Objective:To study the function and regulation of CREB signaling in rat hypothalamic neurons by exogeneous corticotropin-releasing hormone(CRH). Methods:Perifused primary cultures of rat hypothalamic neurons. Intracelluar free Ca2+ concentr ation([Ca2+]i) was assayed at 32℃ by fluorescence analysis,the coverslip was mounted in a Delta Scan spectrofluorimeter(Photon Technology International);and phospho-CREB by western blot. Results:The basal lower intracellular Ca2+ level of perifused primary cultures of rat hypothalamic neurons cultured on coverslipy was markedly increased by CRH,and significantly attenuated by application of nifedipine(Nif) or CP-154526. CRH may cause the increase of phospho-CREB in the neuron of hypothalamus,while CP-154526,Nif can significantly inhibit the production of phospho-CREB. Conclusion: CRH may excite hypothalamic neurons directly suggest that phospho-and[Ca2+]i,suggesting a role of CRH type 1 receptor in the activation of CRH gene expression.

Objective:To study the effect of astrocyte activating PKA signal pathway on the proligeration and differentiation of nerve stem cells.Methods:Cerebral cortex of 13-day embryo mice was taken and the nerve stem cells were cultured scatteringly. Astrocyte was used to stimulate the nerve stem cells,and H89,PKA recepter antagonist, was used to pretreat the nerve stem cells of the cerebral cortex, then the contents of PKA,nestin,and NSF in the nerve stem cells were determined using western blot. MTT was applied to determine and analyse the effect of horizontal cell on the proliferation and orientational dif- ferentiation of cultured nerve stem cells. Results:In the normal control group the content of NSF in the nerve stem cells was low;after astrocyte stimulation the nestin content decreased rapidly,while the content of NSF increased gradually. H8(915 ?mol/L) could markedly supress the increase of NSF content in the nerve stem cell stimulated by horizontal cells. Conclusion:Astrocyte can promote the proliferation and differentiation of nerve stem cells by activating PKA signal pathway of nerve stem cells.

Objective:To study the different modus operandi of posterior fossa decompression on function recovery of spine cord in patients with Chiari malformation and syringomyelia.Methods:Of 87 patients with Chiari malformation and syringomyelia,51 patients were selected and big craniectomy was performed in all cases.The effects of big craniectomy in 51 patients and mini craniectomy in 36 patients were compared.Results:Among the 51 patients,22 patients(43.1%) improved significanly in their clinical symptoms,10 cases were stabilized,and 5 cases deteriorated.Among the 36 patients,28 patients(77.8%) improved significanly in their clinical symptoms,and 3 cases were stabilized.Conclusion:Surgical treatment may fully ameliorate the clinical syndromes.Mini craniectomy and expansile reconstruction of posterior fossa is a preferred surgery for the treatment of ChiariⅠmalformation.

Objective To observe the effect of furosemide on stria vascularis cultivated in vitro from guinea pig, to explore the probable mechanism of furosemide ototoxicity.Methods 20 healthy pigmented guinea pigs was randomly divided into two groups: Furosemide group (n=16), control group (n=4). The stria vascularis explant were cultured for 24 hours using the explant-culture technique. Experimental groups were added into imediately different final concentration of Furosemide (60ug/ml?300ug/ml?600ug/ml?1250ug/ml?2500ug/ml) differently, and continuously cultivated 30 minutes or 90 minutes , then observed structure feature of the cultivated stria vascularis .Results Furosemide group did not show pathological changes such as edema of stria vascularis,shrinkage of the intermediate cells and enlargement of intercellular spaces. No significant changes were found in the stria vascularis structure features between furosemide group and control group. Conclusion Furosemide did not induce remarkable edema on cultivated stria vascularis from guinea pig.These findings suggested that furosemide might induce ototoxicity by indirect mechanism.

