1.Expression of lncRNA SNHG8 in placenta accreta and its effect on trophoblast invasion and migration
Li'na GAO ; Xiaohui LIU ; Yufang ZHANG ; Xiaoling LIU ; Xiaochun HE ; Jing GAO ; Li ZHANG ; Jun SUN ; Xiujuan WANG ; Yan DONG
The Journal of Practical Medicine 2024;40(5):646-652
Objective To investigate the expression of lncRNA SNHG8 in placenta accrete(PA)and its effect on trophoblast invasion and migration.Methods qRT-PCR was used to detect the expression of lncRNA SNHG8 in placenta tissue of 30 cases in PA group and 30 cases in control group,and the correlation between lncRNA SNHG8 expression and prenatal ultrasound score of 30 cases in PA group was analyzed.Transwell and scratch assay were used to detect the effect of lncRNA SNHG8 interference on the invasion and migration of human chorionic trophoblast cells(HTR8/SVneo cells),and western blot was used to detect the expression of MMP-2 and MMP-9.The downstream targets of lncRNA SNHG8 were predicted by StarBase software,and the expression of lncRNA SNHG8 was detected in placental tissues of the two groups.Dual luciferase reporter assay was used to detect the targeting relationship between lncRNA SNHG8 and miR-542-3p.Results Compared with that of the control group,the expression of lncRNA SNHG8 was up-regulated in the placenta tissue of the PA group(P<0.05),and it was positively correlated with prenatal ultrasound score.Interference with lncRNA SNHG8 inhibited the invasion and migration of trophoblast cells(P<0.05);the protein expression of MMP-9 and MMP-2 also decreased signifi-cantly(P<0.05).Biological prediction indicates that miR-542-3p had a binding site with lncRNA SNHG8,and miR-542-3p expression was down-regulated in PA placental tissue(P<0.05).Dual luciferase reporter assay confirmed that lncRNA SNHG8 could target miR-542-3p.Compared with si-SNHG8+inhibitor-NC,co-transfection of si-SNHG8 and miR-542-3p inhibitor enhanced the invasion and migration ability of trophoblast cells(P<0.05).Conclusion lncRNA SNHG8 is highly expressed in PA and is related to the severity of PA.LncRNA SNHG8 promotes the invasion and migration of trophoblast by regulating the level of miR-542-3p.The study suggests that lncRNA SNHG8 plays an important role in the invasion and migration of PA trophoblast cells,which is expected to be a clinical diagnostic biomarker and therapeutic target.
2.New research direction of organ dysfunction caused by hemorrhagic shock: mechanisms of mitochondrial quality control
Zheng ZHANG ; Hongjie DUAN ; Jiake CHAI ; Xiaofang ZOU ; Shaofang HAN ; Hailiang BAI ; Yufang ZHANG ; Huiting YUN ; Ran SUN
Chinese Critical Care Medicine 2024;36(1):93-97
Hemorrhagic shock (HS) is one of the leading causes of death among young adults worldwide. Multiple organ dysfunction in HS is caused by an imbalance between tissue oxygen supply and demand, which is closely related to the poor prognosis of patient. Mitochondrial dysfunction is one of the key mechanisms contributing to multiple organ dysfunction in HS, while mitochondrial quality control regulates mitochondrial function through a series of processes, including mitochondrial biogenesis, mitochondrial dynamics, mitophagy, mitochondrial-derived vesicles, and mitochondrial protein homeostasis. Modulating mitochondrial quality control can improve organ dysfunction. This review aims to summarize the effects of mitochondrial dysfunction on organ function in HS and discuss the potential mechanisms of mitochondrial quality control, providing insights into the injury mechanisms underlying HS and guiding clinical management.
3.Establishing N-ELISA-based method for rapid detection of neutralizing antibodies against respiratory syncytial virus
Yufang SUN ; Hui ZHAO ; Huijie YANG ; Ying XIE ; Chunting BAO ; Shuyan LI ; Jiaolei WANG ; Changgui LI
Chinese Journal of Microbiology and Immunology 2024;44(2):162-169
Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.
