Objective To evaluate the utility of flow cytometry (FCM) in diagnosis and subclassification of non-Hodgkin lymphoma (NHL). Methods The samples of lymph nodes biopsy from 59 cases clinically suspected of NHL were detected by flow cytometry; and clonal lymphocytes and their immunophenotypes were identified analyzed. The concordance between the results of flow cytometry and histopathology was analyzed. Results Among the 59 cases, flow cytometry was able to identify aberrant clonal lymphocytes in 24 of 28 NHL cases identified by histopathology, the neoplastic lymphocytes ranged from 4.28 % to 89.10 %; 23 cases were diagnosed as B-NHL and 1 case was diagnosed as T-NHL. Compared with histopathology, the accuracy of FCM was 85.71% in diagnosis of NHL. The specificity and sensitivity of FCM was 100 % and 92% in diagnosis of B-NHL. The accuracy of flow cytometry immunophenotyping in classification of 24 cases of NHL was consistent with that of histopathology. Conclusion Flow cytometry could be an ancillary technique in diagnosis of NHL by identifying aberrant clonal lymphocytes, and enable identification of B-NHL subtype.

Objective To explore the clinical efficacy and value of azo steen,montelukast sodium combined with levocetirizine in the treatment of chronic urticaria. Methods 150 cases with chronic urticaria admitted in department of dermatology of Hubei Zhongshan Hospital were divided into A,B and C group,each group had 50 cases. Patients in group A were received montelukast sodium and levocetirizine treatment,which in group B were received montelukast sodium and azo steen treatment,group C were received above three kinds of drug treatment. Total effective rate,adverse reaction and safety in three groups were observed and compared. Results Total effective rate of group C was significantly higher than A and B group(P<0.05), and the safety of group C were better than A and B group(P<0.05 ),too. The occurrence of adverse reaction in group C was significantly less than that in group A and B (P <0.05 ).Conclusion Azo steen,montelukast sodium combined with levocetirizine has good efficacy in treatment of chronic urticaria. It can relieve itching wheals and other symptoms in a short time,with less adverse reaction.

OBJECTIVETo compare the difference in the clinical efficacy on cervical spondylosis of vertebral artery type (CSA) treated with thermosensitive moxibustion at different dosages.

METHODSSixty cases of CSA were randomized into a saturated moxa dosage group and a regular moxa dosage group, 30 cases in each one. The thermosensitive moxibustion was adopted in the two groups. The mild suspended moxibustion was applied at two acupoints with the strongest thermosensitization. In the saturated moxa dosage group, the moxibustion time was determined by the disappearance of thermosensitization. In the regular moxa dosage group, 15 min was required on each acupoint. The treatment was given twice a day for first 4 days in the two groups. Since the 5th day, the treatment was given once a day, continuously for 10 times, and totally 14 days were required. The score of symptoms and function and clinical efficacy were compared between the two groups before and after treatment as well as 6-month follow-up after treatment.

RESULTSThe curative and effective rate was 56.7% (17/30) after treatment and 60.0% (18/30) in 6-month follow-up after treatment in the saturated moxa dosage group, which were superior to 26.7% (8/30) and 30.0% (9/30) in the regular moxa dosage group respectively (P < 0.01, P < 0.05). The scores of clinical symptoms and function after treatment and in follow-up were improved apparently as compared with those before treatment in both groups (all P < 0.01). The scores of clinical symptoms and function after treatment and in follow-up in the saturated moxa dosage group were increased much more apparently than those in the regular moxa dosage group (after treatment: 22.32 +/- 4.64 vs 17.43 +/- 3.21; in follow-up: 23.01 +/- 4.76 vs 18.32 +/- 2.13, both P < 0.01).

CONCLUSIONThe thermosensitization moxibustion of saturated dosage achieves the superior short-term and long-term efficacies in the treatment of CSA as compared with the regular moxibustion dosage.

