1.miR-23a Regulates Proliferation and Apoptosis of Rectal Cancer via Targeting Gene ESRP1.
Lingjuan LAO ; Xinjiang SONG ; Jia XU
Journal of Medical Research 2017;46(6):121-125
Objective To elucidate the relative level of miR-23a RNA in rectal cancer tissues and cell line as well as the effects of miR-23a on the cell proliferation and apoptosis of rectal cancer cells in vitro.Methods Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was applied in assessment of the transcription of miR-23a in rectal cancer tissues and in vitro cells.The RNA fragment of miR-23a inhibitor and inhibitor NC were synthesized and transfected into SW480 cells.Cell proliferation was evaluated with Cell Counting Kit-8 (CCK-8) assay.The apoptotic rate was analyzed by flow cytometry.The expression of ESRP1 was detected by western blot.Wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ES-RP1-3'UTR) plasmids and miR-23a inhibitor RNA fragments or inhibitor NC RNA fragments were co-transfected into HEK293 and SW480 cells,then the Promega dual luciferase reporter gene assay kit was used to examine the dual luciferase activity in SW480 cells.Resuits The relative RNA level of miR-23a was significantly promoted in both rectal cancer tissue samples and SW480 cells.After SW480cells were transfected with miR-23a inhibitor,human rectal cancer cell line SW480 with down-regulation of miR-23a showed significant inhibition of cell proliferation compared with negative control (P =0.000).Furthermore,our data demonstrated clearly that the inhibition of miR-23a promoted apoptosis in SW480 cells (P =0.000).Luciferase assay showed that ESRP1 was a direct target gene of miR -23a.Conclusion The expression of miR-23a is clearly associated with the growth and apoptosis of human rectal cells by targeting ESRP1,whilst miR-23a may be used as a potential therapeutic target for the treatment of rectal cancer in the future.
2.miR-23a regulates cell growth and apoptosis of rectal cancer via targeting ESRP1 gene
Lingjuan LAO ; Xinjiang SONG ; Jia XU
Chinese Journal of Pathophysiology 2017;33(5):817-825
AIM:To investigate the expression of miR-23a and epithelial splicing regulatory protein 1(ESRP1) in rectal cancer tissues and cell lines as well as their effects on rectal cancer cell viability and apoptosis.METHODS:The relative levels of miR-23a in the rectal cancer tissues and cultured cells were assessed by RT-qPCR.The positive expression of ESRP1 in the rectal cancer tissues and non-cancer tissues was detected by immunohistochemical staining.The sequences of miR-23a inhibitor and inhibitor negative control (NC) were synthesized, and transfected into the SW480 cells.The cell viability was measured by CCK-8 assay.The apoptotic rate was analyzed by flow cytometry.The cell invasion was evaluated by Matrigel counting assay.The expression of ESRP1 was determined by Western blot.The wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ESRP1-3'UTR) plasmid and miR-23a inhibitor or inhibitor NC were co-transfected into the HEK293 and SW480 cells.The dual luciferase activity was detected according to Promega dual luciferase reporter gene assay kit instructions.The cell viability and apoptosis were evaluated by CCK-8 assay and flow cytometry analysis, respectively, after the SW480 cells were transfected with ESRP1 mimic or mimic NC.The expression of ESRP1, caspase-3, Smac and X-linked inhibitor of apoptosis protein (XIAP) in the SW480 cells was detected by Western blot.RESULTS:The expression of miR-23a was significantly up-re-gulated in the rectal cancer tissues and cell lines, while the positive expression of ESRP1 was significantly decreased in the rectal cancer specimens.The miR-23a expression was also closely related to lymphnode metastasis and TNM stages of rectal cancer patients.ESRP1 was inversely correlated with miR-23a in the rectal cancer tissues.After transfection with miR-23a inhibitor in human rectal cancer SW480 cells, the down-regulation of miR-23a induced significant inhibition of cell viability as compared with the cells transfected with inhibitor NC (P<0.01).Furthermore, the apoptotic rate induced by the miR-23a inhibitor transfection was markedly higher than that of control (P<0.01).Luciferase assay showed that ESRP1 was a direct target gene of miR-23a.The cell viability and apoptosis were inhibited and promoted, respectively, after transfection with ESRP1 mimic in the SW480 cells.Promoted expression of ESRP1 significantly up-regulated the levels of caspase-3 and Smac as well as down-regulated the expression of XIAP in the SW480 cells.CONCLUSION:The expression of miR-23a is significantly associated with the growth and apoptosis of human rectal cancer cells by targeting ESRP1.miR-23a may be a potential therapeutic target for the treatment of rectal cancer in the future.
3.Necessity of central lymph node dissection in management of papillary thyroid microcarcinoma.
Xinjiang XU ; Bin JIANG ; Liang HAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2014;28(6):362-365
OBJECTIVE:
The objective of this study was to identify the risk factors for central lymph node metastasis (CLNM) of papillary thyroid microcarcinoma(PTMC) and to explore the necessity of central lymph node dissection (CLND).
METHOD:
Clinical data of 85 patients with PTMC, who had undergone surgical treatment between January 2004 and May 2012, were retrospected. Risk factors for CLNM were identified by univariate analysis and multivariate analysis,which can provide the basis for elective performance of CLND.
RESULT:
Of 85 patients,66 patients underwent ipsilateral CLND,while 19 patients received bilateral CLND. Concurrent cervical lymph node dissection was performed in 3 patients. The incidence of central and cervical lymph node metastasis was 38.8% and 3.53%, respectively. Univariate analysis showed that CLNM was correlated with tumor size > 5 mm, extrathyroidal extension, multifocality, bilaterality and intraoperatively suspected lymph node, but not related to gender and age. Upon multivariate analysis, tumor size > 5 mm (OR = 3.862, P < 0.05) and extrathyroidal extension (OR = 3.885, P < 0.05) were independent risk factors for CLNM.
CONCLUSION
Patients presenting tumor size > 5 mm and/or extrathyroidal extension may have an increased risk of central lymph node metastasis,and it is necessary to perform central lymph node dissection for them.
Adult
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Aged
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Carcinoma, Papillary
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secondary
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surgery
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Elective Surgical Procedures
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Female
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Humans
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Incidence
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Lymph Node Excision
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Lymph Nodes
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Lymphatic Metastasis
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Male
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Middle Aged
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Multivariate Analysis
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Retrospective Studies
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Risk Factors
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Thyroid Neoplasms
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secondary
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surgery
4. Effect of different concentrations of curcumin on inflammatory factors and endotoxin changes in lung tissues of rats with heatstroke in dry-heat environment
Jia-jia LI ; Na MA ; Xiang DONG ; Wei CAO ; Jing JIANG ; Yan KANG ; Jian-ying LI ; Lai-yang SONG ; Qin XU ; Jiang-wei LIU
Journal of Medical Postgraduates 2018;31(12):1258-1261
Objective Thermal injury causes pulmonary edema, which may lead to acute respiratory distress syndrome, sepsis, multiorgan failure and even death. The article aimed to study the mechanism of curcumin pretreatment on inflammatory factors in lung tissues and serum endotoxin of rats with dry-heat environment.Methods A total of 50 SD rats were randomly divided into 5 groups (n=10): normal control group, dry heat control group, low concentraion group (50mg/kg curcumin pretreatment group), middle concentraion group (100mg/kg curcumin pretreatment group), and high concentration group (200mg/kg curcumin pretreatment group). Rats in normal control group and dry heat control group were given normal saline by gavage, while rats in 3 curcumin pretreatment groups were given curcumin of different concentrations (50 mg/kg, 100 mg/kg, 200 mg/kg) by gavage once a day for 7 consecutive days. At 8d, all the other 4 groups except normal control group were transferred to the climate cabin (The Simulated Climate Cabin for Special Environment of Northwest of China) with the condition of (41±0.5)℃, (10±1)% relative humidity.The rats were put in the dry-heat environment for 150min, then they were anaesthetized and sacrificed at 150min to collect the blood, lung tissues for further analysis. Observation was made on the pathological changes of lung tissues of rats in each group and the changes of inflammatory factors such as IL-1β, IL-6, TNF-α and LPS.Results Compared with dry heat control group, the concentrations of IL-1β, IL-6, TNF-α and LPS in normal control group, curcumin pretreatment groups with low concentration, middle concentration and high concentration were significantly higher(P<0.01). The concentrations of IL-1β, IL-6, TNF-α and LPS in curcumin pretreatment group with low concentration were significantly lower than those curcumin pretreatment groups with middle concentration and high concentration(P<0.05). Compared with curcumin pretreatment group with middle concentration, LPS concentration of curcumin pretreatment group with high concentration decreased significantly (P<0.01). Pearson correlation analysis showed that there was a positive correlation between the plasma of LPS and inflammatory cytokines of IL-1β, IL-6, and TNF-α in lung tissues (correlation coefficient r=0.866, r=0.900, r=0.885, P=0.000).Conclusion Curcumin inhibits bacterial endotoxin in blood, reduces the expression of inflammatory factors, and plays an important role in alleviating secondary multiple organ damage, which means curcumin pretreatment can relieve lung damage caused by heatstroke and reduce the mortality of heatstroke.
5.Recent advances and applications of base editing systems.
Chinese Journal of Biotechnology 2021;37(7):2307-2321
The CRISPR system is able to accomplish precise base editing in genomic DNA, but relies on the cellular homology-directed recombination repair pathway and is therefore extremely inefficient. Base editing is a new genome editing technique developed based on the CRISPR/Cas9 system. Two base editors (cytosine base editor and adenine base editor) were developed by fusing catalytically disabled nucleases with different necleobase deaminases. These two base editors are able to perform C>T (G>A) or A>G (T>C) transition without generating DNA double-stranded breaks. The base editing technique has been widely used in gene therapy, animal models construction, precision animal breeding and gene function analysis, providing a powerful tool for basic and applied research. This review summarized the development process, technical advantages, current applications, challenges and perspectives for base editing technique, aiming to help the readers better understand and use the base editing technique.
Adenine
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Animals
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CRISPR-Cas Systems/genetics*
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Cytosine
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DNA Breaks, Double-Stranded
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Gene Editing
6. Influence of job burnout and sleep quality on working ability of oil workers
Xue LI ; Xu YANG ; Ji-wen LIU
China Occupational Medicine 2021;48(03):266-271
OBJECTIVE: To explore the current situation of job burnout, sleep quality and working ability and the association among them in oil workers. METHODS: A total of 2 086 oil workers from six oil field companies in a city of Xinjiang Uygur Autonomous Region were selected as the study subjects using a typical sampling method. The job burnout level, sleep quality and working ability were measured respectively by the Maslach Burnout Inventory-General Survey, Pittsburgh Sleep Quality Index and Work Ability Index Questionnaire. RESULTS: The average score of job burnout in the study subjects was(48.6±11.0), the median and the 25 th, 75 th percentile of sleep quality and working ability scores were 6.0(5.0, 8.0) and 39.0(36.0, 42.0) respectively. There was a negative correlation between job burnout and working ability, or sleep quality and working ability [spearman correlation coefficient(r_S) were-0.28 and-0.21, all P<0.01]. There was a positive correlation between job burnout and sleep quality(r_S=0.19, P<0.01). The structural equation model results showed that both job burnout and sleep quality had a direct effect on working ability of oil workers [normalized path coefficient(β) were-0.29 and-0.27, respectively, all P<0.01]. Job burnout could indirectly affect working ability through reducing sleep quality(β=-0.12, P<0.01). CONCLUSION: The higher the degree of job burnout and the poorer the sleep quality, the lower their working ability in oil workers. Oil workers with a higher degree of job burnout can lead to poorer sleep quality, which in turn leads to a decline in their working ability.
7.Influence of Guizhi Jia Longgu Mulitang on Expression of IL-10 and TNF-α in Insomnia Rats with Sensory Dysfunction Dominated by Lung
Jinhong WU ; Xingping ZHANG ; Deqi YAN ; Ruining LIANG ; Xu CHEN ; Zhengting LIANG ; Honglin JIA ; Hui WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):20-27
ObjectiveTo investigate the effects of Guizhi Jia Longgu Mulitang on the expression difference of interleukin-10 (IL-10) and tumor necrosis factor -α (TNF-α) in related organs of insomnia rats with sensory dysfunction dominated by lung and study the mechanism of Guizhi Jia Longgu Mulitang in improving insomnia. MethodSD rats were randomly divided into the blank group, model group, western medicine group, and traditional Chinese medicine (TCM) group, with 10 rats in each group. The rats were deprived of sleep by shallow water environment method in a long platform, and the modeling lasted for 42 d. The blank group and model group were given 0.05 mL·kg-1 normal saline by gavage, and the western medicine group and TCM group were given drugs during modeling. To be specific, the western medicine group was given 0.105 mg·kg-1 dexzopiclone tablet by gavage, while the TCM group was given 7 600 mg·kg-1 Guizhi Jia Longgu Mulitang by gavage, both lasting for 28 days. After successful modeling, the Morris water maze experiment was performed on the 42nd day to detect the motion and spatial memory ability of rats. The levels of IL-10 and TNF-α in serum were detected by enzyme-linked immunosorbent assay (ELISA). The protein expression of IL-10 and TNF-α in the lung and brain tissue of rats was detected by Western blot. The levels of IL-10 and TNF-α in the lung and brain tissue of rats were detected by immunohistochemistry. ResultCompared with the blank group, the sleep stages non-rapid eye movement ( NREM ) and rapid eye movement ( REM ) of the model group were significantly shortened (P<0.5, P<0.01), and the wake stage was significantly increased (P<0.01). The total time and distance of platform exploration were significantly increased (P<0.01). In the target quadrant (the third quadrant), the percentage of exploration time and the times of crossing the platform were significantly decreased (P<0.01). ELISA results showed that the serum IL-10 level was significantly decreased (P<0.01), and TNF-α level was significantly increased (P<0.01). The results of Western blot showed that the protein expression of IL-10 in brain and lung tissue of rats was significantly decreased (P<0.01), and the protein expression of TNF-α was significantly increased (P<0.01). The results of immunohistochemistry showed that the expression of IL-10 in the brain and lung tissue of rats was significantly decreased (P<0.01), and that of TNF-α was significantly increased (P<0.01). Compared with the model group, the NREM stage and REM stage of the western medicine group and the TCM group were significantly increased (P<0.5, P<0.01), and the wake stage was shortened (P<0.5). The total time and distance of platform exploration were significantly decreased (P<0.01). In the target quadrant (the third quadrant), the percentage of exploration time and the times of crossing the platform were significantly increased (P<0.05, P<0.01). The serum IL-10 level was significantly increased (P<0.01), and the serum TNF-α level was significantly decreased according to the ELISA results (P<0.01). The results of Western blot showed that the protein expression of IL-10 in brain tissue and lung tissue was significantly increased (P<0.01), and the protein expression of TNF-α was significantly decreased (P<0.01). The results of immunohistochemistry showed that the expression of IL-10 in brain tissue and lung tissue was significantly increased (P<0.05, P<0.01), and the expression of TNF-α was significantly decreased (P<0.05, P<0.01). ConclusionGuizhi Jia Longgu Mulitang can improve the expression of IL-10 and TNF-α in brain and lung tissue of insomnia rats with sensory dysfunction dominated by lung, prolong sleep time, and then improve insomnia. The mechanism may be related to improving the expression level of inflammatory factors.
8.Research progress of long-term efficacy after implantation of implantable collamer lens V4c
Miaomiao WANG ; Huixian WANG ; Xu ZHANG ; Wenjing LI
International Eye Science 2024;24(2):260-263
With the rapid development of science and technology and the wide application of electronic products, the number of patients with high myopia is gradually increasing. Meanwhile, owing to the continuous optimization of surgical skils and the continuous advancement in materials of posterior chamber intraocular lens and manufacturing processes, implantable collamer lens(ICL)V4c implantation has gradually become one of the main surgeries for the treatment of high myopia. In the rapid era of social information, people pay more attention to the long-term efficacy after ICL V4c implantation, they not only want clear vision, but also the pursuit of visual comfort and durability. Therefore, this paper will specifically discuss the research progress of the post-implantation efficacy of ICL V4c with at least 2 a of follow-up observation within 3 a, aiming to review the latest research progress on the long-term efficacy of ICL V4c implantation from the three dimensions of visual quality, safety, efficacy and stability and possible surgical complications after ICL V4c implantation.
9.Preparetion of anti-CD70 nanobody immunotoxin fused with PE38 and its killing effect on renal clear cell carcinoma 786-O cells
XU Xinlan ; LIU Chang ; ZHANG Xin ; HU Qianqian ; LI Jiangwei
Chinese Journal of Cancer Biotherapy 2023;30(8):665-671
[摘 要] 目的:构建靶向CD70分子的重组免疫毒素,通过表达、纯化制备PE38与抗CD70纳米抗体重组蛋白,体外抗肿瘤实验探究重组蛋白是否对高表达CD70分子的阳性肿瘤细胞具有杀伤活性。方法:通过基因工程手段,将CD70纳米抗体Nb 2B3基因片段通过一个连接子与pET21a-PE38基因片段相连,获得重组表达载体pET21a-Nb 2B3-PE38并转入BL21(DE3)感受态细胞中进行表达、纯化与鉴定。用间接ELISA及FACS法检测Nb 2B3-PE38与CD70分子的结合活性,MTT法检测Nb 2B3-PE38对高表达CD70分子的肾透明细胞癌786-O细胞的体外杀伤活性,Annexin Ⅴ-FITC/PI双染法检测Nb 2B3-PE38对786-O细胞凋亡的影响。结果:成功构建抗CD70纳米抗体重组免疫毒素Nb 2B3-PE38,纯化获得纯度>90%的重组蛋白,SDS-PAGE及WB检测结果表明目的蛋白正确表达,分子量为56 000。纯化后的Nb 2B3-PE38能与重组CD70抗原及786-O细胞表面的CD70分子特异性结合;25 µg/mL Nb 2B3-PE38即对786-O细胞产生极显著的杀伤作用(P<0.001),并且促进786-O细胞的细胞凋亡(P<0.01),其杀伤效应强于阳性对照顺铂(P<0.01)。结论:成功制备了特异性靶向CD70分子的免疫毒素Nb 2B3-PE38,其能够有效杀伤786-O细胞并诱导细胞凋亡且效果强于顺铂。
10.Optimal waist circumference cutoff for categorization of central adiposity among Han adults in Xinjiang and its predictive ability for cardiovascular disease risk factors
Li XU ; Jianfeng YAN ; Shu FANG ; Jie GAO
China Modern Doctor 2024;62(8):6-11
Objective To explore the optimal cutoff for waist circumference(WC)for defining central obesity,and its predictive ability for cardiovascular disease risk factors in Han adults in Xinjiang.Methods A cardiovascular risk survey(CRS)study was carried out from October 2018 to March 2021 with 14 618 representative participants selected,including 5757 Han adults.Complete data on 5666 participants were gathered.The age span of the participants was from 35 to 88 years old with the mean age as 52.47 years.Anthropometric data,blood pressure,serum concentration of serum total cholesterol,triglyceride,low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and fasting glucose were documented.Prevalence rate,sensitivity,specificity and distance on the receiver operator characteristic(ROC)curve of each WC value were calculated.Results Both men and women,with the increase of waist circumference,the levels of systolic blood pressure,diastolic blood pressure,total cholesterol,triglycerides and fasting blood glucose are gradually increasing.In men,the prevalence of hypertension and hypertriglyceridemia increased as WC increased.In women,the prevalence of hypertension,diabetes,hypercholesterolemia and hypertriglyceridemia increased as WC increased.The shortest distance in the ROC curves for hypertension,dyslipidemia,diabetes,or≥2 of these risk factors suggested a WC cutoff of 90cm for men and 85cm for women.Conclusion The proportion of central obesity among Han adults in Xinjiang is higher than that of Han people in other regions.It is necessary to use a higher cutoff point value of waist circumference to define central obesity.