Objective To study the double restriction fluorescent labeling (DRFL) method for fluorescent labeling of trace DNA samples and its effect in enhancing the pathogen detection sensitivity of microarray assays. Method SARS-CoV RNA samples were reversely transcribed and then further amplified with the restriction display (RD)-PCR and fluorescently labeled by conventional restriction labeling directly with Cy-universal primer and the novel double labeling with Cy-universal primer and CydNTP. The labeled samples were applied to the microarray with the viral probes, processed and analyzed. Results Compared with the conventional method, DRFL labeling resulted in 3. 5835 times higher fluorescent intensity of all the SARS probes on average, even though increased fluorescent intensities for different probes varied considerably. Conclusion Signal to noise ratio can be enhanced by the DRFL method which improves the sensitivity of microarray technology in trace pathogen detections.

Objective In the treatment of acute myeloid leukemia (AML-M2a), the first CCR (continuous complete remission) has been one of the most critical indicators to the prognosis of the patients. Using microarray approaches, gene expression profiles have been studied in patients with different CCR, in order to find out the genes relevant to the progresses of the AML. Methods Bone marrow mononuclear cells were collected and used as different experimental groups respectively. Group A composed of three AML patients with CCR<6 months, while group B composed of three AML patients with CCR>12 months. mRNAs were purified and labeled with Cy3 and Cy5 respectively, which were used to hybridize against the Agilent human 1B 60mer oligonucleotide microarrays. Results In the 20173 genes tested, 21 genes were found expressed differentially between these two groups. Of these differentially expressed genes, 10 genes were up-regulated while 11 genes were down-regulated in group A. Conclusion Through microarray studies, 21 genes including APP were found to be differentially expressed in AML patients whom were treated with standard chemotherapy. Theses genes can be early indicators for the diagnosis as well as prognosis of the refractory AML.

Objective To evaluate the therapeutic effects of allogeneic hematopoietic stem cell transplantat (allo-HSCT) for severe aplastic anemia (SAA). Methods Four patients of SAA underwent allo-HSCT at the bonemarrow transplant unit in our hospital from March 2003 to May 2009. Stem cell source was an HLA (human leukocyte antigen) matched related donor (MRD) in 3, HLA 1 (B) mismatched related donor in 1 patient A retrospective analysis was performed on interval from diagnosis to transplant,HSCT manners,conditioning regimens, hematopoiesis reconstitution, effectiveness and complication. Results The interval from diagnosis to transplant was 70 (19 - 180) days. Three patients (MRD) underwent BM + PBSCT, one was undergone BM + PBSC + CBSCT. Conditioning regimens of all patients were CY/ATG. Hematopoiesis reconstitution was achieved in 4 patients (100%). The median time of neutrophils which reached 0. 5 x 109/L and platelets reached 20 × 109/L were 14. 5 (9-28) and 16(9 -28) days. Two cases developed grade Ⅰ acute graft-versus-host diseaes (aGVHD), chronic local GVHD occurred in one patient. Four patients are alive with a median time of 40. 6(2 -63) months at the end of the following-up. Conclusions Allo-HSCT are an efficient and safe therapy for the patient with SAA,not only for patients with HLA matched related donor,but also for those only HLA mismatched related donor available.

Objective To explore the high-risk factors,clinical characteristics,therapy and prognosis of invasive fungal infection (IFI)in patients underwent allogeneic haemopoietic stem cell transplantation (AlloHSCT). Methods One hundred patients underwent Allo-HSCT at our department from March 2002 to July 2010 were analyzed retrospectively,among whom 26 patients had invasive fungal infection(IFI). Seven patients had pulmonary IFI before allo-HSCT, 14 patients had pulmonary IFI after allo-HSCT,3 patients had respiratory tract system IFI, and 2 patients had intestinal IFI. We observed the occurrence of Graft-versus-host disease (GVHD) ,cytomegalovirus( CMV )infection, Lymphocyte subsets and chronic basic diseases in patients with IFI. The twenty six cases were divided into two groups: experience therapy group with 12 cases and preemption therapy group with 14 cases. Results Among 26 patients with IFI,20 cases suffered from GVHD,6 cases had CMV infection,19 cases had low cellular immune function simultaneously. 1 case had diabetes,3 patients had pulmonary tuberculosis and 1 case had bronchiectasis as complications. In experience therapy groupe: 8 cases (67%)recovered completely but 1 case(8% )suffered from progressive infection. In preemption therapy groupe:3 cases ( 21% ) recovered completely but 5 cases ( 36% ) suffered from progressive infection. Conclusion Clinician should pay close attention to the patients with high-risk factors of IFI after allo-HSCT.

