1.Development of an Integrated Disposable Device for SARS-CoV-2 Nucleic Acid Extraction and Detection
Ma JING ; Hao YANZHE ; Hou MEILING ; Zhang XIAOSHAN ; Liu JINGDUAN ; Meng HAODI ; Chang JIANGBO ; Ma XUEJUN ; Liu JIHUA ; Ying QINGJIE ; Wang XIANHUA ; Li HONGXIA ; Cao YUXI ; Zhang XIAOGUANG
Biomedical and Environmental Sciences 2024;37(6):639-646
Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Methods We designed,developed,and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and detection.The precision of the liquid transfer and temperature control was tested.A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process,from SARS-CoV-2 nucleic acid extraction to amplification,was evaluated. Results The precision of the syringe transfer volume was 19.2±1.9 μL(set value was 20),32.2±1.6(set value was 30),and 57.2±3.5(set value was 60).Temperature control in the amplification tube was measured at 60.0±0.0 ℃(set value was 60)and 95.1±0.2 ℃(set value was 95)respectively.SARS-Cov-2 nucleic acid extraction yield through the device was 7.10×106 copies/mL,while a commercial kit yielded 2.98×106 copies/mL.The mean time to complete the entire assay,from SARS-CoV-2 nucleic acid extraction to amplification detection,was 36 min and 45 s.The detection limit for SARS-CoV-2 nucleic acid was 250 copies/mL. Conclusion The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).
2.Establishment of rapid influenza virus detection technology based on an integrated system
Xiaoshan ZHANG ; Jing MA ; Yanzhe HAO ; Jing YANG ; Meiling HOU ; Hongxia LI ; Jingduan LIU ; Haodi MENG ; Yuxi CAO ; Xiaoguang ZHANG
Chinese Journal of Experimental and Clinical Virology 2024;38(3):319-325
Objective:This study aimed to develop a rapid and accurate integrated nucleic acid detection method tailored for the influenza virus.Methods:We designed primers and probes targeting the predominant influenza virus strains circulating in China in recent years. These were integrated with extraction and amplification reagents and a point of care testing (POCT) system to facilitate a seamless and expedited process involving nucleic acid extraction, reaction system preparation, amplification, and result interpretation for the influenza virus. The specificity of the POCT system was evaluated using cultured influenza viruses, while its cross-reactivity was assessed against common respiratory pathogens, including adenovirus and respiratory syncytial virus.Results:Our study successfully developed duplex amplification primers and probes for both influenza A and B viruses, achieving a detection threshold as low as 500 copies/ml. Specificity tests confirmed that the detection reagents did not show cross-reactivity with other respiratory pathogens such as adenovirus and respiratory syncytial virus. The POCT-based rapid nucleic acid detection method for influenza virus was established, it is capable of completing the entire process from nucleic acid extraction to amplification and result interpretation within 50 minutes, while enabling real-time data upload.Conclusions:The POCT-based rapid influenza virus detection kit developed in this study offers a " sample in, results out" convenience, making it suitable for rapid influenza virus detection in primary care settings. This innovation has significant potential for clinical application.
3.Progress of reversible parkinsonism
Ting PU ; Tao CHEN ; Mengyuan XU ; Xu MENG ; Jia TANG ; Xiaoguang LEI
Chinese Journal of Neurology 2022;55(7):775-782
Parkinsonism is a clinical syndrome caused by many reasons, mainly manifested as bradykinesia, stiffness, static tremor and postural instability. Common disease development patterns include occult onset, gradual development, and little natural remission. However, clinically there are some Parkinsonism that will improve, naturally alleviate or "cure", called reversible parkinsonism (RP). By searching the relevant literature, RP was classified into 12 different types: drugs induced, poisoning induced, infection induced, intracranial vascular induced, structural encephalopathy related, changes in intracranial pressure related, imbalance of internal environment induced, visceral diseases related, alcohol withdrawal related, surgery related, immunization and radiotherapy induced RP. This article aims to provide clinicians with more ideas for the clinical diagnosis and treatment of parkinsonism, so as to promote clinicians to make reasonable identification and diagnosis and treatment of parkinsonism as soon as possible.
4.Expert Consensus for Thermal Ablation of Pulmonary Subsolid Nodules (2021 Edition).
