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Body mass index (BMI) has been increasing globally in recent decades. Previous studies reported that BMI was associated with sex hormone levels, but the results were generated via linear regression or logistic regression, which would lose part of information. Quantile regression analysis can maximize the use of variable information. Our study compared the associations among different regression models. The participants were recruited from the Center of Reproductive Medicine, The First Hospital of Jilin University (Changchun, China) between June 2018 and June 2019. We used linear, logistic, and quantile regression models to calculate the associations between sex hormone levels and BMI. In total, 448 men were included in this study. The average BMI was 25.7 (standard deviation [s.d.]: 3.7) kg m-2; 29.7% (n = 133) of the participants were normal weight, 45.3% (n = 203) of the participants were overweight, and 23.4% (n = 105) of the participants were obese. The levels of testosterone and estradiol significantly differed among BMI groups (all P < 0.05). In linear regression and logistic regression, BMI was associated with testosterone and estradiol levels (both P < 0.05). In quantile regression, BMI was negatively associated with testosterone levels in all quantiles after adjustment for age (all P < 0.05). BMI was positively associated with estradiol levels in most quantiles (≤80th) after adjustment for age (all P < 0.05). Our study suggested that BMI was one of the influencing factors of testosterone and estradiol. Of note, the quantile regression showed that BMI was associated with estradiol only up to the 80th percentile of estradiol.
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Male , Humans , Body Mass Index , Cross-Sectional Studies , Gonadal Steroid Hormones , Regression Analysis , Estradiol , TestosteroneABSTRACT
BACKGROUND@#Solitary pulmonary nodule has received increasing attention in recent years. A couple of lung nodules have been recognized as primary malignant tumors, which leads to an urgent need in enhancing the diagnosis of benign/malignant lung nodules at clinical settings. This study aims to explore the value of the combined detection of cytokines and tumor markers in differencing benign and malignant solitary pulmonary nodules in diagnose.@*METHODS@#With 81 solitary pulmonary nodules cases with a clear diagnosis, the general clinical data, nodule imaging features, pathological diagnosis data, serological index cytokine series and tumor marker expression levels were collected in groups. Both single factor and multi-factors analysis were conducted to screen out the serum influence indexes that can predict the malignant probability of lung nodules, and mean while binary logistic regression analysis was used to construct joint indexes; After receiver operating characteristic curve (ROC) was drawn, the area under the curve and the corresponding sensitivity, specificity and positive of each index predicted value, negative predicted value and accuracy could be calculated with a view to determine the statistical significance of area under the curve (AUC).@*RESULTS@#There are differences in the distribution of malignant solitary pulmonary nodules at different locations, with the highest proportion of the right upper lobe (40.4%). The serum levels of carcinoembryonic antigen (CEA), cytokeratin 19 fragment 21-1 (CYFRA21-1), interleukin-6 (IL-6), interleukin-8 (IL-8) in the malignant nodule group were higher than those in the benign nodule group. Logistic regression analysis suggests that CEA, IL-6 and IL-8 are independent risk factors for predicting malignant nodules. ROC curve analysis shows that the areas under the curve of the individual indicators CEA, IL-6 and IL-8 are 0.642, 0.684 and 0.749. The comparison result of the test efficiency of the area under the curve suggests that CEA+IL-6+IL-8 has a larger area under the curve and higher detection efficiency.@*CONCLUSIONS@#CEA, IL-6 and IL-8 are independent risk factors for malignant solitary pulmonary nodules. The combined detection of cytokines and tumor markers has played a role in the differential diagnosis of benign and malignant lung nodules. The diagnostic value of the combined detection of CEA+IL-6+IL-8 is the highest.