4.Development of a working model of evidence-based nursing practice in deep vein thrombosis prophylaxis
Yu WANY ; Yufang HAO ; Yufen MA ; Yuan XU ; Ranxun AN ; Haibo DENG ; Lei WANG ; Xiaojie WANG ; Jianhua SUN ; Jia LIU ; Liyun ZHU ; Xinjuan WU
Chinese Journal of Nursing 2024;59(15):1804-1811
Objective To construct an evidence-based practice model for nurses in preventing deep vein thrombosis(DVT)and provide a scientific and targeted theoretical basis for nurses to carry out evidence-based nursing practice in DVT prevention.Methods Based on the previous evidence-based nursing practice project on DVT prevention after hip and knee arthroplasty,the research team used theoretical analysis and brainstorming to develop a draft of the work model.Expert meetings were organized to validate the content of the draft using the Delphi method,leading to the finalization of the evidence-based practice model for nurses in preventing DVT.Results The Knowledge-to-Action(KTA)framework was selected as the basic framework for constructing the evidence-based nursing practice model for preventing DVT.Theoretical Domain Framework,Theory of Planned Behavior,and Social Cognitive Theory were chosen to explore the influencing factors of nurses'behavior change in preventing DVT through evidence-based practice.The authority coefficient of the participating experts was 0.904,indicating high reliability.The final model consisted of 6 key components:knowledge generation,problem identification,localization and adaptation,knowledge application,sustained knowledge use,and conceptual framework for behavior change through evidence-based practice.Conclusion Based on theoretical analysis and clinical practice,this study developed an evidence-based practice model for nurses in preventing DVT using the expert meeting.The research methodology was scientific,and the content was reliable,providing a theoretical basis for nurses to engage in evidence-based nursing practice for DVT prevention.
5.Hemorrhagic shock-induced vascular endothelial glycocalyx damage: progress on mechanism of damage and preventive and therapeutic strategies
Yufang ZHANG ; Hongjie DUAN ; Hailiang BAI ; Ran SUN ; Huiting YUN ; Zheng ZHANG
Chinese Journal of Trauma 2023;39(9):847-856
Hemorrhagic shock, a life-threatening organ hypoperfusion caused by rapid, massive blood loss, is the leading cause of traumatic death in peacetime and wartime. The vascular endothelial glycocalyx (vEG) plays an important role in maintaining microcirculatory homeostasis. Severe ischemia and hypoxia of hemorrhagic shock can damage the vEG, leading to endothelial dysfunction and exacerbated microcirculatory and organ impairments. Therefore, early prevention and treatment of vEG damage in hemorrhagic shock can improve microcirculation dysfunction, which is of paramount importance for therapeutic efficacies and outcomes. There have been many studies on the prevention and treatment of vEG damage in hemorrhagic shock, but none is based on the management of vEG damage. The authors reviewed the progress on the mechanism and preventive and therapeutic strategies of vEG damage caused by hemorrhagic shock, hoping to provide reference for the further research of hemorrhagic shock-induced vEG damage.
6.Factors influencing the frequency of position change in adult ICU patients: a scoping review
Jiamin LI ; Xiangyu SUN ; Yufang HAO ; Xinjuan WU
Chinese Journal of Modern Nursing 2023;29(23):3199-3203
Objective:To review the influencing factors of position change frequency in adult Intensive Care Unit (ICU) patients, so as to provide reference basis for formulating position change plans for adult ICU patients.Methods:Under the guidance of the scoping review guidelines of the Joanna Briggs Institute in Australia, relevant literature was systematically searched in PubMed, ProQuest, CINAHL, China National Knowledge Infrastructure (CNKI), WanFang Data, VIP, and China Biomedical Literature Database. The search period was from database establishment to October 26, 2022. We extracted and summarized data from the included literature.Results:A total of 23 articles were included. Factors affecting the frequency of position change in adult ICU patients included patient activity ability, mobility ability, perception, moisture, nutrition, friction and shear forces, fever, skin condition, age, weight, past history, consciousness, hemodynamic status, respiration, oxygen demand, hygiene self-care ability, support surface used, patient skin response to position change frequency, and pain during position change, whether it was the final/near death stage, sleep, overall disease status, overall treatment goals, and perspective of position change.Conclusions:Nurses should fully consider relevant influencing factors when formulating a position change plan for adult ICU patients, in order to improve the prevention effect of pressure injury, promote patient recovery, and increase the cost-effectiveness of position change.
