Effects of LW-AFC, a new formula derived from Liuwei Dihuang decoction, on intestinal microbiome in senescence-accelerated mouse prone 8 strain, a mouse model of Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effects of LW-AFC, a new formula derived from Liuwei Dihuang decoction, on gut microbiota and the behavior of learning and memory of SAMP8 mice, a mouse model of Alzheimer Disease (AD), and identify the specific intestinal microbiota correlating with cognitive ability. METHODS Morris-water maze test, novel object recognition test and shuttle-box test were conducted to observe the ability of learning and memory. 16S rRNA amplicon sequencing (Illumina, San Diego, CA, USA) was employed to investigate gut microbiota. RESULTS The treatment of LW- AFC improved cognitive impairments of SAMP8 mice, including spatial learning and memory ability, active avoidance response, and object recognition memory capability. Our data indicated that there were significantly 8 increased and 12 decreased operational taxonomic units (OTUs) in the gut microbiota of SAMP8 mice compared with senescence accelerated mouse resistant 1 (SAMR1) strains, the control of SAMP8 mice. The treatment of LW- AFC altered 22 (16 increased and 6 decreased) OTUs in SAMP8 mice and among them, 15 OTUs could be reversed by LW-AFC treatment resulting in a microbial composition similar to that of SAMR1 mice. We further showed that there were 7 (3 negative and 4 positive correlation) OTUs significantly correlated with all the three types of cognitive abilities, at the order level, including Bacteroidales, Clostridiales, Desulfovibrionales, CW040, and two unclassified orders. LW-AFC had influences on bacterial taxa correlated with the abilities of learning and memory in SAMP8 mice and restored them to SAMR1 mice. CONCLUSION The effects of LW-AFC on improving cognitive impairments of SAMP8 mice might be via modulating intestinal microbiome and LW-AFC could be used as a potential anti-AD agent.

Performance evaluation of DIRUI FUS-2000 automated urine analyzer for detecting urinary RBC and WBC / 医疗卫生装备

Objective To validate the performance parameters of DIRUI FUS-2000 automated urine analyzer declared by the manufacturer.Methods The fresh urine samples were diluted with special diluent to different concentration of suspension according to experimental requirements,then such parameters were validated as the precision,carrying pollution rate,detection limit and reportable range.At the same time,the coincidence rate of red blood cells and white blood cells was also verified in urine samples tested by FUS-2000 according to the result of manual inspection.Results RBC had the low and median values for within-run precision being 7.74％ and 6.13％ respectively,and WBC had the low and median values for within-run precision being 14.35％ and 2.45％ respectively.The negative and positive quality control materials had the between-run precision being 0％ and 3.43％ respectively.The carrying pollution rate was 0.08％ for RBC and 0.57％ for WBC,and the detection limits for RBC and WBC were both 10 ones per Liter.The reportable range was from 10 to 38 754 p/μl for RBC and from 10 to 32 202 p/μl for WBC.The coincidence rate with artificial microscopy was 90.83％ for RBC and 95.83％ for WBC.Conclusion FUS-2000's performance parameters are in line with the requirement,and it can be used for the clinical urine detection.The review criteria of FUS-2000 should be established to improve distinguishing sensitivity and specificity of RBC and WBC in urine.

Performance evaluation of DIRUI FUS-2000 automated urine analyzer for detecting urinary RBC and WBC / 医疗卫生装备

