1.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.
2.Epidemiologic study of chronic kidney disease among Uygur adults in rural area of Moyu county in Xinjiang
Dilimulati ; Zeng-lu LI ; Jian LIU ; Aizezhi ; Aniwa ; Zhen LIU ; Fang WANG ; Hai-yan WANG
Chinese Journal of Nephrology 2009;25(8):607-612
Objective To explore the method to carry out epidemiological investigation of chronic kidney disease (CKD) in rural area surrounding the Tarim Basin, and to elucidate the prevalence and risk factors associated with CKD among the Uygur adults in Moyu county. Methods A total of 1650 residents (age >18 years) from 15 villages in 3 rural town of Moyu county were randomly selected by using a stratified, multistage sampling. All the residents were interviewed and received physical examination and tested for random spot urine of albumin to creatinine ratio (ACR). Estimated glomerular filtration rate (eGFR) was calculated by modified factors of CKD were examined. Results Valid data of 1552 subjects were enrolled in the study. After the adjustment of age and gender component, the prevalence of albuminuria and reduced eGFR was 4.5% (95% CI 4.4-4.6) and 1.4% (95% CI 1.4-1.5) respectively. Approximately 5.4% subjects had at least one indicator of kidney damage. Age and hypertension were independently associated with CKD. Conclusions Experience and method of epidemiology investigation of CKD in the rural areas of Uygur are obtained through this study. The prevalence of CKD is 5.4% and the awareness is 12.5% in the Uygur adults of Moyu county. Independent risk factors associated with CKD are hypertension and age.
3.Tiamcinolone acetonide and indocyanine green-assisted vitrectomy combined with inner limiting membrane peeling for idiopathic macular hole
Bo, JIN ; Xue-min, JIN ; Hai-yan, ZHU ; Peng-yi, ZHOU ; Xian-guo, ZENG
Chinese Journal of Experimental Ophthalmology 2012;30(3):239-241
BackgroundWhether the peeling of the inner limiting membrane (ILMP) increase the closure rate of idiopathic macular hole is still in controversy.Some ophthalmologist recommend vitrectomy combined with inner limiting membrane peeling for the treatment of idiopathic macular hole.However,the removal of ILMP is difficult because of its similar appearance to adjacent tissues.Objective This study was to investigate the efficacy of triamcinolone acetonide(TA) and indocyanine green(ICG) double staining-assisted vitrectomy combined with ILM peeling during the surgery.Methods A consecutive case- observational study was designed.The standardized vitrectomy was performed in 25 eye of 23 cases with IMH.During the vitrectomy,TA and ICG were injected into posterior pole vitreous to visualize and assist the ILM peeling.The dying effectiveness was observed,and the closure rate of macular hole,visual acuity,intraocular pressure and complications were evaluated after surgery.Written informed consent was obtained from each patient prior to operation.Results Posterior vitreous cortex and ILM were visible and the residual vitreous and cortex were removed clearly after dying of TA and ICG in all the 25 eyes.During the following-up duration of 3-8 months,the completely anatomical reattachment of the macular area was in 22 eyes ( 88.0% ) and partially reattachment in 3 eyes( 12.0% ).The best corrected vision was 0.07-0.60 in all of the operated eyes 2 months after surgery.Conclusions TA and ICG- assisted vitrectomy combined with ILM peeling appears to be a safe and effective method for IMH repair.
4.The Microorganism Constitutes Analysis of Soy Sauce Liquor from High-Salt-Level Watery State Fermentation
Xiao-Bao XIE ; You-Sheng OUYANG ; Hai-Yan ZENG ; Chun-Hua WANG ; Yi-Ben CHEN ;
Microbiology 1992;0(03):-
A system analysis about the microbial flora of normal and abnormal soybean sauce liquor from the high-salt-level watery state fermentation was made and the dominant bacteria and yeasts were identified.On the other hand,a discussion of effect of temperature on microbial flora was made.The results indicated that there were no obvious differences about the count of aerobe,spore-producing bacteria,enterobacteriaceae,lactic acid bacteria and anaerobe between the normal and abnormal soybean sauce liquor and there were marked differences about the count of yeasts and salt-tolerant bacteria.The predominant yeasts in normal soybean sauce were Torulopsis and Saccharomyces,accounting for 55.9% and 35.3% of the total yeasts separately,and in abnormal soybean sauce were Pichia,candida and Saccharomyces,accounting for 62.8%,17.9% and 9.0% respectively.
5.The Developments of Microbial Resistance to Industrial Antiseptics and Disinfectants
Chun-Hua WANG ; Xiao-Bao XIE ; Hai-Yan ZENG ; You-Sheng OUYANG ; Yi-Ben CHEN ;
Microbiology 1992;0(04):-
These years with industrial antiseptics and disinfectants widely used, especially unscientifically used, the resistance of microorganisms is more and more serious, which brought much more difficulties to industrial producing. The developments on the resistance of microorganisms to industrial antiseptics and disinfectants, their resistant mechanisms and the control strategies are reviewed, which aids us to reasonably use industrial organic antiseptics and disinfectants in existence and provides academic and scientific basis.
6.Study on Flavonoids Producing and Kinetics in Cell Suspension Culture of Eucommia ulmoides Oliv.
