1.Wound Repairing Mechanomedicine
Zhixing LAN ; Yuchen WANG ; Zhihao GAO ; Huicong DU ; Yuyao LIN ; Maoguo SHU ; Jing LI ; Tianjian LU ; Feng XU ; Hao LIU
Journal of Medical Biomechanics 2025;40(3):749-759
The application of mechanics in clinical wound healing has a long history;however,the systematic underlying mechanisms remain unclear.With recent advancements in biomechanics and mechanobiology,the principles regarding how mechanical factors influence the formation,progression,and healing of wounds have gradually been elucidated.Herein,based on progress in theories,technologies,and clinical practices concerning the interplay between mechanics and wound healing,this study introduces the concept of wound-repairing mechanomedicine.Relevant research is systematically reviewed from the perspectives of biomechanics,mechanobiology,and mechanotherapy.Additionally,potential future development directions are prospectively analyzed to provide novel insights into wound care and strategies for preventing scar formation.
2.Influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts
Menglu WU ; Rui WANG ; Xinnan ZHENG ; Juan WU ; Lin HE ; Jiansheng DIAO ; Maoguo SHU ; Huicong DU
Chinese Journal of Burns 2025;41(4):355-363
Objective:To investigate the influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts (Fbs).Methods:This study was an experimental research. The keloid tissue was collected from 7 keloid patients (2 males and 5 females, aged 20-65 years, with a disease course of more than 1 year) who underwent keloid excision surgery at the Department of Plastic, Cosmetic and Maxillofacial Surgery of the First Affiliated Hospital of Xi'an Jiaotong University from September 2020 to September 2023. The primary Fbs were isolated and cultured, and cells from passages 3 to 6 were used for experiments. The cells were divided into control group and metformin group, and were cultured in complete medium. The medium for metformin group was supplemented with metformin at a final molarity of 60 mmol/L. The cell counting kit-8 was used to assess the proliferation activity of cells in two groups after 12 and 24 hours of culture, and the proliferation inhibition rate of cells in metformin group after 12 and 24 hours of culture was calculated, with a sample size of 6. The apoptosis detection kit was used to detect the apoptotic distribution of cells in control group after 0 hour (immediately) of culture and in metformin group after 12 and 24 hours of culture, with a sample size of 3. The cell cycle detection kit was used to detect the cycle distribution of cells in two groups after 12 and 24 hours of culture, with a sample size of 3. The eukaryotic mRNA sequencing was performed on suitable number of cells of two groups after 24 hours of culture, and the Kyoto encyclopedia of genes and genomes functional annotation analysis and functional enrichment analysis were performed after screening for differentially expressed genes (DEGs) with significantly differential expression between two groups. Western blotting was conducted to detect the protein expressions of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), and phosphorylated mammalian target of rapamycin (p-mTOR) in the PI3K/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway of cells in two groups after 24 hours of culture, with a sample size of 3.Results:After 12 and 24 hours of culture, the proliferation activity of cells in metformin group was significantly lower than that in control group (with t values of 4.70 and 24.02, respectively, P<0.05); the proliferation activity of cells in metformin group after 24 hours of culture was significantly lower than that after 12 hours of culture within the group ( t=4.73, P<0.05). Compared with that after 12 hours of culture within the group, the proliferation inhibition rate of cells in metformin group was significantly increased after 24 hours of culture ( t=5.29, P<0.05). Compared with that in control group after 0 hour of culture, the proportion of early apoptotic cells in metformin group was significantly increased (with t values of 6.62 and 4.58, respectively, P<0.05), and the proportion of early and late apoptotic cells was significantly increased after 12 and 24 hours of culture (with t values of 4.84 and 3.75, respectively, P<0.05). After 24 hours of culture, the proportion of late apoptotic cells in metformin group was significantly higher than that after 12 hours of culture within the group ( t=4.55, P<0.05). After 12 hours of