BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.

Background and purpose:To explore the possibility of genetic re-expression silenced by DNA aberrant hypermethylation which is a common epigenetic modification in carcinogenesis. 5-Aza-CdR, an inhibitor of DNA methylation, was used to determine the effects of expression of tumor suppressor gene E-cadherin in tumor cell lines. Methods:Methylation specific PCR(MSP) was utilized to examine methylation status of E-cad gene on breast carcinoma cell line MDA-MB-435 before and after the treatment with 5-Aza-CdR. Immunohistochemistry(IHC) was used to test the expression of E-cad protein. Semi-quantitative RT-PCR method was used to detect the changes of E-cad mRNA.Results:1).E-cad methylation was positive(116bp) and unmethylation was negative on MDA-MB-435 cell before the treatment with 5-Aza-CdR. After being treated with 5.0umol/L 5-Aza-CdR for 3 days, methylation turned negative and unmethylation positive bands(97bp) were detected. 2).The E-cad protein expression was not detected by immunohistochemistry on MDA-MB-435 cell before the treatment, while E-cad staining was positive on the cell membrane after the treatment. 3). The E-cad mRNA failed to be amplified in cells before the treatment. After incubation at variable concentrations of 0.5 ?mol/L, 1.0 ?mol/L, 2.0 ?mol/L and 5.0 ?mol/L 5-Aza-CdR for 3 days, respectively, E-cad mRNA expression was detected on the fourth day in a dose-dependent manner. Correlation between the mRNA expression level and the agent concentration was observed.Conclusions:The demethylation agent 5-Aza-CdR can reverse the aberrant E-cad methylation status in MDA-MB-435 and re-expressed E-cad mRNA and protein.

Objective To analysis the clinical manifestations of a large Spinocerebellar Ataxia 3 pedigree to pro-vide the information for the early diagnosis of Ataxia 3. Methods SCA3/ATXN3 gene was determined by using Poly-merase Chain Reaction and fragment analysis in the large pedigree members and patients ’clinical data was collected. Five patients underwent MRI imaging and fundus examination. Results There were eighteen clinical patients and twelve ATXN3 carriers in this Pedigree . In addition to ataxia, three patients presented with intellectual disability, one with cer-vical spondylosis, one with dysmyotonia, one with disorder in visual system, and seven with abnormality in autonomic ner-vous system. The MRI revealed that pons and cerebellar atrophy in some patients inordinately. Undus examination did not reveal any obvious abnormality. Conclusions The symptoms of SCA3 are heterogeneous in the same pedigree. When patients present with symptoms of cerebellar system, visual system and autonomic nervous system, or cervical spondylosis and intellectual disability, SCA3 should be considered.

BACKGROUND:Some studies have shown that more copy number variations are present in early passage human induced pluripotent stem cells than later passage human human induced pluripotent stem cells, their parental somatic fibroblasts or human embryonic stem cells. OBJECTIVE:To investigate whether the reprogramming process itself compromises genomic stability and further explore the efficiency of induced pluripotent stem cellestablishment. METHODS:Using high-resolution Affymetrix CytoScan HD array, we compared copy number variations and loss of heterozygosity in early passage induced pluripotent stem cells with their fibroblast cellorigins from genetic epilepsy patients. RESULTS AND CONCLUSION:Compared with somatic fibroblasts from genetic epilepsy patient, there was no difference in the loss of heterozygosity between the two types of cells, but more copy number variations were present in early passage human induced pluripotent stem cells which were characterized as microduplication and involved oncogenic genes. Results demonstrate the dynamic nature of genomic abnormalities during reprogramming process and the necessity of frequent monitoring human induced pluripotent stem cells to assure their genomic stability and clinical safety.

Objective To investigate the etiologic effect of the lumbar facet joint morphology on the occurrence of degener?ative lumbar spondylolisthesis (DLS). Methods From January 2007 to July 2013, 115 patients with DLS treated in our hospi?tal were randomly selected. There were 28 males and 87 females with an average age of 57.3 years (range, 41-76 years). 115 age? and sex? matched people including 31 males and 84 females with an average age of 56.4 years (range, 45-77 years) free from DLS and back or leg pain were selected randomly as control group from a group coming for routine physical examination in our hospital. Both groups received lumbar anteroposterior and lateral X?ray films、CT scanning and multiplanar reformation, the degree of spondylolisthesis (Taillard index) was measured in DLS group on lateral radiographs; at L3,4 and L4,5 level of both groups the facet joint angles on CT scan images were measured and facet tropism was evaluated, the pedicle?facet angle (the P?F angle) was measured in the sagittal plane on multiplanar reformation CT images, and then all angles of corresponding level were compared and analyzed; L4,5 facet joint degeneration in both groups was evaluated and compared in bone window, the de?gree of spondylolisthesis (Taillard index) in DLS group at different degenerative grade of facet joints were analyzed. The corre?lation between L4,5 facet joint angle、P?F angle and degrees of spondylolisthesis were analyzed. Results All L4,5 spondylolisthe?sis in DLS group were grade I, the facet joint angles were more sagittal in DLS group than those in the control group at L 3,4 and L4,5 levels, and the P?F angles were more horizontal in DLS group compared with control group;the facet tropism in DLS group at L 4,5 level were significantly different as compared to the control group, but there was no significant difference at L 3,4 level between the two groups. Significant difference was found in L4,5 facet joint degeneration grade between two groups,but there was no significant difference in degree of spondylolisthesis during different degeneration grades in DLS group. There was no significant correlation between the facet joint angle and the P?F angle and degree of spondylolisthesis at L4,5 level in DLS group. Conclusion The facet joint morphology abnormality (smaller facet joint angle, horizontal P?F angle, the facet tropism) has an important etiologic meaning in the occurrence of degenerative lumbar spondylolisthesis, however its role cannot be excessively exaggerated. The facet joint de?generation is a secondary change with aging,while the development of DLS aggravates the degeneration.

Objective:To compare the clinical effect of anterior and posterior resection,bone grafting and internal fixation on the spinal tuberculosis.Methods:82 cases of patients with spinal tuberculosis in our hospital from February 2014 to August 2015 were selected and randomly divided into the control group and the observation group with 41 cases in each group.Both groups received conventional anti tuberculosis treament and underwent debridement bone graft and internal fixation treatment,the control group underwent anterior internal fixation,while the observation group underwent posterior internal fixation.The operation time,intraoperative blood loss,low back pain relief time,ambulation time and hospitalization time were compared between two groups.All patients were followed up for 6 months,the Frankel grade and serum Thl7 cell related factors levels were compared between two groups before and after operation.Results:The low back pain relief time,ambulation time and hospitalization time of observation group were significantly shorter than those of the control group(P＜0.01).After operation,the Frankel classification grade A and B ratio of observation group were significantly higher than those of the control group (P＜0.05),the serum IL-10,IL-17,IL-23 and TGF-beta 1 levels were significantly lower than those of the control group (P＜0.01).Conclusion:Anterior resection combined with posterior bone graft inFixation could effectively promote the rehabilitation of spinal tuberculosis,reduce the severity of spinal injury,relieve the inflammatory response and promote the recovery of immune function.

This paper is to show a way of acqusition of the variable region gene of rabbit osteoprotegerin (OPG) and to analyse series. Total RNA was extracted from rabbit tibia, transcripted reversely into cDNA with random primers. The variable region of the OPG gene ampliflied using 5'RACE. Sequencing was confirmed by agarose gel electrophoresis and sequencing analysis. Full length of OPG gene was 1540bp that encoding 400 amino acids. It shared 89% identity with human OPG in whole amino acid sequence and about 85% with rattus norvegicus and other mammal. The OPG sequence of rabbit was obtained by 5'RACE, which could provide a good basis for OPG functional study.
Animals ; Base Sequence ; Molecular Sequence Data ; Osteoprotegerin ; genetics ; Rabbits ; Sequence Homology, Amino Acid

The finite element model of the intact lumbar spine (L1-L5) was set up to study the biomechanical changes of three different pairs of the lumbar laminectomy. The three-dimensional finite elements model of L1-L5 vertebrae structure was constructed by the combination of self-compiled software and Hyper Mesh. The finite element model was compared with the experimental data in vitro. The finite element model was modified of stenosis at L3-L4 and L4-L5 with the same boundary conditions and physical loads to study the motion and loading in the annulus changes at the surgical site as a result of surgical alteration. The study suggested that the removal of posterior lumbar spinal elements for the treatment of stenosis at L3-L4 and L4-L5 produced a graded increase in motion at the surgical site, with the greatest changes occurring in flexion-extension and axial rotation and that during lateral bending the amount of resection was only slightly affected. The data showed that for flexion-extension and axial rotation the increases in motion were correlated to the extent of posterior element removal. It is necessary to retain the greatest degree of posterior lumbar structures in thorough decompression, which can further reduce the postoperative intervertebral disc, facet degeneration.
Adult ; Biomechanical Phenomena ; Computer Simulation ; Finite Element Analysis ; Humans ; Laminectomy ; methods ; Lumbar Vertebrae ; diagnostic imaging ; surgery ; Male ; Models, Biological ; Spinal Stenosis ; diagnostic imaging ; surgery ; Tomography, Spiral Computed

Objective To investigate the current status of urinary incontinence (UI) at the third trimester of pregnancy among primiparas in Guangdong Province.Methods The convenience sampling method was adopted to conduct the retrospective analysis on 714 primiparas in 3 hospitals of Guangdong Province from June 2015 to March 2016.International Consultation on Incontinence Questionnaire Urinary Incontinence Short-Form Chinese was used to investigate the occurrence status of UI at the third trimester of pregnancy among primiparas.And its influencing factors were analyzed.Results Among 714 primiparas,192 cases (26.9％) developed UI during the third trimester of pregnancy,in which stress UI accounted for 67.2％ (129/192).The prepregnancy BMI and abortion had statistical difference between.the UI patients and non-UI patients (P＜0.05).The multivariate Logistic regression results showed that pre-pregnancy BMI was correlated with UI at the trimester stage of pregnancy (OR =1.077,P＜0.05).Conclusion Pre-pregnancy BMI might be an independent risk factor of UI occurrence at the third trimester of pregnancy.

OBJECTIVETo clarify the clinical, cytogenetical and molecular characteristics and prognosis of Ph(+) ALL patients with ABL kinase domain mutations (ABL-KDMs), and to evaluate the therapeutic value of allogeneic hematopoietic stem cell transplantation (allo-HSCT) combined with tyrosine kinase inhibitor (TKI) in these patients.

METHODSRetrospective analysis of clinical features, molecular genetic characteristics, mutation distribution and prognosis of newly diagnosed Ph(+) ALL patients with ABL-KDMs from February 2010 to August 2014 were performed, and the efficacy of treatment regimen of allo-HSCT combined with different TKIs was compared.

RESULTSOf 88 Ph(+) ALL patients during maintenance treatment stage for ABL-KDMs monitoring, mutation was detected in 42 patients with median time of 8 months from diagnosis to mutation occurrence. The median age of mutation group was 40-year-old, older than that of non-mutation group (32.5-year-old) (P=0.023). The incidence of complex chromosome abnormality of mutation group was higher than that of non-mutation group (P=0.043), with alternations in chromosome 7, 5 and +Ph more frequently observed. There were 21 types of mutation at 18 locations detected, with T315I mutation ranking the top followed by E255K/V, Y253H/F and E459K. Mutation group featured no significant difference in complete remission (CR) rate in contrast to nonmutation group, but was remarkably lower in major molecular remission (MMR) rate than non-mutation group. The 2 year and 5 year overall survival rate of mutation group was 45.4% and 35.0% respectively, much shorter than that of non-mutation group (67.8% and 63.3%), (P=0.047). The median survival of patients with T315I and E255K/V was 19 and 10 months, significantly shorter than that of patients with other mutations. Among the 42 patients with mutations, 14 underwent allo-HSCT, and the median survival was 29 months, longer than that of patients received chemotherapy alone (17 months) (P=0.024). Fourteen allo-HSCT patients were given nilotinib or dasatinib at the time of mutation occurrence, and there was no significant difference in the overall survival in contrast to patients who continue to take imatinib.

CONCLUSIONSABL kinase domain mutations are closely related to the older age and high genomic instability in the newly diagnosed Ph(+) ALL patients. Mutation types showed diversity and complexity, which remarkably affected patients' prognosis and survival. T315I and E255K mutations account for more than half of all cases, characterized by a less favorable prognosis. Currently, allo-HSCT is the only method that has the potential of elongating life expectancy, but the utility of second-generation TKI during relapse does not necessarily have an edge on survival over imatinib.

Chromosome Aberrations ; Dasatinib ; therapeutic use ; Hematopoietic Stem Cell Transplantation ; Humans ; Imatinib Mesylate ; therapeutic use ; Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; Prognosis ; Protein Kinase Inhibitors ; therapeutic use ; Proto-Oncogene Proteins c-abl ; genetics ; Pyrimidines ; therapeutic use ; Remission Induction ; Retrospective Studies ; Survival Rate