7.Mechanism of dexmedetomidine preventing sevoflurane-indued neurotoxicity to neonatal mice: the relationship with Tau phosphorylation
Mingyang SUN ; Jiaqiang ZHANG ; Ruilou ZHU ; Mengrong MIAO ; Shuang ZENG ; Yufang LENG
Chinese Journal of Anesthesiology 2022;42(3):279-283
Objective:To investigate the mechanism of dexmedetomidine preventing sevoflurane-indued neurotoxicity to neonatal mice and the relationship with Tau phosphorylation.Methods:Seventy-two SPF healthy newly born C57BL/6 wild-type mice of both sexes, aged 6 days, were divided into 4 groups ( n=18 each) using a random number table method: normal control group (C group), dexmedetomidine control group (D group), sevoflurane-induced neurotoxicity group (S group), and dexmedetomidine prevention group (SD group). Mice inhaled 2.1%-3.3% sevoflurane 2 h daily on postnatal days 6, 9 and 12, and dexmedetomidine 10 μg/kg was intraperitoneally injected at 30 min before anesthesia in group SD.Six mice were randomly selected after the end of injection, and the hippocampus tissues were removed for determination of the expression of phosphorylated Tau protein (AT8) and Tau46 protein at Tau-PS202 and Tau-PT205 sites by Western blot.The new object recognition test was performed on postnatal days 29-30 (the discrimination ratio of new objects was observed), and the Morris water maze test was performed from postnatal day 31 to 37 (the escape latency and the times of crossing the platform were observed). The hippocampi were harvested under anesthesia to detect the expression of postsynapatic density-95 by Western blot. Results:Compared with group C, the expression of AT8 was significantly up-regulated, the expression of PSD-95 was down-regulated, the number of crossing the platform and new object discrimination ratio were decreased ( P<0.05), and no significant change was found in Tau46 protein expression or escape latency in group S ( P>0.05). There was no significant difference in the indexes mentioned above between group D and group SD ( P>0.05). Compared with group S, the expression of AT8 was significantly down-regulated, the expression of postsynapatic density-95 was up-regulated, the number of crossing the platform and new object discrimination ratio were increased ( P<0.05), and no significant change was found in Tau46 protein expression and escape latency in group SD ( P>0.05). Conclusions:The mechanism of dexmedetomidine preventing sevoflurane-induced neurotoxicity to neonatal mice is related to the inhibition of Tau phosphorylation.
8. Chinese Consensus on Diagnosis and Treatment of Intestinal Behçet’s Syndrome
Hong YANG ; Wei LIU ; Weixun ZHOU ; Qingli ZHU ; Jiaming QIAN ; Yao HE ; Minhu CHEN ; Yufang WANG ; Jie LIANG ; Kaichun WU ; Qing ZHENG
Chinese Journal of Gastroenterology 2022;27(12):723-733
Behçet’s syndrome is a kind of chronic systemic vasculitis with involvement of multiple organs. Intestinal involvement of Behçet’s syndrome is presently named as intestinal Behçet’s syndrome (disease). Recently, there is considering another kind of disease type with only typical intestinal ulcers. Since it is difficult to differentiate intestinal Behçet’s syndrome from Crohn’s disease, intestinal tuberculosis, intestinal lymphoma, as well as intestinal manifestations of many other autoimmune diseases, and there is limited evidence for the therapy of intestinal Behçet’s syndrome, proposing diagnosis and treatment recommendations for intestinal Behçet’s syndrome through evidence-based judgment will be of great significance for clinical practice.
9.Reliability and validity of the Chinese version of the Treatment Burden Questionnaire
Songwei SUN ; Yu WU ; Zitong JI ; Rui YANG ; Yufang GAO
Chinese Journal of Modern Nursing 2022;28(10):1275-1280
Objective:To translate the Treatment Burden Questionnaire (TBQ) into Chinese, and test its reliability and validity.Methods:The original questionnaire was translated, synthesized and back-translated according to the Brislin model. From March to July 2021, convenience sampling was used to select 240 elderly people from the 6 designated nursing homes for the aged of Qingdao Municipal Medical Insurance Bureau as the research object to conduct a questionnaire survey. The content validity index ( CVI) and construct validity were used to evaluate the validity of the scale, and the construct validity was evaluated by exploratory factor analysis. Cronbach's alpha coefficient and test-retest reliability were used to test the reliability of the scale. Results:A total of 240 questionnaires were distributed in this study, and 220 valid questionnaires were recovered. The Chinese version of TBQ included 15 items. A total of three common factors were extracted by exploratory factor analysis, and the cumulative variance contribution rate was 58.15%. The item-level CVI of the Chinese version of the TBQ was from 0.80 to 1.00, and the scale-level CVI was 0.91. The Cronbach's α of the total scale was 0.71, and the test-retest reliability coefficient was 0.77. Conclusions:The Chinese version of the TBQ has good reliability and validity, and can be used to assess the treatment burden in the elderly under the Chinese cultural background.
