1.Identification of daturae flos and its adulterants based on DNA barcoding technique.
Jianping HAN ; Meini LI ; Kun LUO ; Meizi LIU ; Xiaochen CHEN ; Shilin CHEN
Acta Pharmaceutica Sinica 2011;46(11):1408-12
To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnH, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.
2.Effect and mechanism of heat shock protein inhibitors on cell proliferation and apoptosis in human nasopharyngeal carcinoma
Qiuxia PANG ; Jingzi HE ; Jumei ZHAO ; Jinghong XU ; Aihong WANG ; Meini CHEN ; Tao LIU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2017;24(8):385-389
OBJECTIVE To study the effect of different concentrations of 17-AAG and EGCG monotherapy or in combination on the induced apoptosis in human nasopharyngeal carcinoma, and to explore new molecular targets for the treatment of nasopharyngeal carcinoma. METHODS MTT colorimetric method and fluorescent staining were used to detect the change of CNE proliferation inhibition rate and cell morphology. And furthermore, the expression level of Bcl-2, Bax, Caspase-3 were detected by RT-PCR. RESULTS 1. 17-AAG or EGCG alone had inhibitory effect on the human nasopharyngeal carcinoma CNE cells at 24 h, 48h and 72 h, and it was related with time and dose(P<0.01). The inhibition effect of combination of 17-AAG and EGCG was significantly increased,which was time and dose dependent(P<0.01). 2. RT-PCR was used to detect the mRNA expression level of Bcl-2, Bax and caspase-3. The level of Caspase-3 and Bax mRNA expression after treated by 17-AAG and EGCG was significantly higher, and the level of bcl-2 mRNA expression was lower than that after treated by 17-AAG or EGCG alone. CONCLUSION Our investigation implied that 17-AAG and EGCG in combination can effectively inhibit the proliferation of human nasopharyngeal carcinoma CNE cells. The involved mechanisms may be associated with the upregulation of Bax and Caspase-3 expression.
3.Effects of HSP90 inhibitor 17-AAG on cell cycle and apoptosis of human gastric cancer cell lines SGC-7901.
Meini CHEN ; Jinghong XU ; Jumei ZHAO
Journal of Southern Medical University 2013;33(2):271-275
OBJECTIVETo study the effect of the HSP90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on cell proliferation and apoptosis of human cancer SGC-7901 cells and explore the mechanisms.
METHODSThe inhibitory effect of 17-AAG on the proliferation and morphology of SGC-7901 cells was assessed with MTT assay and DNA-PI staining, respectively. Flow cytometry was employed to analyze the changes in cell cycle and apoptosis of the cells following 17-AAG exposure. The cellular expression of Fas protein was detected by immunohistochemistry.
RESULTS17-AAG significantly suppressed the proliferation of SGC-7901 cells in a time- and dose-dependent manner. After treatment with 17-AAG for 48 h, SGC-7901 cells showed cell cycle arrested at G(2)/M stage, and the cell apoptosis rate increased with the 17-AAG concentration. The expression of Fas protein in the cytoplasm of SGC-7901 cells increased gradually with the increase of 17-AAG concentration.
CONCLUSION17-AAG can induce apoptosis, alters the cell cycle distribution and up-regulates the expression of Fas protein in SGC-7901 cells to suppress the cell proliferation.
