OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo investigate the expression of HPA and HIF-1alpha in human esophageal squamous cell carcinoma and their relationship with cancer development, invasion and metastasis.

METHODSThe expression of HPA mRNA and HIF-1alpha protein in 23 mucosa specimens of incisal margin, 26 para-tumorous dysplastic tissues and 70 esophageal squamous cell carcinoma specimens were detected by in situ hybridization assay and immunohistochemical staining, respectively.

RESULTSThe positive expression of HPA mRNA and HIF-1alpha protein were significantly increased as the epithelial cells progressed into carcinoma (P < 0.05). The expression of HPA mRNA and HIF-1alpha protein in the esophageal squamous cell carcinoma were significantly correlated with the invasion depth, lymph node metastasis and clinical staging (P < 0.05), while it was not correlated with the differentiation of tumors (P > 0.05). There was a close correlation between the expression of HPA mRNA and HIF-1alpha protein in the carcinoma tissues (P < 0.01).

CONCLUSIONThe high expression of HPA mRNA and HIF-1alpha protein is correlated with carcinogenesis, progression, invasion and metastasis of esophageal squamous cell carcinoma. There may be a positive cooperation between two of them in the carcinogenesis and development of esophageal carcinoma. The determination of HPA mRNA and HIF-1 alpha will be useful in predicting tumor biological behavior.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Esophageal Neoplasms ; metabolism ; pathology ; Esophagus ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Glucuronidase ; genetics ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Lymphatic Metastasis ; Male ; Middle Aged ; Mucous Membrane ; metabolism ; Neoplasm Invasiveness ; Neoplasm Staging ; RNA, Messenger ; metabolism

Objective To understand epidemiological characteristics of chronic obstructive pulmonary disease(COPD)in people aged over 40 years in a rural area of Tianjin.Methods Using cluster sampling,1 508 subjects over 40 years old at five villages in Xinkaikou Township,Baodi District,Tianjin were investigated with respiratory questionnaire,lung function test and physical examination.Confirmed patients with COPD were examined by chest roentgenography and electrocardiography.Results One hundred and forty-two subjects in that area suffered from COPD,with prevalence of 9.4%,24 of them (16.9%)were diagnosed as cor pulmonale.Prevalence of COPD increased with age,higher in men (13.5%)than that in women(6.2%),higher in smokers(12.2%)than that in non-smokers(7.2%), higher in those with family history(21.4%)than that in those without it(8.45%),and higher in those with coughing history during their childhood(75.0%)than that in those without it(9.2%),all with a P-value of less than 0.01.Univariate analysis showed that out-door air pollution,cooking,time length of burning firewood during cooking,smoking,coughing history during childhood,gender,age,family history all were predisposing factors for COPD.Multivariate analysis with logistic regression model showed that gender,age, family history were independently predisposing factors for COPD.Quality of life was better in non-COPD subjects than in those with COPD,with statistically significant difference.Conclusions Prevalence of COPD was relatively higher in people of rural Tianjin,with gender,age,family history and outdoor air pollution as main risk factors.

ObjectiveTo evaluate the neurobiological characteristics of human histio-amniotic mesenchymal (hAMCs) and effect of hAMCs transplantation into the brain to treat Parkinson's disease(PD) modle mice.MethodsThe expressions of mesenchymal stem cells, neural stem cells, dopaminergic neurons and markers related to neurogenesis such as Vimentin, STRO-1, nestin, CD133, β-tubulin, TH, DAT, Ngn2 and mash-1 in hAMCs were evaluated through immunocytochemical stain; and the mRNA transcriptions of neural stem cell markers, Vimentin and nestin in hAMCs were detected by RT-PCR. The PD model was induced by MPTP(i.p.) in C57BL/6 mice transplanted with hAMCs into the right striatum. The therapeutical effect of hAMCs on PD mice was evaluated by spontaneous movement, rotating bar test and the immunohistochemistry of anti-human chondrosome and TH antibodies in striatum.ResultshAMCs induced by nerve cells culture medium, expressed mesenchymal stem cells, neural stem cells, dopaminergic neurons and other specific markers related to neurogenesis mentioned above. The frequency of spontaneous movement in PD mice was significantly increased(P<0-05), and the time of rotating bar was obviously prolonged(P<0-05) after transplantation with hAMCs.ConclusionhAMCs possess the characteristics of nerve cells after cultured in vitro and can significantly recover the damage of motor function induced by MPTP after transplantation into striatum in PD model mice.

