In this paper, the varying pattern of the amount of rhizospheric microorganisms, including bacteria, actinomycetes and fungus, was observed during the cultivation of Paris polyphylla var. yunnanensis. And the correlations between number of rhizospheric microorganisms and the quality of P. polyphylla var. yunnanensis were also studied. The results showed that the rhizospheric microorganism source of P. polyphylla var. yunnanensis was rich. The distribution of rhizospheric microorganisms (soil bacteria, fungus, actinomycetes, potassium-solubilizing bacteria, inorganic phosphorus-solubilizing bacteria, organic phosphorus-solubilizing bacteria) collected from different origin places existed significant difference (P < 0.05). The varying pattern for the amount of rhizospheric microorganisms was showed as following: the amount of bacteria > the amount of actinomycetes > the amount of fungus. The medicinal quality of P. polyphylla var. yunnanensis was influenced by their habits, and the increase of cultivation years caused the obvious decrease of the quality of P. polyphylla var. yunnanensis. Therefore, the increase of cultivation years will cause the variation of the soil micro-ecology flora, and decrease the nutrient absorption and the utilization of P. polyphylla var. yunnanensis, which will make the decrease of the medical quality of P. polyphylla var. yunnanensis.
Bacteria ; genetics ; growth & development ; isolation & purification ; Biodiversity ; China ; Fungi ; genetics ; growth & development ; isolation & purification ; Liliaceae ; chemistry ; microbiology ; Plant Extracts ; analysis ; Rhizome ; chemistry ; microbiology ; Rhizosphere ; Saponins ; analysis ; Soil Microbiology

AIM: To purify human yolk sac mesenchymal ste m cells (hYS-MSC) and investigate its osteogenic and neurogenic differentiation potentials. METHODS: hYS-MSC were separated from yolk sac and purified via p assage culture. The karyotype of hYS-MSCs was analyzed via G-banded characterist ics. Flow cytometric analysis was used to determine the cell cycle and phenotype of hYS-MSC. The AKP expression of hYS-MSC was also tested. Osteogenic different iation of hYS-MSCs was induced by 10 -8mol/L dexamethasone, 10 mmol/L ?-gl ycerophosphate and 50 mg/L vitamin C. Alizarin red S stain was used for identifi cation of mineralization. ?-mecaptoethanol or salviae miltiorrhizae were used t o induce neurogenic differentiation of hYS-MSCs. The expressions of NSE, NF and GFAP were identified by immunohistochemical method. RESULTS: hYS-MSCs could be purified at passages 2 or 3. The cell cycle analysis suggested that hYS-MSCs showed strong proliferational potentials by which the cells kept normal diploid karyotype during the in vitro cultur e. Flow cytometry showed the phenotype of purified hYS-MSCs was uniformly positi ve for CD29, CD44, CD105, and CD166, and negative for reactivity to antigens CD3 4, CD45, or CD86. hYS-MSCs were weakly but clearly positive in AKP. Osteogenic d ifferentiation was appeared after induction of osteogenic differentiation. hYS-M SCs, which were of spindle shape, uniform in size, were induced to pleomorphism osteoblast-like cells which expressed high level of AKP. Aggregates or nodules w ere formed at day 7 and calcium accumulation was detected by alizarin red S stai ning on day 10 or day 14. Neurogenic differentiation of hYS-MSCs was induced by ?-mecaptoethanol or salviae miltiorrhizae. NSE, NF or GFAP positive cells were detected by immunohistochemical staining. CONCLUSIONS: hYS-MSCs have strong proliferation potential and th e normal diploid karyotype is kept during the in vitro culture. The phenoty pe of hYS-MSCs is coincident with adult hMSCs. hYS-MSCs could be induced to dif ferent iate into osteogenic or neurogenic cells.

OBJECTIVETo introduce the newly found gene TIP30/CC3 into a hepatoma cell line PLC/PRF/5 and select the stable expression clones. The growth and cell cycles were studied with the clones stably expressing TIP30/CC3 or anti-TIP30/CC3, and the effects of TIP30/CC3 gene on hepatoma cells were analyzed.

METHODSThe internal expression of TIP30/CC3 protein was detected with Western blot, then TIP30/CC3 or anti-TIP30/CC3 cDNA was subcloned into a constitutive vector pcDNA3 followed by transfection into PLC/PRF/5. Stable expression clones were selected. The cell growth curve was made and cell cycles detected using flow cytometry. To confirm the results in vitro, stable-expressing cells were implanted subcutaneously into nude mice and time of tumor formation recorded and tumor volume measured.

