1.Trend analysis of endemic arsenism in the monitored areas of the disease in 2010 - 2012 in Inner Mongolia
Yajuan XIA ; Kegong WU ; Kuiying FANG ; Zhiwei GUO ; Na CUI
Chinese Journal of Endemiology 2016;(1):32-34
Objectives To monitor the residents prevalence of endemic arsenism in the disease affected areas in Inner Mongolia, so as to provide feasible suggestions for control of arsenism in the future. Methods Monitoring data were obtained from the Project of Endemic Disease Prevention Granted by Central Government in 2010 - 2012, and the conditions of arsenism patients from 38 endemic arsenic villages were analyzed among different year, age and gender. Results The detection rate of arsenism of the 38 surveillance villages was 7.38%(517/7 004) in 2010, 7.10%(482/6 784) in 2011 and 6.62% (431/6 514) in 2012. The arsenism patients of mild;moderate and severe cases from 2010 to 2012, accounted for 74.47% (385/517), 74.27% (358/482), 75.17% (324/431); 16.83% (87/517), 16.60% (80/482), 15.78% (68/431) and 8.7% (45/517), 9.13% (44/482), 9.05% (39/431), respectively. For skin lesions, the detection rates of keratosis, pigmentation and depigmentation from 2010 to 2012, were 8.08%(566/7 004), 7.90%(536/6 784), 7.77%(506/6 514);3.27%(229/7 004), 3.29%(223/6 784), 2.87%(187/6 514) and 6.68% (468/7 004), 6.63% (450/6 784), 5.82% (379/6 514), respectively, showed a declining trend. It also showed a declining trend with age, and the patients were mainly 40 years old people and older, and the highest detection rate was in the 60- 70 years old group[15.54%(143/920)、14.72%(135/917)、13.36%(136/1 018)]. For gender distribution, the detection rate of the three years was higher in male than female [male 8.24%(300/3 639), 7.99%(283/3 542), 7.71%(260/3 372);female 6.45%(217/3 365), 6.14%(199/3 242), 5.44%(171/3 142),χ2=8.24, 8.77, 13.54, all P〈0.01]. Conclusion There is no big change of arsenism conditions in 2010-2012, with a slight declining trend.
2.Multiplex real-time quantitative polymerase chain reaction for simultaneous detection of Escherichia coli and Candida albicans genes in human whole blood
Jiabin FANG ; Junren KANG ; Enling MA ; Guangliang GUO ; Xizeng CUI
Chinese Journal of Clinical Nutrition 2015;23(3):170-175
Objective To establish a multiplex real-time quantitative polymerase chain reaction (MRQPCR) assay for fast and simultaneous detection of Escherichia coli (E.coli) and Candida albicans (C.albicans) genes in human whole blood,in order to facilitate differentiation of the types of microorganism and evaluation of the severity of bacterial or fungi translocation due to impaired gut barrier,hence providing help to select specific antimicrobial agents.Methods The β-D-galactosidase gene of E.coli and ITS2 gene of C.albicans were selected as the target genes for designing primers and probes.E.coli and C.albicans genomes were extracted with QIAamp(R) DNA Blood Mini Kit,and the 25 μl TaqMan MRQ-PCR amplification reaction system was established.18 simulated human whole blood samples and 10 whole blood samples from febrile surgical patients were detected for E.coli and C.albicans genes using MRQ-PCR.Results The specificity of the primers and probes were excellent.The correlation coefficients of the standard curves of E.coli and C.albicans were 0.994-0.999 and 0.994-0.998,respectively;and the efficiency of amplification were 0.894-1.022 and 0.905-1.028,respectively.In the standard samples,the lowest detection limits of E.coli and C.albicans were 13.9 copies/μl and 0.8 cfu/μl,respectively;the sensitivity was 100% and 99.69%,the specificity was 100% and 94.73%,respectively;the average recovery rates were (101.89 ± 5.69)% and (103.74 ± 4.64)% respectively;the intra-batch coefficients of variance (CV) in detecting the genes were (13.14 ± 10.27)% and (19.18 ± 8.54)%,respectively,and the inter-batch CV were (14.35 ± 9.34)% and (18.31 ± 10.25) %,respectively.In human whole blood,the lowest detection limits of E.coli and C.albicans were 12 455.2 copies/ml and 800.3 cfu/ml,respectively;the average recovery rates were (111.60 ± 11.06) % and (99.96 ± 6.16) %,respectively;the intra-batch CV in detecting the genes were (11.02 ± 5.65) % and (8.14 ± 7.29)%,respectively,and the average inter-batch CV were (12.88 ± 7.59)% and (18.62 ± 9.14)%.Conclusions MRQ-PCR is a rapid,sensitive,specific,accurate,and reproducible method for simultaneous detection of E.coli and C.albicans genes in human whole blood,with sample-,cost-,and time-saving advantages.It is a promising technique for rapid differentiation between fungi and bacteria,which could help targeted administration and evaluation of antimicrobial agents,and help to assess the consequence of gut barrier damage and the efficacy of treatment.
