Expression of programmed death-1 and programmed death-ligand 1 in post-transplant lymphoproliferative disorders after allogeneic hematopoietic stem cell transplantation in children / 中华实用儿科临床杂志

Objective:To explore the expressions of programmed death-1 (PD-1) and programmed death-ligand 1 (PD-L1) and clinicopathological characteristics in post-transplant lymphoproliferative disorder (PTLD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children, with the aim of clarifying whether checkpoint inhibition of PD-1/PD-L1 inhibitors may serve as a therapy option.Methods:The clinical data of 13 cases of PTLD after allo-HSCT pathologically confirmed in Shenzhen Children′s Hospital from January 1, 2012 to December 30, 2019 were retrospectively analyzed.The detection was performed by immunohistochemical staining by MaxVision? method, Epstein-Barr virus(EBV) in situ hybridization and lymphoma gene rearrangement.The relationship between the expression of PD-1 and PD-L1 and the clinicopathological characteristics of PTLD were analyzed.Results:The expression of PD-1 was not correlated with gender, age, primary diseases, histopathological types, transplantation mode and the expression of EBV in situ hybridization (all P>0.05). The expression of PD-L1 was correlated with histopathological types ( P<0.05). Furthermore, the expression rate of PD-L1 on severe β-thalassemia was significantly higher than that of severe aplastic anemia [90.0%(9/10 cases) vs. 66.7%(2/3 cases)] and monomorphic PTLD was higher than that of polymorphic PTLD [100.0%(2/2 cases) vs. 83.3%(5/6 cases)]. Moreover, the positive PTLD in EBV was higher than the negative PTLD in EBV [90.9%(10/11 cases) vs. 50.0%(1/2 cases)]. The positive rates of PD-1 and PD-L1 in 13 cases with PTLD were 46.2%(6/13 cases) and 61.5%(8/13 cases) in tumor cells, 92.3% (12/13 cases) and 76.9% (10/13 cases) in microenvironmental cells, and 84.6%(11/13 cases) in EBV, respectively. Conclusions:PD-L1 has a higher positive rate in tumor cells with monomorphic PTLD; and routine staining for PD-1 and PD-L1 can be performed in all types of PTLD when standard immunotherapy and chemotherapy are ineffective.

Application of the mortality risk score for severe hand, foot and mouth disease in children with hand, foot and mouth disease in pediatric intensive care unit / 中国小儿急救医学

Objective@#To analyze the predictive value of the mortality risk score for severe hand, foot and mouth disease(MRSHFMD) system for the complications and mortality risk of severe hand, foot and mouth disease(HFMD) in children.@*Methods@#This study included 354 children with severe HFMD who admitted in the pediatric intensive care unit(PICU) of Hunan Provincial Children′s Hospital from March 2012 to March 2014.The patients were grouped according to whether they had complicated nervous system damage, pulmonary edema, pulmonary hemorrhage and circulatory failure in the course of disease, and the prognosis was grouped according to their 28 d survival.The worst values of white blood cell count, blood glucose, blood lactic acid, N-terminal pro-brain natriuretic peptide, within 24 hours after admission were used to score MRSHFMD.The predictive value of MRSHFMD for nervous system damage, pulmonary edema, pulmonary hemorrhage, circulatory failure, and prognosis were evaluated using the receiver operating characteristic(ROC)curve.@*Results@#The blood glucose, white blood cell count, blood lactic acid value, N-terminal pro-brain natriuretic peptide and MRSHFMD score of the children with HFMD complicated with nervous system damage, pulmonary edema, pulmonary hemorrhage and circulatory failure were significantly higher than those in the non-complicated groups(P<0.01). When the cut-off value of MRSHFMD score was 3, the area (95%CI) under the ROC curve were 0.723 (0.643-0.804), 0.870 (0.793-0.946), 0.921 (0.85-0.992), 0.944 (0.867-1.000) and 0.954 (0.000-1.000) of nervous system damage, pulmonary edema, pulmonary hemorrhage, circulation failure and death in children with HFMD, respectively.The specificity and sensitivity of predicting nervous system damage, pulmonary edema, pulmonary hemorrhage, circulatory failure and death were 44.6% and 95.8%; 67.5% and 95.5%; 83.3% and 95.1%; 89.3% and 95.1%; 90.9% and 93.7%, respectively.@*Conclusion@#MRSHFMD system is an effective tool to predict HFMD complications of pulmonary hemorrhage, circulatory failure, and death, which is worthy of clinical promotion.

