Aim To establish an in vitro early drug cardiac tox-icity evaluation method by human embryonic stem cells derived cardiomyocytes ( hESC-CM) and real-time cell analysis Cardio (RTCA Cardio) system. Method The hESC-CM were cultured at RTCA Cardio E-Plate 96. Impedance signals from hESC-CM were analyzed for beating rate, contraction amplitude and beating rhythm irregularity to determine the optimum inoculation density and detection duration. Based on this, we used 0. 1 % DMSO to be the solvent and quinidine (0. 2, 0. 78, 3. 13, 12. 5, 50 and 100 μmol·L - 1 ) known as affecting cardiac activity to validate this method. Result The results revealed no significant changes in the cell index (CI), transient pulse patterns, beating rate and amplitude of hESC-CM. Quinidine will affect the CI and transi-ent pulse patterns of hESC-CM and decrease the beating rate and amplitude of hESC-CM when its concentration ≥3. 13 μmol · L - 1 . And this effect is concentration-dependent, the higher the concentration,the more time they need to recover beating and the more significant the beating rate and amplitude inhibition of quinidine on hESC-CM. Conclusion The method established by hESC-CM and RTCA Cardio system can detect the effect of quinidine on the contraction of hESC-CM, and this indicates that this method has the potential to be an attractive high-throughput tool for screening potential drugs in early evaluation of drug car-diotoxicity.

Objective To discuss the effect of the teaching mode of technological pedagogical and content knowledge in biochemistry theory teaching. Methods 400 students in 8 classes of clinical medicine undergraduate in Grade 2013 were selected and randomly divided into 2 groups. The traditional teaching mode was applied in the control group of 198 students in 1-4 classes, while the technological pedagogical and content knowledge teaching mode was applied in the experimental group of 202 students in 5-8 classes. 385 students in 8 classes of clinical medical undergraduates in Grade 2014 were selected and randomly divided into 2 groups. The control group of 189 students in 1-4 classes adopted the traditional teaching mode, while the experimental group of 196 students in 5-8 classes adopted the technological pedagogical and content knowledge teaching mode, which used the micro-lesson and network platform as learning resource superior to the flipped classroom. Flipped classroom was divided into two major learning links:extracurricular self-study and class digestion. Through the network platform the micro-lesson was presented to the learners. Learners could make self-study according to their own specific circumstances and in the classroom many activities were increased such as the mutual cooperation between the students, the students' PPT teaching, the students' questions and the discussion, and the interaction between teachers and students, etc. The results of the examinations of the two terms students of the same profession and the questionnaire were analyzed. The related data were processed by SPSS 15.0, and the data between groups were compared by t test . Results The test scores analysis showed that the individual test scores in experimental group of Grade 2013 were [(17.94±2.02) vs. (12.28±4.17)], and the individual test scores in experimental group of Grade 2014 were [(18.21 ±1.78) vs. (12.45 ±5.13)], which were obviously higher than the control group, and there was statistical significance. The final exam scores in experimental group of Grade 2013 were [(78.28±11.18) vs. (68.65±12.51)], and the final exam scores in experimental group of Grade 2014 were [(81.73 ±9.12) vs. (74.41 ±11.87)], which were obviously higher than the control group, and there was statistical significance. The results of survey showed that the students thought the teaching mode aroused their study interests while 393 (93.7%), thought the teaching mode developed their self-study ability while 357 (89.7%), thought the teaching mode beneficial to cultivating their ability of solving the problems. Conclusion The teaching mode of technological pedagogical and content knowledge is of certain signifi-cance to break the plight of the traditional teaching, inspire the students interest in learning, improve the teaching quality of biochemistry, and make for the teachers' professional development.

Objective To find the plantar pressure readings which best indicate foot inversion during the stance phase of walking among hemiplegic stroke survivors.Methods Twenty-two hemiplegic stroke survivors who were able to walk without extra aid were recruited as the experimental group, while 17 healthy elderly men of similar age and body weight were selected as the control group.Those in both groups were asked to walk at their preferred speed over a Footscan device which measured medio-lateral pressure ratios, maximum plantar pressures and the contact areas of both feet.The Clinical Spasticity Index (CSI) was used to evaluate the affected feet.Results The average medial forefoot pressure of the affected side in the experimental group was significantly lower than that of the control group, but their average mesopodium and heel pressure was significantly higher.The average pressure applied by the great toe on the uninjured side in the experimental group was significantly smaller than that of the control group, but the average mesopodium and heel pressure of both feet among the hemiplegics were significantly higher than those of the control group.Among the experimental group, the average medial forefoot pressure of the affected foot was significantly greater than that of the healthy foot.The average contact area of the great toe on the affected side was significantly bigger than was observed in the control group.That of the medial forefoot was, however, significantly smaller than in the control group.There was no significant difference in the contact area between the healthy and affected feet in the experimental group, though the maxmium medio-lateral pressure ratios of their full feet and forefeet on the affected side were significantly lower than those in the healthy group.No significant differences in the maxmium medio-lateral pressure ratios of the heel were observed between the two groups, nor of the full feet, forefeet and heels of the affected and unaffected sides in the experimental group.The patients demonstrated consistently reduced joint mobility on both sides during the stance phase, coinciding with increased inversion.A significant negative correlation was found between the maxmium medio-lateral pressure ratios of the full foot and the maximum pressure of the lateral part of forefoot in the experimental group, but there was no significant correlation with contact area or CSI.Conclusions Plantar pressure data can be used to describe the amount of foot inversion in the stance phase of walking with hemiplegic patients after stroke.The maxmium medio-lateral pressure ratios can effectively reflect their foot inversion.

Objective:To observe the long-term efficacy and late adverse reactions of simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT) for mediastinal lymph node recurrence after radical surgery for esophageal squamous cell carcinoma (ESCC).Methods:A total of 20 ESCC patients with mediastinal lymph node recurrence (≤5) after radical surgery admitted to Department of Radiotherapy, Shanghai Ruijin Hospital between June 2019 and December 2021 were enrolled in this prospective study. Among them, 10 patients were enrolled in phase I study and 10 patients in phase II study. Four, 3 and 13 patients received three different doses of SIB-IMRT at 58.8 Gy/28 fractions, 64.4 Gy/28 fractions and 70.0 Gy/28 fractions for recurrent lesions, respectively. The overall survival (OS) rate, local control rate (LCR) and progression-free survival (PFS) were calculated by Kaplan-Meier analysis. Adverse reactions were also analyzed.Results:The most common sites of recurrence were 2R and 4 L, accounting for 35% and 25%, respectively. The median follow-up time was 32 months. For patients who received salvage chemoradiation after relapse, the 1-, 2- and 3-year OS rates were 100%, 88% and 78%, the 1-, 2- and 3-year PFS rates were 85%, 78% and 78%, respectively. The most common hematological toxicities were leukocytopenia and anemia. The most common nonhematological toxicity was esophagitis. However, no grade 3 or above esophagitis, pneumonia and cardiotoxicity were found. Three patients who received SIB-IMRT at 58.8 Gy/28 fractions died of distant metastases at 2 years after treatment, and 1 patient who received SIB-IMRT at 70.0 Gy/28 fractions died of distant metastases at 16 months after treatment.Conclusion:Salvage chemoradiotherapy using SIB-IMRT is efficacious and safe for mediastinal lymph node recurrence in ESCC patients after radical resection.

BACKGROUND:Currently, hematopoietic stem celltransplantation mainly depends on unrelated donors. Mental state of the unrelated donors is very important to ensure the successful celltransplantation. OBJECTIVE:To compare mental and physical health status of relative and unrelated donors during the hematopoietic stem cellcol ection. METHODS:We compared the mental (Symptom Checklist-90) and physical (temperature, breath, pulse, and blood pressure) health status of relative and unrelated donors at admission, 1 day before cellcol ection, and 1-2 days after cellcol ection.RESULTS AND CONCLUSION:At admission, there was no difference in the mental health status of relative and unrelated donors (P>0.05), while the scores on Symptom Checklist-90 were significantly higher in the unrelated donors than the relative donors, including total score, forced, depression, anxiety, hostility, and fear (P<0.05). The physical signs were steady in the unrelated and relative donors, but the difference in breath and systolic blood pressure was of great significance before and after cellcol ection in the two groups. These findings indicate that during cellcol ection, the unrelated donors exhibit heavier mental load than the relative donors, and psychological counseling and health guidance are necessary.

