Objective To investigate the function of helper T lymphocyte cell(Th)1/Th2 cytokine in food allergy development.Methods A total 110 Balb/c mice were randomly divided into 2 groups:control group(40 mice)and food allergy group(70 mice).Food allergy animal models were established by ovalbumin,performed by skin prick test;in positive reaction mice,serum specific IgE,IL-4 and IFN-? were measured by enzyme linked immuosorbent assay(ELISA),and intestinal pathology were performed,and the mRNA expressions of IL-10,TGF-? in intestinal were measured by real-time PCR assay.Results In food allergy group,the mRNA expression of IL-10,TGF? in intestinal decreased(P

Objective To investigate the effects of vitamin E (VE) on renal ischemia/reperfusion (I/R) injury of rats.Methods A total of 18 male Wistar rats were randomly divided into sham-operated group,I/R group,VE + I/R group,and each group of 6 rats.All the animals were killed at the end of 24 h of reperfusion.Nephridial tissue were examined by light microscopy,and the level of blood urea nitrogen(BUN) and serum creatinine (SCr) were measured.The protein expressions of tumor necrosis factor α (TNF-α) were detected by Western blotting.Results Compared with sham-operated group,tubulointerstitial pathological injury in I/R group was significantly aggravated,which was shown by HE and PAS stain.Compared with I/R group,the degree of morphological changes as well as renal dysfunction in VE + I/R group were obviously lessened.Meanwhile,the levels of BUN,SCr in I/R group,VE + I/R group were (10.13 ± 2.14) mmol/L and (7.67 ± 1.63) mmol/L,(80.33 ±7.15) μmol/L and (63.67 ±5.40) μ mol/L,significantly higher than those in shamed-operated group ((3.85 ± 0.21) mmol/L,(48.67 ± 3.61) μmol/L;P ＜ 0.05).And the level of BUN and SCr in VE + I/R group were significant lower than those in I/R group(P ＜0.05).Western Blotting showed that the protein expressions of TNF-α in VE + I/R group were obviously lower compared with those in group of I/R without VE treatment (P ＜ 0.05).Conclusion Vitamin E can attenuate over-expressions of TNF-α in kidney following I/R,thus protect against structural damages and renal dysfunction in I/R rat models.

OBJECTIVETo summarize the clinical manifestation and diagnosis of ganglioneuroma in spine and investigate the clinical effect of surgical treatment.

METHODSThe clinical data of 6 patients underwent a surgery for ganglioneuroma in spine from January 2008 to January 2015 were retrospectively analyzed. There were 4 males and 2 females, aged from 2 to 63 years old with an average of 34.6 years. The courses of disease were from 3 days to 17 years. Five patients complicated with superficial hypesthesia in correlative level of tumor, and the muscle strength under tumor plane had decreased at different levels, with the strength of grade II-IV. Two cases complicated with hypermyotonia and positive bilateral Hoffmann's and Babinski sign. Five cases were sporadic lesion in correlative spinal canal and one case complicated with the giant occupying lesion in thoracic cavity.

RESULTSSix operations had been performed including 5 en bloc and 1 subtotal resection. Postoperative pathological results showed tumor cells scattered or fasciculated inserted into Schwann cells in the stroma. In 2 patients complicated with radiculalgia before operation, 1 case was relieved and 1 was invariant after operation. All 4 patients with preoperative dyscinesia in the limbs obtained improvement after operation. All the patients were followed up from 0.3 to 6.8 years with an average of 2.5 years. At the final follow-up, according to ASIA grade, 5 cases were good and 1 case was invariant. During the follow-up, only 1 patient experienced chemoradiation because of merging ganglioneuroblastoma and receiving subtotal resection. No recurrence in other 5 cases.

CONCLUSIONGanglioneuroma is a benign and rare tumors in spine. Clinically, radicular pain and sensory-motor disorders are the main manifestations. Its diagnosis depends on pathological examination. Prognosis of surgical treatment is good.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Ganglioneuroma ; diagnosis ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spinal Neoplasms ; diagnosis ; surgery

