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.

Acupuncture Therapy ; instrumentation ; Adult ; Female ; Humans ; Lactation ; Mastitis ; physiopathology ; therapy ; Young Adult

Objective To discuss human papilloma virus(HPV)prevalence and HPV genotypes distribution and the infection factors,to provide scientific guidance for the prevention and treatment of cervical cancer.Methods From March to November in 2009,605 women received cervical HPV testing in the Tumor Hospital of Harbin Medical University,to obtain specimens of cervical cytology,rapid flow-through hybridization technique (namely Hybribio flow-through hybridization)was used to detect HPV genotypes simultaneously.Single-factor and multivariate factors non-conditional Logistic regression analytic method was used to discuss the relationship between HPV infection of females and age,marital condition,level of education,level of income,occupation,initial age for sex,contraception,number of pregnancies,delivery approach and smoking.Results HPV infection rate was 21.49%(130/605),the positive rate of HPV infection in high-risk subtypes was 15.70%(95/605),the most common type was 5.29%(32/605)in the samples.Single factor non-conditional logistic regression model analysis showed that initial age for sex was the risk factor(X2=4.4618,P<0.05),HPV prevalence increased with a lower initial age for.sex reduced.But there was no significant difference in age,marital condition,education,income,occupation,contraception,number of pregnancy,delivery approach and smoking teams(X2=0.0525,1.8510,1.0348,0.2592,1.1176,1.5664,2.8835,1.4597,2.6161,all P>0.05).The analysis of multivariate factors nonconditional Logistic regression showed that the age of initially having sex,marital status and number of pregnancies were the risk factors(X2=21.6637,8.0574,15.7573,all P<0.05).Conclusions The risk factors for HPV infection are mainly about having sex too young,marital status and number of pregnancies,attention should be paid to screening for HPV.

OBJECTIVETo investigate the protective effects of Yuyin Ruangan Decoction(YRD, traditional Chinese medicine) on experimental hepatic injury in mice.

METHODSThe mice were randomly divided into control group, model group and YRD low, middle and high dose group(n = 11). By ip injection of D-GalN, CCk or thioacetamide (TAA), three models of hepatic injury mice were established to investigate the effects of YRD through detecting the indexes of liver function in serum and, the content of antioxidant system in the hepatic tissue.

RESULTSYRD could decrease the content of alanine aminotransferase (ALT)and aspartate aminotransferase (AST) in serum and that of malonaldehyde (MDA) in the hepatic tissue, upregulate the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the hepatic tissue. Furthermore, the above effects were dosedependent in a certain degree. CoNCLUSION: YRD has some protection effects on the model of experimental hepatic injury in mouse.

Alanine Transaminase ; blood ; Animals ; Antioxidants ; metabolism ; Aspartate Aminotransferases ; blood ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Liver ; drug effects ; enzymology ; Malondialdehyde ; metabolism ; Mice ; Superoxide Dismutase ; metabolism

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.

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 To determine the conditioning regimen suitable for mice allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Twelve BALB/c mice were randomly divided into 2 equal groups to undergo X-ray irradiation by linear accelerator at the dose of 7.0 Gy (pure X-ray group) or 60Co source irradiation at the dose of 7.0 Gy (pure γ-ray group).Thirty mice were randomly divided into 2 equal groups to undergo X-ray irradiation and then infusion of bone marrow from donor mice via caudal vein (X-ray + transplantation group) or γ-ray and then infusion of bone marrow via caudal vein (γ-ray + transplahtation group).3,5,7,10,15,20,and 30 d later peripheral blood samples were collected to calculate the number of white blood cells (WBCs) and detect the chimeric rates of lymphocytes by flow cytometry.5,10,and 20 d after irradiation 15 mice were killed with their lung,liver,small intestine,spleen,and femurs taken out to undergo pathological examination.Results The survival rates during the period 5-15 days of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group.The pathological changes of organs of the X-ray +transplantation group were all more severe than those of the γ-ray + transplantation group.Since the fifth day after transplantation cells originating from the donor began to appear in the peripheral blood.The chimeric rate of the γ-ray + transplantation group 10 days after transplantation was (95.53± 2.57) %.The chimeric rates 5,10,and 20 days after transplantation of the γ-ray + transplantation group were all significantly higher than those of the X-ray + transplantation group (t = 15.263,3.256,P < 0.05).The WBC count of both irradiation groups decreased to the lowest level 5 d later and began to increase 10 days after transplantation and the WBC counts of the γ-ray + transplantation group 10 and 20 days aftertransplantation were both significantly higher than those of the X-ray + transplantation group (t = 3.624,6.695 ,P < 0.05).The chimeric rats of the peripheral lymphocytes 10 and 20 days after transplantation of the γ-ray + transplantation group were both significantly higher than those of the X-ray + transplantation group (t = 12.317,8.295,P < 0.05).The homogeneity rate of transplantation of the γ-ray +transplantation group was better than that of the X-ray + transplantation group.Conclusions As a conditioning regimen in allogeneic hematopoietic stem cell transplantation γ-ray irradiation causes milder injury and accelerated reconstitution of hematopoiesis and immunity,in comparison with X-ray irradiation.

