1.Construction of lentiviral vector encoding CLEC4M and overexpression of CLEC4M in K-562 cells
Yuanyuan WANG ; Qinghe NIE ; Ting ZHU
Journal of Clinical Hepatology 2014;30(6):518-521
Objective To construct the lentiviral vector encoding CLEC4M and prepare K -562 cells with stable overexpression of CLEC4M.Methods The gene sequence of normal CLEC4Mwas cloned by reverse transcription PCR and then inserted into GV166 vector to construct GV166-CLEC4Mlentiviral expression vector,and then lentiviral packaging was performed by transfection of293T cells.The ob-tained lentiviral liquid was used to infect human leukemia cell line K-562.Real-time PCR and Western blot were used to detect the over-expression of CLEC4M in K-562 cells.Results Sequencing showed that the recombinant lentiviral expression plasmid GV166-CLEC4M was successfully constructed.Lentiviruses could efficiently infect K-562 cells,according to real-time PCR.CLEC4Mwas successfully o-ver-expressed in K-562 cells at mRNA and protein levels.Conclusion The construction of lentiviral vector encoding CLEC4M lays a foundation for further study of CLEC4M gene involved in HCV entry into host cells.
2.Mechanisms of paeoniflorin on T cells in Bovine Collagen-induced arthritis model
Li CAI ; Han ZHANG ; Yuanyuan QI ; Lili ZHANG ; Xiaowan HUANG ; Hong NIE
Chinese Journal of Immunology 2014;(11):1485-1489
Objective:To investigate the mechanisms of paeoniflorin on T cells in Bovine Collagen-induced arthritis ( CIA) model.Methods:CIA were established with Bovine Type Ⅱ collagen in DBA/1J mice.From day 21,after divided into two groups at random,mice were treated with PBS or paeoniflorin and recorded arthritis scores every day;the inflammation of CIA was evaluated with HE staining of the joints;the proliferations of splenocytes and CD4+T cells stimulated either by CD3 antibody and CD28 antibody or CII were examined by 3 H-TdR incorporation assay;the ratio of Th1 and Th17 subsets in CD4+T cells were examined by flow cytometry;the concentration of IFN-γand IL-17 in the culture supernatants of splenocytes were detected by ELISA.Results: The administration of paeoniflorin alleviated the symptoms( P<0.05) and inhibited the inflammation response of CIA;paeoniflorin not only inhibited the pro-liferation of CD3/28 antibodies-stimulated splenocytes and CD4+T cells(P<0.05),but also inhibited the proliferation of CII-specific splenocytes and CII-specific CD4+T cells(P<0.01);paeoniflorin modulated the ratio of Th1 and Th17 subsets(P<0.01);paeoniflorin reduced the levels of IFN-γand IL-17 in the culture supernatant of splenocytes ( P<0.05 ).Conclusion: Paeoniflorin may alleviates clinical symptoms of CIA via inhibiting pathogenic T cell proliferation, balancing inflammatory T cell subsets and reducing pro-inflammatory cytokines.
3.Comparison of rectal cancer tumor volume parameters measured by MRI sequences and CT with those by pathological specimen
Yuanyuan LIU ; Suyu ZHU ; Xiaoyan CHEN ; Lu WEN ; Jinjiao LI ; Xujie BAO ; Jumei ZHOU ; Shaolin NIE
Chinese Journal of Clinical Oncology 2017;44(13):656-661
Objective:This study aimed to compare rectal cancer tumor volume parameters measured by MRI sequences (T1WI, T2WI, and DWI) and/or CT with those by pathological specimen. Methods:Twenty-two patients with rectal cancer were prospectively enrolled. MRI sequences including T1WI, T2WI, and DWI, and/or CT of the pelvis were performed before operation. Volume parameters, such as tumor length along the rectal axis, maximum tumor width perpendicular to rectal axis, and tumor actual area in that perpendicular plane, were measured on T1WI, T2WI, DWI, and CT, respectively, for each patient. The respective pathological parameters were further measured in surgical specimen after total mesorectal excision. The two kinds of parameter values measured in imaging and pathology were statistically compared and accuracy appraisal was performed. Results:The mean Lpath-L was 4.06±1.14 cm. The mean LT1-L, LT2-L, LDWI-L, and LCT-L were 3.91± 1.51, 4.62±1.41, 3.39±1.05, and 3.94±1.23 cm, respectively. Correlation coefficients were 0.688, 0.635, 0.688, and 0.720 (P<0.05). An average 6 mm overestimation was found in T2WI, and 1 to 6 mm underestimation in T1WI, DWI, and CT in length values compared with those measured in surgical specimen. The mean Lpath-W was 2.56 ±0.94 cm. The mean LT1-W, LT2-W, LDWI-W, and LCT-W were 3.62±0.99, 3.66±0.76, 3.23±0.58, and 3.64±1.04 cm, respectively. The magnitude of mean overestimation ranged from 5.1 to 11.1 mm. The Apath was 4.30 ±2.83 cm2. AT1, AT2, ADWI, and ACT were 8.98±3.90, 8.99±3.43, 8.41±3.09, and 9.63±4.40 cm2, respectively, which double overestimated the tumor area in the perpendicular rectal plane. Conclusion:The difference in longitudinal length between MRI sequences/CT and pathological specimen was in the range of?6 mm to 6 mm. The mean maximum tumor width and areas in the maximum tumor perpendicular plane were overestimated. This study indicated that gross tumor volume delineation based on CT or MRI for rectal cancer irradiation should be conservative in the axial images of rectum, and meticulous consideration is required along the rectum.
4.Effect of Community-based Rehabilitation Simple Technique on Motor Function and Balance Function of Stroke
Shuang SHAO ; Lijuan YANG ; Yuanyuan LI ; Xiaofeng JIANG ; Lihua NIE ; Hong DAI ; Wenzhi WANG
Chinese Journal of Rehabilitation Theory and Practice 2010;16(4):390-392
ObjectiveTo investigate the effects of community-based rehabilitation simple technique on motor function, balance function of stroke.Methods48 stroke patients from Guangwai Hospital in Beijing were divided into two groups: test group(n=26) and control group(n=22). The test group received community-based rehabilitation simple technique combining with routine therapy, while the control group only received routine therapy. All patients were assessed with the scale of motor function (Fugl-Meyer Assessment, FMA) and Fugl-Meyer Balance Assessment (FMA-B) before, 1 month, and 3 months after treatment respectively.ResultsThere were no significant difference between two groups in FMA and FMA-B (P>0-05) before and 1 month after treatment. The scores of FMA and FMA-B improved significantly (P<0-05) 3 months after treatment.ConclusionCommunity-based rehabilitation simple technique is effective to improve motor function and balance function of stroke patients.
5.Correlation research between nurses′status of career planning and job satisfaction, turnover intention
Lei WANG ; Hong SUN ; Xin GUAN ; Shengxiao NIE ; Jianhua ZHANG ; Yuanyuan HAN
Chinese Journal of Modern Nursing 2015;21(31):3721-3723
Objective To explore the correlation between nurses′ status of career planning and job satisfaction, turnover intention, so as to provide evidence for stabilizing nurse team and reducing nurse lost. Methods A total of 1139 nurses were recruited by multi-stage sampling method from 10 level three class A hospitals and were investigated by nurse′s career planning current status questionnaire, nurse′s job satisfaction questionnaire, nurse′s turnover intention questionnaire. Results The average scores of nurse′s career planning and job satisfaction were (3. 85 ± 0. 50) and (3. 18 ± 0. 89). Fifteen nurses intended to leave the present hospital after 1 year, and 173 nurses were not sure whether to leave present hospitals. The correlation was positive between nurses′career planning and job satisfaction (r=0. 372,P<0. 01), but it was negative between nurses′career planning and turnover intention (r= -0. 114,P<0. 01). Conclusions Nurse managers should strengthen nurse′s career planning, help nurses to make their own career planning, so as to enhance nurses′job satisfaction, reduce nurse flow, and reach win-win of personal and hospital.
6.Protective effect of salvianolic acid B on isolated heart ischemia/reperfusion injury in rats.
Feng GAO ; Guibo SUN ; Xiaoyu REN ; Yuanyuan NIE ; Jing SUN ; Meng QIN ; Xiaobo SUN
China Journal of Chinese Materia Medica 2012;37(3):358-361
OBJECTIVETo study the protective effect and mechanism of salvianolic acid B on isolated heart ischemia/reperfusion injury in rats.
METHODForty-eight SD rats were divided into 6 groups randomly(n = 8): the control group, the positive administration group (verapamil 150 microg x L(-1)), and high, middle and low-dose salvianolic acid B groups (10, 5, 2.5 mg x L(-1)). The myocardial ischemia/reperfusion injury model was established using the Langendorff method, re-perusing isolated working hearts for 30 min after ischemia for 25 min. A water-bag catheter was inserted in rat left atrium for recording the effect of salvianolic acid B on hemodynamics indexes-AST, LDH, SOD and MDA.
RESULTVarious group with different doses showed that salvianolic acid B decreased AST, release of LDH and formation of MDA and increased SOD activity.
CONCLUSIONSalvianolic acid B showed a protective effect on myocardial ischemia/reperfusion injury. Its mechanism may be related with improvement of cardiac contractility, cleaning of oxygen free radicals and reduction of lipid peroxidation.
Animals ; Benzofurans ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Hemodynamics ; drug effects ; In Vitro Techniques ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Time Factors
7.Bibliometrics analysis on identification of molecula pharmacognosyin based on Web of Science
Ying NIE ; Yuanyuan TONG ; Yang LIU ; Haiyan LI
International Journal of Traditional Chinese Medicine 2018;40(2):136-142
Objective To analyze the literature characteristics on molecular identification of pharmacognosy, summarize the current research situation and trend in the field and to provide a reference for relative researchers. Methods The papers relevant to molecular identification included in SCIE were statistically analyzed by biblimetric methods from the aspects of publishing year,journals disbution, author's unit,author's country or region and technique.And the papers also use the visualization techonology to visual the links between the keywords. Results The total number of the literature for analyzing was 5108, the numbers of papers increased gradually.These paper involved 3720 authors, 1695 organizations. and 421 journals. Authors of the papers mainly from China, the United States and Korea. The traditional Chinese medicine involved were Catharanthusroseus, Panax ginseng, Tobacco,wheat, Artemisia annua. The relative identification technology top 5 were RAPD, microsatellite, AFLP, ISSR, HPLC.Based on visiualization technology, we found the current research hotspots were base source identification, habitats identification, population genetics and molecular phylogeography. Conclusions The research of molecular identification of pharmacognosyin were on the rise. The identification technology and species studied will extend ulteriorly.
8.Recurrent stroke in patients w ith right-to-left shunt and cryptogenic stroke
Fang ZHOU ; Aixia ZHUANG ; Shouqin FENG ; Qinghong ZENG ; Yujuan QI ; Le YIN ; Yuanyuan WANG ; Hongxia NIE ; Yang WANG ; Chuanqi WANG ; Shanhua YU ; Jiandong JIANG
International Journal of Cerebrovascular Diseases 2015;(5):344-348
Objective To investigate the relationship between the right-to-left shunt(RLS)detected with contrast-enhanced transcranial Doppler (c-TCD) and recurrent stroke in patients with cryptogenic stroke.Methods The consecutive patients w ith ischemic stroke w ere enrol ed. The patients w ith cryptogenic stroke w ere screened according to the TOAST criteria. They w ere divided into either a RLS positive group or a RLS negative group according to the c-TCD findings, and then they w ere fol ow ed up for a period of one year. They w ere also divided into a recurrent group and a non-recurrent group according to w hether they had recurrence or not. Results A total of 118 patients w ith cryptogenic ischemic stroke w ere enrol ed, including 46 in the RLS positive group, 72 in the RLS negative group, 10 in the recurrent group, and 108 in the non-recurrent group. There w ere no significant differences in demographic and baseline data betw een the RLS negative group and the RLS positive group. There w ere significant differences in RLS positive rate (7/10 vs.39/108; P=0.046) and proportion of patients with server RLS (2/10 vs.1/108; P=0.019) betw een the recurrent group and the non-recurrent group. Multivariate logistic regression analysis show ed that the positive RLS w as an independent predictor of recurrent stroke (odds ratio 4.896, 95% confidence interval 1.135-21.120;P=0.033). Conclusions The positive RLS may be an independent risk factor for the recurrence in patients w ith cryptogenic ischemic stroke.
9.Serum exosome-mediated lncRNA HOTTIP regulates cisplatin resistance in gastric cancer cells through the miR-138-5p/TJP1 axis
Mingyang HAN ; Junwei BAI ; Jiewei NIE ; Yuanyuan LI ; Chao ZHANG
Chinese Journal of Digestive Surgery 2023;22(12):1467-1475
Objective:To investigate the effects of serum exosomes of patients with gastric cancer on cisplatin resistance, clonal formation, migration, invasion and apoptosis of the AGS gastric cancer cells, and the corresponding molecular mechanisms.Methods:The experimental study was conducted. The exosomes of patients with gastric cancer was separated from their serum, and the expression of lncRNA HOTTIP was analyzed using the quantitative real time polymerase chain reaction (qRT-PCR). Normal gastric epithelial cell line GES1, gastric cancer cell line AGS and human embryonic kidney cell 293T were cultured in vitro. AGS cells were incubated with exosomes (Exo),with phos-phate buffered saline (PBS) treatment as control, and transfected with si-NC or si-HOTTIP-3, named as Exo group, PBS group, si-NC+Exo group, and si-HOTTIP-3+Exo group. The AGS cells were trans-fected with si-NC, si-HOTTIP-1, si-HOTTIP-2, si-HOTTIP-3, oe-HOTTIP, vector, oe-HOTTIP+miR-138-5p mimic, oe-HOTTIP+mimic NC, miR-138-5p inhibitor, inhibitor NC, miR-138-5p inhibitor+si-TJP1 and miR-138-5p inhibitor+si-NC. They were recorded as si-NC group, si-HOTTIP-1 group, si-HOTTIP-2 group, si-HOTTIP-3 group, oe-HOTTIP group, vector group, oe-HOTTIP+miR-138-5p mimic group, oe-HOTTIP+mimic NC group, miR-138-5p inhibitor group, inhibitor NC group, miR-138-5p inhibitor+si-TJP1 group and miR-138-5p inhibitor+si-NC group. The 293T cells transfected with mimic NC+HOTTIP wt, miR-138-5p mimic+HOTTIP wt, mimic NC+HOTTIP mut, miR-138-5p mimic+HOTTIP mut, mimic NC+TJP1 3'UTR wt, miR-138-5p mimic+TJP1 3'UTR wt, mimic NC+TJP1 3'UTR mut, miR-138-5p mimic+TJP1 3'UTR mut were recorded as the mimic NC+HOTTIP wt group, miR-138-5p mimic+HOTTIP wt group, mimic NC+HOTTIP mut group, miR-138-5p mimic+HOTTIP mut group, mimic NC+TJP1 3'UTR wt group, miR-138-5p mimic+TJP1 3'UTR wt group, mimic NC+TJP1 3'UTR mut group, miR-138-5p mimic+TJP1 3'UTR mut group. The cell counting kit-8 (CCK8) was used to analyze the cisplatin sensitivity of gastric cancer cells. The colony formation experiment was used to analyze the colony formation of gastric cancer cells. The Transwell experiment was used to analyzed cell migration and invasion of gastric cancer cells. The flow cytometry experiment was used to analyze cell apoptosis of gastric cancer cells. The Western bolt assay was used to analyze the expression of exosome marker proteins, including the CD63 and CD81, and the protein of TJP1, the drug-resistance related proteins, including the P-gp and MCL-1. The dual-luciferase assay was used to analyze the targeted relationships among lncRNA HOTTIP, miR-138-5p and TJP1. Observation indicators: (1) expression of lncRNA HOTTIP; (2) resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP; (3) regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression; (4) targeting of TJP1 gene 3′-untranslated region (UTR) by miR-138-5p; (5) regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. The one-way ANOVA was used for comparison for multiple groups and the Tukey′s test was used for further pairwise compari-son. Count data were described as absolute numbers, and the chi-square test was used for comparison. Correlation analysis was conducted using the Pearson′s test. Results:(1) Expression of lncRNA HOTTIP. The expression of lncRNA HOTTIP in the serum exosome of patients with gastric cancer was higher than that in healthy volunteers, showing a significant difference ( P<0.05). Results of transmi-ssion electron microscopy examination showed that the serum exosomes were circular or oval in shape. Results of Western bolt assay showed the expression of marker proteins of CD63 and CD81 in serum exosomes. (2) Resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP. Compared with the PBS group, the biochemical half maximal inhibitory concentra-tion (IC50), the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the Exo group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the si-NC+Exo group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the si-HOTTIP-3+Exo group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). (3) Regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression. Compared with the vector group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the oe-HOTTIP+mimic NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP+miR-138-5p mimic group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). (4) Targeting of TJP1 gene 3′-UTR by miR-138-5p. Results of dual-luciferase assay showed that the luciferase activity in 293T cells treatment with mimics of control+vectors of wild type of TJP1 gene 3′-UTR and 293T cells treatment with mimics of miR-138-5p+vectors of wild type of TJP1 gene 3′-UTR was 1.00±0.09 and 0.21±0.03, respectively, showing a significant difference between them ( t=15.02, P<0.05). (5) Regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Compared with the inhibitor group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the miR-138-5p inhibitor+si-NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor+si-TJP1 group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). Conclusion:Serum exosomes-mediated lncRNA HOTTIP can promote cisplatin resistance, clonal formation, migration, invasion and apoptosis of gastric cancer cells and inhibit cell apoptosis of gastric cancer cells through regulating the expression of miR-138-5p/TJP1.
10.A survey of inflammatory bowel disease in China based on the Google Trends, Baidu index and WeChat index
Kai NIE ; Xiaoyan WANG ; Kejia MA ; Yuanyuan YANG ; Minzi DENG
Chinese Journal of Digestion 2022;42(6):395-401
Objective:To analyze the epidemiological trends and population characteristics of inflammatory bowel disease(IBD)in China by searching keywords related to IBD through Google Trends, Baidu index, and WeChat index, so as to provide a reference for national epidemiological studies on IBD.Methods:IBD-related hot words such as "inflammatory bowel disease" , "Crohn′s disease" , "ulcerative colitis" , "gastroesophageal reflux disease (GERD)" , "irritable bowel syndrome (IBS)" and " Helicobacter pylori ( H. pylori)" were selected. The search volume and trends of the above keywords in the world and China were analyzed through Google Trends, Baidu index and WeChat index. The epidemiological characteristics of IBD in China were summarized. Descriptive methods were used for statistical analysis. Results:The results of Google Trends analysis showed that among 5 common digestive diseases (GERD, IBS, H. pylori infection, IBD and peptic ulcer), GERD was the most concerned disease, while IBD was not the focus among the common digestive diseases. When the global searching scope was limited to IBD related hot words, Crohn′s disease was the disease of primary concern among IBD-related diseases. In South America, South Asia, and parts of Africa, ulcerative colitis was mainly concerned, and in China and countries of Southeast Asia, IBD was more concerned. The searching results of Baidu index indicated that among the national search for IBD-related hot words, the 3 keywords of "inflammatory bowel disease" , "ulcerative colitis" and "Crohn′s disease" were all highly searched, the overall daily average of the search indexes of the 3 keywords were 325, 1 320 and 2 559, respectively, and the searching volume of "Crohn′s disease" was higher than "inflammatory bowel disease" and "ulcerative colitis" . The national wide trends of search volume for "inflammatory bowel disease" , "ulcerative colitis" and "Crohn′s disease" were similar, the search heat gradually decreased from the east coast to the northwest of China, which basically coincided with the three-level ladder trend of China′s economic development, suggesting that the level of economic development was related to the incidence of IBD. The results of Baidu index analysis showed that the main populations who searched IBD-related keywords were young adults and women aged from 20 to 39 years old. The results of WeChat index analysis revealed that the searching volume of "inflammatory bowel disease" , "ulcerative colitis" and "Crohn′s disease" were 205 000, 195 000 and 120 000, respectively, and the search volume for " inflammatory bowel disease" was the highest. The official account (90.27%) and the video account (7.43%) occupied the main sources of IBD-related information on mobile terminals. Conclusions:The IBD-associated internet activities reveal a global lack of public awareness of IBD. China also faces the same problem. The search trend is consistent with the epidemiology of IBD, which may be helpful for future epidemiological research of IBD in China. Mobile media will be a potential force in promoting the patient education and disease management of IBD in China.