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.

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 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.

Background Biometry of the anterior ocular segment parameter is very important for the diagnosis and treatment of glaucoma and ocular injury as well as measurement of intraocular lens(IOL).Objective This study was to compare the differences in the anterior chamber depth(ACD) and the central corneal thickness (CCT) between Sirius and Pentacam and evaluate the agreement of these two measurement methods.Methods The ACD and the CCT of 38 right eyes from 38 health volunteers aged 23- 32 years were measured with both Pentacam and Sirius.Three times of measurement were pedormed on each eye for each method to obtain the average values.The repeatability and agreement from each method were assessed as intraclass correlation coefficient( ICC ) and coefficient of variation(CV) and the agreement between these two methods were evaluated using Bland-Altman mode.ResultsThe mean ACD value was( 3.18±0.21 ) mm from Pentacam with the ICC 0.995 and CV 0.066.The mean ACD value from Sirius was (3.22 ±0.21 )mm with the ICC 0.996 and CV 0.065.The difference value in ACD between two methods was 0.04 mm,showing a significant difference( t =-6.225,P＜0.05 ) and a positive correlation (r=0.977) between two methods.The 95％ limit of agreement was( -0.04-0.13)mm within 1 standard difference (SD) of the mean value( ±0.21mm),which was acceptable for clinical measurement.The CCT was( 535±33 )μm from Pentacam with the ICC 0.994 and CV 0.062.The CCT was(537±36)pm from Sirius with the ICC 0.999 and CV 0.067.The difference value in the CCT between two methods was about 2 μm,presenting a in significant difference ( t =1.771,P＞0.05 ) and positive correlation ( r =0.985 ).The 95 ％ limit of agreement was ( - 11.64-15.65 ) μm within 1 SD of the mean value( ±34.27 pm),which was acceptable for clinical measurement.ConclusionsSirius and Pentacam show good agreement in the measurement of ACD and CCT.The two methods offer an alternative choice for the biological measurement of the anterior ocular segment.

Background Axial length and anterior chamber depth are important parameters for the calculation of diopter of intraocular lens ( IOL ). Objective This study was to investigate and compare the measuring outcomes of axial length and anterior chamber depth with IOLMaster,Axis- Ⅱ A-scan and ODM 1000A sonograph.Methods This a observational study.Axial length and anterior chamber depth were measured in 83 eyes of 48 patients with IOLMaster,Axis-Ⅱ A-scan and ODM 1000A sonograph by the same operator.The measuring results were compared among the three methods.Results The axial length were(25.79±0.85) mm,(25.72± 0.82 )mm and ( 26.00 ±0.83 )mm respectively with Axis- Ⅱ,ODM 1000A sonograph and IOLMaster.The difference between Axis-Ⅱ and DM 1000A sonograph was (0.07 ± 0.35 )mm without statistical difference between them (t=1.711,P =0.091 ).The difference of axial length between IOLMaster and DM 1000A sonograph was ( 0.27 ±0.29) mm with a statistical difference between them ( t =-8.570,P =0.000 ).The difference between IOLMaster and Axis- Ⅱ was (0.21 ±0.32 ) mm and showed a statistical difference ( t =- 5.931,P ＜ 0.01 ).The positive correlations were found in the axial length values by the each other comparison among the three instruments( r=0.916,0.938,0.928,P＜0.01 ).The anterior chamber depth values were ( 3.81 ±0.21 ) mm,( 3.84 ±0.25 ) mm and ( 3.83 ±0.18 )mm respectively with Axis-Ⅱ,0DM 1000A sonograph and IOLMaster.The difference of anterior chamber depth between Axis- Ⅱ and DM 1000A was (0.03 ±0.17 ) mm without statistical difference between them ( t =- 1.324,P =0.189 ).The difference in the anterior chamber depth between IOLMaster and DM 1000A was (0.01 ±0.15 ) mm and that between IOLMaster and Axis-Ⅱ was( 0.01 ±0.12)mm without any statistical differences among them (t =0.815,P=0.417 ;t=-0.900,P=0.371 ).The high correlation between anterior chamber depth measurements were found by the each other comparison in the three instruments ( r =0.735,0.813,0.823,P ＜ 0.01 ).Conclusions ODM 1000A sonograph can provide precise axial length and anterior chamber depth values.However,ODM 1000Asonograph can not substitute for IOLMaster in the measurement of the anterior chamber depth and axial length.