Objective To investigate the effect of intermedin (IMD) on the expressions of angiogenesis-related genes induced by renal ischemia reperfusion injury (IRI).Methods Wistar rats were randomly divided into four groups: control group,IRI group,empty plasmid group and IMD plasmid group.One week after removing the right kidney,eukaryotic expression vector encoding rat IMD gene was transfected into the left kidney using an ultrasound-microbubble mediated system.Renal IRI model was induced by clamping left renal arteries for 45 minutes followed by reperfusion for 1 d,2 d,3 d,4 d,7 d and 14 d.The expressions of hypoxia inducible factor-1α (HIF-1o),vascular endothelial growth factor (VEGF) and angiopoietin receptor Tie-2 were examined by RT-PCR and Western boltting.Results Compared with control group,an increase in HIF-1α,VEGF and Tie-2 was observed in the IRI group at d 1,d 2 and d 3 (allP＜0.05).The expression of HIF-1o peaked at d 1 d(P＜0.05),while VEGF and Tie-2 at d 2 (P＜0.05),followed by a decrease that was similar to the control levels at d 4(P＞0.05).Compared with the IRI group,the expressions of HIF-1α,VEGF and Tie-2 of IMD group were much higher and all reached the peak at d 1 (P＜0.05),maintained at d 2-4 (P＜0.05),followed by a decrease at d 7(P＞0.05).The above indexes had no differences between empty plasmid group and IRI group(P＞0.05).Conclusions IMD pretreatment may play an important role in the process of repair and regeneration after renal ischemia reperfusion injury byimproving the expressions of angiogenesis-related genes(HIF-1α,VEGF and Tie-2) induced by renal ischemia reperfusion injury.

Objective To investigate the effects of intermedin (IMD) pretreatment on associated repairing genes of the rats with injured kidneys by ischemia reperfusion injury (IRI)during the repair and regeneration process.Methods A total of 144 healthy male Wistar rats were randomly divided into 4 gourps:sham opration group,IRI group,empty plasmid group (EP)and IMD group.After resection of right kidneys of the rats,plasmid was transfected into the left kidneys by using ultrasonic microbubble technology.After one week,the renal IRI models were programmed.The samples of renal tissues after 1 d,2 d,3 d,4 d,7 d and 14 d of reperfusion were harvested respectively,then the mRNA expressions of the Pax-2,ZO-1,Ncam,Wt-1 and vimentin in renal tissues were detected by RT-PCR; the protein expressions of Pax-2,Wt-1 and vimentin were analyzed by Western blotting and the protein expressions of ZO-1 and Ncam were measured by ELISIA.Results (1) Compared with the sham group,the mRNA and protein expression of Pax-2,ZO-1,Ncam,Wt-1 and vimentin of the rats in IRI group increased significantly from day 1 to day 3 (P＜0.05),which peaked at day 2.After day 4,the above expressions in IRI group returned to normal level.(2) In IMD group,the mRNA and protein expressions of Pax-2,Zo-1,Ncam,Wt-1 and vimentin were significantly higher than those in other 3 groups (P＜0.05) at the same time after IRI day 1 to day 4,with a maximum in day 2.(3) The above expressions in IRI group,EP group,IMD group had no significant differences compared with sham group after day 7 or day 14 respectively (P＞0.05),and either between IRI group and EP group,though the expressions of the genes in IRI group and EP group increased compared with sham group after day 4.(4) The above expressions between IRI group and EP group also had no significant difference (P＞0.05).(5) Changes of ZO-1 and Ncam protein expression detected by ELISA were similar to those abore mRNA and protein expression by RT-PCR and Western blotting.Conclusion IMD pretreatment plays an important role in up-regulation of the expressions of associated repair genes during the process of repair and regeneration after renal ischemia reperfusion injury.

Objective To investigate the protective effect of intermedin(IMD)on renal ischemia reperfusion injury(IRI)and its mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups: control group, IRI group, empty plasmid group and IMD group. After remove of right kidney, plasmid was transfected into the kidney by ultrasonic microbubbles technology, and IRI model was made after 1 week. Renal pathology was observed by PAS staining. Renal tissue superoxide dismutase(SOD), myeloperoxidase(MPO), caspase-3 activity, and malondialdehyde(MDA)content were detected by colorimetric method. The intercellular cell adhesion molecule-1(ICAM-1), endothelin 1(ET-1)and P-selection expression of renal tissue were detected by immunohistochemical method. Apoptosis of renal tubular cell was detected by TUNEL.Results Compared with control group, tubulointerstitial pathological injury was significant aggravated in IRI group(P<0.01);compared with IRI group, IMD pretreatment significantly alleviated the degree of renal injury(P<0.01). Compared with control group, in IRI group, SOD activity was significantly decreased(P<0.05), MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 were increased significantly(all P< 0.01). Compared with IRI group, IMD pretreatment significantly increased SOD activity(P <0.05), decreased the MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 (all P<0.01). The apoptosis rate of renal tubular epithelial cells in IRI group was significantly higher than that in control group(34.83%±8.75% vs 3.33%±0.47%, P<0.01), while the apoptosis rate of IMD group(20.67%±7.71%)was significantly lower than that of IRI group. There was no difference of above indexes between empty plasmid group and IRI group. Conclusions IMD pretreatment protects against renal IRI. The mechanism may be at least partly related to the clearance of oxygen free radicals, the improvement of lipid peroxidation, inflammatory cell infiltration and cell apoptosis, leading to the decrease of the production of reactive oxygen species caused by oxidative stress.

