1.Construction of luminescent bacteriophage using CRISPR technology and its application in Escherichia coli indentification
Minwei LI ; Jing YAN ; Hangyi LI ; Zhiyun HAO ; Zhong NI ; Zhaoyang HU ; Xiaorong WANG ; Menghan XU ; Chi WANG ; Ruibing LI ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2024;47(4):437-443
Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.
2.Plantar Pressure Analysis after Medial Opening Wedge High Tibial Osteotomy Treatment of Medial Single Compartmental Knee Osteoarthritis
Zhaoyang WANG ; Zhong QING ; Liqiang ZHI ; Qiang ZAN ; Hua SHAN ; Le SHANG ; Jianbing MA ; Chao XU
Journal of Medical Biomechanics 2023;38(2):E310-E316
Objective To analyze the plantar pressure distribution of knee osteoarthritis ( KOA) patients after medial opening wedge high tibial osteotomy ( MOWHTO), so as to provide biomechanical references for the surgical treatment and rehabilitation of patients. Methods A total of 31 patients with medial single compartmental KOA after unilateral MOWHTO treatment were selected as the experimental group, and 35 healthy subjects at same age were selected as the control group. The Pedomedic 40 pressure measuring system was used to test dynamic plantar pressure. By comparing the maximum pressure ( pmax ), force-time integral ( FTI) and contact area (CA) of different plantar zones between the experimental group (operative side and unoperated side) and the control group during walking, the changes of plantar pressure in patients with medial single compartmental KOA after MOWHTO were evaluated. Results Compared with the unoperated side and the control group, the CA and FTI of the 1st metatarsal head (MH1) were higher (P<0. 05), the CA of the 4th metatarsal head (MH4)was smaller (P<0. 001), the pmax and FTI of the 5th metatarsal head (MH5) were smaller (P<0. 05), the CA of the lateral middle foot (MF-L) was smaller (P<0. 001), and the CA of the medial rear foot (RF-M) was larger (P<0. 05). Compared with the control group, the pmax of MH1 and MH2 was smaller (P<0. 05), the CA and FTI of MH5 were larger (P<0. 05), the pmax of MF-L was larger (P<0. 001), and the FTI of lateral rear foot (RF-L) was larger (P<0. 05). Conclusions Compared with healthy people, patients with medial single compartmental KOA have abnormal plantar pressure residual after MOWHTO. In clinical practice, targeted intensive rehabilitation therapy is necessary to restore the normal plantar distributions of patients.
3.Application of case-based learning based on WeChat Rain Classroom in large class teaching
Aimei LIU ; Hongmei LI ; Zhaoyang ZHONG
Chinese Journal of Medical Education Research 2022;21(3):300-304
Objective:To explore the application effects of case-based learning (CBL) based on Rain Classroom in surgical nursing teaching of large class.Methods:A total of 357 nursing undergraduates of Batch 2017 in Fenyang College of Shanxi Medical University were selected as the research objects and randomly divided into experimental group (176 cases) and control group (181 cases). The experimental group adopted the CBL based on Rain Classroom, while the control group adopted the traditional multimedia classroom CBL mode. After the teaching, the teaching effect was evaluated by interviewing, comparing the scores of theoretical examination and questionnaire survey. SPSS 19.0 was performed for t test. Results:The scores of memory type, comprehension type and comprehensive application type questions in the experimental group were significantly higher than those in the control group ( P<0.01). The scores of the experimental group were significantly higher than those of the control group in stimulating thirst for knowledge, learning initiative, classroom learning efficiency, memory of knowledge points, problem analysis, clinical thinking and team cooperation ability ( P<0.01). The interview information suggested that the interest of pre-study materials, the control of discussion links and the network effect were the problems affecting the classroom effect of the experimental group, while the control group had the problems of poor learning autonomy and enthusiasm. Conclusion:The CBL based on Rain Classroom can mobilize the initiative and participation of students in large class teaching, improve the learning effect and cultivate comprehensive ability, which is worthy of promotion.
4.Exploration of the talent training mode of biology majors driven by high-quality employments in higher education--a case study of Huzhou University.
Zhanqi WANG ; Xiaoyin WU ; Xueting ZHONG ; Hua HUANG ; Xiaomei WANG ; Zhaoyang TANG ; Jie YANG ; Liqin ZHANG
Chinese Journal of Biotechnology 2022;38(12):4827-4837
In recent years, driven by the support of national policies and societal needs for employments, talents in biology majors have been growing rapidly. To foster high-calibre biology talents for the society in the context of the "double world-class initiative" in higher education, this study analyzed the opinion of biology undergraduates in Huzhou University on employment and their professional recognition of biology majors. The aim of this study was to propose a high-quality employments-driven talent training mode for undergraduates in biology majors, so as to serve as a reference for the reform in training modes of other relevant majors.
Humans
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Universities
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Students
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Biology/education*
5.Efficacy of bicalutamide in the treatment of prostate cancer
Chinese Journal of Primary Medicine and Pharmacy 2020;27(11):1290-1293
Objective:To explore the therapeutic effect of bicalutamide on prostate cancer.Methods:Ninety-five patients with prostate cancer in the First People's Hospital of Jiande were selected from January 2014 to January 2019.All 95 patients received blood samples before treatment were selected as case group, and 85 healthy subjects were selected as control group.The clinical symptoms of the patients before treatment and after treatment with bicalutamide for 3, 6, 9, 12 months were recorded.The blood samples were taken as the treatment group at 12 months after treatment.The effect of bicalutamide on the expression of miRNA-125b and miRNA-21 in serum of patients with prostate cancer was detected by RT-PCR.Results:After treatment, the prostate volume and serum prostate specific antigen (PSA) expression were significantly reduced(3 months after treatment compared with before treatment, prostate volume: t=6.22, P=0.00; PSA: t=5.14, P=0.00; 6 months after treatment compared with before treatment, prostate volume: t=8.32, P=0.00; PSA: t=4.11, P=0.00; 12 months after treatment compared with before treatment, prostate volume: t=10.14, P=0.00; PSA: t=9.25, P=0.00; 6 months after treatment compared with 3 months after treatment, prostate volume: t=3.82, P=0.00; PSA: t=6.23, P=0.00; effective rate: χ 2=6.38, P=0.00.One year after treatment compared with 3 months after treatment, prostate volume: t=4.16, P=0.00; PSA: t=5.77, P=0.00; effective rate: χ 2=5.74, P=0.00.One year after treatment compared with 6 months after treatment, the prostate volume: t=8.11, P=0.00; PSA: t=4.92, P=0.00; effective rate: χ 2=4.44, P=0.00). And with the extension of treatment time, the treatment effect was significantly enhanced.RT-PCR results showed that compared with the control group, the expression levels of miRNA-125b and miRNA-21 in the case group were significantly increased( t=6.22, 6.88, P=0.01, 0.00). After treatment with bicalutamide, the levels of miRNA-125b and miRNA-21 in serum of prostate cancer patients were significantly reduced. Conclusion:Bicalutamide can reduce the pain and increase the efficacy of prostate cancer patients; bicalutamide can reduce the expression of miRNA-125b and miRNA-21 in serum of patients with prostate cancer.
6.The efficacy of intravesical instillation of domestic BCG versus epirubicin in the prevention of recurrence of intermediate-risk or high-risk non-muscular invasive bladder cancer and predictive factors of BCG instillation: a randomized, controlled, multi-center clinical trial with 2 years’ follow-up
Hao YU ; Kaiwen LI ; Hailong HU ; Xiang LI ; Nan LIU ; Jian ZHANG ; Xudong YAO ; Xiaodong ZHANG ; Wei LI ; Liqun ZHOU ; Xiangbo KONG ; Jinjian YANG ; Youhan CAO ; Junli WEI ; Jiacun CHEN ; Zhaoyang WU ; Dongwen WANG ; Xuhui ZHANG ; Jinkai SHAO ; Qingwen LI ; Huiqing ZHANG ; Xiaolin WANG ; Shaozhong WEI ; Ye TIAN ; Tie ZHONG ; Hongshun MA ; Kun LI ; Benkang SHI ; Jin YANG ; Yuhua QIAO ; Hongxing HUANG ; Liming LI ; Zhimin WANG ; Jianhua TIAN ; Tianxin LIN ; Jian HUANG
Chinese Journal of Urology 2020;41(10):724-730
Objective:To investigate the 2 years’ efficacy of intravesical instillation of domestic BCG versus epirubicin in the prevention of recurrence of intermediate-risk or high-risk non-muscular invasive bladder cancer and predictive factors of BCG instillation.Methods:From July 2015 to June 2020, 18-75 years old patients with moderate to high-risk non muscle invasive bladder cancer (NMIBC) confirmed by pathological examination were involved. The ECOG score was 0-2. Exclusion criteria included ①immune deficiency or impairment (such as AIDS), using immunosuppressive drugs or radiotherapy, suspected allergic to BCG or epirubicin or excipients of the two drugs, fever or acute infectious diseases including active tuberculosis or receiving anti tuberculosis treatment, with severe chronic cardiovascular and cerebrovascular diseases or chronic kidney disease; ②combined with other urogenital system tumors or other organ tumors; ③combined with muscle invasive bladder urothelial carcinoma (≥T 2); ④undergoing chemotherapy, radiotherapy or immunotherapy within 4 weeks (immediate instillation after surgery not included); ⑤ pregnant or lactating women; ⑥ comfirmed or suspected bladder perforation; ⑦gross hematuria; ⑧cystitis with severe bladder irritation that may affect the evaluation; ⑨participat in other clinical trials within 3 months; ⑩alcohol or drug addiction; ?any risk factors that may increasing the risk of patients. Epirubicin 50 mg was irrigated immediately after the operation(TURBT or laser resection). The patients were randomly divided into BCG15 group, BCG19 group and epirubicin group by the ratio of 2∶2∶1, and the patients were maintained intravescical instillation for 1 year. The recurrence and adverse events of the three groups were compared. Univariate and multivariate analysis was performed to predict the risk factors of BCG irrigated therapy failure. Result:By June 15, 2020, the median follow-up duration was 22.1 months(12.1, 32.3), and there was no statistical difference between the groups ( P=0.9024). There were 274 patients enrolled in BCG19 group, 277 patients enrolled in BCG15 group and 130 patients enrolled in the epirubicin group. The drop-off rate was 16.6%(113 cases)and made no difference between groups( P=0.6222). There were no significant difference in age, gender, BMI, or ECOG score( P>0.05). During the follow-up, 116 cases was detected recurrence or progression. The recurrence rate of the three groups was 14.2% and 14.8% in BCG19 group and BCG15 group, and 27.7% in the epirubicin group. There was no difference in recurrence rate between BCG19 and BCG15 group( P=0.9464). The recurrence rate of BCG19 group was lower than that of the epirubicin group ( P=0.0017). The recurrence rate of BCG15 group was lower than that of the epirubicin group ( P=0.0020). There was no difference in the cumulative recurrence free survival rate between BCG19 and BCG15 group (95% CI0.57-1.46, P=0.7173). The cumulative recurrence free survival rate of BCG 19 group was better than that of the epirubicin group( HR=0.439, 95% CI0.26-0.74, P=0.0006), and the cumulative recurrence free survival rate of BCG15 group was better than that of the epirubicin group ( HR=0.448, 95% CI0.29-0.80, P=0.0021). The total incidence of adverse events in 19 BCG19, BCG15 and epirubicin group were 74.5%, 72.6% and 69.8% respectively. There was no difference in the incidence of adverse events between BCG19 and BCG15 group( P=0.6153). The incidence of adverse events in epirubicin group was lower than that of BCG19( P=0.0051) and BCG15( P=0.0167) groups.There was no significant difference in the incidence of serious adverse events (SAE) among the three groups ( P=0.5064). Log rank test univariate analysis and Cox risk regression model multivariate analysis showed that the history of bladder cancer recurrence( HR=6.397, 95% CI1.95-20.94, P=0.0001)was independent risk factor for BCG irrigation failure. Conclusions:The 2 years’ efficacy of intravesical instillation of domestic BCG is better than than of epirubicin with good tolerance and safety. There is no difference between BCG19 and BCG15 group. BCG doesn’t increase SAE compared with epirubicin. Recurrence status was an independent prognostic factor regarding recurrence-free survival.
7.The efficacy and safety of BCG in the prevention of postoperative recurrence of intermediate and high-risk non-muscle invasive bladder cancer: a randomized, controlled, multi-center clinical trial (mid-term report)
Hao YU ; Tianxin LIN ; Xiang LI ; Hailong HU ; Nan LIU ; Jian ZHANG ; Xudong YAO ; Zhaoyang WU ; Xiangbo KONG ; Liqun ZHOU ; Jiacun CHEN ; Wei LI ; Jinjian YANG ; Dongwen WANG ; Xiaodong ZHANG ; Youhan CAO ; Shaozhong WEI ; Ye TIAN ; Huiqing ZHANG ; Benkang SHI ; Zhanpo YANG ; Qingwen LI ; Jinkai SHAO ; Tie ZHONG ; Xiaolin WANG ; Hongxing HUANG ; Liming LI ; Jianhua TIAN ; Zhimin WANG ; Jin YANG ; Lin QI ; Jian HUANG
Chinese Journal of Urology 2019;40(7):485-491
Objective To investigate the efficacy and safety of intravesical instillation of BCG vaccine in the prevention of early recurrence of middle and high risk non-muscle invasive bladder cancer.Methods From July 2015,patients with non-muscle invasive bladder cancer aged 18-75 years with informed consent were screened and underwent transurethral resection of bladder tumor (TURBT).Immediately intravesical instillation of epirubicin 50 mg was given postoperatively.After pathology was comfirmed,patients was enrolled in group 1 (BCG15) or group 2 (BCG 19) or the control group (epirubicin 18) randomly with SAS 9.3 software.Data of follow-up and Adverse event was collected and analyzed.Results By May 31,2019,531 patients were enrolled in the study.The drop-off rate was 20.1%.167 patients (143 males and 24 females)in group 1,172 patients (141 males and 31 females)in group2 and 84(75 males and 9 females) in the control group with follow-up data were analyzed.There were no significant differences in age,gender,BMI,ECOG score,risk stratification between the three groups (P =0.8641,P =0.2906,P =0.9384,P =0.6126).The median follow-up time makes no statistical difference between the groups (P =0.9251),12.0 (6.0,22.5) months,13.0 (6.0,22.3) months,and 13.0 (7.0,22.3) months.The median recurrence time of the three groups was 4.0 (3.0,6.0) months,4.5 (3.0,9.8) months,4.5 (3.0,8.8) months.There was no statistical difference between the three groups (P =0.2852).Risk stratification in the patients got no significant difference between the three groups (P > 0.05).The 1-year recurrence-free survival rates were 80.0% in the group 1 and 88.3% in the group 2 and 73.7% in the control group.The group 2 was superior to the group 1 and the control group (P =0.0281,P =0.0031).There was no significant difference between group 1 and control group (P =0.2951).There was no significant difference in the cumulative recurrence-free survival between the experimental group 1 and the experimental group 2,(95% CI 0.80-2.43,P =0.2433).The cumulative recurrence-free survival in the group 1 and the group 2 was better than the control group (95 % CI 0.31-0.92,P =0.0266;95 % CI 0.20-0.65,P =0.0008).All the cases underwent instillation were analyzed for adverse events.The incidence of overall AE(adverse events) in group 1 was 68.5% (152/222),the incidence of grade Ⅰ-Ⅱ AE was 53.2% (118/222),the incidence of grade Ⅲ-Ⅳ AE was 15.3% (32/222).The incidence of overall AE in the group 2 was 71.8% (160/223),the incidence of grade Ⅰ-Ⅱ AE was 60.1% (134/223),and the incidence of grade Ⅲ-Ⅳ AE was 11.7% (26/223).The overall AE rate in the control group was 53.2% (59/111),of which the incidence of grade Ⅰ-Ⅱ AE was 42.4% (47/111),and the incidence of grade Ⅲ-Ⅳ AE was 10.8% (12/111).There was no difference in the incidence of overall AE between the group 1 and the group 2 (P =0.4497).The incidence of AE in the two experimental groups was higher than that in the control group (P =0.0062,P =0.0008).There was no difference in the incidence of grade Ⅲ-Ⅳ AE between the three groups (P =0.3902).Conclusions BCG(19 instillation schedule) has a better effect on preventing recurrence after 1 year of bladder surgery,which is superior to epirubicin group.The long-term efficacy of BCG in preventing recurrence and the efficacy of different schedules need to be further followed up.The lower urinary tract symptoms,which are mainly urinary frequency,are one of the causes of case fallout and should be fouced in future.Compared with epirubicin,BCG perfusion does not increase the incidence of grade Ⅲ-Ⅳ adverse reactions,and is safe to use.
8.Status quo of rescue knowledge and skills among elderly people in Guiyang City
Ting LOU ; Zhaoyang ZHONG ; Xiaoling BAI ; Zhongchen LUO ; Jin PANG
Chongqing Medicine 2018;47(7):944-946,949
Objective To investigate the status quo,influencing factors and training willingness of rescue knowledge in elderly people so as to provide the basis for formulating the elderly rescue training programs.Methods A total of 488 elderly people in three urban districts of Guiyang City were sampled by simple random and convenient sampling methods.The self-made elderly people rescue knowledge and skill status quo questionnaire was used to conduct the investigation.The general condition,training status quo and willingness were descriptively analyzed.The rescue knowledge score adopted the independent sample t test and variance analysis.Results The rescue knowledge status quo:the awareness rate of medical emergency call was the highest(66.6%),and the rest rescue awareness rate was less than 20.0 %.The training status quo and willingness:the people receiving the rescue knowledge training orly accounted for 13.3%.91.9 % and 94.1% of the elderly people considered it necessary and were willing to participate in the rescue knowledge training.The pathways obtaining the rescue knowledge:network media and families or friends introduction accounted for 37.5 % and 22.4 % respectively,followed by books and magazines,publicity column and related knowledge lectures.The influencing factors:the knowledges and skills grasping situation in the elderly people experiencing the rescue training was better than those without experience.The higher the education level,the better grasping the rescue knowledge.The grasping situation in different occupations from high to low were special technical staffs,administrative staffs,workers and farmers.Conclusion The elderly people are short of rescue knowledge,their willingness of the first aid training is strong.The government and relevant agencies should jointly perfect the first aid law and first aid training system,and formulate the rescue training program that suitable for the elderly physical and mental characteristics,increase the rescue training strength and increase the rescue ability of the elderly people.
9.A literature review on preoperative nursing of percutaneous coronary intervention for acute myocardial infarction patients
Xiaoling BAI ; Zhaoyang ZHONG ; Shiwei ZHOU ; Qinglong LIANG ; Qian FANG
Chinese Journal of Practical Nursing 2017;33(9):713-716
Objective To analyze the content, quality, the type of nursing documents and the actuality of domestic emergent percutaneous coronary intervention (PCI) in patients with acute myocardial infarction(AMI), and to provide basis and reference for the development of more objective and scientific preoperative nursing guidelines. Methods Wanfang DATA, China National Knowledge Infrastructure (CNKI), CqVIPwere retrieved by computer, and supplemented by manual searches.Then the articles which in accordance with inclusion criteria were analyzed by descriptive analysis. Results A total of 114 literatures from 2004 to 2016 were involved, most of them were presented from 2010 to 2014, all of them were published in nearly 70 kinds of journals. There were merely 9 kinds of nursing journals, 26 literatures in all. Literatures came from different levels of hospital, including 47 provincial hospitals, 50 municipal hospitals, 17county hospitals. Only 3 of them had fund support. According to the types of research, there were 61 quasi- experimental studies, 35 retrospective analysis and 18 experience summarization. The most of literatures about perioperative nursing and whole nursing were 39. Nursing experience literatures were 27, nursing cooperation or nursing intervention or nursing study were 15, others were 12. Only preoperative nursing were 4. All of this content were much the same, with minor innovativeness. Conclusions Researches of domestic nursing scholars on intraoperative and postoperative nursing of emergent PCI are more than preoperative nursing, and the attention to preoperative nursing is not high, we should strengthen the importance to preoperative nursing and increase the study of nursing before the operation in order to strive for the precious time when rescuing patients with acute myocardial infarction.
10.Clinicopathological features and prognostic significance of human epidermal growth factor receptor 2 in gastric carcinoma
Mengying KOU ; Zhaoyang WANG ; Richang DU ; Tao ZHANG ; Min FENG ; Lingli DENG ; Xiangguo ZHANG ; Chunlan ZHONG ; Lei CUI ; Jiaocheng WANG
Cancer Research and Clinic 2016;28(7):441-446
Objective To explore the clinicopathological features and prognostic significance of human epidermal growth factor receptor 2 (HER2) in gastric carcinoma. Methods Pathological data of 127 patients with gastric carcinoma were retrospectively analyzed. HER2 expressions of all patients were detected by immunohistochemistry (IHC). 119 (93.7 %) patients were undergone R0 dissection and 123 (96.9 %) cases received D2 lymph nodes dissection. 51 (40.2 %) patients received adjuvant chemotherapy. The proportional differences of clinicopathological features for patients between HER2-positive and HER2-negative were compared, including the patients' survival. Results HER2 overexpression rate was 8.7 % (11/127) in gastric carcinoma patients. For the patients with HER2-positive and HER-negative, the lymph node metastasis rates were 100.0 % (11/11) and 81.9 % (95/116), respectively (P= 0.041). The 3-year overall survival (OS) rates for HER2-positive and HER2- negative patients with gastric carcinoma were 32.7 % and 42.9 % (P=0.413), and the 5-year progression free survival (PFS) rates were 27.3 % and 42.2 % (P = 0.354), respectively. Among patients with HER2-negative, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 55.3%, compared with 35.4%for patients with surgery alone (P=0.015), and the 3-year PFS rates were 53.3 % and 35.3 % (P= 0.038), respectively. Among patients with HER2-positive patients, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 0, compared with 75.0%for patients with surgery alone (P=0.002), and the 3-year PFS rates were 0 and 60.0% (P=0.004). Conclusions HER2 is expressed in gastric carcinoma tissue, related to lymph node metastasis. HER2 status are not correlated with the prognosis for gastric carcinoma patients, however, it is likely to be a predictive marker for adjuvant treatment after surgery for patients with gastric carcinoma.

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