The clinical value of cerebrospinal fluid tumor necrosis factor-α and resolvin D1 in neonatal purulent meningitis / 中国新生儿科杂志

Objective To study the clinical value of tumor necrosis factor-α (TNF-α) and resolvin D1 (RvD1) concentrations in cerebrospinal fluid (CSF) of neonatal purulent meningitis(NPM).Method From June 2016 to June 2017,neonates of suspected NPM admitted to the neonatology department of our hospital were studied prospectively.Their CSF was examined before the use of antibiotics.The patients were assigned into NPM group and non-NPM group.After 7 to 10 days of treatment,according to the clinical symptoms and the reexamination results of CSF,patients in the NPM group were further assigned into the improved group and the unimproved group.The levels of TNF-α and RvD1 in CSF were measured using enzyme-linked immunosorbent assay (ELISA) method,and SPSS 22.0 was used for statistical analysis.Result A total of 23 patients were included in the NPM group (18 in the improved group and 5 in the unimproved group) and 30 in the non-NPM group.The levels of TNF-α and RvD1 in the CSF of the NPM group were higher than the non-NPM group [TNF-α:(0.263 ±0.088) pg/ml vs.(0.087 ±0.001) pg/ml,RvD1:(2.017 ± 0.171) pg/ml vs.(0.563 ±0.048) pg/ml] (P ＜0.05).After 7 to 10 days of treatment,TNF-α and RvD1 decreased in the improved NPM group[TNF-α:0.083 (0.078,0.111) pg/ml vs.0.122 (0.098,0.214) pg/ml,RvD1:1.242 (0.740,2.098) pg/ml vs.1.791 (1.371,2.804) pg/ml] (P ＜ 0.05),and increased in the unimproved NPM group [TNF-α:2.239 (1.309,2.806) pg/ml vs.0.102 (0.100,1.312) pg/ml,RvD1:2.614 (1.265,2.940) pg/ml vs.0.139 (0.103,0.276) pg/ml] (P ＜ 0.05).The reexamination results of TNF-oα in the NPM group were lower than the examination results before the use of antibiotics of the non-NPM group,and RvD1 higher than the non-NPM group (P ＜ 0.05).Conclusion TNF-α and RvD1 in CSF have clinical value for the early diagnosis and therapeutic evaluation of NPM.

Application of the mortality risk score for severe hand, foot and mouth disease in children with hand, foot and mouth disease in pediatric intensive care unit / 中国小儿急救医学

Objective:To analyze the predictive value of the mortality risk score for severe hand, foot and mouth disease(MRSHFMD) system for the complications and mortality risk of severe hand, foot and mouth disease(HFMD) in children.Methods:This study included 354 children with severe HFMD who admitted in the pediatric intensive care unit(PICU) of Hunan Provincial Children′s Hospital from March 2012 to March 2014.The patients were grouped according to whether they had complicated nervous system damage, pulmonary edema, pulmonary hemorrhage and circulatory failure in the course of disease, and the prognosis was grouped according to their 28 d survival.The worst values of white blood cell count, blood glucose, blood lactic acid, N-terminal pro-brain natriuretic peptide, within 24 hours after admission were used to score MRSHFMD.The predictive value of MRSHFMD for nervous system damage, pulmonary edema, pulmonary hemorrhage, circulatory failure, and prognosis were evaluated using the receiver operating characteristic(ROC)curve.Results:The blood glucose, white blood cell count, blood lactic acid value, N-terminal pro-brain natriuretic peptide and MRSHFMD score of the children with HFMD complicated with nervous system damage, pulmonary edema, pulmonary hemorrhage and circulatory failure were significantly higher than those in the non-complicated groups( P<0.01). When the cut-off value of MRSHFMD score was 3, the area (95% CI) under the ROC curve were 0.723 (0.643-0.804), 0.870 (0.793-0.946), 0.921 (0.85-0.992), 0.944 (0.867-1.000) and 0.954 (0.000-1.000) of nervous system damage, pulmonary edema, pulmonary hemorrhage, circulation failure and death in children with HFMD, respectively.The specificity and sensitivity of predicting nervous system damage, pulmonary edema, pulmonary hemorrhage, circulatory failure and death were 44.6% and 95.8%; 67.5% and 95.5%; 83.3% and 95.1%; 89.3% and 95.1%; 90.9% and 93.7%, respectively. Conclusion:MRSHFMD system is an effective tool to predict HFMD complications of pulmonary hemorrhage, circulatory failure, and death, which is worthy of clinical promotion.