Background Clinical research showed that the corneal endothelial cell density value from different corneal specula microscopies exist diversity.The relevant literature of SP02,Tomey EM-3000 and SP3000P is still seldom up to now. Objective This research was to assess the repeatability of endothelial cell density measurements by SP02,Tomey EM-3000 and SP3000P respectively and the agreement among 3 kinds of endothelial microscopes.MethodsFifty-four healthy volunteers with the age 17-38 years old were included this research.The written informed consent was obtained from each subject before examination.The corneal endothelial cell densities in the right eyes were analyzed with SP02,Tomey EM-3000 and SP3000P respectively for 3 times under the automatic mode,and the analytical procedure of SP3000P measurement were divided into automatic mode SP3000P (A) and manual correction modes SP3000P( M).The repeatability of each specula microscopy was analyzed by calculating the intraclass correlation coefficients (ICC) and coefficient of variation ( CV ),and the 95％ confidence intervals and plotting Bland-Altman graphs were used to analyze the agreement among these methods.ResultsThe mean corneal endothelial cell densities in the population ＜24 years were significantly higher than the ones ≥ 24 years (t =3.692,P＜0.05 ),but no statistical difference was found between different gender ( t =0.335,P =0.739 ).The mean corneal endothelial cell densities were ( 3058 ± 260 ),( 2954 ± 229 ),( 2668 ± 258 ),( 2734 ± 268 ) cell/mm2 ; the ICCs were 0.957,0.940,0.972 and 0.972 and the CV were 0.063,0.061,0.056,0.058 for SP02,Tomey EM-3000,SP3000P (A) and SP3000P ( M ) respectively.The 95％ confidence intervals were ( - 100.8 - 306.8 ),( 162.6 - 617.4 ),( 109.9-494.1 ) and ( -0.6 - 132.6 ) cell/mm2 for between SP02 and Tomey EM-3000,SP3000P ( A ) and SP02,SP3000P(A) and Tomey EM-3000,SP3000P(A) and SP 3000P(M) respectively.ConclusionsSP02,Tomey EM-3000 and SP3000P(A) have good repeatability in the measurement of corneal endothelial cell density,however the outcome is different.Therefore,it is not interchangeable for the detection of corneal endothelial cell density.The differences of corneal endothelial cell density obtained from these instruments shall be paid high attention for their differences.SP3000P(A) and SP3000P(M) can be used interehangeably and SP3000P(A) is a preferable choice due to its convenience and quickness.

Background Central corneal thickness (CCT) is one of important parameters of the anterior eye segment.It plays a very important part in corneal refractive surgery and diagnosis of glaucoma.Contacted A-scan remains the gold standard for CCT measurement.Ophthalmologists are trying to look for a more convenient and noncontacted instrument to take place of contacted A-scan for CCT measurement.Objective This system analysis was to evaluate the difference between Pentacam and A-scan in CCT measurement.Methods A systematic literature retrieval was conducted from the MEDLINE,EMbase,Google Scholar,CBM disc and CNKI database with the limitation of publishing time from January 2005 to May 2011.The literature text was limited to the comparison of the CCT values measured by Pentacam and A-scan.The statistical analysis was performed using RevMan 5.1.0 software.Sensitive analysis was carried out and the publishing bias was analyzed using inverted funnel plot.Results A total of 26 studies met the requirement were included in this Meta-analysis with the 12 pieces of Chinese article and 14 pieces of English article,with the total eyes 3677.Heterogeneity was found anmong included studies (P =0.0003,I2 =56％) and random effects model was used.The differential value of CCT derived by Pentacam and A-scan was 1.74 μm,and no significant difference was found between Pentacam and A-scan (WMD =1.74,95％ CI:-0.69-4.16,P＞0.05).Fixed effects model was used to exclude the studies with the sample more than 100 eyes as the sensitive analysis.When fixed effect model was used,CCT by Pentacam was 2.73 μm more than A-scan,showing an insignificantly clinical difference.When studies with a sample more than 100 eyes were excluded,the CCT value by Pentacam was 2.64 μm more than A-scan,without clinically significant difference between them.No obvious publishing bias was seen in the included literature.Conclusions The difference between Pentacam and A-scan in CCT measurement is less and could be ignored.

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.