Objective To investigate the cytogenetic and immunological phenotypes of acute myeloid leukemia (AML) with t(8;21),and explore the risk stratification and risk-adapted treatments.Methods The chromosomal karyotype of bone marrow was detected and analyzed in 22 newly diagnosed patients with t(8;21) AML by direct culture and G banding technique.Patients were divided into two groups according to the chromosomal karyotypes.Clinical characteristics and immunological phenotypes were compared between patients with isolated t(8;21) and those with additional aberrations.A follow-up study with median time 30 months (4-68 months) was conducted to analyze prognostic factors.Results 13 cases (59.1％) were isolated t(8;21) AML,while 9 (40.9 ％) had additional aberrations.Loss of sex chromosome was found in 3 cases and complex variant translocation in 2.The 10q-,9q-,-18 and +10 were found in single cases.Overall survival of patients with additional aberrations was significantly poorer than those with isolated t (8;21) (P =0.0176).Analysis of prognostic factors showed that t(8;21) chromosomal karyotype,initial white blood cells at diagnosis,and treatment regimen (chemotherapy alone or plus hematopoietic stem cell transplantation) had effects on overall survival.Conclusion Patients with t (8;21) AML are frequently associated with additional chromosomal aberrations.The latter indicates a poorer outcome and can be one of the bases of risk stratification.Hematopoietic stem cell transplantation might help to improve the overall survival.

Objective: To evaluate the feasibility of full-mouth debridement ( subgingival scaling and root planning , SRP) by 2 times within 1 week and compare the clinical effects of different sequences of debridement-antibiotic usage in patients with severe chronic periodontitis ( CP ) .Methods: A double-blinded, placebo-controlled, randomized clinical trial was conducted in 30 severe CP patients (14 males and 16 females, 40.5 ±8.4 years old on average from 35 to 60 ) receiving 3 different sequences of debridement-antibiotictherapy:Group A, antibiotic usage (metronidazole, MTZ, 0.2 g, tid, 7 d;amo-xicillin, AMX 0.5 g, tid, 7 d) was started together with SRP ( completed by 2 times in 7 d);Group B, antibiotic usage (MTZ 0.2 g, tid, 7 d;AMX 0.5 g, tid, 7 d) was started 1 d after SRP(completed by 2 times in 7 d);Group C, SRP alone[probing depth (PD), bleeding index (BI) and tooth mobility] was examined .The average full-mouth probing depth , the average full-mouth proximal probing depth ( pPD) , the percentage of sites with PD >5 mm ( PD>5 mm%) , the percentage of sites with proximal PD>5 mm ( pPD>5 mm%) , the average bleeding index ( BI) and the percentage of sites with bleeding on probing ( BOP%) were calculated .Clinical examinations were performed at baseline and 2 months post therapy .Results:(1) Compared with baseline conditions , all the subjects showed clinical improve-ments in all the parameters evaluated 2 months post therapy , P<0 .05 .( 2 ) Significant difference were observed in the average PD changes between Group A [(2.15 ±0.42) mm], Group B [(1.76 ±0.29) mm] and Group C [(1.57 ±0.33) mm], P<0.05.No significant difference was observed in the aver-age PD changes between Group B and Group C , P=0.354.Significant differences were observed in the average pPD changes between Group A [(2.45 ±0.43)mm] and Group C[(1.90 ±0.48) mm], P<0.05.No significant difference was observed in BI and BOP% changes between Group A ,Group B and Group C.Conclusion: For patients with severe chronic periodontitis , it is safe and feasible to receive full-mouth SRP by 2 times within 1 week.The short-term ( 2 months ) advantages in PD changes are observed in patients receiving SRP and antibiotic usage at the same time comparing with patients using antibiotics after SRP or SRP alone .

Objective To explore the relationship of cell proliferation of acute leukemia stimulated by granulocyte colony-stimulating factor(G-CSF) and the expression of granulocyte colony stimulating factors receptor(G-CSFR).Methods Thirty cases of initial and refractory-relapse acute myeloid leukaemia(AML) patients,20 cases of acute lymphoblastic leukemia(ALL) and 20 normal controls were involved in the study.The 5ml bone marrow was taken from each patient before chemotherapy and the marrow mononuclear cells(MNC) were cultured with 5,10,15,20 and 25 ng/ml G-CSF respectively.After 24h,the DNA diploid and the expressions of G-CSFR and CD34 were detected by flow cytometry(FCM).Results The DNA diploid of MNC from AML was increased with the elevated concentration of G-CSF after 24h,and that of the ALL and normal control did not change significantly.The expression rates of G-CSFR were(68.59?13.99)%,(1.90?0.93)% and(70.5?10.8)% in AML,ALL and the normal control respectively.The expression rates of CD34 were(45.15?4.22)%,(46.75?3.15)% and(3.15?0.22)% in AML,ALL and normal control respectively.The expression of G-CSFR in AML was significantly different from that of ALL(P0.05).The expression of G-CSFR in ALL was significantly different from that of the normal control(P

