1.Establishment of real-time fluorescence quantitative PCR method for detecting the N subgenome of SARS-CoV-2
Taoli HAN ; Zhi ZHANG ; Jiaxin ZHAO ; Pan LU ; Yang JIAO ; Jianhong ZHAO ; Yan GAO ; Shiyao ZHANG ; Kuankuan LIU ; Yujie LIU ; Ru FAN ; Wenjing LI ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):96-101
Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.
2.A real world study on the effectiveness and safety of deu-cravacitinib in the treatment of moderate-to-severe plaque psoriasis in China.
Yi ZHANG ; Tongyao CHEN ; Wenjing PAN ; Zhiming LI
Journal of Zhejiang University. Medical sciences 2025;54(5):653-660
OBJECTIVES:
To evaluate the effectiveness and safety of deucravacitinib in Chinese patients with moderate-to-severe plaque psoriasis.
METHODS:
This retrospective study included 41 patients with moderate-to-severe plaque psoriasis treated with deu-cravacitinib 6 mg once daily for 16 weeks at the First Affiliated Hospital of Wenzhou Medical University between January and September 2024. Effectiveness was assessed by the psoriasis area and severity index (PASI), static physician's global assessment (sPGA), palmoplantar psoriasis area and severity index (PPASI), modified nail psoriasis severity index (mNAPSI), and dermatology life quality index (DLQI) at baseline, 4, 8, 12 and 16 weeks after treatment. Adverse events during treatment were recorded. Laboratory parameters, including complete blood count, liver and kidney function, electrolytes, and lipids, were assessed at baseline, 8, 16 weeks after treatment to evaluate safety. Univariate and multivariate logistic regression analyses were performed to explore factors associated with achieving PASI75 at week 16, using baseline characteristics as independent variables.
RESULTS:
Significant reductions from baseline in PASI and DLQI scores were observed from week 4 through week 16 (all P<0.01). Overall response rates for PASI75, PASI90, PASI100, sPGA 0 or 1 grade, and DLQI 0 or 1 point increased progressively over the treatment period. 75, 90, and 100 refer to a score reduction of at least 75%, at least 90%, and 100%, respectively, from baseline. Response rates of PASI75, PASI90, PASI100 for the scalp, limbs, and trunk, PPASI75, PPASI90, PPASI100 for palmoplantar lesions, and mNAPSI75, mNAPSI90 for nail lesions increased progressively over time but with different trends. Scalp lesions improved most markedly from week 4, followed by the limbs, whereas improvements in trunk and palmoplantar lesions were relatively slower. Nail lesions responded more slowly, with only 20% of patients achieving marked improve-ment at week 16. Deucravacitinib demonstrated good tolerability and compatibility with concomitant medications. No severe adverse events were reported, indicating a favorable safety profile. Multivariate logistic regression analysis revealed no significant association between the achievement of PASI75 response at week 16 and patient age, body mass index, disease duration, or baseline PASI, sPGA, or DLQI scores (all P>0.05).
CONCLUSIONS
In this real world study of Chinese patients with moderate-to-severe plaque psoriasis, deucravacitinib demonstrated favorable effectiveness and safety over 16 weeks of treatment.
Humans
;
Psoriasis/drug therapy*
;
Retrospective Studies
;
Male
;
Female
;
Middle Aged
;
China
;
Adult
;
Treatment Outcome
;
Severity of Illness Index
;
Quality of Life
;
Aged
3.Advances in cancer stem cell markers and their targeting aptamers.
Shangyang PAN ; Wenjing ZHANG ; Xiaoyang CHEN ; Yan LIU ; Ruolan CHEN ; Shuyue MENG ; Zhao YANG
Chinese Journal of Biotechnology 2025;41(8):3008-3020
Cancer ranks as the second leading cause of death globally and has surpassed cardiovascular diseases to become the primary cause of mortality in developed countries. Cancer stem cells (CSCs), which play crucial roles in cancer recurrence, metastasis, and drug resistance, have attracted significant attention in targeted therapeutic strategies. Aptamers, with unique three-dimensional structures capable of specifically recognizing the surface markers of CSCs, show promising potential in targeted drug delivery systems. Compared with conventional antibodies, aptamers are praised for small molecular weights, low production costs, and easy chemical modification. This review systematically summarizes recent advances in aptamer research targeting the surface markers of CSCs, with particular emphasis on aptamer-drug conjugate systems targeting the markers including EpCAM, CD133, CD44, and ABCG2. Both in vitro cellular studies and in vivo animal models have demonstrated the definite anti-cancer efficacy of aptamer-based drug delivery systems, which are of great significance to develop novel therapeutic strategies and improving the therapeutic effects of CSC-targeted treatment. Thus, aptamer-based drug delivery system has broad application prospects in the field of precise cancer treatment.
Humans
;
Neoplastic Stem Cells/metabolism*
;
Aptamers, Nucleotide/therapeutic use*
;
Drug Delivery Systems/methods*
;
Neoplasms/drug therapy*
;
Biomarkers, Tumor/metabolism*
;
Animals
;
Epithelial Cell Adhesion Molecule
;
AC133 Antigen
;
Hyaluronan Receptors
4.Construction and optimization of inpatient medical quality evaluation index system in public hospitals based on life cycle theory
Xinxiang PAN ; Zhongliang BAI ; Wenjing CHEN ; Wenjie FU ; Huan ZHOU ; Hongju WANG
Journal of Shenyang Medical College 2025;27(1):20-25
Objective:To construct a medical quality evaluation index system for inpatients in public hospitals based on the life cycle theory,starting from the entire process of medical treatment.Method:A comprehensive study was conducted on the screening of medical quality evaluation indicators for hospitalized patients using literature analysis,key informant interviews,and expert inquiry methods.Results:The effective recovery rate of the two rounds of expert consultation was 100%,with high enthusiasm from the experts.The authority coefficient was above 0.7,and Kendall's W coordination coefficient was 0.267(P<0.05).The evaluation indicators for medical quality of hospitalized patients,including 3 primary indicators,10 secondary indicators,and 51 tertiary indicators,were determined.Conclusions:The medical quality evaluation index system for inpatients in public hospitals based on the life cycle theory has certain scientificity and reliability.However,the weight analysis of the index system has not been carried out and is still in the theoretical exploration stage.Further empirical research is needed for verification and improvement.
5.Construction and optimization of inpatient medical quality evaluation index system in public hospitals based on life cycle theory
Xinxiang PAN ; Zhongliang BAI ; Wenjing CHEN ; Wenjie FU ; Huan ZHOU ; Hongju WANG
Journal of Shenyang Medical College 2025;27(1):20-25
Objective:To construct a medical quality evaluation index system for inpatients in public hospitals based on the life cycle theory,starting from the entire process of medical treatment.Method:A comprehensive study was conducted on the screening of medical quality evaluation indicators for hospitalized patients using literature analysis,key informant interviews,and expert inquiry methods.Results:The effective recovery rate of the two rounds of expert consultation was 100%,with high enthusiasm from the experts.The authority coefficient was above 0.7,and Kendall's W coordination coefficient was 0.267(P<0.05).The evaluation indicators for medical quality of hospitalized patients,including 3 primary indicators,10 secondary indicators,and 51 tertiary indicators,were determined.Conclusions:The medical quality evaluation index system for inpatients in public hospitals based on the life cycle theory has certain scientificity and reliability.However,the weight analysis of the index system has not been carried out and is still in the theoretical exploration stage.Further empirical research is needed for verification and improvement.
6.Establishment of real-time fluorescence quantitative PCR method for detecting the N subgenome of SARS-CoV-2
Taoli HAN ; Zhi ZHANG ; Jiaxin ZHAO ; Pan LU ; Yang JIAO ; Jianhong ZHAO ; Yan GAO ; Shiyao ZHANG ; Kuankuan LIU ; Yujie LIU ; Ru FAN ; Wenjing LI ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):96-101
Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.
7.In vitro study of immunocompatibility of humanized genetically modified pig erythrocytes with human serum
Leijia CHEN ; Mengyi CUI ; Xiangyu SONG ; Kai WANG ; Zhibo JIA ; Liupu YANG ; Yanghui DONG ; Haochen ZUO ; Jiaxiang DU ; Dengke PAN ; Wenjing XU ; Hongbo REN ; Yaqun ZHAO ; Jiang PENG
Organ Transplantation 2024;15(3):415-421
Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.
8.Analysis of goitrogenic effect of goitrogen in food
Haowen PAN ; Honglei XIE ; Xin HOU ; Meng ZHAO ; Wenjing CHE ; Jia LI ; Yue SU ; Lanchun LIU ; Zexu ZHANG ; Zongyu YUE ; Peng LIU
Chinese Journal of Endemiology 2024;43(1):77-81
Goiter is a kind of non-inflammatory and non-neoplastic hyperplasia and enlargement. Many studies have shown that substances such as thiocyanates and isothiocyanates can prevent the development of a variety of tumors. However, some studies have also found that such substances can lead to goiter. In this article, relevant information on common goitrogen in food are collected to explore their mechanism of action, laying a foundation for guiding residents to maintain a healthy and balanced diet.
9.Analysis of the efficacy of vocal cord botulinum toxin injection for refractory laryngeal contact granuloma
Jiasen WANG ; Jinrang LI ; Yufei PAN ; Zhi LIU ; Chun ZHANG ; Wenjing WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(10):1048-1053
Objective:To observe the efficacy of vocal cord botulinum toxin type A injection in the treatment of refractory laryngeal contact granuloma and to analyze the factors affecting the curative effect.Methods:Fifty-two patients with refractory laryngeal contact granuloma who received vocal cord botulinum toxin type A injection under topical anesthesia from May 2021 to May 2023 were analyzed retrospectively. Among them, 51 were males and 1 was female, aged 22-66 (48.98±8.87)years old. All patients were followed up for a minimum of 12 months. Outcome measures in terms of patient cure rate, total effective rate, complications and recurrence rate were calculated. The median [ M ( Q1, Q3)] was used to represent non-normally distributed measurement data, and Logistic regression analysis was used to determine independent risk factors affecting efficacy. Results:The cure rate of 52 patients with refractory laryngeal contact granuloma treated by vocal cord botulinum toxin type A injection was 78.8% (41/52), and the total effective rate (including cure, marked and effective) was 90.4% (47/52). The median number of injections was 1[1,2]. Following a single injection, the cure rate was 69.2% (36/52), and the median treatment duration for cured patients was 3 [3,3] months. Hoarseness occurred in 88.5% (46/52) of patients, with recovery within 3 months in all cases. Additionally, 21.2% (11/52) of the patients experienced cough, sore throat, dyspnea, all of whom recovered within 3 months. One patient among the 41 cured cases was lost to follow-up, and the recurrence rate at the 12th month was 17.5% (7/40). Multivariate regression analysis indicated that age, granuloma size, history of PPI treatment, previous corticosteroid injections, prior surgical excision, pharyngeal reflux, chronic cough, pharyngeal reflux, chronic cough and vocal overuse were not independent risk factors for cure and recurrence.Conclusion:Vocal cord botulinum toxin type A injection is an alternative for the treatment of refractory laryngeal contact granuloma, which offers benefits such ashigh cure rate, short treatment cycle and less injection times.
10.Status quo of postoperative survival quality in 343 cases of heart valve replacement in Three Gorges Reservoir area and its influencing factors analysis
Jianrong ZHANG ; Jiqin ZHANG ; Dan CHENG ; Wenjing ZHANG ; Ailin DENG ; Min WANG ; Maolu WANG ; Yan ZHAO ; Deqiong DENG ; Pan WEI
Chongqing Medicine 2024;53(8):1248-1253
Objective To evaluate the postoperative survival quality in the patients with heart valve re-placement (HVR) in Three Gorges Reservoir area,and to analyze its main influencing factors.Methods A to-tal of 343 valvular heart disease patients from Three Gorges Reservoir area who received HVR treatment for the first time in this hospital from January 2019 to December 2021 were selected by the convenience sampling method.The general data questionnaire and the MOS 36-item short form health survey (SF-36) were adopted to conduct the survey.The main influencing factors affecting the survival quality were analyzed.Results Af-ter HVR,the physical components summary (PCS) score of SF-36 was 238.0±73.6,and the mental compo-nents summary (MCS) score was 254.8±83.6,and the scores in each dimension were significantly lower than those of the Chinese norm (P<0.05).The multiple stepwise linear regression analysis results showed that the age,gender,place of residence,education level,postoperative time,complications and readmission were the influencing factors of PCS and MCS scores (P<0.05).Conclusion The survival quality of the patients af-ter HVR is different from that of healthy population.The targeted intervention could be carried out according to the influencing factors of the survival quality of the patients,so as to improve their survival quality.

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