Ri-Ming YAN ; Zhi-Bin ZHANG ; Xiao-Fang QIU ; Qing-Gui ZENG ; Hai YOU ; Du ZHU ;
China Biotechnology 2006;0(10):-
The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment,and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied.The results showed that B5 medium supplemented with 0.5mg/L NAA,0.6mg/L 6-BA and 30g/L sucrose,at initial pH 5.0~5.5,20g(FW)/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E.ulmoides suspension cells growth and flavonoids synthesis.The results of metabolic kinetics analysis for E.ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth,sucrose consumption and flavonoids production during the process.The maximum specific growth rate(?m),the actual growth yield based on sucrose(YG) and maintenance coefficient(m) were 0.417/d,0.619g/g and 0.0206g/(g?d-1) respectively.All these outcomes could give a basis for establishing the suspension cell culture of E.ulmoides and production of the natural active components in large-scale.
7.Development of a Multiplex PCR-Microarray Method for Detection of Important Enteropathogen
Yuan-Hai YOU ; Xun ZENG ; Wei GUO ; Yan YIN ; Mao-Jun ZHANG ; Jian-Zhong ZHANG ;
China Biotechnology 2006;0(12):-
Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.
8.Problems and Countermeasures of Research-type Key Lab Management in Higher Schools
Xing-Chun ZHAO ; Zeng-Huan ZHOU ; Sheng-Liang WANG ; Hai-Yan LI ;
Chinese Journal of Medical Education Research 2006;0(07):-
This paper investigates the main problems existing in research-type key lab construction in higher schools.Based on the investigation,countermeasures are proposed also.
9.Construction and immunological responses of recombinant adenovirus containing Epstein-Barr nuclear antigen 1 in mice.
Yan-Yan TONG ; Hong-Xia LI ; Li-Xia ZHANG ; Zhan WANG ; Ling ZHOU ; Yi ZENG ; Hai-Jun DU
Chinese Journal of Virology 2014;30(4):429-435
This study aimed to construct recombinant adenovirus expressing Epstein-Barr nuclear antigen 1 (EBNA1) against nasopharyngeal carcinoma (NPC). The C-terminal region fragment of the ebna1 gene of Epstein-Barr virus was amplified from the standard strain B95-8 by polymerase chain reaction (PCR). The gene fragment was inserted into the pDC316 shuttle plasmid using the EcoRI and BgIII restriction enzyme sites. The pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells after sequencing. The soluble protein was extracted from HEK293 cells, which caused apparent cytopathic effects. The transcription and expression of the ebna1 gene were confirmed using flow cytometry and Western blotting. rAd-ebna1 titers were measured by the TCID50. rAd-ebna1 was injected into BALB/c mice at a dose of 2 x 10(8) VP per mouse, EBNA1 epitope-specific responses were measured at 1st, 2nd, 4th and 8th weeks post-immunization. The target fragment of ebna1 (939 bp) was obtained by PCR, and was in consensus with the sequence from the standard strain B95-8. Cytopathic effects were observed after the pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells. rAd-ebna1 was successfully recombined in HEK293 cells. EBNA1 protein was detected in HEK293 cells, rAd-ebna1 titers reached 10(8) TCID50/mL. Specific responses to CD4+ epitopes of EBNA1 were detected in the immunized mice. In conclusion, rAd-ebna1 was successfully constructed and induced specific responses to CD4+ epitopes of EBNA1 in immunized mice.
Adenoviridae
;
genetics
;
immunology
;
Animals
;
CD4-Positive T-Lymphocytes
;
immunology
;
virology
;
Epstein-Barr Virus Infections
;
immunology
;
prevention & control
;
virology
;
Epstein-Barr Virus Nuclear Antigens
;
administration & dosage
;
genetics
;
immunology
;
Genetic Vectors
;
genetics
;
immunology
;
Herpesvirus 4, Human
;
genetics
;
immunology
;
Humans
;
Male
;
Mice
;
Mice, Inbred BALB C
;
Viral Proteins
;
administration & dosage
;
genetics
;
immunology
10.The function and mechanism of TH 17 cells in aGVHD early diffuse lung injury
Hai CHENG ; Jing TIAN ; Bin PAN ; Guoliang SONG ; Wei CHEN ; Zhiling YAN ; Zhenyu LI ; Lingyu ZENG ; Kailin XU
Chinese Journal of Organ Transplantation 2011;32(12):752-755
Objective To assess the functional role of TH 17 cells in allogeneic hematopoietic stem cell transplantation (allyHSCT).Methods Bone marrow monocytes and splenic T cells were enriched from C57/BL6 donors.Recipient Balb/c mice were irradiated with 7.5 Gy total body irradiation (TBI) and injected with 5 105 splenic T cells and 5 106 bone marrow monocytes.Survival was monitored daily,clinical graft-versus-host disease (GVHD) was assayed three times a week,and detailed histopathologic analyses of lung were performed at day six after Allo-HSCT.Flow cytometry analysis was performed using CD3-FITC,CD4-PE,CD45-PerCP-CyS.5 monoclonal antibodies.Cells were stained for intracellular cytokines using mouse TH 1/TH2/TH 17 cytokine kit.Results All the experimental animals showed GVHD manifestations on the day 6 after transplantation.Animals from BMT and HF groups were scarified and histological analysis of lung was performed.Absence of TH 17 cells induced severe pathologic pulmonary lesions.The histopathology of the lung tissue was characterized by disorganization,epithelia cell damage,interstitial fibroplasias,and monocytes infiltration.The proportion of TH1 and TH 17 in BMT group was (5.53 ± 0.11 ) % and ( 1.04 ± 0.34)% respectively,both significantly different from that in HF group.The levels of IL-17A and IFN-γin BMT group were (2.81 ±0.19) and (42.97 ± 0.23) pg/mL respectively.IL-17A could not be detected in HF group,yet the level of IFN-y was only (9.89 ± 0.51 ) pg/mL.IL-10 in both HF and BMT groups was not detectable.Conclusion Lung is on target of aGVHD.IL-17A may play a key role in the lung injury after transplantation.