1.Rapid detection of clinical common bacteria using DNA microarray
Guanhua DENG ; Xuan ZHENG ; Yimin HU ; Songmei LIU ; Haibo MA ; Yan XIE ; Xin ZHOU
Chinese Journal of Laboratory Medicine 2011;34(11):1012-1016
Objective To detect eight kinds of clinical common pathogenic bacteria by DNA microarray.Methods Eight kinds of common pathogenic bacteria,including Staphylococcus aureus,Pseudomonas aeruginosa,Klebsiella pneumoniae,Escherichia coli,Proteus mirabilis,Enterobacter aerogenes,Pseudomonas fluorescens,Shigella sonnei were collected.Universal primers were designed to amplify 16S rRNA gene fragment from the genomic DNA of the eight bacteria,and probes were designed in the highly variable regions.DNA microarray detection system was established and used for detection of colleted bacteria.A total of 50 samples were collected from the Zhongnan Hospital of Wuhan University,including 6 blood samples,32 sputum samples,9 feces samples and 3 bronchoscope lavage samples.DNA were extracted and detected by the established DNA microarray system.Results The desired fragments were well amplified by the self-designed universal primers.The selected probes had good detection results according to repeated detection.Of the 50 samples detected,pathgenic bacteria were accurately detected in 47 samples.Other three samples were not detected as those bacteria were not included in the chip.By optimizing the detection process,the results could be reported within 8 hours.Observation of probe signal attenuation indicated that even attenuated after 60 days,but the attenuation did not affect the results.Conclusion A microarray system was established for detection of clinical common bacteria accurately and quickly,which provided foundation for its clinical application.
2.Expression and significance of fgl2 prothrombinase in cardiac microvascular endothelial cells of rats with type 2 diabetes.
Yanping, DING ; Kun, LIU ; Yan, WANG ; Guanhua, SU ; Heping, DENG ; Qiutang, ZENG ; Yuhua, LIAO ; Zhaohui, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):575-81
Microthrombosis may be involved in the pathogenesis of cardiac microangiopathy due to diabetes. Recent studies have shown that fibrinogen-like protein 2 (fgl2) plays a pivotal role in microthrombosis in viral hepatitis, acute vascular xenograft rejection and cytokine-induced fetal loss syndrome. The current study was designed to examine the expression of fgl2 in microvascular endothelial cells and investigate the effects of microthrombi due to fgl2 on cardiac function and structure in rats with type 2 diabetes. Following induction of type 2 diabetes, 24 rats were observed dynamically. Fgl2 expression and related cardiac microthrombosis were examined. Local or circulating TNF-α was measured. Coronary flow (CF) per min was calculated as an index of cardiac microcirculation. Cardiac function and morphology were evaluated. It was found that Fgl2 was highly expressed in cardiac microvascular endothelial cells of rats with type 2 diabetes, which was promoted by local or circulating TNF-α. The Fgl2 expression was associated with cardiac hyaline microthrombosis. In parallel with the fgl2 expression, CF per min, cardiac diastolic or systolic function and cardiac morphology were aggravated to some extent. It was concluded that in rats with type 2 diabetes, microthrombosis due to fgl2 contributes to the impairment of cardiac diastolic or systolic function and morphological changes.
3.Prediction of microvascular invasion based on enhanced mode magnetic resonance imaging for patients with hepatocellular carcinoma
Wenjie SUN ; Zhiling GAO ; Guanhua YANG ; Yujia GAO ; Jing JIA ; Haijing QIU ; Lin DENG ; Yong CHEN
Chinese Journal of Hepatobiliary Surgery 2021;27(3):175-180
Objective:To study preoperative MRI imaging and its enhanced mode on tumor features in predicting microvascular invasion (MVI) in patients with hepatocellular carcinoma (HCC).Methods:The clinical data of patients with a solitary HCC who underwent MRI examination followed by surgical resection at the General Hospital of Ningxia Medical University from January 2017 to June 2019 were studied. The patients were divided into the MVI (+ ) and MVI (-) groups according to the findings on postoperative pathological diagnosis. The relationship between the rates of MVI and MRI tumor features including diffusion weighted imaging (DWI) signal, enhancement mode, enhancement type and other imaging characteristics were analysed.Results:Of 84 patients with HCC enrolled into this study, there were 65 males and 19 females. Their age (Mean±SD) was (54.94±11.51) years. MVI (+ ) was found in 46 patients and MVI (-) in 38 patients. The maximum tumor diameters (Mean±SD) of the two groups were (7.08±3.45) cm and (4.28±2.47) cm ( P<0.01). Single-factor analysis and comparison of imaging characteristics of the two groups of patients showed tumor DWI signal, tumor encapsulation, enhancement mode, tumor edge smoothness, abnormal enhancement around tumors, and intratumoral arteries were significantly different ( P<0.05); There were no significant differences in T 1WI signals, T 2WI signals, tumor periphery, and enhancement types between groups. After inputting MVI(+ ) as a risk factor into the logistic regression model, tumor maximum diameters >6.33 cm, type 3/4 enhancement mode, and unsmoothness of tumor edge were independent risk factors (all P<0.05). Through combined diagnosis using ROC curve analysis with a cut-off value of 0.53, the area under the curve was 0.881, the sensitivity 0.870, specificity 0.789, and the Youden index 0.659. Conclusion:The multivariate logistic regression model and combined diagnosis using ROC curve analysis improved the diagnostic efficacy of MVI in its prediction of HCC on imaging studies. The risk predictors were easy to use and to promote in clinical practice.
