1.Detection of H5N6 and H9N2 subtypes of avian influenza viruses with reverse transcription-recombinase polymerase amplification combined with CRISPR-Cas13a
Jing-jing WU ; Yu-wei WENG ; Zhi-miao HUANG ; Hong-bin CHEN
Chinese Journal of Zoonoses 2025;41(3):235-242
The aim of this study was to establish a rapid,highly sensitive,and specific nucleic acid detection method for the H5N6 and H9N2 avian influenza virus(AIV)subtypes by using reverse transcription-recombinase polymerase amplification(RT-RPA)combined with clustered regularly interspaced short palindromic repeats(CRISPR)-Cas13a proteins.The conserved regions were selected to design specific RT-RPA primers and crRNA sequences of H5,H6,H9,and N2 genes.RT-RPA tech-nology combined with CRISPR-Cas13a detection was used to evaluate the sensitivity and specificity of AIV nucleic acid detec-tion.The detection was performed on avian influenza environmental samples and compared with the results of fluorescence quantitative RT-PCR,to evaluate the effectiveness of the RT-RPA technology combined with CRISPR-Cas13a.AIV H5,H9,and N2 subtypes were detected with a sensitivity as high as 1 copy/μL,and AIV N6 subtypes were detected with a sensitivity of 10 copies/μL.Plasmid samples with differing copy numbers showed fluorescence under blue LED transillumination.The four AIV subtypes showed high specificity and did not cross-react with the other AIV subtypes.The detection of avian influenza ex-ternal environmental samples containing AIV H5,N6,and H9 subtypes was consistent with the results of fluorescence quanti-tative RT-PCR,with 100%accuracy.For AIV N2 subtypes,one additional negative sample was detected with 97.9%accuracy.The established RT-RPA technology combined with CRISPR-Cas13a detection enabled sensitive,specific visual detection of AIV H5N6 and H9N2 subtypes.This study provides a new nucleic acid detection method for AIV surveillance and subtype clas-sification.
2.Typical failure treatment of large-aperture 16-slice spiral Siemens SOMATOM Sensation Open CT
Yu-kun ZHU ; Shi-dong CHENG ; Ming YANG ; Fei WENG ; Jing TIAN ; Chen LIANG
Chinese Medical Equipment Journal 2025;46(11):112-114
Three typical failures of large-aperture 16-slice spiral Siemens SOMATOM Sensation Open CT were introduced in terms of phenomenon,cause and treatment method.References were provided for medical engineers to treat similar failures.
3.Typical failure treatment of large-aperture 16-slice spiral Siemens SOMATOM Sensation Open CT
Yu-kun ZHU ; Shi-dong CHENG ; Ming YANG ; Fei WENG ; Jing TIAN ; Chen LIANG
Chinese Medical Equipment Journal 2025;46(11):112-114
Three typical failures of large-aperture 16-slice spiral Siemens SOMATOM Sensation Open CT were introduced in terms of phenomenon,cause and treatment method.References were provided for medical engineers to treat similar failures.
4.Detection of H5N6 and H9N2 subtypes of avian influenza viruses with reverse transcription-recombinase polymerase amplification combined with CRISPR-Cas13a
Jing-jing WU ; Yu-wei WENG ; Zhi-miao HUANG ; Hong-bin CHEN
Chinese Journal of Zoonoses 2025;41(3):235-242
The aim of this study was to establish a rapid,highly sensitive,and specific nucleic acid detection method for the H5N6 and H9N2 avian influenza virus(AIV)subtypes by using reverse transcription-recombinase polymerase amplification(RT-RPA)combined with clustered regularly interspaced short palindromic repeats(CRISPR)-Cas13a proteins.The conserved regions were selected to design specific RT-RPA primers and crRNA sequences of H5,H6,H9,and N2 genes.RT-RPA tech-nology combined with CRISPR-Cas13a detection was used to evaluate the sensitivity and specificity of AIV nucleic acid detec-tion.The detection was performed on avian influenza environmental samples and compared with the results of fluorescence quantitative RT-PCR,to evaluate the effectiveness of the RT-RPA technology combined with CRISPR-Cas13a.AIV H5,H9,and N2 subtypes were detected with a sensitivity as high as 1 copy/μL,and AIV N6 subtypes were detected with a sensitivity of 10 copies/μL.Plasmid samples with differing copy numbers showed fluorescence under blue LED transillumination.The four AIV subtypes showed high specificity and did not cross-react with the other AIV subtypes.The detection of avian influenza ex-ternal environmental samples containing AIV H5,N6,and H9 subtypes was consistent with the results of fluorescence quanti-tative RT-PCR,with 100%accuracy.For AIV N2 subtypes,one additional negative sample was detected with 97.9%accuracy.The established RT-RPA technology combined with CRISPR-Cas13a detection enabled sensitive,specific visual detection of AIV H5N6 and H9N2 subtypes.This study provides a new nucleic acid detection method for AIV surveillance and subtype clas-sification.
5.The application of family empowerment model on the primary caregivers of first-episode stroke dysphagia patients
Hong YU ; Jing DU ; Qian XU ; Mingming XU ; Xiangge FAN ; Fan ZHANG ; Xueyun WENG ; Xiaoming MA ; Yanhua HOU ; Linqing LI
Chinese Journal of Practical Nursing 2024;40(4):263-271
Objective:To explore the effect of family empowerment model on the improvement of swallowing care ability and care preparedness of primary caregivers of first-episode stroke dysphagia patients, further to explore its impact on patients′s wallowing function and life quality.Methods:This study was a randomized controlled study. From January 2021 to December 2022, 80 main caregivers of patients with dysphagia caused by manual stroke admitted to the Department of Acupuncture and Moxibustion, Shenzhen Hospital of Traditional Chinese Medicine were selected as the research objects, and 40 cases in the control group and 40 cases in the observation group were selected by random number table method. The control group were treated with conventional nursing care of first-episode stroke dysphagia patients in the acupuncture and moxibustion Department. On the basis of the conventional care in the control group, the observation group were treated with family empowerment model intervention for 14 days and was followed up for 28 days. Primary caregivers′ swallowing care ability, Caregiver Preparedness Scale (CPS), patients′ swallowing function rate, Swallowing Related Quality of Life (SWALQOL) were used to evaluate the effects before intervention and at the end of intervention.Results:There were 18 males and 19 females primary caregivers in the control group, aged (55.61 ± 7.43) years old. There were 18 males and 21 females primary caregivers in the observation group, aged (58.23 ± 8.22) years old. The swallowing care ability score showed a statistically significant difference between the observation group (143.47 ± 3.96) and the control group (107.74 ± 1.43) ( t=-26.76, P<0.05). After intervention, the caregiver preparedness scale was (26.11 ± 3.81) in the observation group, and (18.35 ± 4.54) in the control group, and the difference was statistically significant ( t=-4.11, P<0.05).The patients′ swallowing function rate and SWALQOL score were respectively 97.44% (38/39) and (91.41 ± 8.08) points in the observation group, and 72.97% (27/37) and (80.33 ± 4.21) points in the control group, and the difference was both statistically significant ( χ2=10.76, t=-2.54, both P<0.05). Conclusions:The implementation of family empowerment model could enhance the swallowing care ability and care preparedness of primary caregivers of the first-episode stroke dysphagia patients, which could further improve patients′ swallowing function and life quality.
6.Mechanism of dorsal root ganglion SERT in electroacupuncture regulation of P2X3 receptor-mediated visceral hypersensitivity in IBS rats
Rong HUANG ; Jing CHAI ; Yun ZHOU ; Yu QIAO ; Zhijun WENG ; Huangan WU ; Huirong LIU ; Lu ZHU ; Jindan MA ; Yi ZHU ; Fang ZHANG ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(6):435-446
Objective:To investigate the role of serotonin reuptake transporter(SERT)and P2X3 receptor of dorsal root ganglion(DRG)in regulating visceral hypersensitivity of rats with irritable bowel syndrome(IBS)by electroacupuncture(EA). Methods:Male Sprague-Dawley and SERT-/-rats were subjected to preparing IBS visceral hypersensitivity models with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.Three weeks post-modeling,interventions including EA,intrathecal injection,and EA plus intrathecal injection were applied,respectively.Hematoxylin-eosin staining and abdominal withdrawal reflex(AWR)score were used to confirm the successful establishment of the IBS model.AWR score,whole-cell patch clamp technique,and Western blotting assay were used to evaluate the changes in visceral pain sensitivity,electrophysiological properties of DRG neurons,and the expression of DRG P2X3 receptor and SERT in IBS rats. Results:Compared to the model group,the AWR score in the EA group decreased significantly(P<0.05),the resting membrane potential(P<0.05)and the number of action potentials(P<0.05)of DRG neurons reduced,and the baseline intensity increased(P<0.05);additionally,the expression of P2X3 receptor in DRG decreased(P<0.01),and the SERT expression increased(P<0.05).Compared to the P2X3 receptor agonist group,the SERT protein expression in DRG was higher in the EA group.In SERT-/-rats,the P2X3 receptor expression in DRG increased in the EA group compared to the model group(P<0.01). Conclusion:EA modulates the electrophysiological characteristics of intestinal primary sensory neurons by regulating the expression of SERT and P2X3 receptor in DRG of IBS rats.This modulation may contribute to the mechanism by which EA alleviates peripheral sensitization of visceral pain in IBS rats.
