1.Evaluation of sensitivity of SARS-CoV-2 antigen-detecting rapid diagnostic cards to different strains
Chinese Journal of Biologicals 2023;36(6):680-686
Objective To compare the sensitivity(dilution) of antigen-detecting rapid diagnostic cards for severe acute respiratory symptom coronavirus 2(SARS-CoV-2) at home and abroad to different strains.Methods Vaccine bulks of four SARS-CoV-2 strains(original strain,Beta,Delta and Omicron) produced by Wuhan Institute of Biological Products Co.,Ltd.were used as the sample panel for sensitivity assessment,of which a series of diluted samples were detected by using 21 batches of SARS-CoV-2 antigen-detecting rapid diagnostic cards from 17 domestic and foreign manufacturers and SARS-CoV-2 nucleic acid detection reagent from Shanghai GeneoDx Biotech Co.,Ltd,respectively.The sensitivity of antigendetecting rapid diagnostic cards and nucleic acid detection reagent was evaluated according to the dilutions.The results of SARS-CoV-2 antigen-detecting rapid diagnostic reagents and nucleic acid detection reagent were compared to determine the nucleic acid detection Ct value corresponding to the group of antigen-detecting rapid diagnostic reagent with the highest dilution,namely the highest sensitivity.Results The sensitivity of antigen detection cards for the vaccine bulks of original strain,Beta,Delta and Omicron was 1:10~1:8 × 10~4.1:10~3~1:2 × 10~5,1:10~2~1:4 × 10~4,and 1:10~1:4 × 10~5,respectively;The sensitivity of nucleic acid detection cards was 10~(-6),10~(-5),10~(-4) and 10~(-7),respectively.The Ct values of N gene which were reached by high sensitivity antigen-detecting rapid diagnostic cards were as follows:original strain(10~(-4)) of more than 31,Beta variant(10~(-5)) of more than 36,Delta variant(10~(-4)) of more than 34,Omicron variant(10~(-5)) of more than 33,meeting the requirements of domestic and European Union for SARS-CoV-2 antigen-detecting rapid diagnostic cards.Conclusion All the antigen-detecting rapid diagnostic cards detected the four virus strains,while the sensitivity of different reagents to different variants varies to some extent,among which the sensitivity to Omicron variant varies the most.
2.Let talk teaching into pathophysiologic teaching
Fang XU ; Ying LIU ; Lei SHI ; Hongjing CAI ; Weichen WANG ; Jinrong CHEN
Chinese Journal of Medical Education Research 2005;0(05):-
Talk teaching is an effective form of teaching research activity.It focuses on the teachers,teaching ideas and their abilities of curriculum development,curriculum designing and teaching practice.It is of great significance in optimizing pathophysiologic teaching,improving the teaching effect and teachers'quality.It is worth discussing and promoting.Talk teaching should be the basic skill necessary to the teachers of basic medicine.
3.The application effect of enhanced recovery after surgery in laparoscopic colorectal cancer surgery in primary hospitals
Chunliang WANG ; Ruihua QI ; Ying XU ; Xiao WANG ; Yunfeng ZHU ; Jinrong ZHAO ; Qing SHI
Chinese Journal of Geriatrics 2021;40(4):483-486
Objective:To evaluate the application efficacy of enhanced recovery after surgery(ERAS)in laparoscopic colorectal cancer surgery in primary hospitals.Methods:A total of 116 patients who underwent laparoscopic colorectal cancer surgery from January 2017 to December 2018 at our hospital were enrolled in this study.According to the perioperative rehabilitation program, 116 patients were divided into the group A(n=67, receiving enhanced recovery after surgery)and the group B(n=49, receiving traditional recovery after surgery).Results:The incidences of preoperative thirst and hunger were lower in the group A than in the group B(11.9% vs.53.1%, 16.4% vs.51.0%, χ2=23.10 and 15.83, respectively, P<0.001). The levels of CRP and blood glucose in the two groups were significantly higher after operation than before operation, and reached the peak values on the 3rd day after the operation.At different time points after operation, CRP levels and blood glucose levels were higher in the group B than in the group A(all P<0.05). On the 7th day after operation, blood glucose level was recovered to the preoperative level in the group A, while it was not so in the group B. The incidence of complication in the group A was similar to the group B(7.46% vs.12.2%, χ2=0.75, P>0.05). The hospitalization period was shorter and the hospitalization cost was less in the group A than in the group B(8.16±1.33)d vs.(15.39±2.81)d, (46100±1800)yuan vs.(56900±5600)yuan, t=10.98 and 9.96, P=0.000). Conclusions:The application of enhanced recovery after surgery is beneficial for perioperative safety, can reduce surgical stress response, promote postoperative recovery, shorten hospitalization time after surgery and reduce hospitalization costs in laparoscopic colorectal cancer surgery.