Apoptosis ; drug effects ; Benzoquinones ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; pharmacology ; Stomach Neoplasms ; pathology ; fas Receptor ; metabolism
4.Design and effect evaluation of high-fidelity simulation teaching in emergency and critical care nursing
Jianling SUN ; Liming SUN ; Meini ZHANG ; Haihua ZHU ; Weigeng SHEN ; Haibin CHEN ; Xiaowen PAN ; Yang LIN
Chinese Journal of Modern Nursing 2020;26(33):4697-4701
Objective:To explore the teaching effect of high-fidelity simulation teaching in emergency and critical care nursing course.Methods:From March to June 2019, 56 four-year nursing undergraduates from Xiamen Medical College in 2016 were selected as research subjects by convenient sampling. The high-fidelity simulation teaching was carried out in the emergency and critical care nursing course. The Jeffries Simulation Design Scale (SDS) and open-group interviews were used to evaluate the learning effect.Results:Among 56 students, scores of simulation teaching confidence and satisfaction were (3.80±0.80) and (4.00±0.62) respectively. Group interviews showed that students directly applied the theoretical knowledge they had learned to work situations, and students enhanced their self-learning ability and reflected on operational errors in simulation so as to provoke thinking and enhance learning confidence and personal learning ability.Conclusions:The application of high-fidelity simulation teaching in emergency and critical care nursing course can improve the self-confidence, teamwork ability and comprehensive quality of nursing undergraduates, and increase teaching satisfaction, which is feasible.
5.Prognostic value of thromboelastography maximum amplitude and arterial blood lactate levels for sepsis in elderly patients
Jifang LIANG ; Xiuzhe WANG ; Xiaojing YANG ; Weidong WU ; Wenjing WU ; Meini JIANG ; Shuxian CHEN
Chinese Journal of Geriatrics 2022;41(2):168-172
Objective:To evaluate the prognostic value of thromboelastography maximum amplitude(MA)and arterial blood lactate levels for sepsis in elderly patients.Methods:A retrospective analysis was performed on clinical data of 63 sepsis patients(≥60 years old)admitted to the Intensive Care Unit(ICU)of Bethune Hospital of Shanxi Province from December 2018 to February 2020.MA values, white blood cell counts, lymphocyte counts, platelets, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)scores, sequential organ failure assessment(SOFA)scores, underlying diseases, body mass index, laboratory test results and other related treatments were analyzed.The subjects were divided into the survival group and the death group according to the 28-day survival outcome.Differences in MA, APACHE Ⅱ scores, SOFA scores and laboratory test results between the two groups were analyzed, and the correlations of MA with infection parameters and age were examined.Influencing factors of survival outcomes were analyzed using multivariate Logistic regression.The receiver operating characteristic curve(ROC)was used to calculate the prognostic value of MA and arterial lactate for sepsis in elderly patients.Results:The main sources of infections were pulmonary and abdominal(79.4%, 50/63)in 63 elderly patients with sepsis.The incidences of positive blood cultures and deaths were 15.9%(10/63)and 66.7%(42/63), respectively.There existed significant differences in lymphocyte counts, arterial lactate levels, MA and lengths of stay in the ICU between the survival group and the death group( t=3.847, 2.153, 2.745, -3.574, respectively, all P<0.05).MA was correlated with arterial lactate, SOFA score and survival outcome( r=-0.498, -0.506, and -0.358, respectively, all P<0.05).Multivariate Logistic regression analysis showed that MA and arterial lactate were independent factors for the survival outcome( OR=1.626, 0.766, all P<0.05).The area under the ROC curve(AUC, 95% CI)for the combination of MA and arterial lactate was larger than that of either MA or arterial lactate alone(0.89, range: 0.763-0.846; 0.58, range: 0.574-0.730; 0.77, range: 0.521-0.832; all P<0.05). Conclusions:The combination of thromboelastography maximum amplitude and lactate in arterial blood has important clinical value in assessing the prognosis of elderly patients with sepsis.