Acupuncture Points ; Adult ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; instrumentation ; Spondylosis ; physiopathology ; therapy ; Vertebral Artery ; physiopathology

The study was aimed to investigate the application value of interphase fluorescence in situ hybridization (FISH) on cell smears in hematological diseases. Both interphase FISH on peripheral blood smears and bone marrow smears treated by methanol/acetic acid, and routine interphase FISH of bone marrow cells dropped on slides were done at the same time, in order to detect Ph chromosome by BCR/ABL dual color, dual fusion probe in 20 patients with chronic myelogenous leukemia or acute lymphoblastic leukemia which had been proven to display Ph chromosome positive. The results indicated that as compared with routine interphase FISH, the interphase FISH on cell smears could also offer reliable result. It is concluded that interphase FISH on cell smears is a kind of reliable and time-saving technique, which is also suitable for retrospective research and worthy to further apply in clinic.
Adult ; Aged ; Cytogenetic Analysis ; methods ; Female ; Hematologic Diseases ; diagnosis ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Interphase ; genetics ; Male ; Middle Aged ; Young Adult

The objective of study was to evaluate the clinical values of multiparameter flow cytometry (MPFC) and cytomorphology of bone marrow aspiration(BMA) in detecting bone marrow involvement in patients with B cell Non-Hodgkin's lymphoma (B-NHL). 96 bone marrow samples from the patients with B-NHL were measured by MPFC using CD45/SSC and CD20/SSC gating strategy combined with anti-kappa and anti-lamda monoclonal antibodies, and then compared with results acquired by cytomorphologic analysis of BMA. The results showed that the bone marrow involvement was confirmed by MPFC in 38 cases (39.6%), while it was detected by cytomorphologic analysis of BMA only in 12 cases (12.5%). There was a significant difference between the two methods (p<0.05). 12 positive cases detected by cytomorphologic analysis of BMA were also positive by MPFC. There was no difference of 3-year overall survival rate between negative and positive cases detected by MPFC, but their 4-year overall survival rate was 73.18+/-6.65% and 44.13%+/-19.55% respectively (p<0.05). It is concluded that the MPFC is a more sensitive method for detecting bone marrow involvement in patients with B-NHL than cytomorphologic analysis of BMA. The 4-year overall survival rate of the patients without bone marrow involvement was significant higher than those of patients with bone marrow involvement. Bone marrow involvement in B-NHL detected by MPFC can be useful for clinical evaluation and prognosis prediction.
Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; pathology ; Female ; Flow Cytometry ; methods ; Humans ; Lymphoma, B-Cell ; pathology ; Lymphoma, Non-Hodgkin ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Survival Rate ; Young Adult

Objective To investigate the effect of laser artificial shrinkage(LAS)on pregnancy outcome in vitrification of human expanded blastocysts.Methods We selected 3859 frozen-thawed blastocyst-stage embryo transfers from January 2014 to December 2015.The transfers were divided into LAS group(n=3 176)and non-LAS group(n=683),which were then subdivided into ＜36 y subgroup and ≥36 y subgroup according to their age.Main outcomes measures were thawing rate,implantation rate and clinical pregnancy rate.Results Thawing rate, clinical pregnancy rate and implantation rate were 97.32%(5 453/5 603),66.81%(2 118/3 170),and 53.55%(2 912/5 438)in LAS group.In non-shrink group,they were 95.13%(1 173/1 233),62.70%(427/681),and 49.74%(582/1 170),which did not significantly differ from those in the former group(P＜0.05).Further analysis of the subgroups showed that thawing rate was significantly higher in LAS group than in non-shrink group of patients＜36 y(97.27% vs.95.33%;P＜0.05).Thawing rate and biochemical pregnancy rate were significantly higher in LAS group than in non-shrink group in patients ≥36 y(97.75% vs.93.66%;65.45% vs.50.65%,P＜0.05). Cancellation rate was not significantly different between the two groups(0.19% vs.0.29%, P ＞ 0.05). Conclusion LAS technique can increase thawing rate,clinical pregnancy rate and implantation rate before cryopreservation of blastocysts.

OBJECTIVETo suppress COL1A1 and COL3A1 gene expressions in human skin fibroblasts (HSFs) by means of RNA interference (RNAi).

METHODSThree small interfering RNA (siRNA) expression cassette (SEC) sequences were designed for each of the COL1A1 and COL3A1 gene sequences available in GenBank. The synthesized SECs capable of effective gene suppression were transfected into cultured HSFs, either after cloning into the expression vector or mediated by Lipofectamine 2000, and the suppression of the target genes at both mRNA and protein levels was determined by quantitative fluorescence RT-PCR and Western blotting, respectively.

RESULTSTransfection of the SECs into HSFs resulted in specific depression of COL1A1 and COL3A1 expressions (down to 5.00% and 6.48%, respectively). The expression vector-mediated RNAi established a HSF cell line with persistent gene knockdown for over 30 days (to 25.21% and 22.12%, respectively).

