Effect of liquorice decoction on rat intestinal P-glycoprotein / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of liquorice in functional modulation of intestinal P-glycoprotein (P-gp) in rats.</p><p><b>METHODS</b>An in vitro diffusion chamber system (Ussing chamber) was used to examine the direct effect of liquorice decoction on rhodamine 123 (a subtrate of P-gp) transport and evaluate the permeability of rhodamine 123 or fluorescein sodium through rat jejunum membranes after oral administration of liquorice decoction.</p><p><b>RESULTS</b>Direct application of liquorice decoction did not obviously affect rhodamine 123 transport across the intestinal mucosa. Oral administration of liquorice decoction (10 g/kg, twice daily for a week) significantly increased the absorption of rhodamine 123 and also enhanced rhodamine 123 secretion across the jejunum mucosa. Liquorice had no obvious effect on the transport of CF across the jejunum mucosa.</p><p><b>CONCLUSION</b>Liquorice may slightly inhibit P-gp function in the intestinal mucosa to increase the intestinal absorption of rhodamine 123.</p>

Study on Transplantation of Human Umbilical Cord Mesenchymal Stem Cells into Infertile Mouse Testis / 实用儿科临床杂志

Objective To observe clone ability of human umbilical cord mesenchymal stem cells (MSCs) into infertile mouse seminife-rous tubules and the effects of MSCs on reproductive function.Methods Busulfan was used to destroy endogenous spermatogenesis of the recipient mice.To isolate,culture and purify MSCs with adherent method before marked with Brdu and Hoechst 33258 respectively,and then transplanted into the seminiferous tubules by microinjection.The survival of MSCs in recipient testes were evaluated by immunohistochemistry stained for Brdu and Fluorescent microscopy for Hoechst 33258 observation at different times.The diameter of seminiferous tubules was detected with HMIAS-2000 high-definition colored analyzing system for medical pictures.SPSS 13.0 software was used to analyze the data.Results The dosage of Busulfan resulted in 15% death in the mice,the testis of survived mice showed only basilar membrane in seminiferous tubules after 4 weeks.A lot of purified MSCs were obtained at the third generation and transplantation them into mouse seminiferous tubules survive for at least 4 months and appear to migration.The average diameter in experimental groups were higher than those in controls not only on 26 days but also on 120 days(P

Human Umbilical Cord Blood Mesenchymal Stem Cells Colonizing in Hypoxic-Ischemic Encephalopathy Rats Brain / 实用儿科临床杂志

Objective To explore human umbilical cord blood mesenchymal stem cells(MSCs)colonization in hypoxic-ischemic encephalopathy(HIE)rats brain.Methods Models of 7-day-old newborn rats with HIE brain injury were established.Meanwhile,on the same day,MSCs were transplanted with Hoechst 33258 for 24 hours into rats models marked by flurescent nucleotide dye injected through caudal vein or with stereotactic instrument.After 15-30 days,then MSCs were detected with fluorescene microscope.Results With the improved rice methods,HIE animal model was successfully attained.Majority of MSCs were distributed in the cortex,hippocampus of the lesioned hemisphere,especially in the forehead.And abtained a good fusion with HIE rats brain tissue.Conclusion Human umbilical cord blood MSCs can be cultured,when transplanted into the HIE rats model,they can move into intracranial,and integer with rats brain tissue.

Expression of hepatitis B virus genes in early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To detect the expression of hepatitis B virus (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique.</p><p><b>METHODS</b>Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome.</p><p><b>RESULTS</b>Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences.</p><p><b>CONCLUSION</b>Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.</p>

Biological characteristics of human umbilical cord-derived mesenchymal stem cells and their differentiation into neurocyte-like cells / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the isolation and expansion of mesenchymal stem cells (MSCs) from human umbilical cord Wharton's jelly and their biological identities, and explore the possibility of inducing human umbilical cord-derived MSCs to differentiate into neurocyte-like cells.</p><p><b>METHODS</b>The growth and proliferative abilities of human umbilical cord-derived MSCs were observed, and their immunophenotypes were determined by flow cytometry. Salvia miltiorrhiza and beta-sulfhydryl alcohol were adopted to induce the cells to differentiate. The differentiated and undifferentiated cells were identified with immunocytochemistry. The pleiotrophin and nestin genes were measured by RT-PCR.</p><p><b>RESULTS</b>A population of human umbilical cord-derived MSCs were isolated from human umbilical Wharton's jelly; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. The human umbilical cord-derived MSCs were positive for CD(29), CD(44), CD(59), CD(105), but negative or weakly expressed the markers of hematopoietic cells such as CD(14), CD(33), CD(34), CD(28), CD(45) and CD(117). The important GVHD correlation markers were negative or weakly expressed, including CD(80) (B7-1), CD(86) (B7-2), CD(40) and CD(40L). Salvia miltiorrhiza beta-sulfhydryl alcohol could induce the MSCs to express nestin, a marker of neuronal precursor stem cells at early stage of differentiation. Later, they exhibited neural phenotypes, expressing beta-tubulin III and neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that the MSCs could express pleiotrophin either before or after the induction of salvia miltiorrhiza, furthermore, after the induction the expression was markedly enhanced and the nestin gene was also expressed.</p><p><b>CONCLUSION</b>The human MSCs could be isolated from human umbilical cord Wharton's jelly, and it was easy to propagate these MSCs. The negative GVHD correlated markers might result from the fact that MSCs had no HLA barrier, which may suggest potential clinical significance. The MSCs are capable of differentiating into neurocyte-like cells and they may represent an alternative stem cell source for CNS cells transplantation.</p>

Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells differentiation into nerve-like cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The two most basic properties of mesenchymal stem cells (MSCs) are the capacities to self-renew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton's Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton's Jelly-derived MSCs to differentiate into nerve-like cells.</p><p><b>METHODS</b>MSCs were cultured from the Wharton's Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and beta-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate. The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy (EM), and laser scanning confocal microscopy (LSCM). The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>MSCs in the Wharton's Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease (GVHD)-related cells. Treatment with Salvia miltiorrhiza caused Wharton's Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia miltiorrhiza and beta-mercaptoethanol also induced MSCs to express nestin, beta-tubulinIII, neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed.</p><p><b>CONCLUSIONS</b>MSCs could be isolated from human umbilical cord Wharton's Jelly. They were capable of differentiating into nerve-like cells using Salvia miltiorrhiza or beta-mercaptoethanol. The induced MSCs not only underwent morphologic changes, but also expressed the neuron-related genes and neuronal cell markers. They may represent an alternative source of stem cells for central nervous system cell transplantation.</p>

Effect of dynein inhibitor on mouse oocyte in vitro maturation and its cyclin B1 mRNA level / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the effect of dynein inhibitor on mouse oocyte in vitro maturation and its cyclin B1 transcription level.</p><p><b>METHODS</b>Immature mouse oocytes were cultured in vitro with a known dynein ATPase activity inhibitor-sodium orthovanadate (SOV) to detect the changes of maturation rate, and semi-quantitative RT-PCR and single cell RT-PCR were performed to detect the changes of cyclin B1 mRNA level.</p><p><b>RESULTS</b>In dose-dependent experiments, the maturation rates of oocytes were significantly different between 5 micromol/L SOV and control groups (P < 0.05), and decreased with SOV increasing doses. However, the elevation of cyclin B1 mRNA level of immatured oocytes cultured for 12 h depended on SOV concentrations ranging from 50 to 500 micromol/L. In incontinuity exposed SOV experiments, the maturation rates of oocytes markedly reduced after the first incubation with 400 micromol/L SOV at least for 1 h and were first cultured in SOV-free medium for 4 h or 8 h before exposure to SOV (P < 0.05). In time-course experiment, the opposite changes of cyclin B1 mRNA level in oocytes between SOV and control groups were observed.</p><p><b>CONCLUSION</b>Dynein inhibitor might delay oocytes meiosis process, and cause ectopic expression of cyclin B1 in oocytes. Most Oocytes incubated with SOV blocked at germinal vesicles (GV) stage or M I to anaphase transition due to dynein dysfunction and ectopic transcription level of cyclin B1.</p>

Biological Characteristics of Mesenchymal Stem Cells Cultured in Conditions for Spermatogonia in Vitro / 实用儿科临床杂志

Objective To investigate the possibility of inducing mesenchymal stem cells(MSCs)from human umbilical cord Wharton's Jelly to differentiate into spermatogonia.Methods To isolate,culture and purify MSCs with adherent method,the growth and proliferation of human umbilical cord-derived MSCs were observed,and their immunophenotypes were determined by flow cytometry;MSCs of the third generation were divided into 2 groups to be induced and cultured,MSCs of the control group were cultured in basal medium,while those of the experimental group with conditional medium.The morphologic and ultrastructure changes of control group and experimental group cells were compared with phase contrast microscopy,electron microscopy(EM)and transmission electron microscope(TEM)respectively ;the spermatogonial cells differentiated were then evaluated by immunohistochemistry stained for CD117and CD49f ;the method of Western-blot was used to test if the cells induced could express CD49f.Results A population of MSCs were isolated from human umbilical Wharton's Jelly;they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling;After induction,the shape of MSCs changed greatly from the fibroblast to the round,even familiar to the tadpole;expressed the known molecular markers of spermatogonial cells,such as CD49f,CD117.Conclusion The induced MSCs not only undergo spfermatogonial-cell like morphologic changes,ultramicrostructure mature with increasing cell organs,but also express the spermatogonial cell markers,which show that human umbilical cord derived MSCs are capable of differentiating into spermatogonial cell.