Objective To validate the performance parameters of DIRUI FUS-2000 automated urine analyzer declared by the manufacturer.Methods The fresh urine samples were diluted with special diluent to different concentration of suspension according to experimental requirements,then such parameters were validated as the precision,carrying pollution rate,detection limit and reportable range.At the same time,the coincidence rate of red blood cells and white blood cells was also verified in urine samples tested by FUS-2000 according to the result of manual inspection.Results RBC had the low and median values for within-run precision being 7.74％ and 6.13％ respectively,and WBC had the low and median values for within-run precision being 14.35％ and 2.45％ respectively.The negative and positive quality control materials had the between-run precision being 0％ and 3.43％ respectively.The carrying pollution rate was 0.08％ for RBC and 0.57％ for WBC,and the detection limits for RBC and WBC were both 10 ones per Liter.The reportable range was from 10 to 38 754 p/μl for RBC and from 10 to 32 202 p/μl for WBC.The coincidence rate with artificial microscopy was 90.83％ for RBC and 95.83％ for WBC.Conclusion FUS-2000's performance parameters are in line with the requirement,and it can be used for the clinical urine detection.The review criteria of FUS-2000 should be established to improve distinguishing sensitivity and specificity of RBC and WBC in urine.

Selection and Identification of the Biological Characteristics of a Cold-adapted Genotype G1P8 ZTR-68 Rotavirus by Serial Cold-adapted Passaging / 病毒学报

We wished to select a cold-adapted genotype G1P[8] ZTR-68 rotavirus (China southwest strain) in MA104 cells for possible use as a live vaccine. ZTR-68 was recovered originally from children with diarrhea. The virus was cultivated at 37 degrees C at the first passage. Then, the cultivation temperature was decreased stepwise by 3 degrees C per eight passages. In total, the virus was passaged 32 times, and cultivation was terminated at 28 degrees C. Biological characteristics of the virus were analyzed during serial passages. There was no difference between the migration patterns of genomic dsRNA segments according to polyacrylamide gel electrophoresis of original and cold-adapted viruses. Infectious and red cell-agglutination titers of cold-adapted virus were lower than those of the parent virus. Also, the virus formed small-size plaques with irregular shapes at 31 degrees C and 28 degrees C. These results suggested that a genetically stable attenuated virus can be obtained through serial cold-adapted passages. Thus, an alternative strategy is provided by cold-adaption for development of attenuated live rotavirus vaccines.

Establishment and Applications of Double-Fluorescent Protein Allo-Transplantation Mice Model / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish allo-transplantation model by using mRFP⁺ to eGFP⁺ transgenic mice and to observe the distribution of donor cells and donor-recipient cellular interaction in the bone marrow after semi-solid decalcification (SSD).</p><p><b>METHODS</b>After myeloablative irradiation, C57BL/6 female eGFP⁺ transgenic mice were infused with (5 × 10⁶) bone marrow cells from FVB male donor mice through tail vein. The control group was infused with PBS. Then the general conditions, engraftment level, hematopoietic recovery, incidence of GVHD and survival of recipients were evaluated after transplantation. In the recovery process, SSD was used to treat the femora before observing the cells distribution, morphology and interaction by confocal microscopy directly or after making frozen section.</p><p><b>RESULTS</b>WBC of recipient eGFP⁺ mice was recovered on (20 ± 3.07) d, (93.94 ± 1.59)% in peripheral cells were RFP⁺ cells (n = 10), GVHD happened in 4 of 10 mice within 1 month. During SSD, the hard components were replaced gradually and RFP⁺ cells could be seen mainly in the bone trabecula and surrounded by eGFP⁺ cells under confocal microscope, their interactions could be further observed clearly in bone marrow microenvironment in three-dimensional reconstruction.</p><p><b>CONCLUSION</b>The double fluorescent allo-transplantation mouse model successfully established, by means of our novel protocol named SSD, the donor and recipient cell location and their interaction can be visually observed, which provides the basis for clinical studies on the distribution and homing of donor cells, and some related explorations after transplantation.</p>

HPLC fingerprint of glycyrrhizea radix et rhizoma praeparata cum melle / 中国中药杂志

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.

