1.Application and evaluation of a probe melting analysis-based assay for detection of Isoniazid-resistant mutations in Mycobacterium tuberculosis
Siyu HU ; Jianjun NIU ; Shengmao QUAN ; Jianwei HUANG ; Qingge LI
Chinese Journal of Laboratory Medicine 2011;34(10):935-939
Objective To validate the performance of a PMA-based assay for detection of INH-resistant mutations in MTB and investigate the mutation characteristic of INH-resistance.Methods The MTB standard strain H37Rv was from National Tuberculosis Reference Laboratory,1 wild-type strain and 1 katG S315T ACC mutant strain were from Xiamen CDC,7 MTB INH-resistant strains with known INH resistance mutations were from Shenzhen Center for Chronic Disease Control,Henan CDC,No.309 Hospital of PIA and Xiamen CDC.707 MTB clinical isolates were from Xiamen CDC,Xiamen No.1 Hospital and Zhangzhou CDC,126 sputum samples were from Xiamen Tongan CDC.The genomic DNA of the MTB standard strain H37Rv,7 MTB INH-resistant strains and 833 clinical samples,were extracted with Xiamen Zeesan Mycobacterium tuberculosis Isoniazid-resistance Mutation Test Kit using the thermal lysis method.The genomic DNA of 1 wild-type strain and 1 katG S315T ACC mutant strain were extracted with AxyPrep Bacterial Genomic DNA Miniprep Kit.The mutations were discriminated by the △Tm between the samples and wild type control in katG315 codon,inhA promoter -17 to -8 region,ahpC promoter region -44 to -30 and - 15 to 3,and inhA94 codon.A 10-fold dilution series of MTB DNA from 3 × 105 to 300 copies/ reaction obtained from a wild-type strain and a katG S315T ACC mutant strain,respectively,were prepared to determine the analytical sensitivity.Seven MTB INH-resistant strains with 9 predetermined mutations were used for the analytical specificity assay,and 5 mutants of which were used for the repeatability assay.The clinical detection performance of PMA assay were confirmed by the sequencing method in 833 samples.Results Results could be obtained within 3 hours from DNA extraction to PMA assay,including 46 samples in a standard 96-well real-time PCR instrument simultaneously.The analytical sensitivities of PMA were 300 copies/reaction for both the wild-type strain and katG S315T ACC mutant strain.Nine INH-resistant point mutations could be discriminated and 5 of which had standard deviations of melting temperature less than 0.5 ℃.Fully concordant results of mutant locus between PMA assay and sequencing were obtained in all 162 mutant samples.INH-resistant mutations in the four loci were found in 19.4% (162/833) samples by PMA assay in Xiamen and Zhangzhou.Among the 14 lNH-resistant mutant types detected,katG S315T ( AGC→ACC),inhA promoter - 15C→T and katG S315N (AGC→AAC) accounted for 83.3% (135/162) of the overall mutations.
2.Expression levels and targets of more than fifty miRNAs in glioma
Taiqi LIU ; Siyu CHEN ; Wende LI ; Ren HUANG
Chinese Journal of Comparative Medicine 2016;26(7):79-84
miRNAs are a class of small endogenous RNAs that degrade target mRNAs or repress their translation process. Several miRNAs in glioma are up?regulated, while some others down?regulated. Some miRNAs promote tumorigenesis; some others, however, play a similar function of tumor suppressor genes. Therefore, studies on the expression profiles of miRNAs in glioma may afford auxiliary basis for early clinical diagnosis and novel srtategies for therapy of glioma. This paper will review on researches about the expression levels of miRNAs and their targets in glioma.
3.Analysis of governmental responsibilities in health protection as inspired by the model of Sanming's healthcare reform
Siyu TAO ; Hong LE ; A'hong HUANG ; Pengqian FANG ; Fang WANG ;
Chinese Journal of Hospital Administration 2017;33(4):252-254
Success of Sanming's heahhcare reform relies heavily on the government's dominant role.Based on this inspiration,the paper discussed the responsibilities expected of the government in health protection of the people as required by national strategy,public interests and market demands,and presented the corresponding responsibilities of the government in the protection,leadership,scrutiny and management for the sake of public health.In addition the paper recommended on policies to make up for the absence and dislocation of governmental responsibilities,including all-out efforts for legislations in health protection,greater government subsidy for health protection,development of a comprehensive supervision system,and establishment of entities accountable for people's health in general.
