Objective To evaluate the changes in the expression of DJ-1 protein during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Fifty male Sprague-Dawley rats,weighing 220-280 g,were used in this study.Type 1 diabetes mellitus was induced by intraperitoneal streptozotocin 65 mg/kg and confirmed by fasting blood glucose ＞ 16.7 mmol/L.Forty animals with type 1 diabetes mellitus were randomly divided into 3 groups:diabetes group (group D,n =10),diabetic sham operation group (group DS,n =15) and diabetic I/R group (group DIR,n =15).Another 10 non-diabetic rats in which citrate buffer 6 ml/kg was injected intraperitoneally were served as control group (group C).Myocardial I/R was produced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in group I/R.At 120 min of reperfusion,5 rats were sacrificed and myocardial specimens were c(on)tained for determination of infarct size in groups DS and DIR,and 10 rats were sacrificed and myocardial specimens were obtained for microscopic examination and for determination of cell apoptosis,malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of DJ-1 and phosphatase and tensin homologue (PTEN) protein.Apoptotic index (AI) was calculated.Linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein was analyzed.Results Compared with group DS,the myocardial infract size was significantly increased in group DIR (P ＜ 0.05).Compared with group C,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in groups D,DS and DIR (P ＜ 0.05).Compared with groups D and DS,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in group DIR (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups D and DS (P ＞ 0.05).There was linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein and the correlation coefficients (r) were-0.734,0.593,-0.818,and-0.812 in turn.Conclusion Down-regulation of DJ-1 protein expression is involved in myocardial I/R injury in diabetic rats via decreasing anti-oxidative stress responses and upregulating PTEN protein expression.

Objective To investigate the analgesic efficacy and spinal neurotoxicity of intrathecal (IT) different doses of dexmedetomidine in rats. Methods Sixty male SD rats weighing 180-220 g were randomly divided into 5 groups ( n = 12 each): groupnormal control (group C); group IT normal saline (group N); different doses of dexmedetomidine groups received IT dexmedetomidine 0.75, 1.50 and 3.00 μg/kg respectively (groups D1.3). Paw withdrawal threshold to mechanical stimulation (PWMT)with yon Frey filaments and tail flick latency (TFL) to a thermal nociceptive stimulus were measured before (To, baseline) and at 30 or60 rin after IT dexmedetomidine or normal saline administration (T1, T2 ) and the percentage of the maximum possible effect ( MPE ) was calculated. Lumbar segment of the spinal cord ( L4-6 ) was removed for microscopic examination and determination of c-Fos expression (by immuno-histochemistry) at 7, 24 and 48 h after IT dexmedetomidine or normal saline administration. Results PWMT, TFL and the percentage of MPE were significantly increased after IT dexmedetomidine as compared with the baseline values at T0 in groups D1-3 ( P ＜ 0.05). PWMT was significantly higher at T1 and TFL and the percentage of MPE were higher at T2 in groups D1-3 than in groups C and N,and in group D3 than in groups D1,2 ( P ＜ 0.05). At 7,24 h after IT dexmedetomidine c-Fos protein expression was significantly higher in group D3 than in groups C and N( P ＜ 0.05). There was no significant difference in c-Fos expression at 48 h after IT dexmedetomidine between group D3 and groups C and N ( P ＞ 0.05 ). At 24 h after IT dexmedetomidine c-Fos protein expression was significantly higher in group D3 than in other 4 groups( P ＜ 0.05). Slight spinal cord injury was observed at 24 h after IT dexmedetomidine in group D3. Conclusion IT dexmedetomidine has antinociceptive effect. High dose dexmedetomidine IT can produce transient reversible toxicity to the spinal cord.

