RESUMO
Columnaristerol A (1), a rare natural 19-norsterol possessing a 10ß-hydroxy group was isolated from the Formosan octocoral Nephthea columnaris, and its structure was elucidated by spectroscopic methods. Sterol 1 was found to be a cytotoxic agent that exhibited in vitro moderate cytotoxic activity against MOLT-4 and SUP-T1 human leukemia-lymphoma cell lines.
Assuntos
Antozoários/metabolismo , Noresteroides/química , Noresteroides/farmacologia , Esteróis/química , Esteróis/farmacologia , Animais , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Espectroscopia de Prótons por Ressonância Magnética , Relação Estrutura-Atividade , TaiwanRESUMO
BACKGROUND: The shell of Haliotis diversicolor, or shijueming (SJM), is a type of traditional Chinese medicine. The SJM has appeared in historical records as early as the third and fourth centuries. Historical records have revealed that SJM had mainly been used to treat eye diseases. After the Qing Dynasty (1757), records had emerged, detailing the use of SJM for treating skin injuries, particularly for treating poorly managed ulcers or traumatic wounds. Furthermore, in our anti-inflammation-screening system, SJM significantly inhibited the expression of pro-inflammatory proteins. Previous studies have yet to adopt an animal model to verify the phenomenon and described in the historical records regarding the efficacy of SJM in promoting wound healing. Besides, the mechanism of wound healing effect of SJM is also not clear. METHODS: This study applied in vitro and in vivo models, tissue section analysis, and western blotting to evaluate the effect of SJM on wound healing. The RAW 264.7 cells were used in anti-inflammatory activity assay and phagocytic assay. Male Wistar rats were used to evaluate the effect of SJM on burn injury healing. A copper block (2 × 2 cm, 150 g) preheated to 165 °C in a dry bath was used to contact the skin area for 10 s, thus creating a full-thickness burn injury. The results were analyzed by hematoxylin and eosin staining, picrosirius red staining and Western blotting. RESULTS: The results revealed that in the in vitro model, the presence of SJM decreased the inducible nitric oxide synthase (iNOS) expression and enhanced the functions of macrophages. The results of the rat burn injury model revealed that SJM decreased neutrophil infiltration, promoted wound healing, thus increasing the collagen I content and promoting the expression of transforming growth factor-beta 1 (TGF-ß1) protein. We speculate that the effect and mechanism of SJM on promoting wound healing is related to macrophage activation. In the inflammation phase, SJM alleviates inflammation by inhibiting iNOS expression and removing neutrophils through phagocytosis. Furthermore, SJM induces the secretion of TGF-ß1, converting collagen during the tissue remodeling phase. CONCLUSIONS: According to our review of relevant literature, this is the first study that applied an evidence-based method to verify that SJM alleviates inflammation, enhances phagocytosis, and triggers wound healing after burn injury. The study findings reveal that SJM provides a promising therapeutic option for treating burn injury.
Assuntos
Exoesqueleto/química , Queimaduras/tratamento farmacológico , Gastrópodes/química , Cicatrização/efeitos dos fármacos , Animais , Linhagem Celular , Masculino , Medicina Tradicional Chinesa , Óxido Nítrico Sintase Tipo II/metabolismo , Fagocitose/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
BACKGROUND: Syphilis is an infectious disease caused by Treponema pallidum that can invade the central nervous system, causing encephalitis. Few cases of anti-N-methyl-D-aspartate receptor autoimmune encephalitis (AE) secondary to neurosyphilis have been reported. We report a neurosyphilis patient with anti-γ-aminobutyric acid-B receptor (GABABR) AE. CASE SUMMARY: A young man in his 30s who presented with acute epileptic status was admitted to a local hospital. He was diagnosed with neurosyphilis, according to serum and cerebrospinal fluid (CSF) tests for syphilis. After 14 d of antiepileptic treatment and anti-Treponema pallidum therapy with penicillin, epilepsy was controlled but serious cognitive impairment, behavioral, and serious psychiatric symptoms were observed. He was then transferred to our hospital. The Mini-Mental State Examination (MMSE) crude test results showed only 2 points. Cranial magnetic resonance imaging revealed significant cerebral atrophy and multiple fluid-attenuated inversion recovery high signals in the white matter surrounding both lateral ventricles, left amygdala and bilateral thalami. Anti-GABABR antibodies were discovered in CSF (1:3.2) and serum (1:100). The patient was diagnosed with neurosyphilis complicated by anti-GABABR AE, and received methylprednisolone and penicillin. Following treatment, his mental symptoms were alleviated. Cognitive impairment was significantly improved, with a MMSE of 8 points. Serum anti-GABABR antibody titer decreased to 1:32. The patient received methylprednisolone and penicillin after discharge. Three months later, the patient's condition was stable, but the serum anti-GABABR antibody titer was 1:100. CONCLUSION: This patient with neurosyphilis combined with anti-GABABR encephalitis benefited from immunotherapy.
