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AIM: To explore whether tumor necrosis factor-α (TNF-α) induces necroptosis in murine long bone osteocyte-like cell line MLO-Y4 and the possible mechanism.METHODS: The MLO-Y4 cells were divided into control group, TNF-α group, TNF-α+necrostatin-1 (Nec-1) group, TNF-α+Z-VAD group and TNF-α+receptor-interacting protein 3 (RIP3)-siRNA group.The death rate of MLO-Y4 cells was assessed by flow cytometry with Annexin V-FITC/PI staining.The morphological features of the cells were observed under transmission electron microscope (TEM).The protein levels of RIP1, RIP3 and cleaved caspase-3 were determined by Western blot.Finally, the numbers of total cells and RIP1-RIP3-positive cells were observed under laser scanning confocal microscope.The production of reactive oxygen species (ROS) in the cells was measured by DCFH-DA staining.RESULTS: Compared with control group, the apoptotic or necroptotic rate of the cells induced by TNF-α was increased significantly (P<0.01).The increased apoptotic or necroptotic rate was dramatically reduced by treating with Nec-1, Z-VAD or RIP3-siRNA transfection (P<0.01).In TNF-α group and TNF-α+Z-VAD group, a lot of MLO-Y4 cells with typical necroptotic morphological features were observed under TEM.However, obvious necroptotic cells were not found in Nec-1 or RIP3-siRNA treatment group.The protein level of RIP1 in the cells treated with Nec-1 was sharply lower than that in TNF-α group (P<0.01).However, Z-VAD did not reduce the elevated levels of RIP1 and RIP3.RIP3-siRNA effectively down-regulated the protein level of RIP3 compared with TNF-α group (P<0.01).Nec-1 effectively down-regulated the protein levels of RIP1 colocalized with RIP3 compared with TNF-α group (P<0.01).However, Z-VAD did not reduce the levels of RIP1 colocalized with RIP3.Nec-1, Z-VAD and RIP3 siRNA significantly decreased the ROS levels (P<0.01).CONCLUSION: TNF-α induces the necroptosis of MLO-Y4 cells.RIP3 play vital roles in the cell necroptotic signal pathway.ROS may be the executor of necroptosis of MLO-Y4 cells.
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Objective To study the effect of laparoscopic neoplasty and changes of procalcitonin (PCT) and cell-mediated immunity in elderly patients with duodenal ulcer perforation. Methods Forty-four elderly patients with duodenal ulcer perforation were divided into laparoscopic neoplasty group (n=20) and open neoplasty group (n=24).The level of PCT was assayed by colloid immunization. Blood levels of CD3+, CD4+, CD8+and natural killer (NK) cells were detected by flow cytometry before operation and at 0.5 h, 24 h, 48 h and the 7th day after operation.Surgical complications and postoperative hospital stay were observed in two groups. Results There was no significant difference in the activity of PCT before operation between two groups. There were no significant differences in CD3+, CD4+, CD8+ and NK cells 0.5 h before operation between two groups. The level of PCT was significantly lower at 24 h and 48 h after operation in laparoscopic group than that in open operation group (P<0.05). There were significant differences in the serum levels of CD3+, CD4+, CD8+and NK cells with time changes in two groups (P<0.05). There were significant decreases in CD3+, CD4+and CD8+at 24 h after operation than those before operation in two groups, which were significantly lower in open operation group than those of laparoscopic group except for CD8+. The levels of CD3+, CD4+and CD8+recovered to the levels before operation at 48 h after operation in laparoscopic group. In open operation group CD3+did not return to the level before operation at 7 d after operation. The levels of NK cells were decreased at 24 h after operation in two groups, but there was no significant difference in the level of NK cells between two groups. The level of NK cells returned to preoperative levels at 48 h after operation in laparoscopic group, while there was no significant difference in the level of NK cells before and after operation in open neoplasty group. Compared with the open neoplasty group, there were fewer complications and shorter hospital stay in laparoscopic group. Conclusion The laparoscopic repair operation for gastric and duodenal ulcer perforation has a fewer effects on the stress response and immune function in elderly patients, and has the advantages in the protection of immune function.
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Objective To observe the clinical effects of hyperbaric oxygen combined large dose ganglioside on delayed encephalopathy after acute carbon monoxide poisoning(DEACMP).Methods A totol of 72 patients of DEACMP were randomly divided into the observation group(37)and the control group(35).The patients of control group were given the routine treatment with hyperbaric oxygen.In the treatment group patients were given large doses of gangliosides 100 mg,1 times a day,for a period of 20 to 30 days additionally to the routine treatment of hyperbaric oxygen.All patients were examined before and after treatment in EEG examination (EEG),mini-mental state examination(MMSE).Results The abnormal rate of EEG was 37.83％(14/37) in the control group,which was significantly lower than that of 65.71％(23/35) in the treatment group((x2 =5.60,P ＜ 0.05).In the treatment group,the MMSE was(15.45±2.93) and(23.70±2.13) before and after treatment,respectively; in the control group,the MMSE was(14.88±2.84) and(20.33±2.09) before and after treatment.The MMSE was significantly impoved after treatment in both group(t =3.18 and 2.91,Ps ＜0.05).Furthermore,the MMSE impoved more significantly in the treatment group compared to the control group (t =6.28,P ＜ 0.05).The effective rate of treatment group was 86.48％(32/37),which was significantly higher than that of 51.43％(20/35) in the control group(x2 =7.72,P ＜ 0.01).Conclusion Application of large dose ganglioside treatment had significant clinical effects in delayed encephalopathy after acute car-bon monoxide poisoning,which is better than the effect of routine hyperbaric oxygen treatment.
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Objective To investigate the efficacy and mechanism of hyperbaric oxygen combining conventional therapy on persistent vegetative state ( PVS).Methods Sixty-two cases of PVS patients were treated with hyperbaric oxygen ( HBO ) combinting with conventional therapy.( 1 ) Hyperbaric oxygen therapy:hyperbaric oxygen chamber was adopted and the air pressure was 0,18 -0.20 MPa.The patients breathed pure oxygen for 30 min,2 times per day with a 10 min interval,and continued for 10 days as a course with an average of 4 to 6 courses.(2)Conventional therapy:All patients received drug treatment in hyperbaric oxygen therapy at the same time,including hemostasis,dehydration,anti-infection,nerve cell nutrition agents,wake promoting agents and supportive therapy.All patients were divided to three groups according to the course of disease:26 - 59 days in A group,60-119 days in B group,120 -268 days in C group,and the relationship between disease course and HBO treatment efficacy was analyzed.Results Afer HBO combining conventional therapy,27 cases (43.5％) were recovered,18 cases(29.0％ ) had obvious effect,8 cases( 12.9％ ) had effect,9 cases( 14.5％ )were inefficacy.The efficiency related to the course of disease.The total efficiency rate was 85.5％.The efficacy of HBO treatment had significant difference in A,B and C group ( 96.2％,62.5％ and 41.7％ in A,B and C group respectively,x2 =24.83,P ＜ 0.01 ).The shorter course indicated the better efficacy.Conclusion Hyperbaric oxygen combining conven.
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Objective To explore the epidemic trend of influenza and pandemic virus variation in Maanshan, and to provide scientific basis for the prevention of influenza. Methods The ILI data of monitor hospitals and some ILI nasopharyngeal specimens were collected, then influenza virus was isolated by MDCK cells, and influenza serotypes were identified by HI. Results 15 904 cases of ILI were reported from 2006 to 2007. The detectable rate of influenza like illness in outpatients was 6.63%. 2 656 nasopharyngeal specimens of ILI were collected, and 358 strains of influenza virus were isolated, with isolation rate of 13.48%, of which 142 were H1N1 subset influenza A virus, 49 were H3N2, 72 were Victoria subset influenza B virus, and 96 were Yamagata. Conclusions During 2006 and 2007 two peaks of influenza-like illness in outpatients was in winter-spring and summer seasons of each year. More attention meeds to be paid to the monitoring of influenza.
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Objective To investigate the expression and significance of Ezrin and ICAM-1 in gastric carci- noma.Methods The expression of Ezrin and ICAM-1 were examined by S-P immunohistochemistry assays in 78 ca- ses of gastric carcinoma.30 cases of nonmalignant tissue and 30 cases of normal stomach.The relationship between them and their pathological characteristics were analyzed.Results ①The expression rates of Ezrin and ICAM-1 were obviously higher in carcinoma group than that of nonmalignant group and normal group(P<0.01),in the lym- phatic metastasis group than that in non lymphatic metastasis group(P<0.05),and in T3.T4 group and than that in T1,T2 group(P<0.05).②The expression level of ICAM-1 was positively related with that of Ezrin(r=0.46. P=0.00).③The test of the expression of Ezrin and ICAM-1 simutaniously could obviously increase the sensitivity and specificy for detecting the metastasis of stomach malignant tumor(P<0.05).Condusion The over expression of Ezrin and ICAM-1 is related to the pathogenesis and invasion as well as metastasis of stomach carcinoma.The combined detection of two markers can obviously increase the sensitivity and specificy for predicting metastasis of stomach carcinoma.
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Objective To explore and describe the sampling methods for aerosol from cooling towers in the centralized air conditioning system, early-warning indicators, the effect factor and present status of contamination by Legionella.Methods Air-conditioning systems were sampled two rounds in 13 guesthouses, markest and bathing spots of Maanshan City in June-August, 2008.Aerosols were collected by fluid-striking microbial aerosol sample collector, while water samples from cooling tower and swelling box were obtained.Legionella were determined respectively for 67 samples by germiculture and polymerase chain reaction.Results The positive rate of Legionella in aerosols by germiculture was 0, meanwhile it was 50.0% by polymerase chain reaction for Legionella pneumophila.The positive rate in aerosols was 90.0% as the air-sampling volume exceeded 400 L.The positive rate of aerosols increased while the operating-time of cooling tower increased.The relative humidity in Legionella positive groups was higher than that in Legionella negative groups.Conclusion The positive result for Legionella pneumophila by polymerase chain reaction should be an important indicator for early-warning for the occurrence and spreading of Legionnaires' disease.The air-sampling volume of over 400 L is available by sampling collector for the detection of Legionella pneumophila by polymerase chain reaction.Risk of Legionella contamination in aerosol would be raised in high-humidity environment.
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Objective To obtain genes encoding the novel molecules for diagnosis of schistosomiasis.Methods Juvenile S.japonicum cDNA library was immunoscreened to obtain positive clones.By DNA sequencing and sequence analysis,the target gene was amplified by PCR and subcloned into prokaryotic plasmid pET28a.The recombinant plasmid was transformed into E.coli BL21 followed by expression of the protein induced by IPTG.The protein was identified by Western blotting.Results 34 positive clones were obtained,24 of which were chosen to be sequenced,13 of which were Sj22600 gene.The protein were recognized with sera of infected rabbits and patients with acute and chronic schistosomiasis by Western blotting.Conclusion The gene coding for Sj22600 membrane protein was screened with high frequency in the cDNA library.The E.coli BL21 transformed with the recombinant plasmid can express the fusion protein,which shows immunoactivity.