1.QUANTITATIVE DETECTION OF BACILLUS ANTHRACIS BY SYBR GREEN I REAL-TIME POLYMERASE CHAIN REACTION
Suhong CHEN ; Minli ZHANG ; Hang MOU
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Anthrax is a fatal infectious disease of human and livestock and is caused by Bacillus anthracis. To establish a simple and specific assay for the clinical diagnosis of B. Anthracis infection, two oligonucleotide primers specific for the cap region of plasmid pXO2 and two specific for the pag region of plasmid pXO1 were designed and synthesised for polymerase chain reaction (PCR). The fluorescence dye SYBR Green I incorporated into the double strands was used to quantitate, and the specificity of PCR product was confirmed by the melting curve. The results showed that 0.8?mol/L primers and 3.0mmol/L Mg 2+ were optimal for this quantitative PCR assay. The sensitivity of this assay was 10 3 copies per millitier B. anthracis could be specifically distinguished from other B. cereus group of bacteria. with this assay. SYBR Green I real-time PCR appeared to be a rapid, accurate and specific methad for quantitative analysis of B. anthracis.
2.Comparative study of arterial vulnerable plaque molecular imaging with novel molecular probes 99Tcm-Duramycin and 99Tcm-RGD in rabbit models
Ying ZHANG ; Tiantian MOU ; Qian WANG ; Jie HAN ; Hang SU ; Shanshan GU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(5):289-293
Objective To investigate the potential of 99Tcm-Durarnycin and 99Tcm-RGD in detecting vulnerable plaque in rabbit models.Methods Fifteen healthy New Zealand male rabbits were randomly divided into group A (control group,n =5),group B (stable plaque group,n =5) and group C (vulnerable plaque group,n =5).Animals were injected with 99Tcm-Duramycin and 99Tcm-RGD at the end of 4,8 and 12 weeks.SPECT/CT scanning was performed at 0.5 h post injection.One rabbit was sacrificed at the end of 4 weeks and one at the end of 8 weeks after imaging.The others were sacrificed at the end of 12 weeks after imaging studies.All aortas were collected.Intravascular ultrasound (IVUS) was performed at the end of 8,12 weeks before SPECT/CT scanning.The data was analyzed with paired t test.Results In group A,the aortas had little uptake of the two probes.In group B,the aortas showed obvious radioactive uptake of 99Tcm-Duramycin and 99Tcm-RGD at the end of 8 weeks and 12 weeks,while 99Tcm-Durarnycin gave better display than 99Tcm-RGD.In group C,99Tcm-Duramycin uptake was higher than 99Tcm-RGD uptake in the aorta.The T/NT ratios of 99Tcm-Duramycin and 99Tcm-RGD in group C were 2.14±0.34 and 1.46±0.34 (t=4.072,P<0.05) at the end of 4 weeks,2.93±0.41 and 1.66±0.22 (t=5.578,P<0.05) at the end of 8 weeks,3.25±0.29 and 1.81±0.28 (t=19.692,P<0.05) at the end of 12 weeks.In isolated specimen of group C,the yellow lipid plaque of the intima bulged on the lumen at the end of 12 weeks.IVUS indicated that,at the end of 8 weeks and 12 weeks,the endometrial thickness of group C was (450±104) mm and (767±52) mm (t=44.024,P < 0.05) respectively,and the rates of luminal stenosis were (29.30± 2.81) % and (37.98 ±6.41)% (t =9.226,P<0.05).Conclusions Both 99Tcm-Duramycin and 99Tcm-RGD may be used to detect vulnerable plaque at early time.99Tcm-Duramycin may detect vulnerable atherosclerotic plaque earlier than 99Tcm-RGD and provide better diagnostic image.
