Anatomical observation and transcriptome analysis of buds reveal the association between the AP2 gene family and reproductive induction in hybrid larch (Larix kaempferi × Larix olgensis).

Hybrid larch is an excellent afforestation species in northern China. The instability of seed yield is an urgent problem to be solved. The biological characteristics related to seed setting in larch are different from those in angiosperms and other gymnosperms. Studying the developmental mechanism of the larch sporophyll can deepen our understanding of conifer reproductive development and help to ensure an adequate supply of seeds in the seed orchard. The results showed that the formation of microstrobilus primordia in hybrid larch could be observed in anatomical sections collected in the middle of July. The contents of endogenous gibberellin 3 (GA3) and abscisic acid (ABA) were higher and the contents of GA4, GA7, jasmonic acid and salicylic acid were lower in multiseeded larch. Transcriptome analysis showed that transcription factors were significantly enriched in the AP2 family. There were 23 differentially expressed genes in the buds of the multiseeded and less-seeded types, and the expression of most of these genes was higher in the buds than in the needles. We conclude that mid-July is the early stage of reproductive organ development in hybrid larch and is suitable for the study of reproductive development. GA3 and ABA may be helpful for improving seed setting in larch, and 23 AP2/EREBP family genes are involved in the regulation of reproductive development in larch.

Enhanced production of ethyl acetate using co-culture of Wickerhamomyces anomalus and Saccharomyces cerevisiae.

The style and quality of Baijiu is greatly influenced by ethyl acetate. Therefore, improving and controlling the ethyl acetate levels in Baijiu is important. This study investigated ethyl acetate production using a co-culture of Saccharomyces cerevisiae Y3401 and Wickerhamomyces anomalus Y3604. More ethyl acetate was produced in mixed fermentations using both yeasts than in single fermentations. The highest ethyl acetate yield was 6.41 g/L using a Y3401/Y3604 ratio of 3:1. Synergistic fermentation using both yeasts not only improved ethyl acetate production, but also increased the contents of other flavor compounds, such as ß-phenethyl alcohol and phenethyl acetate. Therefore, the co-culture of S. cerevisiae and W. anomalus had a positive effect on ethyl acetate production and provides opportunities for altering the aroma and flavor perception of Baijiu.

Improving Ethyl Acetate Production in Baijiu Manufacture by Wickerhamomyces anomalus and Saccharomyces cerevisiae Mixed Culture Fermentations.

Ethyl acetate content has strong influence on the style and quality of Baijiu. Therefore, this study investigated the effect of Saccharomyces cerevisiae Y3401 on the production of ethyl acetate by Wickerhamomyces anomalus Y3604. Analysis of cell growth showed that Y3401 influences Y3604 by nutrient competition and inhibition by metabolites, while the effect of Y3604 on Y3401 was mainly competition for nutrients. Mixed fermentation with two yeasts was found to produce more ethyl acetate than a single fermentation. The highest yield of ethyl acetate was 2.99 g/L when the inoculation ratio of Y3401:Y3604 was 1:2. Synergistic fermentation of both yeasts improved ethyl acetate production and increased the content of other flavor compounds in liquid and simulated solid-state fermentation for Baijiu. Saccharomyces cerevisiae had a positive effect on ethyl acetate production in mixed culture and provides opportunities to alter the aroma and flavor perception of Baijiu.

[Experimental Research of Circular RNA hsa_circ_0000254 in Acute Myeloid Leukemia].

OBJECTIVE: To clone the circular RNA hsa_circ_0000254 and construct its lentiviral over-expression vector. METHODS: The sequence of hsa_circ_0000254 (a total of 524 bp long) was synthesized and cloned by using pGH vector. The vector was cut by EcoR I and BamH I, and artificial hsa_circ_0000254 was obtained, then inserted in pLCDH-ciR to construct the recombinant expression vector pLCDH-circ254(C254), which was confirmed by DNA sequencing. The lentiviral expression vectors pLCDH-circ254(C254) and NC(pLCDH-ciR) were cotransfected into 293T cells by lipofectamineTM 2000(lipo2000). After transfection for 40 hours, the cells were collected and verified by PCR and sequencing. RESULTS: Restriction analysis and DNA sequencing demonstrated that the lentiviral vector pLCDH-circ254(C254) was constructed successfully, the expression efficiency increased 10000 times after transfection of cells. CONCLUSION: The successful construction of the lentiviral expression vector pLCDH-circ254(C254) results in the production of high-titer lentivirus, so as to facilitate further study of the molecular functions of hsa_circ_0000254.

