1.Glutamine and pediatric nutrition.
Chinese Journal of Pediatrics 2004;42(7):544-547
3.The research progress in induced pluripotent stem cell in ophthalmology
Chinese Journal of Experimental Ophthalmology 2012;30(7):662-666
The establishment of induced pluripotent stem cells(iPSCs)has been a major breakthrough in the field of stem cell research since 2006,and it made possible for the use of stem cells in treating retinal degenerative diseases.Research showed that fibroblast,B lymphocytes,neural stem cells,hair corneous cells,pancreatic cells,mesenchymal cells of umbilical cord stroma and amniotic membrane can be reprogrammed as iPSCs,and they are capable of differentiating into specific types of cells.Some novel developments in iPSCs study in ophthalmology also were observed over the past few years.Induced iPSCs can differentiate into retinal pigment epithelial cells,photoreceptors and other retinal cells,which lay a foundation for the therapy of retinal degenerative diseases.Differented from traditional treatment of stem cells,the generation of iPSCs makes it possible to utilize somatic cells derived from patients for stem cell therapy without provoking ethical and immunological problems.The generation of iPSCs,the current research about iPSCs in the ophthalmic field,the limitations of iPSCs in the clinic and their future development and application were reviewed.
4.Experimental study of the oriented differentiation of bone marrow derived mesenchymal stem cells into chondrogenic phenotype in a specific culture fluid
Xiaofei DING ; Jingmin ZHAO ; Weiping CHEN ; Zhi YANG ; Wei SU
Chinese Journal of Tissue Engineering Research 2005;9(34):167-169
BACKGROUND:To construct tissue engineering cartilage would open up a novel way for the repair of cartilage damage in avoidance of the disadvantages of traditional therapeutic method.OBJECTIVE: To probe the techniques for the isolation of mesenchymal stem cells (MSCs) from bone marrow, as well as the in vitro differentiation into chondrocytic phenotype in a specific culture fluid.DESIGN:A complete randomized experimentSETTING:The Department of Traumatic Orthopedics and Hand Surgery of the First Affiliated Hospital of Guangxi Medical University, and Teaching and Research Faculty of Histology and Embryology of Guangxi Medical University.METHODS: The experiment was carried out at Guangxi Medical University between August 2002 and April 2003. Twenty SD neonatal weaning rats were selected. Bone marrow was aspirated from the bones of rat limbs and was isolated by gradient centrifugation in Percoll, and MSCs could be obtained in combination with adherent screening method, which were then cultured in DMEM-LG with 15% fatal bovine serum (FBS) in the incubator of 37℃ with 5% CO2 for 10-14 days. The passage cells were induced in DMEM-HG with 15% FBS (containing TGF-β1 10 μg/L, 10-7 mol/L dexamethasone, 50 mg/L VitC).MAIN OUTCOME MEASURES :The morphology, growth, as well as proliferation and specific expression of chondrogenic matrix of in vitro cultured MSCs due to specific induction.RESULTS: Totally 20 SD rats were observed and analyzed with no loss SCs grew in visible symmetric colonies, displaying a long-spindle shape,and the morphological characteristics of marrow-derived MSCs had no obvious changes during passage-culture, but its proliferation time was found from a shuttle fibroblastic appearance to polygonal shape, displaying posiHC staining of type Ⅱ collagen of cartilage specific matrix.bronectin adherent screening technique is a convenient, effective and practical method to separate and collect MSCs from rat bone marrows in chondrogenic phenotype when induced by a specific medium and can secrete cartilage specific matrix, and they can be the optimal seed cells for cartilage tissue engineering.
6.TLR3 and TLR7/8 ligands induce microglia activation in intact rat spinal cord
Yan-Hua SU ; Zhi-Ming ZHANG ; Zhi-Ren ZHANG ; Ben-Huang ZHAO
Chinese Journal of Neuromedicine 2009;8(8):764-768
Objective To study the effects of two stimulators of the innate immune system, polyinosine-polycytidylic acid [Poly (I:C)] and R848, on the activation of microglia in rat spinal cord. Methods Thirty male Lewis rats (6 to 8 weeks old) were divided into Poly(I:C) group (n=12), R848 group (n=15) and control group (n=3). According to the killing time, the Poly(I:C) group and R848 group were divided into 4 and 5 sub-groups, respectively, with 3 rats in each sub-group. The rats in the subgroups received intraperitoneal injection of a single bolus of Poly(I:C) (5 mg/kg) or R848 (1 mg/kg) accordingly, and in the control group, the same volume of phosphate-buffered saline (PBS) was administered. Activated microglia were observed using immunohistochemistry for ED-1, AIF-1, EMAP Ⅱ, OX6, and P2X4R, and BrdU staining was used to identify the proliferating cells. Results Compared with the control group, both Poly (I:C) and R848 groups showed a significant but transient increase of ED-1-positive spinal cord microglia 4 days after the injection, while no significant differences were found in the microglial markers AIF-1, EMAP Ⅱ, OX6, P2X4 receptor (P2X4R), indicating that the microglia were not fully activated. Tracing of the cell proliferation by BrdU revealed that only a small fraction of the proliferating cells were microglia (less than 5%). Conclusion Poly(I:C) and R848 have definite effects on the innate immune system of the spinal cord and modulate the immune activity in the spinal cord, suggesting the value of these agents in modulating local regenerative processes in injured spinal cord.
