Background: Black bamboo root is traditionally used in the treatment of neurological injuries with definite efficacy. The study aimed to investigate the efficacy, and mechanism of action of black bamboo root extract quercetin on inflammation, apoptosis, and oxidation in rats with focal cerebral infarction. Methods: We used 64 adult male Sprague-Dawley rats to replicate a focal cerebral infarction model using the wire bolus method. The treatment group underwent pharmacological intervention and the control group was injected with the Toll-like receptor 4 (TLR4) blocker TAK242. A focal cerebral infarction model was created using middle cerebral artery occlusion in SpragueDawley rats. The immunofluorescence double-labelling method was employed to ascertain the expression of Glial Fibrillary Acidic Protein (GFAP), Ionized calcium-binding adaptor molecule 1 (Iba1) and TLR4 in glial cells. The expression of Neuron specific enolase (NSE) in neurons was determined through immunofluorescence. Immunohistochemistry was employed for the detection of cortical GFAP and Iba1 proteins. An immunoprotein blotting assay was employed to detect the presence of GFAP, Iba1, Nogo Receptor 3 (NgR3), Neurite outgrowth inhibitor-A (Nogo-A), Nuclear transcription factor NF-kappa B p65 (NF-kappa Bp65) and TLR4. The concentration of Interleukin17 (IL-17) was determined by enzyme-linked immunosorbent assay (ELISA). The activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) was measured by colorimetric assay, and the concentration of malondialdehyde (MDA) was determined by thiobarbituric acid assay (TBA). Results: The drug-treated group displayed a decrease in astrocyte GFAP and microglia iba1 with TLR4, neuronal NgR3 and Nogo-A expression; an increase in oxidase activity such as CAT, GSH-Px and SOD and a decrease in MDA expression. TLR4 receptor, NF-kB and IL-17 expression were decreased in control group after TLR4 inhibitor injection. Conclusion: Our findings suggest that black bamboo root extract has anti-inflammatory, and antioxidant effects in rats with cerebral ischaemia-reperfusion injury. The mechanism of action may involve the inhibition of TLR4 in astrocytes and microglia.

In recent years, inflammatory disorders have emerged as a significant concern for human health. Through ongoing research on anti-inflammatory agents, alpinetin has shown promising anti-inflammatory properties, including involvement in epigenetic modification pathways. As a crucial regulator of epigenetic modifications, Mecp2 may play a role in modulating the epigenetic effects of alpinetin, potentially impacting its anti-inflammatory properties. To test this hypothesis, two key components, p65 (a member of NF-KB family) and p300 (a type of co-activator), were screened by the expression profiling microarray, which exhibited a strong correlation with the intensity of LPS stimulation in mouse macrophages. Meanwhile, alpinetin demonstrates the anti-inflammatory properties through its ability to disrupt the synthesis of p65 and its interaction with promoters of inflammatory genes, yet it did not exhibit similar effects on p300. Additionally, Mecp2 can inhibit the binding of p300 by attaching to the methylated inflammatory gene promoter induced by alpinetin, leading to obstacles in promoter acetylation and subsequently impacting the binding of p65, ultimately enhancing the anti-inflammatory capabilities of alpinetin. Similarly, in a sepsis mouse model, it was observed that homozygotes overexpressing Mecp2 showed a greater reduction in organ damage and improved survival rates compared to heterozygotes when administered by alpinetin. However, blocking the expression of DNA methyltransferase 3A (DNMT3A) resulted in the loss of Mecp2's anti-inflammatory assistance. In conclusion, Mecp2 may augment the anti-inflammatory effects of alpinetin through epigenetic 'crosstalk', highlighting the potential efficacy of a combined therapeutic strategy involving Mecp2 and alpinetin for anti-inflammatory intervention.

