1.Rho kinase inhibitor Y27632 promotes survival of human induced pluripotent stem cells during differentiation into functional midbrain dopaminergic progenitor cells in vitro
Yangyang LI ; Jiajia XU ; Chengcheng JIANG ; Zilong CHEN ; Ying CHEN ; Mengjiao YING ; Ao WANG ; Caiyun MA ; Chunjing WANG ; Yu GUO ; Changqing LIU
Journal of Southern Medical University 2024;44(2):236-243
Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.
2.Rho kinase inhibitor Y27632 promotes survival of human induced pluripotent stem cells during differentiation into functional midbrain dopaminergic progenitor cells in vitro
Yangyang LI ; Jiajia XU ; Chengcheng JIANG ; Zilong CHEN ; Ying CHEN ; Mengjiao YING ; Ao WANG ; Caiyun MA ; Chunjing WANG ; Yu GUO ; Changqing LIU
Journal of Southern Medical University 2024;44(2):236-243
Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.
3.Effects of FOCUS-PDCA management on the improvement of disinfection quality of flexible endoscopes
Xiying CHAI ; Chunmei SHAO ; Yaying HAN ; Ying SONG ; Chengcheng LI
Chinese Journal of Modern Nursing 2022;28(3):392-395
Objective:To explore the effects of FOCUS-PDCA management on the improvement of disinfection quality of flexible endoscopes.Methods:From January 2018 to February 2020, a total of 124 flexible endoscopes and 23 nurses in Endoscopy Center of The First People's Hospital of Shangqiu were selected as the research objects by the convenient sampling method. From January 2018 to January 2019, the flexible endoscopes that were disinfected by conventional methods were set as the control group ( n=62) . From February 2019 to February 2020, the flexible endoscopes treated with Focus-PDCA disinfection were set as the observation group ( n=62) . The implementation status of the cleaning and disinfection specifications, the incidence of validity management defects and the quality of disinfection management of flexible endoscopes were compared between the two groups. Results:The observation group's flexible endoscope cleaning, disinfection specifications, and management evaluation scores were higher than those of the control group, and the differences were statistically significant ( P<0.01) . The disinfection and management defects of the flexible endoscope in the observation group were lower than those in the control group, the completeness rate of cleaning and disinfection records, the standard rate of cleaning operations, the qualification rate of biological test results, and the implementation rate of disinfection protection measures in the observation group were all higher than those of the control group, the differences were statistically significant. ( P<0.05) . Conclusions:The FOCUS-PDCA management model can effectively improve the cleanliness and disinfection quality of flexible endoscopes, which is worthy of clinical promotion.
4.HPLC Fingerprint Establishment ,Content Determination and Multivariate Statistical Analysis of Qinlian Runfei Decoction
Kaili YAN ; Chengcheng YIN ; Mengyao LIU ; Changsheng CUI ; Ying WANG ; Xiaozhou SUN ; Liping SUN ; Bin QI ; Li LIU
China Pharmacy 2021;32(16):1956-1963
OBJECTIVE:To establish the f ingerprint of Qinlian runfei decoction,determine the contents of 11 components, and conduct cluster analysis and orthogonal partial least squares discriminant analysis (OPLS-DA). METHODS :HPLC method was used. The determination was performed on ZORBAX Eclipse Plus C 18 column with mobile phase consisted of acetonitrile- 0.05% phosphoric acid solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was set at 260 nm, and column temperature was 30 ℃. The sample size was 10 μL. Using wogonoside as reference,HPLC fingerprints of 10 batches of Qinlian runfei decoction were drawn and the similarity evaluation was conducted with Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition),the common peaks were also confirmed ;the contents of 11 components in Qinlian runfei decoction were determined by the same method. SPSS 21.0 software was used for clus ter analysis ,and SIMCA 14.0 software was used for OPLS-DA to screen marker components affecting quality. RESULTS :There were 21 common peaks in 10 batches of Qinlian runfei decoction ,and the similarity with control fingerprint was greater than 0.98. A total of 11 common peaks were identified , which were rutin , forsythiaside A , forsythiaside B , iris, irigenin, baicalin, forsythiaside, wogonoside, baicalein, irisflorentin and wogonin. The line ar ranges of 11 components were 9.960 0-49.800 0,1.974 0-9.870 0,0.672 0-3.360 0,0.960 0-4.800 0,0.549 0- 2.745 0,5.040 0-25.200 0,1.374 0-6.870 0,0.615 0-3.075 0,0.759 