Teachers applied task-based learning and Team-based learning methods in the course of ethnic medicine.Teaching contents were classified according to relations of different herbs and quantity of herbs in each chapter.7-9 students formed a learning group which was also called team.Learning groups prepared lessons before class,debated on different tasks such as herb identification and treatment prescription of clinical cases in class,and drew a conclusion after debates.Learning groups expounded their viewpoints and debated with other learning groups about different views.Teachers recorded and commented on viewpoints from each learning group.After class,through a questionnaire,teachers understood the students' feedback to the whole teaching process,and the effect of application of task driven and cooperative team learning got students' recognition.In the future,we will make some improvement by increasing the classroom comment,improving the preparation before class,increasing the efficiency of learning group presentation and debate time,enriching teachers' academic knowledge and improving their teaching skills.

Objective To discuss task-based learning and team-based learning methods and lecture-based learning method in the class of ethnic medicine.Method 50 students in clinic medicine (general practice) of grade 2012 were selected as D-TBL group and 54 students in clinic medicine of grade 2013 were selected as LBL group.Both groups have teaching content,textbook,teachers,class hours in common.Effect of teaching was valued by tests,evaluation in students,questionnaires.SPSS was used to analyze scores of tests.T test was used.Results The correct answer rate?of subject items in D-TBL group was higher than that of LBL group and the difference had statistical significance [(94.56 ± 4.95)％ vs.(29.26 ± 12.15)％,t=36.382,P=0.000).There was no significant difference between the correct answer rate of personal test in D-TBL group and objective item in the LBL group[(75.20 ± 11.82)％ vs.(68.61 ± 14.65)％,t=2.512,P=0.374].There was no statistically significant difference between the correct answer rate of group test in D-TBL group and objective item in the LBL group[(84.25 ± 13.08)％ vs.(68.61 ± 14.65)％,t=5.727,P=0.961].In Score table for members in every division,41(85.42％) students got straight A,7(14.58％) students got B and nobody got C.Feedback questionnaire showed 40(83.33％) students like D-TBL while 26(50.00％) students like LBL.Conclusion Most of students in D-TBL group like D-TBL.D-TBL and LBL cannot take the place of each other.In the future teaching,both methods should be used in different teaching periods according to their merits.

Objective To investigate the effects of hypoxia-inducible factor-1α (HIF-1α)expression on pathogenesis of retinopathy of prematurity (ROP) and to find new target for gene therapy.Methods After liposome-mediated small interference RNA (siRNA) transfection into rat retinal endothelial cells,the cells were cultured in medium with CoCl2-induced hypoxic condition.Expression of HIF-1α mRNA was determined by fluorenscence quantitative reverse transcription-polymerase chain reaction(RT-PCR),HIF-1α protein expression was detected by Western Blot after cocultured for 8 hours.Cell proliferation was measured with 3-(4,5)-dimethylthiazol (-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay after cocultured for 24 hours.Difference between groups was compared with independent samples t test.Results Rat retinal vascular endothelial cells were successfully transfected with siRNA.Fluorescence quantitative RT-PCR results showed that at 48 hours of transfection,the expression of HIF-1α mRNA in the interference group of siRNA1,siRNA2 and siRNA4 were 0.1620 ± 0.0147,0.2034 ± 0.0251 and 0.3049 ± 0.0165,which were 16.20％,20.34 ％ and 30.49％ of blank control group (1.0000±0.0344),and were lower than that of negative control group (0.8334±0.0242) (t=16.786,8.953 and 4.087,P＜0.05 respectively).Western Blot results showed that HIF-1α protein expression was significantly inhibited by siRNA1(0.4956 ± 0.0421 ) and siRNA2 (0.6544 ± 1.0032) comparing with blank control group (3.5105 ±0.4084) and negative control group (3.4019 ± 1.0677) (t =6.861,2.893,4.567 and 5.072,P＜0.05 respectively).As for cellular proliferation activity,(49.5±2.9) ％ and (67.4±1.2) ％ of cells growth inhibition were observed after transfection with siRNA1 and siRNA2,which were higher than those of negative control group [(15.7±1.5) ％ ] (t=2.786 and 6.904,P＜0.05).Conclusions The synthetic HIF-1α siRNA could effectively inhibit the expression of HIF-1α gene and reduce cell proliferation in rat retinal endothelial cells under hypoxic condition.RNA interference technology targeting HIF-1α might become a new strategy for gene therapy of ROP.

Objective To investigate the effect of rhubarb on hyperoxia-induced new bronchopulmo-nary dysplasia ( BPD) in rats. Methods Full-term Sprague-Dawley rats were exposed or not ( control group) in 600 ml/ L oxygen 4 days after birth and injected with normal saline (BPD group),rhubarb (BPD+ rhubarb group) or a combination of rhubarb with quercetin (BPD + rhubarb + quercetin group). Immuno-histochemical staining was used to detect the pathological changes of the rat lungs. HSP70 expression level was quantified by western blot. Results Compared with control group,BPD group showed decreased radial alveolar 14 and 21 days after the drug treatment,which was rescued by the coexistence of rhubarb. Quercetin, as an inhibitor of HSP70,counteracted the effect of rhubarb. The lung of the BPD + rhubarb + quercetin group showed a similar phenotypic change with that of the BPD group. In addition,the expressions of HSP70 in lung tissues of rhubarb group at 14 days and 21 days after the treatment were higher than those of other groups, there were significant differences between rhubarb group and other groups(F = 62. 46,P < 0. 01;F = 95. 90, P < 0. 01). Conclusion Rhubarb may attenuate the hyperoxia-induced new bronchopulmonary dysplasia in rats by activating HSP70 expression.

Objective:To investigate the impact of exogenous lactate on the replication of dengue virus type 2 (DENV-2) in Raw264.7 cells, mouse bone marrow-derived macrophages (BMDMs) and THP-1 cells and explore its association with cell activation.Methods:BMDMs from BALB/c mouse bone marrow were prepared and evaluated by flow cytometry to detect the proportion of F4/80 + CD11b + cells. Glucose transporter type 1 (GLUT1), hexokinase 2 (HK2), and monocarboxylate transporters 4 (MCT4) expression at mRNA level in BMDMs at different time points after DENV-2 infection were measured by qRT-PCR. The content of lactate in the culture supernatants was quantified via colorimetric assay. CCK-8 assay was used to evaluate the impacts of different concentrations of lactate on the viability of Raw264.7 cells, BMDMs, and THP-1 cells. qRT-PCR was used to detect the expression of DENV-2 E gene, TGF-β, CD86, retinoic acid-inducible gene Ⅰ (RIG-Ⅰ), IFN-β, interferon-stimulated gene 15 (ISG15), and ISG56 at mRNA level in cells infected with DENV-2 at different MOIs in the presence of different concentrations of lactate. Meanwhile, flow cytometry was used to analyze the expression of CD86 and CD206. Results:The percentage of BMDMs was (87.53±1.66)%. GLUT1 expression at mRNA level exhibited a decrease in BMDMs at 24 h after DENV-2 (MOI=3) infection following a transient increase at 12 h ( P<0.05), while HK2 expression at mRNA level was higher that than in blank control and inactivated DENV-2 infection groups at 12, 24, and 36 h ( P<0.01). Besides, there was an increase in both MCT4 mRNA level and the content of lactate in culture supernatants at 24 h after DENV-2 (MOI=1.5) infection ( P<0.05). The viability of the three types of cells remained above 80% when the concentration of lactate was 31.25 mmol/L. Lactate at the concentration of 35 mmol/L increased the expression of the DENV E gene at mRNA level in DENV-2-infected BMDMs at MOI=1 or MOI=2 ( P<0.05). Besides, it promoted the expression of DENV E gene at mRNA level in Raw264.7 and THP-1 cells ( P<0.001) as well as the expression of CD163, TGF-β, RIG-Ⅰ, IFN-β, ISG15 and ISG56 at mRNA level in BMDMs at MOI=1.5, but inhibited the expression of CD86 at mRNA level in BMDMs ( P<0.05). It also up-regulated CD206 protein expression ( P<0.01) and down-regulated CD86 protein expression ( P>0.05) in BMDMs. Conclusions:Exogenous lactate enhances DENV-2 replication in both human- and murine-derived macrophages and that might correlate with M2 macrophage polarization.

The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics. Drugging the multi-functional papain-like protease (PLpro) domain of the viral nsp3 holds promise. However, none of the known coronavirus PLpro inhibitors has been shown to be in vivo active. Herein, we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity, including against the Sarbecoviruses (SARS-CoV-1 and SARS-CoV-2), Merbecovirus (MERS-CoV), as well as the Alphacoronavirus (hCoV-229E and hCoV-OC43). Importantly, F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice. F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage, as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity. Despite the significant difference of substrate recognition, mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue, whereas an allosteric inhibitor of MERS-PLpro interacting with its 271E position. Our proof-of-concept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anti-coronavirus agents. The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.
Animals ; Coronavirus Papain-Like Proteases/antagonists & inhibitors* ; Cricetinae ; Humans ; Mice ; Pandemics ; SARS-CoV-2/enzymology* ; COVID-19 Drug Treatment