1.Effects of leukemia inhibitor factors on the proliferation and differentiation of cultured human neural stem cells(hNSC) derived from the fetal brains
Guocai YIN ; Zuo LUAN ; Fengqing YAN ; Suqing QU ; Wanli GUO
Chinese Journal of Immunology 1985;0(06):-
0.05) . No significant differences were observed between the expansion folds of hNSC derived from various fetal brain samples when cultured under the same conditions. LIF played great roles on cell proliferation,in LIF + groups, hNSC cell number increased ranging from 4 000-8 400 folds, no cell differentiation occurred; and in LIF" groups,only 43 to 96 folds.The differentiation phenomenons were watched when cultured more than two months. In the course of cell culturing, observed that the effects of LIF on hNSC expansion were obviously demonstrated 50-60 days after inoculation.The number of neurons and astrocytes differentiated from the cultured hNSC were respectively identified by means of Immuno-cytochemical fluorescent assay, and the percentages of neurons(as a proportion of neuron and astrocyte number) were calculated,which were ranging from 12% to 83% in LIF+ cultures, significantly higher than 8% to 23% in LIF- ones(P
2.Analysis of the situation in application of patents associated with plaque researches
Yan XU ; Panpan HU ; Fengqing QI ; Meng ZHANG ; Xiaofang XUE
Chinese Journal of Medical Library and Information Science 2015;24(12):49-53
In order to show the development situation and trend in plaque research, Thomson Innovation Platform-covered application of patents associated with plaque researches was quantitatively analyzed using the Thomson Data Analyzer and Thomson Innovation or other tools, which revealed the overall development situation, the main accepted countries, the main application institutions and the technological direction layout of patents associated with plaque researches.
3.B7H3 upregulates neuron-specific enolase and S100b mRNA expression in mice with S .Pneumoniae meningitis
Yanping WANG ; Xuqin CHEN ; Zhedong WANG ; Bingbing ZHANG ; Fengqing FU ; Yan LI
Chongqing Medicine 2014;(19):2437-2439,2442
Objective To investigate the effect of B7 Homology 3(B7H3)on brain damage of S .Pneumococcal(SP)meningitis . Methods SP meningitis was established by intracerebral ventricular injection of SP suspension on wild-type BALB/C mice .48 mice were divided into 4 groups and received following injections :NS(CON group) ,recombinant murine B7H3alone(B7H3 group) ,SP group ,SP+B7H3 group .At 18 ,48 ,72 h post infection ,mice were conducted neurobehavior score ,then they were anesthetized and killed by cervical vertebra dislocation ,brains were collected .The mRNA expressions of NSE and S100b were detected by real-time PCR .Results Compared with CON group ,the scores of recombinant murine B7H3 group had no significant change at 18 ,48 ,72 h after infection of SP(P>0 .05);at different time points the scores of SP group were decreased significantly than the CON group (P<0 .05);scores of SP+B7H3 group decreased furtherly than SP group(P<0 .05) .The relative expressions of NSE ,S100b mR-NA in brain tissue homogenate :for the NSE ,S100b mRNA relative expressions ,there was no significant difference between B7H3 and CON group at 18 ,48 ,72 h post SP injection(P>0 .05) .At 18h ,48h ,72h ,post infection mRNA expressions of NSE ,S100b in SP group increased compared with CON group(P<0 .05);the mRNA expressions of NSE ,S100b increased furtherly in SP+B7H3 group compared with SP group(P<0 .05) .Conclusion B7H3 upregulates the mRNA expressions of NSE and S100b ,and promotes the progress of SP meningitis in mice .
4.Cloning and expression analysis of the expansin gene RgEXPA10 in Rehmannia glutinosa.
Fengqing WANG ; Yan ZHOU ; Yong HUANG ; Mingjie LI ; Yunhe TIAN ; Fajie FENG ; Xinjian CHEN ; Zhongyi ZHANG
Acta Pharmaceutica Sinica 2015;50(2):233-40
Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.
5.Down-regulation of miR-221-3p/222-3p inhibits cell proliferation and promotes chondrogenic differentiation of human bone marrow mesenchymal stem cells
Jihong YAN ; Shu YANG ; Haimei SUN ; Dandan CAO ; Xiuying ZHANG ; Fengqing JI ; Duo GUO ; Bo WU ; Tingyi SUN ; Deshan ZHOU
Chinese Journal of Tissue Engineering Research 2015;(50):8056-8061
BACKGROUND:The use of mesenchymal stem cels in the field of tissue engineering for osteoarticular injury repair is a very promising tool since these cels are readily expandable and able to differentiate into chondrocytes. Abundant evidence suggests that microRNAs play critical roles in chondrogenic differentiation of mesenchymal stem cels.
OBJECTIVE:To observe the chondrogenic effect of human bone marrow mesenchymal stem cels transfected with lentiviral vectors bearing miR-221-3p/222-3p inhibition, thereby provding new strategies for cartilage injury.
METHODS: miRNA microarray technology was applied to detect microRNAs expression profiles at three different stages of chondrogenic differentiation induction after transforming growth factor-β3 treatment and verified by real-time fluorescence quantitative PCR (RT-qPCR). Human bone marrow mesenchymal stem cels were infected with lentivirus bearing miR-221-3p/222-3p inhibition. After co-suppressing the expression of miR-221/222-3p, cel counting kit-8 was used to determine the cel proliferation, the differentiation of bone marrow mesenchymal stem cels towards chondrocytes was verified by type II colagen protein expression through immunohistochemistry and glycosaminoglycan accumulation was also elevated by sarranine O staining. RT-PCR was used to detect type II colagen and aggrecan mRNA expression at 21 days of chondrogenic induction.
RESULTS AND CONCLUSION: The expression of miR-221-3p/222-3p was inhibited after Lv-miR221-3p/222-3p inhibition co-transfected into bone marrow mesenchymal stem cels. microRNA microarray and RT-qPCR results showed that the expression of miR-221-3p/222-3p was declined significantly at the anaphase of chondrogenic differentiation. The expression levels of chondrogenic markers, Aggrecan and type II colagen were significantly increased in the miR-221-3p/222-3p inhibition group and cel proliferation was also inhibited significantly compared with non-transduced cels or transduced with the empty lentiviral vector group. miR-221-3p/222-3p knockdown in bone marrow mesenchymal stem cels could inhibit proliferation but promote chondrogenic differentiation of bone marrow mesenchymal stem cels.
6.The tentative application of mutual network system in the practice teaching of stomatology
Congchong YANG ; Jin LI ; Fengqing CHU ; Jianping LUO ; Dong HUA ; Bin YAN
Chinese Journal of Medical Education Research 2018;17(4):336-340
The real-time training and mutual network system of stomatology is particularly suited for evaluating the clinical practice training process.This mutual system introduced the closed loop type mutual mode,which ensured the reliability of the evaluation in the practice teaching process by the mutual feedback between students and students,students and teachers.The system solved the deficiency of the previous open loop mutual systemand made the evaluation results more objective.The system realized the process control in practice teaching,and made progress in many ways such as improving the evaluation system of students' abilities,and promoting independent learning and feedback of the teaching.Through this system platform,the students make homework as a link to discover knowledge and further solve the problem.This system guides students to develop the abilities of independent learning and mutual learning,which combines students' mutual evaluation and teachers' evaluation,and shows teaching points comprehensively.The realtime training and mutual network system can explore an effective mean in the transformation education of stomatology,and further meet the new requirements of the social development.
7.Assay development for determination of DZ2002, a new reversible SAHH inhibitor, and its acid metabolite DZA in blood and application to rat pharmacokinetic study
Weiwei JIA ; Jing LI ; Feifei DU ; Yan SUN ; Fang XU ; Fengqing WANG ; Olajide-E. OLALEYE ; Danghui CHEN ; Wei TANG ; Jianping ZUO ; Chuan LI
Journal of Pharmaceutical Analysis 2019;9(1):25-33
Methyl (S)-4-(6-amino-9H-purin-9-yl)-2-hydroxybutanoate (DZ2002) is a potent reversible inhibitor of S-adenosyl-L-homocysteine hydrolase (SAHH). Due to its ester structure, DZ2002 is rapidly hydrolyzed in rat blood to 4-(6-amino-9H-purin-9-yl)-2-hydroxybutyric acid (DZA) during and after blood sampling from rats; this hampers accurate determination of the circulating DZ2002 and its acid metabolite DZA in rats. To this end, a method for determining the blood concentrations of DZ2002 and DZA in rats was developed by using methanol to immediately deactivate blood carboxylesterases during sampling. The newly developed bioanalytical assay possessed favorable accuracy and precision with lower limit of quantification of 31 nM for DZ2002 and DZA. This validated assay was applied to a rat pharmacokinetic study of DZ2002. After oral administration, DZ2002 was found to be extensively converted into DZA. The level of systemic exposure to DZ2002 was significantly lower than that of DZA. The apparent oral bioavailability of DZ2002 was 90%–159%. The mean terminal half-lives of DZ2002 and DZA were 0.3–0.9 and 1.3–5.1 h, respectively. The sample preparation method illustrated here may be adopted for de-termination of other circulating ester drugs and their acid metabolites in rodents.
8.The predictive value of HEART, TIMI and GRACE scores in patients with non-ST-segment elevation myocardial infarction
Yao YU ; Dongxu CHEN ; Fengqing LIAO ; Xiangpeng ZENG ; Yan YANG ; Siying ZHOU ; Wanqing MU ; Yannan ZHOU ; Guorong GU ; Zhenju SONG ; Chenling YAO ; Chaoyang TONG
Chinese Journal of Emergency Medicine 2020;29(7):908-913
Objective:To compare the predictive value of the HEART, TIMI and GRACE scores for major adversecardiovascular events (MACEs) at 7 and 28 days in patients with actue non-ST-segment elevation myocardial infarction (NSTEMI).Methods:More than 12 000 patients with chest pain from the Emergency Department of Zhongshan Hospital Affiliated to Fudan University from October 2017 to October 2018 were studied, including 566 patients with cardiogenic chest pain, 105 patients with ST-segment elevation myocardial infarction (STEMI) excluded and 15 patients lost to follow-up. Finally, 109 patients with NSTEMI and 337 non-myocardial patients with cardiogenic chest pain were enrolled. NSTEMI patients were divided into subgroups according to whether MACEs occurred. LSD t-test, Mann-Whitney U test or χ2 test were used to analyze and compare the differences between the two subgroups about the baseline data, clinical data, HEART, TIMI and GRACE scores at the time of visit. Multivariate logistic regression analysis was used to explore the independent factors of MACEs at 7 and 28 days. And the predictive values of different scores for 7-day MACEs and 28-day MACEs were compared in NSTEMI patients through the receiver operating characteristic (ROC) curve. Results:Compared NSTEMI patients with non-myocardial patients with cardiogenic chest pain, we found a statistically significant differences in sex, past history of coronary heart disease,≥3 risk factors for atherosclerosis, electrocardiogram, high-sensitivity troponin T (hs-cTnT), creatinine value, past history of myocardial infarction, HEART score, TIMI score and GRACE score. In further subgroup analysis of NSTEMI patients who were divided according to whether MACEs occurred, we found previous history of stroke and increased hs-cTnT were statistically different in 7 days after the onset of the disease. The multivariate analysis showed that the previous history of stroke and increased hs-cTnT were independent factors for the occurrence of MACEs at 7 days after the onset of NSTEMI; The previous history of stroke and increased hs-cTnT, electrocardiogram ST segment depression and TIMI score were statistically different at 28 days after the onset of NSTEMI. The multivariate analysis showed that the previous history of stroke and TIMI score were independent factors for the occurrence of MACEs at 28 days after the onset of NSTEMI patients. ROC curve indicated that the predictive value of TIMI score (AUC=0.715, 95% CI: 0.482-0.948) was better than HEART (AUC=0.659, 95% CI: 0.414-0.904) and GRACE scores (AUC=0.587, 95% CI: 0.341-0.833)in predicting MACEs in NSTEMI patients. Conclusions:HEART score, TIMI score and GRACE score can be used to evaluate NSTEMI patients. There is an independent predictive value on TIMI score for the occurrence of 28-day MACEs in NSTEMI patients.
9.Discussion on the relationship between the disposal time of hypobaric oxygen chamber and the establishment of rat cardiac arrest model at high altitude
Jie LIU ; Zengwen MA ; Xiaoxu SHI ; Yan WU ; Cuoji NAN ; Fengqing SONG ; Bin ZHANG
Chinese Critical Care Medicine 2024;36(1):82-85
Objective:To establish the rat cardiac arrest model in high-altitude hypobaric hypoxia environment, and to explore the effect of the treatment time in the hypobaric oxygen chamber on the reproduction of high-altitude rat cardiac arrest model.Methods:SPF grade healthy male Sprague-Dawley (SD) rats were used as observation subjects. The experiment was conducted in two different altitude areas. The rats from the Plateau Branch of Institute of Cardiopulmonary and Cerebral Resuscitation of Sun Yat-sen University (Xining, Qinghai) were weighed and numbered, and they were placed in a hypobaric oxygen chamber (simulated altitude of 3 000 meters, speed of ascent and descent of 15 m/min, temperature of 20 ℃, cabin pressure of 69.5 kPa, cabin oxygen pressure of 14.5 kPa). After 30 days of feeding, the rats were obtained according to random number table method, and the cardiac arrest model was established by asphyxia method as the 30-day hypobaric hypoxia group. After 60 days of feeding, rats were randomly selected again, and the cardiac arrest model was established as the 60-day hypobaric hypoxia group. Thirty rats were randomly selected from the Institute of Cardiopulmonary Cerebral Resuscitation at Sun Yat-sen University (Guangzhou, Guangdong) by the same method, and the cardiac arrest model was established as the plain control group. The differences in the body weight of rat modeling precursors and the induction time of asphyxia during the modeling process among different groups were compared.Results:Finally, cardiac arrest model was established in 16 rats in the 30-day hypobaric hypoxia group and in 22 rats in the 60-day hypobaric hypoxia group. There was no significant difference in the body weight of rats before modeling among the plain control group, 30-day hypobaric hypoxia group and 60-day hypobaric hypoxia group [g: 429.00 (389.25, 440.75), 440.00 (415.50, 486.25), 440.00 (400.00, 452.50), all P > 0.05]. The asphyxia induction time of rats in the 60-day hypobaric hypoxia group was significantly longer than that in the 30-day hypobaric hypoxia group (s: 294.59±75.39 vs. 234.31±93.86, P < 0.01), even about 1.4 times of the plain control group (s: 294.59±75.39 vs. 208.73±30.88, P < 0.01). There was no significant difference in the asphyxia induction time between the 30-day hypobaric hypoxia group and the plain control group ( P > 0.05). Conclusion:Rats treated in a hypobaric oxygen chamber for 60 days are more suitable for the preparation of high-altitude cardiac arrest model, and are also consistent with the oxygen reserve and hypoxia tolerance of high-altitude rats.