Anti-apoptotic effect of Astragalus Polysaccharide on myeloid cells / 中国实验血液学杂志

This study was aimed to assess the effect of Astragalus Polysaccharide (ASPS) on in-vitro hematopoiesis. CFU-GM assays were used to determine the effect of ASPS and thrombopoietin (TPO) on granulocytic-monocyte progenitor cells. The CFU assays were also used to investigate the effect of ASPS on the proliferation of HL-60 cells.HL-60 cells were cultured with serum-free RPMI 1640 medium and treated with or without of different concentrations of ASPS. After 72 h incubation, the number of cells were counted.In addition, the caspase-3 and JC-1 expression was determined by flow cytometry with Annexin V/PI double staining. The results showed that ASPS (100, 200 µg/ml) and TPO (100 ng/ml) significantly promoted CFU-GM formation in vitro. Various concentrations of ASPS and TPO also promoted the colony formation of HL-60 cells, the largest effect of ASPS was observed at a concentration of 100 µg/ml. There were no synergistic effects between TPO and ASPS on cellular proliferation. The results also showed that ASPS significantly protected HL-60 cells from apoptosis in condition of serum-free medium culture, suppressed caspase 3 activation, and reduced the cell apoptosis. It is concluded that ASPS can significantly promote the formation of bone marrow CFU-GM and the proliferation of HL-60 cells, the optimal concentration of ASPS is at 100 µg/ml. In the absence of serum inducing apoptosis, ASPS also significantly reduced the apoptosis of HL-60 cells via suppressing the activation of caspase-3.

Application of quantum dots labeling technique in induced pluripotent stem cells / 中华实验眼科杂志

Background The multipotent differentiation features of induced pluripotent stem cells (iPSCs) offer a new option for cell replacement therapy of many clinical diseases.In ophthalmology,iPSCs are a good model in studying the pathogenic mechanism of degenerative ocular diseases.A better identification method for iPSCs is critical for analyzing the in vivo biological characteristics of iPSCs.Objective This study was to investigate the feasibility and stability of labeling iPSCs with quantum dots.Methods Human umbilical mesenchymal stromal cells-iPSC lines were cultured and amplified on matrigel,and the characteristics of iPSCs were evaluated by immunofluorescence.Different concentrations (5.0,7.5 and 10.0 nmol/L) of quantum dots with a CdSe/ZnS nuclear shell structure were used to label iPSCs after passaging and proliferation.The labeling outcome was observed with a three-dimensional deconvolution real-time live cells imaging system.The labeled iPSCs were subsequently cultivated,and then changes in fluorescence intensity were examined 7 days after the first and the second passaging of iPSCs.Results iPSCs were observed to grow in a clonal manner under the inverted microscope.The iPSC markers,OCT4 and Nanog,were detected by immunofluorescence.With increasing concentrations of quantum dots,the fluorescence intensities representing the levels of OCT4 and Nanog in iPSCs were gradually elevated,with optimal levels of fluorescence observed at a concentration of 10 nmol/L of quantum dots.The fluorescent labeling of OCT4 and Nanog in iPSCs remained and weakened gradually till day 7 even after the second passage.Conclusions Quantum dots labeling could be used to track iPSCs in a dose-independent manner.The fluorescent signal from the quantum dots labeling the iPSCs lasts 2 weeks at least.

Influence of aqueous humor on growth of bovine corneal endothelial cell in vitro / 中华实验眼科杂志

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

HPLC fingerprint of Liuwei Dihuang condensed pills / 中国中药杂志

<p><b>OBJECTIVE</b>To establish HPLC fingerprints of Liuwei Dihuang condensed pills.</p><p><b>METHOD</b>Dikma Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with acetonitrile (containing 0.05% phosphoric) -water (containing 0.05% phosphoric) as the mobile phase. The column temperature was set at 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was 276 nm (0-10 min), 236 nm (10-40 min) and 276 nm (40-60 min). The sample size was 20 microL. Chromatographic peaks were identified by Q-TOF-MS-IDA-MS/MS method.</p><p><b>RESULT</b>Good precision, stability and repeatability were proved. Q-TOF-MS-IDA-MS/ MS method was adopted for qualitative determination of eighteen chromatographic peaks. Ten batches of Liuwei Dihuang condensed pills were determined with the method, and their similarities were above 0. 96.</p><p><b>CONCLUSION</b>The study lays a foundation for the overall quality evaluation of Liuwei Dihuang condensed pills.</p>