Objective To investigate the effect and mechanism of adrenomedullin (AM) on apoptosis of renal tubular epithelial cell in rats induced by renal ischemia reperfusion injury. Methods Thirty-two Wistar rats were randomly divided into 4 groups: control group, IRI group, empty plasmid group and AM group. One week after removing the right kidney, eukaryotic expression vector encoding rat AM gene was transfected into the left kidney using an ultrasound-microbubble mediated system. After 1 week the transfer efficiency was detected by immunohistochemical method . Renal IRI model induced by clamping left renal arteries for 45 min followed by reperfusion for 24 h. Tubular cell apoptosis was detected by TUNEL assay. Bcl-2, Bax and Fas expressions were examined by RT-PCR. The expressions of caspase-3, caspase-8 and caspase-9 were determined by Western bolt analysis. Results The expression of AM in the AM group was significantly higher than the empty plasmid group (0.51±0.09 vs 0.23±0.05; P<0.05). Compared with the control group, the apoptosis rate of renal tubular cell in the IRI group was significantly higher [(38.79±7.52)% vs (2.89±0.52)%; P<0.05]. The expressions of Bax, Bcl-2, Fas, caspase-3, caspase-8 and caspase-9 were also significantly increased (0.72±0.18 vs 0.23±0.04, 0.80±0.12 vs 0.38±0.06, 1.24±0.25 vs 0.39±0.09, 0.76±0.13 vs 0.38±0.08, 0.92±0.14 vs 0.32±0.06, 0.89±0.12 vs 0.42±0.09; P<0.05). Bax/Bcl-2 was also significantly increased (0.91±0.18 vs 0.61±0.08; P<0.05). Compared with the IRI group, AM pretreatment significantly decreased the apoptosis rate of renal tubular cells [(19.36±6.78)% vs (38.79±7.52)%; P<0.05]. AM inhibited the up-regulation of Bax, Fas, caspase-3, caspase-8 and caspase-9, while promoting the up-regulation of Bcl-2 (0.48±0.11 vs 0.72±0.18, 0.62±0.07 vs 1.24±0.25, 0.53±0.08 vs 0.76±0.13, 0.46±0.08 vs 0.92±0.14, 0.51±0.12 vs 0.89±0.12, 1.23±0.25 vs 0.80±0.12; P<0.05). Bax/Bcl-2 significantly decreased (0.44±0.12 vs 0.91±0.18; P<0.05). The above parameters had no significant diffe-rence between the empty plasmid group and the IRI group (P>0.05). Conclusion AM can reduce apoptosis of renal tubular epithelial cell induced by renal IRI, the mechanism of which might be achieved by inhibiting caspase-dependent intrinsic and extrinsic pathways.

Objective To investigate the effect of intermedin (IMD) on renal ischemia/ reperfusion (I/R) injury after the up-regulation of IMD. Methods A total of 24 healthy Wistar male rats were randomly divided into four groups,sham-operated group,I/R group,IMD gene transfection +I/R group and empty plamid +I/R group.All the animals were killed at the end of 24 h of reperfusion.Histological changes and renal function were estimated.The expression and site of IMD were determined by Immunohistochemistry method,semi-quantitative RT-PCR and Western blotting.The protein expressions of endothelin 1 (ET-1),tumor necrosis factor αt (TNF-α) were detected by Western blotting. Results Compared with sham-operated group,tubulointerstitial pathological injury was significant aggravated in I/R group (7.6±2.3) and empty plamid +I/R group (7.0±1.8),and such injury was improved in IMD+I/R group (1.5±0.8) (P＜0.05).Compared with I/R group and empty plamid +I/R group,the renal dysfunction of IMD +I/R group was obviously lessened [BUN:(7.73±1.03) mmol/L vs (10.13±2.14) mmol/L,(9.77±1.92) mmol/L; Scr:(58.50±3.27) μmol/L vs (80.33±7.15) μmol/L, (75.67±7.58) μmol/L,all P＜0.05].IMD expression was weak in the plasma of tubulointerstitial cells in sham-operated group,and was up-regulated in I/R group. Compared with I/R group, immunohistochemical IMD expression increased obviously (262.03±67.89 vs 175.57±48.06,P＜0.01).The mRNA expression of IMD in IMD+I/R group was up-regulated significantly by 60.7％,66.1％ and the protein expression of IMD in IMD+I/R group increased significantly by 51.4％,55.9％ as compared to I/R and empty plasmid +I/R group.Meanwhile,the protein expressions of ET-1 and TNF-αt in IMD+I/R group were obviously lower compared with those in I/R group (ET-1:0.08±0.02 vs 0.17±0.02; TNF-α:0.21±0.04 vs 0.35± 0.02,all P＜0.05). Conclusion IMD gene transfected into kidneys of rats prior to I/R surgery can attenuate the over-expressions of both ET-1 and TNF-o in I/R injured rat kidneys as well as the damages to the structure and function of the kidneys.

