Clinical and laboratory characteristics of 11 cases of chronic B lymphocyte proliferation disease without lymphoid proliferation / 肿瘤研究与临床

Objective To explore the clinical and laboratory characteristics of chronic B lymphocyte proliferation disease (B-CLPD) without typical lymphoid proliferation. Methods The clinical records of patients with B-CLPD only characterized by pancytopenia form January 2007 to March 2016 in hematology department of Xinjiang Uygur Autonomous Region People ' s Hospital were collected, and the cell morphology, bone marrow pathology, cytogenetics and molecular characteristics were retrospectively analyzed. Results The median age of 11 patients was 68 years old. The lymphocyte ratio of peripheral blood smears in all patients increased in different level (0.36-0.68), but absolute lymphocyte count was normal or decreased (0.59×109-1.99×109). Lymphocyte-like plasma cell or small numbers of plasma cell can be seen in the bone marrow smears of 4 cases and lymphocytes with irregular burr-like protrusions were observed in 2 cases while there were no characteristic morphological changes in remained 5 cases. Immunophenotypical analysis showed that all patients expressed CD19, CD20, CD22, SmIg, not expressed CD5, CD10 which with scores of 0-2 according to chronic lymphocytic leukemia (CLL) points system;CD103, CD11C, CD25 and FMC7 were highly expressed in 2 cases while there were no characteristic expression in remaining cases. There were no abnormal karyotypes observed from the conventional cytogenetic and fluorescence in situ hybridization (FISH) analysis (both of IgH/CCND1, bcl-2/IgH were negative) in all patients. 8 patients were found IgH gene rearrangement, MYD88L256P and BRAF V600E was positive in 5 cases and 2 cases respectively. 5 cases were diagnosed as Waldenstrom macroglobulinemia, 3 cases were B-CLPD, 2 cases were hairy cell leukemia, 1 case was nodal marginal zone B-cell lymphoma after comprehensive analysis of their clinical and laboratory data. Conclusion Even if there are no increased peripheral blood lymphocytes in pancytopenia patients, it is necessary to perform bone marrow smears, immunophenotyping, IgH gene rearrangement, cytogenetics and other molecular laboratory tests to exclude B-CLPD, and reduce misdiagnosis.

Preparation and clinical application of a novel monoclonal antibody against Mycobacterium tuberculosis culture filtrate protein 10 / 中华临床感染病杂志

Objective To prepare a novel monoclonal antibody (mAb) that specifically against Mycobacterium tuberculosis culture filtrate protein 10 (CFP-10).Methods The BALB/c mice were immunized by a peptide with 14 amino acids (aa residues 53 to 66) of CFP-10,and then the splenocytes of mice were fused with myeloma cell line SP2/0.The resultant fused cells were subjected to screening culture,enzyme linked immunosorbent assay (ELISA) assay and subcloning by limited dilution to establish hybridoma cell lines of stable secreting anti-the peptide of CFP-10 antibody.The antibody was purified,and its isotypes were analyzed.Then,the antibody was further evaluated by Western blotting,immunoprecipitation and ELISA in 38 culture supernatant samples of Mycobacterium tuberculosis,20 culture supernatant samples of non-Mycobacterium tuberculosis,32 samples of tuberculous pleural effusion,24 samples of non-tuberculous pleural effusion,and 20 serum samples from healthy controls.Results The isotype of the mAb against the specific peptide of CFP-10 was an IgG1 with κ chain,and it was applicable for Western blotting and immunoprecipitation analysis.ELISA quantitative test showed that the sensitivity and specificity for diagnosis of Mycobacterium tuberculosis were 78.6％ (55/70) and 92.2％ (59/64),respectively.Conclusion The mAb generated against the specific peptide of CFP-10 is high in sensitivity and specificity,and it might be used in the early diagnosis of tuberculosis.

The trend of development of clinical immunology in laboratory medicine / 中华检验医学杂志

With the rapid development of the individual medical care in the 21st century,based on the quick advancement of the preclinical immunology as well as the relevant bioscience technology,the application of clinical immunological diagnostic inspection has been involved in multiple diseases,including the investigation of the pathology,diagnosis,selection of the treatment regime and the evaluation of the therapy efficacy.Along with the further research,novel means of the treatment and the inspection technology have been to be utilized or explored.The focus should be concentrated on the utilization of the new technologies,new ways of the clinical immunological inspection,improving the fabric of the knowledge and the application ability of the professionals in this major,in order to provide effective service to the clinical medical care and the health of the patients.