Mindset switching increases the use of 'want-based' over 'should-based' behaviors.

This study examines the consequences of mindset switching on behavioral choices in want/should conflicts. Building on the insights of the ego depletion literature, we propose that mindset switching depletes individuals' self-control resources and therefore prompts the choice of want behavior, which provides immediate pleasure, over should behavior, which provides long-term utility. Four laboratory experiments with university students that stimulated individuals to switch mindsets were conducted to test our hypotheses. Experiment 1 demonstrated that switching between individualist and collectivist mindsets increased the subjects' tendency to prefer popular magazines over scientific journals. Experiment 2 replicated the results by testing the relationship between an abstract/concrete mindset-switching task and want/should online behavioral choices. The mediating effect of ego depletion was also supported. Experiment 3 retested the main effect of language-switching on reading choices, and the mediating effect of ego-depletion. Experiment 3 also tested the moderating effect of the Need for Cognition, and eliminated the alternative explanation of cognitive fatigue. In Experiment 4, actual food choices were used as the direct measure of want/should behaviors to test the robustness of our findings. The results consistently supported our hypotheses that mindset switching has significant effects on behavioral choices in terms of overindulgence, such as increasing want behavior and thus foregoing should behavior.

Effects of sodium copper chlorophyllin on mesenchymal stem cell function in aplastic anemia mice.

OBJECTIVE: To investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia. METHODS: A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×10(6) lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia. RESULTS: Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3(+)) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor ß1 was up-regulated in SCC-treated groups. CONCLUSION: The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.

Effects of Panax notoginseng saponins on proliferation and differentiation in NIH3T3 cells.

OBJECTIVE: To investigate the effects of Panax notoginseng saponins (PNS) on the proliferation and differentiation in NIH3T3 cells. METHODS: NIH3T3 cells were treated by various concentrations of PNS 0, 0.05, 0.10, 0.20, and 0.40 g/L. The vitality and proliferation potential of cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, the alkaline phosphatase (ALP) activity was measured by p-nitrophenyl phosphate (pNPP) assay, and the mineralization formation ability was tested for the cellular differentiation toward osteoblast, as well as the expression level of phosphorylated extracellular signal-regulated kinase1/2(P-ERK1/2), extracellular signal-regulated kinase1/2 (ERK1/2) protein kinase was analyzed by Western blot with total cell lysate of NIH3T3 cells treated by PNS. RESULTS: Both MTT and pNPP assay showed that optical density (OD) values were increased in response to PNS treatment at a dose-dependent pattern. The mineralization formation ability was enhanced in PNS-treated NIH3T3 cells compared with untreated cells. Meanwhile, the expression level of P-ERK1/2 protein kinase was up-regulated in PNS-treated NIH3T3 cells, while, the expression level of ERK1/2 protein kinase revealed no obvious difference with or without PNS treated cells. CONCLUSION: PNS could pay a role to promote the proliferation and differentiation in NIH3T3 cells by means of up-regulation of P-ERK1/2 protein kinase.

[Retrospective study on forty cases of medical legal dispute].

OBJECTIVE: To analyze the main causes of medical dispute and the main types of medical malpractice. The related problems were discussed in forensic expertise. METHODS: Forty cases of medical dispute from 2006 to 2008 in our institute were analyzed retrospectively. RESULTS: In 40 cases of medical dispute, city-level hospitals, county-level hospitals, town-level hospitals and private clinics were 11 (27.5%), 24 (60.0%), 2 (5.0%) and 3 (7.5%) cases respectively. The internal medicine departments, surgical departments, gynaecological and obstetric departments, pediatric departments and outpatient center were 16 (40.0%),10 (25.0%), 9 (22.5%), 2 (5.0%) and 3 (7.5%) cases, respectively. The amount of cases from city-level hospitals, county-level hospitals diagnosed by the medical experts as the medical malpractice showed less than that from town-level hospitals and private clinics. CONCLUSION: The amount of cases of medical dispute from city-level and county-level hospitals were more than that of town-level hospitals. But the amount of cases diagnosed by the medical experts as medical malpractice from city-level and county-level hospitals were less than that of town-level hospital and private clinics.

[Characteristics of death cases caused by road traffic accident in Shenzhen].