Effects of miR-200c-3p on proliferation and apoptosis of nephroblastoma SK-NEP-1 cells / 中华实用儿科临床杂志

Objective:To investigate the effect of miR-200c-3p on the proliferation and apoptosis of nephroblastoma SK-NEP-1 cell and its mechanism.Methods:From January 2015 to August 2019, nephroblastoma tissue and peritumoral tissue of 30 patients in Shenzhen Children′s hospital were collected.The experimental group of mimic negative control, miR-200c-3p and miR-200c-3p inhibitor was set up.The expression of miR-200c-3p in 30 paired nephroblastoma tissues and adjacent kidney tissues were detected by real-time fluorescence quantitative PCR.The differential expression of miR-200c-3p was also detected in SK-NEP-1 and 293FT cell lines.The effects of miR-200c and miR-200c-3p inhibitor on the proliferation, cell cycle distribution and apoptosis of SK-NEP-1 cells were detected by cell counting kit-8(CCK-8)and flow cytometry assays, respectively.Xenograft tumors were generated by peri-renal adipose tissue injection to assess the effect of miR-200c-3p on tumor growth in vivo.The pathological morphology of xenograft tumors was observed by HE staining.The proliferation index of Ki-67 were detected by immunohistochemistry.Western blot method was used to detect the B-cell lymphoma 2(Bcl-2), Bcl-2 related X protein (Bax) and cleaved cysteinyl aspartate specific proteinase-3(Caspase-3)expression level on xenograft tumors. Results:The expression level of miR-200c-3p in nephroblastoma(0.420±0.587)was significantly lower than that in matched normal kidney(1.500±0.504)( t=8.613, P<0.001). The expression level of miR-200c-3p in SK-NEP-1 cells (0.363±0.006) was significantly lower than that in human embryonic kidney 293FT cells (0.807±0.186) ( t=4.136, P<0.05). The group and time interaction results of CCK-8 proved that miR-200c-3p could inhibit the proliferation of SK-NEP-1 cells( F=16.81, P<0.001). The flow cytometer test of cell cycle and apoptosis showed that miR-200c-3p could block in G0/G1 and S phase( t=-7.770, P<0.01; t=11.501, P<0.001). Moreover, it increased the early apoptosis rate and decreased the late apoptosis rate ( t=-22.270, P<0.001; t=4.612, P<0.01). A orthotopic transplantation assay was employed to evaluate the effect of miR-200c-3p and miR-200c-3p inhibitor on the proliferation of SK-NEP-1 cells.The final volume of tumor miR-200c-3p group [(0.419±0.16) cm 3]was significantly lower than those in the control group [(2.469±0.914) cm 3, t=0.507, P<0.001]. However, the miR-200c-3p inhibitor group had no significant difference [(1.627±0.189) cm 3; t=2.209, P=0.052]. miR-200c-3p overexpression upregulated expression levels of apoptotic proteins cleaved Caspase-3 and Bax ( t=-47.000, -82.730, all P<0.001), but downregulated the expression level of anti-apoptotic protein Bcl-2( t=53.740, P<0.001). Conclusions:The overexpression of miR-200c-3p can inhibit the proliferation, promote the apoptosis of the SK-NEP-1 cells and partially inhibited tumorigenicity of nude mouse acted as a tumor suppressor gene.