4.Multidrug Resistance Mechanisms of Carbapenem Resistant Klebsiella pneumoniae Strains Isolated in Chongqing, China.
Jinrong YAN ; Shuli PU ; Xiaojiong JIA ; Xiuyu XU ; Shuangshuang YANG ; Jing SHI ; Shan SUN ; Liping ZHANG
Annals of Laboratory Medicine 2017;37(5):398-407
BACKGROUND: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is considered a serious global threat. However, little is known regarding the multidrug resistance (MDR) mechanisms of CRKP. This study investigated the phenotypes and MDR mechanisms of CRKP and identified their clonal characteristics. METHODS: PCR and sequencing were utilized to identify antibiotic resistance determinants. Integron gene cassette arrays were determined by restriction fragment length polymorphism (RFLP) analysis. Multi-locus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used for epidemiological analysis. Plasmids were typed by using a PCR-based replicon typing and analyzed by conjugation and transformation assays. RESULTS: Seventy-eight strains were identified as resistant to at least one carbapenem; these CRKP strains had a high prevalence rate (38.5%, 30/78) of carbapenemase producers. Additionally, most isolates harbored MDR genes, including Extended spectrum β-lactamases (ESBLs), AmpC, and quinolone and aminoglycoside resistance genes. Loss of porin genes was observed, and Class 1 integron was detected in 66.7% of the investigated isolates. PFGE and MLST results excluded the occurrence of clonal dissemination among these isolates. CONCLUSIONS: A high prevalence of NDM-1 genes encoding carbapenem resistance determinants was demonstrated among the K. pneumoniae isolates. Importantly, this is the first report of bla(NDM-1) carriage in a K. pneumoniae ST1383 clone in China and of a MDR CRKP isolate co-harboring bla(NDM-1), bla(KPC-2), bla(CTX-M), bla(SHV), acc(6′)-Ib, rmtB, qnrB, and acc(6′)-Ib-cr.
China*
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Clone Cells
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Drug Resistance, Bacterial
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Drug Resistance, Microbial
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Drug Resistance, Multiple*
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Electrophoresis, Gel, Pulsed-Field
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Genes, MDR
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Integrons
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Klebsiella pneumoniae*
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Klebsiella*
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Molecular Epidemiology
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Phenotype
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Plasmids
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Pneumonia
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Polymerase Chain Reaction
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Polymorphism, Restriction Fragment Length
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Prevalence
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Replicon
5. Solitary fibrous tumor/hemangiopericytoma of central nervous system: a clinicopathological analysis of 60 cases
Yu GUAN ; Xuan WANG ; Nan WU ; Zhenfeng LU ; Qunli SHI ; Jinrong WU
Chinese Journal of Pathology 2019;48(1):31-36
Objective:
To investigate the clinical manifestations, imaging features, clinicopathologic features, and differential diagnosis of solitary fibrous tumors/anginoblastomas (SFT/HPCs) originating in the central nervous system.
Methods:
Sixty cases of SFT/HPCs originating in the central nervous system were collected at Nanjing Jinling Hospital, from January 1, 2008 to December 31, 2016. The clinical data, imaging data, histomorphologic changes and immunohistochemical finding were analyzed in the sixty cases.
Results:
The 60 cases included 26 males and 34 females, aged 14 to 85 (median 49) years. The main clinical manifestations were headache, dizziness with nausea and vomiting. Radiologically, the tumors were large, enhancing, solid and cystic masses attached to the dura. Histopathologically, the neoplasms were composed of spindle cells with oval nuclei, inconspicuous nucleoli and moderate amount of eosinophilic cytoplasm arranged in fascicles with areas of hyalinized stroma, myxoid changes and a staghorn vascular pattern. Immunohistochemically, tumor cells of all cases were positive for vimentin (100.0%, 60/60), STAT6 (98.3%, 59/60), CD34 (61.7%, 37/60), and the tumor cells were typically positive for CD99, bcl-2, EMA and SSTR2 as well.Negative for S-100 protein, SOX10, E-cadherin, GFAP. Ki-67 index ranged from 1% to 50%. Forty cases were followed up for 6 to 82 months with average of 40 months, 30 patients were alive and 10 patients died.
