BackgroundCorneal graft reject is a major cause of corneal transplantation failure.Although many immune-suppressing drugs have been utilized to reduce the reject response,their adverse effects on organ and tissue are still insoluble.The tolerance induction of natural killer T (NKT) cells is currently under investigation.However,the study on the application of NKT cells in high risk corneal transplantation is seldom.ObjectiveThe present study was to explore the effects of α-GalCer-activated NKT cella on allografts survival after high-risk corneal transplantation surgery via retro-bubar injection.Methods The lymphocytes were picked up from the spleen of SPF Lewis rats and cultured in RPMI 1640 medium with 100 mg/L α-GalCer.After one week,NKT cells were sorted by the FACSVantage system as CD161+ TCR-α+ cell from the lymphocytes with the cell densities 5×106/ml.Ten SPF Fisher344 rats were used to prepare the donor corneas,and 20 Lewis rats served as recipients.The high risk corneal transplantation models were created by corneal suturing in 20 recipient rats.Penetrating keratoplasty (PKP) was performed in the model rats.0.1 ml NKT cells or the same volume of normal saline solution were retro-bubarly injected at the end of surgery respectively.The corneal allografts were observed and scored based on Holland criteria at the three-day interval under the slit lamp for 30 days.Two weeks after surgery,three rats from each group were sacrificed by excessive anesthesia method and the eyeballs were obtained for histopathological examination.The inflammatory cell infiltration ( CD4+ and CD8+ ) in grafts was evaluated by immunochemistry and flow cytometry.The use of the animals complied with the Statement of ARVO.ResultsThe mean survival time of the allografts was (7.90± 1.37) days in normal saline solution group and (14.70± 1.49) days in NKT cell group,showing a statistically significant difference between the two groups ( t =10.61,P =0.00 ).Two weeks after surgery,all the allografts showed the severe opacity with lots of new blood vessels and edema in normal saline solution group.However,the corneal grafts were clear in NKT cell group.Abundant CD4+ and CD8+T lymphocytes were seen in the allografts in normal saline solution group,but the inflammatory cells were obviously less in NKT cell group.The percentage of NKT cells in the spleen was (5.67±0.25)％ in NKT cell group and ( 1.21±0.19)％ in normal saline solution group ( t =8.43,P =0.00 ).Conclusionsα-GalCer-activated NKT cells can prolong the survival time of allografts in high-risk corneal transplantation.Retro-bubar injection of α-GalCer-activated NKT cells probably is a new approach to the prevention of the rejection of corneal transplantation.

Objective To investigate the correlation between the expression of tumor O (6)-methylquanine DNA methyl-tranferase(MGMT) and pathological grade,and the influence of racial factors on tumor MGMT expression levels for glioma patients. Methods Compare and analysis the correlation between the pathological grade and MGMT levels and the racial factors on MGMT expression levels by the immunohistochemical staining on the tumor specimens of 33 Uygur and 61 Han. Results The positive rate of 61 Han gliomas pations with MGMT is 45.90％ and 33 cases of the Uygur is 30. 30％ , there's no clear correlation between the racial factors and the tumor MGMT levels. (P ＞ 0. 05). Comparative the 94 patients with pathological level and tumor MGMT level, there is no clear correlation between pathologic level and MGMT pression in tumor tissues (P ＞ 0. 05 ). ConclusionThere's no clear correlation of tumor MGMT expression and pathological levels; and there's no significant effect between racial factors and expression of glioma MGMT.

0.05),respectively.CONCLUSION:Intravenous drip combined with oral administration of Amoxicillin/Clavulanate Potassium can replace intravenous dip of Amoxicillin/Clavulanate Potassium bid.For those ?-Lactams drugs with short t1/2,the unreasonable intravenous drip of which qd can be replaced by the intravenous drip combined with N-time oral administration or with N-time intramuscular injection.

AIM: To study the protective effects of Naokangning (Rhizoma Chuanxiong, Radix salviae miltiorrhizae, Hirudo, etc.) against cerebral ischemia. METHODS: We observed the effect of Naokangning on mice's resistance to cerebral ischemia when bilateral common carotid arteries and vagus nerves were ligated and hypoxia under normal pressure and airtight circumstance; With the model of partial cerebral ischemia by blocking rats'middle cer ebral artery (MACO):the effects of Naokangning on the activity of superoxide dismutase (SOD) and creatine kinas e(CK), the content of malondialdehyde (MDA) and nitric oxide (NO) were measured. RESULTS: Naokangning significantly raised mice's ability of anti-cerebral ischemia and prolonged span of life in hypoxia, Moreover, it also obviously improved the activity of SOD, reduced content of MDA in cerebrum, content of NO and activity of CK in blood serum after ischemia. CONCLUSION: Naokangning could strikingly protect brain caused by cerebral ischemia.

