An observational study of autologous bone marrow-derived stem cells transplantation in seven patients with nervous system diseases: a 2-year follow-up.

Currently, autologous bone marrow-derived stem cell is one of the most innovative areas of stem cells research. Previous studies on animal models of nervous system diseases have shown that these cells have a good effect on nervous system disorders. The alternative treatment with stem cells for the nervous system diseases has also gradually reached to clinical application stage. The prospect is captivating, but the safety and efficacy of this procedure need further research. To observe the clinical efficacy and side effects of the treatment for autologous mesenchymal stem cells and neural stem/progenitor cells which are in differentiated form by inducing with cerebrospinal fluid in the patients with nervous system diseases, thirty patients were selected from our hospital (2009-10 to 2012-07) and were followed at 1 month, 3 months, 6 months, 1 year and 2 years after the treatment with autologous mesenchymal stem cells and neural stem/progenitor cells in differentiated form was introduced. In this paper, we will introduce the process to make cells accessible for the clinical application by the description of the changes observed in 7 cases were followed for 2 years. The time for bone marrow mesenchymal stem cells could be available for clinical needs is as early as 5 days, not later than 10 days, and the median time is 8 days, while neural stem/progenitor cells in differentiated form can be available for clinical needs in as early as 12 days, not later than 15 days, and the median time is 13.5 days (statistical explanation: Case 5 only uses autologous mesenchymal stem cells, and Case 7 has two times bone marrow punctures). The neurological function of the patients was improved in 1-month follow-up, and the patients have a better discontinuous trend (statistical explanation: sometimes the neurological function of the patients between two adjacent follow-ups does not change significantly). After transplantation, four patients appeared to have transient fever, but it was easily controlled by symptomatic treatment. Seven patients did not appear to show secondary tumor induced by transplantation of stem cells in 2-year follow-up. Thus, it suggests that the use of autologous bone marrow-derived stem cells transplantation in patients with nervous system diseases is a feasible, convenient, safe, and effective method.

Induction of human bone marrow mesenchymal stem cells differentiation into neural-like cells using cerebrospinal fluid.

To optimize a technique that induces bone marrow mesenchymal stem cells (BMSCs) to differentiation into neural-like cells, using cerebrospinal fluid (CSF) from the patient. In vitro, CSF (Group A) and the cell growth factors EGF and bFGF (Group B) were used to induce BMSCs to differentiate into neural-like cells. Post-induction, presence of neural-like cells was confirmed through the use of light and immunofluorescence microscopy. BMSCs can be induced to differentiate into neural-like cells. The presence of neural-like cells was confirmed via morphological characteristics, phenotype, and biological properties. Induction using CSF can shorten the production time of neural-like cells and the quantity is significantly higher than that obtained by induction with growth factor (P < 0.01). The two induction methods can induce BMSCs to differentiate into neural-like cells. Using CSF induction, 30 ml bone marrow can produce a sufficient number of neural-like cells that totally meet the requirements for clinical treatment.

[Effects of WWOX gene transfection on cell growth of epithelial ovarian cancer].