Conclusions
Central nervous system SFT/HPCs can be aggressive and relapses may occur several years after diagnosis. STAT6 is highly sensitive and specific for the diagnosis. Complete tumor resection is optional treatment followed by radiotherapy and chemotherapy. There is a correlation between the prognosis and the location of the disease, the histological grade, Ki-67 index, and fusion gene variants.
6.Cloning and analyzing of MDCK cells for H5N1 influenza vaccine production
Xiaoyuan HUANG ; Jiayou ZHANG ; Yang LIU ; Wei ZHAO ; Jinrong SHI ; Xinru LE ; Gang YU ; Xixin HAN ; Xiaoming YANG
Chinese Journal of Microbiology and Immunology 2019;39(2):81-87
Objective To screen a Madin-Darby canine kidney (MDCK) cell line for H5N1 influ-enza virus isolation and to evaluate its safety in vaccine production. Methods MDCK cells were cloned by the method of limiting dilution. Hemagglutination test was used to screen MDCK cells that were suitable for H5N1 influenza virus production. Tests for analyzing the characteristics, extraneous agents, endogenous agents and tumorigenicity of MDCK cells were performed according to Chinese Pharmacopeia Volume Ⅲ. Results A total of 108 MDCK cell lines were obtained and three of them were selected after hemagglutina-tion test. G1 cells were chosen following further screening with tumorigenicity test and receptor abundance analysis. The average number of chromosomes of the MDCK-G1 cells was 78±4. No bacteria, fungi or myco-plasma contamination was detected. In experimental group, each nude mouse was injected with 1×107/ml viable cells to observe their tumorigenicity. Twelve weeks after cell injection, no node was found at injection sites or in gross anatomy. There was no significant difference between the experimental and negative control groups. The result of the tumorigenicity test was negative. No node formation was found after injecting nude mice with cell lysate or cellular DNA collected from equivalent amount of cells. It was indicated that the MDCK-G1 cells were of low-oncogenic potential. Conclusions The MDCK-G1 cell line could be used as a substrate to produce H5N1 influenza virus vaccine.
7.A multi-center study on the normal range of exhaled nitric oxide in 6-18-year-old children in China
Hao ZHANG ; Wenhui JIANG ; Chunyan MA ; Yongsheng SHI ; Chunmei JIA ; Jinrong WANG ; Yuling HAN ; Yuehua ZHANG ; Ming LI ; Fei WANG ; Yanyan YU ; Yufen WU ; Yong FENG ; Li LIU ; Aihong LIU ; Qiaoling ZHANG ; Zhen LONG ; Fuli DAI ; Yanli ZHANG ; Minghong JI ; Dongjun MA
Chinese Journal of Applied Clinical Pediatrics 2020;35(21):1618-1623
Objective:To investigate the normal range of exhaled nitric oxide (FeNO) in 6-18-year-old children in China, so as to provide a data base for the establishment of FeNO standards for Chinese children.Methods:A multi-center study was conducted on 5 949 children aged 6-18 (3 101 males and 2 848 females) in 16 pro-vinces of 7 administrative districts in China.According to the technical standard recommended by American Thoracic Society/European Respiratory Association, FeNO was measured, and the relationship of FeNO with the sex, age, height, weight, body mass index and region was discussed.Results:The geometric mean FeNO value of Chinese children aged 6-18 was 14.1 ppb, and its 95% confidence interval (skewness distribution) was 1.0-38.2 ppb.The geometric mean FeNO values of children aged 6-11 and 12-18 were 13.1 ppb and 15.7 ppb, respectively, and their 95% confidence intervals (skewness distribution) were 1.0-38.1 ppb and 2.0-38.2 ppb.For children at and under 11 years old, FeNO decreased with age, with a mean decline of 1 ppb per year.The multiple linear regression results suggested that there was a significant correlation between FeNO and age for children aged 6-11, and FeNO of children aged 12-18 was significantly correlated with the gender, height, and region(all P<0.01). Conclusions:FeNO values of Chinese children and adolescents in this study are higher than those obtained by the previous study conducted from 2010 to 2012.For children aged 12-18, 16 ppb is recommended as the clinical cut-off point.For children at or under 11 years old, the influence of age on FeNO should be considered, and the cut-off point of FeNO decreases by 1 ppb as the age is reduced by one year.