Objective To evaluate the effects of ischemic postconditioning on brain injury induced by myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Diabetes mellitus was induced by intraperitoneal streptozotocin 60 mg/kg and confirmed by blood glucose level ＞ 16.7 mmol/L.Thirty male Sprague-Dawley rats,weighing 220-280 g,in which diabetes mellitus was successfully induced,were randomly allocated into 3 groups (n =10 each) using a random number table:group sham operation (group S),group I/R and group ischemic postconditioning (group P).Myocardial I/R was induced by occlusion of the anterior descending branch of the left coronary artery in I/R and P groups.Group P received 3 cycles of 10 s reperfusion followed by 10 s ischemia at the end of myocardial ischemia.The rats were sacrificed at 120 min of reperfusion and the brains were removed for microscopic examination and for determination of cell apoptosis (by TUNEL) and expression of interleukin-6 (IL-6),IL-8,IL-10,glycogen synthase kinase-3 beta (GSK-3β) and phosphorylated GSK-3β (pGSK-3β) (by immuno-histochemistry).Apoptotic index was calculated.Results Compared with group S,apoptotic index was significantly increased,IL-6 and IL-8 expression was up-regulated,and IL-10 and pGSK-3β expression was downregulated in I/R and P groups (P ＜ 0.01).Compared with group I/R,apoptotic index was significantly decreased,IL-6 and IL-8 expression was down-regulated,and IL-10 and pGSK-3β expression was up-regulated in group P (P＜0.01).There was no significant difference in GSK-3β expression among the 3 groups (P ＞ 0.05).The pathologic changes were significantly attenuated in group P as compared with group I/R.Conclusion Ischemic postconditioning can attenuate brain injury induced by myocardial I/R in diabetic rats,and inhition of activity of GSK-3β may be involved in the mechanism.

Objective To compare the roles of Toll-like receptor 4 (TLR4)/NF-κB signal pathway in acute lung injury (ALl) induced by blunt chest trauma and by blunt chest trauma-hemorrhagic shock and resuscitation (double hits) in rats.Methods Forty male Sprague-Dawley rats,aged 8 weeks,weighing 240-280 g,were randomly assigned into 3 equal groups (n =10 each) using a random number table:sham operation group (S group),blunt chest trauma group (T group),and blunt chest trauma and hemorrhagic shock and resuscitation group (group THSR).Lung contusion was induced in anesthetized rats by dropping a 300 g weight onto a precordial protective shield to direct the impact force away from the heart and toward the lungs.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.At 6 h after the model was established,the arterial blood samples were collected for blood gas analysis and detection of tumor necrosis factor-alpha (TNF-α) concentrations in serum.Oxygenation index (PaO2/FiO2) was calculated.The rats were then sacrificed and pulmonary specimens were obtained for determination of TLR4 expression and NF-κB ac tivity (by immunohistochemistry and Western blot) in lung tissues and for microscopic examination.Results Compared with group S,PaO2 and PaO2/FiO2 were significantly decreased,PaCO2 and TNF-α concentrations in serum were increased,TLR4 expression was up-regulated,and NF-κB activity was enhanced in T and THSR groups (P ＜ 0.05).Compared with group T,PaO2 and PaO2/FiO2 were significantly decreased,PaCO2 and TNF-α concentrations in serum were increased,TLR4 expression was up-regulated,and NF-κcB activity was enhanced in THSR group (P ＜ 0.05).The histopathological damage to lung tissues was aggravated in THSR group as compared with T group.Conclusion The role of TLR-4/NF-κB signal pathway in ALI induced by blunt chest traumahemorrhagic shock and resuscitation (double hits) is significantly stronger than that in ALI induced by blunt chest trauma alone in rats.

Objective To investigate the mechanism of myocardial ischemia and reperfusion-induced acute lung injury (ALl) and protective effect of ischemic postconditioning.Methods Forty SD rats were allocated to sham group,myocardial ischemia/reperfusion group (reperfusion group),ischemic postconditioning group (postconditioning group),and ischemic postconditioning + phosphatase and tensin homolog deleted on chromosome ten (PTEN) inhibiting group (inhibitor group) according to the random number table,with 10 rats per group.Myocardial ischemia/reperfusion was induced by left anterior descending coronary artery occlusion.Postconditioning was performed within 1 minute before reperfusion consisting of 3 10 s cycles of reperfusion followed by 10 s occlusion.Lung was immediately removed 120 minutes after reperfusion for HE stain,immunohistochemical detection of inflammatory factors and apoptosis factors,TUNEL assay of cell apoptosis,and Western blot of protein kinase B (Akt),phospho-Akt (p-Akt),glycogen synthase kinase-3β (GSK-3β),and phospho-GSK-3β (p-GSK-3β).Results Down-regulated B-cell lymphoma-2 (Bcl-2) and IL-10 and up-regulated Bcl-2 associated X protein (Bax),cysteinyl aspartate specific proteinase-3 (Caspase-3),IL-6 as well as IL-8 were observed in other 3 groups compared with sham group (P ＜0.01).Moreover,down-regulated Bax,Caspase-3,IL-6,IL-8 as well as TUNEL and up-regulated Bcl-2 as well as IL-10 were observed in reperfusion group compared to postconditioning group and tensor group (P ＜ 0.01).No statistical differences were found among the four groups in levels of Akt,p-Akt,and GSK-3β,but level of p-GSK-3β was significantly down-regulated in reperfusion group compared to other 3 groups(P ＜ 0.01).Conclusion Development of ALI may relate to down-regulation of p-GSK-3β evoked directly by the release of inflammation factors in early period of myocardial ischemia/reperfusion and ischemic postconditioning may attenuate the condition.