RESUMO
Funiu Mountains are located in a transition region between warm temperate zone and northern subtropical region, where a variety of plant species are distributed with sensitive response to climate change. Their response characteristics to climate change are still unclear. We developed the basal area increment (BAI) index chronologies of Pinus tabuliformis, P. armandii, and P. massoniana in the Funiu Mountains to examine their growth trend and their sensitivity to climatic change. The results showed that the BAI chronologies gave a clue that the three conife-rous species had similar radial growth rate. The large Gleichlufigkeit (GLK) indices among the three BAI chronologies also indicated that the three species had a similar growth trend. Results of correlation analysis showed that the three species also had similar response to climatic change to a certain extent. Radial growth of all the three species was significantly positively correlated with the total monthly precipitation in December of previous year and June of the current year, but negatively correlated with the precipitation in September and the mean monthly temperature in June of the current year. There were some differences in the responses of the three coniferous to climate change. P. massoniana had a significant negative correlation with the mean temperature in March, and a significant positive correlation with the precipitation in March, while P. armandii and P. massoniana were affected negatively by the maximum temperature in August. Results of the moving correlation analysis showed that the three coniferous species had some similar sensitivity to climate change. Their positive responses to precipitation in previous December consistently increased, as well as the negative correlation with precipitation in current September. As to P. masso-niana, they had a relatively stronger climatic sensitivity and higher stability than the other two species. It would be more suitable for P. massoniana trees on the southern slope of the Funiu Mountains under global warming.
Assuntos
Pinus , Traqueófitas , Mudança Climática , Árvores , China , Aquecimento GlobalRESUMO
OBJECTIVE: To explore the effect of warm-needle moxibustion (WNM) on the levels of T cell subgroups and serum inflammatory factors, intestinal microecological balance and postoperative adverse reactions in patients with colorectal cancer. METHODS: Eighty-four patients who underwent elective radical resection of colorectal cancer were randomly and equally divided into control (medication) group (23 men and 19 women) and WNM group (24 men and 18 women). Patients of the control group received conventional medication treatment (such as postoperative anti-infection and fluid supplementation), and those of the WNM group received conventional medication plus WNM stimulation (the acupuncture needle handle warmed by ignited moxa stick) of bilateral Zusanli(ST36), Sanyinjiao(SP6), Yinlingquan(SP9), Shangjuxu(ST37), and Zhaohai(KI6). The acupuncture needles were retained for 45 minutes every time, starting on the first day after surgery, once a day for 15 days. The number of T cell subsets (CD3+, CD4+, CD8+) positive cells was counted under fluorescence microscope, and the contents of serum tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6) were detected by using ELISA, and the level of C-reactive protein (CRP) was detected by using immunoturbidimetry. The levels (logarithm of colony-forming units per gram of wet fecal weight) of Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus were determined. The adverse reactions (leukocyte decline, nausea and vomiting, peripheral phlebitis, cold stimulation sensitivity) were recorded after surgery. RESULTS: Before treatment, there were no significant differences between the two groups in the number of T cell subgroups, TNF-α and IL-6 contents, and intestinal flora numbers (P>0.05). After the treatment, self-comparison showed that the numbers of CD3+ and CD4+ positive cells, the ratio of CD4+/CD8+ and the intestinal Bifidobacterium and Lactobacillus levels in the WNM group were significantly increased (P<0.05), whereas the number of CD8+positive cells, intestinal Escherichia coli and Enterococcus levels in the WNM group, and the levels of TNF-α, IL-6 and CRP in both groups were obviously decreased in comparison with their own pretreatment (P<0.05), but no significant changes were found in the levels of CD3+ and CD4+ positive cells, CD4+/CD8+ and intestinal Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus in the control group (P>0.05). Comparison between two groups displayed that after the treatment, the numbers of CD3+ and CD4+ positive cells, the ratio of CD4+/CD8+, as well as the levels of Bifidobacterium and Lactobacillus were significantly higher in the WNM group than in the control group (P<0.05), whereas the number of CD8+ positive cellsï¼ TNF-a, IL-6 and CRP, and the levels of Escherichia coli and Enterococcus were obviously lower in the WNM group than in the control group (P<0.05). The incidence of adverse reactions including leukopenia, nausea and vomiting, peripheral phlebitis, and sensitivity to cold stimulation in the WNM group were markedly lower than those of the control group (P<0.05). CONCLUSION: WNM intervention can significantly improve the immune function, reduce the level of inflammatory factors, regulate the level of beneficial intestinal flora, and also reduce the incidence of postoperative adverse reactions in patients experiencing radical resection of colorectal cancer.