8.Mechanism of Cuscutae Semen to Control Premature Ovarian Failure Based on Lipid and Atherosclerosis Pathways
Ying XIE ; Min XIAO ; Xiaocui JIANG ; Jinrong ZHANG ; Ruoyi SHI ; Jiawei ZHANG ; Min ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):109-118
ObjectiveThe active ingredients, action targets, and signaling pathways of Cuscutae Semen to control premature ovarian failure were initially predicted by network pharmacology and molecular docking techniques, and an animal model of premature ovarian failure was constructed to explore the mechanism of Cuscutae Semen based on lipid and atherosclerosis signaling pathways. MethodThe effective components and corresponding targets of drugs were obtained from Traditional Chinese Medicines Systems Pharmacology Platform (TCMSP), Swiss Target Prediction, Pharmmapper, and other databases. GeneCards database was used to collect disease-related targets. Venny2.1.0 online tool was used to screen out the intersection targets of drugs and diseases, and STRING database and Cytoscape v3.7.2 software were used to construct the network diagram of "drug-component-target" and protein-protein interaction (PPI). The gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses of the intersection targets were performed by running the R language script. The molecular docking technology was utilized to dock drug components with targets and visualize some of the docking results. The mice were randomly divided into a blank group, a model group, a Cuscutae Semen group, and an estradiol valerate group, and the ovarian premature failure model was prepared by chronic stress. The blank group and the model group were gavaged with the same amount of normal saline, and the Cuscutae Semen group was given a Cuscutae Semen decoction of 2.6 g·kg-1·d-1. The estradiol valerate group was given an estradiol valerate solution of 0.13 mg·kg-1·d-1. After four weeks, samples were collected, and hematoxylin-eosin (HE) staining was performed to observe the histopathological changes in the ovary. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), Muller's tube inhibitor/anti-Muller's tube hormone (AMH), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were determined by enzyme-linked immunosorbent assay (ELISA). The expression levels of extracellular regulatory protein kinase (ERK), nuclear transcription factor-κB p65 (NF-κB p65), nuclear transcription factor-κB suppressor α (IκBα), interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) were measured by Western blot. ResultA total of 171 targets of Cuscutae Semen for the prevention and treatment of premature ovarian failure were screened, mainly including tumor protein p53 (TP53), protein kinase B1 (Akt1), sarcoma (SRC), tumor necrosis factor (TNF), epidermal growth factor receptor (EGFR), etc. KEGG pathway enrichment analysis predicts that Cuscutae Semen is mainly involved in lipid and atherosclerosis, TNF signaling pathway, and TP53 signaling pathway to control premature ovarian failure. The animal experiments show that compared with the premature ovarian failure model group, the Cuscutae Semen group can significantly upregulate AMH, E2, and HDL-C (P<0.05, P<0.01), significantly downregulate LH, TC, and LDL-C (P<0.01), greatly reduce IL-1β, IL-6, and TNF-α protein levels, as well as ERK, NF-κB p65, and their phosphorylation levels (P<0.01). ConclusionCuscutae Semen can regulate hormone levels and improve ovarian function through a multi-component, multi-target, and multi-pathway approach, and the mechanism may be related to the regulation of lipid and atherosclerosis signaling pathways.
9.Effect of Glycyrrhizae Radix et Rhizoma-containing Serum on LPS-induced Inflammation in Caco2 Cells Based on Inhibition of Ferroptosis by Nrf2/HO-1 Pathway
Jinrong KONG ; Gaoxiang SHI ; Jing HOU ; Ye FENG ; Qingzhen XIANG ; Yunlai WANG ; Zihua XUAN ; Fan XU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):144-153
ObjectiveTo investigate the effect of Glycyrrhizae Radix et Rhizoma (GR)-containing serum on lipopolysaccharide (LPS)-induced inflammation in human colon epithelial adenocarcinoma cells (Caco2) based on inhibition of ferroptosis by the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. MethodCaco2 cells were divided into a normal group, a model group (LPS, 200 μg·L-1), low-, medium-, and high-dose GR-containing serum groups (5%, 10%, 20%), and a ferroptosis inhibitor group (3-amino-4-cyclohexylamino-benzoic acid ethyl ester, Fer-1, 10 μmol·L-1). The cells in the normal group were cultured normally, while those in other groups underwent the induction of an inflammation model. The cells in the low-, medium-, and high-dose GR-containing serum groups were treated