BACKGROUND: Up to now the pathogenesy of nonalcoholic fatty liver(NAFL) is unclear. Uncoupling protein (UCP) of mitochondrial membrane can make oxidative phosphorylation uncoupling in mitochondrium, and then result in decreasing the efficiency of ATP synthesis by mitochondrium and participate the progression of fatty liver by changing energy metabolism.OBJECTIVE: To observe the expressive rule of UCP-2 in NAFL rat models at different time phases.DESIGN: Randomized control animal trial.SETTING: Department of Gastroenterology, Research Institute of Surgery,Daping Hospital, Third Military Medical University of Chinese PLA.MATERIALS: The experiment was performed at National Key Laboratory,Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from November 2004 to December 2005. A total of 64 clean-grade Wistar adult male rats were selected. After being fed with common feed for 1 week, the rats were randomly assigned into model group and normal control group with 32 rats in each group. In each group there were 4 time points: feeding for 2, 4, 8 and 12 weeks, 8 ones at each time point.METHODS: The rats in model group received high fat diet (88% basic feed, 10% pork fat and 2% cholesterol) for 12 weeks. Those in normal control group received common feed in different cages (6 rats in each cage).MAIN OUTCOME MEASURES: Blood was collected from anesthetic rats of the two groups at corresponding time point. Animals were killed,and then liver was extracted rapidly. ①Change of expression of UCP-2in liver was determined with immunohistochemical method and Western blot technique. ②Activity of triacylglycerol, free fatty acid (FFA) and alanine aminotransferase (ALT) in serum was measured with biochemical method. ③Content of malondialdehyde (MDA) in liver tissues was measured.RESULTS: Totally 64 rats were involved in the result analysis. ①Biochemical indicators in serum: Levels of triacylglycerol, FFA and ALT in serum were significantly higher in the model group than in the normal control group at weeks 4, 8 and 12, especially at weeks 8 and 12 (P ＜ 0.01). ②The expression of UCP-2 enhanced gradually with the exacerbation of fatt yliver in the model group, while it was negative in the normal control group.③Content of MDA in liver tissues: It was significantly higher in the model group than in the normal control group at weeks 4, 8 and 12, especially at weeks 8 and 12 [(4.07±0.15), (4.93±0.14), (5.20±0.20) μmol/g; (3.14±0.20), (3.12±0.18), (3.13±0.16) μmol/g,P ＜ 0.01].CONCLUSION: With the formation and aggravation of nonalcoholic fatty liver, levels of triacylglycerol, FFA and ALT in serum and content of MDA in liver tissues increase, and the expression of UCP-2 also increase. It is indicated that high fat diet can induce expression of hepatocyte UCP-2,decrease of ATP progression in cells and accelerate the formation and development of fatty liver.

Objective To evaluate the clinical outcomes of cemented bipolar femoral head arthroplasty in treatment of intertrochanteric fractures in elderly patients combined with heart,lung,brain and other diseases.Methods The study comprised 23 cases,aged 75-86 years old,treated with cemented bipolar artificial femoral head arthroplasty between January 2005 and October 2013 (replacement group).Evans' classification of the fracture was type Ⅰ in 3 cases,type Ⅱ in 9,type Ⅲ a in 5,type Ⅲ b in 2,type Ⅳ in 3 and reverse intertrochanteric fracture in 1.Additionally,23 cases with the similar physical condition treated with anatomical locking plates were chosen as controls.For comparisons between groups,operation time,amount of bleeding,drainage volume,mean time to extract drainage tub,antibiotics dosage,perioperative complications,complications of lying in bed,fracture complications and Harris score were measured.Results There were no significant differences between replacement and control groups with respect to operation time (52.7 minutes vs 52.0 minutes),amount of bleeding (160.9 ml vs 156.5 ml),drainage volume (162.2 ml vs 158.3 ml),Harris score (73.7 points vs 74.2 points),time to extract drainage tub (69.9 hours vs 68.9 hours) (P ＞ 0.05).Both groups had 5 days of preventive antibiotic usage.Better results were observed in replacement group than in control group concerning mean time of ambulation (8.1 days vs 109.3 days),rate of perioperative complications (19％ vs 59％),rate of bed rest complications (0 vs 36％) and rate of internal fixation complications (0 vs 18％) (P ＜ 0.05).Conclusion Cemented bipolar femoral head arthroplasty is effective to prevent the complications of internal fixation,bed rest as well as perioperation and improve the quality of life during the treatment of intertrochanteric fracture in elder patients with heart,lung,brain and other diseases.

Objective To explore the over-expression of the CCAAT enhancer binding proteinα(C/EBPα)in hepatic stellate cells (HSC-T6) which activated by protease-activated receptor 1(PAR1) ,to explore its possible relationship with cell proliferation ,cell activation and collagen secretion of HSC-T6 cell .Methods HSC-T6 cells were divided into 4 groups ,control group ,SFLLR group;vehicle plasmid+SFLLR group ,C/EBPαplasmid+SFLLR group .The eukaryotic vector harboring the full length of C/EBPαcD-NA was transfected into PAR1 activated HSC-T6 cell ,then Western blot was applied to detect the expression of α-smooth muscle actin(α-SMA) and collagen I and evaluate the effect of transfection on proliferation of HSC-T6 by MTT .Results The expression of α-SMA and collagen I in C/EBPαplasmid+SFLLR group were dramatically decreased compared with other 3 groups ,as well as proliferation after 48 h(P<0 .05) .Conclusion Over-expression of C/EBPαgene by transfection had inhibitory influence on prolif-eration ,activation and collagen secretion of HSC-T6 which were activated by PAR1 .

OBJECTIVETo explore the clinical effects of negative pressure wound therapy (NPWT) combined with artificial dermis grafting and autologous skin grafting on repair of open joint wounds and/or wounds with exposed bone fracture.

METHODSEleven patients with open joint wounds and/or wounds with exposed bone fracture, hospitalized from November 2008 to November 2014, were enrolled in the study. According to the differences of the first stage treatment, all patients were divided into experimental group ( n = 6, including 4 patients of open joint wounds, 1 patient of wound with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture), and control group ( n 5, including 2 patients of open joint wounds, 2 patients of wounds with exposed bone fracture, and 1 patient of open joint wound with exposed bone fracture). After debridement, the wounds in both groups were grafted with punctured artificial dermis, while NPWT was only used over the artificial dermis of experiment group for 1 week. In the operation at sacsod stage, autologous split-thickness skin was grafted on the vascularized artificial dermis in both groups. Results In 5 patients of open joint wounds in experimental group, the artificial dermis was vascularized well, autologous skin grafts survived, and wounds were healed. In 3 patients of open joint wounds in control group, the artificial dermis grafting all failed due to local infection, and then these wounds were repaired with local tissue flap grafting. Artificial dermis in 3 patients of wounds with exposed bone fracture in both groups was vascularized well after grafting, and the wounds were healed after autologous skin grafting, whether or not NPWT was used.

CONCLUSIONSNPWT combined with artificial dermis grafting and autolognus skin grafting can be used for repairing open joint wounds and/or wounds with exposed bone fracture.

Debridement ; Dermis ; transplantation ; Humans ; Negative-Pressure Wound Therapy ; methods ; Skin Transplantation ; methods ; Skin, Artificial ; Surgical Flaps ; Wound Healing