OBJECTIVE: By summarize the characteristics of death cases caused by road traffic accident, to provide information and data for prevention of traffic accident. METHODS: To retrospectively analyze 4148 death cases caused by road traffic accident in Shenzhen. The characteristics studied include the age and sex of the dead, the cause of death, time and place of the accidents and vehicle types, etc. RESULTS: The death was mainly male and the proportion of male to female is 2.45:1; the accidents mainly occured in 6:00-8:00 and 18:00-2:00; 72% of the accidents took place on the main suburban roads. The proportions for each traffic modes of the death were: 44% of walking, 19% of bike, 15% of motorbike. Most traffic accidents were induced by truck, 83.2% of the death causations were severe head injury and 13.3% were complex injuries. CONCLUSION: The death cases of road-traffic accident in Shenzhen has obvious characteristic and maybe is preventible.

[Inhibition effect of hedyotis diffusa wild injection on HL-60 cells and its mechanism].

This study was aimed to explore the inhibition effect and mechanism of hedyotis diffusa wild injection (HDI) on leukemia cell line (HL-60) in vitro. The leukemia cell line HL-60 was used as target cells. The inhibitory effects of HDI on proliferation of HL-60 cells were observed by MTT assay. The positive rate of cell apoptosis and the surface marker of granulocytic differentiation (CD33 and CD15) were measured by flow cytometry. The expressions of anti-apoptosis related gene (survivin and bcl-2) were detected by RT-PCR. The results showed that the growth of HL-60 cells was inhibited by higher concentration of HDI (3.12 - 12.5 ml/L) and inhibited obviously in dose-dependent manner (p < 0.05 or p < 0.01), but not suppressed by low concentration of HDI (1.56 ml/L) in liquid culture system (p > 0.05). The FCM and DNA Ladder results showed that the phenomenon of typical apoptosis did not detected after HL-60 cells were treated with the different concentrations of HDI for 24, 48 and 72 hours respectively. After HL-60 cells were treated with HDI (1.56, 3.12, 6.25 and 12.5 ml/L) for one week, the expression level of CD15 surface marker was all enhanced obviously. When treated with HDI (6.25 ml/L) for 3 weeks, the expression levels of survivin and bcl-2 gene were also decreased obviously by 60% and 44% respectively. It is concluded that HDI can inhibit HL-60 cells in the presence of its higher concentrations. The mechanisms of HDI may induce HL-60 cells differentiation, and suppress the expression of anti-apoptosis related gene (survivin or bcl-2) to inhibit the growth of HL-60 cells.

[Apoptosis of HL-60 cells induced by aescinate].

The aim of this study was to investigate the effects of aescinate on inhibition and apoptosis of HL-60 cell line from promyelocytic leukemia. HL-60 cells at logarithm phase were treated with aescinate. Cell survival rate and cell morphology were observed, and the cell apoptosis was analyzed by Annexin V/PI-FITC double labeling and DNA electrophoresis. The results showed that HL-60 cells could be inhibited in the presence of 15-120 mg/L of aescinate for 48 hours, survival rates were (92.2+/-0.69)%-(8.2+/-0.96)%, which were significantly lower than that of non-aescinate control (99.4+/-0.31)% (all p<0.01). The apoptosis of cells could be induced by aescinate treatment at dosage of 15-60 mg/L for 24 hours, the Annexin V positive cells accounted for (12.7+/-0.58)%-(65.4+/-1.30)% which were significantly higher than that of non-aescinate control (0.57+/-0.03)% (all p<0.01). The typical DNA ladder of HL-60 cells treated with aescinate was shown on the DNA electrophoresis pattern. It is concluded that aescinate can specifically induce apoptosis of leukemic HL-60 cells, which provides an experimental evidence for treatment of leukemia with aescinate as a supplementary agent to chemotherapy.

Development and evaluation of real-time loop-mediated isothermal amplification for hepatitis B virus DNA quantification: a new tool for HBV management.