culture, the proportion of S-phase cells in metformin group was significantly lower than that in control group ( t=5.90, P<0.05). After 24 hours of culture, compared with that in control group, the proportion of G0/G1-phase cells in metformin group was significantly increased ( t=5.36, P<0.05), while the proportion of G2/M-phase cells was significantly decreased ( t=17.63, P<0.05). The proportion of S-phase cells in metformin group after 24 hours of culture was significantly higher than that after 12 hours of culture within the group ( t=7.60, P<0.05). After 24 hours of culture, 4 814 DEGs with significantly differential expression were detected in the cells of metformin group compared with control group. The significantly upregulated and downregulated DEGs were mainly involved in biological functions related to signal transduction, cell growth and death, transport and catabolism, the endocrine system, the immune system, and cancer. The pathways that were significantly enriched with DEGs with significantly differential expression included the cell cycle and DNA replication, with the highest number of genes in the PI3K/Akt signaling pathway. After 24 hours of culture, the protein expressions of PI3K, p-Akt, and p-mTOR of cells in metformin group were 0.190±0.017, 0.170±0.017, and 0.247±0.005, respectively, which were significantly lower than 0.440±0.026, 0.300±0.060, and 0.547±0.025 in control group (with t values of 13.69, 3.61, and 20.12, respectively, P values all <0.05). Conclusions:Metformin can significantly inhibit the proliferation of human keloid Fbs through the PI3K/Akt/mTOR signaling pathway and effectively induce its apoptotic process, thereby exerting antifibrotic effects.
3.Construction and validation of predictive model for acute respiratory failure in adult patients with community-acquired pneu-monia
Ziming WANG ; Yue QU ; Dandan LI ; Huicong ZHOU ; Binbin WU ; Wei YU
Chinese Journal of Clinical Laboratory Science 2025;43(8):586-590
Objective To explore the risk factors of acute respiratory failure(ARF)in adult patients with community-acquired pneu-monia(CAP),and thereby construct and validate the efficacy of nomogram model.Methods The clinical and laboratory data of 172 adult CAP patients admitted to Taikang Xianlin Drum Tower Hospital affiliated to Nanjing University School of Medicine from January 2018 to December 2021 were retrospectively collected.The patients were divided into two groups based on whether they had concurrent ARF.After the comparison for the differences of single factor between the two groups,collinearity analysis was assessed.The risk fac-tors were then screened by binary logistic regression analysis with forward stepwise regression method.A nomogram model was subse-quently constructed and the discrimination and accuracy of the model were evaluated by ROC and colibration curves.Results Among the 172 CAP patients,53 cases(30.8%)developed ARF.The results of univariate analysis showed that the CAP patients with concur-rent ARF group had higher age,CURB-65 score and inflammatory markers than the non-concurrent ARF group,and the incidence of complex infection(culturing two or more pathogenic bacteria)was high.The values of CRP(C-reactive protein)and BUN/Alb(blood urea nitrogen/albumin)were significantly different between the two groups(53.910[25.900,101.200]vs.23.300[6.800,48.930],0.231[0.160,0.302]vs.0.123[0.089,0.171],P<0.05).Multivariate analysis indicated:glucose(Glu)≥6.06 mmol/L(odds ra-tio(OR):2.737,95%confidence interval(CI):1.116-7.037),AST(aspartate aminotransferase)≥22.5 U/L(OR:4.291,95%CI:1.779-11.120),fibrinogen(Fib)≤3.83 g/L(OR:3.955,95%CI:1.631-10.237),uric acid(UA)188.07 μmol/L(OR:4.617,95%CI:1.859-12.489),BUN/Alb≥0.15 mmol/g(OR:6.381,95%CI:2.423-18.513),total number of multicomor-bidity≥3(OR:6.191,95%CI:2.088-21.905)were the risk factors(P<0.05).All the screened indicators were incorporate into the nomogram model and its efficacy was verified.The results showed that the area under the curve of the model was 0.888[95%CI:0.840-0.935](P<0.05),the sensitivity was 0.868,and the specificity was 0.790.The calibration curve showed that the predicted probability of adult CAP patients-associated with ARF was in good consistency with the observed probability(Briser Score:0.125;H-L test:x2=7.563,P=0.477).Conclusion The established model has a good ability to predict adult CAP associated with ARF,and can provide a reference basis for early clinical prediction and intervention treatment.
