1.Development of a droplet digital polymerase chain reaction assay for the sensitive detection of total and integrated HIV-1 DNA
Lin YUAN ; Zhiying LIU ; Xin ZHANG ; Feili WEI ; Shan GUO ; Na GUO ; Lifeng LIU ; Zhenglai MA ; Yunxia JI ; Rui WANG ; Xiaofan LU ; Zhen LI ; Wei XIA ; Hao WU ; Tong ZHANG ; Bin SU
Chinese Medical Journal 2024;137(6):729-736
Background::Total human immunodeficiency virus (HIV) DNA and integrated HIV DNA are widely used markers of HIV persistence. Droplet digital polymerase chain reaction (ddPCR) can be used for absolute quantification without needing a standard curve. Here, we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods::The limit of detection, dynamic ranges, sensitivity, and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat (LTR) and human CD3 gene (for total HIV DNA) and ACH-2 cells (for integrated HIV DNA). Forty-two cases on stable suppressive antiretroviral therapy (ART) were assayed in total HIV DNA and integrated HIV DNA. Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive (CD4 +) T-cell counts, CD8 + T-cell counts and CD4/CD8 T-cell ratio, respectively. The assay linear dynamic range and lower limit of detection (LLOD) were also assessed. Results::The assay could detect the presence of HIV-1 copies 100% at concentrations of 6.3 copies/reaction, and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction (95% confidence intervals [CI]: 3.6-6.5 copies/reaction) with linearity over a 5-log 10-unit range in total HIV DNA assay. For the integrated HIV DNA assay, the LLOD was 8.0 copies/reaction (95% CI: 5.8-16.6 copies/reaction) with linearity over a 3-log 10-unit range. Total HIV DNA in CD4 + T cells was positively associated with integrated HIV DNA ( r = 0.76, P <0.0001). Meanwhile, both total HIV DNA and integrated HIV DNA in CD4 + T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8 + T-cell counts. Conclusions::This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity. It can be readily adapted for measuring HIV DNA with non-B clades, and it could be beneficial for testing in clinical trials.
2.Study on causal relationship between walking pace with interverbral disc degeneration and low back pain based on two-sample bidirectional Mendelian randomisation
Chao SU ; Yuxiao TIAN ; Lifeng ZHUANG ; Di XIA ; Manhong YANG ; Tianhao WAN ; Qing ZHANG
Chongqing Medicine 2024;53(20):3113-3119
Objective To investigate the causal relationship between walking pace(WP)with interver-tebral disc degeneration(IVDD)and low back pain(LBP)by using two-sample Mendelian randomisation(MR)analysis method.Methods A genome-wide association study database of WP,IVDD and LBP was ob-tained,and single nucleotide polymorphisms(SNP)strongly associated with exposure were obtained as the in-strumental variables.The two confounders of smoking and sedentary were conservatively removed,and the MR analyses were performed by inverse variance weighting(IVW),weighted median,and MR-Egger methods to assess the causal relationship between WP with IVDD and LBP.The consistency and accuracy of results were ensured by heterogeneity,diversity tests,MR-PRESSO and negative control.Results The I VW results showed a significant negative correlation between WP and IVDD(OR=0.348,95%CI:0.183-0.661,P=0.001)and vice versa(OR=0.972,95%CI:0.953-0.992,P=0.007).Meanwhile there was also a significant negative correlation between WP and LBP(OR=0.214,95%CI:0.119-0.385,P<0.001),but no causal re-lationship was found between LBP and WP.Conclusion Faster WP could effectively prevent the occurrence of IVDD and LBP,and IVDD may contribute to the occurrence of slow walking.