Objective To explore the mechanism of protecting cells from hypoxia/ reoxygenation (H/R) injury by constructing specific small interference RNA (siRNA) to inhibit NALP3 expression in rat renal tubular epithelial cells (NRK-52E).Methods (1) To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 1 h,2 h,4 h,8 h,16 h and 24 h.The activity of lactae dehydrogenase (LDH) in the culture medium,cell count and cell viability,the expression of NALP3 were determined by biochemical method,trypan blue exclusion and Western blotting.(2) The siRNA was transfected into NRK-52E.The irrespective siRNA transfected group wasused as control.NALP3 expression was examined by Western blotting.(3) The cells were divided into 4 groups:control group,H/R group,irrespective siRNA transfected group and NALP3-siRNA transfected group.To establish the H/R injury model of NRK-52E by regulating the pressure of N2 in incubator to hypoxia condition,the cells were cultured with hypoxia for 1 h and then with reoxygenation for 4 h.And the expression of NALP3 was determined by Western blotting.(4)Cellular apoptosis was examined by Annexin V/PI staining and flow cytometry.NF-κB DNA binding activity,IκB-α,Bcl-2 and Bax expression were examined by EMSA and Western blotting.Results (1)Compared with the control group,the activity of LDH significantly increased,cell count and cell viability significantly decreased (all P＜0.05).The expression of NALP3 significantly increased and peaked at 4 h after H/R.(2)The specific siRNA could efficiently inhibit NALP3 expression in NRK-52E.Compared with the irrespective siRNA transfected group,the protein expression of NALP3 was significantly down-regulated in NALP3 siRNA transfected group (P＜0.05).(3)After hypoxia 1 h and reoxygenation 4 h,the activity of LDH and the expression of NALP3 increased.Compared with the irrespective siRNA transfected group,LDH concentration in media and the expression of NALP3 significantly decreased in NALP3-siRNA transfected group.(4)After hypoxia 1 h and reoxygenation 4 h,NF-κB DNA binding activity was increased,IκB-α phosphorylation and degradation,Bcl-2 and Bax were significantly up-regulated.However,compared with the irrespective siRNA transfected group,NF -κB DNA binding activity,IκB-α degradation and Bax/Bcl-2 were significantly decreased (P＜0.05) in NALP3-siRNA transfected group.At the same time,the ratio of apoptosis was significantly increased in three groups than that in control.Compared with the irrespective siRNA transfected group,the ratio of apoptosis in NALP3-siRNA transfected group was significantly decreased (P＜0.05).Conclusions H/R induces the expression of NALP3 in NRK-52E.The synthesized siRNA can inhibit the expression of NALP3 and protect NRK-52E from hypoxia/reoxygenation injury.The mechanism may be via inhibiting the activation of NF-κB,modulating expression of Bcl-2 and Bax,as well as decreasing cell apoptosis.

Objective To investigate the expressions of intermedin /adrenomeduliin 2 (IMD/AM2) and its receptor calcitonin receptor-like receptor (CRLR) in the kidney of rats after renal ischemia reperfusion injury(IRI). Methods Male Wistar rats were divided randomly into two groups: sham group and operation group. Renal IRI model was induced by clamping both renal arteries. Blood and kidney were harversted at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h after reperfusion, respectively. Renal histological changes were semi-quantitated. Expressions of IMD and CRLR in the kidney were detected by Western blot, and the content of IMD in serum was measured by radioimmunity at 12 h, 48 h, 72 h after repeffusion. Results Kidneys of renal IRI model rats displayed significant pathologic changes, and the changes were much severer at 48 h after reperfusion. The expressions of IMD and CRLR in kidney were significantly up-regnlated at 12 h, 48 h, 72 h after renal IRI (P<0.01). The level of IMD in serum increased at 12 h, 48 h, 72 h after renal IRI (P<0.05). Conclusion The expressions of IMD and its receptor are up-regulated in the kidney after renal IRI, which may participate in the pathophysiological changes induced by renal IRI.