4.Construction and validation of predictive model for acute respiratory failure in adult patients with community-acquired pneu-monia
Ziming WANG ; Yue QU ; Dandan LI ; Huicong ZHOU ; Binbin WU ; Wei YU
Chinese Journal of Clinical Laboratory Science 2025;43(8):586-590
Objective To explore the risk factors of acute respiratory failure(ARF)in adult patients with community-acquired pneu-monia(CAP),and thereby construct and validate the efficacy of nomogram model.Methods The clinical and laboratory data of 172 adult CAP patients admitted to Taikang Xianlin Drum Tower Hospital affiliated to Nanjing University School of Medicine from January 2018 to December 2021 were retrospectively collected.The patients were divided into two groups based on whether they had concurrent ARF.After the comparison for the differences of single factor between the two groups,collinearity analysis was assessed.The risk fac-tors were then screened by binary logistic regression analysis with forward stepwise regression method.A nomogram model was subse-quently constructed and the discrimination and accuracy of the model were evaluated by ROC and colibration curves.Results Among the 172 CAP patients,53 cases(30.8%)developed ARF.The results of univariate analysis showed that the CAP patients with concur-rent ARF group had higher age,CURB-65 score and inflammatory markers than the non-concurrent ARF group,and the incidence of complex infection(culturing two or more pathogenic bacteria)was high.The values of CRP(C-reactive protein)and BUN/Alb(blood urea nitrogen/albumin)were significantly different between the two groups(53.910[25.900,101.200]vs.23.300[6.800,48.930],0.231[0.160,0.302]vs.0.123[0.089,0.171],P<0.05).Multivariate analysis indicated:glucose(Glu)≥6.06 mmol/L(odds ra-tio(OR):2.737,95%confidence interval(CI):1.116-7.037),AST(aspartate aminotransferase)≥22.5 U/L(OR:4.291,95%CI:1.779-11.120),fibrinogen(Fib)≤3.83 g/L(OR:3.955,95%CI:1.631-10.237),uric acid(UA)188.07 μmol/L(OR:4.617,95%CI:1.859-12.489),BUN/Alb≥0.15 mmol/g(OR:6.381,95%CI:2.423-18.513),total number of multicomor-bidity≥3(OR:6.191,95%CI:2.088-21.905)were the risk factors(P<0.05).All the screened indicators were incorporate into the nomogram model and its efficacy was verified.The results showed that the area under the curve of the model was 0.888[95%CI:0.840-0.935](P<0.05),the sensitivity was 0.868,and the specificity was 0.790.The calibration curve showed that the predicted probability of adult CAP patients-associated with ARF was in good consistency with the observed probability(Briser Score:0.125;H-L test:x2=7.563,P=0.477).Conclusion The established model has a good ability to predict adult CAP associated with ARF,and can provide a reference basis for early clinical prediction and intervention treatment.
5.Wound Repairing Mechanomedicine
Zhixing LAN ; Yuchen WANG ; Zhihao GAO ; Huicong DU ; Yuyao LIN ; Maoguo SHU ; Jing LI ; Tianjian LU ; Feng XU ; Hao LIU
Journal of Medical Biomechanics 2025;40(3):749-759
The application of mechanics in clinical wound healing has a long history;however,the systematic underlying mechanisms remain unclear.With recent advancements in biomechanics and mechanobiology,the principles regarding how mechanical factors influence the formation,progression,and healing of wounds have gradually been elucidated.Herein,based on progress in theories,technologies,and clinical practices concerning the interplay between mechanics and wound healing,this study introduces the concept of wound-repairing mechanomedicine.Relevant research is systematically reviewed from the perspectives of biomechanics,mechanobiology,and mechanotherapy.Additionally,potential future development directions are prospectively analyzed to provide novel insights into wound care and strategies for preventing scar formation.
