Objective To investigate the effect of Celecoxib on human brain microvascular endothelial cells release6-keto-PGF1α,TXB2 and apotosis after irradiation.Methods The logarithmic growth phase cells were divided into control groups (Con),simple irradiation (IR) groups and combination groups (IR+C).CCK-8 and clone formation experiment were used to evaluate the effects of radiosensitivity and toxicity of celecoxib.The results were observed atthe time point of 6 h,12 h,24 h,48 h after irradiation.ELISA was used to test the contents of 6-keto-PGF1α and TXB2,which metabolized by PGI2 and TXA2 from culture medium after irradiation at different time points in different groups.TXB2/6-keto-PGF1αratios were calculated.Annexin V-FITC/PI double staining method was used to measure the apoptosis rates at different time points in different groups.Western blot was used to measure the protein expression.Paired t test difference.Results Compared with simple irradiation group,there were no significant radiosensitivity (SER=0.96) in combination groups incubated with30 μmol/L of celecoxib.Compared with the control group,the ratio of TXB2/6-keto-PGF1αincreased at each time point in IR and IR+C (P<0.05),and the apoptosis rates increased (P<0.05).Cox-2,P-JNK and Cleaved caspase-3 increased.Compared with IR,the ratio of TXB2/6-keto-PGF1αdecreased at each time point in IR+C (P<0.05),and the apoptosis rates decreased (t=3.34~6.38,P< 0.05).The protein expression of Cox-2,P-JNK and Cleaved caspase-3 decreased.Conclusions Celecoxib may help to protect HBMECs from releasing TXA2 and decreasing the ratio of TXB2/6-keto-PGF1α,and inhibitting apoptosis after irradiation.The mechanisms of apoptosis inhibition may be related to the inhibition of Cox-2 and P-JNK,caspase-3 Cleaved proteinexpressions.

Objective To describe the experiences when different methods were used to treat early-onset antibody-mediated rejection (AMR) after kidney transplantation.Methods The clinical data of 42 recipients who experienced early-onset acute AMR after kidney transplantation in our department from Jan.2010 to Apr.2016 were retrospectively analyzed.The recipients were divided into 3 groups based on different strategies against AMR:group A (plasma exchange with intravenous immunoglobin);group B (bortezomib solo),and group C (combination of bortezomib and sirolimus).Results All the AMR episodes were diagnosed by kidney biopsy 9-27 days after transplantation.The AMR reversal rate in groups B and C was significantly higher than that in group A (100％ versus 60.00％,P=0.034;100％ versus 60.00％,P=0.007).The AMR recurrence rate in groups B and C was significantly lower than that in group A (0 versus 41.67％,P =0.035;0 versus 41.67％,P =0.007).The recipient survival rate was 100％ in all the three groups.There were 11 graft losses in group A,and none in group B or C.The graft survival rate in group B at 6 months,1 year and 3 years was significantly higher than in group A (100％ versus 60.00％,P =0.034;100％ versus 55.00％,P =0.021;100％ versus 50.00％,P =0.013).The graft survival rate in group C at 6 months and 1 year was significantly higher than in group A (100％ versus 60.00％,P =0.007;100％ versus 55.00％,P =0.003).There was no significant difference in AMR reversal rate,AMR recurrence rate and graft survival rate between groups B and C.There was no significant difference in incidences of infection,hyperlipidemia and bone marrow suppression among the three groups.The incidence of diarrhea in groups B and C was significantly higher than in group A (50.00％ versus 0,P =0.001;42.86％ versus 0,P =0.001).The incidence of peripheral neuritis in group B was significantly higher than in group A (25.00％ versus 0,P =0.02),but similar to group C.There was no significant difference in average serum creatinine level among three groups within 1 year after treatment (P＞ 0.05).Antibodies against human leukocyte antigen (HLA) and donor specific antibodies were detected in all the 42 recipients before treatment.The negative conversion ratio of panel reactive antibody (PRA) in group A was significantly lower than in groups B and C (10.00％ versus 87.50％,P＜ 0.001;10.00％ versus 92.86％,P ＜ 0.001).The PRA recurrence rate in group A was significantly higher than in groups B and C (85.00％ versus 37.50％,P＜0.001;85.00％ versus 0,P＜0.001),while that in group B was significantly higher than in group C (37.50％ versus 0,P =0.014).The ratio of Treg in peripheral blood at 3-12 month after treatment in group C was significantly higher than in groups A and B (P＜0.05).Conclusion Treatment for early-onset AMR after kidney transplantation based on bortezomib might be an effective and safe strategy.Graft longterm survival might benefit from the combination of bortezomib and sirolimus.