ObjectiveTo explore the clinical significance of fms-like tyrosine kinase3(FLT3) expression in evaluation prognosis of acute myeloid leukemia(AML) prognosis.Methods50 patients with AML were selected.AML patients with normal karyotype were 20 cases,the abnormal karyotype were 30 cases.3ml bone marrow before themotherapy was aspirated respectively,and the FLT3 gene expression in leukemia cells was detected with polyenzyme chain react(PCR).ResultsThe FLT3 expression rate in AML patients with normal karyotype was 5.0％,and was 26.7％ in AML patients with abnormal karyotype,33.3％ in AML patients with refractory-relapse,and 4.5％ in AML patients with continue remission.The FLT3 expression rate was related with high leukemia cells percentage in bone marrow and high blood cells count in peripheral blood,and was not related with Franch America British(FAB) classification.The free-disease survival(FDS) and overall survived(OS) was shorter in FLT3 expression AML patients than that in no FLT3 expression AML patients.There was a statistical significance between the former and the latter( x2 =4.17,P ＜0.05 ).Conclusion FLT3 was a kind of worse factor in AML patients prognosis,and could guide clinical individual treatment in AML.

To investigate the effects of Ligustrazine on histogenesis of bone marrow in the early phase of hematopoietic reconstruction in bone marrow transplantation (BMT) mice. The syngeneic BMT mice model was established. The syngeneic BMT mice were orally given 2 mg Ligustrazine twice a day. 1, 3, 5, 7, 10, 15 and 21 day(s) after BMT, peripheral blood granulocytes and bone marrow nucleated cells (BMNC) were counted and the diameter of central vein and the area of micro-vessel in femur were measured. The effect of Ligustrazine on hematopoietic stem cells was observed by colony forming unit of spleen (CFU-S). The effect of Ligustrazine on hemopoietic progenitors was studied by observing the number of progenitors of Granulocytes/Macrophage on day 10 and day 20 after BMT. In Ligustrazine-treated group, the diameter of center veins and the area of micro-vessel of femur were all significantly less than the control group 7, 10, 15, 21 days after BMT (P < 0.01). In addition, Ligustrazine significantly increased the number of CFU-S on day 10 and the number of CFU-GM on day 10, 20 after BMT. These results indicate that Ligustrazine can accelerate the histogenesis of hemopoietic bone marrow, which may be one mechanism by which Ligustrazine promotes hematopoietic reconstitution after BMT.
*Bone Marrow Transplantation ; Hematopoiesis/*drug effects ; Hematopoietic Stem Cells/*drug effects ; Mice, Inbred BALB C ; Pyrazines/*pharmacology ; Time Factors

Objective:To observe the effect of electromyographic biofeedback on the wrist dirsiflexion function of the patients with cerebral infarction at different Brunnstrom stages, and to clarify the treatment of electromyographic biofeedback,and to provide basis for its clinical application.Methods:A total of 100 cerebral infarction patients were selected.Among them 54 BrunnstromⅠ-Ⅱ patients were randomly divided into treatment group (n= 32)and control group (n = 22),and another 46 Brunnstrom Ⅲ patients were randomly divided into treatment group (n=23)and control group (n=23).The patients in four groups were treated with the same routine stroke rehabilitation therapy while the patients in treatment groups still received the electromyographic biofeedback therapy additionally.The maximum electromyographic contraction of muscle,active range of movement (AROM) and Fugl-Meyers Assessment (FMA)of the extension of wrist joint were evaluated before treatment and 4 and 8 weeks after treatment,respectively.Results:The maximum electromyographic contraction values of muscle of the patients in BrunnstromⅠ-Ⅱ treatment group and control group were significantly improved 8 weeks after treatment (P <0.05),and the value in treatment group was higher than that in control group (P <0.05).The maximum electromyographic contraction value of muscle in Brunnstrom Ⅲ treatment group began to improve 4 weeks after treatment compared with before treatment (P < 0.05) and it was significantly higher than that in control group (P <0.05).The maximum electromyographic contraction value of muscle in Brunnstrom Ⅲ control group began to improve 8 weeks after treatment (P <0.05).The AROM in Brunnstrom Ⅰ-Ⅱ treatment group began to improve 8 weeks after treatment (P <0.05)and it was significantly higher than that in control group (P <0.05)while the AROM in control group had no significant change (P >0.05).The AROM in Brunnstrom Ⅲ treatment group and control group were significantly improved 4 weeks after treatment (P < 0.05 or P < 0.01 ), and the value in treatment group was significantly higher than that in control group (P < 0.05).The FMA in BrunnstromⅠ-Ⅱtreatment group and control group were significantly improved 8 weeks after treatment (P <0.05),while the value in treatment group was higher than that in control group (P <0.05);the FMA in Brunnstrom Ⅲ treatment group began to improve 4 weeks after treatment (P < 0.05)and it was significantly higher than that in control group (P <0.05). The FMA in control group began to improve 8 weeks after treatment (P <0.05). Conclusion:Electromyographic biofeedback can increase the strength and improve the body function of the patients with cerebral infaction.