Effect of Draconis Sanguis-containing serum on NGF, BDNF, CNTF, LNGFR, TrkA, GDNF, GAP-43 and NF-H expressions in Schwann cells / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of Draconis Sanguis-containing serum on the expressions of NGF, BDNF, CNTF, LNG-FR, TrkA, GDNF, GAP-43 and NF-H in Schwann cells, and investigate the possible mechanism of Draconis Sanguis to promote peripheral nerve regeneration.</p><p><b>METHOD</b>SD rats were randomly divided into 2 groups: the Draconis Sanguis group (orally administered with Draconis Sanguis-containing balm solution) and the blank group (equivoluminal balm) to prepare Draconis Sanguis-containing serum and blank control serum. Schwann cells were extracted from double sciatic nerves of three-day-old SD rats, divided into 2 groups: the Draconis Sanguis group and the blank control group, and respectively cultured with 10% Draconis Sanguis-containing serum or blank control serum. The mRNA expressions of NGF, BDNF, CNTF and other genes in Schwann cells were measured by RT-PCR analysis 48 hours later.</p><p><b>RESULT</b>Most of the Schwann cells were bipolar spindle and arranged shoulder to shoulder or end to end under the microscope and identified to be positive with the immunocytochemical method. To compare with the blank group, mRNA expressions of NGF, LNGFR, GDNF and GAP-43 significantly increased (P < 0.01). Whereas that of BDNF decreased significantly (P < 0.05), and so did that of TrkA, CNTF (P < 0.01), with no remarkable difference in NF-H-mRNA.</p><p><b>CONCLUSION</b>Traditional Chinese medicine Draconis Sanguis may show effect in nerve regeneration by up-regulating mRNA expressions of NGF, LNGFR, GDNF and GAP-43 and down-regulating mRNA expressions of TrkA, BDNF and CNTF.</p>

Fingerprint of Plantaginis Semen and determination of its two index contents / 中草药

Objective: To establish the fingerprint of Plantaginis Semen and to determine the two index contents of geniposidic acid and verbascoside. Methods: The chromatographic separation was performed on a C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase was a mixture of acetonitrile-0.5% acetic acid aqueous solution in gradient elution. The flow rate was 1.0 mL/min, and the detection wave length was 254 nm. Similarity evaluation system was used in data analysis. Results: The HPLC fingerprint of Plantaginis Semen showed 13 characteristic peaks with high similarity, and contents of geniposidic acid and verbascoside were 0.794 3% and 0.664 2%, respectively. Conclusion: The method that combining the determination of index contents with fingerprint analysis is timesaving and could be used for the quality control of Plantaginis Semen. It could provide a reference for the further research on the quality control of formular granule with Plantaginis Semen extracts.

Effect of storax on concentration of sulpiride in brain and blood by MD-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of traditional Chinese medicine storax on the concentration of combined western medicine sulpiride in brain and blood, discuss the effect of storax in inducing resuscitation and increasing the permeability of the gastrointestinal barrier (GB) and the blood-brain-barrier (BBB), and explore the interaction between storax and sulpiride.</p><p><b>METHOD</b>Rats were orally administered with the drugs for one week, probes were implanted in their brains and necks by surgery. After balance for 60 min, brain microdialysis and blood microdialysis were adopted for collect dislysates from blood in right atrum and cerebral hippocampus at time periods of 30, 60, 90, 120, 150, 180 min. The concentration of sulpirde in the samples was detected by RP-HPLC. Statistical approaches were adopted to compare the contents of sulpirde in brain and blood of the two groups.</p><p><b>RESULT</b>The sulpiride combined with storax group showed a significant higher concentration of sulpiride than the pure sulpiride group. The pure sulpiride group showed a concentration ratio between sulpiride in brain and blood of 1:0.2; while the sulpiride combined with storax group increased the concentration ratio between sulpiride in brain and blood to 1:0.3. Compared with the pure sulpiride group, the sulpiride combined with storax group showed an increase of concentration by 39% in brain and 69% in blood.</p><p><b>CONCLUSION</b>Storax can notably increase the concentration of sulpiride in rat brain and blood, indicating that it can increase the permeation of sulpiride through gastrointestinal barrier and BBB. This study reveals the mechanism of storax in inducing resuscitation by promoting the permeation through gastrointestinal barrier and BBB.</p>

Effects of human ERMAP-siRNA on erythroid differentiation of K562 cells induced by Ara-C / 中国实验血液学杂志

In order to investigate the potential role of human ERMAP gene in erythropoiesis, the ERMAP-dsDNA was designed, ERMAP-shRNA expressing plasmids was constructed, and ERMAP-shRNA/K562 cell was established. Cell morphology, biphenylamine staining, expression of cell surface antigens as well as quantitative level of human ERMAP gene were observed during K562 cells differentiating toward erythroid lineage induced by Ara-C. The results showed that at 72 hours after Ara-C treatment, ERMAP-shRNA/K562 cell size became large with increasing cytoplasm content. The percentage of biphenylamine positive cells increased from 1.17% to 2.04% (p < 0.05), but still lower than that in group K562 + Ara-C. The percentage of CD36(-)/CD235a(+) increased from 8.83% to 11.28%, CD36(+)/CD235a(+) increased from 1.23% to 2.64%, and CD36(+)/CD235a(-) increased from 0.59% to 1.47% respectively, which were all lower than that in group K562 + Ara-C at either time point. At the same time, the level of ERMAP expression increased slowly from 2.52 x 10(-3) to 4.53 x 10(-3), which was also significantly lower than that of group K562 + Ara-C. It is concluded that the ERMAP-shRNA inhibits the Ara-C-induced erythroid differentiation of K562 cells, which further suggests that there is relationship between hERMAP and erythroid differentiation and development.