MicroRNA-200c-3p inhibits proliferation of nephroblastoma cells by targeting CCNE2 / 南方医科大学学报

OBJECTIVE@#To predict and verify the target gene of miR-200c-3p and evaluate the inhibitory effect of miR-200c-3p on the proliferation of nephroblastoma cells.@*METHODS@#The putative target genes of miR-200c-3p were predicted by bioinformatics approach. Nephroblastoma cell models with miR-200c-3p overexpression or knockdown were established in SK-NEP-1 and G401 cells with corresponding control groups. The expressions of CCNE2 in SK-NEP-1 and G401 cells in different groups were detected by RT-PCR and Western blotting. A luciferase reporter assay was used to determine the targeting relationship between miR-200c-3p and CCNE2. The effects of miR-200c-3p overexpression or knockdown on cell proliferation was detected by cell counting kit-8 (CCK-8) assay and soft agarose assay.@*RESULTS@#CCNE2 was one of the target genes of miR-200c-3p as predicted by bioinformatics methods. Transfection of the two nephroblastoma cell lines with miR-200c-3p mimic resulted in significantly lowered CCNE2 mRNA and protein expressions ( < 0.05). The results of dual-luciferase assay confirmed that miR-200c-3p bound to the 3'UTR of CCNE2. CCK-8 assay and soft agarose assay demonstrated that overexpression of miR-200c-3p significantly inhibited the proliferation of the nephroblastoma cells ( < 0.01), and knocking down miR-200c-3p in the cells produced the opposite effects.@*CONCLUSIONS@#miR-200c-3p overexpression inhibits the proliferation of nephroblastoma cells by down-regulating its target gene CCNE2.

Research progress of tyrosine kinase inhibitors in the treatment of pulmonary fibrosis / 中国综合临床

Objective Pulmonary fibrosis is a progressive chronic lung disease with a high incidence.Although the path of the disease has not been fully elucidated,the pathogenesis of the disease is roughly similar.Tyrosine kinases are involved in a series of signaling pathways that are critical for cell homeostasis.Substantial evidence from in vitro studies and experimental animal models suggests that tyrosine kinases play a role in promoting the development and progression of pulmonary fibrosis,and tyrosine kinase inhibitors have shown good anti-fibrosis and anti-inflammatory effect in animal models of pulmonary fibrosis.

Effect of nilotinib on silicon dioxide-induced proliferation and collagen synthesis in HFL-1 cells / 中国职业医学

OBJECTIVE: To observe the effects of nilotinib on silicon dioxide(SiO_2)-induced cell proliferation and collagen synthesis in human fetal lung fibroblast-1(HFL-1) cells and to explore the related mechanism. METHODS: ⅰ) HFL-1 cells were induced with different doses of SiO_2 suspension(0, 5,10, 25, 50 and 100 mg/L) for 24.0 hours. The expression of transforming growth factor-β1(TGF-β1), C-Abl, and platelet-derived growth factor receptor(PDGFR) was detected by Western blot, and the dose of SiO_2 in subsequent experiments was screened. ⅱ) HFL-1 cells were randomly divided into 6 groups: 1) the control group: no treatment; 2) the solvent control group: cells were treated with 0.10% dimethyl sulfoxide; 3) the SiO_2 stimulation group: cells were induced with SiO_2 suspension at a dose of 50 mg/L for 24.0 hours; 4)-6) the nilotinib groups: cells were induced with SiO_2 suspension at a dose of 50 mg/L for 24.0 hours and treated with nilotinib at the concentration of 5, 10, or 15 mmol/L for 24.0 hours. Cell proliferation was detected by MTS assay. The TGF-β1 protein secreted by cells was measured using enzyme linked immunosorbent assay. The expression of TGF-β1, C-Abl, platelet derived growth factor(PDGF), PDGFR and collagen typeⅠproteins was measured by Western blot. RESULTS: ⅰ) The dose of the SiO_2 in the experiments was set to 50 mg/L. ⅱ) The cell proliferation rate of HFL-1 cells in the SiO_2 stimulation group and the 3 nilotinib groups was higher than that in control group and solvent control group(P<0.05). The proliferation rates of HFL-1 cells in 10 and 15 mmol/L nilotinib groups were lower than that in SiO_2 stimulation group(P<0.05). The level of TGF-β1 and the protein relative expression levels of TGF-β1, collagen typeⅠ, C-Abl, PDGFR and PDGF in HFL-1 cells of SiO_2 stimulation group were higher than those in control group and solvent control group(P<0.05). The above indexes of HFL-1 cells in 15 mmol/L nilotinib group were lower than that in SiO_2 stimulation group(P<0.05); the above indexes of HFL-1 cells in 5 mmol/L nilotinib group were not significantly different from those in SiO_2 stimulation group(P>0.05). The level of TGF-β1 and the relative expression level of C-Abl protein in HFL-1 cells of 10 mmol/L nilotinib group were lower than those in SiO_2 stimulation group(P<0.05). CONCLUSION: Nilotinib can inhibit the proliferation of HFL-1 cells and reduce the expression of collagen typeⅠprotein induced by SiO_2. This process may be achieved by inhibiting tyrosine kinase-mediated signaling pathway.

Effect of sorafenib on growth and circadian clock gene expression of human hepatocellular carcinoma in nude mice / 中华肝胆外科杂志

Objective To investigate the function of sorafenib on the growth of hepatocellular carcinoma by establishing subcutaneous transplantation tumor model with nude mice.To explore the effect of sorafenib on circadian clock gene expression in hepatoma cells.Methods Mouse tumor model was established by implanting hepatocarcinoma cell (HepG2) subcutaneously in Balb/C nude mice.Sixteen experimental mice were randomly divided into two groups:sorafenib treatment group (n =8) and solvent control group (n =8).The nude mice were treated with sorafenib (100 mg/kg) and DMSO daily by intragastric administration,respectively.The volume of tumors was recorded every 3 days.The expressions level of circadian clock genes (Per1,Per2,Per3,CLOCK,Cry1,Cry2,BMAL1 and CKIε) were detected by real-time polymerase chain reaction (Real-time PCR).The correlations between the size of the xenografts and the expressions of the circadian clock genes were further analyzed.Results Compared with the control group,the tumor size in the sorafenib treatment group were significantly smaller comparing with the control group.Results of Real-time PCR showed that the expression level of Per1,Cry1 and BMAL1 mRNA was remarkably decreased in the treatment group (Per1,P =0.02;Cry1,P =0.002;BMAL1,P =0.035),the differences were statistically significant.Correlation analysis showed that the size of subcutaneous transplantation tumorsin nude mice was negatively correlated with the expressions of Per1,Per2,Cry1 and Cry2 mRNA in control group.While,the size of subcutaneous transplantation tumors was negatively correlated with the expressions of Per2,Per3 and BMAL1 levels in the sorafenib treatment group.Conclusions There is a negative correlation between the expression levels of some biological clock genes and the size of transplantation tumor in nude mice.Sorafenib treatment significantly inhibited the growth of hepatocellular carcinoma in nude mice and down-regulation the expressions of Per1,Cry1 and BMAL1 mRNA in hepatoma cells.