3.Automatic delineation of rectal cancer target volume and organs at risk based on convolutional neural network
Xiang XIA ; Jiazhou WANG ; Lifeng YANG ; Zhen ZHANG ; Weigang HU
Chinese Journal of Radiation Oncology 2020;29(5):374-377
Objective:To realize automatic delineation of rectal cancer target volume and normal tissues and improve clinical work efficiency.Methods:The deep learning method based on convolutional neural network was adopted to construct neural network, learn and realize automatic delineation, and compare the differences between automatic delineation and manual delineation.Results:Two hundred and ten cases with rectal cancer were randomly assigned to a training set of 190 and a validation set of 20. The complete delineation of a single case took about 10s; the average Dice of CTV was 0.87±0.04; the average Dice of other normal tissues was bigger than 0.8; the Hausdorff distance (HD) index of CTV was 25.33±16.05; the mean distance to agreement (MDA) index was 3.07±1.49, and the Jaccard similarity coefficient (JSC) index was 0.77±0.07.Conclusion:The deep learning method based on full convolutional neural network can realize the automatic delineation of rectal cancer target volume and improve work efficiency.
4.A phase Ⅲ clinical trial study on the safety and immunogenicity of ACYW135 group meningococcal conjugate vaccine inoculated in 3 month old infants
Zhiqiang XIE ; Dongyang ZHAO ; Haitao HUANG ; Jinbo GOU ; Wei ZHANG ; Yongli YANG ; Lili HUANG ; Yanxia WANG ; Xue WANG ; Lifeng XU ; Tao ZHU ; Shengli XIA
Chinese Journal of Preventive Medicine 2020;54(9):947-952
The aim of this study was to evaluate the safety and immunogenicity of the first domestic ACYW135 meningococcal conjugate vaccine and a control vaccine named AC group meningococcal conjugate vaccine for 3 months (90-119 days) infants. From February 2017 to June 2018, a randomized, blinded, and similar vaccine-controlled clinical trial design was adopted at the Henan Vaccine Clinical Research Base. The subjects were 3 months old healthy infants, a total of 720, based on a 1∶1 ratio. The random allocation table for entry was randomly assigned to the experimental group and the control group. According to the 3, 4, and 5 month-old vaccination procedures, the subjects were vaccinated with test vaccine (ACYW135 group meningococcal conjugate vaccine) and control vaccine (group A group C meningococcal polysaccharide conjugate vaccine), of which 720 were given the first dose, 696 were given the second dose (test group: 346; control group: 350), and 692 were given the third dose (test group: 344; Control group: 348). The overall adverse reaction rate of the test vaccine was 21.90% (230 cases), which was lower than the 32.04% (339 cases) of the control vaccine (<0.001). The incidence of systemic adverse reactions was 19.52% (205 cases), which was lower than that of the control vaccine (27.69%) (293 cases) (<0.001). The local adverse reaction rate was 3.04% (32 cases), which was lower than the control group (7.84%) (83 cases) (<0.001). The graded adverse reaction test vaccine was 0.57% (6 cases), which was lower than the control group of 2.36% (25 cases) (<0.001). The positive conversion rate of anti-bacterial serum antibodies showed that there was no significant difference between the test vaccine group A (91.42%), C (88.76%) and the control vaccine (92.92%) (87.02%) (>0.05). Group Y and W135 was 88.17% (298 cases), 99.41% (336 cases), respectively. The GMT results showed that the test vaccine group A was 56.24, the control vaccine was 57.43 (>0.05); the group C test vaccine (43.53) was higher than the control group (27.28) (<0.001). The group Y and W135 are 89.22 and 140.66, respectively. Among them, the proportion of the group C GMT antibody ≥ 1∶128 for test vaccine (31.07%, 105 cases) was higher than the control vaccine (16.22%, 55 cases) (<0.001). ACYW135 group meningococcal conjugate vaccine has more safety and immunogenicity after application to 3 month old infants.
