Objective To study the mechanism of the extract of three kinds of Chinese herbs in treating psoriasis.Methods Three kinds of Chinese herbs(coptis root,patience dock root and atratylodes rhizome) were extracted.With the culture of human transformed keratinocyte cell line colo-16 cells as a model,the single and compound effects on the growth of KC were studied before and after stimulated by TNF-?.Results The single and compound extract of three Chinese herbs could restrain the proliferation of KC before and after stimulated by TNF-?.The effect was more remarkable than the compared methotrexate and cyclosporin A(P

Objective To establish and evaluate the new method of 8-color flow cytometry (8c-FCM) with two tube detecting bone marrow minimal residual disease (MRD) in B lineage acute lymphoblastic leukemia (B-ALL).MethodsThe MRD cells were analyzed by using two combinations of 8c-FCM antibody panels,gating with CD19/Side scatter(SSC),CD45/CD10 and CD34(or cTdT).Nalm-6 cell of B-ALL was mixed into normal marrow cells,with proportion of 10.00％,1.00％,0.10％,0.01％and 0.005％,and recovery test and reproducibility test were carried with 8c-FCM established to value its accuracy and precision of.Fluorescence intensity was detected on different time points after marked Nalm-6 by antibodies to evaluate the fluorescent stability of the antibodies.The immunophenotyping was analyzed in 39 bone marrow specimens,including 9 cases of normal control,9 cases of B-ALL primary or recurrent,21 cases of complete remission (CR) after chemotherapy or bone marrow transplantation ( BMT),to evaluate the detection of normal B lymphocyte lineage,leukemia cells of B-ALL,MRD cells of B-ALL using the 8c-FCM and compare it with 4c-FCM in being.ResultsThe method of two tube 8c-FCM with main antibodies of CD19,CD45 and CD10 to detect MRD of B-ALL was founded; CV ＜ 2.5％,average recovery rate was 95.81％,when the actual percent of Nalm-6 mixed into normal bone marrow≥0.10％,and the percent of Nalm-6 detected and actual was linear dependent ( r =0.99,P ＜ 0.05 ) ranged from 10.00％ - 0.01 ％,when 106 cells were acquired ; the sensitivity of the method established could reach 0.01％.The fluorescent intensity decreased along with the time after Nalm-6 cell marked,but less than 10％ in 24 hours.Using the antibody combinations and analyze strategy,the immunophenotye of B lymphocyte in normal bone marrow presented four sequential stages:Stage Ⅰ CD45low/CD10stro/CD20 -/CD38stro/CD34 + or cTdT +,Stage ⅡCD45 +/CD10 + / CD20 -/CD38stro/CD34+ or cTdT +,Stage Ⅲ CD45 +/CD10 +/CD20 -/CD38stro/CD34 -or cTdT-,Stage Ⅳ CD45stro/CD10 -/CD20 +/CD38low/CD34or cTdT.Antigen expressions of leukeamic cells of B-ALL primary or recurrent were different compared with control team; there were 5 cases with MRD positive in CR team,and the main antigen expression was consistent with the results from 4c-FCM.The range of the percent of MRD cells was 0.02％ - 5.42％ of the 5 cases of MRD positive.ConclusionsThe new method of two tube 8c-FCM established shows good reproducibility and high accuracy,and can identify normal B lymphocyte populations in bone marrow and regenerated B-precursors in CR cases with MRD cells;compared with 4c-FCM,the new method of two tube 8c-FCM with the fewer specimen is faster and efficient to diagnose MRD of B-ALL.

Objective To explore the values of potential clinical application ofsingle tube/ten colorsflow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency ofsingle tube/10 colorsflow cytometry was calculated.The correlations between standard microscopic cytology,single tube/10 colorsflow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts ofsingle tube/10 colorswere compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the single tube/10 colorsflow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity ofsingle tube/10 colorsflow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts bysingle tube/10 colOrsflow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions Thesingle tube/10 colorsflow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.

Bronchial Diseases ; diagnosis ; Bronchial Neoplasms ; diagnosis ; Humans ; Male ; Middle Aged ; Mucormycosis ; diagnosis

Objective To assess the efficacy and safety of monoclonal antibody rituximab combined with cyclophosphamide (CTX) in the treatment of refractory and recurrent autoimmune hemolytic anemia.Methods Seven cases with refractory and recurrent autoimmune hemolytic anemia ( including 1 case of Evans syndrome) were recruited during January,2007 to December,2010.Treatment regimens were as follows:rituximab:375 mg/m2,1 time/week,2-6 courses; CTX:1 g,1/10 d,2-7 courses; combined with intravenous immunoglobulin (IVIG) 5 g,1 time/week,given 1 day after rituximab administration.The efficacy and safety of this regimen were assessed during follow-up.Results All the patients showed good responses (7/7).Six patients achieved complete remission (6/7) and one achieved partial remission ( 1/7 ).Average follow-up time for the patients was 27 months.All patients remained in remission during the 12-month follow-up visits.Two patients showed elevated indirect bilirubin and increased reticulocyte counts within 24 months.One patient achieved complete remission after additional rituximab therapy,and another patient remained partial remission after cyclosporine therapy.At the time of 36-month follow-up visit,the patient relapsed and was retreated with 3 courses of rituximab combined with CTX and eventually achieved partial remission.All patients tolerated the treatment well with few mild side effects.Conclusions Rituximab combined with CTX is effective and relatively safe in patients with refractory and recurrent autoimmune hemolytic anemia.Additional treatment to relapse patients about 12-24 months after drug withdrawal continues to be effective.

This study was aimed to investigate the clinical features of patients with immune thrombocytopenia (ITP). The clinical data, laboratory results and therapeutic effects of 125 inpatients with ITP from January 2005 to May 2010 were retrospectively analyzed. The results showed that median age of 125 inpatients was 41, and the sex ratio of male to female was 1:1.78. The average pre-treatment platelet count was (28.59 ± 23.05) × 10(9)/L. 69.1% patients (67/97) suffered from rheumatoid or immune abnormalities. The carrier rate of hepatitis B virus and parvovirus B19 were 37.3% (22/59) and 44.8% (26/58) respectively. CD3(+)T cells in chronic patients was (62.7% ± 14.11%), while that in acute patients was (55.44% ± 14.31%). The rate of CD19(+)B cells in chronic patients was (12.83% ± 6.96%), that was (19.47% ± 6.93%) in acute patients. 91 patients were subjected to the examination for bone marrow mononuclear cell membrane antibody, with 54.9% (50/91) cases negative and with 45.1% (41/91) cases positive. 63.0% (17/27) acute patients were positive, while that was 37.5% (24/64) in chronic patients. Therapy of hormone combined with cyclosporine showed efficiency of 81.7% (49/60), the response of some refractory patients to RCP chemotherapy or azathiopurine cytotoxic drugs showed efficiency of 57.14% (4/7) and 40.0% (2/5) respectively. It is concluded that patients with ITP always accompany with virus infection or rheumatism abnormalities. Acute patients always accompany with hyperfunctioning of humoral immune, while the chronic ones were with hyperfunctioning of the cellular immune. Patients with positive bone marrow mononuclear cell membrane antibody are more likely to be acute. In order to obtain a better effect, the treatment strategies should be based on different pathogenesis to choose different drugs or different combinations of medicine.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibody Formation ; Child ; Child, Preschool ; Female ; Humans ; Immunity, Cellular ; Male ; Middle Aged ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; immunology ; virology ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the impact of 36-hour sleep deprivation (SD) on the brain electrophysiological indicators of visuo-motor coupling in young soldiers.

METHODSDuring the 36-hour SD, 10 healthy young soldiers were tested on visuospatial rotation tasks by event-related potentials system before and after SD. The incubation period and amplitude of P500 as well as their error number and reaction time were measured.

RESULTSCompared with subjects in SD 0-hour,subjects in SD 36-hour had significantly increased error rate [(9.7 ± 3.9)% vs. (18.3 ± 4.5)%, P<0.05] and significantly increased reaction time [(632.5 ± 53.6) ms vs. (693.6 ± 65.7) ms, P < 0.05]. Subjects in SD 36-hour showed significantly reduced amplitudes than those in SD 0-hour [(8.7 ± 2.3) ΜV vs. (5.2 ± 1.6) ΜV, P < 0.05]. Additionally, subjects in SD 36-hour showed significantly increased P500 latencies than did those in SD 0-hour [(489.6 ± 42.6) ms vs .(530.2 ± 51.9) ms, P < 0.05]. Compared with subjects in SD 0-hour, the deficit was an absence of a mental rotation function SD 36-hour in subjects.