9-3.795 0,0.162 0-0.810 0,0.042 0-0.210 0 μg(all r> 0.999); RSDs of precision , stability (48 h) and repeatability tests were less than 2% ; the average recoveries were 95.81%-100.29% with RSDs of 0.43%-1.73%(n=6);the contents were 8.924 4-12.820 8,0.352 2-0.868 7,0.435 6-0.711 2, 0.389 8-1.309 0,0.335 8-0.530 1,1.680 5-4.542 3,0.701 8-1.584 2,2.240 2-5.442 5,2.351 0-5.558 9,0.106 0-0.182 2,0.076 8- 0.128 9 mg/g,respectively. The results of cluster analysis showed that when class spacing was 10,it could be divided into two groups,S1-S3 and S 4-S10;when the class spacing was 5,the second class could be divided into two categories ,S6,S7,S9 were clustered into one category ,and S 4,S5,S8,S10 were clustered into one category. The results of OPLS-DA analysis showed that S6,S7 and S 9 were at the top of the figure ,S4,S5,S8 and S 10 were at the lower left side of the figure ,and S 1-S3 were at the lower right side of the figure ,which was consistent with the cluster analysis results ;VIP values of baicalin ,iris,forsythiaside A , baicalein and wogonoside were all greater than 1. CONCLUSIONS :Established fingerprint and content determination methods have high precision and good stability. Combined with multivariate statistical analysis ,it can be used for the quality control of Qinlian runfei decoction. Five components as baicalin are the marker components affecting the quality of Qinlian runfei decoction.
5.Population’s acceptance and attitude toward a novel fecal immunochemical test for colorectal cancer screening: a multi-center survey in China
Hong WANG ; Chengcheng LIU ; Fangzhou BAI ; Juan ZHU ; Xinxin YAN ; Mengdi CAO ; Lingbin DU ; Donghua WEI ; Debin WANG ; Xianzhen LIAO ; Dong DONG ; Yi GAO ; Pei DONG ; Chen ZHU ; Yanling MA ; Jing CHAI ; Haifan XIAO ; Yunxin KONG ; Qiang ZHANG ; Weifang ZHENG ; Rongbiao YING ; Hai ZHOU ; Jiansong REN ; Ni LI ; Hongda CHEN ; Jufang SHI ; Min DAI
Chinese Journal of Preventive Medicine 2020;54(7):760-767
Objective:To investigate the acceptance and attitude toward a novel fecal immunochemical test (FIT) in colorectal cancer screening among populations in China.Methods:From May 2018 to May 2019, 2 474 people aged 50-74 years were recruited from five provinces of China (Zhejiang, Anhui, Jiangsu, Hunan and Yunnan). The general demographic characteristics, acceptance of the new FIT technology and operational difficulties through the whole screening process were obtained through questionnaire survey. Multivariate logistic regression model was used to analyze the factors related to difficulties encountered in sampling stool, reading and uploading results.Results:The subjects were (60.0±6.4) years old, and female, high school of above educated, unemployed/retired/other, married and with medical insurance status of “new rural cooperative medical care (NRCMC)” accounted for 61.7% (1 526), 29.0%(718), 34.3% (849), 92.7% (2 293) and 31.3%(775), respectively. The population's acceptance of the FIT technology was 94.8%. In the process of FIT screening, the percentage of occurred difficulties in sampling stool, reading and uploading results were 33.1% (819), 46.4% (1 147) and 62.9% (1 557), respectively. The main difficulties were the uncertainty about whether the sampling operation was standard (28.0%), the inability to accurately judge the result displayed (32.5%) and the need for help without using a smartphone (44.2%). The results of multivariate logistic regression model analysis showed that people aged 65-74 years old and with medical insurance status of “NRCMC” were more likely to encounter difficulties in sampling, and those who were unemployed/retired/other and living with 3 or more family members were less likely to encounter difficulties in sampling. Those aged 65-74 years old, farmers or migrant workers, and those with “NRCMC” were more likely to encounter difficulties in readingresults, and those with 3 or more family members were less likely to encounter difficulties in reading result. Those with “NRCMC” were more likely to encounter difficulties in uploading results, and those with education level of high school or above, living with more than 3 family members were less likely to encounter difficulties in uploading results.Conclusion:The acceptance of the new FIT technology is relatively high among the subjects. Age, education level, occupation, number of family members living together and medical insurance status might be related to difficulties encountered in sampling stool, reading and uploading results, and it can be further strengthened in terms of the technology and characteristics of sub-populations.