Stroke research and rehabilitation have traditionally focused on the physical and functional impact of a stroke. Less attention has been given to the psychosocial factors associated with this chronic condition. By the few studies that have specifically focused on psychosocial factors in the context of stroke, poststroke depression is demonstrated to significantly influence stroke outcomes. Associations of stroke with psychological symptoms other than depression have rarely been evaluated. This study was aimed to investigate the changes of psychological stress, social support and medication adherence in patients with ischemic stroke in the mainland of China. In this study, 90 patients with hemiplegia one year after first-ever middle cerebral artery infarction (stroke group) in the Zhongnan Hospital of Wuhan University from June 2008 to June 2011 were recruited for interview. Ninety age- and sex-matched normal volunteers (control group) were also examined at the same period. The psychological distress was assessed by the Symptom Checklist 90 (SCL-90), the social support by the Social Support Rating Scale (SSRS), and medication adherence by Morisky's self-reported inventory, respectively. Group differences were analyzed using unpaired-t test and chi-squared test. The results showed that total mean scores of the SCL-90 in the stroke group were higher than those in the control group (P<0.01). Except two dimensions, paranoid ideation and psychoticism, mean scores of the rest dimensions (including somatization, obsession-compulsion, interpersonal sensitivity, depression, anxiety, hostility, and phobic anxiety) of SCL-90 were significantly higher in the stroke group than those in the control group (P<0.05, or P<0.01). The objective support, subjective support, support availability and total social support scores in the stroke group were significantly higher than those in the control group (P<0.05, or P<0.01). Those in the "SCL-90 total scores >150 group" were significantly higher than in the "SCL-90 total scores <100 group" and the "SCL-90 total scores between 100 to 150 group" (P<0.05, or P<0.01). Those in the "SCL-90 total scores between 100 to 150 group" were significantly higher than in the "SCL-90 total scores <100 group" (P<0.05). In 90 patients with ischemic stroke, 26 (28.89%) patients obtained high medication adherence, 47 (52.22%) patients medium medication adherence, and 17 (18.89%) patients low medication adherence, respectively. Among these stroke patients, there were 17 (50.00%) patients with high medication adherence in the "SCL-90 total scores >150 group", 28 (75.67%) patients with medium medication adherence in the "SCL-90 total scores between 100 to 150 group", and 12 (61.16%) patients with low medication adherence in the "SCL-90 total scores <100 group", respectively. There was significant difference in the medication adherence rate among the different SCL-90 scores groups in these stroke patients (P<0.05 or P<0.01). It was led to conclude that ischemic stroke patients one year after hemiplegia have psychological distress, low level of social support and poor medication adherence in the mainland of China. Therefore, it is necessary to mobilize the government, medical institutions and various social support groups to offer psychological interventions to relieve the stress of patients with ischemic stroke, and improve their medication adherence.
Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; psychology ; China ; Female ; Humans ; Infarction, Middle Cerebral Artery ; drug therapy ; psychology ; Male ; Medication Adherence ; statistics & numerical data ; Middle Aged ; Social Support ; Stress, Psychological ; etiology ; psychology ; Surveys and Questionnaires

OBJECTIVETo identify the side effect of all-trans retinoic acid (ATRA), and improve early therapeutic response in patients with acute promyelocytic leukemia (APL).

METHODThe first case of Sweet's syndrome (SS) developed in a APL patient treated with ATRA was reported in mainland of China, and reviewed correlative literature.

RESULTSOnly 14 cases of SS associated with ATRA therapy in APL have been reported in the literature, including the present case. The median age was 49.5 years (9 -84) and 10 were women and 4 men. Of them, SS was restricted to the skin in 10 case, the other 4 muscle, fascia, kidney, and lung were involved. SS appeared after a median of 18 days of ATRA therapy (6 - 34 days). The median WBC count was 7.05 (0.80 - 23.00) x 10(9)/L. Four patients continued with the ATRA therapy without interruption, 13 patients treated with steroids and 12 responded. One patient improved without any treatment. Two cases of SS developed retinoic acid syndromes after ATRA therapy.

