ABSTRACT
Objective To establish transgenic mouse models expressing human HSP22 protein.Methods pCAGGS-HA-Wt HSP22 transgenic expressing vector carrying human HSP22 gene was constructed by gene recombination technology.The linearized DNA was got by SalI、Hind Ⅲ and BsaⅪ digestion of PCAGGS-HA-Wt HSP22,purified and microinjected into fertilized eggs from C57BL mice.The tail DNA of pups was tested by PCR and DNA sequencing.Expression of human HSP22 protein was detected by western blot with anti-HA tag monoclonal antibody.Results 4 transgenic founder mice (Tg646,Tg648,Tg649,Tg661) carrying human HSP22 gene were identified by PCR and DNA sequencing.The human HSP22 protein was expressed in the lines Tg646,Tg648 and Tg649 founder mice,but was not expressed in the line Tg661 founder mouse.Conclusions The mouse models expressing human HSP22 protein are established successfully and provide the foundation for HSP22 gene research in vivo.
ABSTRACT
BACKGROUND: Basic fibroblast growth factor (bFGF) can accelerate the bone marrow mesenchymai stem cells (BMSCs) proliferation and differentiation into nerve cells, which is considered as a mitogen of glial cells.OBJECTIVE: To investigate the survival and differentiation of bFGF gene modified BMSCs transplanted on rat models of cerebral ischemia by double immunofluorescence staining, and to study the differentiation trend of BMSCs into neuron-like cells and glial cells.DESIGN, TIME AND SETTING: The randomized control animal experiment was completed in the central laboratory of Experimental Animal Center of Central South University from July 2005 to March 2006.MATERIAL: Fifty Sprague-Dawley rats were divided into 4 groups at random: sham operation group (n=10), cerebral ischemia-reperfusion injury group (n=10), BMSCs group (n=i5) and bFGF modified BMSCs group (n=15). METHODS: Except sham operation group, rats in the other three groups were prepared for local cerebral ischemia-reperfusion models. Then BMSCs or bFGF modified BMSCs were intravenously transplanted into cerebral ischemic rats, and the same volume of DMEM were injected in the cerebral ischemia-reperfusion injury group. MAIN OUTCOME MEASURES: Survival rate and differentiation of grafted cells were observed by 5-bromo-2-deoxyuridine (BrdU)-NeuN, and BrdU-glial fibrillary acidic protein (GFAP) double immunofluorescence staining; the neurological scores and infarction volumes in each group. RESULTS: At 7 days after implantation, the number of BrdU/NeuN-positive cells and BrdU-GFAP-positive cells in the bFGF modified BMSCs group was higher than those on the BMSCs group (P < 0.05), but there were no significant differences in the co-expression cells by double immunofluorescence staining between the two groups (P > 0.05). At 7 days following reperfusion, neural function of cerebral ischemia rats were improved and infarction volume was reduced in both BMSCs group and the bFGF modified BMSCs group, and bFGF modified BMSCs group was superior to BMSCs group. CONCLUSION: BMSCs modified by bFGF can survive in cerebral ischemic region and differentiate into neuron and glial cells, which are more proper for repairing nerves.
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Objective To evaluate the clinical and genetic characteristics of Charcot-Marie-Tooth disease (CMT). Methods The clinical materials and hereditary histories of 110 cases in 70 families with CMT were analyzed retrospectively.Results The ratio of male to female was 2.03∶1. The age at onset was from 1 to 61 years old and the mean age was 19.1 years old. 78.2% of the patients had CMT before 30 years old.70 patients (63.6%) had positive family history and they showed mostly autosomal dominant inheritance. The rate of consanguinity was 6.9%. In the CMT group, we could find the amyotrophy of legs in 106 patients (96.4%), distal muscle weakness and atrophy of the upper limbs in 48 patients (43.6%), stork legs in 64 patients (58.2%), pes cavus in 68 patients (61.8%), decreased or diminished tendom reflexes in 108 patients (98.2%). Electromyography examination in 61 patients showed neurogenic damages. Muscle biopsy in 37 patients showed neurogenic amyotrophy. Sural nerve biopsy was performed in 25 patients. 20 patients were charactered by demyelination, Schwann cell proliferation and/or “onion bulbs” change and 5 patients were associated with axis cylinder degeneration, but tomaculous change was not found in all the 25 patients.Conclusions In the CMT group, the onset age was mostly in childhood and adolescence. The male CMT cases were more than the females. Autosomal dominant was the mostly frequent inheretance. Neuroelectrophysiology and pathological examination are important for the diagnosis and type of CMT.