BACKGROUND: Accurate quantitation of hepatitis B viral load is a critical aspect in screening and monitoring HBV infection. OBJECTIVES: We used loop-mediated isothermal amplification (LAMP) to develop a real-time fluorogenic (RtF-LAMP) protocol to quantitate HBV DNA. Quantitative analysis was obtained by measuring time-to-positive (TTP), a biomarker similar to cycle threshold (Ct) in real-time PCR. STUDY DESIGN: Sensitivity, specificity, reproducibility, and dynamic range for RtF-LAMP were evaluated using molecular and biological standards. Four hundred and two patient samples were then used to compare the performance of RtF-LAMP to a commercial real-time PCR assay (DaAn Gene Co, China). RESULTS: The lower detection limit (LDL) of RtF-LAMP by Probit analysis at the 95% detection level was 210 copies/ml, and the dynamic range was 8 orders of magnitude. The conversion factor for results obtained with the RtF-LAMP assay was 1 IU/ml equals to 4.4 copies/ml. Coefficients of variation (CV) reflected low intra-assay and inter-assay variability (4.24-12.11%). In a large number of serum samples, there was excellent an correlation between RtF-LAMP and real-time PCR (R(2)=0.96). There was a good agreement between the two tests except at the detection cutoff of the real-time PCR assay. CONCLUSION: Our RtF-LAMP protocol appears to be precise, accurate and rapid. It could be a valuable tool for the detection of HBV in large clinical and epidemiological studies.

[HBVcccDNA of patients with hepatitis B in the serum, PBMC and liver tissue distribution].

OBJECTIVE: Hepatitis B virus covalently closed circular DNA in the serum of patients with hepatitis B, peripheral mononuclear cells (PBMC) and liver tissue distribution. METHODS: Serum HBV DNA > 10(5) copies/ml 50 cases of hepatitis B patients, serum HBV DNA < 10(5) copies/ml 30 cases of hepatitis B patients, patients with fatty liver (hepatitis B) 20 cases, using real-time quantitative polymerase chain reaction for detection of serum, and liver tissue in PBMC HBVcccDNA the existence. RESULTS: Of 50 serum HBV DNA > 10(5) copies/ml cccDNA the serum specimens were 28 cases, 56% detection rate, 29 cases were PBMC HBVcccDNA, 58% detection rate, liver tissue HBVcccDNA were 44 cases, 88% detection rate, serum, the PBMC were detected in liver tissue were significant differences in P < 0.005, compared serum PBMC were no significant differences in P > 0.005. 30 serum HBV DNA < 10(5) copies/ml the serum specimens, PBMC cccDNA detected two cases were 6.67% detection rate, liver tissue cccDNA were six cases 20% detection rate, serum, PBMC, were among the liver tissue was not significantly different P > 0.005. 20 in the serum of patients with fatty liver, liver tissue in PBMCwere not detected HBVcccDNA. CONCLUSION: HBVcccDNA mainly in the liver of patients with hepatitis B, hepatitis B patients and also cccDNA PBMC in the presence of liver tissue but a few of many.

Effects of notoginosides on proliferation and upregulation of GR nuclear transcription factor in hematopoietic cells.

AIM: To investigate the effects of panax notoginosides (PNS) on the proliferation of human hematopoietic stem/progenitor cells, and to explore the signaling pathway of the nuclear transcription factor of the glucocorticoid receptor (GR-NTF) initiated by PNS related with the proliferation. METHODS: The human CD34+ cells and bone marrow nuclear cells were exposed to PNS at a concentration of 0, 10, 25, 50, and 100 mg/L, respectively, in semi-solid culture system to observe colony forming unite of all lineages, granulocyte, erythrocyte, and megakaryocyte (CFUGEMM, CFU-GM, CFU-E, and CFU-MK). Three lineages of human hematopoietic cell lines, including granulocytic HL-60, erythrocytic K562, megakaryocytic CHRF- 288, and Meg-01 cells were incubated with PNS at 20 mg/L for 14 d. Meanwhile, dexamethasone (Dex) was used as a positive control. The nuclear protein of the cells was analyzed by Western blotting with monoclonal antibodies against the amino or carboxyl terminus of GR-NTF. Electrophoretic mobility shift assay performed by using the 32P-radiolabeled GR-NTF consensus oligonucleotide. RESULTS: PNS promoted the proliferation of CD34+ cells and significantly raised the colony numbers of CFU-GEMM by 34.7%+/-16.0% over the non-PNS control (P<0.01). PNS also enhanced the proliferation of CFU-GM, CFU-E, and CFU-MK by 39.3%+/- 5.7%, 33.3%+/-7.3%, and 26.2%+/-3.2%, respectively. GR-NTF protein levels of either the amino or carboxyl terminus in K562, CHRF-288, and Meg-01 treated by PNS increased by 2.4-2.8 fold and 1.3- 3.9 fold over the untreated cells. GR-NTF binding activity, initiated by either PNS or Dex, was apparently elevated to form the complex of GR-NTF with DNA as higher density bands in K562 and CHRF-288 cells, and some activity appeared as a band in HL-60 cells induced by PNS. CONCLUSION: PNS displayed the action of hematopoietic growth factor-like or synergistic efficacy to promote proliferation of human progenitor cells, may play a role in the upregulation of gene expression related to proliferation of hematopoietic cells through increasing the GR-NTF synthesis and its DNA binding activity.