Assuntos
Neoplasias Colorretais , Microbioma Gastrointestinal , Moxibustão , Neoplasias Colorretais/cirurgia , Feminino , Humanos , Imunidade , Masculino , AgulhasRESUMO
The methods of fuzzy cluster and curve-fitting combined with FTIR were used to determine the origins of Herba Abri cantoniensis and Herba Abri mollis. The spectra of Herba Abri cantoniensis and Herba Abri mollis are similar, both with typical spectral shapes. The two spectra can be divided into 3 parts: the 1st is 3 500-2 800 cm(-1), containing stretching bands of -OH, N-H, and CH2 ; the 2nd is 1 800-800 cm(-1), containing stretching bands of ester carbonyl group and indican C-O(H), vibrational bands of C=C and benzene ring; The 3rd is 800-400 cm(-1), containing skeletal vibration and scissoring vibration of molecular. The recorded FTIR spectral data were processed by 9-point-smoothing, 1st derivative, SNV and fuzzy cluster analysis sequentially. The fuzzy cluster analysis was carried out by similarity or dissimilarity matrix, and two matrices are computed with Manhattan and Euclidean distance. The results indicated that the optimization used Manhattan and dissimilarity matrix, and 5 origins of Herba Abri cantoniensis were perfectly discriminated, but 2 origins of Herba Abri mollis were mixed and identified from the other 3 origins. So the characterized bands at 1 034 cm(-1) of the average 1-D spectra of Herba Abri cantoniensis and Herba Abri mollis were fitted combining 2nd derivative for further distinguishing their spectral characteristic. The results of curve-fitting showed that the bands of wild Herba Abri cantoniensis and the other origin ones were decomposed to 11 and 9 component bands respectively, but the bands of Shanglin and the other origins Herba Abri mollis were decomposed to 9 and 8 component bands dissimilarly, and the locations and normalized densities of these component bands were different. From this, together with the results of fuzzy cluster analysis, it is concluded that the combination of two methods may identify the origins of Herba Abri cantoniensis and Herba Abri mollis availably.
Assuntos
Abrus/química , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The methods of sequential analysis of dual-indexes and cluster analysis were utilized to investigate the infrared fingerprints of A. cantoniensis planted in different years and different places in Guangxi, China. The results showed that 6 samples were able to be completely separated only through 13 point smoothing, when the dual-indexes analysis was applied in the present research, and the accurate relationship between these samples could be inspected and expressed by quantitative relationships under 6-dimensional spaces; however, the effect of cluster was bad only through 13 point smoothing of raw spectra, and it was very difficult to find out the regular sequences while the cluster analysis was applied. Furthermore, the 6 samples were able to be completely separated if raw spectra were dealed with 1st derivative after 13 point smoothing, and the clustering effects were more obvious and 6 samples of A. cantoniensis were completely separated. The above two methods could be used to evaluate the quality of Chinese medicinal materials easily when the sample was not excessive quantitatively, but the method of dual-indexes analysis was more difficult than the clustering analysis if the sample size was too large, since a mass of data such as common peak ratio and variation peak ratio of the IR fingerprint spectra were processed and analyzed statistically, while this method could accurately find out the closest relationship between any samples through comparing the quantitative relationships of common peak ratio and variation peak ratio of each sample under 6-dimensional space; the precision of cluster analysis was less than dual-indexes analysis, but it was more convenient than dual-indexes analysis when large sample data were analysed. Finally the above two methods all showed that the chemical composition of the A. cantoniensis was similar in the same cultivated area, but the difference in chemical composition of A. cantoniensis in different years was distinct even they were in the same place.