CONCLUSIONCOL1A1 and COL3A1 gene expressions can be specifically and efficiently inhibited in HSFs by either liposome- or vector-mediated SEC transfection.

Blotting, Western ; Cells, Cultured ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Fibroblasts ; cytology ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Skin ; cytology ; Transfection ; methods

OBJECTIVETo screen the severe acute respiratory syndromes (SARS) mimotopes with random phage peptide library and to investigate their immunogenicity.

METHODSUsing SARS sera as selective molecule, a 12 mer phage peptide library was biopanned and positive clones containing the mimic epitopes were selected. The immuno-characteriation of the epitopes were then investigated.

RESULTS2 positive clones that having specific affinity to SARS sera were obtained. The DNA sequencing data showed no homology between the sequences of the deduced amino acid of the two mimic antigen peptides and the sequence of SARS.

CONCLUSIONSARS mimotopes were obtained by phage peptide library screening. This method might provide a new approach for SARS therapy and vaccine development.

Animals ; Antigens, Viral ; Base Sequence ; Biomimetic Materials ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; genetics ; immunology ; Humans ; Peptide Library ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; immunology

BACKGROUNDGlucocorticoid is speculated to be able to have Aspergillus fumigatus (A. fumigatus) being more susceptible to reactive oxygen species (ROS) by inhibiting Afyap1, the transcription factor activating protein-1 (AP-1) homologue in A. fumigatus, which may provide a clue to expand the clinical use of glucocorticoid in patients with fungal infections. In this study, we used dexamethasone to determine the direct effect on oxidative killing susceptibility of A. fumigatus in vitro, as well as the expression level of Afyap1 gene and its target genes (catalase and superoxide dismutase (SOD) genes).

METHODSA. fumigatus spores were treated with different concentrations (0, 0.02, 0.2 mg/ml) of glucocorticoids and assigned to four groups (A: 0.5 hour, B: 2 hours, C: 7 hours, D: 16 hours) according to the time of treatment. The H2O2 oxidative killing assay was done, using the standard method-spot test, in each group of A. fumigatus. We measured the oxidative killing susceptibility as well as the expression level of the gene Afyap1, CATA, SOD1 and SOD2 in A. fumigatus at each group. The antifungal susceptibility to itraconazole and amphotericin B in each group of A. fumigatus was also measured with M38-A2 method.

RESULTSThe oxidative killing susceptibility of A. fumigatus was increased, consistent with the reduction of Afyap1, CATA, SOD1 and SOD2 gene expression level after being treated with dexamethasone for 0.5 hours. However, these observations were disappeared along with being treated for longer time. The antifungal susceptibility to itraconazole and amphotericin B in the A. fumigatus strains treated with dexamethasone indicated no change, compared with those without dexamethasone treatment.

CONCLUSIONDexamethasone can have A. fumigatus being more susceptible to ROS when treated for shorter period (0.5 to 2 hours) via the reduction of Afyap1 gene expression as well as the down-stream enzyme-coding gene expression.

Aspergillus fumigatus ; drug effects ; genetics ; metabolism ; Dexamethasone ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Hydrogen Peroxide ; pharmacology

OBJECTIVETo screen anti-c-Met Fab from a phage antibody library and identify its binding activity.

METHODSThe expression of c-Met of HCC lines was identified by Western blot and immunofluorescence. Antibodies against c-Met were screened with immobilized antigen. After five rounds of panning, 30 randomly selected clones were identified by phage ELISA to select specific clones with high affinity. The positive clones were selected for Fab soluble expression in TOP10F and the binding activities were analysed in HCC lines.

RESULTSc-Met expressed in HCC membrane was confirmed by Western blot and immunofluorescence. A Fab fragment named AM2-26 with fine activity to c-Met was selected. AM2-26 binding specificity was confirmed by IP, FACS and immunofluorescence.

CONCLUSIONThe anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.

Antibodies ; immunology ; isolation & purification ; Cell Line, Tumor ; Cloning, Molecular ; Gene Library ; Humans ; Immunoglobulin Fab Fragments ; immunology ; Immunoglobulin Variable Region ; immunology ; Peptide Library ; Proto-Oncogene Proteins c-met ; immunology ; Recombinant Fusion Proteins ; immunology