Objective To investigate the effects of emulsified isoflurane on apoptosis in human neuroblastoma cells and the role of c-Jun N-terminal kinase (JNK) in it.Methods The human neuroblastoma SHSY-5Y cells were seeded in 96-well plates or dishes and then randomly divided into 8 groups using a random number table:control group(group C,n=24),different concentrations of lipid emulsion groups(LE1 groups [n =24],LE2 group [n =24] and LE3 group [n =72]),different concentrations of emulsified isoflurane groups (EI1 group [n=24],El2 group [n=24] and EI3 group [n=72]),and emulsified isoflurane + JNK inhibitor SP600125 group (group EI-SP,n =24).At 24 h after the cells were plated,in LE1-3 groups,30％ lipid emulsion was added to the culture medium with the final concentrations of 0.395 6,0.791 2 and 1.582 4 μl/ml,respectively;in EI1-3 groups,8％ emulsified isoflurane was added with the final concentrations of 0.56,1.12 and 2.24 mmol/L,respectively;in group EI-SP,emulsified isoflurane was added with the final concentration of 1.12 mmol/L,and SP600125 was added at 1 h before addition of emulsified isoflurane with the final concentration of 10 μmol/L;the cells were cultured normally in group C.At 6,12 and 24 h of incubation in EI3 and LE3 groups,and at 24 h of incubation or culture in the other groups,the morphology of cells was detected,the cell viability was measured using methyl thiazolyl tetrazolium assay,and the expression of JNK,phosphorylated JNK (p-JNK) and cytochrome c (Cyt c) was detected by Western blot.Results Compared with group C,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in group EI2 and at 12 and 24 h of incubation in group EI3,the cell viability was significantly decreased (P＜0.05),and no significant change was found in the expression of p-JNK and Cyt c in group EI-SP,and no significant change was found in the cell viability and expression of p-JNK and Cyt c in LE1-3 and EI1 groups (P＞0.05).Compared with group EI1,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in EI2.3 groups (P＜0.05).Compared with group EI2,the cell viability was significantly decreased,and the expression of p-JNK and Cyt c was significantly up-regulated at 24 h of incubation in group EI3,and the cell viability was significantly increased,and the expression of p-JNK and Cyt c was significantly down-regulated in group EI-SP (P＜0.05).There was no significant difference in JNK expression between the eight groups (P＞0.05).Conclusion High concentrations of emulsified isoflurane can induce apoptosis in neurons only when applied for a long time,while low concentrations do not have the effect when applied for a short time.The mechanism by which emulsified isoflurane induces neuronal apoptosis is related to activation of JNK pathway.

OBJECTIVETo investigate the significance of beta-adrenergic receptor 2 (beta2-AR) and vascular endothelial growth factor-2 (VEGFR-2) in the occurrence and development of infantile hemangioma through detecting the expression of beta2-AR and VEGFR-2 in the different stages of infantile hemangiomas.

METHODSAccording to the Mulliken's classification standard, we classified the specimens as proliferating group (32 cases), involuting group (17 cases) and involuted group (11 cases). Normal skin tissue surrounding the hemangioma from 7 cases were chosen as control group. The expression of beta2-AR and VEGFR-2 was detected by immunohistochemical technique in proliferating hemangioma, involuting hemangioma, involuted hemangioma. The mean optical density was measured by image analysis system (Image Pro Plus 6.0) and SPSS 16.0 software was applied for statistical analysis.

RESULTSThe expression of beta2-AR and VEGFR-2 was strongly positive in proliferating hemangioma, while positive in involuting hemangioma and weakly positive in the involuted stage. The mean optical density of each phase was 0.064 751 2 +/- 0.012 747, 0.031 6017 +/- 0.006 848,0.011 869 8 +/- 0.039 349 for beta2-AR, and 0.068 940 9 +/- 0.029 274, 0.028 445 5 +/- 0.006 396, 0.011 184 1 +/- 0.004 198 for VEGFR-2. The differences between different stages had a statistically significance (P < 0.05). Correlation analysis on the mean optical density between beta2-AR and VEGFR-2 had a statistically significance (P < 0. 05).

CONCLUSIONSBeta2-AR and VEGFR-2 may be involved in the occurrence and development of infantile hemangioma.

Child ; Child, Preschool ; Female ; Hemangioma ; metabolism ; Humans ; Infant ; Male ; Receptors, Adrenergic, beta-2 ; metabolism ; Receptors, Vascular Endothelial Growth Factor ; metabolism

Objective To investigate the mechanisms underlying the effect of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on cutaneous squamous cell carcinoma (SCC) in mice.Methods A model of cutaneous SCC was established in 21 SKH-1 hairless mice,which were treated with topical ALA 8％ cream followed by single irradiation with He-Ne laser at a total dose of 30 J/cm2 (ALA-PDT).Three mice were sacrificed before and at 1,3,6,12,24 hours and 7 days after the irradiation,separately,and SCC tissue was taken from the mice.Transmission electron microscopy (TEM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were performed to determine the pattern of tumor cell death(necrosis,apoptosis and autophagy) during 1-24 hours after ALA-PDT,and immunohistochemical techniques were used to estimate the expressions of LC3B and CD34 on SCC cells,as well as the quantity of CD1a+ cells,CD4+ T and CD8+ T lymphocytes in SCC tissue 7 days after the irradiation.Statistical analysis was done by two-sample t test using SPSS 17.0 software.Results TEM showed gradual necrosis and apoptosis (especially necrosis) of tumor cells and formation of autophagosomes in macrophages within 24 hours after ALA-PDT.The number of apoptotic cells per high power field (× 400) in SCC tissue significantly increased at 24 hours compared with that before ALA-PDT (7.30 ± 2.18 vs.2.00 ± 0.69,P ＜ 0.05).As immunohistochemistry revealed,there was a significant decrease in the number of CD34+ cells (1.33 ± 0.58 vs.19.00 ± 2.66,P＜ 0.01),but a marked increase in that of CD1a+ ce1ls (23.01 ± 2.04 vs.10.33 ± 1.88,P＜ 0.05),CD4+ T cells (28.67 ± 1.76 vs.12.40 ± 2.27,P＜ 0.05),CD8+ T cells (25.79 ± 2.37 vs.11.67 ± 1.45,P ＜ 0.05) and LC3B+ interstitial cells (30.6 ± 3.21 vs.21.44 ± 4.3,P ＜ 0.05) per high power field (× 400) in SCC tissue on day 7 compared with that before ALA-PDT.Conclusions ALA-PDT may directly kill SCC cells by inducing cell necrosis and apoptosis rather than autophagy.Additionally,ALA-PDT can injure microvascular endothelial cells and cause the aggregation of dendritic cells,CD4+ T cells and CD8+ T cells in SCC tissue.

Objective To investigate the clinical efficacy of conventional treatment combined with flupentixol and melitracen in patients with reflux esophagitis (RE).Methods From June 2012 to March 2015, a total of 182 patients were selected as study subjects from newly diagnosed RE patients.The anxiety and depression scores were evaluated according to Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD).And then patients were divided into HAMA and HAMD negative conventional treatment group and combined treatment group, HAMA and HAMD positive conventional treatment group and combined treatment group.Rabeprazole and mosapride were administrated in conventional treatment group.For patients in combined treatment group, on the base of conventional treatment flupentixol and melitracen were added.The treatment course was eight weeks.The degree of anxiety and depression, RE symptoms and mucosal healing under gastroscope were evaluated before and after treatment.Adverse drug reaction was observed.Chi square test or t test was performed for statistical analysis.Results Eight weeks after treatment, the scores of HAMA and HAMD in HAMA and HAMD positive combined treatment group were 7.930 ±3.832 and 9.630 ± 3.650, which were both lower than those of conventional treatment group (11.660 ± 4.108 and 12.170 ± 4.459), and the differences were statistically significant (t=3.683 and 2.233;both P＜0.05).The symptom scores of heartburn, regurgitation, chest pain of HAMA and HAMD positive combined treatment group were 0.700±0.591,0.780± 0.629 and 0.720±0.621, respectively, which were lower than those of conventional treatment group (1.280 ± 0.502, 1.370 ± 0.610 and 1.040 ± 0.842), and the differences were statistically significant (t =5.133, 4.413 and 2.114, all P＜0.05).There were no statistical significance in symptoms scores between HAMA and HAMD negative combined treatment group and conventional treatment group (all P＞0.05).After treatment, the mucosal healing rate of HAMA and HAMD positive combined treatment group was 91.3％ (42/46), which was higher than that of conventional treatment group (71.7 ％, 33/46), and the difference was statistically significant (x2 =5.845, P =0.016).The incidence of adverse events of HAMA and HAMD negative combined treatment group was 4.8％ (2/42), and that of HAMA and HAMD positive combined treatment group was 2.2％(1/46).Conclusions The conventional treatment combined with flupentixol and melitracen in RE patients accompanied with anxiety and depression was remarkable and safe.RE patients without obvious anxiety or depression, preventive use of antianxiety and antidepressant medicine can not improve the efficacy.