Chemical constituents of glucoside fraction from Liuwei Dihuang Gantang / 中国中药杂志

<p><b>OBJECTIVE</b>To study chemical constituents of glucoside fraction from Liuwei Dihuang Gantang and clarify its substance foundation of active constituents.</p><p><b>METHOD</b>Glucoside fraction was prepared by macroporous resin chromatography. Constituents were separated by silica gel and reverse phase silica gel column chromatography, and their structures were identified by MS and NMR.</p><p><b>RESULT</b>Eleven compounds were separated and identified as 7-dehydrologanin (1), 7alpha-O-methylmorroniside (2), 7beta-O-methylmorroniside (3), 7alpha-O-ethylmorroniside (4), 7beta-O-ethylmorroniside (5), morroniside (6), sweroside (7), loganin (8), paeoniflorin (9), paeonolide (10) and loganic acid (11).</p><p><b>CONCLUSION</b>All of those compounds were separated from Liuwei Dihuang Gantang for the first time.</p>

The evaluation of immune effects on bivalent rotavirus vaccine / 中华微生物学和免疫学杂志

Objective To evaluate the immune effects of bivalent inactivated rotavirus vaccine (IRV) and investigate the viability of development of bivalent IRV.Methods Firstly,bivalent IRV was prepared by mixing G1 IRV and G3 IRV with equal amount,G1 IRV and G3 IRV as monovalent control,PBS as negative control.Secondly,those vaccines were vaccinated to the mice by intramuscular injection.Then,to evaluate the immune effects of bivalent IRV,the levels of serum or fecal rotavirus specific IgG and IgA were assessed by ELISA,the levels of serum neutralized antibody were measured by microneutralization assay,the number of IFN-γ or IL-4 secreting cells were analyzed by ELISPOT assay.Results Compared to negative control group,bivalent IRV induced the higher levels of serum and fecal G1 and G3 rotavirus specific antibody.It was found that there were no significant differences for the levels of serum IgG and IgA,fecal IgG and IgA,serum neutralized antibody between induced by bivalent IRV and induced by G1 type monovalent vaccines ; but there were significantly increase for the levels of serum IgG (t =2.691,P＜0.05) and serum neutralized antibody (t =2.561,P＜0.05) between induced by bivalent IRV and induced by G3 monovalent vaccines,there were no significant differences for other antibodies between induced by bivalent IRV and induced by G3 monovalent vaccines.At the same time,compared to negative control group,bivalent IRV induced significantly increase in the number of IFN-γ or IL-4 secreting cells in spleen lymphocytes.It was found that there were no significant differences for the number of IFN-γ or IL-4 secreting cells stimulated by G1 rotavirus between bivalent IRV and G1 monovalent vaccines; but there were significantly increase for the number of IL-4 secreting cells (t =2.327,P＜0.05) stimulated by G3 rotavirus between bivalent IRV and G3 monovalent vaccines,there were no significant differences for the number of IFN-γ secreting cells stimulated by G3 rotavirus between bivalent IRV and G3 type monovalent vaccines.Conclusion The bivalent IRV can induce effective immune response,in which there were no inhibitory interference between the components of bivalent IRV,which provided the experimental basis for the development of bivalent IRV.