Prevalence of allergen sensitization, most important allergens and factors associated with atopy in children / Prevalencia de sensibilizacao a alergenos, alergenos mais relevantes e fatores associados com a atopia em criancas

CONTEXT AND OBJECTIVE: Knowledge of the profile of allergen sensitization among children is important for planning preventive measures. The objective of this study was to assess the prevalence and profile of sensitization to inhaled allergens and food among children and adolescents in an outpatient population in the city of Palmas. DESIGN AND SETTING: Cross-sectional study at outpatient clinics in Palmas, Tocantins, Brazil. METHODS: Ninety-four patients aged 1-15 years who were attending two pediatric outpatient clinics were selected between September and November 2008. All of the subjects underwent clinical interviews and skin prick tests. RESULTS: A positive skin prick test was observed in 76.6% of the participants (72.3% for inhalants and 28.9% for food allergens). The most frequent allergens were Dermatophagoides pteronyssinus (34%), cat epithelium (28.7%), dog epithelium (21.3%), Dermatophagoides farinae (19.1%), Blomia tropicalis (18.1%), cow's milk (9.6%) and grasses (9.6%). A positive skin prick test correlated with a history of atopic disease (odds ratio, OR = 5.833; P = 0.002), a family history of atopic disease (OR = 8.400; P < 0.001), maternal asthma (OR = 8.077; P = 0.048), pet exposure (OR = 3.600; P = 0.012) and cesarean delivery (OR = 3.367; P = 0.019). CONCLUSION: Dermatophagoides pteronyssinus was the most frequent aeroallergen and cow’s milk was the most prevalent food allergen. There was a positive correlation between a positive skin prick test and several factors, such as a family history of atopic disease, maternal asthma, pet exposure and cesarean delivery. .

CONTEXTO E OBJETIVO: O conhecimento sobre o perfil da sensibilização a alérgenos em crianças é importante para o planejamento de medidas preventivas. O objetivo deste estudo foi avaliar a prevalência e perfil de sensibilização a alérgenos inalados e alimentares em crianças e adolescentes em uma população ambulatorial na cidade de Palmas. TIPO DE ESTUDO E LOCAL: Estudo transversal em unidades ambulatoriais em Palmas, Tocantins, Brasil. MÉTODOS: Foram selecionados 94 pacientes com idades entre 1 a 15 anos em 2 ambulatórios de pediatria entre setembro e novembro de 2008. Todos os sujeitos foram submetidos a entrevistas clínicas e testes cutâneos de puntura. RESULTADOS: Um teste cutâneo de puntura positivo foi observado em 76,6% dos participantes (72,3% para inalantes, 28,9% para alérgenos alimentares). Os alérgenos mais frequentes foram Dermatophagoides pteronyssinus (34%), epitélio de gato (28,7%), epitélio de cão (21,3%), Dermatophagoides farinae (19,1%), Blomia tropicalis (18,1%), leite de vaca (9,6%) e gramíneas (9,6%). Um teste cutâneo de puntura positivo foi relacionado à história de doença atópica (razão de chances RC = 5,833, P = 0,002), história familiar de atopia (RC = 8,400, P < 0,001), asma materna (RC = 8,077, P = 0,048), exposição a animal de estimação (RC = 3,600, P = 0,012) e parto cesáreo (RC = 3,367, P = 0,019). CONCLUSÃO: Dermatophagoides pteronyssinus foi o aeroalérgeno mais prevalente e, dentre alérgenos alimentares, o leite de vaca. Houve correlação positiva entre o teste cutâneo e alguns fatores, como história familiar de atopia, asma materna, exposição a animais domésticos e parto cesáreo. .

A study on the immunologic techniques experiment teaching of higher vocational education / 中华医学教育探索杂志

The student majoring in Medical Biological Technique will be mainly engaged in the practical work of biological technique industry after graduation in the future.In order to bind the theories on the practical biological techniques in designs of contents and roundly improve students' practical ability in classes as well as enriching the communication among students,our college offers an immunologic techniques experiment classes with 36 hours per semester,which has also undergone a reasonable project teaching innovation. This proved to result in a satisfactory outcome in improving the students' practical a

Identification and characterization of single chain antibody of cancer stem cell surface marker CD133 scFv CD133 / 肿瘤研究与临床

Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.