Objective To discuss the efficiency and safety of conversion from tacrolimus(Tac)to cyclosporine A(CsA) in patients with new onset diabetes after transplantation (NODAT).Methods The glucose metabolism parameters and related clinical indicators in 45 Tac treated renal transplantation recipients who developed NODAT were retrospectively analyzed.The oral immunosuppressive strategy was Tac + mycophenolate mofetil (MMF) + prednisone(Pred).Results 32 cases were converted to CsA whereas 13 patients stuck to Tac.After conversion,fasting plasma glucose (FPG)decreased from(8.2 ± 2.7)mmol/L to(5.9 ± 1.2)mmol/L(P ＜ 0.01)and HbA1c level decreased from (7.0 ± 0.9) ％ to (6.1 ± 0.7) ％ (P ＜ 0.05).The level of FPG and HbA1c was lower in the conversion group than in the control group(P ＜ 0.05).During the 1-year follow-up,the curative rate of NODAT was 53.1％ (17/32) in the conversion group while it was 0％ in the control group.No acute rejection happened after the conversion.There was no obvious change in renal function.The 1-year survival rate of patient and the transplanted kidney was 100％.Blood pressure and lipid levels were stable after the conversion.Conclusion Conversion from Tac to CsA is a simple and effective strategy to improve glucose metabolism in renal transplantation recipients with NODAT.

Objective To describe the clinical characteristics,outcome and prognostic factors of primary intraspinal tumor treated with postoperative radiotherapy.Methods Thirty-three patients diagnosed as suffering from intraspinal tumor treated with postoperative radiotherapy were analyzed.There were eighteen male and fifteen female,with a median age of 37 years(range 7-64 years).Two patients were treated with grossly total resection,25 subtotal resection,2 biopsy only.These patients were treated either with(~(60)Co)(n =18,before 1998) or 6?MV-X ray(n =15,after 1998) to the total dose of 45-68?Gy.The overall response and survival rates were calculated by Kaplan-Meier method and the difference between groups was tested by the log-rank method.The prognostic factors related to the characteristics of patient,tumor,and treatment were determined.Results The overall response,five-year and disease-free survival rate was 81.3%,59.4% and 56.3%,respectively.By multivariate analysis,the histological type(P= 0.039) and gender(P= 0.049) had independent impact on the 5-year survival rate.Ependymoma had a better prognosis compared to other histological subsets and female gained more survival advantage over male.There was no close relationship between survival and other factors,such as age,extent of surgery,initial site,irradiation dose,or the interval between surgery and radiotherapy.Conclusions Postoperative radiotherapy is safe and effective for primary intraspinal tumor.The histological type and gender are important prognostic factors of survival in primary intraspinal tumors.

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BACKGROUND:The clinical study on the mechanical properties of biomaterials for cruciate ligament reconstruction is of great importance, which can provide a theoretical basis for the clinical development of materials with good structure, biomechanics and compatibility. OBJECTIVE:To analyze the mechanical properties of vascularized patelar tendon graft. METHODS:The anterior cruciate ligaments of the left foreleg from 42 New Zealand white rabbits were cut off. Then, these rabbits were randomly and evenly divided into two groups: experimental group and control group. Rabbits in the experimental group were implanted vascularized patelar tendon graft for anterior cruciate ligament repair. Rabbits in the control group were implanted with autologous non-vascularized patelar tendon graft for anterior cruciate ligament repair. At the 8th, 12th and 16th weeks after implantation, biomechanical properties of specimens from these two groups were detected. RESULTS AND CONCLUSION: There was no significant difference on the fracture location at different time points between these two groups; at the 12th and 16th weeks after transplantation, the knee laxity in the experimental group was higher than that in the control group (P < 0.05). At the 8th week after transplantation, there was no significant difference in the knee laxity between these two groups. At the 8th, 12thand 16th weeks after transplantation, the maximum load in the experimental group was higher than that in the control group (P < 0.05). At the 8th, 12th and 16th weeks after transplantation, the stiffness in the experimental group was higher than that in the control group (P < 0.05). There was no significant difference in the maximum tensile elongation between these two groups at different time points after transplantation. These results demonstrate that vascularized patelar tendon graft has good mechanical properties.