Morphologic observation of induced pluripotent stem cells induced by corneal endothelium cells with atomic force microscopy / 中华实验眼科杂志

Background Induced pluripotent stem cells (iPSCs)can differentiate into various types of somatic cells without causing ethical controversy and immune rejection in clinical activity,which is similar to differentiation ability of embryonic stem cells.So,iPSCs may be used as seed cells for tissue engineering corneal endothelial reconstruction.Objective The present study was to survey the morphologic change of iPSCs after coculture with corneal endothelium cells(CECs) under the atomic force microscopy(AFM).Methods Rabbit CECs and human MMC-iPSCs were isolated and cultured respectively.The iPSCs were identified with the marker by immunochemistry.iPSCs passaged for 7 days were then cultured with 60％ confluent CECs to establish the co-culture model.The surface morphology and cellular membrane ultrastructure of differentiated iPSCs after induced by CECs were examined by AFM combination with inverted microscope,and compared with CECs and undifferentiated iPSCs.Results Thelengthand width were(66.93±10.48)μm and (44.85 ± 8.14) μm in CECs,(12.51±1.40)μm and (10.93 ±1.69) μm in uninduced iPSCs,and(36.12±10.29) μm and(31.53±9.65)μm in CECs-induced iPSCs.Both the length and width values of CECs-induced iPSCs were statistically bigger than those uninduced iPSCs,with significant differences between them (P＜0.05),but no significant difference was seen in the width valne of CECs-induced iPSCs in comparison with CECs(P＞0.05).The convex structure of CECs cytomembrane surface showed the digitation in shape with the size and height(2.11 ± 1.03) μm and (115.68±92.08) nm respectively,and the concave structure of cytomembrane surface of CECs was fenestrae-like depression and the size was (1.49 ± 0.65) μm.The numerical valuc of mean square root roughness (Rq)and average roughness (Ra)of cytomembrane surface of CECs were(39.20±7.82)nm and (30.37±5.32)nm respectively.The convex surface of cytomembrane of iPSCs was granular-like in shape with size and height(0.39±0.22)μm and(13.11±9.18)nm respectively.The concave surface of cytomembrane of iPSCs was worm-eaten-like concave with the size(0.34±0.18)μm.The numerical value of Rq and Ra of geometrical parameters of cytomembrane surface of iPSCs were (26.60 ± 4.93)nm and (9.97 ± 3.78) nm respectively.The convex surface of cytomembrane of induced iPSCs was digital-like in shape with the size and height (1.91±0.76) μm and(106.55±77.27) nm respectively.The concave surface of cytomembrane of induced iPSCs was fenestrae-like depression and the size of concave was(1.6l±1.25) μm.The numerical value of Rq and Ra on surface of cytomembrane of induced iPSCs was (57.33± 12.80) nm and (43.63± 11.17) nm respectively.The numerical values of the size and height of convex,the size of concave,Rq and Ra on surface of cytomembrane in induced iPSCs were statistically bigger than in iPSCs(P＜0.05)and were not significant differences in comparison with CECs (P＞0.05).Conclusions Morphology of iPSCs translate toward the CECs after induce for 7 days under the AFM.This outcome lays the foundation for further study on iPSCs.

Simultaneous analysis of telomere length and cell surface antigen in leukemia by multicolor Flow-FISH / 中国实验血液学杂志

This study was purposed to explore the feasibility of simultaneous analysis of telomere length and cell surface antigen by multicolor Flow-FISH to assess minimal residual disease (MRD) in leukemia. The telomere length in 34 leukemia patients versus 20 normal controls was compared by using Flow-FISH, and the relationship between telomere length and therapeutic effect and prognosis was analyzed preliminarily. As for those patients with follow-up samples, the changes of telomere length combined with surface antigen in different courses of disease were observed by multicolor Flow-FISH. The results indicated that the telomere length of de novo patients was significantly shorter than that of controls except the patients in chronic myeloid leukemia-chronic phase (CML-CP). The shorter telomere, the lower complete remission (CR) rates were observed in acute leukemia cases and the shorter duration of CP before onset of blast phase (BP) occurred in CML cases. The acute leukemia patients showed longer telomere and fewer cells expressed the related antigen after CR. The telomere length of cases with continued CR remained at normal level during remission, and there was no increased expression of the specific antigen. However, the telomere of relapsed cases shortened again after relapse with elevated specific antigen expression. In the relapsed cases, the telomere of related antigen positive cells shortened ahead of telomere length change of the whole cells and morphologic change of bone marrow cells. It is concluded that analysis of telomere length by flow-FISH manifests the significance for monitoring disease conditions, estimating prognosis and guiding therapy in all kinds of leukemia. The simultaneous analysis of telomere length and cell surface antigen by multicolor flow-FISH may monitor abnormal clone or clonal evolution to predict recurrence more sensitively and specifically, and may provide a promising and widely applicable method for monitoring MRD in leukemia.