Advance of detection of colorectal cancer micrometastasis / 肿瘤研究与临床

With the changes of lifestyle, the incidence of colorectal cancer is increasing year by year.Though the surgical techniques and adjuvant therapy means are continuously improving, the overall prognosis of patients is still poor. Micrometastasis of colorectal cancer is the main reason leading to recurrence and metastasis. Timely detection of micrometastasis is meaningful to accurately determine the condition, formulate reasonable treatment and improve survival time. With the development of medical detection and sophisticated means, the micrometastasis of coloroctal cancer detection has become the research hot spots. In this paper, the current status of micrometastasis of colorectal cancer detection methods, detection means and choice of marker are introduced.

The preparation of anti-hnRNP A2/B1 polyclonal antibody and its application in non-small cell lung cancer / 肿瘤

Objective: To prepare anti-hnRNP A2/B1 polyclonal antibody and determine its clinical significance in diagnosis and prediction of non-small cell lung cancer and analyze the relationship between the expression of hnRNP A2/B1 and pathological features. Methods: HnRNP A2/B1 expression was induced by IPTG. The rabbit-anti-human hnRNP A2/B1 polyclonal antibodies were prepared and purified. The specimens were isolated from 45 cases of non small cell lung cancer tissues and paracancerous tissues and 16 cases of lung inflammatory pseudotumor tissues and embedded in paraffin. hnRNP A2/B1 expression was detected by immunohistochemical method using rabbit anti-human hnRNP A2/B1 polyclonal antibodies. The results were compared with imported anti-hnRNPA2/B1 monoclonal antibody to observe the clinical value of self-made polyclonal antibodies. Results: The rabbit anti-human hnRNPA2/B1 polyclonal antibody is successfully generated. The positive rate of hnRNP A2/B1 expression detected by anti-hnRNP A2/B1 polyclonal antibody was significantly higher in lung cancer tissues than that in paracancerous tissues (62.2% vs 40.0%, P = 0.035) and lung inflammatory pseudotumor tissues(62.2% vs 31.3%, P = 0.033). The overexpression of hnRNP A2/B1 was positively related with age and smoking and negatively related with differentiation degree. But it was not related to tumor histological classification, gender, lymph node metastasis, TNM stage, and distant metastasis. (4) The difference in survival time was significant between the two groups. hnRNP A2/B1-positive patients had shorter survival time than hnRNP A2/ B1-negative patients (P = 0.048). Conclusions: The rabbit anti-human hnRNP A2/B1 polyclonal antibody is successfully generated. Overexpression of hnRNP A2/B1 in lung cancer tissues has clinical values to a certain degree for diagnosis and prediction of the prognosis of lung cancer.

Expression and its role of aquaporin in the middle ear of the guinea pigs / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the expression pattern of AQP1,AQP4,AQP5 in the normal middle ear in guinea pigs and its roles in the water homeostasis in the middle ear cavity.METHODS Reverse transcription polymerase chain reaction(RT- PCR),Western blotting and immunohistochemical staining were used to detect AQP1,AQP4,AQP5 in mucosa of the bullae of normal guinea pigs.RESULTS Reverse transcription-polymerase chain reaction and immunoblot analyses revealed that mRNAs encoding AQP1,AQP4,AQP5 were expressed in middle ear membrane of the guinea pigs.AQP1,AQP4 was also detected as 28-kDa,33-kDa proteins in middle ear cavity of guinea pig.Immunohistochemical staining showed that only AQP1 located at capillary endothelial cells and fibroblasts of the submucous layer,as well as at flat and cubical epithelial cells.CONCLUSION The results suggest that AQP1,AQP4,AQP5 can express in the normal middle ear cavity of guinea pigs and may play a vital role in the water transmission in the middle ear.The air-filled middle ear cavity may result from the working of AQP1,AQP4,AQP5.

