Research progresses of smart responsive 19F magnetic resonance nanomolecular imaging probes in diagnosis of tumors / 中国医学影像技术

Using smart responsive nanomolecular probes can realize tumor targeted imaging with rapid response, sensitive diagnosis and precise treatment under the guidance of imaging on detection of probe distributions and localization of tumors. MRI, especially 19F-MRI, has high sensitivity and low background noise. In recent years, smart responsive 19F-MR nanomolecular imaging probes had been applied for detecting changes of pH, enzyme activity, reducing microenvironment, hypoxia, light and thermo-responsive aspects. The research progresses of smart responsive 19F magnetic resonance nanomolecular imaging probes in diagnosis of tumors were reviewed in this article.

Research advances of molecular probes for early diagnosis of liver fibrosis / 中国医学影像技术

Early liver fibrosis is reversible and curable. Therefore, early diagnosis and staging of liver fibrosis are of great clinical significance. Targeting at the abnormal expression molecules in development of liver fibrosis, molecular probes are constructed as precise and non-invasive methods to integrate the information of human physiological and pathological metabolism, which is beneficial to early and specific diagnosis of fibrosis. The advances in molecular probes for early diagnosis and staging of liver fibrosis were reviewed in this article,

Use and evaluation of a line probe assay in patients with tuberculosis in Peru: 2011 and 2013 / Utilización y evaluación de un ensayo con sondas en línea en los pacientes con diagnóstico de tuberculosis en el Perú del 2011 al 2013

ABSTRACT Objective To determine the use and performance of a line probe assay (LPA) compared with conventional culture and drug sensitivity testing (CDST) in patients registered with tuberculosis (TB) under routine program conditions in Peru in 2011–2013. Methods This was a descriptive, operational research, cross-sectional study of sputum specimens from patients with smear-positive pulmonary TB and mycobacterial cultures from patients with smear-negative or positive TB. Drug resistance to rifampicin and/or isoniazid detected by LPA was compared to CDST. Sensitivity, specificity, and predictive values were calculated and reliability for detecting drug resistance was assessed through kappa coefficient, with values 0.61–0.80 showing substantial correlation, and 0.81 or above showing almost-perfect correlation. Results In 2011–2013, there were 16 169 LPA tests performed, with the proportion of TB patients receiving the test increasing from 3.2% to 30.2%. In all, 2 905 LPA test results were compared to CDST. For LPA in sputum specimens, sensitivity for rifampicin was 92%; isoniazid, 94%; and MDR-TB, 88%; while specificity for rifampicin was 92%; isoniazid, 92%; and MDR-TB, 95%. For LPA in mycobacterial cultures, sensitivity for rifampicin was 95%; isoniazid, 96%; and MDR-TB, 90%; while specificity for rifampicin was 85%; isoniazid, 91%; and MDR-TB, 94%. Kappa coefficients were at 0.81 or above for all comparisons of LPA with CDST using sputum specimens and cultures, except for isoniazid in cultures, which was at 0.79. Conclusions This study suggests that LPA is a reliable and rapid screening test for drug-resistant TB and should be considered suitable for routine use and scale up in Peru.

RESUMEN Objetivo Definir la utilización de un ensayo con sondas en línea y evaluar su desempeño, en comparación con el método convencional de cultivo y antibiograma, en los pacientes registrados con tuberculosis en condiciones programáticas en el Perú del 2011 al 2013. Métodos Investigación operativa descriptiva con un estudio transversal de las muestras de esputo de los pacientes con diagnóstico de tuberculosis pulmonar y baciloscopia positiva y de los cultivos de micobacterias de los pacientes con tuberculosis y baciloscopia positiva o negativa. La farmacorresistencia a la rifampicina, la isoniacida o a ambas, detectada mediante el ensayo con sondas en línea, se comparó con los resultados obtenidos por el método de cultivo y antibiograma. Se calculó la sensibilidad, la especificidad y los valores predictivos del ensayo con sondas en línea y se evaluó su fiabilidad en la detección de la farmacorresistencia mediante el coeficiente k, cuyos valores de 0,61 a 0,80 correspondían a una fuerte correlación y los valores de 0,81 o superiores reflejaban una correlación casi perfecta. Resultados Del 2011 al 2013 se practicaron 16 169 ensayos con sondas en línea, y la proporción de pacientes con diagnóstico de tuberculosis en quienes se practicaba aumentó de 3,2% a 30,2%. En total, se compararon 2 905 resultados del ensayo molecular con el método convencional. En las muestras de esputo, el ensayo molecular ofreció una sensibilidad de 92% para la resistencia a la rifampicina, 94% a la isoniacida y 88% para la tuberculosis multirresistente; su especificidad fue 92% con respecto a la rifampicina, 92% a la isoniacida y 95% a la tuberculosis multirresistente. En los cultivos de micobacterias, el ensayo con sondas en línea mostró una sensibilidad de 95% para la resistencia a la rifampicina, 96% a la isoniacida y 90% para la tuberculosis multirresistente; la especificidad fue 85% para la rifampicina, 91% para la isoniacida y 94% para la tuberculosis multirresistente. El coeficiente k fue 0,81 o superior en todas las comparaciones del ensayo molecular con el método tradicional cuando se usaron muestras de esputo y cultivo, excepto con la isoniacida en cultivo, cuyo coeficiente fue 0,79. Conclusiones Los resultados del presente estudio indican que el ensayo con sondas en línea constituye una prueba de detección fiable y rápida para la tuberculosis multirresistente, y se debe considerar apropiada su utilización en la práctica de rutina y la ampliación de su empleo en el Perú.