Xin YE ; Weijun FAN ; Zhongmin WANG ; Junjie WANG ; Hui WANG ; Jun WANG ; Chuntang WANG ; Lizhi NIU ; Yong FANG ; Shanzhi GU ; Hui TIAN ; Baodong LIU ; Lou ZHONG ; Yiping ZHUANG ; Jiachang CHI ; Xichao SUN ; Nuo YANG ; Zhigang WEI ; Xiao LI ; Xiaoguang LI ; Yuliang LI ; Chunhai LI ; Yan LI ; Xia YANG ; Wuwei YANG ; Po YANG ; Zhengqiang YANG ; Yueyong XIAO ; Xiaoming SONG ; Kaixian ZHANG ; Shilin CHEN ; Weisheng CHEN ; Zhengyu LIN ; Dianjie LIN ; Zhiqiang MENG ; Xiaojing ZHAO ; Kaiwen HU ; Chen LIU ; Cheng LIU ; Chundong GU ; Dong XU ; Yong HUANG ; Guanghui HUANG ; Zhongmin PENG ; Liang DONG ; Lei JIANG ; Yue HAN ; Qingshi ZENG ; Yong JIN ; Guangyan LEI ; Bo ZHAI ; Hailiang LI ; Jie PAN
Chinese Journal of Lung Cancer 2021;24(5):305-322
"The Expert Group on Tumor Ablation Therapy of Chinese Medical Doctor Association, The Tumor Ablation Committee of Chinese College of Interventionalists, The Society of Tumor Ablation Therapy of Chinese Anti-Cancer Association and The Ablation Expert Committee of the Chinese Society of Clinical Oncology" have organized multidisciplinary experts to formulate the consensus for thermal ablation of pulmonary subsolid nodules or ground-glass nodule (GGN). The expert consensus reviews current literatures and provides clinical practices for thermal ablation of GGN. The main contents include: (1) clinical evaluation of GGN, (2) procedures, indications, contraindications, outcomes evaluation and related complications of thermal ablation for GGN and (3) future development directions.
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5.Progress on quantitative assessments of motor symptoms for Parkinson′s disease
Mengyuan XU ; Tao CHEN ; Xu MENG ; Yuexian LIU ; Caifei YANG ; Xiaoguang LEI
Chinese Journal of Neurology 2020;53(10):845-854
Clinical scales and external devices are two important ways to quantify the severity of motor symptoms for Parkinson′s disease (PD). As one representative of scales, the Unified Parkinson′s Disease Rating Scale is considered the "gold standard" in the comprehensive assessment of PD. External devices are divided into electrophysiological and mechanical devices based on the type of sensors. Both sides can monitor the PD motor symptoms objectively. Currently, numerous and ununified parameters of devices make the clinical quantification of PD symptoms still rely on scales. This article reviewed the progress on quantitative assessments of motor symptoms for Parkinson′s disease.
6.Prognostic factors of moter function after surgery for patients with metastatic spinal cord compression: a multicenter retrospective cohort stduy
Yongheng LIU ; Xiaoguang YU ; Yongcheng HU ; Xionggang YANG ; Xuening MENG ; Dengxing LUN ; Feng WANG ; Mingyou XU ; Jiangtao FENG ; Kunchi HUA ; Li YANG ; Hao ZHANG ; Haoran ZHANG ; Zhaowan XU ; Dexiu SUN
Chinese Journal of Orthopaedics 2019;39(2):65-73
Objective To identify prognostic factors ofmotorfunctionafter surgery of metastatic spinal cord compression (MSCC).Methods The clinical data of 681 patients with spinal metastases from January 2008 to December 2017 were retrospectively analyzed.According to inclusion and exclusion criteria,a total of 206 patients with spinal metastatic were included.Postoperative neurological function was assessed using Frankel classification.The influence of age,gender,preoperative status,number of spine metastases,location of spinal metastases,visceral metastases,bone metastases,primary tumor type,interval from symptom to surgery,time of developing motor deficits,interval from primary tumor diagnosis to MSCC,preoperativethe Eastern Cooperative Oncology Group performance status (ECOG-PS),Karnofsky Performance score (KPS) and surgical procedures on postoperative function outcomes were explored.Results 140 (68.0%) patients were able to walk postoperatively compared with 88 (42.7%) patients preoperatively.Moreover,in 89.8% of all patients,79 ambulatory patients maintained ambulation after treatment.The univariate analysis according to Ordered-logit model showed thatnumber of spine metastases,location of spinal metastases,preoperative ECOG-PS,preoperative KPS,interval from symptom to surgery and time of developing motor deficits were related with posttreatment motor functions.The multivariable analysis showed that number of spine metastases (OR=2.03;95%CI:1.12-3.33;P=0.04),preoperative ECOG-PS (OR=4.84;95%CI:2.42-8.15;P=0.038),interval from symptom to surgery (OR=3.78;95%CI:3.12-9.15;P=0.024),time of developing motor deficits(OR=2.75;95%CI:1.22-3.89;P=0.01) were independent prognostic factors for function outcomes.Conclusion 1-2 levels of metastasis,Interval from symptom to treatment ≥ 48 h,time of developing motor deficits ≥7 d,and ECOG-PS 1-2 can be considered as the most significant positive prognosticfactors for post-treatment ambulatory status.Spinal metastasis should have a higher priority,and immediate intervention should be started before the development of irreversible neurologic deficits.Increasing awareness of early symptoms and earlier screeningwith regular outpatient review might make a difference for patients with MSCC.Consequently,the identified prognostic factors can be considered as apreoperative assessment tool to predict the neurologic outcomeand guide clinical treatment for individual patients with MSCC.
