Objective To explore effect of endogeneous gangliosides(Gls) and integrin ?2?1 on protein phosphotyrosine expression of pp125 focal adhesion kinase (pp125FAK) after adhesion of SK-N-SH neuroblastoma cells to collagen(Col).Methods SK-N-SH cell line with high expression of integrin ?2?1 was cultured in presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol(D-PDMP).Effect of endogeneous Gls,anti-?2 and anti-?1 monoclonal antibody on protein phosphotyrosine expression of pp125FAK during adhesion of SK-N-SH cells to Col were determined by immunoprecipitate and Western blotting.Results After 6 days,endogenous Gls in cells were almost depleted.Gls-depletion,anti?2 and anti-?1 monoclonal antibody were able to decrease pp125FAK expression of SK-N-SH cells adherent to Col respectively.GD2,the major component of neuroblastoma cell Gls could reco-ver pp125FAK expression to a certain degree.Conclusions Endogenous tumor Gls regulate protein phosphotyrosine expression of pp125FAK during adhesion of neuroblastoma cells to Col.It is suggested that tumor Gls may increase signal transduction of tumor cell integrin ?2?1 by increasing tyrosine phosphorylation of pp125FAK.

OBJECTIVETo explore the role of protein kinase D3 (PKD3) in the regulation of matrix metalloproteinases 7 (MMP-7) expression in prostate cancer cells.

METHODSPC-3 cells were either stimulated with 100 nmol/L PMA to activate PKD3 kinase activity, or transiently transfected with PKD3 siRNA, and the relative expression level of MMP-7 mRNA were analyzed by real-time PCR using 2(-delta delta Ct) method. MMP-7 mRNA levels were also analyzed and quantified in HEK293 cells with over-expression of wild-type PKD3, PKD3 knockdown (using PKD3 siRNA), or over-expression of wild-type PKD3 followed by PKD3 knockdown.

RESULTSMMP-7 mRNA expression in PC3 cells was significantly decreased after PMA-induced PKD3 kinase activation. In contrast, PKD3 knockdown by siRNA transfection markedly increased MMP-7 mRNA level (P<0.01). MMP-7 mRNA level in HEK293 cells was significantly decreased by PKD3 over-expression, whereas obviously increased by PKD3 knockdown. Down-regulation of MMP-7 mRNA level in HEK293 induced by PKD3 over-expression was rescued by PKD3 knockdown.

CONCLUSIONPKD3 may contribute to the malignant progression of prostate cancer cells through negative regulation of MMP-7 expression.

Cell Line, Tumor ; Down-Regulation ; Gene Knockdown Techniques ; Humans ; Male ; Matrix Metalloproteinase 7 ; metabolism ; Prostatic Neoplasms ; enzymology ; metabolism ; Protein Kinase C ; metabolism ; Signal Transduction

OBJECTIVETo investigate the influence of enteral administration of carbachol on the intestinal dysfunction of both severely burn patients and rabbits with partial intestinal ischemia/reperfusion (I/R) injury.

METHODSSeventy-five white rabbits were inflicted with I/R injury and randomized into intestinal I/R (I, n=25), carbachol [C, n=25, with 3g/L carbachol (3 mg/kg) injection into duodenum 1 h after SMA occlusion] and sham operation (SO, n=25, with SMA isolation but no occlusion) groups, and 5 other as normal controls. The blood flow of intestinal mucosa was detected before and after SMA occlusion or admission of carbachol. Changes in diamine oxidase (DAO), D-lactate, xylopyranose absorption, blue dextran discharging time were measured at 2, 4, 6, 8, 24, 48, 72 h after SMA occlusion. In addition, eight severe burn patients with TBSA of 84 +/- 12% were enrolled in the study, and carbachol (15 microg/kg) was administered to patients when abdominal distension or bowel sound was lower than 2 times/min, then the number of abdominal distension and bowel sounds per minute were observed.