Fibroblast growth factor 10 inhibits lipopolysaccharide-induced microglial activation / 中国病理生理杂志

AIM: To investigate the effects of fibroblast growth factor 10 ( FGF10 ) on lipopolysaccharide ( LPS)-induced microglial activation .METHODS:Mouse BV2 microglial cells were maintained in DMEM in a humidified incubator with 95%/5%( V/V) mixture of air and CO 2 at 37℃.The medium was changed every 1 or 2 d.The cells were digested and passaged every 4 or 5 d.The BV2 microglial cells were first pretreated with FGF 10 (1 mg/L) for 30 min and then stimulated with LPS (500 μg/L).The medium and the cells were collected at different time points .The morphologi-cal changes of microglia were visualized under microscope .To evaluate the microglial activation , the transcription and pro-duction of proinflammatory factor tumor necrosis factor-α( TNF-α) were examined by real-time quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively.RESULTS:The morphology of control BV2 microglia showed circular or oval shape .After exposure to LPS for 24 h, the microglia revealed spindle shaped or multipolar morphology , and the percentage of activated cells was significantly increased compared with control group.Pretreatment with FGF10 successfully inhibited the morphological change from normal to activated shape .LPS sti-mulation for 6 h significantly increased the transcription of TNF-α, while FGF10 pretreatment remarkably reversed the effect.In addition, the production of TNF-αincreased in the presence of LPS stimulation for 24 h compared with control group.Pretreatment with FGF10 suppressed LPS-induced TNF-αexpression.CONCLUSION: Pretreatment with FGF10 inhibits the morphological change from normal to activated shape , and remarkably suppressed the transcription and produc-tion of TNF-α.FGF10 successfully suppresses LPS-induced BV2 microglial activation , indicating that FGF10 is a therapeu-tic agent for the treatment of glia-mediated neuroinflammatory diseases .

Application Progress of Exonuclease-assisted Signal Amplification Strategies in Biochemical Analysis / 分析化学

As an important member of tool enzymes, exonuclease is a kind of hydrolytic enzymes without strict base sequence dependent. In recent years, by taking advantage of different hydrolysis ways of exonuclease and nanotechnology, cycle effect of enzyme digestion, aptamer, non Watson-Crick base pairing system by metal ions, fluorescent nucleic acid probes, electrochemical methods etc. , a series of exonuclease-assisted signal amplification strategies have been developed, which played a very key role in improving the sensitivity of detection methods. Therefore, exonuclease has been widely used in high sensitive detection of nucleic acids, proteins, ions, small molecules and so on. To understand it better and apply it well in the future, the application progress of exonuclease-assisted signal amplification strategies in biochemical analysis has been summarized in this review.

A TiO2/CNTs Nanocomposites Enhanced Luminol Electrochemiluminescence Assay for Glucose Detection / 分析化学

Abstract A novel luminol electrochemiluminescence strategy based on titanium dioxide/carbon nanotubes ( TiO2/CNTs) nanocomposites for detection of glucose was developed. First, the TiO2/CNTs nanocomposites were prepared by a sol-gel method and modified on the glassy carbon electrode. The electrochemiluminescence ( ECL) signal could be greatly enhanced when the electrode was established by the nanocomposites, which finally resulted in the increased sensitivity. Glucose oxidase calalyzed the oxidation of glucose to form H2 O2 , and the H2 O2 reacted with luminol to produce the ECL signal. Thus the above system was proved to be efficient for glucose detection. The modified electrode exhibited excellent ECL signals and a good linear range of 1. 0í10-7-5. 0í10-6 mol/L with a detection limit of 5. 2í10-8 mol/L towards glucose detection. This strategy was successfully demonstrated as a sensitive, rapid, simple and cost-effective method to detect glucose. Meanwhile, the TiO2/CNTs nanocomposites offered a novel material for the signal enhancement in electrochemiluminescence sensor.

Induced transformation of rat oval cells line WB-F344 and tumor formation in NOD/SCID mice / 临床与实验病理学杂志

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

Three-dimensional case teaching method and its application to anesthesia teaching / 中华医学教育探索杂志

There are many subjects related to anesthesiology and the band between anesthesiology and clinical practicevery is close. So students can not be proficient in anesthesiology by traditional teaching methods. The use of three-dimensional case teaching method in anaesthesiology teaching is a good attemptment. three-dimensional case teaching method integrate case method, multimedia technology and clinical case. It can help medical students analyze and solve clinical problems, improvetheir academic performance and enhance their clinical basic skills.