Antibodies, Bacterial
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Humans
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Infant
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Meningococcal Vaccines
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adverse effects
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immunology
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Vaccines, Conjugate
5.The early evaluation of lowGdose CT perfusion imaging for the treatment efficacy of lung cancer with anrotinib hydrochloride
Lijing FAN ; Xuejun CHEN ; You YUN ; Xiaoxian ZHANG ; Lifeng WANG ; Xiaoyan WEI ; Weili XIA ; Hailiang LI
Journal of Practical Radiology 2019;35(6):977-980
Objective To explore whether CT perfusion imaging (CTPI)parameters can early predict the curative effect of anlotinib hydrochloride and their predictive accuracy for the treatment in lung cancer patients.Methods 2 6 patients with advanced nonGsmall cell lung cancer (NSCLC)were treated with anlotinib hydrochloride and underwent CTPI scanning before chemotherapy,after the first and second treatment cycle respectively.The average values of perfusion value (PV),peak enhancement image (PEI),time to peak (TTP),blood volume (BV)and the change rate of these parameters after one treatment cycle every time were measured and recorded. According to the response evaluation criteria in solid tumors 1.1 (RECIST1.1),the maximum diameter of the target tumor was measured and the tumor regression rate after two treatment cycles was calculated.Then a correlation analysis was conducted between the change rate of perfusion parameters (PV%,PEI%,TTP%,BV%)after one treatment cycle and the tumor regression rate (D%)after two treatment cycles. The ROC curve was performed to evaluate the accuracy of those parameters.Results PV after one treatment cycle was significantly lower than that before treatment,and PV% showed a statistical difference (P=0.00).The PV% after one treatment cycle was positively correlated with D% after two treatment cycles (r=0.56).In addition,the AUC of PV% and BV% were 0.99 and 0.88 respectively, and specificity were both 100%,with sensitivity respectively 75.7% and 82.6%.Conclusion CTPI can early reflect the curative effect of anlotinib hydrochloride for advanced NSCLC and provide more options for clinical evaluation.
6. Cancer incidence and mortality in Pearl River Delta Area, 2009-2013
Liang XIA ; Yanjun XU ; Xiaojun XU ; Ruilin MENG ; Lifeng LIN
Chinese Journal of Oncology 2019;41(5):393-397
Objective:
To estimate the incidence, mortality and characteristics of cancer in Pearl River Delta Area of Guangdong Province between 2009-2013.
Methods:
Based on five population-based cancer registration data from Guangzhou, Shenzhen, Zhongshan, Jiangmen and Sihui spanning from 2009 to 2013, along with those corresponding population data, the incidence and mortality rates were estimated by gender and age groups. Chinese standard population derived from the 2000 Population Census and Segi′s standard population were used for age-standardized incidence and mortality rates.
Results:
Between 2009 and 2013, the crude cancer incidence rate was 262.50/100 000, 274.76/100 000 in male and 249.49/100 000 in female. After adjusting for Chinese and Segi′s standard population, the age-standardized incidence rates were 225.63/100 000 and 219.88/100 000, respectively. The crude mortality rate was 175.51/100 000, 222.92/100 000 in male and 127.46/100 000 in female, respectively. After adjusting for Chinese and Segi′s standard population, the age-standardized mortality rates were 116.02 /100 000 and 114.31/100 000, respectively. The incidence rates were at low levels in the population less than 40 years old, thereafter went up rapidly with age especially in male, and then reached the peak in the population aged 80 and above. As with incidence, the mortality rates kept at low levels in the population before their 50 s and then rose up steadily with age until peaking in the 85+ age group. The most common cancers were female breast cancer, lung cancer, colorectal cancer, liver cancer and nasopharyngeal cancer with descending incidence rate. Lung cancer, liver cancer, colorectal cancer, female breast cancer and nasopharyngeal cancer were the top five cancer-attributable causes of death.
Conclusions
Currently, Pearl River Delta Area were faced with huge cancer burden. Lung cancer, colorectal cancer, nasopharyngeal cancer, female breast cancer and male liver cancer are predominant cancers and more efforts should be made to fight against them.
7. Clinical value of quantitative detection of DNA aneuploidy in the diagnosis and treatment of cervical lesions in middle-aged and senile women
Lifeng WANG ; Yunheng ZHU ; Xiuxiang ZHU ; Ziyin XIA
Chinese Journal of Geriatrics 2019;38(11):1285-1288
Objective:
To investigate the clinical value of quantitative detection of DNA aneuploidy in the diagnosis and treatment of cervical lesions in middle-aged and senile women.
Methods:
A total of 1 404 middle-aged and elderly women who underwent screening for early cervical lesions were retrospectively studied.Patients were divided into the two groups: the 40-49 years old group(n=897)and the 50-78 years old group(n=507). Cervical lesions were screened by DNA ploidy analysis and the results were compared with those screened by liquid-based cytology, colposcopy and high-risk human papillomavirus(HPV).