with 5%, 10%, and 20% GR-containing serum for 24 hours, respectively, and the cells in the ferroptosis inhibitor group were treated with Fer-1 for 24 hours. Transmission electron microscopy was used to observe mitochondrial morphology in each group. Flow cytometry was used to detect intracellular Fe2+ levels. Microplate assays were performed to measure superoxide dismutase (SOD) activity, malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) levels. Enzyme-linked immunosorbent assay (ELISA) was used to measure interleukin-1β (IL-1β), IL-6, IL-10, and tumor necrosis factor-α (TNF-α) levels. Western blot was used to measure the expression levels of Nrf2, HO-1, ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GSH-Px4) proteins. Small interfering RNA (siRNA) was used to investigate the role of Nrf2 in ferroptosis regulation. The cells after interference were divided into a negative control (NC) group, a Si-Nrf2 group, a GR-containing serum (20%) + Si-Nrf2 group, and a GR-containing serum (20%) + NC group. Microplate assays were performed to measure MDA, SOD, and GSH-Px levels, and Western blot was used to measure the expression levels of Nrf2, HO-1, FTH1, and GSH-Px4 proteins. ResultCompared with the normal group, the model group showed mitochondrial contraction, increased mitochondrial membrane thickness, and smaller mitochondrial morphology, increased Fe2+ content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px expression (P<0.01), increased MDA content (P<0.01), reduced expression levels of Nrf2 and HO-1 (P<0.05), reduced FTH1 expression (P<0.01), and down-regulated GSH-Px4 expression (P<0.01). In the GR-containing serum groups, the medium- and high-dose groups showed a significant decrease in Fe2+ content (P<0.01), potentiated SOD and GSH-Px activities (P<0.01), and decreased MDA levels (P<0.01). The high-dose group showed a significant increase in Nrf2 expression (P<0.05), and the medium-dose group showed increased expression of HO-1 and GSH-Px4 proteins (P<0.05). The expression levels of FTH1 significantly increased in the low-, medium-, and high-dose groups (P<0.01). The study on mechanism revealed that compared with the NC group, the cells transfected with Nrf2 siRNA showed increased MDA content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px activity (P<0.01), decreased expression of Nrf2 and HO-1 (P<0.01), and reduced levels of FTH1 and GSH-Px4 proteins (P<0.01). Compared with the Si-Nrf2 group, the cells treated with GR-containing serum showed a decrease in MDA content (P<0.01), an increase in SOD activity (P<0.01), an increase in GSH-Px activity (P<0.01), increased expression of Nrf2 and FTH1 proteins (P<0.05), and higher expression levels of HO-1 and GSH-Px4 proteins (P<0.01). ConclusionGR-containing serum can reduce the inflammatory cytokines and oxidative stress levels in LPS-induced Caco2 cells. Its mechanism is related to the promotion of Nrf2/HO-1 signaling pathway expression, alleviating intracellular lipid peroxidation and inhibiting ferroptosis.
10.Gold nanorods together with HSP inhibitor-VER-155008 micelles for colon cancer mild-temperature photothermal therapy.
Xichuan TANG ; Liwei TAN ; Kun SHI ; Jinrong PENG ; Yao XIAO ; Wenting LI ; Lijuan CHEN ; Qian YANG ; Zhiyong QIAN
Acta Pharmaceutica Sinica B 2018;8(4):587-601
Enhancing the heat-sensitivity of tumor cells provides an alternative solution to maintaining the therapeutic outcome of photothermal therapy (PTT). In this study, we constructed a therapeutic system, which was composed of methoxy-polyethylene-glycol-coated-gold-nanorods (MPEG-AuNR) and VER-155008-micelles, to evaluate the effect of VER-155008 on the sensitivity of tumor cells to heat, and further investigate the therapeutic outcome of MPEG-AuNR mediated PTT combined with VER-155008- micelles. VER-155008- micelles down-regulate the expression of heat shock proteins and attenuate the heat-resistance of tumor cell. The survival of HCT116 cells treated with VER-155008- micelles under 45 °C is equal to that treated with high temperature hyperthermia (55 °C) . Furthermore, we proved either the MPEG-AuNR or VER-155008- micelles can be accumulate in the tumor site by photoacoustic imaging and fluorescent imaging. anti-cancer evaluation showed that tumor size remarkably decreased (smaller than 100 mm or vanished) when treated with combing 45 °C mild PTT system, which contrasted to the tumor size when treated with individual 45 °C mild PTT (around 500 nm) or normal saline as control (larger than 2000 nm). These results proved that the VER-155008- micelles can attenuate the heat-resistance of tumor cells and enhance the therapeutic outcome of mild-temperature photothermal therapy.