4.The establishment of a gene microarray for detecting the common pathogenic fungi
Guanhua DENG ; Xuan ZHENG ; Xin ZHOU ; Yimin HU ; Yan XIE ; Songmei LIU
Chinese Journal of Laboratory Medicine 2011;34(12):1114-1118
Objective To develop a gene microarray system for detection of clinical common pathogenic fungi.MethodsThere were 8 clinical common fungi chosen as the subjects including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilokis,Candida pseudotropicalis,Aspergillus terreus,Aspergillus flavus,Aspergillus oryzae,Aspergillus fumigatus.Universal primers,probes and specific probes for the PCR amplification and microarray preparation were designed in ITS region of the fungi genomic DNA.The PCR products amplified from those fungi's genome DNA were denatured and hybridized with the probes in gene microarray.The rapid detection of fungi was based on the investigation on the fluorescent signal intensity in the chip.The detection results of gene microarray system were verified by true positive and negative clinical samples.Results There were totally 25 positive samples identified by clinical routine microbiological methods.The 10 samples identified as bacteria positive were determined as negative without fluorescent signal by the fungi gene microarray,while the 12 samples identified as fungi positive were determined as positive with certain fungus by the fungi gene microarray.And 3 artificial Candida krusei samples were detected as fungi positive,while they were failure to be identified as certain fungus.There was no fluorescent signal in positions of the 8 fungi specific probes,but there was fluorescent signal in the position of fungi universal probe.It indicated that there were fungi in the samples but it couldn't identify the species of the fungi,because the Candida krusei wasn't included in the detection fungi list of the fungi gene microarray.ConclusionsThe fungi gene microarray established by the study could detect the common fungi in clinic rapidly and accurately.This study lays technology foundation for clinical application of gene chip.
5.Expression and Significance of fgl2 Prothrombinase in Cardiac Microvascular Endothelial Cells of Rats with Type 2 Diabetes
DING YANPING ; LIU KUN ; WANG YAN ; SU GUANHUA ; DENG HEPING ; ZENG QIUTANG ; LIAO YUHUA ; WANG ZHAOHUI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):575-581
Microthrombosis may be involved in the pathogenesis of cardiac microangiopathy due to diabetes. Recent studies have shown that fibrinogen-like protein 2 (fgl2) plays a pivotal role in microthrombosis in viral hepatitis, acute vascular xenograft rejection and cytokine-induced fetal loss syndrome. The current study was designed to examine the expression of fgl2 in microvascular endothelial cells and investigate the effects of microthrombi due to fgl2 on cardiac function and structure in rats with type 2 diabetes. Following induction of type 2 diabetes, 24 rats were observed dynamically. Fgl2 expression and related cardiac microthrombosis were examined. Local or circulating TNF-a was measured. Coronary flow (CF) per min was calculated as an index of cardiac microcirculation. Cardiac function and morphology were evaluated. It was found that Fgl2 was highly expressed in cardiac microvascular endothelial cells of rats with type 2 diabetes, which was promoted by local or circulating TNF-α. The Fgl2 expression was associated with cardiac hyaline microthrombosis. In parallel with the fgl2 expression, CF per min, cardiac diastolic or systolic function and cardiac morphology were aggravated to some extent. It was concluded that in rats with type 2 diabetes, microthrombosis due to fgl2 contributes to the impairment of cardiac diastolic or systolic function and morphological changes.
6.Multi-slice CT imaging features as risk factors in determining spontaneous rupture of primary liver cancer
Kun LIN ; Zhiling GAO ; Guanhua YANG ; Lin DENG ; Tao REN ; Yong CHEN
Chinese Journal of Hepatobiliary Surgery 2021;27(9):658-662
Objective:To analyze the imaging features of spontaneous rupture of primary liver cancer (PLC) and to study the high-risk factors associated with tumor rupture.Methods:From September 2016 to August 2020, 81 patients who developed spontaneous rupture of PLC at the General Hospital of Ningxia Medical University were included into this study. A control group of 81 patients with tumors located on the periphery of the liver but without rupture treated in the same period were selected by matching the two groups with age, sex and BCLC staging. The clinical data and CT imaging characteristics including tumor location, extent, size, and morphology of the two groups of patients were compared retrospectively between groups.Multivariate logistics regression was used to analyze.Results:A total of 81 patients were included in the case group, including 72 males and 9 females, aged (53.69±10.34) years. The control group included 81 patients, 64 males and 17 females, aged (54.78±9.04) years. The main risk factors for spontaneous rupture of PLC included in this study were cirrhosis, tumor close to diaphragm, biolobar distribution, portal vein obstruction, tumor diameter >10 cm, invasion of liver capsule (arc-to-chord ratio>1) and tumor protrusion ≥25% ( P<0.05). Logistic regression analysis showed that cirrhosis ( OR=2.796, 95% CI: 1.721-10.834), portal vein obstruction ( OR=3.586, 95% CI: 1.272-10.107) and tumor protrusion (≥25%) ( OR=2.831, 95% CI: 1.668-22.210) were independent predictive factors of spontaneous rupture of PLC. Conclusion:Tumor protrusion≥25%, cirrhosis and portal vein obstruction were closely related to spontaneous rupture of PLC. They were independent risk factors in predicting rupture of primary liver cancer.
7.Prediction of the degree of differentiation of hepatocellular carcinoma before surgery based on clinical data and MRI image features
Lin DENG ; Zhiling GAO ; Wenjie SUN ; Tao REN ; Guanhua YANG ; Yujia GAO ; Haijing QIU ; Yong CHEN
Chinese Journal of Hepatobiliary Surgery 2021;27(7):499-504
Objective:To explore the value of clinical data and MRI image features in predicting and analyzing the degree of differentiation of hepatocellular carcinoma (HCC).Methods:The clinical and imaging data of 180 patients with surgical outcomes of HCC from March 2015 to June 2019 in the General Hospital of Ningxia Medical University were retrospectively analyzed. Alpha-fetoprotein (AFP)、aspartate aminotransferase (AST)、D-dimer、clinical stage、tumor length、apparent diffusion coefficient(ADC)、enhancement types and so on the clinical and imaging data of the poorly differentiated and non-differentiated HCC were compared and analyzed. Multivariate logistic regression was used to predict independent risk factors for poorly differentiated HCC.Results:Of the 180 HCC patients, 121 were moderately and highly differentiated, and 59 were poorly differentiated. Univariate analysis showed that the patient’s age, gender, AFP, AST, D-dimer level, clinical stage, Child-Pugh score, tumor length, whether the capsule was complete, tumor apparent diffusion coefficient, the maximum level ADC value, enhancement type with HCC differentiation degree were correlated(all P<0.05). Multivariate logistic regression analysis showed that the patients' gender ( OR=4.524, P<0.05), clinical stage ( OR=5.598, P<0.05), D-dimer ( OR=8.576, P<0.05), HCC diameter ( OR=0.498, P<0.05), enhancement types ( OR=2.988, P<0.05), tumour ADC value ( OR=0.059, P<0.05) were independent of poorly differentiated HCC risk factor. Conclusion:MRI image features can be used as an effective indicator to predict the degree of HCC differentiation before surgery. It is more valuable to accurately predict the degree of HCC combined with D-dimer and AFP value.
8.Chemical constituents of rhizomes of Zingiber officinale.
Lei BAO ; Anjun DENG ; Zhihong LI ; Guanhua DU ; Hailin QIN
China Journal of Chinese Materia Medica 2010;35(5):598-601
OBJECTIVETo investigate the chemical constituents from the rhizomes of Zingiber officinale.
METHODIsolation and purification of the chemical constituents were carried out on the column chromatography of silica gel and Sephadex LH-20. The structures were elucidated on basis of physicochemical properties and spectral data.
RESULTTen compounds were isolated and identified as beta-sitosterol palmitate (1), isovanillin (2), glycol monopalmitate (3), hexacosanoic acid 2,3-dihydroxypropyl ester (4), maleimide-5-oxime (5), p-hydroxybenzaldehyde (6), adenine (7), 6-gingerol (8), 6-shogaol (9), and 1-(omega-ferulyloxyceratyl) glycerols (10a-10f).
