1.Generation of MCM2 gene inducible knockout cervical cancer HeLa cells and its effect on DNA replication
Ping LI ; Tuo TANG ; Aixue ZHENG ; Luping ZHANG ; Tao WANG ; Xian HONG ; Zhihui DENG
The Journal of Practical Medicine 2024;40(2):133-139
Objective To generate minichromosome maintenance protein 2(MCM2)gene knockout cervi-cal cancer HeLa cell lines using inducible CRISPR/Cas9 technology,and to explore the effect of MCM2 on DNA replication and replication stress.Methods The inducible CRISPR/Cas9 system,TLCV2,was used to construct MCM2 knockout HeLa cell lines.And the cell lines were divided into control group(Control),knockout group 1(KO1),and knockout group 2(KO2).Western blot,Edu incorporation experiment,real-time quantitative PCR(qPCR),immunofluorescence and MTT assay were used to analyze the effects of MCM2 knockout on DNA replica-tion and replication stress induced by hydroxyurea.Results The CRISPR/Cas9 system successfully knocked out the MCM2 gene after induction,and MCM2 knockout affected the stability of MCM2-7 complex.Compared with the control cells,MCM2 knockout cells had a dramatic decrease in the capacity of DNA replication,and the mRNA levels of Cyclin A1,Cyclin E1 and CDK4.Under DNA replication stress,MCM2 knockout cells decreased cell viability,DNA damage repair capacity,and increased genomic instability compared with control cells.Conclusion Knockout of MCM2 gene reduces the DNA replication capacity of HeLa cells under normal conditions and cell viability under replication stress.This study successfully generates MCM2 gene inducible knockout HeLa cell lines,laying the foundation for further research on the role and biological function of MCM2 gene in the occurrence and progression of cervical cancer.
2.Comparison of liver and adrenal transplantation models of neuroblastoma
Hongxia CHEN ; Zhigang TAN ; Huiran LIN ; Luping FENG ; Chuya ZHENG ; Wenfeng LIAO ; Rufeng ZENG ; Jinxin LIU ; Zhenjian ZHUO
Chinese Journal of Comparative Medicine 2024;34(4):100-108
Objective A neuroblastoma(NB)liver transplantation model was established and compared with the adrenal orthotopic transplantation model to explore its characteristics.Methods 5× 105 SK-N-SH cells were implanted along the long axis of the left lobe of mouse livers with a micro-injection needle.The growth,metastasis,expression of related genes,and histopathological changes of tumors were detected after the modeling.Results The tumor formation rate in mice inoculated with tumor cells reached 100%after 21 days,and tumor growth,metastasis,related gene expression changes,and pathological characteristics were apparent.Conclusions In this study,a neuroblastoma liver transplantation model was successfully constructed via a relatively simple surgical method to provide a more suitable choice for future scientific NB experiments.
3.Application of flat-sided culture tubes during prenatal diagnosis.
Tiansheng LIU ; Hongqian HUANG ; Jiangyu SU ; Wangshang QIN ; Dongmei FEI ; Luping OUYANG ; Minpan HUANG ; Jinwu YU ; Yaqin LEI ; Shan OU ; Weijia SUN ; Qingming QIU ; Qian ZHENG
Chinese Journal of Medical Genetics 2023;40(10):1306-1311
OBJECTIVE:
To assess the value of using flat-sided culture tubes for preparing chromosomes through chorionic villi (CV) and amniotic fluid (AF) cell cultures during prenatal diagnosis.
METHODS:
From February to March 2020, 157 CV samples and 147 AF samples subjected to prenatal diagnosis at the Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region were selected as the study subjects. For each sample, one flat-sided tube and one flask culture were set up by following the standard protocols. The methods were evaluated by comparing the cell growth, experimental process, quality of chromosome preparation and costs.
RESULTS:
The success rates for the culturing of CV and AF samples by the flat-sided culture tube method were 97.45% (153/157) and 97.96% (144/147), respectively. By contrast, the success rates for the conventional flask method were 98.72% (155/157) for CV and 98.64% (145/147) for AF samples. No significant difference was found between the two methods (P > 0.05). The average harvest time required by the flat-sided culture tube method was 8.45 days for CV and 9.43 days for AF cultures, whilst the average harvest time for conventional flask method was 9.05 days and 9.54 days, respectively. The flat-sided culture tube method for CV had required significantly shorter average harvest time than the conventional method (P < 0.001). No statistical significant difference was found in the average harvest time for AF by the two methods (P > 0.05). The conventional culturing method had required three containers with two sample transfers. By contrast, the flat-sided culture tube method was carried out in one tube without any sample transfer. The average total amount of medium used was 3.91 mL for each flat-sided culture tube and 6.26 mL for each conventional flask.
CONCLUSION
The flat-sided culture tube method can provide a simple, cost-effective and error-reducing procedure for the CV and AF samples culture during prenatal diagnosis.
Child
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Female
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Pregnancy
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Humans
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China
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Prenatal Diagnosis
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Chorionic Villi Sampling
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Amniotic Fluid
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Cell Proliferation
4.Advances and Applications in Animal Models of Neuroblastoma
Zhigang TAN ; Jinxin LIU ; Chuya ZHENG ; Wenfeng LIAO ; Luping FENG ; Hongli PENG ; Xiu YAN ; Zhenjian ZHUO
Laboratory Animal and Comparative Medicine 2023;43(3):288-296
Neuroblastoma (NB) is one of the most common malignant solid tumors in children, ranks fourth in the incidence of pediatric tumors, and accounts for 15% of pediatric tumor deaths in children in China. Despite the development of new treatment options, the prognosis for high-risk patients is still poor. An animal model that can replicate the tumorigenesis of NB is an important tool for the prevention and treatment of NB. However, there are currently no animal models that can simulate all features of human NB. To provide a reference for the construction of animal models and treatment of NB, this article introduced several animal models of NB that have been extensively researched: the mouse, chick embryo chorioallantoic membrane, and zebrafish models. At the same time, it elaborated on the species, construction methods, characteristics, advantages and disadvantages, and research progress in NB.
5.Detailed resume of RNA m6A demethylases.
Dandan SHEN ; Bo WANG ; Ya GAO ; Lijuan ZHAO ; Yaping BI ; Jinge ZHANG ; Ning WANG ; Huiqin KANG ; Jingru PANG ; Ying LIU ; Luping PANG ; Zhe-Sheng CHEN ; Yi-Chao ZHENG ; Hong-Min LIU
Acta Pharmaceutica Sinica B 2022;12(5):2193-2205
N6-Methyladenosine (m6A) is the most abundant internal modification in eukaryotic mRNA, playing critical role in various bioprocesses. Like other epigenetic modifications, m6A modification can be catalyzed by the methyltransferase complex and erased dynamically to maintain cells homeostasis. Up to now, only two m6A demethylases have been reported, fat mass and obesity-associated protein (FTO) and alkylation protein AlkB homolog 5 (ALKBH5), involving in a wide range of mRNA biological progress, including mRNA shearing, export, metabolism and stability. Furthermore, they participate in many significantly biological signaling pathway, and contribute to the progress and development of cancer along with other diseases. In this review, we focus on the studies about structure, inhibitors development and biological function of FTO and ALKBH5.
6.Progress in pathophysiology and integrated traditional Chinese and Western medicine of post-intensive care syndrome
Zheng WANG ; Luping YANG ; Yidan QIN ; Yu WANG
Chinese Critical Care Medicine 2021;33(2):252-256
With the continuous improvement of treatment ability in intensive care unit (ICU), many critically ill and complex patients have survived due to the development of technology. However, most of them suffer from the psychological and physiological problems of post-intensive care syndrome (PICS). Therefore, the early identification and prevention of PICS is particularly critical. We should understand the pathophysiological mechanism of PICS, strictly implement ABCDEFG bundle management measures [including airway management (A), breath (B), reasonable analgesia and sedation treatment (C), prevention of delirium (D), early rehabilitation (E), family engagement (F), good communication (G)] during ICU hospitalization, and pay attention to reasonable nutritional support, optimizing blood glucose management, providing positive psychological intervention and support to patients through family members' encouragement and accompanying, ICU diary and other forms, following up the health status of patients transferred out/discharged from ICU, providing multi-disciplinary and professional continuing medical services, finding the entry point and timing of traditional Chinese medicine intervention, and giving full play to the advantages of integrated traditional Chinese and Western medicine treatment, which can improve the quality of life of patients. This article reviews the pathophysiological mechanism, risk factors, prevention and management measures, traditional Chinese medicine syndrome differentiation and treatment of PICS, in order to improve the early identification, diagnosis and treatment of critical patients with PICS, and improve the prognosis of patients.
7.Development of the Operating Room Professional Nurses Core Competency Evaluation Scale
Li MU ; Zheng WANG ; Luping JIANG ; Qingxin FU
Chinese Journal of Modern Nursing 2019;25(3):294-298
Objective? To develop the Operating Room Professional Nurses Core Competency Evaluation Scale so as to provide an evaluation criterion for career planning and promotion evaluation of operating room nurses. Methods? From January 2017 to March 2017, we built item pool of the scale by literature study and consulted 5 experts to form the questionnaire. From April 2017 to October 2017, we selected 443 operating room nurses attending 2017 National Operating Room Nursing Annual Meeting by convenient cluster sampling. The reliability and validity test were carried out for the original questionnaire. Items of the questionnaire were adjusted based on results to the final edition of operating room professional nurses core competency evaluation scale. Results? The final edition of the scale included 5 dimensions and 45 items involving professional development ability (12 items), professional cooperation ability (12 items), professional management ability (10 items), perioperative assessment ability (6 items) and basic professional ability (5 items). The KMO of the questionnaire was 0.975. The Cronbach's α of the scale was 0.983 and Cronbach's α of the dimensions was 0.848-0.973. Conclusions? The scale has good reliability and validity. Its content can provide a reference object for career planning of operating room nurses and an evaluation criterion for promotion evaluation of operating room nurses. The quantization of evaluation results can be a basis for operating room personnel training.