CONCLUSIONSweet's syndrome is a rare adverse effect of ATRA, and has similar features with inflammatory or infective dermatosis. The corticosteroids treatment could improve the systemic and cutaneous symptoms. When ATRA therapy was restarted after SS subsided, no recurrence of rashes was observed.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Middle Aged ; Sweet Syndrome ; chemically induced ; Tretinoin ; adverse effects ; therapeutic use

OBJECTIVETo construct human renal cell carcinoma patient-specific full-length antibody library using mammalian cell surface display technique.

METHODSPeripheral blood mononuclear cells (PBMC) were isolated from patients with renal cell carcinoma. The repertoires of kappa light chain (LCkappa) and heavy chain variable region (VH) of antibody were amplified by RT-PCR. The LCkappa and VH libraries were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO10 to construct the renal cell carcinoma patient-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length human antibodies expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of renal cell carcinoma-specific antibody kappa light chain (LCkappa) and heavy chain (IgG1) were constructed. The expression of the full-length human antibodies on the surface of 293T cell was confirmed by flow cytometry. The libraries showed an expressible combinatory diversity of 7.5x10(10).

CONCLUSIONThe expressible antibody library provides a useful platform for screening of renal cell carcinoma-specific antibodies.

Amino Acid Sequence ; Antibodies, Neoplasm ; immunology ; Antibody Specificity ; Carcinoma, Renal Cell ; immunology ; Humans ; Kidney Neoplasms ; immunology ; Molecular Sequence Data ; Peptide Library

OBJECTIVETo investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs.

METHODS① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit.

RESULTS①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002].

CONCLUSION①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.

Bone Marrow Cells ; immunology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokines ; immunology ; Humans ; Immune Tolerance ; Interferon-gamma ; pharmacology ; Mesenchymal Stromal Cells ; immunology ; T-Lymphocytes ; cytology

OBJECTIVETo evaluate the therapeutic effect of QuDu ZengNing Capsule on AIDS.

METHODSQuDu ZengNing Capsule is a capsule containing extract from 4 Chinese medicinal herbs. Totally 1,000 AIDS patients were treated, among them 60 patients were clinically observed weekly. Blood routine tests, liver, heart and kidney function, X-ray, CD4, CD8 cells were examined before and after treatment at 1, 3, 6 month. The patients were treated with 4 capsules t.i.d for 6 months.

RESULTSThe symptoms were improved in most of the patients, the CD4 cells increased from 115.0 to 295.2/ul and the viral load (RNA copies/ml) in most patients reduced markedly or maintained at the same level.

CONCLUSIONThese data indicated that QuDu ZengNing Capsule was effective for treatment of AIDS patients.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; CD4 Lymphocyte Count ; Capsules ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; HIV-1 ; Humans ; Male ; Middle Aged ; Phytotherapy ; Viral Load

OBJECTIVEThis study was aimed to observe the efficacy of autologous stem cell transplantation (ASCT) for adult patients with acute lymphoblastic leukemia (ALL), and investigate related prognostic factors.

METHODSA total of 86 adult ALL patients underwent ASCT in Institute of Hematology and Blood Disease Hospital from November 2001 to January 2012 were followed up. Clinical characteristics and outcomes of all patients were retrospectively analyzed. Survival and univariate prognosis were analyzed by the Kaplan-Meier method and multivariate analysis by COX regression model.

RESULTSOutcomes were assessed in 81 cases, including 47 standard-risk and 34 high-risk patients. 1-, 3-, 5-, and 10-year leukemia-free survival (LFS) of standard-risk patients were (82.3±5.7)%, (76.9±6.5)%, (74.1±6.8)%, (67.4±8.9)% respectively,and relapse rates (RR) were as of (13.6±5.2)%, (21.6±6.4)%, (24.5±6.8)%, (31.3±9.0)% respectively. 1-, 3-, 5-, and 10-year LFS of high-risk patients were (55.8±8.9)%, (39.8±9.3)%, (39.8±9.3)%, (39.8±9.3)% respectively, and relapse rates (RR) were (38.8±9.2)%, (56.4±10.0)%, (56.4±10.0)%, (56.4±10.0)% respectively. T-ALL, white blood cell count(WBC) more than 30×109/L when first visited, increased LDH, positive fusion gene of TCR and bone marrow transplantation were the adverse prognostic factors. Multivariate analysis showed bone marrow transplantation was an independent adverse prognostic factor.

CONCLUSIONASCT was a choice for adult ALL patients when suitable donors were unavailable.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Prognosis ; Retrospective Studies ; Risk Factors ; Transplantation, Autologous ; Young Adult