7. Characterization analysis of gM, gL genes of varicella zoster virus in six provinces of China
Junjie MOU ; Mengyuan QI ; Chongshan LI ; Huanying ZHENG ; Meng CHEN ; Jianhui ZHOU ; Jilan HE ; Wenbo XU ; Songtao XU ; Xiaoguang XU
Chinese Journal of Preventive Medicine 2018;52(4):419-423
Objective:
To analyze the genetic characterization of glycoprotein M(gM.),glycoprotein L(gL) of varicella zoster virus.
Methods:
According to the program of "Ministry of Science and Technology of China" , Based on the 12 suspected VZV patients monitored in Beijing (1 case), Shanghai (5 cases), Jilin (2 cases), Qinghai (1 case), Guangdong (2 case) and Sichuan (case) in 2007-2015. A total of 12 Vesicle fluid and throat swab samples were collected. Positive samples were identified by Agarose gel electrophoresis and two glycoprotein genes were amplified by polymerase chain reaction (PCR). Nucleotide sequences were determined and analyzed by PCR amplification of VZV positive specimens V-OKA-BK of the domestic varicella attenuated live vaccine and the Varilrix-1 of the imported attenuated live vaccine. Nucleotide sequences of VZV positive specimens, vaccine strains (V-OKA-BK, varilrix-1) and GenBank foreign wild strains (41 strains), parent strains (P-oka), vaccine strains (V-oka, Varilrix, Varivax) were compared using BioEdit and MEGA 5.0.
Results:
12 specimens were VZV positive. Compared with the vaccine strains and the parent strains, the GM gene of 1 positive specimen had radical mutation at 86686 sites, which resulted in amino acid mutation, 5 positive specimens had base mutation at 87844 sites, and 30 strains of foreign wild strains had the same variation at 87 844 sites. 1 positive specimens of gL gene in 101245 sites had base mutation, and led to amino acid mutation, 6 positive specimens at 101624, 101625, 101626 sites had base of loss and the foreign wild strains in these 3 sites had the same variation. Compared with the vaccine strains, the nucleotide and amino acid homology of gM of 12 VZV positive specimens were 99.2%-100% and 98.2%-100%, respectively, and gL of those were 99.3%-100% and 98.6%-100%, respectively. Compared with 41 strains of foreign wild strains, homology of gM's nucleotides and amino acid were 99.3%-100% and 98.5%-100%, respectively; 99.1%-100% and 98.6%-100% for gL. The results of phylogenetic analysis showed that 7 VZV positive samples were on the same branch with 4 vaccine strains and p-oka strain. Based on gL, 12 VZV positive samples were on the same branch as the vaccine strains and p-oka strain.
Conclusion
This study demonstrates that the genes of gM, gL are highly conserved and remain stable immunogen, which may be involved in the attenuation of VZV and need to be further researched.
8.The analysis of the changes of maternal thyroid autoantibodies during early pregnancy
Xiaoguang SHI ; Cheng HAN ; Chenyan LI ; Jinyuan MAO ; Weiwei WANG ; Xiaochen XIE ; Weiwei ZHOU ; Chenyang LI ; Lihua BI ; Tao MENG ; Shaowei ZHANG ; Jianling DU ; Zhengnan GAO ; Xiaomei ZHANG ; Chenling FAN ; Zhongyan SHAN ; Weiping TENG
Chinese Journal of Endocrinology and Metabolism 2016;32(5):376-379
Objective To investigate the changes and related factors of maternal thyroid autoantibodies during early pregnancy. Methods Urinary iodine concentration( UIC) , serum thyroid stimulating hormone( TSH) , free thyroxine ( FT4 ) , thyroid-peroxidase antibody ( TPOAb ) , thyroglobulin antibody ( TgAb ) concentrations were determined in 7 190 women during early pregnancy in an iodine-sufficient region of China. Results The prevalence of TPOAb positivity and TgAb positivity were 8. 7% and 12. 0% respectively. The prevalence of overt hypothyroidism and subclinical hypothyroidism increased significantly in group of thyroid antibody positivity. The prevalence of TPOAb positivity and TgAb positivity presented a U-shaped curve, ranging from mild iodine deficiency to iodine excess, especially increased significantly in the group with UIC<100 μg/L. Conclusion Prevalence of thyroid antibodies positivity became higher during early pregnancy. The positive thyroid autoantibodies during pregnancy were significantly associated with maternal hypothyroidism. Both iodine excess and iodine deficiency are risk factors of positive thyroid antibodies.