6.Population’s acceptance and attitude toward a novel fecal immunochemical test for colorectal cancer screening: a multi-center survey in China
Hong WANG ; Chengcheng LIU ; Fangzhou BAI ; Juan ZHU ; Xinxin YAN ; Mengdi CAO ; Lingbin DU ; Donghua WEI ; Debin WANG ; Xianzhen LIAO ; Dong DONG ; Yi GAO ; Pei DONG ; Chen ZHU ; Yanling MA ; Jing CHAI ; Haifan XIAO ; Yunxin KONG ; Qiang ZHANG ; Weifang ZHENG ; Rongbiao YING ; Hai ZHOU ; Jiansong REN ; Ni LI ; Hongda CHEN ; Jufang SHI ; Min DAI
Chinese Journal of Preventive Medicine 2020;54(7):760-767
Objective:To investigate the acceptance and attitude toward a novel fecal immunochemical test (FIT) in colorectal cancer screening among populations in China.Methods:From May 2018 to May 2019, 2 474 people aged 50-74 years were recruited from five provinces of China (Zhejiang, Anhui, Jiangsu, Hunan and Yunnan). The general demographic characteristics, acceptance of the new FIT technology and operational difficulties through the whole screening process were obtained through questionnaire survey. Multivariate logistic regression model was used to analyze the factors related to difficulties encountered in sampling stool, reading and uploading results.Results:The subjects were (60.0±6.4) years old, and female, high school of above educated, unemployed/retired/other, married and with medical insurance status of “new rural cooperative medical care (NRCMC)” accounted for 61.7% (1 526), 29.0%(718), 34.3% (849), 92.7% (2 293) and 31.3%(775), respectively. The population's acceptance of the FIT technology was 94.8%. In the process of FIT screening, the percentage of occurred difficulties in sampling stool, reading and uploading results were 33.1% (819), 46.4% (1 147) and 62.9% (1 557), respectively. The main difficulties were the uncertainty about whether the sampling operation was standard (28.0%), the inability to accurately judge the result displayed (32.5%) and the need for help without using a smartphone (44.2%). The results of multivariate logistic regression model analysis showed that people aged 65-74 years old and with medical insurance status of “NRCMC” were more likely to encounter difficulties in sampling, and those who were unemployed/retired/other and living with 3 or more family members were less likely to encounter difficulties in sampling. Those aged 65-74 years old, farmers or migrant workers, and those with “NRCMC” were more likely to encounter difficulties in readingresults, and those with 3 or more family members were less likely to encounter difficulties in reading result. Those with “NRCMC” were more likely to encounter difficulties in uploading results, and those with education level of high school or above, living with more than 3 family members were less likely to encounter difficulties in uploading results.Conclusion:The acceptance of the new FIT technology is relatively high among the subjects. Age, education level, occupation, number of family members living together and medical insurance status might be related to difficulties encountered in sampling stool, reading and uploading results, and it can be further strengthened in terms of the technology and characteristics of sub-populations.
7. Effectiveness analysis of intensive management of dyslipidemia in community based on family doctor contracting service
Chengcheng MA ; Meng MENG ; Xiaoqin YE ; Zhizhi JIANG ; Wenyun DAI ; Ying XI
Chinese Journal of Health Management 2019;13(6):490-497
Objective:
To analysis of the effect of strengthening management of dyslipidemia in community based on family doctor contracting service.
Methods:
In December 2014, 1267 cases patients with dyslipidemia aged over 18 years were selected from three community health service centers in Hangzhou, including 645 in intervention group (311 males and 334 females) and 622 in control group (307 males and 315 females). In the intervention group, the management of dyslipidemia in community was strengthened by graded follow-up and personalized diagnosis and treatment based on the contracted services of family doctors, while the control group adopted the general management of dyslipidemia in the community. After 12 months of intervention, the changes of lifestyle (smoking, drinking, body mass index, waist circumference), regularly taking lipid-regulating drugs, blood lipid, blood pressure, blood sugar levels and their control rates were compared by χ2 test or t test before and after intervention between the intervention group and the control group. Non-conditional logistic regression analysis was used to analyze the influencing factors of blood lipid attainment.