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A 56-year-old female patient was admitted to a hospital because of repeated fatigue, anorexia, fever for more than one year, recurrence for 2 months and sudden left limb twitch for 10 days. In 2017, patient was diagnosed with infective endocarditis, then underwent aortic valve replacement, valve biopsy showed Candida parapsilosis, she took medicine regularly after operation and withdrew medicine after three consecutive negative blood culture. Fever and multiple organ infarction occurred again in 2018, Candida parapsilosis was isolated again from blood culture, it is suggested that fungal endocarditis may be latent and recurrence in human body for a long time, exfoliation of vegetation may embolize various organs and lead to corresponding clinical manifestations, surgery combined with drug therapy and subsequent maintenance of antifungal therapy are extremely important to improve the prognosis of patients.
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Objective To establish and evaluate a rat model of inhalation lung injury induced by ship smog.Methods A rat model of inhalation lung injury was established by analyzing the composition of ship materials after combustion.Fortytwo healthy male Wistar rats were randomly divided into normal control group and 2,6,12,24,48 and 72h groups (6 each)after inhalation,these rats were killed at each time point,and the changes of arterial blood gas,coagulation function,the lung water content (％) were detected.Macroscopic and microscopic changes in lung tissues were observed to judge the degree of lung injury.Results The main components after combustion of 7 kinds of nonmetal materials on ship included CO,CO2,H2S,NOx and other harmful gases in this study,AIKE in one gas detector was used to monitor O2,CO,CO2 and H2S,and their concentrations remained relatively stable within 15 minutes,and the injury time was 15 minutes.The rats presented with shortness of breath and mouth breathing.Smoke inhalation caused a significant hypoxemia,the concentration of blood COHb reached a peak value 2h and the lung water content (％) did 6h after inhalation (P＜0.05).It is metabolic acidosis in the early stage after inhalation,but metabolic acidosis combined with respiratory acidosis in the later period.Histopathological observation showed diffuse hemorrhage,edema and inflammatory cell infiltration in the lung tissue as manifestations of lung injury,and the injury did not recover at 72h after inhalation,the change of blood coagulation function was not statistically significant.Conclusion A rat model of inhalation lung injury induced by ship smog has been successfully established,and has the advantages of easy replication,stability and reliability,thus can be used to research and treat inhalation lung injury induced by ship smog in naval war environment and other cases.
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Objective To investigate the effect and safety of Gukang capsule combined with sodium hyaluronate injection on the treatment of the patients with osteoarthritis. Methods 94 osteoarthritis patients selected from December 2015 to March 2017 were randomly divided into study group (n=47) and control group (n=47). The control group were given sodium hyaluronate injection, the study group were received local injection of sodium hyaluronate combined with Gukang capsule. The changes of knee joint function (HSS scale score) and adverse reaction rate were recorded and compared before and after treatment in the two groups. Results Before treatment, there was no significant difference in HSS scores between the two groups. After treatment, the HSS score in the study group was significantly better than that in the control group (P<0.05). There were no significant differences in the adverse reactions between the 2 groups during the treatment. Conclusion Local injection of sodium hyaluronate combined with Gukang capsule on the treatment of osteoarthritis, which can significantly improve the clinical efficacy, the security is higher. .
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Objective To investigate the effect and safety of Gukang capsule combined with sodium hyaluronate injection on the treatment of the patients with osteoarthritis. Methods 94 osteoarthritis patients selected from December 2015 to March 2017 were randomly divided into study group (n=47) and control group (n=47). The control group were given sodium hyaluronate injection, the study group were received local injection of sodium hyaluronate combined with Gukang capsule. The changes of knee joint function (HSS scale score) and adverse reaction rate were recorded and compared before and after treatment in the two groups. Results Before treatment, there was no significant difference in HSS scores between the two groups. After treatment, the HSS score in the study group was significantly better than that in the control group (P<0.05). There were no significant differences in the adverse reactions between the 2 groups during the treatment. Conclusion Local injection of sodium hyaluronate combined with Gukang capsule on the treatment of osteoarthritis, which can significantly improve the clinical efficacy, the security is higher. .