4.Effect of selection of statistical uncertainty of control points in Monaco planning system on dose calculation in nasopharyngeal carcinoma
Siyu WU ; Xiaoyan HUANG ; Wufei CAO ; Li CHEN
Chinese Journal of Radiation Oncology 2021;30(2):170-174
Objective:To explore the influence of the selection of statistical uncertainty of control points in Monaco planning system on the dose distribution of nasopharyngeal carcinoma (NPC), aiming to provide the statistical uncertainty of single control point in Monte Carlo calculation which satisfies clinical needs.Methods:First, nine 10 cm×10 cm square fields with an equal interval of gantry angle were designed and five cases of 9-field intensity-modulated radiotherapy (IMRT) and five cases of single-arc volumetric-modulated arc therapy (VMAT) plans were randomly selected, Then, quality assurance (QA) verification plan using patient CT as QA phantom was created. Second, the grid spacing was selected as 3 mm during the calculation of dose distribution of QA plan. The statistical uncertainties of single control point were selected as 1%, 2%, 3%, 4% and 5%, respectively. Last, the deviation of dose distribution between different statistical uncertainties and 1% statistical uncertainty was analyzed.Results:For a square field and single IMRT field, the dose deviation of center point was almost 7% while the statistical uncertainty was selected 4%. For 9-field IMRT and single-arc VMAT, the dose deviation of center point was ≤ 1.5% and the average dose deviation of PTV was ≤ 0.3% when the statistical uncertainty of control points was changed from 1% to 5%. The percentage of the point dose deviation of the coronary plane of ≤ 1% was greater than 99% when the statistical uncertainty was ≤ 3% for 9-filed IMRT and 4% for single-arc VMAT.Conclusions:For the Monaco treatment planning system based on Monte Carlo calculation, the changes in the statistical uncertainty of control point from 1% to 5% exert significant effect upon the single field. In clinical application, the statistical uncertainty of control point should be ≤ 3% for 9-field IMRT and ≤ 4% for single-arc VMAT.
5.“Happy Reading Club” established by undergraduates and its problems
Qing HOU ; Xiaoyan Lü ; Jiongjiong HUANG ; Wei JI ; Sheng YU ; Siyu ZHANG
Chinese Journal of Medical Library and Information Science 2014;(10):44-46
Establishment of the “Happy Reading Club” was to create a personal reading space for undergraduates. The reading situation of undergraduates was analyzed with the creation processes of “Happy Reading Club” summa-rized.The reading popularizing activities were described, including questionnaire investigation of reading, sign of DIY “tree”, and recommendation of classic booklist, with the existing problems pointed out.
6.Establishment and application of a tandem affinity purification system of innate immune regulatory protein PKR
Yuye LI ; Zhaoduan LIANG ; Siyu WU ; Jiong XIE ; Junfang HE ; Minhao WU ; Xi HUANG ; Ping ZHANG
Chinese Journal of Microbiology and Immunology 2011;31(6):487-491
Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.
7.Clinical Study of 5% D-fructose Injectio for Energy Supply in Surgery Patients
Desheng MENG ; Liang CHEN ; Qunyou TAN ; Jian HUANG ; Siyu WU ; Taiqian GONG ; Wei WU ; Yaoguang JIANG
China Pharmacy 2001;0(11):-
OBJECTIVE: To observe the clinical effect of 5% D-fructose injectio on energy supply in surgery patients. METHODS: By setting 5% glucose injectio as control,the influence of 5% D- fructose injectio on blood sugar level,liver and kidney function indices was detected.RESULTS: 5% D-fructose injectio did not influence liver and kidney functions, serum uric acid and RESULTS: of routine examination of blood and urine.Compared with control group, the change of blood sugar level in experiment group was slighter.CONCLUSION: 5% D-fructose injectio is effective and safe for energy supply in surgery patients.
8.PROGNOSTIC SIGNIFICANCE OF N1 STAGE IN NON-SMALL CELL LUNG CANCER
Siyu WANG ; Yilong WU ; Zhifen HUANG ; Wei QU ; Xiening YANG ; Hui YU
Tumor 2001;(2):117-119
Objective To investigate the prognostic significance of N1 stage in non-small cell lung cancer (NSCLC) by comparing is survival with N0 and N2 stages. Methods From Jan. 1982 to Dec. 1994, 138 NSCLC patients with surgical-pathological N1 stage had received complete mediastinal lymph node dissection. Two subgroups of N1 node were identified by using New Regional Lymph Node Classification for Lung Cancer Staging. Their prognostic significances were tested and 5-year survival rates were compared with those of surgical-pathological N0 (307 cases) and N2 (176 cases) patients received radical operation during the same period. Results The overall 5-year suvival rate of N1 patients was 30.4 %. five-year survival was significantly better when N1 involvement was intralobar as compared with extralobar involvement, 50.3 % versus 26.5 % (P=0.001). Intralobar N1 5-year survival was similar to that of N0 (51.4 % vs 50.3 %), and extralobar N1 5-year survival was similar to that of N2 with singl focus (26.5 % vs 23.5 %). Conclusion N1 stage in NSCLC is a compound with two subgroups; the prognostic significance of intralobar subgroup is related to N0 stage and extralobar subgroup is related to that of single focus N2.