Objective To investigate the dynamic changes of VEGF mRNA and protein in the injured brain of rats irradiated by 20 Gy of electron beams.Methods A total of 114 Sprague-Dawley rats were randomly divided into two groups,42 rats in sham group with sham-irradiation,and the other 72 rats in irradiation group that the whole brains of rats were administered with a single dose of 20 Gy electron beam (6 MeV) to establish an animal model of radiation-induced brain injury.All rats were terminated at 1,3,7,14,28,42,56 d post-irradiation respectively.The pathological changes were observed with light microscopies.Real-time quantitative reverse transcriptase-PCR was used to measure the level of VEGF mRNA in the brain tissue.The expression of VEGF protein in whole brain tissue was detected with Western blot.The VEGF protein expression in brain endothelium,gliacyte and neuron were measured via immunohistoehemistry with computerized image processing.Results The pathological process of radiationinduced brain injury (RBI) included brain endothelial cell damage,brain edema,thrombosis formation and ablation,revascularization and angiogenesis.In the irradiation group,the expression of VEGF mRNA was declined significantly at 1,3,7,2g,42 d post-irradiation (t =16.275-46.118,P ＜ 0.05).The VEGF protein expression in whole brain tissue was up-regulated at 1 and 7 d after irradiation,but downregulated at 3,14,28,42,56 d post-irradiation.There were VEGF positive cells in the brain endothelium,gliacyte and neuron at 1 d after irradiation.The expression of VEGF protein in gliacyte and neuron significantly was increased at 1,14,42,56 d post-irradiation (t =-8.394--4.697,P ＜ 0.05),and increased significantly in brain endothelial cells at 1,14,and 42 d after irradiation (t =-5.554--4.159,P ＜ 0.05).Conclusions The expression of VEGF mRNA was relatively suppressed in RBI rats.The expression of VEGF protein in whole brain tissue was up-regulated only at the early stage of RBI.The expressions of VEGF protein in brain endothelium,gliacyte and neuron were increased in the observation period,which induced brain edema and the formation of thrombosis at the acute RBI stage,and participated in the progression of angiogenesis and thrombosis ablation at the early-delayed RBI stage.

Objective To evaluate the effect of penehyclidine hydrochloride pretreatment on pulmonary function during mechanical ventilation in the patients with chronic obstructive pulmonary disease (COPD) undergoing non-thoracotomy.Methods Sixty patients with COPD,aged 58-82 yr,weighing 45-76 kg,of ASA physical status Ⅱ or Ⅲ,scheduled for elective non-thoracotomy under general anesthesia,were randomly divided into Ⅰ,Ⅱ and Ⅲ groups (n =20 each) using a random number table.Anesthesia was induced with iv midazolam,sufentanil,cisatracurium and propofol.The patients were endotracheally intubated and mechanically ventilated.At 30 min before endotracheal intubation,normal saline 5 ml (group Ⅰ),penehyclidine hydrochloride 0.01 mg/kg (group Ⅱ) or penehyclidine hydrochloride 0.02 mg/kg (group Ⅲ) was injected intravenously.At 30,60 and 120 min of ventilation,airway peak pressure (Ppeak),airway plateau pressure (Pplat),airway resistance (Raw) and dynamic lung compliance (Cdyn) were recorded.At 120 min of ventilation,arterial blood samples were obtained for blood gas analysis and oxygenation index (OI),respiratory index (RI),physiologic dead space fraction (VD/VT) and alveolar-arterial oxygen gradient (A-aDO2) were calculated.At 30 min before ventilation and 120 min of ventilation,blood samples were drawn from the radial artery for determination of the serum concentrations of TNF-α,IL-8 and IL-10 by ELISA.The extubation time and pulmonary complications within 72 h after operation were recorded.Results Compared with group Ⅰ,Ppeak,Pplat,Raw,RI,VD/VT and A-aDO2 were significantly decreased,Cdyn and OI were increased,the serum TNF-α,IL-8 and IL-10 concentrations and incidence of pulmonary complications were decreased,and no significant change was found in the extubation time in Ⅱ and Ⅲ groups.There were no significant differences in the parameters mentioned above between group Ⅱ and group Ⅲ.Conclusion Penehyclidine hydrochloride pretreatment can reduce the inflammatory responses during mechanical ventilation,improve pulmonary function,and decrease postoperative pulmonary complications and is helpful for prognosis in the patients with COPD undergoing non-thoracotomy.

