1.Construction and optimization of a novel plasma membrane model for membrane affinity assay based on polydiacetylene vesicles
Chai-chai NIE ; Rui-ting DONG ; Yu-tong WU ; Jing-bo WU ; Sheng ZHANG ; Feng ZHENG ; Ya DING
Acta Pharmaceutica Sinica 2023;58(8):2503-2511
Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.
2.Observation of IL-17 levels of perioperative period in congenital heart disease patients with pulmonary hypertension
qing Xiao FAN ; ting Rui WANG ; qing Xiao CHAI ; long De WANG ; qiang Ke HE
Tianjin Medical Journal 2017;45(10):1036-1039
Objective To discuss the changes and significance of interleukin-17 (IL-17) in perioperative period of congenital heart disease patients with pulmonary hypertension. Methods A total of forty patients with congenital heart disease underwent cardiopulmonary bypass (CPB) were included in this study. According to the pulmonary artery systolic pressure (PASP), patients were divided into non-pulmonary hypertension group (group Ⅰ, PASP < 30 mmHg) and pulmonary hypertension group (groupⅡ, PASP≥30 mmHg). Blood samples were taken before anesthesia (T1), start CPB (T2), 30 min after CPB (T3), 6 h (T4), 24 h (T5) and 7 d (T6) after operation. The concentration of IL-17 was detected by ELISA. Arterial oxygen partial pressure [p(O2)] and arterial carbon dioxide partial pressure [p(CO2)] during the first five time points were recorded. Oxygen index (OI) and alveolar arterial oxygen tension difference (AaDO2) were calculated. Results The plasma IL-17 levels in perioperative period were significantly higher in group Ⅱ than those of group Ⅰ (P < 0.05). The highest concentration of IL-17 emerged at T3, then decreased gradually in both groups. At this time point, the OI decreased, and AaDO2 increased significantly in both groups. Compared with groupⅠ, the OI decreased, while AaDO2 increased at T5 in groupⅡ(P<0.05). Conclusion The high level of IL-17 promotes the formation of pulmonary hypertension in congenital heart disease and leads to the lung injury during CPB, which can be used as a clinical monitoring indicator of evaluating severity.
3.Radiosensitivity of lncrna linc00909 targeting mir-548-3p on colorectal cancer cells
Ge HOU ; Chenhu XIAO ; Xiaojuan CHEN ; Yangyang HUANG ; Cheng WANG ; Ting CHAI ; Rui SONG ; Jinjin YUAN ; Zongwen LIU
Chinese Journal of Radiation Oncology 2020;29(12):1110-1117
Objective:To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods:The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results:The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated ( P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated ( P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced ( P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced ( P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p ( P<0.05). Conclusion:Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.
4.LncRNA CCAT1 enhances radiosensitivity of human pancreatic cancer cells PANC-1 by targeting miR-130b-3p
Rui SONG ; Jinjin YUAN ; Ge HOU ; Jun YANG ; Xiaojuan CHEN ; Ting CHAI ; Cheng WANG ; Zongwen LIU
Chinese Journal of Radiological Medicine and Protection 2019;39(4):247-254
Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1.Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays.After silencing CCAT1 and/or inhibiting miR-130b-3p expression,cell apoptosis rate,Caspase 3 activity and cell survival were detected by flow cytometry,Caspase 3 activity detection kit and colony formation assay,respectively.Cell survival curve was stimulated by the multi-target single-hit model.Based on the starBase v2.0 online analysis,the luciferase reporter gene assay,RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p.Results CCAT1 expression was up-regulated (t=6.322-8.555,P<0.05),but miR-130b-3p expression was down-regulated (t =3.950-18.795,P< 0.05) in the radiation-resistant pancreatic cancer tissues,pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells.When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy,cell survival fraction decreased (t=2.929,5.047,5.234,5.125,P<0.05),apoptosis rate and Caspase 3 activity increased (t=6.953,6.836,P<0.05).CCAT1 could selectively regulate miR-130b-3p expression.Inhibition of miR-130b-3p expression could enhance PANC-l cell survival (t =4.564,6.736,8.656,P<0.05),but reduced apoptosis rate (t=5.234,P<0.05) and Caspase 3 activity (t=10.440,P<0.05).Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells.