9.Effects of Buyanghuanwu decoction on the protein expression of PI3K, Akt, Bcl-2 and BAX in brain tissue of a rat model of cerebral hemorrhage
Xiaoguang WU ; Zhifu QIU ; Jie MENG ; Bingxue ZU ; Mengmeng LI ; Hui MIAO
Chinese Journal of Tissue Engineering Research 2016;20(40):5933-5938
BACKGROUND:Buyanghuanwu decoction has excel ent neuroprotective effect and can efficiently suppress nerve cel apoptosis caused by cerebral ischemia-reperfusion injury.
OBJECTIVE:To investigate the effect and mechanisms of Buyanghuanwu decoction on neuronal apoptosis around hematoma in cerebral hemorrhage rats.
METHODS:Seventy-two adult Sprague-Dawley rats were randomly divided into sham operation group, model group, Buyanghuanwu decoction group, and Ginkgo biloba group. Except the sham operation group, rat models of cerebral hemorrhage were established in other three groups. At 2 days after modeling, rats in the Buyanghuanwu group and Ginkgo biloba group were given Buyanghuanwu decoction 26 g/(kg?d)and Ginkgo biloba 3.5 mg/(kg?d) daily by gavage, for 14 consecutive days. Rats in the sham operation group and model group received an equal volume of saline for 14 consecutive days. After the last administration, brain tissue was obtained. TUNEL assay was utilized to detect neuronal apoptosis. Immunohistochemistry was used to detect PI3K, Akt, Bcl-2, and BAX protein expression. Wet and dry weight method was used to detect brain water content. Evans Blue assay was utilized to determine blood-brain barrier permeability.
RESULTS AND CONCLUSION:(1) Compared with the sham operation group, the number of apoptotic neurons, brain water content, Evans blue content and PI3K, Akt, Bcl-2, BAX protein expression increased in the model group (P<0.05). (2) Compared with the model group, the number of apoptotic neurons, BAX protein expression, brain water content and Evans blue content were significantly reduced in the Buyanghuanwu group and Ginkgo biloba group (P<0.05), but PI3K, Akt and Bcl-2 protein expression was significantly increased (P<0.05). (3) Results suggested that Buyanghuanwu decoction inhibited neuronal apoptosis and protected brain tissue by reducing blood-brain barrier permeability, cerebral edema, and by activating PI3K/Akt signaling pathway, regulating Bcl-2 and BAX protein expression ratio.
10.Fast Structure Confirmation of Daptomycin by Matrix_assisted Laser Desorption Ionization Mass Spectrometry and Electrospray Ionization Mass Spectrometry
Yajuan HUANG ; Tuo ZHANG ; Zhiguo HAN ; Xiaoguang MENG ; Chunyan SONG ; Shuchen LIU ; Junjie ZHENG ; Kaihua WEI
Chinese Journal of Analytical Chemistry 2015;(1):63-68
Matrix_assisted laser desorption ionization_time of flight tandem mass spectrometry ( MALDI_TOF/TOF MS) and electrospray ionization_quadrupole_time of flight mass spectrometry ( ESI_Q_TOF MS) were used to confirm the structure of cyclic lipopeptide daptomycin fastly. First, the relative molecular weight 1916. 7107 of daptomycin was measured by ESI with error 0. 0007. The sample’s doubly charged peak m/z 809. 848 was selected as precursor ion for ESI_MS/MS analysis, and the exocyclic amino acid sequence C9 H19 CO_Trp_Asn_Asp was successfully matched. Second, the experimental conditions of cleaving daptomycin by lithium hydroxide ( LiOH) were optimized and the ring_opened process was monitored by MALDI_TOF/TOF MS. After obtaining ring_opened product with purity of above 95%, the MS/MS measurements by MALDI and ESI were carried out. The b+and y+of ring_opened product were completely matched, which confirmed the amino acid sequence of daptomycin. Finally, ESI_MS/MS conditions of ring_opened product were further optimized to obtain more low mass fragment ions for analyzing the structure of fatty acid chain and the cleavage pattern of fat chain in mass spectrometry was proposed. The method was fast, convenient, accurate and reliable for identifying cyclic lipopeptide compounds.

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