6.Synergistic sensitization of hUMSCs-derived supernatant combined with temozolomide in different glioma cell lines
Yusi LIU ; Mingming WANG ; Yufu ZHANG ; Xiaoyan JIN ; Jing HE ; Haiyan SHI ; Meini CHEN ; Jing ZHANG
Chinese Journal of Pathophysiology 2024;40(3):385-394
AIM:To explore the synergistic sensitization effect of human umbilical cord mesenchymal stem cell culture supernatant(hUMSC-CM)combined with temozolomide(TMZ)on various glioma cell lines,and to elucidate the underlying mechanisms.METHODS:The hUMSC-CM was harvested using two different serum deprivation tech-niques at 24 and 48 h,and was converted into freeze-dried powder,which was then given to rat malignant glioma cell line RG-2,human astrocytoma cell line U251 and human glioblastoma cell line LN-428 at 5 concentrations(0,1,3,6 and 9 g/L).The effectiveness and sensitivity of hUMSC-CM for inhibiting growth of glioma cells at 24,48 and 72 h were as-sessed using CCK-8 assay.Hematoxylin-eosin(HE)staining combined with CCK-8 assay was employed to evaluate the chemotherapy sensitivity of glioma cells after 48 h of treatment with TMZ at 6 concentrations(0,25,50,100,200 and 400 μmol/L).Two concentrations(3 and 9 g/L)of hUMSC-CM and 3 concentrations(50,100 and 200 μmol/L)of TMZ were chosen for concurrent treatment of glioma cells to assess the proliferation and pathological alterations.TUNEL staining was utilized to detect apoptosis.Flow cytometry was utilized to analyze cell cycle modifications.The expression alterations of apoptosis-inducing proteins,cleaved caspase-3,cleaved caspase-8 and cleaved PARP1,as well as autophagy-inducing proteins beclin-1 and LC3,were examined using Western blot to investigate the synergistic sensitization mechanism of hUMSC-CM combined with TMZ in vitro.RESULTS:The susceptibility of glioma cell lines to hUMSC-CM and TMZ varied,with RG-2 showing the highest sensitivity,followed by U251,and then LN-428.The inhibitory effect of hUMSC-CM(3 and 9 g/L)and TMZ(50,100 and 200 μmol/L)combined treatment on glioma cells was significantly greater than that that of single-agent treatments(P<0.05),demonstrating a dose-and concentration-dependent enhancement.Notably,the combination of 9 g/L hUMSC-CM(C9)with 50 μmol/L TMZ(T50)effectively suppressed glioma cell growth.CCK-8 as-say indicated a significant reduction of cell viability in C9+T50 group compared with either C9 or T50 alone(P<0.05).HE staining and TUNEL staining revealed pronounced morphological changes and significant apoptotic features in glioma cells treated with C9+T50.Flow cytometric analysis confirmed that C9+T50 induced cell cycle arrest in glioma cells.Fur-thermore,compared with control group,the levels of cleaved caspase-3,cleaved caspase-8,cleaved PARP1,beclin-1,and LC3-Ⅱ/LC3-Ⅰ were significantly elevated in the C9+T50-treated glioma cells(P<0.01).CONCLUSION:(1)The concomitant administration of hUMSC-CM and TMZ exerts a broad inhibitory effect on glioma cells,with a synergistic sen-sitization observed across different cell lines.(2)The enhancement of glioma cell sensitivity to TMZ by hUMSC-CM may be attributed to the modulation of caspase-8/caspase-3/PARP1 signaling pathway and the induction of both apoptosis and autophagy in glioma cells.