RESULTSPLC-anti-TIP30 grew faster than the others. Three days after transfection, live cells of PLC-anti-TIP30 were 14.0*10(4), in comparison with the control PLC-DNA3 and PLC/PRF/5, the differences were statistically significant. Live cells of PLC-TIP30 were 4.9*10(4), significantly less than the two control groups. Six days after transfection, live cells of PLC-anti-TIP30 were 25.0*10(4), significantly more than the controls PLC-DNA3 and PLC/PRF/5. Live cells of PLC-TIP30 were 12.4*10(4), significantly less than the two control groups. Cell cycle analysis showed that PLC-anti-TIP30 proliferated faster, 22.4% cells were in G0/G1 (gap) phases and 58.6% cells in S (DNA synthesis) phase. The growth of the PLC-anti-TIP30 cell was retarded and many cells were arrested from G1 to S phases. Cells in G0/G1 and S phase were 44.2% and 33.3% respectively. Furthermore, the average time of tumor formation was shorter in anti-TIP30 group and longer in TIP30/CC3 group, and times were 6.0 d (with control groups) and 15.6 d (with control groups) respectively. Tumors in the nude mice grew faster in PLC-anti-TIP30 group and slower in PLC-TIP30 group.

CONCLUSIONTumor suppressor gene TIP30/CC3 can inhibit the proliferation of tumor cells and interfere in its cell cycles. It can be used as a valuable tool for hepatoma biotherapy including gene therapy.

Acetyltransferases ; biosynthesis ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genes, Tumor Suppressor ; Humans ; Liver Neoplasms ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Transcription Factors ; biosynthesis ; genetics ; Transfection

To investigate the pathophysiological mechanisms for platelet-neutrophils cross talk mediated by platelet-activating factor (PAF) and to lay a foundation for clinical application, ginkgolides B (GB), a PAF receptor antagonist, was added in the whole blood to block the effects of PAF on activation of platelet-neutrophil; PAF and ADP were respectively added in the whole blood to monitor the expression of CD62P on platelet by flow cytometry; PAF and ADP were added in the whole blood to monitor the expression of CD11b on neutrophil by flow cytometry; PAF and ADP were added in the whole blood to monitor the platelet-leucocyte aggregates (PLA) which were PLA in the total leucocyte population (PLA/L) and the mean fluorescence intensity (MFI) of CD42b. Outcomes were analyzed by t-test, and the differences were statistically significant (P < 0.05). The results showed that the expression of CD62P on platelats, the expression of CD11b on neutrophils and PLA formation were all increased by PAF and ADP; the PAF receptor antagonists (GB) could obviously inhibit the expression of CD62P, CD11b and PLA formation induced by PAF, but could not completely inhibit the activation of platelet and neutrophil, and the platelet-neutrophil cross talk; GB could inhibit the expression of CD62P and CD11b induced by ADP, but could not conpletely inhibit the activation of platelet and neutrophli; GB could not obviously inhibit the platelet-leucocyte aggregates mediated by ADP. It is concluded that the multiligand-receptor systems involved in PLA formation and platelet-netrophils cross talk seem to be regulated by complex mechanisms; the PAF receptor antagonists (GB) obviously inhibit the effect of PAF, and may be widely utilized in the therapy of thrombosis and inflammation.
Adenosine Diphosphate ; pharmacology ; Adult ; Blood Platelets ; cytology ; drug effects ; physiology ; CD11b Antigen ; blood ; Cell Adhesion ; drug effects ; physiology ; Cell Communication ; drug effects ; physiology ; Female ; Flow Cytometry ; Ginkgolides ; pharmacology ; Humans ; Male ; Middle Aged ; Neutrophils ; cytology ; drug effects ; physiology ; P-Selectin ; blood ; Platelet Activating Factor ; pharmacology ; Platelet Activation ; drug effects ; Platelet Glycoprotein GPIb-IX Complex ; analysis ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; physiology ; Receptors, G-Protein-Coupled ; antagonists & inhibitors ; physiology

Objective To establish a model of acoustically evoked short latency negative response (ASNR) in guinea pigs, a model of profound hearing loss with normal saccular functions, and verify the correlation between ASNR and vestibular evoked myogenic potential (VEMP). Methods Thirty-two healthy guinea pigs were employed in the experiment, which were randomly divided into control group ( 16subjects) and deafened group (16 subjects). Each animal experienced auditory and vestibular tests including auditory brainstem response ( ABR), VEMP and caloric test. A quick treatment was employed for deafened group consisting of a subcutaneous injection of kanamycin at a dose of 400 mg/kg followed by a jugular vein injection of ethacrynic acid at a dose of 40 mg/kg one hour later. The animals were received ABR, VEMP and caloric test 7 - 10 days following the drug administration. The deafened group was further divided into ASNR group and non-ASNR group, based on the presence of ASNR. Results In deafened group, five subjects died postoperatively, 11 subjects (22 ears) provided full data, ASNR was elicited in eight ears (36.4%), the threshold was 120- 130 dB SPL with mean of (124.4 ±4.96) dB SPL. Its latency range was 1.75 - 2. 60 ms with mean of ( 2. 15 ± 0. 27 ) ms. The mean latency of threshold was (2. 34 ±0. 18) ms. All eight ASNR ears presented with VEMP. The VEMP threshold, positive and negative potential latencies proved no statistical difference (P ＞ 0. 05 ) between ASNR group and control group.Significant difference was detected between the VEMP presence of ASNR group and non-ASNR group ( P =0. 002). There was no statistically significant correlation between VEMP and caloric test neither between ASNR and caloric test in deafened group. Conclusions This study evoked ASNR in an ototoxicity guinea pig model which has profound hearing loss with normal saccular functions. The presence of ASNR correlated with VEMP, however, not correlated with caloric test, suggesting that ASNR and VEMP are both originated from the saccule.