Objective To assess the significance of elevated serum N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in predicting 30-day mortality rate for patients in intensive care unit (ICU).Methods A retrospective study was conducted.The clinical data of 480 patients admitted to ICU in Tianjin First Center Hospital from June 2012 to October 2013 were studied.The characteristics of patients were recorded,the acute physiology and chronic health evaluation Ⅱ (APACHE I) score was calculated,and the routine blood tests,blood biochemistry,and serum NT-proBNP level within 24 hours after admission were determined.Patients were divided into non-survival and survival groups according to 30-day outcome,and the characteristics and clinical parameters were compared between two groups,and single factor analysis affecting prognosis was analyzed.All patients were divided into four groups according to four percentile method as NT-proBNP＜280 ng/L (group Q1),280 ng/L≤NT-proBNP＜ 1 200 ng/L (group Q2),1 200 ng/L≤NT-proBNP＜1 990 ng/L (group Q3),1 990 ng/L≤NT-proBNP＜4 700 ng/L (group Q4).A Cox regression analysis was used to test the independence of the predictors.A receiver operator characteristic curve (ROC curve) was plotted to compare the power of APACHE Ⅱ score and NT-proBNP in predicting the outcome.Kaplan-Meier analysis was used to assess survival rate among different NT-proBNP patients.Results By single factor analysis it was shown that age,APACHE Ⅱ score,creatinine,NT-proBNP and severe infection were those influencing the outcome.NT-proBNP 1 200-4 700 ng/L (groups Q3 and Q4) and APACHE Ⅱ score ＞20 were independent risk factors for predicting outcome [relative risk (RR) for group Q3 was 3.624,95％ credibility interval (95％CI) was 1.258-10.138; RR for group Q4 was 5.059,95％ CI was 3.202-9.993; RR for APACHE Ⅱ score ＞20 was 2.987,95％ CI was 1.345-5.823].The area under the ROC curve (AUC) for APACHE Ⅱ score and serum NT-proBNP was similar,and it was 0.831 (95％CI 0.778-0.884) and 0.876 (95％CI 0.827-0.925),respectively.The cut-off value of NT-proBNP for predicting mortality was 1 250 ng/L.Kaplan-Meier analysis showed that groups of Q2,Q3 and Q4 had lower cumulative survival rate compared with group Q1 (26.5％,25.4％,16.2％ vs.29.6％),and group Q4 showed most significant extent of lowering of survival rate (x2=122.920,P＜ 0.05).Conclusion Elevated NT-proBNP level in ICU patients signify severity of the disease,and it is an independent predictor of death of ICU patients.

Objective To investigate the expressions of brain-derived nerve growth factor (BDNF) and growth-associated protein 43 (GAP-43) in the hypothalamus of rats inflicted with restraint stress and their relationship with behavioral changes.Methods Forty male SD rats were divided into control group,restraint stress 7-day group,restraint stress 14-day group,restraint stress 21-day group according to the random number table,with 10 rats per group.Behavior changes were observed by open-field test,serum levels of corticotrophin releasing hormone (CRH) by enzyme-linked immunosorbent assay,and expressions of BDNF and GAP-43 in the hypothalamus by western blotting.Results Restraint stress 7-day group exhibited increases in spanning lattice times (50.0 ± 7.0),standing times (11.4 ± 2.1)and modification times (11.2 ± 2.7) compared with all other groups (P ＜ 0.05).Restraint stress 14-day group and restraint stress 21-day group showed significant decreases in spanning lattice times (35.5 ±7.5,29.4 ± 6.8),standing times (7.8 ± 4.9,5.6 ± 3.9) and modification times (6.7 ± 2.9,4.4 ±2.6) compared with control group (42.6 ± 5.4,8.9 ± 4.3,and 7.9 ± 3.0) (P ＜ 0.05).Restraint 14-day and 21-day groups showed significant increases in serum CRH level [(750.73 ± 123.68) pg/ml and (793.06 ± 115.84)pg/ml] compared with that in restraint stress 7-day group [(500.48 ± 88.71)pg/ml,P ＜0.05],but all were lower than (336.72 ±45.34) pg/ml in control group (P ＜0.05).Levels of BDNF and GAP-43 in the hypothalamus were the lowest in control group (0.672 ± 0.185 and 0.694 ±0.253).However,restraint stress increased the expressions of BDNF and GAP-43 in the hypothalamus,with the highest level in restraint stress 21-day group (1.357 ± 0.524 and 1.486 ± 0.679) (P ＜ 0.05).Conclusion Restraim stress can up-regulate BDNF and GAP-43 proteins in the hypothalamus,and lead to plasticity changes that may relate to stress-related behavior.