Objective To compare the effects of different medications of fentanyl during anesthesia induction on fentanyl-induced cough.Methods Four hundred and twenty ASA Ⅰ or Ⅱ patients aged 18-60 yr undergoing selective operations under general anesthesia,were randomly divided into 4 groups ( n =105 each):group Ⅰ (control group) received successive intravenous injection of midazolam 0.05 mg/kg,fentanyl 2 μg/kg,propofol 2 mg/kg,and rocuronium 1 mg/kg,group Ⅱ (pre-injection group) received successive intravenous injection of midazolam0.05 mg/kg,fentanyl 0.5 μg/kg,propofol 2 mg/kg,rocuronium 1 mg/kg,and fentanyl 1.5 μg/kg,group Ⅲ (dilution group) received successive intravenous injection of midazolam 0.05 mg/kg,fentanyl 2 μg/kg (20 μg/ml),propofol 2 mg/kg,and rocuronium 1 mg/kg,and group Ⅳ (last injection group) received successive intravenous injection of midazolam 0.05 mg/kg,propofol 2 mg/kg,rocuronium 1 mg/kg,and fentanyl 2 μg/kg.Fentanyl concentration was 50 μg/ml in each group except group Ⅲ.Endotracheal intubation was performed 2 min after anesthesia induction.The incidence and severity of fentanyl-induced cough before intubation were recorded and the incidence of propofol-induced pain was recorded.Invasive arterial blood pressure (ABP) and heart rate (HR) were observed before induction (T1 ),immediately after induction (T2 ),at time of coughing (T3 ),and at time of endotracheal intubation (T4).Results ABP and HR had no significant differences at T1,T2,T3,and T4between the four groups (P ＞ 0.05).The incidence of propofol-induced pain had no significant differences between the four groups (P ＞ 0.05).The incidences of cough was 7.6％ in group Ⅱ,9.5％ in group Ⅲ,and 1.9％ in group Ⅳ,which were significantly lower than 35.2％ in group Ⅰ ( P ＜ 0.01).The incidence of cough in group Ⅳ was significantly lower than that in groups [ and Ⅲ (P ＜ 0.05).In the four groups,ABP and HR were significantly higher at T3 than that at T1 and T2 ( P ＜ 0.01 ).Conclusion Different medications of fentanyl including last injection,pre-injection,and dilution of fentanyl can significantly reduce the incidence of fentanyl-induced cough during anesthesia induction,and injection has the best effect.

Objective The study was designed to compare the ultrasound-guided technique for the cannulation of subclavian vein with the traditional technique using anatomic landmarks.Methods One hundred and twenty ASA Ⅱ or Ⅲ patients undergoing cannulation of subclavian vein before gastrp-intestinal tumor resection were randomly divided into 2 groups (n=60 each) according to the technique used for cannulation:ultrasound-guided group (group US) and anatomic landmark group (group AL).The puncture time,successful puncture and complications were recorded.Resulls The success rate was 100% in group US;while the cannulation failed in one patient in group AL.The rate of successful puncture at 1st attempt was 100% in group US but ouly 90% in group AL.The cannulation time was significantly shorter in group US than in group AL.The incidence of accidental puncture of subclavian artery,hematoma and pneumothorax was significantly higher in group AL than in group US.Conclusion The ultrasound-guided catheterization of the subclavian vein is superior to the landmark technique.

Objective To investigate the effect of lung protective ventilation regimen on regional cerebral oxygen saturation(rSO2)during one-lung ventilation(OLV)in elderly patients undergoing radical esophagus cancer resection.Methods Forty ASA Ⅰ-Ⅲ patients,aged 65-76 yr,weighing 45-75 kg,undergoing radical esophagus cancer reseclion,were randomly divided into 2 groups(n =20 each):conventional ventilation group(group CV)and prolective ventilation regimen group(group PV).Anesthesia was induced with midaaolam 0.05 mg/kg,sufentanil 0.4 μg/kg,rocuronium 1 mg/kg and propofol 1.5 mg/kg and maintained with 2％ sevoflurane and intermittenl iv boluses of rocuronium 0.5 mg/kg.Double lumen tube was inserted.Correct positioning was verified by fiberoptic broncboscopy.The patients were mechanically ventilated.In group CV,PEEP was set at 0,Vt was set at 10 ml/kg,and I:E was set at 1:2 during two-lung ventilation(TLV)and OLV.In group PV,PEEP was set at 5 cm H2O,Vt was set at 6 ml/kg,and I:E was set at 1:2 during TLV and OLV.PETCO2 was maintained at 35-40 mn Hg in both groups.Arterial blood samples were taken before induction of anesthesia,at 10 min of TLV and at 30 min of OLV for blood gas analysis.Qs/Qt was calculated and rSO2 was recorded at the same time.Low rSO2 (rSO2 score ＞ 3000％)was recorded during OLV.Results Compared with group CV,PaO2 and rSO2 were significantly increased,and Qs/Qt was significantly decreased at 30 min of OLV,and the incidence of low rSO2 was significanfly decreased in group PV(P ＜ 0.05).Conclusion Lung protective ventilation regimen can improve oxygenation,decrease intrapulmonary shunt,and reduce the occurrence of low rSO2 during OLV in elderly patients undergoing radical esophagus cancer resection.