Objective To explore the method and effect of applying just-in-time-teaching (JiTT) mode in the reform of the teaching method of biochemical comprehensive experiment (Isolation, Purification and Identification of γ-Globulin From serum). Methods A total of 100 undergraduate students in our 2016 clinical medicine were divided evenly into two groups: the control group and the experimental group. The control group used traditional teaching, and the experimental group used JiTT-based teaching. In the experimental group, an online environment for pre-classroom discussion and feedback was constructed by uploading learning resources such as short video and tutorials to the online teaching platform and the online learning exchange group 3-5 days before the class; the class session itself consisted of personal tests, face-to-face teaching by teachers, group discussion, finishing experimental report and answering questions etc., and was intended to evaluate the students' self-study. SPSS 22.0 was used to collect exam scores and questionnaire results, and the data between groups were compared using t test. Results The scores of individual test scores in the experimental group (9.44±0.59) were higher than those in the control group (8.77±0.41) (P=0.00), and the difference was statistically significant. In all the six questionnaire items pertaining to teaching effect , JiTT teaching method received higher scores than the traditional teaching method (P<0.05), and the difference was statistically significant. Conclusion JiTT can improve the quality of biochemistry experiment teaching. It is worthwhile to try it in the teaching of other basic specialized courses.

Objective:To investigate humoral and antigen-specific T cell immune responses induced by oral administration of WGP-OVA in C57BL/6 mice.Methods:C57BL/6 mice were orally administered with PBS,WGP and WGP-OVA for 17 days,respec-tively.The serum immunoglobulin levels including IgG1,IgG2a,IgG3 and IgM of mice were detected using LEGENDplexTM Multi-An-alyte Flow Assay Kit.HE staining was used to compare the size of lymphoid nodules in spleens.FACS was used to analyze the percent-age of F4/80+macrophages and F4/80+SIINFEKL+macrophages in inguinal lymph nodes(ILNs).In addition,the T cell-mediated im-munity stimulated by the WGP-OVA oral vaccine was analyzed by ILNs-infiltrating antigen-specific CD8+T cell using MHC Tetramer assay.Results:Oral administration of WGP-OVA significantly increased the contents of IgG1,IgG2a,IgG3 and IgM in the serum,and enlarged the lymphoid nodules in spleens,compared to those orally administered with PBS.Oral administration of WGP-OVA pro-moted the ILNs-infiltration of F4/80+macrophages,and enhanced the antigen-presentation of OVA in macrophages in ILNs.Moreover,the expansion of OVA specific CD8+CTLs in ILNs was also stimulated by WGP-OVA.Conclusion:WGP-OVA involved in both the humoral and antigen-specific T cell immune responses in C57BL/6 mice.

Objective:To investigate the effect of tripartite motif-containing 23 (Trim23) on the differentiation and maturation of dendritic cells and the possible mechanism.Methods:Mouse bone marrow-derived dendritic cells (BMDCs) were prepared from bone marrow cells of C57BL/6 mice with the presence of Flt3L. Real-time quantitative PCR and Western blot were used to detect the expression of Trim23 in BMDCs after LPS stimulation. An overexpression vector for full-length Trim23 (Trim23 OE) was constructed and transfected into BMDCs, and the pcDNA3.1 empty vector was used as control. Flow cytometry was used to detect the expression of CD80, CD86, CD40 and MHCⅡ on the surface of vector-transfected BMDCs after LPS stimulation and ELISA was used to detect the secretion of IL-12p40, TNF-α, IL-6 and IL-10 by these cells. CD8 + and CD4 + T cells were isolated from spleen and lymph nodes of OT-Ⅰ and OT-Ⅱ mice by magnetic beads and co-cultured with LPS-treated BMDCs in the presence of ovalbumin (OVA). Flow cytometry was used to detect the proliferation and differentiation of CD8 + and CD4 + T cells. Western blot was performed to analyze the phosphorylation of p38, ERK1/2 and AKT in BMDCs. Two overexpression vectors for Trim23 mutants lacking RING or ARF domain (Trim23 ΔRING and Trim23 ΔARF) were constructed and transfected into BMDCs. Then flow cytometry and ELISA were used to detect the expression of surface molecules and cytokines. Results:The expression of Trim23 in BMDCs was significantly down-regulated after LPS stimulation. The expression of MHCⅡ, CD86 and CD80 and the secretion of TNF-α and IL-6 decreased significantly in BMDCs overexpressing Trim23. Furthermore, overexpression of Trim23 inhibited the ability of BMDCs to induce the proliferation and differentiation of CD4 + T cells and the proliferation of CD8 + T cells. Western blot showed that the phosphorylation of p38 and ERK1/2 decreased significantly in Trim23-overexpressing BMDCs. Compared with wildtype Trim23, overexpression of Trim23 ΔRING had no significant influence on the expression of surface molecules (MHCⅡ and CD86) and the secretion of cytokines (TNF-α and IL-6) in BMDCs stimulated by LPS. Conclusions:Trim23 overexpression inhibited the maturation and immune activation of BMDCs via MAPK signal pathway and its RING domain. This study provided reference for targeting Trim23 to improve the immune response of dendritic cell-based tumor vaccines.