Use of amplified Mycobacterium tuberculosis direct test in respiratory samples from HIV-infected patients in Brazil / Utilização do amplified Mycobacterium tuberculosis direct test em amostras respiratórias de pacientes HIV positivos no Brasil

OBJECTIVE: To compare the accuracy of the amplified Mycobacterium tuberculosis direct (AMTD) test with reference methods for the laboratory diagnosis of tuberculosis in HIV-infected patients. METHODS: This was a study of diagnostic accuracy comparing AMTD test results with those obtained by culture on Löwenstein-Jensen (LJ) medium and by the BACTEC Mycobacteria Growth Indicator Tube 960 (BACTEC MGIT 960) system in respiratory samples analyzed at the Bioassay and Bacteriology Laboratory of the Oswaldo Cruz Foundation Evandro Chagas Clinical Research Institute in the city of Rio de Janeiro, Brazil. RESULTS: We analyzed respiratory samples collected from 118 patients, of whom 88 (74.4%) were male. The mean age was 36.6 ± 10.6 years. Using the AMTD test, the BACTEC MGIT 960 system, and LJ culture, we identified M. tuberculosis complex in 31.0%, 29.7%, and 27.1% of the samples, respectively. In comparison with LJ culture, the AMTD test had a sensitivity, specificity, positive predictive value, and negative predictive value of 87.5%, 89.4%, 75.7%, and 95.0%, respectively, for LJ culture, whereas, in comparison with the BACTEC MGIT 960 system, it showed values of 88.6%, 92.4%, 83.8%, and 94.8%, respectively. CONCLUSIONS: The AMTD test showed good sensitivity and specificity in the population studied, enabling the laboratory detection of M. tuberculosis complex in paucibacillary respiratory specimens. .

OBJETIVO: Comparar a acurácia do teste amplified Mycobacterium tuberculosis direct (AMTD) com métodos de referência para o diagnóstico laboratorial de tuberculose em pacientes HIV positivos. MÉTODOS: Estudo de acurácia diagnóstica comparando os resultados do teste AMTD com os de cultura em Löwenstein-Jensen (LJ) e de BACTEC Mycobacteria Growth Indicator Tube 960 (sistema BACTEC MGIT 960) em amostras respiratórias analisadas no Laboratório de Bacteriologia e Bioensaios do Instituto de Pesquisa Clínica Evandro Chagas da Fundação Oswaldo Cruz, no Rio de Janeiro (RJ). RESULTADOS: Foram analisadas amostras respiratórias de 118 pacientes, dos quais 88 (74,4%) eram do sexo masculino. A média de idade foi de 36,6 ± 10,6 anos. O complexo M. tuberculosis foi identificado em 31,0%, 29,7% e 27,1% das amostras através do teste AMTD, sistema BACTEC MGIT 960 e LJ, respectivamente. Na comparação com a cultura em LJ, o teste AMTD apresentou sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 87,5%, 89,4%, 75,7% e 95,0%, respectivamente, enquanto na comparação com o sistema BACTEC MGIT 960, os valores foram de 88,6%, 92,4%, 83,8% e 94,8%, respectivamente. CONCLUSÕES: O teste AMTD mostrou boa sensibilidade e especificidade na população estudada, possibilitando a detecção laboratorial do complexo M. tuberculosis em espécimes respiratórios paucibacilares. .

Advances in follicle stimulating hormone receptor and its targeted drugs / 肿瘤

Follicle stimulating hormone receptor (FSHR) belongs to the G protein-coupled receptor family and is only low-expressed in reproductive organs of mammals. Recent studies have shown that FSHR is overexpressed in vascular endothelium of multiple types of human tumors including prostate cancer, breast cancer, liver cancer and kidney cancer, etc, and it plays an important role in the progression of cancer. Thus, FSHR is a potential target for diagnosis and treatment of tumors. Development of drugs targeting FSHR is beneficial to diagnosis, targeted therapy, selection of individualized drug therapy and prognosis of tumors. This paper summarizes the advances in FSHR and its specific ligands.

Molecular inversion probe-based single base mutations of hepatitis B virus resistance gene analysis / 中华检验医学杂志

Objective To establish a molecular inversion probe ( MIP) method for detection of single base drug-resistance mutation in Hepatitis B virus ( HBV) gene.Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy.Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations , the detection sensitivity of MIP was determined as 1 nmol/L.The results of MIP were consistent with that of sequencing method in detection of the clinical samples.Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.