Results:
Before intervention, there were no significant differences in gender, age, cardiovascular risk stratification, the levels of lipid and other metabolic indicators, lipid compliance rate between intervention group and control group (
8.miR-1180-5p inhibits proliferation, migration and invasion of prostate cancer cells by activating CDKN1Agene expression
WANG Yong ; GUO Yonglian ; CHEN Lin ; LI Guohao ; Ying Chengcheng ; CHENG Wei
Chinese Journal of Cancer Biotherapy 2018;25(7):698-703
Objective: To study the effects of microRNA-1180-5p (miR-1180-5p) on malignant biological behaviors of prostate cancer VCAP and LNCaP cells and the possible mechanisms. Methods: dsControl (dsControl group) and miR-1180-5p (miR-1180-5p group) were constructed and then transfected into two prostate cancer cell lines VCAPand LNCaP. qPCR and Western blotting were used to analyze the changes in mRNA and protein expressions of CDKN1A, Cyclin D1 and CDK6 after transfection. Cell cycle distribution, proliferation activity, clone formation capacity, cell migration and invasion ability were detected by flow cytometry, MTT assay, colony culture assay and Transwell assay, respectively. Results: qPCR results showed that compared with dsControl, CDKN1A mRNA levels in VCAP and LNCaP cells transfected with miR-1180-5p were up-regulated significantly, while the mRNA expressions of Cyclin D1 and CDK6 were significantly down-regulated (all P<0.01). Western blotting result was consistent with that of qPCR. The percentage of cells in G0/G1 phase was increased after transfection of miR-1180-5p (P<0.05), but the proportion of cells in S phase and G2/M phase was decreased and the cell cycle was arrested at G0/G1 phase (P<0.05). The proliferation activity of the two prostate cancer cells was significantly lower than that of the dsControl group after miR-1180-5p transfection (P<0.05), and the number of colonies in the miR-1180-5p group was significantly lower than that in the dsControl group (P<0.01). In the meanwhile, the cell migration and invasion ability in miR-1180-5p group was decreased (P<0.01). Conclusion: miR-1180-5p can significantly activate CDKN1A gene expression in prostate cancer cells and further inhibit the proliferation, migration and invasion of prostate cancer cells.
9.Pilot study on effect of preventing skin tears with prevention protocol based on guideline
Qixia JIANG ; Xiaoqin HAN ; Ying LI ; Jihong ZHONG ; Yahong LIU ; Juan LI ; Chengcheng FU ; Lili GAO ; Jingbo HUANG ; Xiufang YANG ; Aomei LI
Chinese Journal of Modern Nursing 2016;22(24):3419-3422
Objective To explore the effect of preventing skin tears with prevention protocol based on guideline. Methods According to International Guideline of Prevention Skin Tears to discuss and make prevention protocol, including prediction risk people, prevention fall and fall from bed with auxiliary means, wearing long sleeves clothes, using emollient on skin twice a day, providing nutrients and water, prevention friction and shear force with movement skills, totally six comprehensive measures. Wound care team trained backbone nurses the measures and backbone nurses trained clinical nurses. Before and after intervention 5 month and 17 month, every patient with age more than 18 years and hospitalized more than 24 hours was checked skin from head to toes in order to observe and compare skin tears incidence and practicable rate of prevention measures by cross-sectional survey.Results Before intervention, skin tears incidence was 0.77%(12/1 558), and skin tears incidence after intervention 5 month and 17 month was 0.16%(3/1 777) and 0.22%(4/1 825) ( P<0.05) . Practicable rate of 6 measures in patients with risk and with skin tears after intervention 5 month and 17 month was increased than that before intervention which was unsatisfactory (P>0.05).Conclusions Preven-tion skin tears protocol can reduce skin tears incidence, but practicable rate need be improved. In future nurses training, guidance and management bedside should be strengthened in order to enhance practicable rate of prevention measures.
10.Notch Signaling Pathway with the Polarization of Macrophages
Hongrong LI ; Ying SUN ; Chengcheng CHANG ; Zhenhua JIA
Journal of Medical Postgraduates 2015;(12):1316-1321
Macrophages play an important role in the pathogenesis of many diseases because of its plasticity and diversity.The Notch signaling pathway is a key regulator of the biological function of macrophage and has a complex network connection with many other signaling pathways.This paper reviews the conduction of Notch signaling pathway and its regulation on the polarization of macro-phages.

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