Objective:To investigate the application effect of the blended teaching model of "pre-class synchronous small private online course (SPOC)+ flipped classroom+post-class knowledge expansion" in the theoretical teaching of biochemistry and molecular biology. Methods:A total of 951 students majoring in clinical medicine, preventive medicine, and pharmacy in the class of 2020 in Qiqihar Medical College were selected as teaching reform research group (experimental group), and their curriculum relied on the self-built massive open online course (MOOC) curriculum and the MOOC resources of Chinese universities to construct a blended teaching model; the reform methods such as small class grouping, case-based learning, and diversified examination were adopted to carry out theoretical flipped teaching activities with cross integration of cases and knowledge points as the main line. A total of 847 students majoring in clinical medicine, preventive medicine, and pharmacy in the class of 2019 were selected as control group, and the traditional teaching method was used for theoretical courses. The courses were evaluated by offline assessment and online assessment, and feedback information was collected through examination scores, questionnaire survey, and online voting. SPSS 20.0 was used for statistical analysis, and the t-test was used for comparison between groups. Results:The experimental group had a significantly higher total score of final examination than the control group [(92.12±3.88) vs. (86.73±5.27), P<0.05]. The questionnaire survey showed that the students in the experimental group showed a relatively high degree of satisfaction with the blended teaching reform, which increased their participation, experiencing, and sharing activities. The students majoring in clinical medicine in the class of 2020 believed that they had established clinical thinking ability (263, 92.61%) and become familiar with the clinical manifestations and pathogenesis of common diseases (262, 92.25%); the students majoring in preventive medicine in the class of 2020 believed that they had increased their awareness of serving the public (151, 93.21%) and developed the ability to teach and guide healthy living (148, 91.36%); the students majoring in pharmacy in the class of 2020 believed that they had mastered the mechanism of action of drugs for disease treatment (138, 93.24%) and understood the importance of rational drug use in clinical practice (135, 91.22%). Conclusion:The blended teaching model realizes student-centered teaching, stimulates the interest and initiative in learning, and improves learning outcome, thereby improving teaching quality to a certain extent.

OBJECTIVE：To study the effects of gin seng root extract on autophagy ，proliferation and cytokine of splenic lymphocyte of mice ，and to study its mechanism. METHODS ：The splenic lymphocyte of mice were divided into blank control group，positive control group （concanavalin A ，5 μg/mL），different concentration of ginseng root extract groups （32，125，500 μg/mL，by lyophilized powder ）. After 48 h of culture with the corresponding medicine ，acridine orange staining method was used to detect autophagy of splenic lymphocyte ；CCK-8 assay was used to detect the cell proliferation ；ELISA assay was used to determine the levels of IL- 4，IL-6 and TNF-α in cell culture；Western blotting method was used to detect the expression of LC 3B and Beclin- 1，as well as the phosphorylation of AMPK ，AKT and mTOR. RESULTS ：Compared with blank control group ，32， 125，500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice（P＜0.05 or P＜0.01）；125，500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes，the levels of IL- 4，IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ，significantly increased the protein expression of Beclin- 1， quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ，2015，23（3）：232-243. the phosphorylation of AMPK protein ，and significantly reduced the phosphorylation level of AKT and mTOR proteins ，with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS ：Ginseng root extract can induce autophagy of mice splenic lymphocytes，activate the activity of splenic lymphocytes ，regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ，so as to exert immune enhancement effect.