Objective To explore and evaluate the application value of damage control theory in bilateral craniotomy operation of severe craniocerebral trauma.Methods Seventy patients with Glasgow coma scale score ≤ 5 scores were divided into 2 groups by random digits table method with 35 cases each:standard trauma craniotomy group (normal group),non-standard craniotomy and small dural window exposure strategy group (improved group).The clinical data were compared.Results There was no significant difference in postoperative cerebral tissue bone window embedded meal,large area cerebral infarction incidence rate between two groups (P ＞ 0.05).There was significant difference in the duration of surgery,blood transfusion amount within 24 hours,traumatic epileptic seizure within 1 month,the acute intraoperative encephalocele between normal group and improved group [(2.80 ± 0.63) h vs.(4.21 ± 1.04) h,(3.90 ± 1.02) U vs.(5.55 ± 1.32)U,14.3％(5/35) vs.48.6％(17/35),5.7％(2/35) vs.25.7％(9/35)] (P ＜0.05).Followed up for 6 months,good prognosis,moderate disability,severe disability,vegetative state,death was 4,4,8,6,13 cases in normal group and 8,9,5,4,9 cases in improved group,and there was significant difference (x2 =5.040,P =0.025).Conclusion Severe craniocerebral trauma bilateral craniotomy damage is bigger,damage control theory to guide the improved operation method,can improve the rescue efficiency.

OBJECTIVETo observe the clinical effect of Sizhi-Xifang in the improvement of postoperative ankle function.

METHODSFrom Jan. 2006 to Mar. 2008,49 patients were divided randomly into treatment group and control group. The treatment group included 16 males and 8 females with an average age of (41.3 +/- 13.2) years, involving 13 cases of tye A, 9 of type B and 2 of type C based on AO classification. The control group included 15 males and 10 females with an average age of (38.2 +/- 10.9) years, involving 12 cases of type A, 10 of type B, 3 of type C. Minimal invasive percutaneous plate were used in each group. In treatment group 24 cases were treated with Sizhi-Xifang after the incisions were healed.

RESULTSThere were no incision infections, flaps necrosis, bone and plate exposure after treatment in two groups. All patients were followed-up for 4 to 16 months with an average of 8.6 months. According to Johner-Wruhs evaluation standard, there were 7 cases in excellent, 12 good, 5 in fair in treatment group and in control group there were 5 cases in excellent, 7 in good, 10 in fair and 3 in poor. The comparison of effect between two groups had significant difference (P < 0.05).

CONCLUSIONSizhi-Xifang is helpful to improve the ankle postoperative function.

Adolescent ; Adult ; Aged ; Ankle ; physiopathology ; Bone Plates ; Female ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Recovery of Function ; Skin ; Tibial Fractures ; physiopathology ; surgery ; therapy ; Treatment Outcome ; Young Adult

Objective To investigate vascular endothelial growth factor-c (VEGF-C) and vascular endothelial growth factor receptor-3(VEGFR-3) mRNA expression, microvessels density (MVD) and lymphatic microvessels density (LVD) in human hepatocellular carcinoma and normal liver tissue. Try to illuminate the relationship among VEGF-C,VEGFR-3,MVD,LVD and the clinical pathological features of hepatocellular carcinoma. Methods Liver tissue of 60 cases definitely diagnosed as hepatocellular carcinoma and 20 normal cases were collected. VEGF-C and VEGFR-3 mRNA expression were examined by RT-PCR, MVD and LVD were examined by immunohistochemistry staining. Relationship between these indexes and clinical pathological features of hepatocellular carcinoma was also analysed. Results VEGF-C and VEGFR-3 mRNA expression, MVD and LVD in hepatocellular carcinoma were higher than those in normal liver tissue (P<0.01); In hepatocellular carcinoma tissue, expression of VEGF-C mRNA positively related with VEGFR-3 mRNA, MVD and LVD(P<0.01). VEGF-C and VEGFR-3 expression positively related with portal vein tumor thrombus, intrahepatal metastasis and lymph node metastasis (P<0.01). MVD positively related with portal vein tumor thrombus and intrahepatal metastasis (P<0.01). LVD positively related with lymph node metastasis (P<0.01). Conclusion VEGF-C and VEGFR-3 expression increase in hepatocellular carcinoma tissue. They might play roles in tumor invasion and metastasis by inducing angiogenesis and lymphangiogenesis.

