4.A comparative study on rabbit bone mesenchymal stem cells and human amniotic epithelial cells transplantation for rabbit limbal stem cell deficiency
Jian-Min, LU ; Xiu-Li, LV ; Xiang, MA
Chinese Journal of Experimental Ophthalmology 2011;29(9):786-792
BackgroundLimbal stem cell deficiency usually leads to blindness, and traditional therapy is limited. Recent research demonstrated that bone mesenchymal stem cells ( BMSCs ) and human amniotic epithelial cells(AECs) could differentiate into many kinds of cells including corneal epithelial cells, but the outcome and effect of these cells on corneal stem cell deficiency are still unclear. ObjectiveThis study aimed to observe and compare the effects of rabbit BMSCs and human AECs transplantation for rabbit limbal stem cell deficiency. Methods Eighteen clean New Zealand rabbits were randomly divided into the amniotic stroma(AS) group, rabbit BMSCs group and human AECs group with 6 rabbits for each group. Limbal stem cell deficiency models were established by putting a piece of filter paper that had been soaked in a NaOH solution at the corneal center. Rabbit BMSCs were isolated and purified by density gradient centrifugation combined with the attachment culture method, and human AECs were collected by a sequential trypsin digestion technique,and the third generation rabbit BMSCs and the first generation human AECs were identified with RT-PCR. After that,cells were inoculated onto the denuded AS and grafted to the corneal surface of the experimental animals. Twenty-eight days after cell transplantation, the therapeutic effects were evaluated based on the corneal neovascularization and opacity scores. Corneal histopathological examination and immunohistochemistry were performed to evaluate and compare the effectiveness among AS,rabbit BMSCs and human AECs on corneal stem cell deficiency. The procedure complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThe third generation of rabbit BMSCs grew well after 12 hours, and the first generation of human AECs formed a membrane-like monolayer after 48 hours of incubation on AS. Immunohistochemistry staining showed that, 28 days after transplantation, the surface cells of the cornea were positive for cytokeratin 3 in both the rabbit BMSCs group and human AECs group.Compared with the AS group,the corneal neovascularization and opacity grades were significantly decreased in the rabbit BMSCs group( Z=-2. 983, P =0. 003 ; Z =-2. 844, P =0. 004 ) and human AECs group ( Z =-2. 817, P =0. 005 ; Z =-2.041, P =0. 041 ). Histopathological analysis exhibited that stratified corneal epithelial-like cells formed on the corneal stroms 28 days after grafting and no signs of goblet cells and neovascularization were found. Less inflammatory cells and regular collagen fiber could be seen in the rabbit BMSCs group and human AECs group. In addition,clinical observation also revealed that the corneas were much clearer in the rabbit BMSCs group than the human A ECs group( Z =-2. 091 , P=0. 037 ), but the corneal neovascularization score was similar between them (Z = -0. 267,P=-0. 789). ConclusionsRabbit BMSCs and human AECs can differentiate into corneal epithelial-like cells onthedamagedcornealsurfaceandfurtherdemonstrateremarkableinhibitoryeffectsoncornealneovascularization and inflammatory cells. The more dominant and prominent effect is the role played by rabbit BMSCsin the improvement of corneal transparency.
5.Key activities involved in conduct of clinical trials
Ai-Jian LI ; Ning-Ning XIONG ; Xiu-Qin WANG ;
Chinese Journal of Clinical Pharmacology and Therapeutics 2000;0(03):-
Key trial activities include: development of the trial protocol;development of standard operating procedures;development of support systems and tools;generation and approval of trial information documents;selection of trial sites and the selection of properly qualified,trained,and experienced investigators and study personnel;ethics committee review and approval of the protocol;review and approval by applicable regulatory authorities;enrollment of subjects into the study: recruitment,eligibility,and informed consent;the investigational product(s): quality,handling,and accounting;trial data acquisition: conducting the trial;trial data acquisition: conducting the trial; safety management and reporting;monitoring the trial;managing trial data;quality assurance of the trial performance and data;reporting the trial.
6.ZO-1 gene methylation status and its clinical significance in children with non-Hodgkin lymphoma.
Yu-Qiao DIAO ; Fan QU ; Ming-Juan YANG ; Jian-Hui MENG ; Xiu-Li ZHU ; Jian CHEN
Chinese Journal of Contemporary Pediatrics 2014;16(6):619-623
OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.
METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.
RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).
CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.
Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics
7.Analysis of iodine nutrition of people and serum thyroid hormone levels of women of childbearing age in pasturing areas of Tibet in 2009
Dan, DU ; Jian-tao, LI ; Su-mei, LI ; Xiu-wei, LI ; Hai-yan, WANG
Chinese Journal of Endemiology 2011;30(5):535-538
Objective To explore the iodine nutrition level of people, prevalence of iodine deficiency disorders and the thyroid function of women of childbearing age in pasturing areas of Tibet. Methods Thirty families were selected respectively in pastoral Dangxiong county and agricultural Qushui county of Lhasa in 2009,drinking water and edible salt samples were collected to test iodine content; at least 50 people from each crowd of the following populations including children aged 8 - 10, women of childbearing age of 18 - 49 old and male adults aged 18 - 60 were randomly sampled and to measure their urinary iodine content and for thyroid palpation. Direct titrimetric method was used to test salt iodine(GB/T 13025.7-1999); As3+-Ce4+ oxidation reduction process to test water iodine (GB/T 5750.1-2006); As3+-Ce4+ catalytic spectrophotometry using ammonium persulfate digestion to test urine iodine(WS/T 107-2006), and goiter examination was based on Diagnostic and Classificatory Criteria of Endemic Goiter (WS 276-2007). Results The median of water iodine was 1.3 μg/L in pasturing area and 0.7 μg/L in agricultural areas, there was no statistical significant difference between them(Z =- 1.809, P > 0.05).There was no iodized salt used in pastoral people, but iodized salt coverage rate was 90.0%(27/30) in agricultural residents. The median of urinary iodine among people of pasturing areas was 50.2 μg/L, lower than that of agricultural areas( 193.2 μg/L, Z =- 10.48, P < 0.01 ). However, the goiter rate in pasturing area[1.0%(1/100)]was significantly lower than that of agricultural areas[18.0%(18/100) , x2 =16.8, P < 0.01]. Serum level of FT4 and TT4 in pastoral population[(14.0 ± 2.0)pmol/L, (85.6 ± 17.5)nmol/L] was significantly lower than that of agricultural areas[(16.2 ± 6.3)pmol/L, (95.4 ± 21.1)nmoL/L, t =- 2.06, - 2.20, all P < 0.05]. The thyroid dysfunction rate[5.9% (2/34)]and subclinical hypothyroidism rate[2.9% (1/34)]in pastoral population was significantly lower than that of agricultural areas[25.5%(12/47), 21.3%(10/47), x2 =5.328, 5.651, all P < 0.05]. Conclusions Pastoral areas of iodine intake is significantly lower than the agricultural areas, urinary iodine levels reflect a serious iodine deficiency in pastoral people, but the blood biochemical and urinary iodine and goiter rate does not match,and shows hidden iodine hunger, which does not constitute a goiter epidemic.
8.Study on Cellulose Synthase Gene of Chinese White Poplar:Cloning,Sequencing and Construction of Higher Plant Expression Vector
Chun-Xiu LI ; Li-Wang QI ; Jian-Hua WANG ; Shou-Gong ZHANG ;
China Biotechnology 2006;0(02):-
The DNA fragment was cloned from the cDNA that was synthesized using the RNA from Chinese white poplar and verified by sequencing This cDNA clone ,designated PtoCesA1, was 3215 bp long with an opening reading frame of 2934bp extending from nucleotides 52~2985.Comprision of the nucleotide sequence of PtoCesA1 with PtrCesA1 showed 97% identity.For construction of the PtoCesA1 binary expression vector, the BamHI site was made by synonymy mutation, the full length PtoCesA1 cDNA was subcloned into pBI121 between the BamHI site and XbaI site. The PtoCesA1 binary vector,containing PtoCesA1,was verified by digestion and PCR.
9.Molecular typing on Leptospira interrogans isolates from Jiangxi province,by pulsed-field gel electrophoresis
Jian-Min XU ; Xiu-Gao JIANG ; Xiu-Wen LI ; Yan ZHANG ; Jian WANG ; Chang-Hui XIONG
Chinese Journal of Epidemiology 2010;31(4):434-437
Objective To perform a molecular epideminlogical investigation on the types of Leptospira interrogans isolates from leptospirosis patients and animal hosts in Jiangxi province,using a pulsed-field gel electrophoresis (PFGE).Methods The extracted chromosomal DNA from leptospiral isolates were digested with restriction endonuclease Not Ⅰ and the DNA segments were separated by using PFGE.By BiOnurerics V4.0 software and 75% similarity as the standard,the obtained PFGE images from leptospiral isolates were managed to establish a digitization database and then the PFGE maps of leptospiral isolates were compared with those of reference standard strains belonging to 15 serovars in 15 serogroups of L.interrogans,for cluster analysis.Results 139 strains of L.interrogans isolated from different areas of Jiangxi province were classified into 46 PFGE types.Among the PFGE types,LepNot Ⅰ.0071,LepNotⅠ.0072 and LepNot Ⅰ .0043 were the predominant types that accounting for 28.06%,15.11% and 7.19% of all the leptospiral isolates,respectively.The PFGE maps from 84.89% (118/139) of the 139 leptospiral isolates were found to basically match those of 6 reference standard strains belonging to 6 serovar in 6 serogroups of L.interrogans.In the 118 matched ieptospiral isolates,32.37% (45 strains),15.83% (22 strains) and 15.11% (21 strains)belonged to sero-groups Icterohaemorrhagiae serovar Lai,sero-groups Australis serovar Australis and sero-group Javanica serovar Javanica,respectively.Conclusion PFGE seemed a fast,accurate and effective method for typing of L.interrogans isolates.Serogroup Icterohaemorrhagiae serovar Lai and followed by serogroup Australis serovar Australis as well as serogroup Javanica serovar Javanica were the predominant L.interrogans species in humans and animal hosts in Jiangxi province.
10.Progress on treatment of transverse patella fractures with tension band fixation.
Xiu-ling YU ; Chao XU ; Shun-dong LI ; Jian-dong ZHAN ; Zai-qiang XU
China Journal of Orthopaedics and Traumatology 2015;28(11):1069-1074
Transverse fracture is the most common in patella fracture and tension band fixation is one of the most effective methods. Surgical wire tension band technique is simple, the use of materials is also simple, but it is not strong and difficult to promote. Kirschner tension band technique can get satisfactory reduction with reliable fixation, but it is easy to complicate with steel wire breakage and Kirschner loosening. Screw tension band technique inherits the traditional advantages of simple manipulation and reliable fixation, also overcomes the disadvantages of early activity limitations caused soft tissue irritation of tension band around knee, the slippage and breakage of internal fixation, and the technique can be popularized generally.
Bone Wires
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Fracture Fixation, Internal
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methods
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Humans
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Patella
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injuries
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surgery
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Stress, Mechanical