OBJECTIVETo investigate the effect of phenylhexyl isothiocyanate (PHI) on histone acetylation and apoptosis in hepatocellular carcinoma cell line (SMMC-7721) in vitro.

METHODSThe viability of SMMC-7721 cells was determined by trypan blue exclusion. Apoptotic cells were assessed by TUNEL assay. The proteins of Bcl-2, Procaspase-9, Procaspase-8, Procaspase-3, caspase-9, caspase-3, histone acetylated H3 and H4 were detected by Western blot.

RESULTSCompared with the vehicle control, PHI at 5, 10, 20, 40 and 80 µmol/L reduced the cell viability of SMMC-7721 cells in a concentration-dependent manner. PHI induced apoptosis in SMMC-7721 cells. An increased amount of apoptotic cells was detected after 7 hours exposure to PHI at 10, 20, and 40 µmol/L, 6.9% ± 2.4%, 17.5% ± 4.2% and 54.5% ± 5.4%, respectively, while that of the vehicle control was 4.5% ± 2.3% (P < 0.05). Along with the prolongation of time and increase of dose, the expressions of bcl-2, procaspase-9, procaspase-3 were decreased, that of caspase-9 and caspase-3 was increased. In contrast, alteration of procaspase-8 was not significant at those concentrations. PHI accumulated acetylated histone H3 and H4. After 3 hours exposure to PHI at 10, 20 and 40 µmol/L, the level of histone acetylated H3 was 1.87-, 2.43-, 3.67-fold increased and histone acetylated H4 was 1.29-, 1.45-, and 2.25-fold increased, compared with that of the vehicle control. The protein of histone acetylated H3 and H4 was significantly accumulated after 7 hours exposure.

CONCLUSIONPHI is a new histone deacetylation inhibitor. It may induce accumulation of histone acetylation H3 and H4, inhibit cell growth and induce apoptosis in SMMC-7721 cells via the mitochondrial pathway.

Acetylation ; drug effects ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Histone Deacetylase Inhibitors ; administration & dosage ; pharmacology ; Histones ; metabolism ; Humans ; Isothiocyanates ; administration & dosage ; pharmacology ; Liver Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

OBJECTIVETo evaluate the potential usefulness of a multivariable model in predicting the response to immunosuppressive therapy (IST) in patients with aplastic anemia (AA), and its application to the clinical practice.

METHODSPB T cells subpopulation and BM T cells intracellular IFN-gamma and IL-4 were serially analyzed by flow cytometry (FCM) before and during treatment. HLA-DRB1 * 1501 phenotype was analyzed by PCR-SSP. The predictive potentials of different parameter combinations for clinical responsiveness were statistically assessed.

RESULTSIn all evaluated parameters, CD8+ cell intracellular IFN-gamma had the relatively best diagnostic value with sensitivity and specificity of 94.3% and 62.5%, and positive and negative predictive value of 84.6% and 83.3% respectively. Positive CD8+ cell intracellular IFN-gamma plus Tc1/Tc2 < 50 could increase the positive predictive value to 92.3%. A multivariable model consisting of absolute neutrophil count (ANC), BM T cell intracellular IFN-gamma, Tc1/Tc2 ratio and HLA-DRB * 1501 phenotype of the patients was finally established.

CONCLUSIONThe multivariable model is superior to each of the single parameters in terms of predictive power of IST therapeutic outcome, and its higher accuracy and the clinical application make it potentially useful in practice.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; drug therapy ; immunology ; Child ; Feasibility Studies ; Female ; HLA-DR Antigens ; immunology ; Humans ; Immunosuppression ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Models, Statistical ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology ; Treatment Outcome

OBJECTIVETo compare therapeutic effects of warm needle moxibustion and medication on osteoporosis and to study the mechanism.

