, afatinib plus bufalin combination therapy markedly decreased the number of invaded H1975 cells through the Matrigel chamber (48.98 ± 11.43), significantly lower than the 118.92 ± 37.29 of afatinib-treated or the 88.84 ± 19.53 of bufalin-treated cells (P < 0.05 for all). The result of ELISA showed that H1975/HGF cells secreted high levels of HGF, and afatinib and bufalin had no effect on the HGF secretion in H1975/HGF cells. The results of Western blot analysis showed that the expression of p-EGFR, p-cMet, p-AKT, p-ERK, vimentin and snail in H1975AR cells treated with bufalin puls afatinb was down-regulated markedly, and the expression of E-cadherin was up-regulated markedly.</p><p><b>CONCLUSIONS</b>Combination of bufalin and afatinib strongly inhibits the growth of H1975AR lung cancer cells and decreases their invasion ability. The possible mechanism of combined treatment with bufalin and afatinib may be related to the blocking of cMet/PI3K/AKT and cMet/MAPK/ERK pathways and inhibiting of epithelial-mesenchymal transition.</p>