3.Immunofecal occult blood test predicts the prognosis of patients with chronic kidney disease
Hui ZHOU ; Shan MOU ; Haiyun YANG ; Qin WANG ; Beili SHI ; Leyi GU ; Ying HANG ; Zhaohui NI
Chinese Journal of Nephrology 2012;28(6):429-434
Objective To evaluate the value of immunofecal occult blood test (IFOBT) as a prognostic indicator in CKD patients with colorectal impairment.Methods A total of 176CKD patients and 180 healthy adults as control were enrolled.Serum biochemistry was measured at baseline and gastrointestinal bleeding was determined by IFOBT.All the CKD patients were followed up for 4.5 years.Renal replacement therapy or death was defined as end-point event.The Logistic regression analysis was used for risk factors.Kaplan-Meier analysis and COX regression model were used for survival analysis.Results The positive rate of IFOBT in CKD patients was significantly higher than healthy control (17% vs 5.3%,χ2=13.236,P<0.01).When comparing with IFOBT negitive patients,IFOBT positive patients were older [(62.030±15.544) years old vs (48.660±19.018)years old,P<0.01],had higher ESR [(71.800±31.657) mu/h vs (57.210±32.712) mm/h,P<0.05],C-reactive protein [6.230 (3.000~14.148) mg/L vs 3.000 (3.000~6.833)mg/L,P<0.05],serum creatinine [419.100 (103.200~546.625) μmol/L vs 175.100 (68.150~462.950) μmol/L,P<0.05],and had lower hemoglobin level [(97.970±20.590) g/L vs (107.170±27.988) g/L,P<0.05] and eGFR [11.400 (8.671~53.544) ml·min1·(1.73 m2)1 vs 35.274(10.961~82.145) ml·min-1·(1.73 m2)-1,P<0.01].There was a negative correlation between IFOBT value and eGFR in CKD patients (r=-0.20,P<0.01).Positive correlations of IFOBT value with age (r=0.175,P<0.05) and serum creatinine (r=0.171,P<0.05) were found.Logistic regression and COX regression analysis showed that IFOBT value,eGFR and ESR were important factors that influenced the prognosis of CKD patients.Kaplan-Meier analysis revealed that IFOBT value >100μg/L predicted progression of renal function.Conclusions The prevalence of gastrointestinal bleeding disorder is high in patients with CKD.Value of IFOBT independently predicts decline in renal function of CKD patients.
4.Targeting vulnerable plaque of experimental atherosclerosis models with 99Tcm-Duramycin
Shanshan GU ; Hongzhi MI ; Qian WANG ; Hang SU ; Ying FU ; Tiantian MOU ; Jie HAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(1):53-58
Objective To investigate the value of 99Tcm-Duramycin SPCET/CT in the detection of vulnerable plaque (VP) in atherosclerotic (AS) rabbit models.Methods Sixteen New Zealand male rabbits were randomly divided into three groups:group A (sham-operated group,n =4),group B (stable plaque group,n =4) and group C (VP group,n =8).Group A was fed on normal diet,and the other 2groups were fed on cholesterol diet for 12 weeks.Femoral artery dissection sham-operation was done in group A and group B,while balloon-induced abdominal aorta wall injury was produced in group C after 2 weeks'feeding.Animals were injected with 99Tcm-Duramycin (74 MBq/kg) and then SPECT/CT imaging was performed at the end of 4,8,and 12 weeks,respectively.Abdominal aortas were explanted for ex vivo imaging and histological characterization of plaque.The apoptosis index (AI) was calculated.One-way analysis of variance was used to analyze data.Results There was no radioactive uptake by the abdominal aorta in each group at the end of 4 weeks and no uptake in group A and group B at the end of 8 weeks.There was slightly uptake radioactive uptake by the abdominal aorta in group B at the end of 12 weeks and in group C at the end of 8 weeks.There was intense uptake at the lesions of AS rabbits in group C at the end of 12 weeks,and the T/NT value significantly higher than that of the other two groups (3.40±0.22 vs 2.12±0.65,2.68±0.18,F=198.775,P<0.05).The result was confirmed in the ex vivo imaging of the explanted aorta.The AI of group C was significantly higher than that of group A and B ((25.4±6.32) % vs (0±0.02)%,(5.3± 1.97)%,F=70.260,P<0.05).Conclusions 99Tcm-Duramycin scimigraphy could identify the apoptosis of VP in the rabbit AS models.It is a promising non-invasive method to diagnose AS plaques.
