In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Objective To investigate the minimal erythema does (MED) of normal skin to UV in Guangzhou city,and to observe its relationship to sex,age,skin type,seasons and the years lived in Guangzhou.Methods 621 healthy subjects were exposed to Solar Simulator (GS2004) and the MED was measured and observed by two professional technicians after (24±2) hours.Results The average MED value of all subjects was (1170.2±333.2) mJ/cm2.In male and female group,the average MED values were (1132.8-339.4) mJ/cm2 and (1182.1 ± 330.7) mJ/cm2,respectively,and there was no significant difference between male and female (P=0.20).The MED value in subjects aged from 30 to 50 (1014.7 ± 359.7) mJ/cm2 was significantly lower than those aged from 20 to 29 (1222.9±304.3) mJ/cm2 and over 50 years (1179.0±374.3) mJ/cm2 (P＜0.01).The MED value in skin type Ⅱ (673.53±228.3) mJ/cm2 was significantly lower than those in type Ⅲ (1224.3±254.2) mJ/cm2 and Ⅳ(1363.1±278.5) mJ/cm2(P＜0.01).There was significant difference of the MED value between different seasons (P＜0.01).The MED value in spring (969.2±355.8) mJ/cm2 was lowest,and followed by summer (969.2± 355.8) mJ/cm2.However,there was no significant difference between autumn and winter (P＞0.05).The MED value in subjects lived in Guangzhou from 5 to 10 years was significantly lower than those whose residence time was from 1 to 4 years and over 10 years,respectively.Conclusions The MED value of the subjects in Guangzhou is quite different from other cities of China,and related to age,skin types,seasons and the years lived in Guangzhou city,while there is no correlation between MED value and gender difference.

BACKGROUND:Islet and islet cel transplantation for the treatment of diabetes has achieved effect, but the research is limited dut to the shortage of islet and immune rejection. OBJECTIVE:To observe the effect of transplantation of islet-like cells that in vitro differentiated from bone marrow mesenchymal stem cells on the treatment of diabetes in rats. METHODS:The rat bone marrow mesenchymal stem cells were induced with basic fibroblast growth factors and hepatocyte growth factors, and then received immunocytochemistry staining to detect the induction. The Sprague Dawley rats received intraperitoneal injection of streptozotocin to establish the diabetes models. After modeling, the rats were randomly divided into control group and experimental group (transplanted with induced islet-like cells). The experimental group was transplanted with the induced islet-like cells through renal capsule, and the control group was transplanted with normal saline in the same dose. The blood glucose and body mass of the diabetes rats were observed after transplantation. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells could differentiate into islet-like cells after in vitro induced with basic fibroblast growth factors and hepatocyte growth factors. There was no significant change in blood glucose of the control group after transplantation (P>0.05), and the blood glucose of the rats in the experimental group was significantly decreased compared with the control group (P<0.05). The bone marrow mesenchymal stem cells can differentiate into islet-like cells after in vitro induced with the induction system containing basic fibroblast growth factors and hepatocyte growth factors, and the islet-like cells have a certain ability of insulin secretion. The transplantation of induced islet-like cells after transplanted into the diabetes rats through renal capsule can decrease the blood glucose level of the rats.

A 48-year-old man presented with a 4-day history of fever and 10-year history of papulovesicles on the face, neck, trunk and limbs which had been aggravated 10 days prior to the presentation.Skin biopsy showed a dermal infiltration of numerous small- to medium-sized atypical lymphocytes, which was mainly located around blood vessels or appendages, with the involvement of subcutaneous fat tissue and destruction of blood vessels. The infiltrating atypical cells stained positive for CD45RO, CD8, CD56, T-cell intracellular antigen-1, granzyme B, Epstein-Barr virus-encoded small nuclear RNAs (EBER), but negative for CD20, CD79a, CD3, CD4 or CD30. Cytoplasmic CD3ε was also observed in these cells. Laboratory examinations on admission revealed a progressive decrease in peripheral erythrocytes, white cells and platelets, persistent increase in serum aminotransferase and bilirubin, and decline in serum fibrinogen and hypertriglyceridemia. The B-mode ultrasound of the abdomen showed hepatosplenomegaly. Based on the above findings,the diagnosis was made as extranodal nasal type NK/T-cell lymphoma of skin complicated by hemophagocytic syndrome.

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 throush 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 agnrose 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 premises to have application in the gene therapy of ovarian cancer.

Objective To evaluate effectiveness of peripheral Mohs micrographic surgery for the treatment of extramammary Paget′s disease (EMPD). Methods A total of 28 patients with EMPD were treated with peripheral Mohs micrographic surgery. The depth and extent of tumor infiltration were evaluated before the surgery. One day before the surgery, 20% aminolevulinic acid hydrochloride was topically applied to determine and label surgical margins under a Wood′s lamp. After fluorescence-based localization, peritumoral skin tissues were resected and underwent frozen-section examination according to the protocol for Mohs micrographic surgery. Meanwhile, the tumor was resected. After surgery, patients were followed up every 3 - 6 months to detect local recurrence and metastasis. Results Of the 28 patients, 25 were male and 3 were female. Six patients each underwent 3 sessions of frozen-section examination, and 12 patients each received 2 sessions, with an average of 1.86 sessions for each patient. During the follow-up for 5 - 72 months, local recurrence occurred in 3 cases, and 1 patient died of tumor metastasis and uremia after 2 years of follow-up. Conclusion Peripheral Mohs micrographic surgery is a time-saving and effective treatment for EMPD.