Results:
The positive detection rate of HPV by DNA ploidy analysis was 54.4%(764/1 404). Of 1 404 patients, HPV16/18 infection accounted for 21.3%(299/1 404). The detection rate of heteroploid cells was 50.92%(715/1 404). There was a significant positive correlation between HPV infection type and cervical epithelial cell ploidy changes(
8. Treatment and improvement of scar hyperplasia by panax notoginseng based on network pharmacology study on molecular mechanisms
Jingwen HUANG ; Xue HAN ; Xu ZHAO ; Xubo LI ; Xingzhong DONG ; Lifeng AN ; Shunxin JI ; Xianghe XIA
International Journal of Traditional Chinese Medicine 2019;41(12):1353-1359
Objective:
We predicted the molecular mechanism of Panax notoginseng in the treatment and improvement of scar hyperplasia, by using the methods of network pharmacology and bioinformatics.
Methods:
We collected of related active constituents and targets of Panax notoginseng by retrieving TCM systems pharmacology database and analysis platform (TCMSP), and collected of related active constituents and targets of scar by Genecards database and OMIM database. Cytoscape 3.6.1 software was used to construct "drugs-chemical components-targets-diseases" interaction network diagram. The protein in teraction network map (PPI) was constructed by STRING database. The key targets were used to analyze gene ontology (GO) enrichment and kyoto encyclopedia of genes and genome (KEGG) pathway enrichment.
Results:
Totally 7 chemical compponents, including beta-sitosterol, quercetin, Stigmasterol and etc. and 108 targets, including AKT1, JUN, MAPK1, IL6 and ect. Panax notoginseng exerts its effects on scar mainly by acting on signal pathways, including PI3K-AKt signal pathway, MAPK signal pathway, TNF signal pathway and ect.
Conclusions
Based on the methodology of network pharmacology, this study preliminarily predicted the major targets and pathways Panax notoginseng in the treatment of scar, providing a direction for further studies.
9.Cancer incidence and mortality in Pearl River Delta Area, 2009?2013
Liang XIA ; Yanjun XU ; Xiaojun XU ; Ruilin MENG ; Lifeng LIN
Chinese Journal of Oncology 2019;41(5):393-397
Objective To estimate the incidence, mortality and characteristics of cancer in Pearl River Delta Area of Guangdong Province between 2009?2013. Methods Based on five population?based cancer registration data from Guangzhou, Shenzhen, Zhongshan, Jiangmen and Sihui spanning from 2009 to 2013, along with those corresponding population data, the incidence and mortality rates were estimated by gender and age groups. Chinese standard population derived from the 2000 Population Census and Segi′s standard population were used for age?standardized incidence and mortality rates. Results Between 2009 and 2013, the crude cancer incidence rate was 262.50/100 000, 274.76/100 000 in male and 249.49/100 000 in female. After adjusting for Chinese and Segi′s standard population, the age?standardized incidence rates were 225.63/100 000 and 219.88/100 000, respectively. The crude mortality rate was 175.51/100 000, 222.92/100 000 in male and 127.46/100 000 in female, respectively. After adjusting for Chinese and Segi′s standard population, the age?standardized mortality rates were 116.02 /100 000 and 114.31/100 000, respectively. The incidence rates were at low levels in the population less than 40 years old, thereafter went up rapidly with age especially in male, and then reached the peak in the population aged 80 and above. As with incidence, the mortality rates kept at low levels in the population before their 50 s and then rose up steadily with age until peaking in the 85+ age group. The most common cancers were female breast cancer, lung cancer, colorectal cancer, liver cancer and nasopharyngeal cancer with descending incidence rate. Lung cancer, liver cancer, colorectal cancer, female breast cancer and nasopharyngeal cancer were the top five cancer?attributable causes of death. Conclusions Currently, Pearl River Delta Area were faced with huge cancer burden. Lung cancer, colorectal cancer, nasopharyngeal cancer, female breast cancer and male liver cancer are predominant cancers and more efforts should be made to fight against them.

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