8.Analysis of the changes of PM10 concentration in surgical smoke
Li MU ; Zheng WANG ; Liwen HUANG ; Luping JIANG
Chinese Journal of Modern Nursing 2019;25(4):444-447
Objective? To investigate the changes of inhalable particulate matter (PM10) concentration in operating room and its influencing factors under different surgical methods and procedures. Methods? From June to August 2016, 23 cases of open surgery and 10 cases of laparoscopic surgery in a hospital were selected by convenience sampling method. The change of PM10 concentration during operation was monitored by P-5 digital dust meter. The measuring position was in the area of laminar air supply ceiling device (the position of the surgeon's mouth and nose is equal, and the vertical distance above the incision is 50 cm) or outside the area of the laminar air supply ceiling device (working area of itinerant nurses). The effects of different surgical methods (open surgery and endoscopic surgery) and surgical procedures (patient admission, disinfection of skin, exposure of surgical field, tissue separation, incision closure and end of operation) on the concentration of PM10 were recorded and analyzed. Results? The concentration of PM10 in 33 surgeries with different surgical procedures met the national ambient air quality standards. The average PM10 concentration of the 6 surgical procedures was (14.21±3.86)μg/m3. Comparing between open surgery and laparoscopic surgery, there were significant differences in PM10 concentration between the two groups in terms of admission, disinfection of skin, field exposure, tissue separation, incision closure and end of operation (U/t=-4.45,10.37,7.52,4.90,8.08,-4.39; P< 0.05). There were significant differences in PM10 concentration in patients whether to use high frequency electric knife or not in terms of admission,disinfection of skin, field exposure, tissue separation, incision closure and end of operation (U/t=-4.51,7.22,6.96,5.24,8.60,-4.36; P<0.05). Conclusions? The concentration of PM10 in the whole surgical process complies with national standards. The change of PM10 concentration during surgery is related to different surgical methods and the use of high frequency electric knife.
9.Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis.
Lei CHENG ; Jianjun CHEN ; Qingling FU ; Shaoheng HE ; Huabin LI ; Zheng LIU ; Guolin TAN ; Zezhang TAO ; Dehui WANG ; Weiping WEN ; Rui XU ; Yu XU ; Qintai YANG ; Chonghua ZHANG ; Gehua ZHANG ; Ruxin ZHANG ; Yuan ZHANG ; Bing ZHOU ; Dongdong ZHU ; Luquan CHEN ; Xinyan CUI ; Yuqin DENG ; Zhiqiang GUO ; Zhenxiao HUANG ; Zizhen HUANG ; Houyong LI ; Jingyun LI ; Wenting LI ; Yanqing LI ; Lin XI ; Hongfei LOU ; Meiping LU ; Yuhui OUYANG ; Wendan SHI ; Xiaoyao TAO ; Huiqin TIAN ; Chengshuo WANG ; Min WANG ; Nan WANG ; Xiangdong WANG ; Hui XIE ; Shaoqing YU ; Renwu ZHAO ; Ming ZHENG ; Han ZHOU ; Luping ZHU ; Luo ZHANG
Allergy, Asthma & Immunology Research 2018;10(4):300-353
Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult
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Asian Continental Ancestry Group*
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China
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Comorbidity
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Developed Countries
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Developing Countries
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Diagnosis*
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Epidemiologic Studies
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Epidemiology
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Global Health
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Humans
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Hypersensitivity*
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Prevalence
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Rhinitis, Allergic*
10.Content variation of four flavonoid glycosides in Hippophae rhamnoides L .leav-es before and after fermentation assayed by RP-HPLC
Ruifang DONG ; Zheng WANG ; Ling CHEN ; Zhiheng SHU ; Ting HAN ; Luping QIN
Journal of Pharmaceutical Practice 2017;35(6):526-529
Objective To compare the changes of four flavonoid glycosides in the leaves of Hippophae rhamnoides L . before and after fermentation .Methods The water extract of Hippophae rhamnoides L .leaves and its fermented tea were con-centrated and desiccated .The dry extracts were dissolved in 70% ethanol .The chromatographic separation was performed with RP-HPLC method on an Extend-C18 column (4 .6 mm × 250 mm ,5 μm) .Acetonitrile-0 .1% formic acid was selected as mobile phase at the flow rate of 1 .0 ml/min .The detection wavelength was 356 nm and the column temperature was 30 ℃ .Results The rutin content was high in the leaves of Hippophae rhamnoides L .After fermentation ,isoquercitrin content was increased , while the contents of rutin and narcissoside were reduced and isorhamnetin-3-O-glucoside stayed unchanged .There was a good linear relationship between the concentration and peak areas of the four compounds (r>0 .9997) .The average recoveries were between 96%-103% .Conclusion This established method is rapid and reliable ,which can be used for the quality control of Hippophae rhamnoides L .leaves and its fermented tea .

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