RESULTSThe blood flow in intestinal mucosa of rabbits without SMA occlusion was (102 +/- 5) PU, reduced to (48 +/- 6) PU after SMA occlusion, and increased to (77 +/- 3) PU after injection of carbachol. The plasma DAO activity and D-lactic acid content in I group began to increase 4 hours after SMA occlusion, and they reached the peak 24 hours after SMA occlusion (4.63 +/- 0.27 U/ml, 7.9 +/- 2.4 mg/L) , after that they decreased gradually, but still higher than the normal value (0.89 +/- 0.14 U/ml, 2.0 +/- 1.1 mg/L, P < 0.05). In carbachol group, data showed the same trends as that in intestine I/R group with lower values, while no obvious changes were in sham operation group (P > 0.05). The content of D-lactic decreased dramatically 2 hours after D-lactic administration in both I and C groups, increased 6 hours after SMA occlusion, then decreased gradually, but it in C group was always higher than normal values, and little fluctuation was in sham operation group. There was no blue dextran discharge 2 hours after SMA occlusion. The discharging distance increased 6 hours later, but it was obviously shorter than the normal value 24 hrs after operation (P < 0.05) , then it returned to normal 48 to 72 hrs after operation. In the C group, blue dextran discharge was found immediately after its injection, with obvious increase in the discharging distance to peak value (43 +/- 6 cm) 6 hours after injury, and returning to normal (28 +/- 3 cm) gradually. In severe burned patients, the bowel sounds was (1.6 +/- 1.1) per minutes before carbachol administration, then increased dramatically to (6.9 +/- 1.7) per minutes 10 mins after administration, reached to a higher level 30 minutes after administration (8.3 +/- 2.4 ) times/min, and it maintained to (6.1 +/- 1.3) times/min 1 hour after administration. Abdominal distension was ameliorated 2 hours after carbachol administration, six patients were able to defecate.

CONCLUSIONEnteral administration of Carbachol can increase the blood flow of intestine mucosa, help to improve the movement, absorption and barrier functions of intestine, and ameliorate intestinal dysfunction in patients with severe burns.

Adolescent ; Adult ; Animals ; Burns ; drug therapy ; physiopathology ; Carbachol ; therapeutic use ; Disease Models, Animal ; Female ; Humans ; Intestinal Mucosa ; blood supply ; metabolism ; Intestines ; physiopathology ; Male ; Rabbits ; Reperfusion Injury ; drug therapy ; physiopathology

·AIM:To investigate the clinical effect of high-frequency electric lacrimal duct plasty and annular lacrimal drainage tube implantation combined with acupuncture on upper lacrimal duct obstruction. ·METHODS:A total of 86 patients (102 eyes) with upper lacrimal duct obstruction who were treated in our department from July 2013 to June 2016 were 26 males(28 eyes) and 60 females(74 eyes). The treatment was high-frequency electric lacrimal duct plasty and annular lacrimal drainage tube implantation, combined with traditional Chinese medicine acupuncture plus oral medicine made by our department for 30d, followed up for 6mo after surgery to observe the effect. ·RESULTS:A total of 86 eyes(84.3%) were cured in 102 eyes,12 eyes (11.8%) were improved, 4 eyes (3.9%) were ineffective,and the total effective rate was 96.1%. Intraoperative complications: 1)orbital hemorrhage in 2 eyes, maybe caused by vessel injury at the front of ethmoidal sinus, the blood was stopped after press, the operation went successfully after 5d; 2) lacrimal duct rupture in 3 eyes, all because of uncareful operation or weak compliance;3)the duct was hard to implanted in 3 eyes and iatrogenic sinus occurred, we suggested them went to superior hospital. Postoperative complications:1) lacrimal duct rupture without epiphora in 3 eyes(2.9%);2)the duct draw out in 5 eyes (3.9%), the duct was draw and fixed to nasal vestibule, in other 4 eyes (4.9%) with fully draw out,in one of the 4,the duct was replaced and in the rest 3 of the 4, no treatment; 3) irritation symptoms at inner canthus in 11 eyes (10.8%) with symptomatic treatment; 4 ) granulation tissue hyperplasia in 2 eyes (2.0%), one of them near the lacrimal punctum was cut and disappeared after tobramycin dexamethasone eye drops,one in the lacrimal ductile, the duct was draw and after tobramycin dexamethasone eye drops or compound neomycin sulfate eye drops the hyperplasia did not enlarge, but the lacrimal passage irrigation was failed. ·CONCLUSION:It is effective of high-frequency electric lacrimal duct plasty and annular lacrimal drainage tube implantation combined with acupuncture on upper lacrimal duct obstruction.

OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

OBJECTIVE: To study the clinical features of neuroblastoma (NB) and the factors influencing survival rate. METHODS: A total of 44 children with NB who were admitted from April 2016 to February 2020 were enrolled as research subjects. A retrospective analysis was performed on their medical data and follow-up data. RESULTS: The common clinical symptoms of these 44 children were fever (10/44, 23%), mass (9/44, 20%), abdominal pain (8/44, 18%), cough (7/44, 16%), pale complexion (3/44, 7%), claudication (2/44, 5%), and abnormal activity (2/44, 5%). According to the INSS stage, 2 children (4%) had stage I NB, 5 children (11%) had stage II NB, 5 children (11%) had stage III NB, and 32 children (73%) had stage IV NB. The mean follow-up time was (15.3±1.5) months, with a recurrence rate of 20% and an overall survival rate of 82%. Among the 44 children, 29 (66%) achieved event-free survival and 7 (16%) had survival with tumor. The univariate analysis showed that a pathological type of NB and an increase in serum neuron-specific enolase (NSE) decreased the overall survival rate of children with NB (P<0.05). CONCLUSIONS The clinical symptoms of children with NB are not specific at the first visit. Fever, abdominal pain, and mass are common symptoms, and there is a high proportion of children in the advanced stage. The pathological type of NB and an increase in serum NSE may be associated with a reduction in the overall survival rate of children with NB.
Child ; Humans ; Infant ; Infant, Newborn ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Neuroblastoma ; Phosphopyruvate Hydratase ; Retrospective Studies ; Survival Rate

OBJECTIVETo investigate the role of PKD3 in prostate-specific antigen (PSA) expression regulation in androgen-dependent prostate cancer cells and explore the mechanism.

METHODSLNCaP cells containing low level of PKD3 were transfected with pEGFP-C2 or pEGFP-PKD3 plasmid followed by dihydrotestosterone (DHT) treatment, and PSA mRNA level was analyzed by RT-QPCR using 2(-delta delta Ct) method. Wild-type or kinase-dead PKD3 plasmids, human androgen receptor plasmid pSVAR0, pMMTV-luc of AR luciferase reporter and renilla luciferase reporter pRL-SV40 were cotransfected into HEK293 cells, and after treatment with DHT for 24 h, the cells were harvested and AR transcriptional activity were determined by dual-luciferase reporter assay. The subcellular localization of endogenous PKD3 and AR and their colocalization induced by DHT were observed by confocal microscopy.

RESULTSPSA mRNA level triggered by DHT was significantly increased by overexpression of pEGFP-PKD3 in LNCaP cells compared with that in pEGFP-C2 control cells (P<0.001). AR transcription in response to DHT treatment was also significantly up-regulated by wild type PKD3 expression (P<0.001), but partially down-regulated by kinase-dead PKD3 mutant (P<0.01). Endogenous PKD3 and AR in LNCaP cells not only translocated from the cytoplasm to the nucleus, but also colocalized with each other after DHT stimulation.

CONCLUSIONElevated AR transcriptional activity and enhanced expression of PSA induced by PKD3 in response to DHT treatment suggest that PKD3 contributes to the proliferation and malignant growth of androgen-dependent prostate cancer cells.