Objective To analyze the differences of the clinical and neuropathological features among the common Charcot-Marie-Tooth disease (CMT) subtypes.Methods There were 81 CMT patients confirmed by genetic testing from 2005 to 2015 in Department of Neurology,Peking University First Hospital,including 31 cases of CMT1A (38.3％),19 cases of CMTX1 (23.5％),16 cases of CMT2A2 (19.8％) and 15 cases of 9 rare types of CMT (1.2％-4.9％).We compared the onset age,duration,muscles weakness of legs,frequency of pes cavus,and main pathological changes of the sural nerve biopsy in 48 cases of the common CMT subtypes.Results The mean age of the onset was (12.00 ± 6.77) years in CMT1A patients,(11.81 ±4.65) years in CMTX1 patients and (5.00 ±2.68) years in CMT2A2 patients (Brown-Forsythe test,P =0.001).The duration was (12.00 ± 6.75) years in CMT1A patients,(8.50 ± 4.75) years in CMTX1 patients and (5.00 ± 2.73) years in CMT2A2 patients (Brown-Forsythe test,P =0.001).The muscle force of the dorsi flexors was Ⅳ (0,Ⅴ) in CMT1A patients,Ⅲ + (0,Ⅳ) in CMTX1 patients and 0 (0,Ⅳ) in CMT2A2 patients (H =11.359,P =0.020).The pes cavus appeared in 15/23 cases of CMT1A,10/16 cases of CMTX1 and 1/9 cases of CMT2A2 (Fisher test,P=0.017).The leukoencephalopathy appeared only in 3 cases of CMTX1 and the visual loss appeared only in 3 cases of CMT2A2.The onion-bulb formations of myelinated fibers appeared in 23/23 cases of CMT1 A,5/16 cases of CMTX1 and 2/9 cases of CMT2A2(Fisher test,P =0.000).The axonal regeneration appeared in 16/23 cases of CMT1A,16/16 cases of CMTX1 and 9/9 cases of CMT2A2 (x2 =7.666,P =0.016).There were significant differences among the three common CMT subtypes in the above parameters.Conclusions CMT1A,CMT2A2 and CMTX1 are the most common subtypes of CMT in the present study.For the clinical diagnosis,more attention should be paid to the onset of the disease,duration,muscles weakness,pes cavus,cerebral symptoms and visual loss.The present frequency of onion-bulb and the axonal regeneration of myelinated fibers help the different pathological diagnosis among them.

Objective To assess the effectiveness and safety of propranolol in infantile hemangiomas by comparing with prednisone.Methods A systematic literature search of PubMed,Embase,Cochrane,Ovid,Google Scholar and CNKI,VIP,Wanfang database was conducted to identify studies about the treatment of propranolol in children with hemangiomas.We chose randomized controlled trials and clinical controlled trials.We selected literatures by certain standards.Results Eight papers including 9 studies were identified by the strategy mentioned above.These 8 literatures met our inclusion criteria after review by two independent reviewers.The studies comprised 407 patients.Six of the control group were oral prednisone,and there was no statistic heterogeneity (P =0.09,I2 =0％).The fixed model was used to do the statistic analysis.The outcome showed the effective rate of propranolol was higher than that of prednisone,with statistically significant difference (OR=7.56,95％ CI:3.18-17.98).Three of the control group included observation or oral placebo,without statistic heterogeneity (P=0.48,I2=0％) either.The outcome showed the effective rate of propranolol on hemangioma was higher than that of the control group (OR=23.15,95％CI:7.15-74.94).Of all the eight researches,five reported adverse effects,with statistic heterogeneity (P=0.0003,I2 =81 ％).In addition,the adverse rate of propranolol was lower than that of prednisone,with statistically significant difference (OR=0.12,95％ CI:0.02-0.75).Conclusions The results of this meta-analysis show that oral propranolol could obviously decrease the volume and improve the color of infantile hemangiomas.And propranolol is a significantly more effective for IH than steroids.The incidence of adverse effects of propranolol is also lower than that of prednisone.As a result,propranolol should be recommended as the first choice therapy for infantile hemangioma.