6.Influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts
Menglu WU ; Rui WANG ; Xinnan ZHENG ; Juan WU ; Lin HE ; Jiansheng DIAO ; Maoguo SHU ; Huicong DU
Chinese Journal of Burns 2025;41(4):355-363
Objective:To investigate the influence and mechanisms of metformin on the proliferation and apoptosis of human keloid fibroblasts (Fbs).Methods:This study was an experimental research. The keloid tissue was collected from 7 keloid patients (2 males and 5 females, aged 20-65 years, with a disease course of more than 1 year) who underwent keloid excision surgery at the Department of Plastic, Cosmetic and Maxillofacial Surgery of the First Affiliated Hospital of Xi'an Jiaotong University from September 2020 to September 2023. The primary Fbs were isolated and cultured, and cells from passages 3 to 6 were used for experiments. The cells were divided into control group and metformin group, and were cultured in complete medium. The medium for metformin group was supplemented with metformin at a final molarity of 60 mmol/L. The cell counting kit-8 was used to assess the proliferation activity of cells in two groups after 12 and 24 hours of culture, and the proliferation inhibition rate of cells in metformin group after 12 and 24 hours of culture was calculated, with a sample size of 6. The apoptosis detection kit was used to detect the apoptotic distribution of cells in control group after 0 hour (immediately) of culture and in metformin group after 12 and 24 hours of culture, with a sample size of 3. The cell cycle detection kit was used to detect the cycle distribution of cells in two groups after 12 and 24 hours of culture, with a sample size of 3. The eukaryotic mRNA sequencing was performed on suitable number of cells of two groups after 24 hours of culture, and the Kyoto encyclopedia of genes and genomes functional annotation analysis and functional enrichment analysis were performed after screening for differentially expressed genes (DEGs) with significantly differential expression between two groups. Western blotting was conducted to detect the protein expressions of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), and phosphorylated mammalian target of rapamycin (p-mTOR) in the PI3K/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway of cells in two groups after 24 hours of culture, with a sample size of 3.Results:After 12 and 24 hours of culture, the proliferation activity of cells in metformin group was significantly lower than that in control group (with t values of 4.70 and 24.02, respectively, P<0.05); the proliferation activity of cells in metformin group after 24 hours of culture was significantly lower than that after 12 hours of culture within the group ( t=4.73, P<0.05). Compared with that after 12 hours of culture within the group, the proliferation inhibition rate of cells in metformin group was significantly increased after 24 hours of culture ( t=5.29, P<0.05). Compared with that in control group after 0 hour of culture, the proportion of early apoptotic cells in metformin group was significantly increased (with t values of 6.62 and 4.58, respectively, P<0.05), and the proportion of early and late apoptotic cells was significantly increased after 12 and 24 hours of culture (with t values of 4.84 and 3.75, respectively, P<0.05). After 24 hours of culture, the proportion of late apoptotic cells in metformin group was significantly higher than that after 12 hours of culture within the group ( t=4.55, P<0.05). After 12 hours of culture, the proportion of S-phase cells in metformin group was significantly lower than that in control group ( t=5.90, P<0.05). After 24 hours of culture, compared with that in control group, the proportion of G0/G1-phase cells in metformin group was significantly increased ( t=5.36, P<0.05), while the proportion of G2/M-phase cells was significantly decreased ( t=17.63, P<0.05). The proportion of S-phase cells in metformin group after 24 hours of culture was significantly higher than that after 12 hours of culture within the group ( t=7.60, P<0.05). After 24 hours of culture, 4 814 DEGs with significantly differential expression were detected in the cells of metformin group compared with control group. The significantly upregulated and downregulated DEGs were mainly involved in biological functions related to signal transduction, cell growth and death, transport and catabolism, the endocrine system, the immune system, and cancer. The pathways that were significantly enriched with DEGs with significantly differential expression included the cell cycle and DNA replication, with the highest number of genes in the PI3K/Akt signaling pathway. After 24 hours of culture, the protein expressions of PI3K, p-Akt, and p-mTOR of cells in metformin group were 0.190±0.017, 0.170±0.017, and 0.247±0.005, respectively, which were significantly lower than 0.440±0.026, 0.300±0.060, and 0.547±0.025 in control group (with t values of 13.69, 3.61, and 20.12, respectively, P values all <0.05). Conclusions:Metformin can significantly inhibit the proliferation of human keloid Fbs through the PI3K/Akt/mTOR signaling pathway and effectively induce its apoptotic process, thereby exerting antifibrotic effects.