7.Expression of neural stem cells transfected by neurenergen-3 gene
Zhandong BO ; Jinmin ZHAO ; Zhi YANG ; Wei SU ; Guangxiang HONG ; Zhengwang LEI
Chinese Journal of Tissue Engineering Research 2008;12(12):2374-2378
BACKGROUND: It is one of hot topics for the application of neurotrophic factors including neurenergen-3 to promote peripheral neural regeneration nowadays; however, clinical application is restricted to safety and effective administration route. Gene transfection brings a novel thinking and pathway for neurotrophic factors used in clinic.OBJECTIVE: To observe the expression of neural stem cells modified by neurenergen-3 gene after transfection.DESIGN: Completely randomized study.SETTING: Department of Traumatic Orthopaedics and Hand Surgery, the First Affiliated Hospital, Guangxi Medical University;Department of Hand Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Three healthy SD rats of four months old and either gender were selected from Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Recombinant adenoviral expressing vector for transfection of neural stem cells was constructed in Laboratory of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, and the concentration was 0.15 g/L.METHODS: The experiment was carried out in the Orthopaedic Laboratory and Central Laboratory, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from December 2002 to March 2004. Recombinant plasmid pAD-neurenergen-3 containing with green fluorescent protein (GFP) gene was transfectd into primarily cultured neural stem cells by using cationic tiposome interventional method. At 72 hours after transfection, fluorescent inverted microscope was used to observe GFP expression in neural stem cells, and transfection efficiency was measured simultaneously. Expression and transcription of neurenergen-3 gene in neural stem cells were detected at 72 hours, 1 and 5 weeks after transfection by using immunocytochemical stain and reverse transcription polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of neurenergen-3 gene in transfected neural stem cells.RESULTS: ① Neural stem cells transfected by recombinant plasmid pAD-neurenergen-3: GFP expressed on partial neural stem cells at 72 hours after transfection, and the transfection efficiency was 40%. Five weeks later, GFP expression was still observed. ②Transcription and expression of neurenergen-3 gene in neural stem cells: Transcription of neurenergen-3 mRNA was observed in neural stem cells at 72 hours, 1 and 5 weeks after transfection, and expression of neurenergen-3 mRNA was still observed in 5 weeks after trans fection.CONCLUSION: As the carrier of cationic liposome, neurenergen-3 gene can effectively transfect, culture and long-term express neural stem cells via the introduction of adenoviruses.
9.The clinical significance of the blood LDH,?_2-MG,D-dimer measuring in the diagnosis and treatment of non-Hodgkin lymphoma
Su-Ying QIAN ; Zhao-Xiong XIA ; Jian-Fen XU ; Ya-Min CHEN ; Zhi FANG ;
Cancer Research and Clinic 2006;0(09):-
Objective To explore the clinical significance of the blood lactic dehydrogenase(LDH), ?_2-microglobulin(?_2-MG),D-dimer measuring in the diagnosis and treatment of Non-Hodgkin lymphoma. Methods In 40 cases with NHL,LDH was measured by L-P continuous monitoring method,?_2-MG was measured by luminescent immunoassay,D-dimer was measured by immunoturbidimettic assay.Results The levels of the blood LDH,?_2-MG and D-dimer in patients with NHL were higher than those of in the controls(P 0.05).Con- clusion The levels of blood LDH,?_2-MG,D-dimer can be taken as an auxiliary clinical index to diagnose, classify the phase,evaluate the effectiveness of treatment and prognosis in the NHL patients,and have impor- tant clinical significance.
10.Forensic validation of goldeneye? DNA ID 26Y system.
Ting-zhi QUE ; Yuan LIN ; Zhen-min ZHAO ; Yan LIU ; Su-hua ZHANG
Journal of Forensic Medicine 2014;30(6):446-455
OBJECTIVE:
To perform the validation and analysis of forensic parameters of Goldeneye DNA ID 26Y system.
METHODS:
Based on the validation rules of Scientific Working Group on DNA Analysis Methods (SWGDAM), the kit was assessed from several parts, as test of PCR system, reproducibility, accuracy, and sensitivity, etc. And Y-STR loci of 517 unrelated healthy individuals from Eastern China were genotypes by this kit. The distribution and frequency of haplotype were calculated and forensic parameters of the kit were assessed.
RESULTS:
The complete profiles can be obtained even when the PCR reaction volume with 6.25 microL. And correct profile was obtained with DNA down to 125 pg. No reproducible peaks were detected with the DNA of common animals and microorganism with the kit. For the male-male mixture testing, average 70% of the minor alleles were obtained when the ratios of 1:19 and 19:1. For the male-female mixture testing, results showed that the sensitivity of the kit was no compromised with the addition of female samples.
CONCLUSION
The validation studies demonstrated that Goldeneye DNA ID 26Y system has good sensitivity and specificity, and suitable for mixture testing. The polymorphism of 26 Y-STR loci included in this kit are good for forensic application.
Alleles
;
Animals
;
Asian People/genetics*
;
China
;
Chromosomes, Human, Y
;
DNA
;
DNA Fingerprinting/standards*
;
Female
;
Forensic Genetics/methods*
;
Genotype
;
Humans
;
Male
;
Polymerase Chain Reaction
;
Polymorphism, Genetic
;
Reproducibility of Results
;
Sensitivity and Specificity