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Objective To study the performance indicators after reorganization for 10 Years in enterprise hospital.Methods The performance indices from 2005 to 2014 of a Class One Grade A hospital was evaluated by rank sum ratio.Results The reform was in the doldrums in 2005, improved slightly in 2006 and 2007, increased significantly and steadily in 2008, reached the best in 2014.Conclusion Evaluating the performance of hospital management with rank sum ratio is scientific and clear, which has certain practical value.
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Professor WEI Pin-kang believes that, as a complication of gastric cancer, malignant ascites and gastric cancer have the same pathogenesis: water and dampness retention is the external performance and phlegm resistance is the inner essence. Hence therapy of resolving phlegm and promoting diuresis is suggested for the fundamental treatment of gastric ascites. Using the method of the combination of internal and external treatment, this therapy includes resolving phlegm and draining water medicine both to dispel pathogen and to improve symptoms. Xiaotan LishuiDecoction based on this therapy is composed of Arisacma Cum Bile, processed Pinelliae Rhizome, Cremastrae Pseudobulbus Pleiones Pseudobulbus, old pericarp of bottle gourd, Polyporus, Natriee Sulfas, Phytolaccae Radix and Poria.
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AIM: To compare intraocular pressure (IOP) fluctuations measured at home and in the clinic over a 24-hour period.METHODS: A prospective investigational study.A total of 120 Chinese participants were selected from five communities in the Chengdu area.Patients underwent a clinical interview and IOP was measured both at home and in the clinic.IOP were measured at 8 a.m., 10 a.m., 12 a.m., 2 p.m., 4 p.m., 6 p.m., 8 p.m., 10 p.m., 2 a.m., 6 a.m.using the same pneumatonometer.Measurements were taken in the sitting position.RESULTS: The average 24-hour IOP measured in the clinic was slightly lower than that at home.The mean difference in 24-hour IOP measurements between home and clinic was 0.27 mmHg.The IOP fluctuation in the clinic was higher than at home (the mean difference was 0.01 mmHg).There was no statistically significant difference in the average 24-hour IOP measured at home vs in the clinic.The average IOP measured at 2 p.m.at home (16.04±5.95 mmHg) was significantly higher compared with the measurement in the clinic (15.43±5.16 mmHg) (P<0.05).The overall agreement between 24-hour IOP measurements made in the clinic and at home in diagnosis of primary open angle glaucoma was 85.0% (K coefficient: 0.68).CONCLUSION: The 24-hour IOP measured in the clinic was similar to that measured at home, and the method of measuring IOP in the clinic is acceptable in diagnosing primary open angle glaucoma.
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<p><b>OBJECTIVE</b>To study the effect of Flos Daturae alkaloids (FDA) on TGF-beta1-1uuuu;U epithelial-mesenchymal transition (EMT) of human pulmonary adenocarcinoma A549 cells.</p><p><b>METHODS</b>A549 cells in vitro cultured were randomly divided into 5 groups, i.e., the blank control group, the TGF-beta1 group, the low dose FDA group, the medium dose FDA group, and the high dose FDA group. The morphologies of A549 cells were observed. Expressions of cytokeratin (CK)-19 and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot and real-time PCR at 24, 48, and 72 h, respectively.</p><p><b>RESULTS</b>A549 cells in the TGF-beta1, group turned from cobblestone to spindle shape gradually. Those in low, medium and high dose FDA groups showed similar shapes to those of the TGF-beta1 group. There was no statistical difference in the morphology of A549 cells among the 3 dose FDA groups (P > 0.05). Western blot showed that, when compared with the blank control group, the expression of CK-19 was down-regulated, but the expression of alpha-SMA was up-regulated in the TGF-beta1 group (P < 0.01). Compared with the TGF-beta1, group, the expression of CK-19 was up-regulated, but the expression of alpha-SMA was suppressed in low, medium and high dose FDA groups (P < 0.01). The CK-19 expression obviously increased, but the alpha-SMA expression was suppressed in high dose FDA group at 72 h (P < 0.01). Real-time PCR results showed, as compared with the TGF-beta1 group, the mRNA expression of CK-19 was increased, but the mRNA expression of alpha-SMA was reduced in low, medium and high dose FDA groups (P < 0.01).</p><p><b>CONCLUSIONS</b>FDA had no effect on EMT morphological changes of TGF-beta1 induced A549 cells. FDA could reverse characteristic markers of A549 cells during EMT to some extent, such as expressions of CK-19 and alpha-SMA. The expression of CK-19 (as the epithelium marker) increased and the expression of alpha-SMA (as the mesenchymal marker) was reduced. Besides, they were most obviously seen in the high dose FDA group at 72 h in a dose- and time-dependent manner.</p>
Subject(s)
Humans , Actins , Adenocarcinoma , Alkaloids , Pharmacology , Cell Line, Tumor , Datura , Drugs, Chinese Herbal , Pharmacology , Epithelial Cells , Epithelial-Mesenchymal Transition , Epithelium , Transforming Growth Factor beta1 , MetabolismABSTRACT
Objective To investigate the effects of microRNA-155 (miR-155) inhibitor on JAK/STAT1 (Janus kinase/signal transducer and activator transcription 1) signaling pathways in the injured lung tissue induced by lipopolysaccharide (LPS).Methods One hundred and twenty BALB/c mice were randomly divided into control group (n =40),LPS group (n =40),and inhibitor + LPS group (n =40).LPS group and inhibitor + LPS group were made by injection of LPS 20 mg/kg intra-peritonealy,whereas equivalent volume of normal saline was given instead in the control group.The 80 mg/kg of miR-155 inhibitor was injected into caudal vein 24 h before LPS injection in inhibitor + LPS group.Mice were sacrificed at 6 h,12 h,24 h,and 48h separately after LPS injection,and lung tissue were collected.The levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) of lung tissue were measured using the enzyme-linked immunosorbent assay (ELISA).Using histopathological examination,the injury of lung tissue was evaluated.The expressions of miR-155,STAT1 mRNA,SOCS1 mRNA in lung tissue were assayed by fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results The miR-155 expression induced by LPS increased at 6 h,12 h,24 h and decreased at 48 h.The miR-155 expressions in LPS group were (8.52 ± 1.12) at 6 h,(11.04 ±0.99) at 12 h,(15.84 ±0.80) at 24 h and (4.03 ± 2.55) at 48 h.In the inhibitor + LPS group,the expressions of miR-155 were lower compared with LPS group,showing significant differences at 12 h (t =6.08,P ＜ 0.01),and at 24 h (t =23.64,P ＜ 0.01).STAT1 mRNA and SOCS1 mRNA both reached peak levels at 6 h after LPS injection.The levels of STAT1 mRNA in LPS group were higher than those in inhibitor + LPS group,showing significant differencesat6h (t=4.41,P＜0.01),12h(t=2.69,P＜0.05),and24h (t=3.62,P＜0.01).The levels of SOCS1 mRNA in inhibitor + LPS group were higher than those in LPS group,showing significant differences at 6 h (t =4.55,P ＜0.01),12 h (t =4.12,P ＜0.01),24 h (t =2.38,P ＜ 0.05).TNF-α reached its peak value at 6 hours and IL-10 reached its peak value at 48 hours.Both TNF-α and IL-10 were higher in LPS group than those in inhibitor + LPS group showing significant differences at 6 h,12 h,24 h (P ＜0.01).The pathologic examination indicated the lung injury in inhibitor + LPS group was milder than that in LPS group.Conclusion The miR-155 increased in lung tissue of endotoxemic mice.miR-155 inhibitor may suppress JAK/STAT1 signaling pathway and protect the lung tissue.
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Objective To observe inhibitory effect of recombinant human endostatin on the proliferation of vascular tumor endothelial cells and to investigate its possible mechanism.Methods Hemangioma endothelial cells were cultured in vitro and different concentrations of endostatin on hemangioma endothelial cell proliferation inhibition were detected by MTT method.Effect of recombinant human endostatin on endothelial cell cycle was detected by flow cytometry.The expression of VEGF, KDR mRNA in hemangioma endothelial cell were detected by Real-time RT PCR. Results Recombinant human endostatin concentrations in 24 h, 48 h and 72 h after three period during which the hemangioma endothelial cells inhibited significantly( P <0.01 ) , and there was a clear dose dependence, IC50 was 355 μg/mL.Recombinant human vascular endostatin ( 250 μg/mL ) intervented for 24 hours, the proportion of cells in G0/G1 phase(94.23 ±1.66)%, compared with control group (90.63 ±1.14)%, had significantly difference (P<0.05).Compared with the control group, the difference of the expression of vascular endothelial growth factor (VEGF) was statistically significant (P<0.05) as well as Flk-1 (P <0.05).Conclusion Recombinant human endostatin does not only have inhibitory effect of hemangioma endothelial cell cycle, but also can inhibit the expression of VEGF and FLK-1.
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This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Subject(s)
Humans , Amino Acid Motifs , Influenza A Virus, H1N1 Subtype , Chemistry , Genetics , Virulence , Influenza A Virus, H5N1 Subtype , Chemistry , Genetics , Virulence , Influenza, Human , Virology , Neuraminidase , Chemistry , Genetics , Metabolism , Viral Proteins , Chemistry , Genetics , Metabolism , VirulenceABSTRACT
Objective To investigate the characteristic changes of the metabolism products in auditory cortex (transverse temporal gyrus) in patients with OSAHS combined with nerve deafness using 1 H magnetic resonance spectroscopy (1 H - MRS) ,and to discovery the early warning indicator of nerve deafness in OSAHS .Methods PTA was performed in 95 patients with OSAHS (diagnosed by PSG) ,and 15 healthy control subjects ,the patients were classified into four groups :the group of OSAHS ;OSAHS with unilateral and bilateral deafness ,the normal control group .Cerebral metabolism was studied by assessing the ratios of nitro -acetyl aspartate contrast to choline (NAA/Cho) as well as to creatine (NAA/Cr) ,myo -inositol to creatine (mI/Cr) and choline to creatine (Cho/Cr) ratios in the auditory cortical separately in these groups .ROC curves were made for those metabolism markers to find the best diagnositic threshold .Results Significantly lower values of NAA/Cho ratio were found in patients'(OSAHS with deafness) auditory cortex compared with 15 age-matched control subjects (P<0 .05) and OSAHS without deafness (P<0 .05) .Auditory cortical NAA and NAA/Cho ratio in OSAHS with unilateral nerve sensorineual hearing were significantly lower than those of in normal control (P<0 .05) ,but there was no significant difference be-tween the abnormal and ontralateral normal auditory cortex by a self comparison .All of the metabolisms were tested by the curve of ROC .Conclusion Combined with the changes of metabolism ,and the curve of ROC ,NAA/Cho may be the early warning markers of sensorineual hearing was in OSAHS patients .
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To describe the unique miRNA profiles for HIV seropositive individuals and identify significantly differently expressed miRNAs, we determined the expression level of 754 miRNAs of 10 HIV seropositive individuals and 10 HIV seronegative individuals by using the Taqman low density microRNA array. BRB-Array Tool was used to conduct the significance analysis, and the DIANA online tool was used to perform the miRNA target prediction and pathway analysis. A total of 56 significantly differentially expressed miRNAs were identified by microarray between HIV seropositive and seronegative individuals. Among them, 49 miRNAs were down-regulated and 7 were up-regulated, partially overlapped with reported data. Predicted target genes were mainly involved in MAPK, TGF-beta and Wnt pathways. The results shows that miRNA profile changes in HIV-1 seropositive individuals, and the 56 differentially expressed miRNAs may play important role during HIV infection. Further studies on these miRNAs may be helpful for identify key molecules involved in HIV infection and potential diagnostic markers.
Subject(s)
Adult , Female , Humans , Male , Young Adult , Gene Expression Profiling , HIV Infections , Blood , Genetics , Virology , HIV-1 , Genetics , Physiology , MicroRNAs , Blood , GeneticsABSTRACT
OBJECTIVE@#In this study, we investigated the anti-inflammation effects of Xuebijing in OVA-induced murine allergic rhinitis model. Furthermore, we determined whether heme oxygenase (HO)-1 is required for the protective activity of Xuebijing.@*METHOD@#Airways of OVA-sensitized mice exposed to OVA challenge developed eosinophilia, mucus hypersecretion and increased cytokine levels. Levels of interleukin IL-4, IL-5, IL-13, and tumor necrosis factor (TNF)-alpha in nasal lavage fluid were measured using enzyme-linked immunosorbent assays (ELISAs). Lung tissue and nasal mucosa sections were stained with Mayer's hematoxylin and eosin for assessment of cell infiltration and mucus production, Immunohistochemistry, Real-time PCR and Western Blot analyses for HO-1 protein expression.@*RESULT@#Orally administered Xuebijing significantly inhibited the number of OVA-induced inflammatory cells and IgE production, along with reduced T-helper (Th) 2 cytokine levels, such as IL-4, IL-5 and IL-13, improved the level of IFN-gamma, in nasal lavage fluid. In addition, Xuebijing induced a marked decrease in OVA-induced inflammatory cell infiltration and mucus production in nasal and lung tissues. These effects were correlated with HO-1 mRNA and protein induction.@*CONCLUSION@#Our results indicate that Xuebijing protects against OVA-induced airway inflammation, at least in part, via HO-1 upregulation.
Subject(s)
Animals , Mice , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Eosinophilia , Metabolism , Heme Oxygenase-1 , Metabolism , Immunoglobulin E , Allergy and Immunology , Inflammation , Drug Therapy , Metabolism , Interleukin-13 , Metabolism , Interleukin-4 , Metabolism , Interleukin-5 , Metabolism , Membrane Proteins , Metabolism , Nasal Mucosa , Metabolism , Rhinitis, Allergic , Rhinitis, Allergic, Perennial , Drug Therapy , Metabolism , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To determine the origin of 1 prenatally detected small supernumerary marker chromosome (sSMC) using SNP-chip technology, and to deduce the underlying mechanism.</p><p><b>METHODS</b>The fetal sample was subjected to karyotype analysis. The identified sSMC was subjected to genom wide scan using a SNP microarray chip. The results were validated with fluorescence in situ hybridization (FISH).</p><p><b>RESULTS</b>The karyotype of the fetus was determined as 46, X, +mar, which was verified by SNP microarray chip analysis as Yp11.2-11.3 duplication, along with loss of Yq11.2 region, FISH analysis has confirmed that the sSMC has derived from the Y chromosome.</p><p><b>CONCLUSION</b>The karyotype of the fetus was determined as 46, X, idic(Y) (pter→ p11.2::11.2→ pter). Regional deletion of Yq11.2 has been associated with male azoospermia. SNP chip analysis can exclude minor deletions and duplications with a size of more than 1 Mb, which may be applied for verifying difficult cases as well as microdeletion and duplication syndromes upon prenatal diagnosis.</p>
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Adult , Female , Humans , Male , Pregnancy , Chromosome Disorders , Diagnosis , Embryology , Genetics , Genetic Markers , Genetics , Karyotyping , Oligonucleotide Array Sequence Analysis , Polymorphism, Single Nucleotide , Prenatal DiagnosisABSTRACT
Objective To detect the changes on the expression of putative drug effiux genes caused by isoniazid-inducement in single resistance to the isoniazid Mycobacterium tuberculosis (M.tuberculosis) clinical isolates,for exploring the putative effiux genes which causing M.tuberculosis isoniazid resistance as well as the mechanism related to high expression of the putative effiux genes.Methods We selected 35 M.tuberculosis clinical isolates which were only resistant to isoniazid as well as 10 sensitive M.tuberculosis clinical isolates and using H37Rv as control.Each strain was cultured in 7H9 liquid medium without isoniazid and with subinhibitory isoniazid concentration (1/4 MIC) induction.After RNA extraction and reverse transcription,real-time PCR was conducted to assess the expression changes of 27 putative drug effiux pump genes with formula 2-△△CT to calculate the expression of each putative drug efflux pump genes.Results Of the 27 putative genes,13 of them were expressed at high level.High expression of Rv1258c gene had the maximum number of 6strains,followed by high expression of Rv0849 and Rv2265 which both had 5 strains.Fourteen strains (40.00％) out of the 35 strains had high expression pump genes.Six strains (17.14％) had only one highly expressed putative efflux pump gene.Eight strains (22.86％) had two or more highly expressed putative effiux pump genes,including two,four,five,seven genes that highly expressed in 4,2,1,1strains respectively.For the 27 putative genes,ten sensitive strains and H37Rv did not show highly expressed genes.Conclusion Rv1258c,Rv2265,Rv0849,etc.genes might be the putative effiux pumps genes of M.tuberculosis resistant to isoniazid.Isoniazid might serve as the inducer of M.tuberculosis part putative effiux pump genes,inducing activation and causing high expression of these putative effiux pump genes.