OBJECTIVE: To study the effects of anti-oncogene WWOX on cell growth of epithelial ovarian cancer,in order to find a new approach of gene therapy for ovarian cancer. METHODS: A eukaryotic expression vector containing WWOX was transfected into ovarian cancer cell line HO8910 in vitro (recombinant plasmid group), and positive cell clones were selected and amplified. Expression of WWOX protein was detected by western blot. Untransfected cell (blank contrast group) and transfected empty plasmid cell (empty plasmid group) were served as control groups. In vitro, the biology effect of WWOX on HO8910 cell was analyzed through the methyl thiazolyl tetrazolium test, transwell chamber cell invasion assay in vitro, agarose clony-formation and flow cytometry. In vivo, the cell of transfection was transplanted intraperitoneally in to BALB/c nude mice. The survival time and growth ability of nude mice were observed. RESULTS: (1) Recombinant plasmid group cell could steadily express WWOX protein, while in empty plasmid group and blank control group the expression of WWOX protein were not detected. (2) The growth rate of recombinant plasmid group cell was inhibited. (3) The agarose clony-formation rate of recombinant plasmid group (19.8%) was significantly lower than that of the empty plasmid group (54.5%) and blank control group (56.0%, P < 0.05). (4) Flow cytometry showed that (72.08 +/- 0. 39)% of cells was arrested at G0/G1 stage in recombinant plasmid group, while in empty plasmid group and blank control group G0/G1 stage cells were at (41.02 +/- 1.08)% and (39.31 +/- 0.67)% (P < 0.05). (5) In vitro invasion assay showed that invasion cell number in recombinant plasmid group (89.7 +/- 3.1) was not significantly different from that of empty plasmid group (91.2 +/- 1.3) and blank control group (91.4 +/- 1.3, P > 0.05). (6) In vivo test in nude mice showed that WWOX gene could inhibit tumor growth of the HO8910 cells. CONCLUSIONS: Tumor suppressor gene WWOX could interfere with the cell cycles of ovarian cancer cell and inhibit cell proliferation. As a new valuable tool, it promises to have application in the gene therapy of ovarian cancer.

Hepatitis C virus core proteins derived from different quasispecies of genotype 1b inhibit the growth of Chang liver cells.

AIM: To investigate the influence of different quasispecies of hepatitis C virus (HCV) genotype 1b core protein on growth of Chang liver cells. METHODS: Three eukaryotic expression plasmids (pEGFP-N1/core) that contained different quasispecies truncated core proteins of HCV genotype 1b were constructed. These were derived from tumor (T) and non-tumor (NT) tissues of a patient infected with HCV and C191 (HCV-J6). The core protein expression plasmids were transiently transfected into Chang liver cells. At different times, the cell cycle and apoptosis was assayed by flow cytometry, and cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) assay. RESULTS: The proportion of S-phase Chang liver cells transfected with pEGFP-N1/core was significantly lower than that of cells transfected with blank plasmid at three different times after transfection (all P < 0.05). The proliferation ratio of cells transfected with pEGFP-N1/core was significantly lower than that of cells transfected with blank plasmid. Among three different quasispecies, T, NT and C191 core expression cells, there was no significant difference in the proportion of S- and G0/G1-phase cells. The percentage of apoptotic cells was highest for T (T > NT > C191), and apoptosis was increased in cells transfected with pEGFP-N1/core as the transfection time increased (72 h > 48 h > 24 h). CONCLUSION: These results suggest that HCV genotype 1b core protein induces apoptosis, and inhibits cell-cycle progression and proliferation of Chang liver cells. Different quasispecies core proteins of HCV genotype 1b might have some differences in the pathogenesis of HCV persistent infection and hepatocellular carcinoma.

Ginkgo biloba extract suppresses hypertrophy and extracellular matrix accumulation in rat mesangial cells.

AIM: To observe the effects of Ginkgo biloba extract (EGb) on the hypertrophy of mesangial cells and the accumulation of extracellular matrix (ECM) in mesangial cells. METHODS: Cultured mesangial cells were allotted into 7 groups: normal group, solvent control group, high glucose group, low dose of EGb group, moderate dose of EGb group, high dose of EGb group, and captopril group. Activities of cell antioxidases, S phase percentage and G(0)/G(1) phase percentage, collagen IV and laminin, Smad2/3 and Smad7, TGF-beta(1) mRNA were measured by different methods. RESULTS: For EGb-treated groups, when compared with high glucose group, the cell percentage of S phase was raised and the percentage of G(0)/G(1) was lowered. The intensity of oxidative stress was weakened. The expression of Smad2/3 was greatly decreased and Smad7 was increased. Collagen IV, laminin and TGF- beta(1)mRNA were also reduced. CONCLUSION: EGb can suppress cell hypertrophy and the accumulation of ECM in rat mesangial cells, which means it could play a vital role in the delay of glomerulosclerosis in diabetic nephropathy.