Objective To evaluate the effect of Fat Mass and Obesity Associated (FTO) gene on radiosensitivity of human glioma cell A172 and investigate its potential mechanism by changing the expression of FTO gene.Methods Cells were divided into five groups according to their FTO protein expression level.The normal expression group was recorded as A172 Group,the low-expression and its negative control group was A172/siRNA and A172/NC Group,and the over-expression and its negative control group was A172/FTO and A172/PC group.FTO protein expressions were assayed by Western blot in A172 Group after irradiation.Clonogenic assay was executed to evaluate the relationship between FTO gene and radiosensitivity.Immunofluorescence and Western blot assay were applied to detect the proteins of DNA damage and repair.Results FTO protein expression level in A172 Group was significantly related to the irradiation dose and the time post-irradiation.The radiosensitization ratio (SERD0) of A172/siRNA and A172/FTO group were 1.829 and 0.812 respectively.Not only the number of γ-H2AX foci increased (t =-21.884,P ＜ 0.05) in A172/siRNA 1 h post-irradiation but the decreases of p-p95/NBS1 and Ku80 proteins were also detected (t =24.731,23.293,P ＜ 0.05) together with the increase of Rad50 protein (t =3.140,P ＜ 0.05).But the expressions of these proteins in A172/FTO group were opposite to the above phenomenon (t =0.642,-8.364,26.829,P ＜ 0.05).Conclusions FTO gene is a radiation-resistant gene,which may because the regulation of FTO gene could alter the primary injury and DNA damage repair in the irradiated tumor cells.

Objective To investigate the effects of mechanicl ventilation with lower tidal volume and positive end-expiratory pressure (PEEP) on pulmonary function during laparoscopic surgery in patients with chronic obstructive pulmonary disease (COPD).Methods Forty patients with COPD,aged 60-82 yr,with body mass index of 16-29 kg/m2,undergoing elective laparoscopic surgery,were randomly divided into 2 groups (n =20 each) using a random number table:conventional ventilation group (group Ⅰ) and mechanical ventilation with lower tidal volume and PEEP group (group Ⅱ).Anesthesia was induced with midazolam,sufentanil,cisatracurium and propofol and maintained with iv infusion of propofol,cisawacurium and remifentanil.The patients were endotracheally ventilated and mechanically ventilated.In group Ⅰ,fresh gas flow was set at 2 L/min,VT at 10 ml/kg,and I∶E at 1∶2 during ventilation.In group Ⅱ,fresh gas flow was set at 2 L/min,VT at 6 ml/kg,I∶E at 1∶2 and PEEP at 6 cm H2O during ventilation.PErCO2 was maintained at 35-45 mm Hg in both groups.Airway peak pressure (Pp~),airway plateau pressure (Pplat),airway resistance (Raw) and dynamic lung compliance (Cdyn) were measured at 5 min after intubation (T1),45 min of pneumoperitoneum (T2),and 15 min after the end of pneumoperitoneum (T3).Arterial blood samples were obtained at T1,T2 and T3 for blood gas analysis.Alveolar-arterial oxygen gradiant (A-aDO2),oxygenation index (PaO2/FiO2),respiratory index (RI) and physiologic dead space fraction (VD/VT) were calculated.The extubation time and development of complications were recorded within 48 h after operation.Results Compared with group Ⅰ,Ppeak and Plat at T2 and Raw at T1,2 were significantly decreased,Cdyn at T2 and PaO2/FiO2 at T1-3 were significantly increased,RI,VD/VT and A-aDO2 were significantly decreased at T1-3,and the incidence of hyoxemia,atelectasis and rales was decreased within 48 h after operation in group Ⅱ (P ＜ 0.05).There was no significant difference in the extubation time between the two groups (P ＞ 0.05).Conclusion Mechanical ventilation with lower tidal volume (6 nl/kg) and PEEP (6 cm H2O) can improve the pulmonary function during laparoscopic surgery in patients with COPD.