10.Effects of ascites derived exosomes on the stemness and invasion ability of ovarian cancer stem-like cell
Sipan CHEN ; Yufang XIA ; Huanhuan JIANG ; Wenyu SUN ; Yanhui LOU
Chinese Journal of Obstetrics and Gynecology 2021;56(5):349-357
Objective:To investigate the effects of ovarian cancer ascites-derived exosomes on the stem cell properties and invasion ability of ovarian cancer stem-like cell (OCS-LC).Methods:(1) A2780 cells were induced into OCS-LC in serum-free medium, and authenticating their stem-like properties by sphere-forming test, differentiation test and CD 133 marker detection. (2) Exosomes from ascites and A2780 cell were extracted by ultracentrifugation, then authenticating them. The morphology of exosomes was observed by the transmission electron microscope, exosome particle size was measured by nanoparticle tracking analysis (NTA). The expressions of heat shock protein 70 (HSP-70), CD 63 and CD 9 were detected by western blot. (3) The exosomes from ovarian cancer ascites and tumor cell supernate were co-cultured with OCS-LC. The groups were divided into control group, ascites-derived exosomes (ADE) group (ADE+OCS-LC group), and cells-derived exosomes (CDE) group (CDE+OCS-LC group). The sphere-forming ability was evaluated by sphere-forming cycle, maximum sphere diameter and sphere-forming rate of each group; the expression of CD 133 was detected by immunofluorescence staining under microscope; quantitative real-time (qRT)-PCR was used to detected the expression levels of octamer-4 (Oct-4), Nanog mRNA of the signature genes in the stem cells of each group; the metastasis ability of each group was measured by transwell assay. Results:(1) Identification of OCS-LC: sphere-forming experiment showed that the suspension of OCSC single cells was grown in serum-free medium in secondary sphere-forming. Differentiation function experiment showed that OCS-LC were differentiated into adherent A2780 cells by changing the growth mode in serum containing medium. Flow cytometry showed that the proportion of CD 133 positive ( CD133+) cells in OCS-LC group was (18.9±0.9)%, significantly higher than that of control group (0.6±0.5)% ( t=38.570, P<0.01). (2) Under transmission electron microscope, clear lipid bilayer structure was observed in ADE and CDE, and one side presented a concave hemispheric or cup like structure. NTA showed that the diameter of exosomes mainly ranged from 30 to 100 nm, with an average diameter of 67.2 nm. Western blot analysis showed that the specific protein molecules HSP-70, CD 63 and CD 9 were positive. (3) Three groups′ OCS-LC could continue to grow into spheres, and the group of ADE+OCS-LC showed two growth modes, most of the cells continued to grow into spheres, while a small part of cells grew in adherent differentiation. The sphere-forming rate of OCS-LC in the control group, ADE+OCS-LC group, and CDE+OCS-LC group were (1.05±0.20)%, (4.15±0.10)%, and (10.45±0.25)%, the sphere-forming cycle of OCS-LC in the three groups were (15.3±1.5), (10.3±0.6), and (6.7±0.6) days, and the maximum diameters of OCS-LC were (100.3±3.2), (145.2±5.1) and (170.0±2.1) μm, respectively. And the differences were statistically significant (all P<0.05). After co-culture of exosomes with OCS-LC, the sphere-forming ability of cells in the group of CDE+OCS-LC was prior to ADE+OCS-LC group (all P<0.05). Immunofluorescence staining showed that the number of CD 133 green fluorescent chromophore cells in OCS-LC groups [(46.2±2.1)%, (58.4±2.2)%] was significantly higher than that in the control group [(26.6±1.5)%] after the addition of exosomes in co-culture, the positive rate of CD 133 was higher than that in the control group( F=187.588, P<0.05). The qRT-PCR results showed that the expression levels of Oct-4 mRNA in ADE+OCS-LC and CDE+OCS-LC groups were 3.46±0.24, 4.03±0.31, compared with that in control group (1.04±0.12), the differences were statistically significant ( F=134.932, P<0.05). The mRNA expression levels of Nanog were 1.57±0.32, 2.66±0.15, which were significantly higher than that in the control group (1.00±0.07), and the differences were statistically significant ( F=49.329, P<0.05). And the expression of both in CDE+OCS-LC group increased more significantly than ADE+OCS-LC group (all P<0.05). The number of invasive cells in the three groups of OCS-LC were: control group 30±5, ADE+OCS-LC group 102±4, CDE+OCS-LC group 210±7, and there were statistically significant differences among three groups ( F=820.800, P<0.05). Compared with the control group, the number of invaded cells in the co-culture group were significantly increased ( P<0.05), and the CDE+OCS-LC group had the higher cell invasion ability then the ADE+OCS-LC group ( t=23.202, P<0.05). Conclusions:Exosomes derived from ovarian cancer ascites could enhance and maintain the stemness of OSC-LC, and promote the invasion of tumor cells. Moreover, CDE is superior to ADE.

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