Improved immunodetection of human papillomavirus E7

Human papillomavirus E7 (HPV E7) is a viral oncoprotein that plays an important role in cervical carcinogenesis through binding with retinoblastoma protein (Rb). Inactivation of Rb by E7 is necessary but not sufficient for cellular transformation, suggesting other protein-protein interactions are required for E7-mediated cellular transformation aside from the interaction with Rb. However, studies on the oncogenic function of HPV E7 have been limited by its poor immunoreactivity. In this report, we show that the fixation of purified recombinant HPV E7 on blotted nitrocellulose membrane with glutaldehyde markedly enhanced the immunoreactivity of HPV E7 protein. Using HeLa and Caski cell line which are infected with HPV 18 and HPV 16, respectively, we demonstrated that native HPV E7 proteins also could be detected by this method. These results therefore can provide the experimental conditions for detection of HPV E7 proteins with greater sensitivity and may help to analyze E7 functions.

Procedimientos para la detección e identificación de anticuerpos eritrocitarios / Procedures for the detection and identification of red cell antibodies

En Inmunohematología se ha desarrollado una amplia gama de procederes de detección e identificación de anticuerpos eritrocitarios in vitro, por lo cual se realiza una revisión de técnicas y métodos empleados con este objetivo, como son el método que utiliza eritrocitos pretratados con enzimas proteolíticas y las técnicas de Polibreno, que utiliza solución de baja fuerza iónica (LISS), la de antiglobulina indirecta, la de aglutinación en gel, la inhibición de la aglutinación, la hemólisis y la adherencia de eritrocitos en fase sólida. Se abordan los problemas que afectan a la reacción de aglutinación entre el antígeno y el anticuerpo; para una mejor comprensión la reacción de aglutinación se subdivide en su primera y segunda etapa. En la primera etapa los factores que se analizan son concentración de antígeno y anticuerpo, pH, temperatura, fuerza iónica y tiempo de incubación; en la segunda etapa la característica del anticuerpo, localización y número de sitios antigénicos, fuerzas que mantienen la distancia entre los eritrocitos, uso de la albúmina bovina, uso de enzimas, efecto de dosis y efecto de moléculas con carga positiva.

Awide range of procedures for the detection and identification of red cell antibodies in vitro has been developed in Immunohematology. Therefore, it is made a review of the techniques and methods used with this aim, such as the method using erythrocytes pretreated with proteolytic enzimes and the techniques of Polibreno that utilize low ionic force solution (LIFS), the indirect antiglobulin test, the gel agglutination test, the agglutination inhibition test, hemolysis and the solid phase erythrocyte adherence test. The problems affecting the agglutination reaction between the antigen and the antibody are also dealt with. The agglutination reaction is subdivided into its first and second phase for a better understanding. In the first phase, the antigen and antibody concentration, pH, temperature, ionic force and incubation time are analyzed. The characteristic of the antibody, localization and number of antigenic sites, forces maintaining the distance among the erythrocytes, use of bovine albumin, use of enzimes, dose effect and effect of molecules with positive charge are studied in the second phase.

Tumor de células granulares de la región anal: Presentación de 1 caso / Tumor of granular cells of the anal region. Presentation of 1 case

Se informa un caso de tumor de células granulares (mioblastoma) de la región anal, en una mujer de 54 años de edad que acudió al Hospital Clinicoquirúrgico Docente "Joaquín Albarrán" por "quiste palpable en las márgenes del ano. El tratamiento que se realizó fue la resección local del tumor. Se analizaron los patrones histopatológicos e inmunohistoquímicos, los cuales se correspondieron con los reportados en la literatura médica hasta el presente(AU)

The authors present a case of granular cell tumor (myoblastoma) of the anal region in a woman aged 54, who was attended at "Joaquin Albarrán" Clinical Surgical Hospital due to a palpable "cyst" in the anal edges. The treatment applied consisted in local resection of the tumor. Histopathologic and immunohistochemical patterns were analyzed and they corresponded to those reported in medical literature(AU)

Study on the role of interleukins in children with asthma / 中华检验医学杂志

Objective To study the role of IL 12 and IL 13 mRNA in children with asthma. Methods Use of semi quantitative RT PCR, IL 12 and IL 13 mRNA in peripheral blood mononuclear cell (PBMC), as well as total IgE in serum from children with asthma, which is in the period of acute phase, were detected. Results Compared with control group, The expression level of IL 12 mRNA were decreased and that of IL 13 mRNA were increased in asthmatic children; The sicker the patient was, the lower expression of IL 12 mRNA, the higher expression of IL 13 mRNA; No matter how the IgE level was, there was significantly different between the expression of IL 12 and IL 13 mRNA. Conclusion IL 12 and IL 13 may be one of the factors causing bronchial chronic inflammation.