Objective To investigate the regulation of immune function in patients with acquired immune deficiency syndrome (AIDS) by the secretion of T cell 1 (Th1) gamma interferon gamma (IFN-γ) in patients with acquired immune deficiency syndrome. Methods From January 2011 to January 2014, 20 cases of laboratory-confirmed AIDS patients were collected for research. Patients with AIDS received highly active anti-retroviral therapy (HAART group), while 20 healthy volunteers were selected as control group. Before receiving highly active anti-retroviral treatment in six months and 12 months , the patients with AIDS were collected in blood study of peripheral blood mononuclear cells (PBMC) in separated culture group and IFN-γ co-culture group for testing culture differential expression of cytokines and immune cells in solution. Results Before treatment in 6 months and 12 months , the supernatants in the cultured alone group and IFN-γ co-culture group by IL-12, IL-21 levels were significantly lower than that in the healthy group (P < 0.05); After treatment in 6 months and 12 months , the supernatants in the IFN-γ co-culture group by IL-12 and IL-21 levels were significantly higher than that in the cultured alone group (PP < 0.05). Before treatment in 6 months and 12 months,the level of IP-10 and CD4 + CD25 + Foxp3Treg cells in the cultured alone group and IFN-γ co-culture group, were significantly higher than the healthy group (P < 0.05); After treatment in 6 months and 12 months, the supernatant in the IFN-γ co-culture group was significantly higher than the average in the cultured alone group in IP-10; CD4 + CD25 + Foxp3Treg cells in the IFN-γ co-culture group were significantly lower than that in the cultured alone group (P < 0.05). Conclusion Patients with AIDS received highly active anti-retroviral therapy can improve the immunity. IFN-γ may further stimulate the secretion of immune cytokine in patients with AIDS.

Objective To explore the relationship between cervical lesions in patients with high -risk human papilloma virus(HPV)infection and serum interleukin -1 beta(IL -1 ),interleukin 2(IL -2),interleukin 10(IL -10).Methods 180 cases of cervical lesions were treated in our hospital from August 2013 to August 2015. Among them,105 cases were infected with HPV,and 75 cases were not infected by HPV.The serum levels of IL -2, IL -10 and IL -1βwere observed in two groups,and compared the levels of serum IL -2,IL -10 and IL -1βin patients with different DNA hr -HPV load,the correlation between DNA hr -HPV load and IL -1β,IL -2 and IL -10 levels was analyzed.Results The levels of IL -1β,IL -10,IL -2 were (0.85 ±0.23)ng/L,(182.35 ± 10.02)ng/L,(38.97 ±5.23)ng/L in patients with HPV infection,the levels of IL -1β,IL -10,IL -2 in patients without HPV infection were (0.62 ±0.18)ng/L,(305.42 ±11.13)ng/L,(25.18 ±3.16)ng/L.The levels of IL -1β,IL -10 were significantly higher in HPV group than in without HPV infection group,the level of IL -2 in HPV group was significantly lower than that of uninfected HPV group,and the differences were statistically significant (t =7.222,20.328,-7.558,all P <0.01).There were significant differences in the load of DNA hr -HPV between different pathological changes of cervical lesions,and the DNA hr -HPV load increased significantly with the increase of the degree of disease(t =6.214,19.097,33.906,6.952,6.274,all P <0.05).IL -1βand IL -10 levels from high to low were DNA hr -HPV load volume >1 000,100 -1 000 and <100,IL -2 levels from high to low were DNA hr -HPV load <100,100 -1 000 and >1 000.DNA hr -HPV load was positively correlated with serum IL -1βand IL -10 levels in patients with cervical lesions and HPV infection(r =0.452,0.422,P =0.035,0.019),and nega-tively correlated with IL -2 level(r =-0.398,P =0.027).Conclusion hr -HPV infection is closely correlated with serum IL -1β,IL -2,IL -10 levels in cervical lesions patients,IL -1,IL -2 and IL -10 can be used as impor-tant index for clinical monitoring.