9.The dignostic value of serum vitamin B12 in predicting rebleeding in patients with liver cirrhosis of esophagogastric varices
Siyu JIANG ; Qiuyan LIN ; Yingjie AI ; Ling WU ; Xiaoquan HUANG ; Shiyao CHEN
Journal of Chinese Physician 2021;23(3):343-346
Objective:The aim of this study was to investigate the prognostic value of vitamin B12 as the non-invasive biomarker to predict long-term rebleeding rate in cirrhotic patients with esophagogastric varices.Methods:From Dec 1, 2016 to Dec 31, 2017, cirrhotic patients with esophagogastric varices who had been admitted to Zhongshan Hospital affiliated to Fudan University were enrolled. All these patients received endoscopic treatment to prevent variceal rebleeding. The serum vitamin B12 and folic acid levels were measured in all of them. The receiver operating characteristic (ROC) analysis, Kaplan-Meier analysis, univariate and multivariate cox regression analysis were conducted to explore the value of vitamin B12 in predicting 3-year variceal rebleeding in cirrhotic patients with esophagogastric varices after endoscopic treatment.Results:115 patients were included. The ROC curve analysis indicated that the optimal cutoff value of vitamin B12 for 3-year variceal rebleeding was 567.25 pg/ml. According to the cut-off value, the patients were divided into high-level vitamin B12 group ( n=49) and low-level vitamin B12 group ( n=66). Compared with the low vitamin B12 group, the high vitamin B12 group had lower albumin level, less male (63.3% vs 80.3%), and higher 3-year rebleeding rate ( P<0.05). Cox analysis showed that vitamin B12 and platelet were independent prognostic factors for 3-year rebleeding in patients with variceal bleeding. Conclusions:Elevated peripheral blood vitamin B12 predicts a higher risk of long-term rebleeding in patients with liver cirrhosis and esophagogastric varices.
10.Change of serum myeloperoxidase and lipoxin A4 level in coronary heart disease patients with anxiety and/or depression.
Siyu LIANG ; Xiangping LI ; Wenyu HUANG ; Hairong GONG
Journal of Central South University(Medical Sciences) 2013;38(4):370-375
OBJECTIVE:
To investigate the change of serum myeloperoxidase (MPO) and lipoxin A4 (LXA4) in coronary heart disease (CHD) patients with anxiety and depression and its clinical significance.
METHODS:
From December 2010 to February 2011, 143 CHD patients and 44 non-CHD patients (the control group) hospitalized in the Department of Cardiology at the Second Xiangya Hospital were enrolled. The hospital anxiety and depression scale (HADS) was used to evaluate the psychological state of all patients and the CHD patients were assigned to an anxiety and depression group (n=57) or a non-depression and anxiety group (n=86). The serum levels of high sensitive C-reactive protein (Hs-CRP), MPO, and LXA4 were examined, and the ratio of MPO and LXA4 (M/L) was calculated.
RESULTS:
The levels of Hs-CRP, MPO, and LXA4 as well as M/L ratios in both CHD groups were significantly higher than those in the control group (all P<0.01). Compared with the non-anxiety and depression group, the levels of MPO and LXA4, and M/L ratios in the anxiety and depression group increased (all P<0.05). Correlation analysis showed that MPO was positively correlated with the score of HADS-total (HADS-t), HADS-anxiety (HADS-a), or HADS-depression (HADS-d), while LXA4 was negatively correlated with HADS-t or HADS-d. Multiple ordinal logistic regression analysis revealed that higher HADS-t score, stable angina, unstable angina, and acute myocardial infarction were the independent impact factors for the elevation of M/L ratio.
CONCLUSION
Anxiety and depression may aggravate the inflammatory response in CHD patients. The imbalance between inflammation and anti-inflammation may be part of the mechanism.
Aged
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Anxiety
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complications
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Case-Control Studies
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Coronary Disease
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blood
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complications
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Depression
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complications
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Female
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Humans
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Inflammation
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Lipoxins
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blood
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Male
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Middle Aged
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Peroxidase
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blood