Objective To investigate the minimal persistent pigment dose (MPPD)of normal skin to UVA in Guangzhou city,and to observe its relationship to sex,age,skin type,seasons,ITA,and the years lived in Guangzhou.Methods 316 healthy subjects were exposed to Solar 601-300,and the MPPD was measured and observed by two professional technicians after 2-3 hours.Results The average MPPD value of all subjects was (9.61±2.57) J/cm2.In male and female,the average MPPD values were (11.09 ± 2.82) J/cm2 and (9.01 ± 2.20) J/cm2 respectively,and male was significantly higher than female (P＜0.01).There was significant difference of the MPPD value in different seasons (P＜0.01).The MPPD value in winter (10.66± 2.71) J/cm2) was significantly higher than spring (9.37±2.39) J/cm2,summer (9.53±2.66) J/cm2 and autumn (8.98±2.25) J/cm2.There was significant difference of the MPPD value between different ITA groups (P＜0.01).ITA grade-3 (10.72± 2.84) J/cm2 was significantly higher than grade-1 (8.50±1.45) J/cm2 and grade-2 (9.12±2.31) J/cm2 (P＜0.01),but there was no significant difference from grade-4 (11.87±2.73) J/cm2 (P =0.93).The MPPD value in subjects lived in Guangzhou over 10 years (8.97± 1.88) J/cm2 was significantly lower than those whose residence was less than 1 year and from 1 to 5 years,respectively (P＜0.01),but there was no significant difference from those lived from 5 to 10 years (P =0.47).Conclusions The MPPD value of the subjects in Guangzhou is related to gender,seasons,ITA grade and the years lived in Guangzhou city,while there is no correlation with age and skin types.

Objective To investigate the role of cathepsin G in photoaged fibroblasts. Methods Human fibroblasts were cultured and induced to premature senescence using UVA + MOP methods. Senescence-associated-β-galactosidase (SA-β-gal) stain was used to evaluate the positive rate of aged cells. The mRNA and protein expression of cathepsin G in photoaged fibroblasts were detected by real-time RT-PCR and Western blot techniques. Results Over 98 % induced cells presented a positive SA-β-gal straining. The expression of cathepsin G, detected by Western blot, was increased to (1. 70±0. 028) times of the control. And RT-PCR revealed that the synthesis of cathepsin G mRNA was also up-regulated to 1. 42±0. 09. Conclusion The results of our study demonstrates a significant correlation between photoaged fibroblasts and cathepsin G. The up-regulation of cathepsin G may play an important role in the damages of extracellular matrix and activation of MMPS in photoaged human skin.

Objective To study the effect and mechanism of bone marrow mesenchymal stem cells (BMSCs) on autologous peripheral blood lymphocyte proliferation in vitro in patients with hepatitis B virus related decompensated cirrhosis.Methods MSCs were isolated and expanded from human bone marrow blood of nine patients with decompensated cirrhosis,four groups were designed for experiment:①BMSCs + lymphocytes + PHA (contact co-culture) ; ② BMSCs + lymphocytes + PHA (non-contact co-culture) ; ③lymphocytes + PHA (positive control) ; ④lymphocytes alone (negative control).Lymphocytes proliferation rate were detected by flow cytometry.The mRNA expression levels of interleukin-10 (IL-10)and transforming growth factor-β1 (TGF-β1)in BMSCs were detected by using reverse transcription-polymerase chain reaction (RT-PCR).Results Cell proliferation of contact and non-contact co-culture groups significantly declined when compared with that of positive control group (all P ＜ 0.01),The relative expression levels of IL-10 mRNA and TGF-βlmRNA in BMSCs of contact and non-contact co-culture group raised up significantly after culture (all P ＜ 0.01).Besides,there was no significant difference on lymphocyte proliferation rate or expression levels of IL-10mRNA and TGF-β1 mRNA between contact co-culture group and non-contact coculture group (all P ＞ 0.05).Conclusion BMSCs from cirrhotic patients can inhibit proliferation of autologous peripheral blood lymphocytes,the mechanism may be associated with the secretion of inhibitory cytokine IL-10 and TGF-β1 in BMSCs.

Objective To study the effect of bone marrow mesenchymal stem cells (BMSCs) on autologous peripheral blood lymphocyte proliferation and regulatory T cells (Tregs)subsets in vitro in patients with hepatitis B virus related decompensated cirrhosis.Methods MSCs were isolated and expanded from human bone marrow blood of eight patients with decompensated cirrhosis.The purity of MSCs was identified by flow cytometry.lymphocytes were isolated from the peripheral blood of patients and stained with carboxy fluoresce indiacetate succinimidyl ester (CFDA SE).The BMSCs and peripheral blood lymphocytes from patients were added into wells containing autologous serum in the presence of phytohemagglutinin (PHA).lymphocytes proliferation rate and CD4 + CD25 + CD127-Tregs frequency were detected by flow cytometry.Results Flowcytometric analysis showed that lymphocyte proliferation in contact co-culture group and noncontact co-culture group were much lower than positive control (all P ＜ 0.01),and significantly higher than negative control (all P ＜ 0.01),while CD4+ CD25+ CD127-Tregs in contact co-culture group and noncontact co-culture group were much higher than positive control and negative control (all P ＜ 0.01).Besides,there was no significant difference on lymphocyte proliferation rate or Tregs frequency between contact co-culture group and non-contact co-culture group(all P ＞ 0.05).Conclusion BMSCs can inhibit proliferation of autologous peripheral blood lymphocytes and increase expression of CD4 + CD25 + CD127-Tregsin patients with decompensated cirrhosis.