6.Feasibility of 99Tcm-3P4-RGD2 SPECT/CT imaging in evaluation of arterial plaque stability after atorvastatin intervention in rabbits
Jie HAN ; Ying ZHANG ; Qian WANG ; Hongzhi MI ; Hang SU ; Tiantian MOU ; Xiaofen XIE ; Quan LI ; Yehong ZHANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(7):410-414
Objective To investigate the feasibility of a novel molecular probe 99Tcm-3P4-RGD2 in evaluating arterial plaque stability after atorvastatin intervention in rabbits with SPECT/CT. Methods Eighteen male New Zealand rabbits were randomly divided into group A (stable plaque), group B (vulnerable plaque), and group C (vulnerable plaque with statin intervention). All rabbits were fed with high-fat food for 12 weeks. After high-fat feeding for two weeks, sham surgery was performed on group A. In the meantime, abdominal aorta injury was performed on group B and group C. After that, rabbits of group C were given oral atorvastatin (2.5 mg·kg-1·d-1). 99Tcm-3P4-RGD2 SPECT/CT imaging was performed on each group at the end of 4, 8 and 12 weeks. T/NT ratios were calculated. Animals were sacrificed at the end of 12 week after imaging studies. The abdominal aortas were collected, imaged with SPECT/CT, and evaluated by pathological HE staining and immunohistochemical analysis. MVD was calculated. Differences among 3 groups were analyzed using one-way analysis of variance. Results There was no significant radioactive uptake in the abdominal aortas of three groups on the 4th week′s imaging. The radioactive uptake in abdominal aortas increased slightly on the 8th week, with the highest radioactive uptake in group B. The radioactivity in abdominal aortas of the 3 groups maintained increasing on the 12th week, with T/NT ratios of 1.579±0.217, 1.873±0.226 and 1.524±0.237, respectively (F=8.984, P<0.05). In ex vivo abdominal aorta images, especially images of group B, radioactivity in lesion sites was higher than that in normal tissue. Accordingly, results of HE staining showed that artery plaques of group A and group C were grade Ⅱ and group B was grade Ⅳ. The MVD of group A, B and C was 8.17±1.17, 15.86±1.07 and 7.17±1.60, respectively (F=9036, P<0.05). Conclusion 99Tcm-3P4-RGD2 SPECT/CT imaging has a high sensitivity in the evaluation of arterial plaque stability after statin intervention in rabbits.