[Expression of apoptosis-related proteins in the human bone marrow hematopoietic cells treated by Panax Notoginosides].

The study was aimed to investigate the action of Panax Notoginosides (PNS, extracted from notoginseng herb) on the expression of the apoptosis-related proteins (Daxx, Fas) and transcription factors (NFkB, c-Rel) in the hematopoietic cells and to explore the mechanisms of supporting cells to survive. The colony formation of CFU-GM and CFU-E in human bone marrow was assayed in the presence of various concentrations of PNS. The viability of cells was assayed by trypan blue and the changes of cell morphology were observed with microscope. The Annexin-V positive cells were detected by FCM. Three lineages of human myeloid HL-60, erythroid K562, megakaryoid CHRF-288 and Meg-01 cells were incubated in addition of PNS (10 mg/L) for 14 days. The nuclear or cytoplasm protein of cells was extracted and analyzed by Western blot with monoclonal antibodies against Daxx, Fas or NFkB, c-Rel. The results showed: (1) the proliferation on hematopoietic progenitor cells (CFU-GM and CFU-E) and four cell lines was promoted by PNS; (2) after the four cell lines were promoted by PNS and hungered through wiping off the sera, the viability of the four cell lines was high without significant morphological change and neither the detection of Annexin-V positive cells; (3) the expression of Daxx and Fas protein could be inhibited by PNS. Western Blot showed that Daxx in four cell lines treated by PNS were 33.3-61.5% lower than that in untreated controls. The Fas protein was also descended in three cell lines of K562, CHRF-288 and Meg-01 by 33.3-71.4% respectively, while Fas protein in HL-60 cells was no detectable difference after PNS treatment. (4) The transcription factors NFkB and c-Rel protein could be increased by PNS. The NFkB, c-Rel protein were also enhanced in three cell lines of K562, CHRF-288 and Meg-01 by (2.0-2.7) and (1.5-2.3)-fold respectively, while there were also no detectable difference in HL-60 cells after PNS treatment. It is concluded that PNS inhibites the expression of Daxx and Fas proteins, may decrease the apoptosis of the hematopoietic cells. The level of NFkB and c-Rel proteins can be enhanced by PNS, which not only stimulates the proliferation of cells, but also inhibits the activity of the waterfall of caspase and apoptosis of the hematopoietic cells. PNS may treat the disease with over-apoptosis of hematopoietic cells, as aplastic anemia.

High diversity of endophytic fungi from the pharmaceutical plant, Heterosmilax japonica Kunth revealed by cultivation-independent approach.

Heterosmilax japonica Kunth is well recognized for its diuretic effects in China. However, little is known about its endophytic fungi. In this study, microbial communities inhabiting the stems of H. japonica in spring and summer were investigated by light microscopy and cultivation-independent approaches, such as RFLP analysis and sequencing of rDNA ITS library. Molecular phylogenetic analysis showed that a broad spectrum of fungi, including Mycosphaerella, Phomopsis, Aureobasidium, Cladosporium, Glomerella, Botryosphaeria, Guignardia, is able to colonize the plants internally. Particularly, several rDNA sequences determined in this study like YJ4-61 are not specifically affiliated with any currently documented fungal sequences in the public database. Several sequence types, such as YJ4-9 and YJ4-70, are significantly similar to some uncultured environmental samples. Furthermore, our result also showed that the samples collected in spring harbored more abundant endophytic populations than that in summer, implying a seasonal fluctuation for the endophytes in H. japonica.

[Up-regulation of transcription factors NF-E2, c-jun and c-fos of AP-1 family induced by Panax Notoginosides in hematopoietic cells].