Objective: To investigate whether lipopolysaccharide induced parkin expression and mitophagy in macrophages.Methods:The murine peritoneal primary macrophages were aseptically isolated from Kunming mice and cultured in complete medium.The mitochondrial membrane potential of macrophages was detected by flow cytometry,after the cells were stimulated with 200 ng/ml LPS and labeled mitochondria with JC-1.The parkin mRNA level of macrophages was detected by RT-PCR, protein levels of parkin and autophagic related protein LC3 Ⅱ and LC3 Ⅰ were determined by Western blot.The distribution and co-localization of parkin with LC3 and mitochondria in macrophages were respectively observed by laser scanning confocal microscope, before and after the cells were treated with LPS.Results: Flow cytometry results after JC-1 staining showed that mitochondrial membrane potential in macrophages was declined after stimulation with 200 ng/ml LPS, and continuously decreased with prolonged treatment time.The mRNA levels of parkin were increased slightly within 6 h after LPS stimulation,but parkin proteins were increased significantly within 6 h after LPS stimulation.The results of parkin distribution showed that parkin was evenly distributed in the cytoplasm at normal status, but became the obvious punctate distribution after LPS stimulation in macrophages.Western blot results showed LC3 Ⅱ/LC3 Ⅰ levels were increased after LPS stimulation, indicating the appearance of macrophage autophagy.Confocal microscopy showed that there were co-localization of parkin,LC3 and mitochondrial in macrophages after LPS stimulation.Conclusion:Parkin expression is increased significantly and mediated mitochondrial autophagy in macrophages after LPS stimulation, which is involved in the clearance of damaged mitochondria,thereby playing a role in regulating macrophage inflammatory response.

We have designed and carried out problem based learning (PBL) pedagogy since 2004. According to clinical eases, students learnt the pathophysiology of heart failure knowledge by themselves. Each group recommen-ded one student to make an oral presentation and wrote a review about heart failure. Preparing clinical cases and group discussions are very important. At the same time we should pay attention to the change in role of the teacher in PBL and cooperation with other disciplines.

Objective:To study the effect of nerve growth factor(NGF)on bone fracture healing of inflicted T_(10)spinal cord injury(SCI)complicated with tibia fracture in rats.Methods:Totally 120 rats were randomly divided into tihia fracture group (F group,n=40),T_(10)SCI+tibia fracture group(FS group,n=40),and T_(10)SCI+tibia fracture+NGF group(FSN group,n=40).Four weeks later,the fracture sites in the 3 groups were subjected to CT scanning;the maximum transverse diameter of the fracture ends and the gray scales of non-osseous area were measured;the changes of biomechanics property of the fracture ends were determined by three-point bending test;the bone morphometry,bone density,and histomorphology of callus were determined;the expression of OCN was detected by immunohistochemical method;the osteoblast ultrastructure was observed by TEM and the expression ofⅠ,Ⅱtype collagen were examined by Western blotting.Results:The maximum transverse diameter of F group was less than those of FS group(P