7.Analysis of clinicopathology and plasmapheresis efficacy in patients with anti-glomerular basement membrane disease
Lijun MOU ; Limeng CHEN ; Laimeng ZUO ; Yubing WEN ; Hang LI ; Yon QIN ; Mingxi LI ; Jianling TAO ; Wenling YE ; Hong XU ; Wei YE ; Yang SUN ; Xuemei LI ; Xuewang LI
Chinese Journal of Nephrology 2011;27(4):230-235
objective To analyze the clinicopathological features and prognosis of antiglomerular basement membrane(GBM)disease,and evaluate the efficacy and safety of double filtration plasmapheresis(DFPP). Methods A total of 35 hospitalized patients diagnosed as anti-GBM disease in our department were enrolled in the study.All the patients were divided into 3 groups according to the manifestations at admission.Group Ⅰ∶24 patients with severe pulmonary hemorrhage or rapidly progressive glomerulonephritis(RPGN)received pulse methylprednisolone with or without DFPP,and then followed by prednisone and CTX.Group Ⅱ∶5 patients without severe pulmonary hemorrhage and RPGN received prednisone and CTX.Group Ⅲ∶5 ESRD patients and 1 normal renal function patient did not receive immunosuppression therapy.Anti-GBM antibody titer of pre-and post-DFPP in 4 patients was measured consecutively,and removal rate was calculated.Results The mean age of all the patients was(41.1±16.6)years.Sixteen patients(45.7%)presented Goodpasture's syndrome.Eighteen patients(51.4%)had anti-GBM glomerulonephritis alone,whereas one suffered solely from pulmonary hemorrhage.20%patients had positive P-ANCA serology.54.2%crescentic glomerulonephritis and 7 with other glomerulonephritis were revealed by kidney biopsy in 24 patients.Patients in Group Ⅰ showed more severe manifestation at admission:higher Scr level,higher titer of anit-GBM antibody,greater percentage of crescents.Within the follow-up period,7 patients died and kidneys of 50%patients survived.No patient died in Group Ⅱ and Ⅲ.The elder age,anemia,higher Scr(>300 μmol/L),oliguria or anuria,emergency hemodialysis at admission,and more glomerular sclerosis were predictors of poor prognosis.The anti-GBM antibody was negative after 4 to 6 sessions of DFPP.and the mean removal rate was 55%.During total 94 DFPP sessions,there was no unacceptable morbidity. Conclusions Different therapy strategy is necessary for anti-GBM disease with different clinical manifestations.DFPP is an effective and safe clearance way of anti-GBM antibody.
8.The relationship between urinary albumin excretion and serum uric acid in general population
Xiaohong FAN ; Jianfang CAI ; Bixia GAO ; Lijun MOU ; Jinhong LI ; Xuejiao LIU ; Junxue WU ; Qingyan MENG ; Haiyun WANG ; Lili LIU ; Hang LI ; Xuemei LI ; Xuewang LI
Chinese Journal of Internal Medicine 2011;50(7):550-554
Objective To investigate the relationship between the urinary albumin excretion (UAE) and serum uric acid in general population. Methods The study participants were derived from the epidemiological study on the association of metabolic syndrome and chronic kidney disease (CKD) in Pinggu district, Beijing. A total of 992 participants (463 men and 529 women) aged from 30 to 75 years were enrolled in this study. For each participant, UAE, serum uric acid, serum creatinine, and serum lipids were detected and other potential risk factors for CKD were surveyed. Results ( 1 ) The frequencies of microalbuminuria, macroalbuminuria and hyperuricemia were 12.9% , 1.8% and 4.3% respectively. The persons with hyperuricemia had significantly higher frequency of albuminuria than those without hyperuricemia (37. 2% vs 13. 7% , P <0. 01). (2) The participants were divided according to the quartiles (25% , 50% , 75% ) of serum uric acid level, and the frequencies of albuminuria in males were 13. 2% , 13. 9% , 17. 2% and 25.4% , while those in females were 8. 4% , 6. 2% , 9. 6% and 24. 8%. ( 3 ) Multivariate logistic regression analysis showed, hyperuricemia was significantly associated with albuminuria in females (OR =2. 31, 95% CI 1. 15-4. 68; P=0.02), but not in males. If the persons with reduced renal function were excluded, similar result still could be gained. Conclusions The prevalence of albuminuria increases gradually with uric acid elevation. Serum uric acid is an independent risk factor of elevated UAE, especially in females.