Objective:To investigate the application value of spectral computed tomo-graphy (CT) multi-parameter imaging in predicting gastric cancer lymph node metastasis.Methods:The retrospective case-control study was conducted. The clinicopathological data of 86 patients with gastric cancer who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to June 2017 were collected. There were 53 males and 33 females, aged from 22 to 87 years, with a median age of 53 years. All patients received abdominal plain scan and arterial and venous phase contrast spectral scan. Images of plain scan, 70 keV monochromatic and iodine-based images in arterial and venous phase were analyzed on post-processing working station. Observation indicators: (1) gastric cancer lymph node metastasis; (2) analysis of influencing factors for lymph node metastasis in gastric cancer; (3) introduction of special cases. Count data were described as absolute numbers. Univariate analysis was performed using the chi-square test or rank sum test. Multivariate analysis was conducted using the Logistic regression model.Results:(1) Gastric cancer lymph node metastasis: of the 86 patients, 64 cases had lymph nodes metastasis and 22 had no lymph nodes metastasis. (2) Analysis of influencing factors for lymph node metastasis in gastric cancer: results of univariate analysis showed that tumor growth pattern, tumor diameter, infiltration of peritumor fat, CT value in arterial phase, CT value in venous phase, iodine value in venous phase were related factors affecting lymph nodes metastasis in gastric cancer ( χ2=6.753, Z=-3.180, χ2=7.649, Z=-2.051, -2.971, -2.547, P<0.05). Results of multivariate analysis showed that infiltration of peritumor fat and the iodine value in venous phase were greater than 12(100 μg/cm 3) and not greater than 16(100 μg/cm 3), or greater than 16(100 μg/cm 3) were independent risk factors affecting lymph nodes metastasis in gastric cancer ( odds ratio=13.154, 3.761, 7.583, 95% confidence interval as 2.597-66.620, 1.893-8.572, 4.769-16.692, P<0.05). (3) Introduction of special cases: case 1 was male, aged 46 years. Results of preoperative spectral CT enhanced scan showed gastric antrum space occupying lesion combined with enlarged lymph nodes. During enhancement arterial phase, spectral CT 70 keV monochromatic images and corresponding iodine-based images of primary lesion layer in the transverse view showed gastric wall thickening with mild to moderate enhancement, clear fat space in serosa and enlarged lymph nodes in lesser curvature. The spectral CT 70 keV monochromatic images and corresponding iodine-based images below pylorus level in the transverse view showed subpyloric enlarged lymph nodes. During enhancement venous phase, the 70 keV monochromatic images and corresponding iodine-based images of primary lesion layer in the transverse view showed layered enhancement of gastric antrum lesions and mucosal enhancement, with a high iodine value. The patient was diagnosed as gastric antrum cancer with lymph node metastasis, no serosal or peritumoral fat invasion. Results of postoperative pathological examination showed moderately differentiated adenocarcinoma of gastric antrum with serosal invasion and lymph node metastasis. Case 2 was male, aged 53 years. Results of preoperative spectral CT enhanced scan showed gastric cancer of lesser curvature combined with enlarged lymph nodes. During enhancement arterial phase, 70 keV monochromatic images and corresponding iodine-based images of primary lesion layer in the transverse view showed gastric wall heterogeneous thickening of lesser curvature, with moderate enhancement, obscure peritumor fat space, unclear serosa, and multiple enlarged lymph nodes in lesser curvature. During enhancement venous phase, 70 keV monochromatic images in the transverse view showed unclear boundary between lesions and enlarged lymph nodes in lesser curvature, obscure peritumor fat. During enhancement arterial phase, 70 keV monochromatic images of celiac trunk layer in the transverse view showed parasplenic artery lymph nodes, with circular enhancement and no enhancement in central necrotic elements. The patient was diagnosed as gastric cancer of lesser curvature with lymph node metastasis, serosal and peritumor fat invasion. Results of postoperative pathological examination showed poorly differentiated adenocarcinoma of gastric antrum with serosal invasion and lymph node metastasis. Conclusion:The infiltration of peritumor fat and iodine value in venous phase are independent factors affecting gastric cancer lymph node metastasis.