5.Effects of probiotics on intestinal flora, intestinal function and T lymphocyte level in patients with cervical cancer after radiotherapy
Ting CHAI ; Songjie ZHANG ; Rui SONG ; Yanli DUANMU ; Jinjin YUAN ; Ge HOU ; Alan CHU ; Yangyang HUANG ; Shijia LIU ; Zongwen LIU
Chinese Journal of Primary Medicine and Pharmacy 2023;30(7):1012-1017
Objective:To investigate the effects of probiotics on intestinal flora, intestinal function, and T lymphocyte level in patients with cervical cancer after radiotherapy.Methods:A total of 92 patients with cervical cancer who underwent pelvic radiotherapy in The Second Affiliated Hospital of Zhengzhou University from September 2020 to February 2022 were included in this study. They were randomly divided into control and experimental groups ( n = 46/group). The patients in the experimental group took probiotics during radiotherapy, while the patients in the control group did not take probiotics during radiotherapy. The amount of intestinal flora, D-lactic acid, diamine oxidase, and T lymphocyte subset levels pre- and post-radiotherapy were compared between the two groups. Urinary lactulose (L) and mannitol (M) concentrations were determined in each group. Urinary excretion ratios of L to M were calculated. Results:After 10, 15, and 20 times of radiotherapy and after all radiotherapies, the amount of Escherichia coli and Enterococcus in the experimental group was significantly lower than that in the control group ( F = 128.60, 224.99, all P < 0.05). The amount of Bifidobacteria and Lactobacilli in the experimental group was significantly higher than that in the control group ( F = 2 065.46, 948.23, both P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma D-lactic acid level in the experimental group was (9.34 ± 1.63) μg/L, (9.15 ± 1.36) μg/L, (8.68 ± 1.06) μg/L, and (8.05 ± 0.82) μg/L, respectively. After 10, 15, and 20 times of radiotherapy and after all radiotherapies, plasma diamine oxidase level in the experimental group was (86.34 ± 20.25) μg/L, (84.28 ± 17.45) μg/L, (80.40 ± 13.35) μg/L, and (76.85 ± 10.87) μg/L, respectively, and urinary excretion ratio of L to M in the experimental group was (1.84 ± 0.16), (1.55 ± 0.12), (1.26 ± 0.09), (0.98 ± 0.06), respectively, all of which were significantly lower than those in the control group ( F = 121.60, 31.73, 417.84, all P < 0.05). After 10, 15, and 20 times of radiotherapy and after all radiotherapies, CD4 + level in the experimental group was (39.80 ± 4.90)%, (40.92 ± 5.30)%, (42.52 ± 6.14)%, (43.83 ± 6.55)%, respectively, CD4 +/CD8 + was (1.52 ± 0.25), (1.63 ± 0.22), (1.71 ± 0.39), (1.83 ± 0.22), respectively, all of which were significantly higher than those in the control group ( F = 58.69, 31.07, all P < 0.05). Conclusion:Probiotics can improve the status of intestinal flora and intestinal barrier function in patients with cervical cancer after radiotherapy, and simultaneously improve the cellular immune function of patients.
6.Effect of Shaoyaotang on Expressions of CD14, FADD and Caspase-8 in Colonic Tissues of Rats with Large Intestinal Damp-heat Syndrome of Ulcerative Colitis
Si-qi CAO ; Feng-yi WANG ; Sheng-nan TANG ; Dang-sheng ZHAO ; Yang-yang LI ; Zhi-jie LIU ; Rui-ting CHAI
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(5):1-7
Objective:To observe the effect of Shaoyaotang on the contents of cell adhesion molecule-1 (ICAM-1) and transforming growth factor-
7.Application of SifaInDel 45plex System in the Han and Mongolian Populations.
Guang-Yuan YANG ; Chun-Yan YUAN ; Rui-Yang TAO ; Ruo-Cheng XIA ; Ya-Li WANG ; Xin-Yu DONG ; Si-Yu CHAI ; Li-Ming WU ; Zhi-Wei JIANG ; Ting-Ting JIANG ; Kai-Qin CHEN ; Cheng-Tao LI ; Li-Qin CHEN
Journal of Forensic Medicine 2022;38(6):733-738
OBJECTIVES:
To investigate the genetic polymorphism of InDel loci in SifalnDel 45plex system in the Han population in Jiangsu Province and the Mongolian population in Inner Mongolia, and to evaluate the effectiveness of the system in forensic medicine.
METHODS:
SifaInDel 45plex system was used for genotyping in blood samples of 398 unrelated individuals from the above two populations, and allele frequencies and population genetic parameters of the two populations were calculated respectively. Eight intercontinental populations in the gnomAD database were used as reference populations. The genetic distances between the two studied populations and eight reference populations were calculated based on the allele frequencies of 27 autosomal-InDels (A-InDels). The phylogenetic trees and multidimensional scaling (MDS) analysis diagrams were constructed accordingly.