CD56 and CD11b antigen expressions in patients with acute monocytic leukemia and the clinical implications / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of cell surface differentiation antigen CD56 and CD11b antigen in acute monocytic leukemic (AML-M(5)) cells and their clinical significance.</p><p><b>METHODS</b>A total of 113 cases of de nove adult AML-M(5) were examined genetically and immunologically using G-banding technique, interphase fluorescence in situ hybridization (I-FISH) and flow cytometry immunophenotyping, and the results were analyzed in relation to their clinical data.</p><p><b>RESULTS</b>Of the 113 cases, the expression rates of CD56 and CD11b was 28.32% and 73.45%, respectively. The CD56(+) patients had high CD11b expression, and the expression levels of CD11b and CD56 were positively correlated (P<0.05). The incidence of karyotypic abnormalities was 48.57% (55 cases) in these patients, including 25 (22.12%) with 11q23 aberrations. Twenty-five cases were positive for MLL gene abnormalities as found by I-FISH analysis. Compared with the patients positive for both CD56 and CD11b, those negative for both CD56 and CD11b showed increased peripheral blood white blood cell (WBC) count and also increased blast and progenitor cells in the bone marrow (P<0.05); the former patients often had karyotypic abnormalities, commonly involving 11q23 aberrations (P<0.05), whereas the latter patients presented more likely with extramedullary infiltration and refractory leukemia (P<0.01) with lowered complete remission rate and shortened median survival time (P<0.01). CD56-positive patients were more likely to have karyotypic abnormalities and refractory leukemia than CD11b-postive patients (P<0.05), but the peripheral blood WBC counts, bone marrow blast and progenitor cells, extramedullary infiltration, complete remission rate or median survival time showed no significant differences between them (P>0.05).</p><p><b>CONCLUSION</b>AML-M(5) patients with CD56 positivity and high expression of CD11b often have aberrant karyotypes, commonly involving 11q23/MLL gene abnormality. These patients frequently develop extramedullary infiltration and refractory leukemia often with poor prognosis.</p>