Multidrug-Resistant Tuberculosis Presenting as Miliary Tuberculosis without Immune Suppression: A Case Diagnosed Rapidly with the Genotypic Line Probe Assay Method / 결핵및호흡기질환

Miliary tuberculosis (TB) is a rare extrapulmonary form of TB, and there have been only two reports of miliary TB associated with infection with multidrug-resistant (MDR)-TB pathogen in an immunocompetent host. A 32-year-old woman was referred to our hospital because of abnormal findings on chest X-ray. The patient was diagnosed with MDR-TB by a line probe assay and was administered proper antituberculous drugs. After eight weeks, a solid-media drug sensitivity test revealed that the pathogen was resistant to ethambutol and streptomycin in addition to isoniazid and rifampicin. The patient was then treated with effective antituberculous drugs without delay after diagnosis of MDR-TB. To the best of our knowledge, this is the first case of miliary TB caused by MDR-TB pathogen in Korea.

Construction of transcriptome profiling of cancerous parenchyma cells using laser capture microdissection and Gene-Chip technology / 肿瘤

Objective: To construct transcriptome profiling of parenchyma cell using laser capture microdissection (LCM) combined with Gene-Chip technology. Methods: We obtained cancerous parenchyma cells from frozen sections of tumor tissues by using LCM. RNA was extracted from the cells. aRNA was obtained after linear amplification, and then converted to cRNA probe, finally hybridized with Affymetrix HG-U133. Plus 2.0 Gene-Chip for construction of full genome transcriptome profiling. Results: About 3 mm2 cancerous colon cells were obtained from each of 5 colon cancer specimens, respectively. The quantity of total RNA extracted from cancerous colon cells ranged from 118.4 to 300.3 ng. We obtained 7 υg of aRNA after linear amplification from 100 ng mixed RNA. The quality control was guided by the operating instruction standard of Gene-Chip. Totally 18 205 transcripts were presented representing 15 276 genes. Conclusion: The combination of laser capture microdissection and Gene-Chip technology could be used for construction of transcriptome profiling of purified cancerous parenchyma cells.

Detection of the immunologic rejection after xeno-islet transplantation:a study by MR imaging enhanced with superparamagnetic iron oxide marking CD4+T cell antibody / 中华放射学杂志

Objective To evaluate the feasibility of the diagnosis of the early immunologic rejection after xeno-islet transplantation by MR imaging enhanced with superparamagnetic iron oxide(SPIO)marking CD4+T cell antibody.Methods Two thousand neonatal porcine islets(NPI)were transplanted under the left renal capsule of BALB/C nude mice.When the grafts could be observed bv MRI.107 human PBMC was intraperitoneal injected to nude mouse models to reconstitute the human immunologic system,20 mice were reconstituted.Before and 3,7,14 days after reconstitution of human immunologic system on BALB/C nude mice,MRI imaging Was performed half an hour after intravenous injection of nano-immunomagnetic beads via vena caudalis to observe the grafts'MRI signal.BALB/C nude mice were sacrificed after MRI scanning immediately,the histopathologic examination was assessed on grafts,the results were compared with MRI results.And calculate the sensitivity,specificity,Youden index number and coincidence of the MRI for immunologic rejection.Results Grafts can be observed bv MRI 3 weeks after islet cell transplantation (before immunologic rejection modeling),there is no abnormal MRI signal detected in nude mice'graft region after mierobeads injected.Seven days after building of immunologic rejection model,MRI hypo-signal in graft site is shown in the T2 WI sequence after nano-bioprober injected.Histopathologic assessments were employed on grafts in nude mice immediately(HE and immunohistochemistry staining),the results shown that there are a lot of T lymphocyts infiltrated in graft region.implying the occurrence of immunologic rejection.And the sensitivity,specificity,Youden index number and coincidence is:(72.96±0.24)%,100%,0.73±0.24,(88.46±0.13)%respectively.The correct Kappa between the MRI and the imunohistochemistry staining was 0.76.Conclusion The cellular immunological rejection to xeno-islet grarts can be assessed with nano-bioprobe with anti-CD4+ antibody MR imaging,real time,and noninvasively.

Establishment of a rapid quantitative method for Bacillus anthracis / 中华检验医学杂志

Objective To establish a rapid specific quantitative assay for Bacillus anthracis detection. Methods According to the principle of complex probe quantitative assay, the primers and quantitative probes targeted at chromatosome DNA rpoB were designed and applied to detect Bacillus anthracis. The influence factor of quantitative PCR were determined. Results The optimal system of this method was aquired: the length of quenching probe is 15mer,the ratio of fluorescent probe to quenching probe is 1/2 and the concentrtion of Mg 2+ is 3 mmol/L.The sensitivity of this assay for Bacillus anthracis is 10 3 copies. It can distinguish Bacillus anthracis from other closely related Bacillus. Conclusion The method can rapidly quantitatively detect the Bacillus anthracis with high sensitivity and specificity, it can be applied to clinical diagnosis.