OBJECTIVETo construct a recombinant lentiviral vector (pXZ208-BDDhFVIII) mediating B-domain-deleted human coagulation factor VIII (BDDhFVIII) gene and investigate its expression in HLF, Chang-Liver and MSC cells.

METHODSBDDhFVIII gene fragment was separated by endonuclease digestion and was cloned into the multiple cloning sites of pXZ208 to construct a recombinant lentiviral vector pXZ208-BDDhFVIII. Viral particles were prepared by means of three-plasmid cotransfection of 293T package cells by calcium phosphate precipitation. After infection, the coagulant activity of human FVIII in the culture medium of 293T, HLF, Chang-Liver and MSC cells was assayed by one-stage method. The gene transduction efficiency was assayed by flow cytometry (FCM). Furthermore, PCR was performed to test the integration of BDDhFVIII.

RESULTSThe infection rates of HLF, Chang-Liver and MSC were (74.52 +/- 7.57)%, (27.24 +/- 6.53)% and (42.34 +/- 5.84)% respectively. The activities of FVIII in supernatants of HLF, Chang-Liver and MSC were (54.1 +/- 5.6)%, (22.5 +/- 2.9)% and (12.5 +/- 2.7)% respectively. BDDhFVIII gene integration was detected in all the infected cells.

CONCLUSIONThe recombinant lentiviral vector pXZ208-BDDhFVIII was successfully constructed and efficiently integrated into target cells to express human FVIII activity in vitro.

Cell Line ; Factor VIII ; biosynthesis ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection

OBJECTIVETo study changes in the radiographic appearance during weight-bearing and non-weigh-bearing in hallux valgus, and to analyse the correlation between the elasticity of plantar soft tissue of hallux valgus and the pain under the metatarsal head.

METHODSFrom May 2012 to October 2012, 240 feet of 120 patients with hallux valgus were enrolled in the study. The degrees of the pian under the metatarsal head of all the patients were observed. AP and lateral X-ray films of feet were taken on the condition of weight-bearing and non-weight-bearing. So the hallux valgus angle (HVA), the inter-metatarsal angle between the first and second metatarsals (IM1-2), the inter-metatarsal angle between the first and fifth metatarsals (IM1-5), top angle of the medial longitudinal arch (TAOTMLA),and anterior angle of the medial longitudinal arch (AAOTMLA) were measured on the X-ray films. The differences of HVA, IM1-2, IM1-5, TAOTMLA and AAOTMLA between two groups were compared, and the correlation between the changes of IM1-2, IM 1-5, TAOTMLA, AAOTMLA and the degree of the pain under the metatarsal head were analysed.

RESULTSOne hundred and forty-eight feet had the pain under the metatarsal head. The IM1-2, IM1-5 and TAOTMLA increased on weight-bearing position compared with those on non-weight-bearing position, but the HVA and AAOTMLA decreased on weight-bearing position compared with those on non-weight-bearing position. There was a moderate relationship between the changes of IM 1-2,IM1-5 and the degree of the hallux valgus deformity, as well as the relationship between the different of IM1-5 and the degree of the pian under the metatarsal head.

CONCLUSIONThe degree of the collapse of the arch of foot with hallux valgus becomes serious with its deformity increasing. The pain under the metatarsal head of hallux valgus increases with the increased changes of IM 1-2,IM 1-5 and TAOTMLA. Analysis of the X-ray observation indexes of hallux valgus on weight-bearing position and non-weight-bearing position has important significance in evaluating the degree of the collapse of the arch of foot with hallux valgus,preventing and curing the the pain under the metatarsal head.

Adult ; Aged ; Female ; Hallux ; anatomy & histology ; diagnostic imaging ; physiopathology ; Hallux Valgus ; complications ; diagnostic imaging ; physiopathology ; Humans ; Metatarsalgia ; complications ; diagnostic imaging ; physiopathology ; Middle Aged ; Radiography ; Weight-Bearing ; Young Adult

Objective To investigate the correlation between the expression of tumor VEGF and pathological grade,and the influence of racial factors on tumor VEGF expression levels for glioma patients.Methods Compare and analysis the correlation between the pathological grade and VEGF levels and the racial factors on VEGF expression levels by the immunohistochemical staining on the tumor specimens of 33 Uygur and 61Han.Results The positive rate of 61Han gliomas pations with VEGF is 81.97％ and 33 cases of the Uygur is 60.61％,there's clear correlation between the racial factors and the tumor VEGF levels.(P ＜ 0.05).Comparative the two groups patients separately with pathological level and tumor VEGF level,there is no clear correlation in Uygu ( P ＞ 0.05 ) and opposite in Han( P ＜ 0.05 ).Conclusion There's significant effect of the racial factors on the positive of glioma's VEGF and the relation between the pathological grade and the expression levels.