7.Analysis of the efficacy and influencing factors of sodium channel blockers in the treatment of focal epilepsy in infants under 6 months of age.
Yu Ping MA ; Jie DENG ; Zheng Ran FU ; Chun Hong CHEN ; Xiao Hui WANG ; Xu WANG ; Jing Wen WENG ; Yan Hua SHEN
Chinese Journal of Pediatrics 2023;61(11):983-988
Objective: To analyze the efficacy and safety of the sodium channel blockers (SCB) antiseizure medication in the treatment of focal epilepsy in infants under 6 months of age. Methods: This was a case series study. Infants with focal epilepsy with onset within 6 months of age and treated with SCB attending the Department of Neurology of Beijing Children's Hospital from June 2016 to April 2022 were collected. The clinical data, auxiliary examinations, SCB application, efficacy, adverse reactions, and prognosis were analyzed retrospectively. Patients were grouped according to type of seizure and epileptic syndrome, age of onset and etiology. Chi square test and Fisher exact test were used to analyze the differences between groups statistically. Results: A total of 118 infants were enrolled, 65 males and 53 females, with an age of epilepsy onset of 56 (4, 114) days. Developmental and epileptic encephalopathy was diagnosed in 60 infants, 39 had self-limited neonatal and (or) infantile epilepsy, and 19 had non-syndromic focal epilepsy. Application of SCB: 106 used oxcarbazepine, 2 used lacosamide, 9 switched from oxcarbazepine to lacosamide or a combination of 2 SCB, and 1 used oxcarbazepine, lacosamide, and lamotrigine successively; oxcarbazepine was the first choice in 46 cases. The age at which SCB was applied was 103 (53, 144) days. The children were followed up for 6 months to 6 years. SCB was effective in 89 cases (75.4%), including 70 cases (59.3%) who achieved seizure freedom. The seizure-free rate was higher in the focal epilepsy only group than in the group with other seizure types (64.4% (65/101) vs. 4/17, χ²=9.99, P<0.05). The responder and seizure-free rates were all higher in the group with the onset age of >3-6 months than the group >1-3 months (84.4% (38/45) vs. 62.5% (20/32), 73.3% (33/45) vs. 46.9% (15/32), χ²=4.85 and 5.58, both P<0.05). With the exception of variants in the PRRT2 gene, those with variants in sodium or potassium channels had higher responder and seizure-free rates than those with variants in other genes(86.2% (25/29) vs. 45.5% (10/22), 62.1% (18/29) vs. 22.7% (5/22), χ²=9.65 and 7.82,both P<0.05). The most common adverse event was transient hyponatremia, which happened in 66 cases (55.9%). There were 9 cases of rash, which subsided in 6 cases after discontinuing oxcarbazepine and switching to lacosamide, and 7 cases of electrocardiogram abnormalities, which improved after withdrawing oxcarbazepine and changing to lacosamide in 1 case. Conclusion: SCB are effective and tolerable in the treatment of focal epilepsy in infants under 6 months of age, with better efficacy in patients with genetic variants of the sodium or potassium channel, focal seizures only, and seizure onset >3-6 months of age.