RESULTS:
Among two studied populations, the 27 A-InDels and 16 X-InDels showed no linkage disequilibrium between each other and the allele frequency distributions were in Hardy-Weinberg equilibrium. The CDP of the 27 A-InDels in two studied populations were all higher than 0.999 999 999 9, and the CPEtrio were all less than 0.999 9. The CDP of the 16 X-InDels in Han in Jiangsu and Mongolian in Inner Mongolia female and male samples were 0.999 997 962, 0.999 998 389, and 0.999 818 940, 0.999 856 063, respectively. The CMECtrio were all less than 0.999 9. The results of population genetics showed that the Jiangsu Han nationality, Inner Mongolia Mongolian nationality and East Asian population clustered into one branch, showing closer genetic relationship. The other 7 intercontinental populations clustered into another group. And the above 3 populations displayed distant genetic relationships with the other 7 intercontinental populations.
CONCLUSIONS
The InDels in the SifaInDel 45plex system have good genetic polymorphism in the two studied populations, which can be used for forensic individual identification or as an effective complement for paternity identification, and to distinguish different intercontinental populations.
Humans
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Phylogeny
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Gene Frequency
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Polymorphism, Genetic
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Genetics, Population
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Asian People/genetics*
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China
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INDEL Mutation
8.Expert consensus on late stage of critical care management.
Bo TANG ; Wen Jin CHEN ; Li Dan JIANG ; Shi Hong ZHU ; Bin SONG ; Yan Gong CHAO ; Tian Jiao SONG ; Wei HE ; Yang LIU ; Hong Min ZHANG ; Wen Zhao CHAI ; Man hong YIN ; Ran ZHU ; Li Xia LIU ; Jun WU ; Xin DING ; Xiu Ling SHANG ; Jun DUAN ; Qiang Hong XU ; Heng ZHANG ; Xiao Meng WANG ; Qi Bing HUANG ; Rui Chen GONG ; Zun Zhu LI ; Mei Shan LU ; Xiao Ting WANG
Chinese Journal of Internal Medicine 2023;62(5):480-493
We wished to establish an expert consensus on late stage of critical care (CC) management. The panel comprised 13 experts in CC medicine. Each statement was assessed based on the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) principle. Then, the Delphi method was adopted by 17 experts to reassess the following 28 statements. (1) ESCAPE has evolved from a strategy of delirium management to a strategy of late stage of CC management. (2) The new version of ESCAPE is a strategy for optimizing treatment and comprehensive care of critically ill patients (CIPs) after the rescue period, including early mobilization, early rehabilitation, nutritional support, sleep management, mental assessment, cognitive-function training, emotional support, and optimizing sedation and analgesia. (3) Disease assessment to determine the starting point of early mobilization, early rehabilitation, and early enteral nutrition. (4) Early mobilization has synergistic effects upon the recovery of organ function. (5) Early functional exercise and rehabilitation are important means to promote CIP recovery, and gives them a sense of future prospects. (6) Timely start of enteral nutrition is conducive to early mobilization and early rehabilitation. (7) The spontaneous breathing test should be started as soon as possible, and a weaning plan should be selected step-by-step. (8) The waking process of CIPs should be realized in a planned and purposeful way. (9) Establishment of a sleep-wake rhythm is the key to sleep management in post-CC management. (10) The spontaneous awakening trial, spontaneous breathing trial, and sleep management should be carried out together. (11) The depth of sedation should be adjusted dynamically in the late stage of CC period. (12) Standardized sedation assessment is the premise of rational sedation. (13) Appropriate sedative drugs should be selected according to the objectives of sedation and drug characteristics. (14) A goal-directed minimization strategy for sedation should be implemented. (15) The principle of analgesia must be mastered first. (16) Subjective assessment is preferred for analgesia assessment. (17) Opioid-based analgesic strategies should be selected step-by-step according to the characteristics of different drugs. (18) There must be rational use of non-opioid analgesics and non-drug-based analgesic measures. (19) Pay attention to evaluation of the psychological status of CIPs. (20) Cognitive function in CIPs cannot be ignored. (21) Delirium management should be based on non-drug-based measures and rational use of drugs. (22) Reset treatment can be considered for severe delirium. (23) Psychological assessment should be conducted as early as possible to screen-out high-risk groups with post-traumatic stress disorder. (24) Emotional support, flexible visiting, and environment management are important components of humanistic management in the intensive care unit (ICU). (25) Emotional support from medical teams and families should be promoted through"ICU diaries"and other forms. (26) Environmental management should be carried out by enriching environmental content, limiting environmental interference, and optimizing the environmental atmosphere. (27) Reasonable promotion of flexible visitation should be done on the basis of prevention of nosocomial infection. (28) ESCAPE is an excellent project for late stage of CC management.
Humans
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Consensus
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Critical Care/methods*
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Intensive Care Units
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Pain/drug therapy*
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Analgesics/therapeutic use*
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Delirium/therapy*
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Critical Illness