Assuntos
Abrus/química , Medicamentos de Ervas Chinesas/química , Espectrofotometria Infravermelho , China , Análise por Conglomerados , Geografia , Fatores de TempoRESUMO
Paeonol is a simple phenolic compound isolated from herbal root bark, which has been reported to possess numerous biological and pharmacological characteristics, including a desirable antitumor effect. To date, the effect of paeonol against colorectal cancer (CRC) cells is yet to be fully elucidated. Therefore, the present study aimed to identify the underlying mechanism via which paeonol exerts its antitumor activity on HCT116 cells. After incubation with various concentrations of paeonol (7.8125, 15.625, 31.25, 62.5, 125, 250 and 500 µg/ml), the inhibitory effect of paeonol on cell viability was assessed using a Cell Counting Kit8 assay. Cell apoptosis and cell cycle distribution were measured using flow cytometry. Moreover, caspase activity was measured using a colorimetric caspase assay. Luciferase assay was also used to determine the ßcateninmediated transcriptional activity of Tcell specific transcription factor/lymphoidenhancer binding factor (TCF/LEF), and western blotting analysis was performed to measure the related expression of proteins. The results indicated that paeonol exhibited a notable effect against HCT116 cells by inducing G0/G1phase arrest, as demonstrated by downregulation of the cell cycle regulators cyclindependent kinase 4 and cyclin D1 and upregulation of p21Cip1 in a dosedependent manner. Furthermore, paeonol dosedependently induced cell apoptosis, accompanied by an increase in the Bax/Bcl2 ratio, release of cytochrome c and further activation of caspases. Paeonol also dosedependently blocked the activation of the Wnt/ßcatenin signaling pathway by suppressing the expression of ßcatenin, resulting in a decrease in ßcateninmediated activity of TCF/LEF and downregulation of downstream target genes, including cyclin D1, survivin and cMyc. Therefore, the present results suggested that paeonol exerted its antitumor effects on CRC cells, including the inhibition of cell proliferation, induction of cell cycle arrest and initiation of apoptosis, at least partly by suppressing the Wnt/ßcatenin pathway, which may offer a promising therapeutic strategy for CRC.
Assuntos
Acetofenonas/farmacologia , Neoplasias Colorretais/metabolismo , Apoptose/efeitos dos fármacos , Western Blotting , Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ciclina D1/metabolismo , Quinase 4 Dependente de Ciclina/metabolismo , Fase G1/efeitos dos fármacos , Células HCT116 , Humanos , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Survivina/metabolismo , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
Our objective was to explore the mechanism of spectral reflectance response to leaf photosynthetic pigment seasonal dynamic of Quercus aliena var. acuteserrata under throughfall elimination. We simulated rainfall decline through throughfall elimination (TFE) treatment in the experiment plots of Baotianman Natural Reserve in Henan, measured leaf pigment content and its spectral reflectance during growing season in both TFE and control plots. We analyzed seasonal changes of photosynthetic pigments and changes of pigments induced by TFE and their spectral reflectance responses. The results showed that all photosynthetic pigments content and pigment-ratios exhibited clear seasonal patterns. Leaf photosynthetic pigments content and ratio had differences between TFE plot and control plot during the whole growing season, and significant difference was found in chlorophyll b (Chl b) indicating that Chl b had higher sensitivity than other pigments. Carotenoids (Car) content showed minor difference compared with other pigments, indicating that Car had less sensitivity to TFE. The spectral reflectance of 550 nm was found to be the waveband most sensitive to seasonal changes of pigments, so we formulated the sample ratio index (SR750,550) based on it. The strong positive relationships between SR750,550 and Chl a, Chl b, Chl and Car contents were found with high significant level. Similarly, significant negative relationships were also been found between photochemical reflectance index (PRI) and Car/Chl. The spectral reflectance of 550 nm was most sensitive to changes of pigments induced by TFE. SR750,550 was sensitive to changes of leaf Chl a, Chl b and Chl content induced by TFE (Pï¼0.01), but not to change of Chl a/b. PRI was sensitive to change of leaf Car/Chl induced by TFE (Pï¼0.01).