7.Effect of Cannabidiol on Pulmonary Fibrosis Analyzed by UPLC-Q-TOF-MS
Mengdi SUN ; Feiyu ZHANG ; Huicong YANG ; Yu WANG ; Pingping CHEN ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(12):185-193
ObjectiveTo investigate the mechanism of anti-pulmonary fibrosis of cannabidiol by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). MethodSD rats were randomly divided into blank group, model group, prednisone group(3.15 mg·kg-1) and cannabidiol low, medium and high dose groups(12, 36, 108 mg·kg-1), with 8 rats in each group. The rat model of pulmonary fibrosis was established by intratracheal injection of bleomycin(5 mg·kg-1), which was administered continuously for 28 days after successful modeling. The pathological changes of rat lung tissue were observed, and enzyme-linked immunosorbent assay(ELISA) was used to detect the expression levels of matrix metalloproteinase-7(MMP-7), type Ⅱ alveolar cell surface antigen(KL-6), pulmonary surfactant-associated protein A(SP-A) and SP-D in serum. The expression levels of type Ⅰ collagen(Col-Ⅰ) and fibronectin(FN) in lung tissues were detected by immunohistochemistry, and the expression of mucin 5 subtype AC(MUC5AC) was detected by immunofluorescence. UPLC-Q-TOF-MS was used to search for potential biomarkers and related metabolic pathways of cannabidiol in treating pulmonary fibrosis. ResultCompared with the blank group, there were a large number of inflammatory cell infiltration and continuous fibrosis lesions in the lung tissue of rats in the model group. Compared with the model group, the inflammatory infiltration and blue collagen deposition in the lung tissue of rats in the prednisone and cannabidiol groups were reduced. Compared with the blank group, the expressions of MMP-7, KL-6, SP-A and SP-D in serum of the model group were significantly increased(P<0.01), while the expressions of MMP-7, KL-6, SP-A and SP-D in the prednisone and cannabidiol high dose groups were significantly decreased by comparing with the model group(P<0.05, P<0.01). Compared with the blank group, the expression levels of Col-Ⅰ and FN in the lung tissues of the model group were significantly increased, and the fluorescence intensity of MUC5AC was significantly increased(P<0.01). Compared with the model group, the expression levels of Col-Ⅰ and FN in the lung tissues of the prednisone and cannabidiol high dose groups were significantly decreased(P<0.05, P<0.01), and the expression of MUC5AC was significantly decreased(P<0.01). Compared with the blank group, a total of 18 differential compounds were screened out in the model group, which could be used as potential biomarkers, and cannabidiol could call back 16 of them, mainly involving 4 metabolic pathways(linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, and niacin and niacinamide metabolism). Compared with the blank group, the relative contents of potential biomarkers arachidonic acid and linoleic acid were significantly increased in the model group(P<0.05, P<0.01), while the relative contents of 5,6-EET, L-tyrosine and niacinamide were significantly decreased(P<0.01). Compared with the model group, cannabidiol could significantly reduce the relative contents of arachidonic acid and linoleic acid, and significantly increase the relative contents of 5,6-EET, L-tyrosine and niacinamide(P<0.01). ConclusionCannabidiol has an intervention and remission effect on pulmonary fibrosis, and its mechanism may be related to linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, niacin and niacinamide metabolism.
8.Establishment and evaluation for nomogram model of rheumatoid arthritis with skeletal system involvement
Ziming WANG ; Guorui LIU ; Huicong ZHOU ; Xiaoling YAN ; Wei YU
Chinese Journal of Clinical Laboratory Science 2024;42(11):809-815
Objective To explore the risk factors of rheumatoid arthritis patients complicated with skeletal system involvement,and es-tablish a prediction model and evaluate its performance.Methods A total of 165 patients,who were initially diagnosed with rheuma-toid arthritis in Taikang Xianlin Drum Tower Hospital and Eastern Theater General Hospital from September 2015 to March 2021,were included in this study.The clinical and serological parameters of the patients with and without complicated skeletal system involvement were compared.The predictive factors with important significance in LASSO-Logistic analysis were used as the candidate variables,and a nomogram model was established based on the screened predictive factors.The discrimination and calibration of the model were evalu-ated with receiver operating curve(ROC)and the calibration curve,and the clinical effectiveness of the model was evaluated by clini-cal decision curve and clinical impact curve.Results The incidence of rheumatoid arthritis patients complicated with skeletal system involvement was 60%(99/165).Compared with the group of non-complicated with skeletal system involvement,statistically significant differences were found in age,course of disease,IgG4 level,IgG4/IgG ratio,rheumatoid arthritis(RF),antistreptolysin O(ASO),plateletcrit(PCT)and other parameters of laboratory tests in the complication group(P<0.05).The nomogram model of rheumatoid arthritis complicated with skeletal system involvement was established by five indicators,i.e.,age,course of disease,IgG4/IgG ratio,RF and PCT.The statistic of the model C was 0.848(95%CI:0.785 to 0.910).Calibration curves showed good predictive performance in internal validation,and the results of Hosmer-lemeshaw test indicated goodness of fit(χ2=7.562,P=0.477,>0.05).Both the clin-ical decision curve and clinical impact curve showed its clinical practicability.Conclusion The established simple and practical nomo-gram model exhibited good ability to identify rheumatoid arthritis patients complicated with skeletal system involvement,which may pro-vide reference basis for clinical diagnosis and treatment.
9.A clinical study on children with self-limited epilepsy with centrotemporal spikes and attention-deficit hyperactivity disorder comorbidity
Mengying WANG ; Huimin LI ; Yumei GENG ; Qing ZHOU ; Man WANG ; Huicong KANG
Chinese Journal of Neuromedicine 2024;23(6):552-559
Objective:To summarize the clinical and EEG characteristics of children with self-limited epilepsy with centrotemporal spikes (SeLECTS), and explore the risk factors for comorbid attention-deficit hyperactivity disorder (ADHD).Methods:Demographic and medical history data, seizure characteristics, EEG data, and treatment information of 122 children with SeLECTS admitted to Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from July 2020 to March 2024 were collected. Based on Swanson, Nolan and Pelham-IV Rating Scale (SNAP-IV) results, these patients were divided into comorbid ADHD group ( n=64) and non-ADHD group ( n=58); the clinical data and EEG characteristics (location and lateralization of discharges, synchronicity of bilateral discharges, period of discharges, spike wave frequency and special waveforms) of the patients between the 2 groups were compared to analyze the risk factors for comorbid ADHD. Results:(1) In 122 SeLECTS children, 70 (57.38%) were males and 52 (42.62%) were females; onset age was (7.50±1.98) years (3-12 years); 72.95% (89/122) patients had seizures only within 1 h after falling asleep, 9.84% (12/122) had seizures only 1-2 h before awakening in the morning, 9.84% (12/122) had seizures both after falling asleep and before awakening in the morning; duration of a single episode was (116.60±89.68) seconds (10-600 seconds). (2) Background activity in EEG showed no obvious abnormalities; the discharges were located in the central temporal region in 93.20% (96/103) patients and in the central temporal region and other brain regions (including frontal region, anterior head, bilateral occipital midline and bilateral cerebral hemispheres) in 6.80% (7/103) patients; among 69 patients whose overnight EEG recording was obtained, the spike frequency was 35.00 (20.67, 55.00) times/min (0.33-86.33 times/min). (3) Among 120 patients who accepted drug treatment, 87 (72.50%) received monotherapy, including valproic acid ( n=30, 34.48%), oxcarbazepine ( n=21, 24.14%), lacosamide ( n=17, 19.54%), levetiracetam ( n=17, 19.54%), perampanel ( n=1, 1.15%), and lamotrigine ( n=1, 1.15%); 33 (27.50%) received combination therapy with two or more drugs. (4) The comorbid ADHD group had statistically younger age of onset, longer duration of a single episode, higher proportion of seizures both after falling asleep and before awakening, and higher spike wave frequency in EEG than the non-ADHD group ( P<0.05). Conclusions:SeLECTS patients generally have onset age of 3-12 years, seizures within 1 h after falling asleep or 1-2 h before awakening, normal EEG background activity, and epileptiform discharges mostly located in the centrotemporal area, which are different from other types of epilepsy. SeLECTS patients with young age of onset, long duration of a single episode, seizures both after falling asleep and before awakening and high spike wave frequency in EEG trend to develop ADHD comorbidity.