Clinical characteristics and outcomes of 59 patients with acute lymphoblastic leukemia positive for BCR/ABL / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the clinical characteristics and outcomes of BCR/ABL-positive acute lymphoblastic leukemia (BCR/ABL360888725-ALL) and screen the prognostic factors for BCR/ABL360888725-ALL.</p><p><b>METHODS</b>From January 2001 to May 2008, 59 patients (median age of 32 years ranging from 3 to 69 years) with the diagnosis of BCR/ABL360888725-ALL by fluorescence in situ hybridization received induction chemotherapy with VDLP-/+Ara-C regimen. The patients who failed to respond to the chemotherapy received subsequent consolidation chemotherapy with imatinib (400-800 mg/day) (17 cases) or allogeneic hematopoietic stem cell transplantation (allo-HSCT) (16 cases).</p><p><b>RESULTS</b>Of the 59 patients, 32 (58.3%) achieved complete remission (CR) after the first induction cycle. In patients with peripheral white blood cell (WBC) count <30=10(9)/L, 30-99.9(9)/L and > or =100(9)/L, the CR rates were 75.0% (18/24), 56.3% (9/15) and 26.3% (5/19) (P=0.006), and the overall survival probability of 2 years ( OSs of 2-yrs) was 24.7%, 22.5% and 21.1%, respectively (P=0.180). According to the FAB classification, 56 cases were divided into L1, L2 and biphenotypic acute leukemia (BAL) subgroups, and their CR rates were 66.7% (6/9), 63.2% (24/38) and 22.2% (2/9) (P=0.029), with OSs of 2-yrs of 22.2%, 27.0% and 22.0%, respectively (P=0.623). In terms of immunophenotype grouping by EGIL, the patients with ALL, myeloid antigen-positive ALL and BAL had CR rates of 61.1% (11/18), 60.6% (20/33) and 12.5% (1/8) (P=0.039), and the OSs of 2-yrs of 22.7%, 21.0% and 18.8%, respectively (P=0.643). In 55 patients with known karyotype, the CR rates were 71.4%(5/7), 70.8% (17/24) and 37.5% (9/24) in normal, sole t(9;22) abnormality, t(9;22) with additional abnormalities groups (P=0.046), with the OSs of 2-yrs of 42.9%, 34.0% and 7.3%, respectively (P=0.000). The patients complicated by septicemia had significantly lower OSs of 2-yrs than those without septicemia (0% vs 38.8%, P=0.005). The OSs of 2-yrs were significantly higher in patients with consolidation chemotherapy with imatinib than those without (48.0% vs 11.2%, P=0.001), and allo-HSCT was associated with significantly higher OSs of 2-yrs than exclusive chemotherapy (54.2% and 8.5%, P=0.000).</p><p><b>CONCLUSION</b>BCR/ABL360888725-ALL with WBC> or =100 x 10(9)/L, presence of BAL diagnosed by FAB or FACM, t(9;22) with additional chromosome abnormalities all adversely affect the treatment results, and additional chromosome abnormalities and septicemia are associated with lower OSs of 2-yrs. Imatinib treatment and allo-HSCT can both improve the OSs of 2-yrs of the patients with BCR/ABL(+)-ALL.</p>

Effect of iodine excess on TRAIL and TRAIL-sR1 expression of thyroid in Balb/c and NOD mice / 中国地方病学杂志

Objective To investigate the influence of iodine excess on expression of TRAIl/TRAIL-sR1 in NOD and Balb/c mice and to study the effect of TRAIl/TRAIL-sR1 on the pathogenesis of experimental autoimmune thyroiditis(EAT). Methods Both Balb/c and NOD mice were divided randomly into control and iodine excess group by feeding with water containing no NaI or 0.05% Nal. The mice were sacrificed after 8 weeks. TRAIL and TRAIL-sR1 mRNA levels were detected by RT-PCR. The function, morphology and apoptosis of thyroids were also observed by ELISA and Tunnel stain. Results Treated by HI, enlarged follicles and flattened epithelium by accumulation of colloid were found in thyroids of both NOD and Balb/c mice. But significant lymphoid cell infiltration and local fibrosis were only found in thyroids of NOD HI group. The relative weight of thyroids of NOD mice in HI group[(104.8±14.5)mg/kg]was heavier than that of control group [(71.8±20.4)mg/kg]. The level of TT4 declined in HI group[(30.77±3.59)mmol/L]compared with control group[(36.43±2.66)mmol/L], meanwhile, the level of TSH was higher in HI group[(6.98±0.66)μg/L]than that in control group [(5.55±0.56)μg/L]. The difference being statistically significant(t=7.773,-9.526,-4.458, all P < 0.05). The relative weight of thyroids of Balb/c mice of HI group[(155.8±20.8)mg/kg]also heavier than that of control group [(105.1±22.0) mg/kg]. The level of TT4 droped in HI group [(19.75±3.32) mmoL/L]was higher than that in control group[(23.46±6.21)mmoL/L], the level of TSH in HI group[(4.14±1.71)μg/L]was higher than that in control group[(3.55±1.41)μg/L], the difference being statistically significant(t=7.554,-7.239,3.140, all P< 0.05). A great deal of apoptotie ceils observed in NOD (3.97±0.91) and Balb/c mice (1.05±0.45) by Tunnel stain were greater than control groups (0.21±0.15, 0.10±0.03), the difference being statistically significant in beth of the two species(t=-7.167,-17.772, both P < 0.05). The apoptosis index of thyroid follicular epithelium in NOD was obviously higher than Balb/c(t=-7.625, P<0.05). The level of TRAIL mRNA did not remarkably change in Balb/c between control group(0.000 59±0.000 39) and HI group(0.001 24±0.000 46, t=-1.940, P>0.05), but it increased apparently in NOD mice HI group(0.018 88±0.005 77) than that of control group(0.009 61± 0.00591, t=-2.71, P<0.05). The level of the expression of TRAIL-sR1 mRNA increased in HI groups of NOD (0.000 53±0.000 15) and Balb/c mice(0.000 42±0.000 09) than that in control groups of NOD(0.000 28± 0.000 05) and Balb/c mice (0.000 17±0.000 06) and the differences were statistically significant between the two species(t=3.050,3.990, all P<0.05). The differences of the expression of TRAIL and TRAIL-sR1 mRNA between the two species were significant(t=-3.37,-4.76, all P<0.05). Conclusions Iodine excess induces colloid goiter in beth species of mice and thyroiditis in NOD mice. The increase of TRAIL and TRAIL-sR1 influenced by iodine excess is one of the molecular bases of follicular epithelium apoptosis and inflammation in thyroids. Genetic factor is a key factor in the pathogenesis of thyroiditis.