Assuntos
Fotossíntese , Quercus , Carotenoides , Clorofila , Folhas de Planta , Estações do AnoRESUMO
OBJECTIVE: To study the clinical significance and reliability of strand displacement amplification (SDA) in detecting Mycobacterium tuberculosis complex. METHODS: SDA and fluorescence quantitative polymerase chain reaction (FQ-PCR) were employed to detect samples from 453 cases of tuberculosis, including 332 sputum samples, 78 samples of pleural effusion, and 43 samples of cerebrospinal fluid. RESULTS: In the 332 sputum samples, 131 were culture-positive, of which 110 samples (88 smear-positive) were Mycobacterium tuberculosis complex and 21 samples (20 smear-positive) were nontuberculous Mycobacteria. The sensitivity and specificity of SDA for the 110 samples of Mycobacterium tuberculosis complex and 21 samples of nontuberculous Mycobacteria were 99.1%, 95.2% and 94.6%, 95.2%, respectively. The positive rates in the 311 cases of Mycobacterium tuberculosis complex for SDA and FQ-PCR were 55.3% (172/311) and 47.0% (146/311), respectively. There were 20 smear positive samples in the 121 samples of pleural effusion and cerebrospinal fluid, of which 19 were Mycobacterium tuberculosis, 1 nontuberculous Mycobacterium. The positive rates for SDA and FQ-PCR were 43.4% (52/120) and 33.4% (40/120), respectively. Internal amplification control (IAC) was designed for SDA to achieve accuracy of the results. CONCLUSION: The automatic assay system of SDA is a rapid and specific test for detecting Mycobacterium tuberculosis complex.
Assuntos
Técnicas Bacteriológicas/métodos , Mycobacterium tuberculosis/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Tuberculose/diagnóstico , DNA Bacteriano , Humanos , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Escarro/microbiologia , Tuberculose/microbiologiaRESUMO
sDC-SIGN is the soluble form of dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN, CD209), which is a molecule involved with pathogen recognition and immune regulation. However, there is no commercially available ELISA kit for detecting human sDC-SIGN, and the normal range of this molecule is unknown. Here, we describe an ELISA for detecting human sDC-SIGN with high specificity. First, sDC-SIGN protein was expressed and purified. Monoclonal and polyclonal antibodies were then raised against the purified protein and subsequently characterized. A sandwich ELISA was developed using polyclonal antibodies specific for sDC-SIGN for capture and a biotin-labeled monoclonal antibody specific for sDC-SIGN for detection of protein. This method has sensitivity up to 0.2 ng/ml. Using this ELISA, we found that the concentration of sDC-SIGN in sera of healthy volunteers ranges from 0-319 ng/ml with a mean concentration of 27.14 ng/ml. Interestingly, the concentration of sDC-SIGN in sera from patients with cancer or chronic hepatitis B virus (CHB) infection was lower than that of health controls. The mean concentrations of sDC-SIGN in cancer patients and chronic hepatitis B virus infection patients were 3.2 ng/ml and 3.8 ng/ml, respectively. We developed a sandwich ELISA for detecting human sDC-SIGN and demonstrated its use by assessing sera concentrations of sDC-SIGN in patients with cancer and chronic CHB infection compared to that of healthy controls.
Assuntos
Moléculas de Adesão Celular/isolamento & purificação , Células Dendríticas/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Hepatite B Crônica/sangue , Lectinas Tipo C/isolamento & purificação , Neoplasias/sangue , Receptores de Superfície Celular/isolamento & purificação , Animais , Anticorpos Monoclonais/imunologia , Estudos de Casos e Controles , Moléculas de Adesão Celular/sangue , Feminino , Hepatite B Crônica/diagnóstico , Humanos , Lectinas Tipo C/sangue , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/diagnóstico , Coelhos , Receptores de Superfície Celular/sangue , Sensibilidade e EspecificidadeRESUMO
AIM: To generate and identify monoclonal antibodies (mAb) against homo sapiens hydroxysteroid 11-beta dehydrogenase 1 (HSD11B1). METHODS: Normal human liver tissues were homogenized, and mitochondria were isolated by differential centrifugation. The total mitochondrial proteins were used to immunize BALB/c mice to generate mAb by hybridoma technique. The mAb were characterized by ELISA, Western blot and immunohistochemistry. The antibody specificity was identified by immunoprecipitation (IP), and confirmed by Uni-ZAP expression library screening. RESULTS: One hybridoma CBF245 secreting specific mAb against HSD11B1 was established. The Ig subclass of this mAb was IgG1, and it could be used in ELISA, Western blot, immunohistochemistry. Our data showed that the antigen recognized by CBF245 mAb was localized in the hepatocyte cytoplasm, with molecular weight of M(r) 35 000 in the mitochondria of human liver tissue. The CBF245 mAb was further confirmed by immunoscreening of Uni-ZAP XR liver cDNA expression library. CONCLUSION: A hybridoma cell line stably secreting specific mAb against HSD11B1 is established and characterized. This mAb reacted with HSD11B1 in ELISA, Western blot, immunohistochemistry assay, IP, and would be very useful for the HSD11B1 studies.