Cell Line, Tumor ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms ; metabolism ; Protein Kinase C ; metabolism ; Transcriptional Activation ; Up-Regulation

Adult ; Aged ; Animals ; Anthrax ; epidemiology ; prevention & control ; veterinary ; Cattle ; Disease Outbreaks ; prevention & control ; Disinfection ; Dogs ; Female ; Humans ; Male ; Middle Aged ; Prevalence

Objective To improve the load resistance ability of students in teenager aviation schools and carry out specialized training for the body situation and shape of students.Methods We selected students from 14 teenager aviation schools and measured their height,weight,and the circumferences of necks,chests,upper-arms,waists,hips,and thighs.We compared the data of these students from different areas and the parameters were analyzed statistically by correlation analysis.Results Totally 411 students from 6 southern schools and 403 students from northern schools were investigated.Their average body height,weight and body mass index (BMI) were (172.3 ± 4.39) cm,(62.7 ± 7.3) kg,and (21.1 ± 2.23)kg/m2 respectirely.Students from northern schools were taller and heavier than those from southern schools (P ＜ 0.05).There was no difference between students from northern and southern schools in BMI.Compared with the domestic average levels of height,weight and chest circumference of teenagers aged 14 to 17,students of teenager aviation schools were higher,but there was no difference in weight or chest circumference.The average circumferences of the chest,neck,left upper-arm,right upper-arm,waist,hip,left thigh and right thigh were (83.1 ± 5.06),(34.7 ±3.07),(25.2±2.33),(25.5 ±2.30),(73.4 ±6.73),(89.7 ±5.89),(50.1 ±5.22),and (50.2 ±5.27)cm,respectively.According to the data,students from northern schools had a bigger chest circumference and thigh circumference but a smaller neck circumference and upper-arms circumference than those from southern schools (P ＜ 0.05).The correlation coefficient between BMI and the circumferences of the chest,neck,left upper-arm,right upperarm,waist,hip,left thigh and right thigh was 0.2201,0.1066,0.2466,0.2499,0.2291,0.2149,0.2299,0.2264,respectively.All of them were statistically significant (P ＜ 0.0001).Conclusion Load resistance related parameters of students in teenager aviation schools are not different from the domestic levels,which points to the urgent need for load resistance ability training for these students.Some parameters vary between areas.BMI is positively related to those load resistance parameters.Based on regional variations and regularities found in this study,specialized load resistance ability training could be made more efficient.

OBJECTIVE: To investigate the protective effect of isoflurane delayed preconditioning on myocardial ischemia reperfusion injury and the potential mechanism in rabbits. METHODS: Thirty New Zealand male white rabbits were randomly assigned to 3 groups: Control group; I/R group; and 2.0% isoflurane group. Isoflurane group was exposed to 2.0% isoflurane-100% oxygen for 2 hours. Control group and I/R group were exposed to 100% oxygen for 2 hours and served as untreated controls. Twenty-four hours later I/R group and isoflurane group underwent 40 minutes of coronary occlusion followed by 2 hours of reperfusion. Blood samples were taken from the arterial line at 20 minutes before the occlusion(T1), 20 minutes after the occlusion(T2), 40 minutes after the occlusion(T3), 1 hours after the reperfusion(T4), and 2 hours after the reperfusion(T5) to determine the plasma level of TNF-alpha. At the end of the reperfusion, infarct size and area at risk were defined by Evans and TTC staining. The heart was harvested and levels of the p38MAPK activity were determined by Western blot, and ultrastructures were observed under the electron microscope. RESULTS: The p38MAPK activity of isoflurane group was significantly lower than that of I/R group (P<0.05). Isoflurane significantly (P<0.05) reduced the infarct size(19.7%+/-2.8% in isoflurane group) of the left ventricular area at risk as compared with the controls (37.8%+/-1.7% in I/R group).The injury of I/R group was worse than that of isoflurane group under the light microscope. Isoflurane group had a lower level of TNF-alpha than I/R group. CONCLUSION Isoflurane can inhibit p38MAPK activity during myocardial ischemia reperfusion and modulate the cytokine expression, which may be one of the molecular mechanisms of isoflurane delayed preconditioning on cardioprotection.
Animals ; Ischemic Preconditioning, Myocardial ; methods ; Isoflurane ; pharmacology ; Male ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Myocardium ; ultrastructure ; Rabbits ; Random Allocation ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism