2.Effect of α-GalCer-activated natural killer T cell on survival of allograft with high-risk rejection after retrobubar injection
Yan, GONG ; Li-yan, SONG ; Hai-cheng, SUN
Chinese Journal of Experimental Ophthalmology 2012;30(3):209-212
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.
4.An analysis on the urinary thiocyanate of 149 health crowd in Nanjing.
Xiao-lian QIAN ; Hai-yan SONG ; Jian-ling BAI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(8):595-596
Adult
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Female
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Food Industry
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manpower
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Humans
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Male
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Middle Aged
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Smoking
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epidemiology
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Thiocyanates
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urine
6.Isolation and Identification of Porcine Circovirus Type 2 Virus from a Domesticated Wild Boar
Hai-Yan WANG ; Song GAO ; Xiu-Fan LIU ;
Microbiology 2008;0(11):-
The nucleic acid fragment of porcine circovirus type 2(PCV2)was amplified using PCR from the tissues of a domesticated wild boar with suspected postweaning multisystemic wasting syndrome(PMWS).The sample were then inoculated into the Dulac cells and passaged for 8 generation.The 4 nucleic acid fragments covered the complete genome for PCV2 were obtained by over-lapping PCR and sent to sequence.The sequence of genome was compared with other 9 strains originated from piglets in the same area.The result showed that these strains were in high homology.
7.Detection of antifungal agent activity against the filamentous fungi by Etest
Yang SONG ; Ding-Xia SHEN ; Hai-Ying SONG ; Yan-Ping LUO ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To investigate the susceptibility of amphotericin B,itroconazol and voriconazole against filamentous fungi.Methods Etest was used to determine the MIC of amphotericin B, itroconazol and voriconazole against filamentous fungi including Aspergillus,Penicillium,Alternaria alternate,Mucor and Rhizopus species.Results The average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus fumiagtus is O.29 ?g/ml,1.16 ?g/ml and 5.88 ?g/ml;the average MIC of amphotericin B and voriconazol to Aspergillus flavus is 6.39 ?g/ml and 0.22 ?g/ml;the average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus niger is 0.69,2.31,and 19.75 ?g/ ml.Most of Penicillium are susceptable to amphotericin B,but 3 strains showed very high MIC to voriconazol and itroconazol.Both of the testing strains of Mucor and Rhizopus were resistant to all of the three antifungal agents.Conclusion Amphotericin B,itroconazol and voriconazole possessed different susceptibility on different types of filamentous fungi.It is important for clinical laboratories to identify the filamentous fungi to the level of genus and species to help physicians choose antifungal agents correctly.
8.Effect of collagen crosslinking on porcine sclera with different methods
Yuan, WU ; Song-lin, YANG ; Hai-li, LI ; Xiao-ming, YAN ; Fan, SONG
Chinese Journal of Experimental Ophthalmology 2013;(2):168-171
Background Chemical crosslinking agent can be used to strengthen the intensity of sclera tissue,but the intensity of the sclera may be influenced by different crosslinking methods.Objective The aim of this study was to compare the effectiveness of collagen crosslinking on porcine sclera between whole-eye crosslinking method and scleral strip crosslinking method.Methods Whole-eye crosslinking or sclera strip crosslinking was performed on 70 fresh porcine eyeballs in five groups using 1% genipin,1% glutaraldehyde or PBS respectively for 40 minutes.After crosslinking,10 sclera strips with l0 mm×4 mm from the temporal lateral were prepared in every group for the stress-strain measurement using a Instron5848 microtester,and the other 4 scleral strips in each group were extracted for the thermal shrinkage temperature test.Results Biomechanical property test reveled that the elastic modulus value of sclera strips reduced by 70.0%-82.8% in the whole-eye crosslinking method group compared with scleral stip crosslinking method group after treated with 1% genipin ((8.98 ± 1.81) MPa vs.(10.85 ± 1.83) MPa,t =3.375,P =0.003)) and 1% glutaraldehyde((12.78 ±2.91) MPa vs.(18.25 ±5.16) MPa,t =4.007,P =0.001)) ;The tensile stress of whole-eye crosslinking method group was 54.9%-90.1% of scleral stips method group,showing significant decline after corsslinked of whole-eye in 5%,10%,15% and 20% strain conditions (all P < 0.05).Thermomechanics test showed that the thermal shrinkage temperature was lower in the whole-eye crosslinking group compared with scleral stip crosslinking group after treated with both 1% genipin ((68.8 ±0.9)℃ vs.(74.8± 1.3)℃,t=11.129,P=0.000)) and 1% glutaraldehyde((73.3±0.9)℃ vs.(79.3±1.3)℃,t=11.112,P=0.000)).Conclusions Different crosslinking methods have an influence on the efficacy of collagen crosslinking on porcine sclera.Sclera strip crosslinking offers a better crosslinking intensity for selera.
9.Effects of X-rays and γ-rays on reconstitution of hematopoiesis and immunity after allogeneic bone marrow transplantation
Bin PAN ; Lingyu ZENG ; Hai CHENG ; Guoliang SONG ; Lu JIA ; Zhiling YAN ; Chong CHEN ; Kailin XU
Chinese Journal of Radiological Medicine and Protection 2011;31(3):260-263
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.
10.Surgical treatment of severe ocular injury in ironware manufacturer.
Song CHEN ; Jing CUI ; Jun LI ; Ying-hai XU ; Hua YAN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(3):228-229
Adult
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Eye Injuries
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surgery
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Female
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Humans
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Male
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Middle Aged
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Occupational Diseases
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surgery
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Vitrectomy