3.Changes of circulating Tfr and Tfh cells in children with myasthenia gravis
Yajie CUI ; Fang CHEN ; Yanjun GUO ; Chunlan SONG
Chinese Journal of Microbiology and Immunology 2016;36(10):746-752
Objective To investigate the changes of follicular regulatory T cells ( Tfr cells) and follicular T helper cells ( Tfh cells) in peripheral blood of children with myasthenia gravis ( MG) . Methods We recruited 28 MG patients and 20 healthy subjects in this study. The percentages of Tfh and Tfr cells in peripheral blood samples were measured by flow cytometry. Real-time PCR was performed to detect the ex-pression of transcription factors and regulatory factors of Bcl-6, c-MAF, Blimp-1 and PD-1 at mRNA level. ELISA was used to detect the levels of IL-2, IL-6, IL-10 and IL-21 in plasma samples and the titers of Ach-Rab and PsMab. Results Compared with the healthy subjects, the MG patients showed higher percentages of Tfh cells and lower percentages of Tfr cells before receiving treatment. The expression of Bcl-6 and c-MAF on CD4+T lymphocytes cells at transcriptional level were significantly enhanced, while the expression of Blimp-1 on CD4+T cells and the expression of PD-1 on Treg cells at transcriptional level were inhibited in the MG patients in comparison with those in healthy subjects. Moreover, decreased levels of IL-2 and increased levels of IL-21 were found in plasma samples collected from the MG patients. Conclusion The decreased percentages of Tfr cells and increased percentages of Tfh cells in patients with MG resulted in abnormal ratios of Tfr/Tfh cells, which might be involved in the immunological pathogenesis of MG. Several changes in the patients with MG might be responsible for the imbalanced ratio of Tfr/Tfh cells, which included changes of IL-2 and IL-21 in microenvironment, enhanced expression of Bcl-6 and c-MAF at mRNA level and inhibited expression of Blimp-1 at mRNA level on CD4+T cells as well as over-expression of PD-1 at mRNA level on Treg cells.
4.Influence of Human Chorionic Gonadotropin on Testicular Germ Cell Apoptosis of Experimental Unilateral Cryptorchidism in Rats
guo-fang, WANG ; cui-ping, SONG ; yan-fang, REN ; guo-lin, ZHANG
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To explore the influence of human chorionic gonadotropin(HCG)on testicular germ cell apoptosis of experimental unilateral cryptorchidism in rats.Methods Forty immature male Sprague-Dawley rats were randomly divided into unilateral cryptorchi-dism group(n=20) and sham operation group(n=20).The 2 groups were divided into group treated with HCG and group without HCG.At age 21 days,unilateral cryptorchidism was produced.Half of the rats were injected with 20 U HCG from day 22 to 34 every other day.At age of 35 d and 60 d,rats were sacrificed for detection germ cell apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).Results Apoptosis index(AI) of cryptorchidism testis were higher compared with the scrotal testis in sham operation group(P0.05).AI of the scrotal testis of sham operation group and unilateral cryptorchidism group with HCG was higher compared with the correspond groups without HCG,and there were significant difference of AI between sham operation group and the correspond group without HCG at age 35 days(P0.05).Conclusions AI of testicular is increased both in cryptorchidism testis and scrotal testis in experimental unilateral cryptorchidism;HCG adds the number of apoptotic germ cells,and histology damage of testis is not completely recovery after stop using HCG.So clinical application of HCG must be cautious and operation ought to be done as early as possible in cryptorchidism.
5.Parameter optimal selection during pediatric 18F-FDG PET/CT examination
Fang GUO ; Jie CUI ; Hongyu ZHANG ; Zhongtai ZHANG ; Rongrong TIAN ; Ling YUAN
International Journal of Biomedical Engineering 2014;37(5):299-302
Objective Currently,pediatric 18F-FDG dose and acquisition durations are generally based on coarse extrapolation from adult guidelines.This study sought to determine whether shorter acquisition durations or a lower 18F-FDG injected activity could be used during pediatric 18F-FDG PET/CT examinations while maintaining diagnostic utility.Methods Thirty-six whole-body 18F-FDG PET/CT examinations were performed on 36 patients (weight,13-89 kg,(46.51±5.63) kg; age range,3-14 years old,(9.22±3.16) years old) with a weight-based injected activity (5.3 MBq/kg (0.144 mCi/kg)),fixed acquisition durations 180 S/FOV,VIP record acquisition mode using Discovery STE.For each examination,the Vip-mode data was truncated to form multiple datasets with shorter acquisition durations down to a minimum of 60 s/FOV (i.e.,60,80,100,120,140,160 s/FOV data were formed from single 180 s/FOV acquisition).168 image volumes were generated,randomized,and reviewed in a masked manner with corresponding CT image volumes by 6 radiologists.Overall,subjective adequacy and objective lesion detection accuracy by body region were evaluated.Results All examinations with maximum acquisition duration were graded as adequate and were used as the reference standard for detection accuracy.For patients more than 30 kg,when acquisition duration was more than 120 s/FOV,all PET/CT examinations were graded as adequate for clinical tasks,whereas,when acquisition duration was reduced to less than 120 s/FOV,lesion detection became less accurate.For patients less than 30 kg,lesion detection accuracy was perfect for acquisition times between 140 s/FOV and 180 s/FOV for all regions of the body.However,lesion detection became less accurate when imaging acquisition time was reduced less than 140 s/FOV.Conclusions When GE Discovery STE PET/CT was applied during pediatric PET/CT examination,using decreased acquisition times as a surrogate for 18F-FDG dose,18F-FDG dose can be reduced by approximately 33.33% when patients weigh over 30 kg were scanned for 180 s/FOV.For patients less than 30 kg,18F-FDG dose can be reduced by approximately 22.22% without losing diagnostic quality.Reduction of overall scan time potentially reduces motion artifacts,improves patient comfort,and decreases length of sedation.Alternatively,decreased 18F-FDG dose minimizes radiation risk.
6.Origin and Phylogenetic Characteristics of Dengue Virus Isolated from the Large Outbreak of Dengue in Guangdong Province in 2014
Qianfang GUO ; Guohui CUI ; Danyun FANG ; Huijun YAN ; Junmei ZHOU ; Lulu SI ; De WU ; Lifang JIANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):21-28
[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.
7.Hemodynamic changes vs pathologic foundation in rabbit models of radiation-induced lung injury: a preliminary study
Xiangming FANG ; Xiaoyun HU ; Chunhong HU ; Hongwei CHEN ; Lei CUI ; Qinghua ZHANG ; Jian GUO ; Juying ZHOU
Chinese Journal of Radiology 2012;46(2):158-163
Objective To study the hemodynamic changes and pathologic foundation of rabbit models of radiation-induced lung injury (RILI) via 64-slice CT pulmonary perfusion imaging ( CTPI),in order to seek the correlation between the alterations of the hemodynamic parameters and pathophysiology.Methods Seventy-two healthy New Zealand rabbits were randomly classified into two groups:test group ( n =36),received 25 Gy with single fraction irradiation in a whole unilateral lung; control group ( n =36),received sham-irradiation.Each group was divided into 12 subgroups respectively according to post- and pseudo-irradiation time points (1,6,12,24,48,72 h and 1,2,4,8,16,24 w).Each rabbit underwent HRCT and CTPI at every pre- and post-radiation time point.All rabbits were sacrificed,and morphology of specimens was observed using light- and electron microscope. The changing regularity of HRCT,CTPI parameters and pathology were analyzed and compared with each other in order to find the correlation among them.The CTPI parameters of the test and control groups were compared using t test.The CTPI parameters and pathological values were analyzed using linear correlation with two variables,the detection rates of RILI by CTPI and HRCT was compared using Chi-square test.Results ( 1 ) The changes of CTPI parameters from control group after irradiation was relatively stable,but in test group those parameters including rBF,rBV and rPS,at pre- and post-irradiation time points (0,72 h and 2 w),were respectively 1.01 ± 0.09,1.86 ± 0.20,1.43 ±0.12,1.03 ±0.08,1.63 ±0.19,1.56±0.14,0.96±0.12,1.54 ±0.17 and 1.83 ±0.24.The corresponding parameters before and after irradiation were significantly different ( t =2.90-6.37,P > 0.05).(2)In test group,capillary endothelial cells,basement membrane and alveolar epithelial cells,as the main injury targets,showed certain alterations in pathology.There was a significant correlation between the changes of CTPI parameters ( rBF and rBV) and pathophysiology in control group ( r =0.74,0.83,P <0.05 ),with the dependent relationship between rPS and the amounts of RBC outside the capillary and the destruction of basement membrane( r =0.87,0.88,P < 0.01 ).(3)The detection rate of RILI with CTPI (72.2%,26/36) was obviously higher than that with HRCT( 16.7%,6/36,x2 =4.37,P =0.036).Conclusions CTPI parameters is capable of revealing the rule of hemodynamic process and reflecting the pathophysiologic state of different stages of RILI.By the time of detecting RILI,the detection rate of CTPI is clearly superior to that of HRCT,which yields potential value in predicting RILL
8.Metabolic characteristics and risk factors for newly diagnosed type 2 diabetes mellitus combined with non-alcoholic fatty liver disease
Dongxia FU ; Haibin CUI ; Jinxiu XU ; Guangya WANG ; Fang GAO ; Ningning GUO
Clinical Medicine of China 2013;(3):237-240
Objective To explore metabolic characteristics of and risk factors for newly diagnosed type 2 diabetes mellitus(T2DM) combined with non-alcoholic fatty liver disease (NAFLD).Methods One hundred and forty-two cases of newly diagnosed T2DM were divided into two groups according to whether they have comorbid NAFLD:group A (without NAFLD,n =79) and group B (combined with NAFLD,n =63).Data collected included body height,body weight,blood pressure,fasting plasma glucose (FPG),blood lipid,serum uric acid (UA),HbA1c and fasting insulin,body mass index and insulin resistance index with homeostasis model(HOMA-IR) were calculated to compare the clinical and biochemical parameters between groups A and B.Results (1) The difference of age and blood pressure between groups A and B were not statistical different (P > 0.05).Compared with group A,BMI ((26.79 ± 1.93) kg/m2 vs (24.61 ± 2.46) kg/m2,t =5.76),FINS((15.49±2.44) mU/L vs (13.20±2.17) mU/L),t =5.91),HOMA-IR((6.74± 1.32) vs (5.65 ±1.10),t =5.37),glycerin trimyristate (TG) ((2.94 ± 0.65) mmol/L vs (1.74 ± 0.46) mmol/L),t =12.86),low density lipoprotein cholesterin (LDL-C) ((3.46 ±0.73) mmol/L vs (2.78 ±0.86) mmol/L,t =5.07) and UA((342.41 ±71.49) mmol/L vs (312.98 ±66.24) mmol/L,t =2.54) were significantly higherand hight density lipoprotein cholesterin (HDL-C) ((0.99 ± 0.17) mmol/L vs (1.21 ± 0.29) mmol/L,t =5.33) was significantly lower in group B (P < 0.05).(2) Using whether to combined with NAFLD as dependent variable,and BMI,FINS,HOMA-IR,TG,LDL-C,HDL-C and UA as independent variable,logistics regression analysis showed that BMI,HOMA-IR and TG were risk factors for NAFLD(OR =2.838,19.241,and 2.019 respectively,P < 0.05).Conclusion Newly diagnosed type 2 diabetes mellitus combined with NAFLD have more obvious dyslipidemia and insulin resistance.Obesity,insulin resistance,hyper-triglyceridemia are risk factors for newly diagnosed type 2 diabetes mellitus combined with NAFLD.
9.Correlation between polymorphism of cytochrome P450 1A2 gene and antidepressant clinical efficacy of duloxetine
Dongbo LIU ; Shuying LI ; He CUI ; Yali WANG ; Binhua ZHU ; Fang CHEN ; Huirong GUO
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(2):144-147
Objective To investigate the relationship between C734A and G-2964A polymorphism of cytochrome P450 1A2 gene and clinical efficacy of duloxetine.Methods 223 patients with depression were treated with duloxetine for six weeks.The clinical efficacy was evaluated with the Hamilton rating scale for depression (HAMD) ;single nucleotide polymorphism (SNP) at position C734A and G-2964A of CYP1A2 gene were identified with restriction fragment length polymorphism(RFLPs) ;then one-way ANOVA was adopted to analyze the relationship between SNP and clinical efficacy.Results (1) In 223 patients,the frequency of allele A at locus 734 was 63.64%,while that of allele A at locus-2964 was 26.82%.(2) 220 patient underwent the whole treating course.The conjoint analysis of two locuses indicated that the decreasing ratio of HAMD score of high-activity group,middle-activity group and low-activity after treatment was (56.05± 10.13) %,(66.36± 8.66) % and (73.82± 7.10) % respectively,the differences obtained by pairwise comparison of the three groups were of statistical significance(P<0.01).Conclusion There is close relationship between C734A and G-2964A polymorphism of CYP1A2 gene and clinical efficacy of duloxetine in the treatment for depression.
10.Detection of Aspergillus fumigatus genome load in human whole blood samples by real-time quantitative polymerase chain reaction and its clinical application
Xizeng CUI ; Enling MA ; Junren KANG ; Guangliang GUO ; Jiabin FANG ; Yunfei XU
Chinese Journal of Clinical Nutrition 2011;19(3):154-158
Objective To establish a real-time quantitative polymerase chain reaction (RQ-PCR) assay for fast detection of Aspergillus fumigatus genome in human whole blood samples and explore its clinical application.Methods The primers and the TaqMan-probe were designed on the basis of the multi-copy ITS1-5. 8S region of the rDNA of Aspergillus fumigatus. The Aspergillus fumigatus genomic DNA were extracted with QIAamp(R) DNA Blood Mini Kit.A 20 μl RQ-PCR amplification system was established, and the simulated blood samples containing various given load of Aspergillus fumigatus genome and the 66 whole blood samples of the surgical febrile patients were examined. Results The detection limit of the RQ-PCR instrument is 10-1 genomes/μl DNA sample,namely 78 CFU/ml whole blood. The specificity and the sensitivity were 94. 25% and 99. 04% respectively; and the positive predictive value and negative predictive value were 97. 63% and 97. 62% respectively. The average relative error of the quantitative results was (3. 67 ±13. 19)%, and the intra- and the inter-assay average coefficients of variation were (12.38 ± 1. 53)% and (16. 27 ±2. 72)% , respectively. The average recovery rate of Aspergillus fumigatus genomic DNA in human whole blood samples was (107. 81 ±25. 92)% , and the average coefficient of variation of the average recovery rate was (26. 24 ± 5.62) % . No Aspergillus fumigatus genomic DNA was detected among the 66 blood samples of the surgical febrile patients. Conclusions The RQ-PCR assay for fast quantitative detection of Aspergillus fumigatus genome in human whole blood samples is of high sensitivity, specificity,accuracy and precision. The Aspergillus fumigatus genome was not detected in this group of surgical febrile patients.