CONCLUSIONCompounds 1-7 and 10a-10e are obtained from Z. officinale for the first time, and compound 10f is a new compound.
Ginger ; chemistry ; Rhizome ; chemistry
9.Exclusive control substance of radix Stephaniae tetrandrae.
Ying LIU ; Anjun DENG ; Xifeng LI ; Zhihong LI ; Jinlan ZHANG ; Guanhua DU ; Hailin QIN
China Journal of Chinese Materia Medica 2009;34(15):1943-1948
OBJECTIVETo develop the system for the exclusive control substance of plant drug (CSPD) in traditional Chinese herbal medicines (TCHM), this paper investigated the (CSPD) of Radix Stephaniae Tetrandrae as well as its proton nuclear magnetic resonance (1H-NMR) and high performance liquid chromatography (HPLC) analytical methods for the purpose of original identification and quality control of the crude drug.
METHODThe CSPDs and their 1H-NMR and HPLC profiles of Radix Stephaniae Tetrandrae were obtained by standardized procedure. Chemical components were isolated from the CSPD by silica gel column chromatography. The assignments of the characteristic signals in 1H-NMR and HPLC profiles were achieved on the basis of elucidation of the isolates structures.
RESULTFor nine samples from the different sources in this paper, the 1H-NMR and HPLC profiles from eight sources had wonderful reproducibility and characteristics, and the other gave differences compared with the eight samples in the signal strength of the main components. Furthermore, seven compounds were isolated from CSPD and their chemical structures were authenticated by spectral analysis as tetrandrine, fangchinoline, tetrandrine-2'-N-beta-oxide, tetrandrine-2'-N-alpha-oxide, dicentrine, dicentrinone, and adenine, respectively. The 1H-NMR and HPLC profiles of the CSPD of Radix Stephaniae Tetrandrae showed mainly the characteristic signals of the bisbenzylisoquinoline alkaloids isolated in this work.
CONCLUSIONThe 1H-NMR and HPLC profiles of the CSPD of Radix Stephaniae Tetrandrae exhibit the structures and total composition of the main active constituents in it, and can be used for its original identification and quality evaluation.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Quality Control ; Stephania tetrandra ; chemistry
10.Analysis of radiation doses from head and chest CT scanning for children in Ningxia
Haijing QIU ; Zhiling GAO ; Jing JIA ; Yujia GAO ; Tao REN ; Wenjie SUN ; Guanhua YANG ; Lin DENG ; Yong CHEN
Chinese Journal of Radiological Medicine and Protection 2020;40(11):851-856
Objective:To evaluate the radiation doses from head and chest CT examinations of children in Ningxia, and provide basic data for the optimization of CT radiation doses to children of different ages.Methods:By using stratified cluster sampling method , the dose parameters on head and chest CT scanning, CTDI vol and DLP for the children under 15 years old were sampled within 1-2 weeks from hospitals at different levels in different regions of Ningxia for the calculation of effective doses. The 75th percentile (P75) of CTDI vol and DLP was compared with those recommended by other countries. All children included four age groups: up to 1 year old, 1 to 5 years, 6 to 10 years, 11 to 15 years. Results:There were 39 hospitals and 47 CT scanners in this survey, and 1 134 head scanning and 636 chest scanning were investigating. The 75th percentile (P75) of CTDI vol and DLP for head scanning were 44.2 mGy and 456.2 mGy·cm for those up to 1 year old; 57.2 mGy and 659.6 mGy·cm for 1 to 5 years old, 61.1 mGy and 668.7 mGy·cm for 6-10 years old, and 63.6 mGy and 849.3 mGy·cm for 11-15 years old, respectively. The 75th percentile (P75) of CTDI vol and DLP for chest scanning were 5.0 mGy and 89.2 mGy·cm for those up to 1 year old, 5.9 mGy and 124.8 mGy·cm for 1 to 5 years old, 6.0 mGy and 167.9 mGy·cm for 6 to 10 years, and 7.1 mGy and 235.0 mGy·cm for 11 to 15 years old, respectively. Conclusions:The chest CT radiation dose to children in Ningxia is close to the reported values, but the head CT radiation dose is relatively high in all age groups, especially in infants. The optimization and regulation of head CT radiation doses to children in Ningxia should be strengthened. It is high time to increase dose awareness for pediatricians and radiologists and raise awareness of radiation-related risks.