Objective To evaluate the effect of dexmedetomidine administered at different time points on the quality of anesthesia recovery and inflammatory response in children undergoing craniotomy.Methods According to the randomized double-blind method,200 pediatric patients who underwent craniotomy in Beijing Fengtai Hospital from August 2017 to August 2022 were divided into 4 groups,with 50 cases in each group.In preoperative group,0.5μg/(kg?h)dexmedetomidine was intravenously pumped 30min before anesthesia induction,and the drug was stopped before the start of surgery.In intraoperative group,0.5μg/(kg?h)dexmedetomidine was infused intravenously after the beginning of the operation,and the drug was stopped 30min before the end of the operation.In postoperative group,0.5μg/(kg?h)dexmedetomidine was infused intravenously after the main steps of the operation,and the drug was discontinued at the end of the operation.In control group,the same volume of normal saline was injected intravenously 30min before anesthesia induction.The recovery quality,hemodynamics,inflammatory response,and adverse reactions were compared among the four groups.Results The extubation time of postoperative group[(43.84±5.12)min]was significantly longer than that of preoperative group[(16.73±3.28)min],intraoperative group[(18.05±3.47)min],and control group[(25.63±4.64)min],the difference was statistically significant(P<0.05).Ramsay sedation scores,mean arterial pressure(MAP),heart rate(HR),C-reactive protein(CRP),tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 were compared between preoperative group and intraoperative group before anesthesia induction in a quiet state(T0),immediately after extubation(T1)and 5min after extubation(T2).The difference was not statistically significant(P>0.05).Ramsay sedation scores and levels of CRP,TNF-α and IL-6 at T1 and T2 in postoperative group were significantly higher than those in preoperative group,intraoperative group and control group,and MAP and HR were significantly lower than those in preoperative group,intraoperative group and control group,with statistically significant differences(P<0.05).There was no significant difference in the overall incidence of adverse reactions among the four groups(P>0.05).Conclusion Intravenous infusion of dexmedetomidine at different time points in children undergoing craniotomy has no obvious adverse reactions,but intravenous infusion of dexmedetomidine before and during operation has little effect on hemodynamics and inflammatory response during anesthesia recovery period,and the quality of recovery and sedation is better.Intravenous infusion of dexmedetomidine after operation will prolong extubation time.

Objective To explore the differentially expressed micro RNAs (miRNAs) in serum-derived extracellular vesicles (EVs) of patients with Parkinson's disease (PD) and healthy individuals.Methods Thirty-six patients with late-onset Parkinson's disease (LOPD,age of onset being more than 50 years),20 patients with early-onset Parkinson's disease (EOPD,age of onset being no more than 50 years),and 10 healthy individuals were collected in our hospital from January 2015 to December 2017.Serum of all subjects was collected and EVs were extracted.The miRNA chip and real-time quantitative (qRT)-PCR were used to analyze and verify the differentially expressed exosomal miRNAs between PD patients and healthy individuals;Cytoscape software was used to analyze the common target genes regulated by differentially expressed miRNAs,and String database was used to analyze the interaction and function of target genes.And qRT-PCR was further used to analyze the expression level and correlation of differentially expressed miRNAs and their target genes in patients with EOPD and LOPD,respectively.Results As compared with healthy individuals,LOPD patients had significantly up-regulated expression levels ofmiR-143-3p and miR-1180 in the serum derived EVs,and statistically down-regulated miR-451 and miR-500 levels (P＜0.05).The miR-143-3p expression level in EOPD patients was significantly higher than that in LOPD patients (t=3.888,P=0.018).The target genes B-cell lymphoma-2 (BCL2) and AKT1 were involved in regulating the cholinergic synapses and neurotrophin signaling pathways,and the BCL2 relative expression level in EOPD patients was significantly higher than that in LOPD patients and healthy controls (P＜0.05);and the relative expression levels of BCL2 and miR-143-3p were negatively correlated in the serum derived EVs of EOPD patients (r=-0.556,P=0.025).Conclusion The miR-143-3p and BCL2 expression levels are negatively correlated in the serum derived EVs of EOPD patients;changes of expression level may be related to plasticity of synaptic structure and apoptosis of nerve cells.

Objective To investigate the clinical efficacy and safety of transcatheter arterial chemoemblization (TACE) using raltitrexed and lobaplatin in treating advanced hepatocellular carcinoma (HCC).Methods From March 2009 to November 2014,95 cases were treated by raltitrexed combined with lobaplatin (raltitrexed group) through TACE and 124 cases by fluorouracil combined with oxaliplatin (fluorouracil group) through TACE.Disease control rate (DCR),median progression-free survival (mPFS) time and median overall survival (mOS) time were compared between the two groups.Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis in SPSS 16.0.Results The disease control rate of raltitrexed group was 91.6％ (87/95),compared with fluorouracil group of 84.6％ (105/124) in fluorouracil group (x2 =2.505,P =0.474).The mPFS of raltitrexed group was 6.8 months and that of fluorouracil group was 5.9 months (x2 =5.542,P =0.019);mOS of raltitrexed group was 13.6 months and fluorouracil group was 11.4 months (x2 =5.953,P =0.015).The main adverse reactions in the two groups were not statistically significant (P ＞ 0.05).Conclusions TACE using rahitrexed and oxaliplatin prolongs the progression free survival and overall survival time of patients with advanced hepatic carcinoma.