Determination of eleutheroside B in antifatigue fraction of Acanthopanax senticosus by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue).</p><p><b>METHOD</b>The antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm.</p><p><b>RESULT</b>As1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6).</p><p><b>CONCLUSION</b>This HPLC method is simple,accurate and reliable.</p>

A new monoterpene glycoside from active fraction (DSS-A-N-30) of Danggui Shaoyao San / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of an active fraction (DSS-A-N-30) from Danggui Shaoyao San.</p><p><b>METHOD</b>DSS-A-N30 was prepared by macroporous resin chromatography, the compound was isolated by column chromatography on silica gel and RPC-18, the structure was elucidated by spectroscopic methods.</p><p><b>RESULT</b>A new monoterpene glycoside was isolated and identified from DSS-A-N-30.</p><p><b>CONCLUSION</b>The new monoterpene glycoside was identified as 4"-hydroxyl-albiflorin.</p>

Effects of graft compositions on hematopoietic reconstitution and graft-versus-host disease in related donor peripheral blood stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of graft composition on hematopoietic reconstitution and graft-versus-host disease (GVHD) in HLA-identical related donor peripheral blood stem cell transplantation (PBSCT) for hematological malignancies.</p><p><b>METHODS</b>The relationship between the number of graft composition and their hematopoietic reconstitution and GVHD in 107 patients with hematological malignancies undergoing HLA-identical related donor PBSCT was retrospectively analyzed.</p><p><b>RESULTS</b>None of the graft composition numbers had correlation with the time of neutrophil reconstitution. There was a negative correlation between the number of mononuclear cells (MNCs) or CD34+ cells and the time of platelet reconstitution (P < 0.05) with the absolute correlation coefficients below 0.4, and so did between the number of CD34+ or CD34+ CD38- cells and the development of acute GVHD (P < 0.05) and their absolute correlation coefficients. Each lymphocyte subset number had no correlation with acute GVHD. There was a positive correlation between the number of CD25+ CD4+, CD3+ or CD4+ CD3+ cells or CD4+ /CD8+ ratio and the development of chronic GVHD (P < 0.05). And the correlation coefficients all exceeded 0.4, more over, CD25+ CD4+ cells number reached up to 0.78. CD34+, CD34+ CD38- cells number had no correlation with chronic GVHD.</p><p><b>CONCLUSION</b>In HLA-identical related donor PBSCT, further increasing infused MNC, CD34+, CD34+ CD38- cells can no more accelerated hematopoietic reconstitution after these cells attained their threshold values, otherwise the incidence of cGVHD might increase due to the increase of the accompanied lymphocytes.</p>