10.Influence of three different nucleic acid extraction methods on 2019-nCoV
Huicong ZHOU ; Ziming WANG ; Yue HU ; Jie ZHANG ; Yanting ZHU ; Mei HUANG ; Xiaoling YAN ; Wei YU
Chinese Journal of Laboratory Medicine 2022;45(4):393-398
Objective:To study the difference in the extraction efficiency of the novel coronavirus (2019-nCoV) nucleic acid by using magnetic beads method, centrifugal column method and one-step method.Methods:On March 5, 2021, 10 throat swabs were collected from the staff working in the nucleic acid sampling room in Department of Clinical Laboratory, Affiliated Taikang Xianlin Drum Tower Hospital, Medical School of Nanjing University. The positive quality control samples were mixed into the swabs and used as mock positive samples. The RNA was extracted from simulated positive samples and their diluted samples by using magnetic beads method, centrifugation column method and one-step method. The purity ( A260/ A280 ratio) and concentration of the nucleic acid obtained were measured by micro-uv photometry, and fluorescence quantitative PCR was performed to compare the CT value and extraction efficiency. The three methods were used to extract the simulated weak positive specimens and to compare the difference of CT values after amplification. The measurement data that followed normal distribution were expressed by xˉ±s, the t test was used for comparing in the same group, and single factor analysis of variance was used for comparing among multiple groups. A P value smaller than 0.05 indicated a significant difference. Results:2019-nCoV nucleic acid extracted by magnetic bead method, centrifugal column method and one-step method could amplify positive results. There was no significant difference between the CT value of RNA amplification extracted by magnetic bead method and one-step method ( t=? 0.995 , P=0.376). The CT values of orf1ab gene amplified by centrifugal column method, magnetic bead method and one-step method were 29.28±0.06, 30.82±0.14 and 29.79±0.01 respectively ( F=11.196 , P=0.041). The CT values of E gene were 28.52±0.40, 27.33±0.78 and 27.38±0.13 respectively ( F=3.407, P=0.169). The CT values of N gene were 28.61±1.02, 27.24±0.20 and 27.25±0.47, respectively ( F=2.880 , P=0.020). The CT values of human genes extracted by centrifugal column method, magnetic bead method and one-step method were 19.68±0.36, 20.14±0.06 and 20.58±0.49 respectively, which was statistically significant ( F=4.904, P=0.048). The CT value of amplified human gene was affected by the dilution of human samples twice. The CT value of undiluted samples was smaller than that of diluted samples twice, with a difference of 2.95±0.22, which was statistically significant ( t=?3.025, P=0.039). The extraction time of one-step method, magnetic bead method and centrifugal column method were (15.00±1.50), (20.00±1.50) and (40.00±5.5) min respectively, and the difference was statistically significant ( F=688 , P=0.027). Conclusions:Magnetic bead method, centrifugal column method and one-step method can be used to extract 2019-nCoV nucleic acid, for the centrifugal column method has a higher extraction efficiency than the magnetic bead method and the one-step method. The one-step method is the fastest, followed by the magnetic bead method and the centrifugal column method. A large number of clinical samples can be processed using the magnetic bead method and one-step method. One-step rapid nucleic acid test can also be performed on samples from emergency and fever clinics. It is not recommended to dilute specimens for testing. In order to improve the detection rate, extracting RNA from highly suspected samples with negative initial nucleic acid test by centrifugal column method is suggested.

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