7.Correlation between the Detection of Multiple Cytokine Levels in Neonatal Umbilical Cord Plasma and Early-onset Neonatal Sepsis
Zhengjiang CHANG ; Qian WEI ; Xiaolong LIU ; Zhaoying FU ; Meini CHEN ; Jinwei HE ; Haiyan FAN
Journal of Modern Laboratory Medicine 2024;39(4):150-154
Objective To investigate correlation between the expression level of multiple cytokine levels in neonatal umbilical cord plasma and early-onset sepsis for screening out the cytokines with good diagnostic value for early-onset neonatal sepsis(EONS).Methods Full-term neonates and preterm neonates(Gestational age ≥ 32 weeks)of 310 cases between September 2021 and June 2023 were selected as study subjects.According to clinical signs,laboratory results and blood culture,these subjects were divided into 3 groups:control group without sepsis,EONS blood culture positive group and EONS blood culture negative group.Umbilical cord blood plasma of all subjects was collected within 72 hours after birth.The expression levels of cytokines IL-2,IL-4,IL-6,IL-9,IL-10,IL-21,IFN-γ and TNF-α were determined,and cytokines with high expression levels(high correlation)were screened out.Receiver operating characteristic(ROC)curve was used to analyze the specificity and sensitivity of the selected cytokines in the diagnosis of neonatal early-onset sepsis.Results Among the 8 cytokines mentioned above,the concentrations of IL-6,IL-9 and IL-21 in cord blood plasma of neonatal early-onset sepsis positive blood culture patients(392.6±258.7pg/ml,11.9±7.5pg/ml,29.1±16.8 pg/ml)and negative blood culture patients(353.8±244.5pg/ml,10.4±6.3pg/ml,27.7±19.2pg/ml)were higher than those of the control group(34.9±25.1pg/ml,5.9±4.5pg/ml,10.8±10.1 pg/ml),with significant differences(t=23.961,20.732;15.174,17.824;22.466,21.193,all P<0.01),and the increase of IL-6 concentration was the most obvious.ROC curve analysis(the cut-off values of IL-6,IL-9 and IL-21:123.0 pg/ml,3.60 pg/ml,6.00 pg/ml,respectively)showed that the areas under the ROC curve for IL-6,IL-9 and IL-21 alone detection were 0.876(95%CI:0.786~0.955),0.782(95%CI:0.667~0.875)and 0.825(95%CI:0.737~0.913),respectively.The area under the ROC curve for the combined detection of IL-6,IL-9 and IL-21 was 0.930(95%CI:0.875~0.997).The combined detection of IL-6,IL-9 and IL-21 improved the specificity and sensitivity of the test than IL-6,IL-9 and IL-21 alone detection,and the differences were statistically significant(Z=2.137,2.391,2.257,all P<0.05).There was no significant difference in cytokine expression between positive blood culture and negative blood culture neonates with early-onset sepsis(t=0.276~3.377,all P>0.05).Conclusion The cytokines expression of IL-6,IL-9 and IL-21 in neonatal umbilical cord plasma of neonatal early-onset sepsis were increased.Combined detection of IL-6,IL-9 and IL-21 has good diagnostic value for early-onset neonatal sepsis.
8.Establishment and Preliminary Application of Competence Framework for Clinical Teaching Management Positions Based on Nominal Group Technique
Yizhen WEI ; Duoduo ZHANG ; Qiang WANG ; Xiao CHANG ; Meini CHEN ; Linzhi LUO
Medical Journal of Peking Union Medical College Hospital 2024;15(3):718-723
To develop a framework for clinical teaching management (CTM) position competencies, and to initiate the preliminary application of this framework. Key groups comprising clinical teaching coordinators, clinical trainees, teaching administrators, and the educational reform practice and research group at Peking Union Medical College Hospital (PUMCH) were selected as research samples. The nominal group technique (NGT) was employed to generate, integrate, and establish a framework, as well as to describe CTM competencies. Additionally, rankings of competency importance were gathered. The competency framework was then utilized to conduct a self-assessment of CTM through a questionnaire survey, scored using the Likert method (ranging from 1 to 5 points), followed by an analysis of the results. The initial NGT discussion round among key groups led to a consensus on the primary CTM competency framework, which included professionalism, coordinating ability, teaching ability, and teaching innovation. Divergent views emerged regarding the connotations and importance rankings of each dimension. Following the second NGT discussion by the educational reform practice and research group, a self-assessment was carried out using the CTM competency framework. This second discussion round resulted in a competency framework for clinical teaching management positions that encompassed seven dimensions: medical knowledge and skills, learner-centeredness, communication and cooperation, professionalism and role modeling, reflection and improvement, teaching theory and methods, and teaching coordination and management. The self-assessment revealed that, aside from the dimensions of teaching coordination and management and teaching theory and methods, the scores for the other five dimensions were above 80% (4 points). A preliminary CTM competency framework has been established and applied. The results provide significant insights for guidiny the enhancement and evaluation of competencies.However, specific items within this framework require further validation and refinement through practical application.