Assuntos
Anticorpos Monoclonais/biossíntese , Especificidade de Anticorpos , Hibridomas/imunologia , 11-beta-Hidroxiesteroide Desidrogenase Tipo 1/imunologia , Animais , Anticorpos Monoclonais/imunologia , Humanos , Imunoglobulina M/imunologia , Imuno-Histoquímica , CamundongosRESUMO
AIM: To generate monoclonal antibodies (mAb) against glyoxylate reductase/hydroxypyruvate reductase (GRHPR). METHODS: Normal human liver tissues were homogenized, and human liver cytosolic proteins were isolated by centrifugation. The total human liver cytosolic proteins were used to immunize BALB/c mice to prepare mAb by hybridoma technique. The mAbs were detected by ELISA, Western blot, and immunohistochemistry. The antibody specificities were identified by mass spectrometry (MS) following immunoprecipitation (IP) and confirmed by Uni-ZAP expression library screening. RESULTS: One hybridoma cell line, ADB291, secreting specific mAb against GRHPR was established. The Ig subclass of the mAb was IgG1(kappa). Data from immunohistochemistry showed that ADB291 can recognize hepatocyte cytoplasm. ADB291 mAb was used to isolate its protein antigen by IP. Proteins captured by the mAb were loaded to SDS-PAGE and subjected to Western blot and MALDI-TOF MS analysis. lambda expression Uni-ZAP XR pre-made liver cDNA library was screened with ADB291 hybridoma supernatants. All of our data demonstrated that ADB291 mAb specially reacted with GRHPR. CONCLUSION: A hybridoma cell line stably secreting specific mAb against GRHPR is established. The specific mAb against GRHPR would be very useful for the studies of GRHPR functions and distribution.
Assuntos
Oxirredutases do Álcool/imunologia , Anticorpos Monoclonais/imunologia , Animais , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/metabolismo , Especificidade de Anticorpos , Western Blotting , Linhagem Celular , Citosol/metabolismo , Eletroforese em Gel de Poliacrilamida , Biblioteca Gênica , Hepatócitos/citologia , Humanos , Hibridomas/metabolismo , CamundongosRESUMO
This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.
Assuntos
Anticorpos Monoclonais/biossíntese , Hemoglobina A2/imunologia , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Sequência de Bases , Humanos , Hibridomas/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Talassemia alfa/imunologiaRESUMO
The study was aimed to generate monoclonal antibodies (mAbs) against homo sapiens UDP-glucose pyrophosphorylase 2 (UGP2). Normal human liver tissues homogenized, and cytosolic proteins isolated by centrifugation were used to immunize BALB/c mice to generate mAbs by hybridoma technique. The mAbs were identified by ELISA, Western blot, and immunohistochemistry assay. The antibody specificity was confirmed by Uni-ZAP expression library screening. The results indicated that one hybridoma BAD062 secreting specific mAb against UGP2 was established. The Ig subclass of this mAb was IgG(2b) (kappa), and it could be used in ELISA, Western blot, immunohistochemistry assay. The antigen recognized by BAD062 mAb was localized in the hepatocyte cytoplasm, with molecular weight of 56 kD in the cytosolic proteins of human liver tissue. The BAD062 mAb was further confirmed by immunoscreening of Uni-ZAP XR liver cDNA expression library. It is concluded that a hybridoma cell line stably secretes specific mAb against UGP2. This mAb reacted with UGP2 in ELISA, Western blot, immunohistochemistry assay, and would be very useful for the UGP2 studies.
Assuntos
Anticorpos Monoclonais/biossíntese , UTP-Glucose-1-Fosfato Uridililtransferase/imunologia , Animais , Anticorpos Monoclonais/análise , Especificidade de Anticorpos , Sequência de Bases , Humanos , Hibridomas/metabolismo , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência MolecularRESUMO
OBJECTIVE: To evaluate of therapeutic efficacy of deoxyribouncleotidum on pulmonary tuberculosis. METHODS: Eighty patients with pulmonary tuberculosis sustaining hepatic lesion after treatment with antituberculosis drugs were randomized into therapeutic group and control group. Patients in the control group received regular treatment and those in the therapeutic group had additional deoxyribouncleotidum injection. RESULTS: ALT, AST, ALP and TBIL levels were significantly higher in the therapeutic group than in the control group 4 weeks after treatment. IgG, IgA, IgM levels, and CD3(+) and CD8(+) lymphocytes were significantly increased in the therapeutic group after treatment (P<0.05). CONCLUSION: deoxyribouncleotidum can improve hepatic function and immunity in patients with pulmonary tuberculosis.