Child
;
Female
;
Male
;
Infant, Newborn
;
Humans
;
Infant
;
Sodium Channel Blockers/adverse effects*
;
Oxcarbazepine
;
Lacosamide
;
Retrospective Studies
;
Epilepsies, Partial/drug therapy*
;
Seizures
;
Sodium
;
Anticonvulsants/adverse effects*
8.Effect of patent foramen ovale on perioperative complications and survival rate in pediatric patients undergoing living donor liver transplantation: a retrospective study
Yuli WU ; Yiqi WENG ; Yongle JING ; Tianying LI ; Jianjiang WU ; Mingwei SHENG ; Lili JIA ; Weihua LIU ; Wenli YU
Chinese Journal of Anesthesiology 2023;43(8):931-936
Objective:To evaluate the effect of patent foramen ovale (PFO) on the perioperative complications and survival rate in pediatric patients undergoing living donor liver transplantation.Methods:The medical records from pediatric patients of either sex with biliary atresia, aged<18 yr, who underwent living donor liver transplantation from January 2020 to January 2022, were retrospectively collected. The pediatric patients were divided into PFO group and non-PFO group according to the results of echocardiography before operation. The postreperfusion syndrome, acute lung injury, acute kidney injury, postoperative delirium and 1-year survival rate were recorded.Results:There was no significant difference in the incidence of postreperfusion syndrome, acute lung injury, acute kidney injury, postoperative delirium and one-year survival rate between PFO group and non-PFO group ( P>0.05). Conclusions:PFO has no obvious effect on the incidece of intraoperative and early postoperative complications and 1-year survival rate in pediatric patients undergoing living donor liver transplantation.
9.Risk factors for postreperfusion syndrome during living donor liver transplantation in pediatric patients with biliary atresia
Yuli WU ; Yiqi WENG ; Yongle JING ; Tianying LI ; Lu CHE ; Mingwei SHENG ; Lili JIA ; Weihua LIU ; Wenli YU
Chinese Journal of Anesthesiology 2023;43(9):1054-1058
Objective:To identify the risk factors for postreperfusion syndrome (PRS) during living donor liver transplantation in pediatric patients with biliary atresia.Methods:The clinical data from pediatric patients who underwent living donor liver transplantation from January 2020 to December 2021 in our hospital were retrospectively analyzed. The clinical data included: (1) general information of the pediatric patients such as age, gender, height and body weight; (2) preoperative data such as left ventricular ejection fraction, pediatric end-stage liver disease score, serum aminotransferase, aspartate aminotransferase, total bilirubin, International Normalised Ratio and creatinine concentrations, and whole blood Hb concentration; (3) intraoperative data such as vital signs and blood gas analysis parameters immediate before reperfusion, time of anhepatic phase, donor liver cold ischemia time, transplanted liver quality, time of surgery, anesthesia time, volume of urine, blood loss, amount of blood transfused, and amount of fresh frozen plasma transfused. The pediatric patients were divided into PRS group and non-PRS group according to whether intraoperative PRS occurred. Risk factors for PRS were analyzed using binary logistic regression analysis.Results:A total of 304 pediatric patients were finally enrolled, with 132 cases in PRS group and 172 cases in non-PRS group. The incidence of PRS was 43.4%. The results of logistic regression analysis showed that prolonged liver graft cold ischemic time ( OR=1.031, 95% confidence interval 1.021-1.042, P<0.001) and body temperature <36 ℃ immediately before reperfusion ( OR=3.095, 95% confidence interval 1.656-5.785, P<0.001) were risk factors for PRS. Conclusions:Body temperature immediately before reperfusion<36.0 ℃ and prolonged liver graft cold ischemic time are risk factors for PRS during living donor liver transplantation in pediatric patients with biliary atresia.
10.Effect of Qinggan Xiefei Fang on increasing susceptibility of influenza virus caused by "liver fire invading lung"
Yu-hui LU ; Shu-hua OUYANG ; Jing-yu WENG ; Pei LIU ; Xin-xing CHEN ; Kurihara HIROSHI ; Yi-fang LI ; Rong-rong HE
Acta Pharmaceutica Sinica 2022;57(6):1649-1656
In this study, chronic emotional stress-induced H1N1 influenza susceptibility model was employed to simulate the states of "emotional stagnation" and "liver fire invading lung", and the protective effect of Qinggan Xiefei Fang on viral pneumonia was investigated. Survival rate and morbidity rate of mice were observed within 21 days after H1N1 infection, the symptoms of viral pneumonia and the level of phospholipid peroxidation were detected in lungs of mice after 6-day infection. The experimental results showed that Qinggan Xiefei Fang could alleviate the decline of survival rate and morbidity rate of mice caused by chronic constraint stress loaded with H1N1, inhibit the replication of H1N1 and the production of inflammatory factors, reduce the level of phospholipid peroxidation, and improve the symptoms of pneumonia in mice. The results also showed that compound-target network of Qinggan Xiefei Fang contained 171 compounds and 260 corresponding targets involved in the signaling pathway of oxidative stress, inflammation and immunity. All the above results indicate that Qinggan Xiefei Fang protecting influenza virus pneumonia was related to the regulation of oxidative stress. The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Jinan University, in compliance with the Institutional Animal Care Guidelines.

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