To observe the effects of Panax Notoginosides (PNS) on up-regulation of AP-1 family transcription factors NF-E2, c-jun and c-fos for exploring intracellular signal pathway of PNS in hematopoietic cells, four human hematopoietic cells lines including myeloid HL-60, erythroid K562, megakaryoid CHRF-288 and Meg-01 were incubated in the presence of PNS for 14 days. The nuclear protein of cells were extracted and analyzed by Western blot with antibodies against NF-E2, c-fos and c-jun. Electrophoretic mobility shift assay (EMSA) was performed by using (32)P labeled AP-1 consensus oligonucleotide which contains binding site for NF-E2, c-jun and c-fos. The results showed that the transcription factors NF-E2, c-jun and c-fos of AP-1 family could be induced by PNS. Western blot demonstrated that the nuclear protein of both NF-E2 and c-jun in four cell lines treated by PNS were increased by 1.5-2.5- and 2.0-3.0-fold over untreated cells respectively. The c-fos protein in three cell lines of K562, CHRF-288 and Meg-01 was also elevated by 2.0-3.0-fold respectively, while c-fos protein in HL-60 cells was no detectable difference after PNS treatment. EMSA results in four cell lines indicated that AP-1 binding activity initiated by PNS was apparently elevated to form higher density band of AP-1-DNA complex. In conclusion, the intracellular transcription regulation initiated by PNS was involved in transcription factors NF-E2, c-jun and c-fos of AP-1 family members, which could play an important role in the up-regulation of genes expression related to proliferation and differentiation of hematopoietic cells.

[Effect of panaxadiol saponin and panaxtrol saponin on proliferation of human bone marrow hemopoietic progenitor cells].

OBJECTIVE: To observe the effect of panaxadiol saponin (PDS) and panaxtrol saponin (PTS) on proliferation of human bone marrow hemopoietic progenitor cells (HPC). METHODS: PDS and PTS were separated and purified from ginsenosides, and the effects on HPC were studied using in vitro hemopoietic progenitor cell colony-forming technique, by observing the proliferation of human burst forming unit-erythroid progenitor (BFU-E), colony-forming unit-erythroid (CFU-E), colony-forming unit-granulocyte/macrophage (CFU-GM) and colony-forming unit-pluripotent hemopoietic progenitor (CFU-Mix) in mice after PDS and PTS stimulation. RESULTS: Different concentration of PDS (2.5-200 micrograms/ml) could stimulate the proliferation of HPC obviously, showing increase of CFU-E, BFU-E, CFU-GM and CFU-Mix by 54.9 +/- 6.3%, 48.8 +/- 5.1%, 27.6 +/- 4.2% and 48.9 +/- 3.9% respectively, which was higher than that of the control group. While stimulated by PTS of the same concentration, the CFU-E and BFU-E was lower than that of control significantly (P < 0.05); when the terminal concentration of PTS was 200 micrograms/ml, CFU-E and BFU-E was zero respectively. In the CFU-GM culture, PTS in concentration of 12.5 micrograms/ml could cause the proliferation increased by 29.7 +/- 2.2% (P < 0.05), but in concentration of 100 micrograms/ml and 200 micrograms/ml, it showed inhibitory effect on CFU-GM, the inhibition rate being 48.6 +/- 3.9% and 100% respectively. CONCLUSION: PDS is the effective component of ginsenosides in stimulating proliferation of human bone marrow HPC. PTS is an component with inhibitory action on proliferation of CFU-E and BFU-E and its effect on CFU-GM was depending on its concentration.

[Proliferation and differentiation of human CD34+ hematopoietic stem/progenitor cells induced by Panax notoginosides].

The object of this study was to explore the effects of Panax notoginosides (PNS) on proliferation and differentiation of human CD34(+) stem/progenitor cells. CD34(+) cells were isolated from human bone marrow by using immune beads of Dynal M- 450 system. The cells were exposed to PNS at different concentrations in both liquid and semi-solid culture for 14 days. The cells were marked with monoclonal antibodies and analyzed by flow cytometry after culture. The CFU-Mix colony formation from CD34(+) cells was assayed. The results showed that: (1) The yield of CD34(+) cells after being selected by immune beads were (1.03 +/- 0.74)% out of bone marrow nuclear cells with purity of 86% - 93%. (2) PNS (10 - 25 mg/L) stimulated the proliferation of CD34(+) cells, and raised the colony numbers of CFU-Mix obviously in vitro. PNS 25 mg/L was the optimal concentration to promote proliferation of CD34(+) cells, the increasing rate of CFU-Mix colony was (34.7 +/- 16.0)%. (3) The differentiation of CD34(+) cells was induced by exposure to PNS (25, 50 and 100 mg/L) in liquid culture for 14 days. The percentages of CD33(+) and CD15(+) cells were increased after PNS exposure, which were significantly higher than those of control (P < 0.01), however CD71(+) and G-A(+) cells were no obviously difference after PNS treatment. In conclusion, Panax notoginosides not only promote the proliferation of CD34(+) cells, but also induce the differentiation committed to granulocytes.