9.Discriminator value of Han Chinese albumin creatinine ratio for detecting microalbuminuria
Xiaohong FAN ; Jianfang CAI ; Bixia GAO ; Lijun MOU ; Jinhong LI ; Xuejiao LIU ; Junxue WU ; Qingyan MENG ; Haiyun WANG ; Lili LIU ; Hang LI ; Xuemei LI ; Xuewang LI
Chinese Journal of Nephrology 2010;26(11):807-811
Objective To investigate the discriminator value of Han Chinese first morning urine albumin creatinine ratio (ACR) for determining the microalbuminuria. Methods A total of 1056 participants (494 males and 562 females) were selected from epidemiologic study of the metabolic syndrome and chronic kidney disease in Pinggu district, Beijing. Eight-hour overnight urinary albumin excretion (UAE) was regarded as the gold standard for defining the albuminuria,and the ROC curve analysis was used to determine the ACR discriminator value for microalbuminuria. Results (1)Microalbuminuria was found in 12.5% of participants,macroalbuminuria in 1.7%. (2)The ACR discriminator value for microalbuminuria by ROC curve analysis was 1.95 g/mol (sensitivity 97.6% and specitivity 88.6%) for men, 3.62 g/mol(sensitivity 83.8% and specitivity 89.1%) for women, 2.78 g/mol (sensitivity 88.7% and specitivity 85.9%)for overall. The upper boundary of microalbuminuria by ROC curve analysis was 22.59 g/mol (sensitivity 100.0% and specitivity 98.8%) . (3)The inter-rater agreement of the result in this study showed that sensitivity was 91.3% and specitivity was 88.2%, positive likelihood ratio was 7.56 and negative likelihood ratio was 0.10, positive predictive value was 56.9% and negative predictive value was 98.4%. Conclusions The ACR discriminator value for determining microalbuminuria is obviously higher in women than that in men, and is higher than recommendation of international guidelines. The result by ROC curve analysis has better sensitivity and specitivity.
10.Clinical analysis of anti glomerular basement membrane disease with normal renal function.
Li-jun MOU ; Li-meng CHEN ; Lai-meng ZUO ; Hai-yun WANG ; Jian-ling TAO ; Yu-bing WEN ; Wen--ling YE ; Yan QIN ; Hang LI ; Xue-mei LI ; Xue-wang LI
Acta Academiae Medicinae Sinicae 2011;33(4):432-435
OBJECTIVETo investigate the clinical and pathologic characteristics of anti-glomerular basement membrane(GBM) disease with normal renal function.
METHODSThe clinical and pathologic data of 6 patients with anti-GBM disease and normal renal function in Peking Union Medical College Hospital were reviewed retrospectively. Furthermore, 29 patients with anti-GBM disease and impaired renal function in the same period in the same hospital were enrolled as the control group. Factors that may influence the prognosis were analyzed.
RESULTSSix (17.1%) of all 35 patients maintained normal renal function for 12-133 months during follow-up. Five patients had microhematuria and proteinuria, one had pulmonary hemorrhage only, and three manifested as Goodpasture syndrome. Renal biopsies from 4 patients revealed linear deposition of IgG 2+-3+ along the glomerular capillary walls by immunofluorescence. As shown by normal light microscopy, mild mesangial proliferation and crescentic glomerulonephritis with a large amount of fibrinoid necrosis of glomerular capillary walls were observed in different patients; however, most pathological changes were mild. Five of these six patients were treated with immunosuppressive drugs and/or plasma exchange. Compared with the control group, the 6 patients with normal renal function had significantly higher hemoglobin[(77.97±20.62 vs.(99.67±19.80 g/L P=0.024], lower titers of anti-GBM antibody[(224.34 ± 145.79 vs.(80.23 ± 85.73 EU/ml P=0.027], and lower ratio of glomeruli with crescents[(0.58±0.29 vs.(0.17±0.27 ,P=0.005]. These 6 patients with normal renal function were followed up for 12-133 months, among whom 4 patients achieved complete remission and 2 had mild proteinuria and microhematuria.
CONCLUSIONAnti-GBM disease with normal renal function is not uncommon. Most patients have mild pathologic changes and good prognosis.
Adult ; Anti-Glomerular Basement Membrane Disease ; pathology ; physiopathology ; Female ; Follow-Up Studies ; Humans ; Kidney ; physiopathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies