1.Summary of optimal evidences for early fluid resuscitation management in patients with acute pancreatitis
Li LI ; Liqun ZHU ; Wenhua ZHANG ; Yingfeng ZHOU ; Lei BAO ; Guofu SUN ; Yuanyuan MI ; Liping YANG ; Leiyuan ZHONG ; Wei ZHANG
Chongqing Medicine 2024;53(1):114-120
Objective To systematically retrieve,evaluate and integrate the best evidences on the early fluid resuscitation management in the patients with acute pancreatitis(AP)at home and abroad to provide ref-erence for clinical decision.Methods The related evidences on the early fluid resuscitation management in the AP patients were retrieved by computer from the databases of BMJ Best Practice,Up To Date,JBI,National Institute for Health and Care Excellence,Registered Nurses Association of Ontario,Guideline International Network,Scottish Intercollegiate Guidelines Network,International Association of Pancreatology,American Pancreatic Association,American College of Gastroenterology,Yimaitong,Cochrane Library,PubMed,Em-bass,CINAHL,The Web of Science,CNKI,Wanfang databases.The retrieval time limit was from the data-base establishment to March 20,2022.The literatures types included thematic evidence summarization,guide-lines,evidence summaries,systematic reviews and expert consensus.The researchers conducted the literature quality evaluation.The literatures meeting the standard conducted the evidence extraction.Results A total of 13 arti-cles were included,including 3 special subject evidence summary,4 guidelines,2 evidence summary,2 systematic evalu-ation and 2 expert consensus.A total of 16 pieces of best evidence were integrated,involving 4 aspects of organization management,evaluation and monitoring,fluid infusion strategy and health education.Conclusion It is recommended to use the target-oriented therapy for early fluid resuscitation management,and perform the fluid resuscitation immediate-ly after diagnosis,according to the patient's underlying disease,disease changes and monitoring indicators,implement precise early fluid resuscitation in order to reverse pancreatic microcirculation disorder,increase tissue perfusion and improve the patient's prognosis.
2.Methodology for Developing Patient Guideline (3):Reporting Frameworks and Presentation
Lijiao YAN ; Ning LIANG ; Haili ZHANG ; Nannan SHI ; Ziyu TIAN ; Ruixiang WANG ; Xiaojia NI ; Yufang HAO ; Wei CHEN ; Yingfeng ZHOU ; Dan YANG ; Shuyu YANG ; Yujing ZHANG ; Ziteng HU ; Jianping LIU
Journal of Traditional Chinese Medicine 2024;65(22):2304-2309
Standardized reporting is a crucial factor affecting the use of patient guidelines (PGs), particularly in the reporting and presentation of recommendations. This paper introduced the current status of PG reporting, including the research on PG content and presentation formats, and provided comprehensive recommendations for PG reporting from aspects such as overall framework, recommendations, presentation format, and readability. First, the presentation of PG recommendations should include clearly defined clinical questions, recommendations and their rationale, and guidance on how patients should implement the interventions; for specific content in the PG, such as level of evidence, level of recommendation, it is recommended to explain in text the reasons for giving different levels of recommendation, i.e., to present the logic behind giving the level of recommendation to the patient; additional information needed in the recommendation framework should be supplemented by tracing references or authoritative textbooks and literature that support the recommendations. Subsequently, the PG text should be written based on the Reporting Checklist for Public Versions of Guidelines (RIGHT-PVG) reporting framework. Finally, to enhance readability and comprehension, it is recommended to refer to the Patient Education Materials Assessment Tool (PEMAT) for translating PG content. To enhance the readability of PGs, it is suggested to present the PG content in a persona-lized and layered manner.
3.Preparation of mouse monoclonal antibodies against the ectodomain of Western equine encephalitis virus E2 (E2ecto) protein.
Fuxing WU ; Yangchao DONG ; Jian ZHANG ; Pan XUE ; Ruodong YUAN ; Yang CHEN ; Hang YUAN ; Baoli LI ; Yingfeng LEI
Chinese Journal of Cellular and Molecular Immunology 2024;40(1):62-68
Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×104 were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 105, and 3D5B7 reached 107. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses
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Mice
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Encephalitis Virus, Western Equine
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Ascites
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Immunosuppressive Agents
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Antibodies, Monoclonal
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Immunoglobulin M
4.Methodology for Developing Patient Guideline(1):The Concept of Patient Guideline
Lijiao YAN ; Ning LIANG ; Ziyu TIAN ; Nannan SHI ; Sihong YANG ; Yufang HAO ; Wei CHEN ; Xiaojia NI ; Yingfeng ZHOU ; Ruixiang WANG ; Zeyu YU ; Shuyu YANG ; Yujing ZHANG ; Ziteng HU ; Jianping LIU
Journal of Traditional Chinese Medicine 2024;65(20):2086-2091
Since the concept of patient versions of guidelines (PVGs) was introduced into China, several PVGs have been published in China, but we found that there is a big difference between the concept of PVG at home and abroad, and the reason for this difference has not been reasonably explained, which has led to ambiguity and even misapplication of the PVG concept by guideline developers. By analyzing the background and purpose of PVGs, and the understanding of the PVG concept by domestic scholars, we proposed the term patient guidelines (PGs). This refers to guidelines developed under the principles of evidence-based medicine, centered on health issues that concern patients, and based on the best available evidence, intended for patient use. Except for the general attribute of providing information or education, which is typical of common health education materials, PGs also provide recommendations and assist in decision-making, so PGs include both the patient versions of guidelines (PVG) as defined by the Guidelines International Network (GIN) and "patient-directed guidelines", i.e. clinical practice guidelines resulting from the adaptation or reformulation of recommendations through clinical practice guidelines.
5.Methodology for Developing Patient Guideline (2):Process and Methodology
Lijiao YAN ; Ning LIANG ; Nannan SHI ; Sihong YANG ; Ziyu TIAN ; Dan YANG ; Xiaojia NI ; Yufang HAO ; Wei CHEN ; Ruixiang WANG ; Yingfeng ZHOU ; Shibing LIANG ; Shuyu YANG ; Yujing ZHANG ; Ziteng HU ; Jianping LIU
Journal of Traditional Chinese Medicine 2024;65(21):2194-2198
At present, the process and methodology of patient guidelines (PGs) development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers, we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus, so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage, the construction stage, and the completion stage in general, but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network, i.e., team formation, screening of recommendations, guideline drafing, user testing and feedback, approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process (forming a team, determining the scope of the PG, searching, evaluating and integrating evidence, forming recommendations, writing the guideline, and expert review). Based on the different models, we sort out the process and methods of PG development and introduce the specific methods of PG development, including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines, with a view to providing reference for guideline developers and related researchers.
6.GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism
Hong GU ; Jia WANG ; Wenwen ZHANG ; Xiao YANG ; Xiyue HUANG ; Yingfeng ZHANG ; Yanhua MAO ; Changjiang LI ; Qiuhong CHEN ; Jinglin HUANG ; Qi LIU ; Ling WEI ; Congcong SUN
Journal of Army Medical University 2024;46(18):2110-2120
Objective To investigate the effect of glycine N-methyl transferase (GNMT)on intrauterine adhesion (IUA)fibrosis and its related mechanism.Methods In vivo experiment:A total of 36 healthy female SD rats (SPF grade,6~8 weeks old and weighing from 180~220 g)were subjected in this study.IUA model of SD rats and IUA model of GNMT overexpressed rats were established.RT-qPCR and immunofluorescence assay were applied to detect GNMT expression level in normal uterus and model group.RT-qPCR and Western blotting were used to detect the mRNA and protein levels of fibrosis-related molecules and the activation of TGF-β1/Smad3 signaling pathway in each group.The number of endometrial glands in each group was observed by HE staining.Masson staining was used to analyze the severity of endometrial fibrosis in each group.In vitro experiment:transformed human endometrial stromal cells (THESCs)fibrotic phenotype model was constructed using TGF-β1,and THESCs stably transfected with GNMT overexpression lentvirus were treated with TGF-β1.RT-qPCR and Western blotting were used to detect the mRNA and protein expression of fibrosis-related molecules.The expression of TGF-β1/Smad3 signaling pathway was detected by Western blotting.TGF-β1/Smad3 signaling pathway was activated by TGF-β1/Smad signaling pathway activator (SRI-011381),and the expression of TGF-β1/Smad3 signaling pathway and key molecular proteins of fibrosis phenotype was measured with Western blotting.Results In vivo experiment,the mRNA and protein expression levels of GNMT were significantly decreased in the IUA rats than the control rats (P<0.05).Overexpression of GNMT decreased the mRNA and protein levels of fibrosis related molecules,Collagen Ⅰ,Collagen Ⅲ and FN in the IUA rats (P<0.05),and decreased the phosphorylation levels of TGF-β1 and its downstream Smad3 protein (P<0.05).HE and Masson staining showed that overexpression of GNMT could increase the number of endometrial glands and reduce the severity of fibrosis in the IUA rats (P<0.05).In vitro experiments:overexpression of GNMT decreased the mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ and FN associated with fibrotic phenotype of THESCs (P<0.05),and reduced the phosphorylation level of Smad3 protein,downstream of TGF-β1 (P<0.05).After activation of TGF-β1/Smad3 signaling pathway,the protein levels of TGF-β1/Smad3 signaling pathway and downstream fibrosis phenotype molecules,Collagen Ⅲ and FN,were significantly decreased in the LV-GNMT+SRI-011381 group.Conclusion Overexpression of GNMT can inhibit endometrial fibrosis by regulating TGF-β1/Smad3 signaling pathway,thus achieving therapeutic effect on IUA.
7.Dahuang Mudantang Alleviates Intestinal Injury in Rat Model of Acute Pancreatitis by Regulating HMGB1/RAGE/NF-κB Signaling Pathway
Linlin WEN ; Yongfeng WANG ; Min BAI ; Zhandong WANG ; Bing SONG ; Yanying ZHANG ; Xinhong LIU ; Yingfeng SUN ; Dan YANG ; Qiong WANG ; Hongzhang ZHAO ; Runze YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(19):1-8
ObjectiveTo explore the mechanism of Dahuang Mudantang in alleviating the intestinal injury in the rat model of acute pancreatitis via the high-mobility group box 1 (HMGB1)/receptor for advanced glycation endproduct (RAGE)/nuclear factor-κB (NF-κB) signaling pathway. MethodOne hundred and twenty SPF-grade Wistar rats received retrograde injection of 5% sodium taurocholate into the biliopancreatic duct for the modeling of intestinal injury in acute pancreatitis. The rats were randomized into blank, model, low-, medium-, and high-dose (3.5, 7, 14 g·kg-1, administrated by gavage) Dahuang Mudantang, and octreotide (1×10-5 g·kg-1, subcutaneous injection) groups (n=20). The rats in blank and model groups received equal volume of distilled water by gavage. Drugs were administered 1 h before and every 12 h after modeling, and samples were collected 24 h after modeling. The general status of the rats was observed. The biochemical methods were employed to measure the levels of amylase (AMS) and C-reactive protein (CRP) in the serum. The enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in the colon tissue. The morphological changes of pancreatic and colon tissues were observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to measure the expression levels of HMGB1, RAGE, inhibitor of NF-κB kinase (IKK), and NF-κB suppressor protein α(IκBα)in the colon tissue. ResultThe rats in the model group showed poor general survival, writhing response, reduced frequency of defecation, and dry stool. The symptoms of rats in the model group were mitigated in each treatment group, and the high-dose Dahuang Mudantang showed the most significant effect. Compared with the normal group, the model group had elevated AMS and CRP levels (P<0.05), which were lowered by Dahuang Mudantang (P<0.05), especially that at the high dose (P<0.05). Compared with the normal group, the modeling elevated that levels of TNF-α, IL-1β, and IL-6 (P<0.05). Such elevations were lowered by Dahuang Mudantang (P<0.05), and the high-dose group and the octreotide group showed better performance (P<0.05). The modeling caused necrotic, congested, and destructed pancreatic and colonic tissues, which were ameliorated by the drugs, especially high-dose Dahuang Mudantang. Compared with the normal group, the modeling up-regulated the mRNA levels of HMGB1, RAGE, IKK, IκBα, and NF-κB (P<0.05). Compared with the model group, Dahuang Mudantang and octreotide down-regulated the mRNA levels of HMGB1, RAGE, IKK, IκBα, and NF-κB (P<0.05), and the high-dose Dahuang Mudantang demonstrated the best performance (P<0.05). Western blot results showed a trend consistent with the results of Real-time PCR. ConclusionDahuang Mudantang can improved the general status, reduce inflammation, and alleviate histopathological changes in the pancreatic and colon tissues in the rat model of acute pancreatitis by inhibiting the HMGB1/RAGE/NF-κB signaling pathway.
8.Survey on awareness of occupational health knowledge and concept among front-line workers in the automotive manufacturing industry
Xinyang YU ; Min YANG ; Huiqing CHEN ; Junle WU ; Hao LUO ; Yingfeng GE ; Yuduan HAN ; Jinxin ZHANG
China Occupational Medicine 2023;50(4):410-414
Objective To investigate the awareness of occupational health knowledge and concept of front-line workers in the automobile manufacturing industry. Methods A total of 1 188 front-line workers from 47 automobile manufacturing enterprises in Guangdong Province were selected as the research subjects using the judgment sampling method. The awareness of occupational health knowledge and concept of the workers was investigated using the Personal Questionnaire for National Survey of Occupational Health Literacy for Key Populations. Results The median (25th and 75th percentiles) awareness of occupational health knowledge and concept among the research subjects was 56.00% (44.00%, 64.00%). The awareness was higher among male workers than female workers (P<0.05). The awareness was higher among married workers than workers with other marital status (P<0.05). The awareness of workers with an education level of middle school or below was lower than those in the senior high school and junior college group and bachelor's degree or above group (all P<0.05). The awareness of workers in the group with personal monthly income <5 000 was lower than that in the other two income groups with personal monthly income ≥5 000 (all P<0.05). The awareness of workers in medium and large enterprises was higher than that of workers in small and micro enterprises (all P<0.05). The awareness of workers in foreign-funded enterprises was higher than that in private enterprises and other enterprises (all P<0.05). The awareness of workers with job tenure >5-10 and >10 years was higher than that in workers with job tenure of 1-≤5 years (P<0.05). The awareness for the 25 items of occupational health knowledge and concept among the research subjects ranged from 6.14% to 96.72%, with an average awareness of 54.46% for the 25 items. Conclusion The awareness of occupational health knowledge and concept of front-line workers in automobile manufacturing industry is not high in general, and it is influenced by gender, marriage status, education level, personal monthly income, enterprise nature and scale, and job tenure.
9.On the cultivation of medical students' humanistic quality from the perspective of virtual simulation teaching
Baojie WU ; Xiaoqin YANG ; Yijia LAI ; Yingfeng LIN ; Xueling ZHANG ; Jingshu DUAN
Chinese Journal of Medical Education Research 2022;21(8):1008-1011
Based on the characteristics of medical specialty, this research group has developed the virtual simulation experiment teaching project of Chinese revolutionary morality (red medicine spirit). This project uses 3D simulation technology and OpenGL technology to build a virtual experimental learning environment based on WebGL, learn general Fu Lianzhang's glorious deeds and immersive experience of the inheritance and development of the spirit of red medicine by contemporary "heroes in harms way" in the fight against the COVID-19 epidemic in the form of 3D animation interaction. Through research, it is found that virtual simulation teaching is conducive to promoting students' understanding and mastery of medical humanities and improving students' satisfaction with teaching, with good popularization value.
10.Analysis of differences and influencing factors of liver injury associated with different strains of 2019-nCoV infection
Guodong YU ; Jiangshan LIAN ; Chanyuan YE ; Feng DING ; Yingfeng LU ; Shaorui HAO ; Jiong YU ; Yida YANG
Chinese Journal of Hepatology 2022;30(5):520-526
Objective:To analyze whether there are differences and related influencing factors in liver injury associated with different strains of 2019-nCoV/SARS-CoV-2 infection.Methods:Data of epidemiology, clinical symptoms, laboratory tests, and treatment outcomes of patients with COVID-19 infection confirmed with Alpha and Delta virus strain in Zhejiang Province were retrospectively collected. Statistical analysis was performed using independent samples t-test or Mann-Whitney U test, χ2 test or Fisher's exact test, and logistic regression analysis. Results:A total of 788 and 381 cases with Alpha and Delta virus strain were included. Vaccination ratio was 0% in Alpha and 85.30% in Delta group ( P<0.001), The proportion of patients with fever (80.71% vs. 40.94%, P<0.001) was significantly higher in Alpha than Delta strain group. The proportion of critical ill patients was significantly higher in Delta group (9.90% vs. 1.57%, respectively, P<0.001). The virus negative conversion time was significantly longer in Delta than Alpha group (22 d vs. 11 d, P<0.001), but the incidence of liver injury was significantly higher in Alpha than Delta group (20.05% vs. 13.91%, P=0.011). Univariate analysis showed that Alpha virus strain infection, male sex, body mass index, chronic liver disease, fever, diarrhea, shortness of breath, severe/critical illness, elevated creatine kinase (CK), elevated international normalized ratio (INR) and an elevated neutrophil/lymphocyte ratio was significantly associated with an increased risk of liver injury occurrence, and in patients with pharyngeal pain the risk of liver injury occurrence was significantly reduced. Multivariate analysis showed that shortness of breath [ OR, 2.667 ( CI: 1.389-5.122); P=0.003], increased CK [ OR, 2.544 ( CI: 1.414-4.576); P=0.002] and increased INR [OR, 1.721] ( CI: 1.074-2.758); P=0.024] was significantly associated with an increased risk of liver injury occurrence, and in patients with pharyngeal pain the risk of liver injury occurrence was significantly reduced [ OR, 0.424 ( CI: 0.254-0.709); P=0.001]. Conclusion:Although the virulence of the Delta is stronger than Alpha strain, most patients infected with Delta strain vaccinated against COVID-19 in Zhejiang province had milder clinical symptoms and a lower incidence and degree of liver injury. Notably, the infection risk even remains after vaccination; however, symptoms and the incidence of severe and critical illness can be significantly reduced.

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