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<p><b>OBJECTIVE</b>Development and application of a real time fluorescent quantitative PCR (FQ-PCR) assay for detecting WU polyomavirus in children with low respiratory tract infections.</p><p><b>METHODS</b>The VP2 gene of WU polyomavirus was selected as the detection target, from which the real time primers and probes were designed. The standard curve was established by using recombinant plasmid as template. And the FQ-PCR assay for specific detection of WU polyomavirus was established. The specificity, sensitivity and reproducibility of the method were evaluated. Furthermore, the clinical specimens from children with respiratory tract infections collected in Wenling First People's Hospital were quantitatively detected using this method.</p><p><b>RESULTS</b>In this study, the FQ-PCR method was established to detect a specific fragment in VP2gene of WU polyomavirus. The standard curve coefficient R2 was 0.998. And this method can detect as low as 50 copies recombinant plasmid. The clinical specimens of sputum and throat swab from children with respiratory tract infections were quantitatively detected using this method. 7 sputum specimens were detected as WU polyomavirus positive in 700 sputum specimens, the positive ratio was 1.00%. No positive specimens were detected in 146 specimens of throat swabs and 846 blood samples from same patient population.</p><p><b>CONCLUSION</b>The results indicated that the FQ-PCR assay method established in this study was specific, rapid and sensitive for detecting WU polyomavirus in children with lower respiratory tract infections. The sputum specimen is more suitable to be used for gene detection of WU polyomavirus than throat swab or blood.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Polyomavirus , Real-Time Polymerase Chain Reaction , Methods , Respiratory Tract Infections , Virology , Sputum , VirologyABSTRACT
To develop polyclonal antibodies against predicted B cell epitopes in HIV-1 accessory protein Vpr, the prepared consensus Vpr amino acid sequence was used to predict potential B cell epitopes by online softwares (ABCpred and Bcepred), the synthesized polypeptides of B-cell epitopes were subsequently conjugated with keyhole limpet hemocyanin (KLH) and then used to immunize rabbits. The antibody titers were determined by ELISA, and antibody specifity was analyzed by Western-Blotting and immunoprecipitation, respectively. Amino acid residues 3-19 (N) and 82-95 (C) of Vpr were predicted as the potential B cell epitopes. After inoculation of the conjugation of synthesized peptide to KLH, the antibody titers in rabbit sera against N and C peptides reached more than 1:100000 by ELISA. Western-Blotting analysis showed that the polyclonal antibodies reacted with both wild Vpr and fusion protein of GFP with Vpr, no matter Vpr was derived from HIV-1 B subtype or CRF07_BC recombinant form; Immunoprecipitation analysis showed similar reactions to Western-Blotting results. Two B cell epitopes of Vpr were successfully predicted by Bio-informatics methods and polyclonal antibodies against those peptides could be successfully prepared.