CONCLUSIONSThe 36-hour SD in young soldiers can harm the processing mechanism of visuo-motor coupling in a certain extent. SD can affect the fixed position ability of visual space cognition in young soldiers.

Adolescent ; Adult ; Evoked Potentials ; Feedback, Sensory ; Female ; Humans ; Male ; Military Personnel ; Reaction Time ; Sleep Deprivation ; physiopathology ; Young Adult

Amino Acid Sequence ; Base Sequence ; Genes, Bacterial ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Staphylococcus epidermidis ; enzymology ; genetics ; beta-Lactamases ; chemistry ; genetics

Objective To study the effect of iron overload on the bone marrow hematopoietic function on immuo-related pancytopenia(IRP)patients by hematopoietic progenitor cell(HPC)culture of bone marrow(BM).Methods BM liquid 4～5 mL was taken from 46 IRP patients of General Hospital Tianjin Medical University from July 2009 to February 2010 to detect colony-forming-unit of erythrocyte,blast-forming-unit of erythrocyte and colony-forming-unit of granulocyte-monocyteby HPC culture of BM.And according to the serum ferritin(SF)level,these patients were classified into 2 groups to compare HPC proliferation,blood cell counts,BM proliferation,blood transfusion and treatment effects.Results The mean values of 3 different colonies of IRP patients with high SF level [(43.33?17.74),(1.50?2.2),(11.06?5.83)/105BMMNC] were significantly lower than those of the patients with normal SF level [(77.43?40.64),(9.57?7.99),(21.25?11.41)/105BMMNC](P

BACKGROUNDHIWI is a member of PIWI gene family and its expression is found in various tumors, indicating that it may play a pivotal role in tumor development. This study was designated to examine HIWI protein expression profile in several cancer cell lines and its prognostic value for patients with colorectal cancer.

METHODSTotally 270 patients who underwent surgical resection of primary colorectal cancer between January 1999 and December 2002 with a median follow-up time of 33 months were registered in the study. Formalin-fixed and paraffin-embedded specimens from these patients and 236 matched adjacent non-cancerous normal colorectal tissues were collected. Anti-HIWI monoclonal antibodies were generated and used for evaluating HIWI protein expression. χ(2) tests were conducted to determine the association between HIWI expression and the other variables. Survival curves were estimated using the Kaplan-Meier method and compared by the log-rank test. Multivariate analysis was performed by using the Cox regression model.

RESULTSBy generating antibodies specific for HIWI, we examined HIWI protein expression in several cancer cell lines and demonstrated positive expression of HIWI in 69 out of 270 (25.6%) colorectal cancer tissues; 15 of 236 (6.4%) matched adjacent non-cancerous tissues were also positive for HIWI. Patients with positive HIWI expression in adjacent non-cancerous tissue had statistically lower overall survival (OS) and disease free survival (DFS) compared with negative patients (OS: 10.4% vs. 55.5%, P = 0.009; DFS: 10.4% vs. 55.1%, P = 0.015). For early stage group (stages I and II), patients with positive HIWI expression had significantly lower OS and DFS (OS: 57.4% vs. 79.5%, P = 0.014; DFS: 56.7% vs. 80.5%, P = 0.010). In lymph node negative group, patients with positive HIWI expression had statistically lower OS and DFS (OS: 53.0% vs. 73.5%, P = 0.037; DFS: 52.2% vs. 74.6%, P = 0.025). Multivariate analysis revealed that HIWI over-expression was a significant prognostic factor for OS (95%CI: 1.132 - 2.479, P = 0.010).

CONCLUSIONHIWI could be a potential prognostic biomarker for the patients with colorectal cancer, especially for those at early stages or without lymph node metastasis.

Argonaute Proteins ; metabolism ; Blotting, Western ; Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Prognosis