METHODSForty cases were randomly divided into an acupuncture group and a medication group, 20 cases in each group. The acupuncture group was treated by warm needle moxibustion at Dazhu (BL 11), Ganshu (BL 18), Shenshu (BL 23), Zusanli (ST 36), Yanglingquan (GB 34) etc. once other day, for 3 months; and the medication group was treated by oral administration of tablet Caltrate with Vit D2 for 3 months. The changes of bone mass density (BMD), estradiol (E2), osteocalcin (bone growth protein, BGP), urine calcium/creatinine (Ca/Cr) in the two groups before and after treatment and therapeutic effects were investigated.

RESULTSAfter treatment, BMD significantly increased (P<0.05, P<0.01) in the acupuncture group and did not signifi cantly changed in the medication group (P>0.05) with a significant difference between the two groups (P<0.05). After treatment E2 level significantly increased as compared with before treatment in both of groups (P<0.01); after treatment BGP significantly decreased as compared with before treatment in both of groups (P<0.01); after treatment Ca/Cr significantly decreased as compared with before treatment in the acupuncture group (P<0.05) ; af ter treatment, there were significant differences in BGP and Ca/Cr between the two groups (P<0.05 or P<0.01). The clinically controlled rate in the acupuncture group and in the medication group were 35.0%, 5.0%, respectively, the therapeutic effect of the acupuncture group being better than that of the medication group (P<0.01).

CONCLUSIONThe therapeutic effect of warm needle moxibustion on osteoporosis is better than that of oral administration of tablet Caltrate with Vit D2 and it can increase levels of hormones and delay bone loss. It is an effective method for preventing and treating postmenopausal osteoporosis.

Acupuncture Points ; Aged ; Bone Density ; Bone and Bones ; metabolism ; physiopathology ; Calcium ; urine ; Creatinine ; urine ; Estradiol ; metabolism ; Female ; Humans ; Middle Aged ; Moxibustion ; Osteocalcin ; metabolism ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; therapy

OBJECTIVETo investigate the expression of AML1/ETO9a isoform in the acute myeloid leukemia (AML)-M2 patients.

METHODSExpressions of AML1/ETO fusion gene and AML1/ETO9a isoform were detected by using reverse transcriptase-polymerase chain reaction (RT-PCR) in leukemia patients, MDS patients, leukemia cell lines and healthy subjects. Karyotype was studied by R-banding technique.

RESULTIn 30 newly diagnosed AML-M2 patients 15 were found to express AML1/ETO9a isoform, while the rest including 20 AML-M2CR, 18 other subtypes of AML, 5 chronic myelogenous leukemia (CML), 3 myelodysplastic syndromes (MDS), 3 leukemia cell lines (NB4, KG-1, K562) and 5 healthy subjects were AML1/ETO9a negative. Among the 15 AML/ETO9a isoform expressing cases, 13 were demonstrated t(8;21) translocation and AML1/ETO expression.

CONCLUSIONIsoform AML1/ETO9a was correlated to AML/M2, and it may promote the development of leukemia in combination with the AML1/ETO fusion gene.

Adolescent ; Adult ; Aged ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Female ; Gene Expression ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Protein Isoforms ; genetics ; metabolism ; RUNX1 Translocation Partner 1 Protein

OBJECTIVETo evaluate whether neutrophil-lymphocyte ratio(NLR) predicts risk of recurrence in patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy.

METHODSA total of 149 patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy(FOLFOX6 protocol) were included. NLR was calculated preoperatively and before chemotherapy. The changes in NLR and the predictive value of NLR for prognosis were analyzed.

RESULTSThe NLR of 149 patients was 2.8±1.5. NLR of 3.5 was identified according to the ROC curve. NLR<3.5 and NLR≥3.5 were classified as low and high NLR group, respectively. The 5-year recurrence-free survival(RFS) of patients with high preoperative NLR(n=22) was significantly worse than that of those with low preoperative NLR(n=127)(50.9% vs. 76.4%, P=0.025). The difference of 5-year RFS between high pre-chemotherapy NLR group(n=34) and low pre-chemotherapy NLR group(n=115) was statistically significant(50.1% vs. 71.4%, P=0.032). The 5-year RFS was 79.5% in patients with low preoperative NLR converting to high pre-chemotherapy NLR(n=16), similar to the group with high pre-chemotherapy group(P=0.077). The 5-year RFS was 17.7% in patients with high preoperative NLR reverting to low pre-chemotherapy NLR(n=12), similar to the group with low pre-chemotherapy group(P=0.978). There was significant difference in 5-year RFS between the postoperatively elevated group and postoperatively decreased group(P=0.036).

CONCLUSIONAn elevated blood NLR may be a biomarker of poor RFS in patients with advanced colon cancer after curative resection and chemotherapy.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Chemotherapy, Adjuvant ; Colonic Neoplasms ; blood ; therapy ; Disease-Free Survival ; Female ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Lymphocytes ; immunology ; Male ; Middle Aged ; Neutrophils ; immunology ; Organoplatinum Compounds ; therapeutic use ; Prognosis