Expression of human ermap gene in umbilical cord blood mononuclear cells during differentiation and development towards erythroid lineage / 中国实验血液学杂志

The aim of study was to explore the potential of human erythroid membrane associated protein (ERMAP) gene in erythroid cell differentiation and development, mononuclear cells (MNCs) were isolated from umbilical cord blood and induced to erythroid cell differentiation by SCF, IL-3 and EPO. The cell morphology was observed by using optical microscopy, the positive rate of cells was counted by biphenylamine staining and the ratios of CD36+/CD235a-, CD36+/CD235a+, CD36-/CD235a+ cells were detected by flow cytometry, the change of human ermap gene expression level was analyzed by using fluorescent quantitative PCR (FQ-PCR). The results showed that the ermap gene expression level increased while MNCs were induced to erythroid lineage after treatment with SCF, IL-3 and EPO. It is concluded that the human ermap gene plays an important role in differentiation and development of erythroid cells.

Research for dependablity of administration of platycodi radix in Tianwang Buxinwan decoction with change of brain inhibitive neurotransmitte in rats by microdialysis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of Tianwang Buxinwan decoction on the contents of amino acids neurotransmitters in corpus striatum of rats to implicate the mechanism of Tianwang Buxinwan promoting and Improving sleeping.</p><p><b>METHOD</b>Contents of two amino acids neurotransmitters in corpus striatum of rats were prepared by microdialysis technology and were determined by HPLC which involved pre-column derivation with orthophthaladehyde, recersed-phase gradient elution and fluorescence detection.</p><p><b>RESULT</b>In the experimental separation condition, Tianwang Buxinwan seemed do not influence three kinds of contents of amino acids neurotransmiters (glutamic acid, glycin, aspartic acid), but TBW seemed increase the content of gamma-GABA in corpus striatum of rats.</p><p><b>CONCLUSION</b>The effects of Tianwang Buxinwan to relieve uneasiness may relate with the inhibitory amino acids gamma-GABA. Tianwang Buxinwan may promote increasing the content of gamma-GABA. This discovery may be helpful for the deep study of related mechanism of Tianwang Buxinwan.</p>

Expression of human ERMAP gene in fetal tissues / 中国实验血液学杂志

Human ERMAP (hERMAP) is a novel gene coding for erythroid membrane-associated protein, which may play a role in erythropoiesis. To explore the role of hERMAP in fetal hematopoiesis, 29 fetuses of inevitable abortion with the fetal ages of 9 - 36 weeks were collected, and the total RNAs were isolated from the liver, spleen, kidney, heart, lung, thymus, brain, bone marrow and skeletal muscle, the RNAs from which at 25(+5) week fetal age were used to detect hERMAP expressions in different fetal tissues with Northern blot, and the hERMAP in various fetal tissues were quantified by FQ-PCR. As a result, Northern blot showed that hERMAP was expressed in liver and bone marrow at 25(+5) fetal age, but not in the other organ tissues. FQ-PCR results indicated that the hERMAP had been still expressed in 9 - 36th week fetal liver, increased starting from 12th-week, reaching a peak between 18 - 20th week and declining slowly starting from 21st-week. In the fetal bone marrow, expression of the hERMAP began from 15th week, reached a highest level between 27 - 32nd week and fell rapidly from 33rd week, the expression level of which was lower than that in liver. The low level expression of this gene was observed in some specimens of kidney, heart, lung, thymus, brain and skeletal muscle. It is concluded that the expression of hERMAP in fetal liver approximately consistent with haematopoiesis of fetal liver, and the expression of this gene in bone marrow aged 15 - 32nd fetal weeks coincides with haematopoiesis of fetal bone marrow. It suggests that the function of the hERMAP is possibly related with the migration of erythroid cells to liver and bone marrow during the fetal development process.

RNA isolation from human embryonic tissues / 中国实验血液学杂志

To investigate the method of RNA isolation from human embryonic tissues and the factors influencing the quality of RNA, the RNA from human embryonic tissues obtained with drug-induced labor or non-drug induced labor were isolated by using grind with liquid nitrogen or homogenizer without liquid nitrogen. The results showed that the positive rates of RNA integrity in grind with liquid nitrogen group and in homogenizer without liquid nitrogen group were 68.42% and 29.79% respectively, and there was significant difference between these two groups; however, there was no statistic difference in positive rate of RNA integrity, OD(260)/OD(280) ratio and beta-actin gene expression level between the drug-induced labor group and non-drug induced labor group. It is concluded that pulverize of tissue in liquid nitrogen remains the integrity of RNA isolated and may be applied for RNA isolation from human embryonic tissues. The quality of RNA is not affected by different methods of induction of maternal labor.