Objective To explore the application effect of BOPPPS model + network course platform in internal science teaching. Methods A total of 116 clinical medical undergraduates in grade 2023 of College of Adult Continuing Education of Hunan University of Medicine were randomly divided into experimental class and control class with 58 students in each class by numerical table method. From April to May 2024, BOPPPS+ network course platform teaching will be implemented in the experimental class, while traditional teaching will be carried out in the control class. At the end of the experiment, the teaching design satisfaction survey, independent learning ability test and internal medicine learning level test were carried out. SPSS 20.0 software was used for statistical analysis, Chi-square test was used for counting data comparison between two classes, and T-test was used for measurement data comparison. Results The satisfaction of teaching design in the experimental class was 50 (86.2%), which was significantly higher than that in the control class 43 (74.1%), the difference was statistically significant. The total score of independent learning in the experimental class (254.3±6.7) was significantly higher than that of the control class (236.0±7.4), and the differences were statistically significant. The scores of internal medicine test, theory test and total score of experimental class were (35.8±2.2)(46.7+5.1)(81.6±2.6) points respectively, which were significantly higher than that of control class (34.8±2.2)(43.4±4.3)(78.6+4.7) points, with statistical differences. Conclusion BOPPPS model+ network course platform teaching can greatly improve students' satisfaction, autonomous learning ability and internal medicine learning level.

Abstract Traditional periodontal therapy primarily controls the progression of chronic periodontitis but falls short of fully regenerating periodontal tissues. Selecting suitable seed cells, growth factors, and ideal scaffold materials is crucial for maximizing periodontal tissue regeneration, a key factor in the success of periodontal tissue repair. This study involved constructing FGF2 chitosan nanospheres (single-shell packaging) and BMP9 chitosan nanospheres hydrogel (double-shell packaging) via an ion crosslinking method. By examining the surface morphology, particle diameter, drug loading rate, and encapsulation efficiency of these nanospheres, it was confirmed that they satisfy the criteria for nanomaterial sustained release carriers. Additionally, they effectively manage the initial "burst release" by enabling the controlled and gradual release of FGF2 and BMP9 over a specific period. The FGF2-PLGA/PLGA-nHA-BMP9 biphasic scaffold, developed using 3D printing technology in conjunction with the aforementioned nanospheres, was assessed for biomechanical properties and absorbance metrics such as compressive strength, tensile strength, and elastic modulus. The scaffold's cell adhesion and proliferation were evaluated using the MTT assay, while genes associated with fibroblast differentiation (SCX, Col-I) and osteogenic differentiation (Runx-2, ALP) were analyzed through qRT-PCR. The findings confirm that the 3D-printed FGF2-PLGA/PLGA-nHA-BMP9 biphasic scaffold possesses excellent biomechanical properties and biocompatibility. It effectively orchestrates the sequential release of FGF2 and BMP9 in distinct scaffold phases, synergistically enhancing periodontal tissue regeneration and maximizing the biological activity of FGF2 and BMP9.

Tetrahydroxy stilbene glucoside (TSG) from Polygonum multiflorum exerts neuroprotective effects after ischemic stroke. We explored whether TSG improved ischemic stroke injury via PTEN-induced kinase 1 (PINK1)/Parkin-mediated mitophagy. Oxygen glucose deprivation/reoxygenation (OGD/R) in vitro model and middle cerebral artery occlusion (MCAO) rat model were established. Cerebral injury was assessed by neurological score, hematoxylin and eosin staining, 2,3,5-triphenyltetrazolium chloride staining, and brain water content. Apoptosis, cell viability, and mitochondrial membrane potential were assessed by flow cytometry, cell counting kit-8, and JC-1 staining, respectively. Colocalization of LC3-labeled autophagosomes with lysosome-associated membrane glycoprotein 2-labeled lysosomes or translocase of outer mitochondrial membrane 20-labeled mitochondria was observed with fluorescence microscopy. The ubiquitination level was determined using ubiquitination assay. The interaction between molecules was validated by coimmunoprecipitation and glutathione S-transferase pull-down. We found that TSG promoted mitophagy and improved cerebral ischemia/reperfusion damage in MCAO rats. In OGD/R-subjected neurons, TSG promoted mitophagy, repressed neuronal apoptosis, upregulated Y-box binding protein-1 (YBX1), and activated PINK1/Parkin signaling. TSG upregulated ubiquitin-specific peptidase 10 (USP10) to elevate YBX1 protein. Furthermore, USP10 inhibited ubiquitination-dependent YBX1 degradation. USP10 overexpression activated PINK1/Parkin signaling and promoted mitophagy, which were reversed by YBX1 knockdown. Moreover, TSG upregulated USP10 to promote mitophagy and inhibited neuronal apoptosis. Collectively, TSG facilitated PINK1/Parkin pathway-mediated mitophagy by upregulating USP10/YBX1 axis to ameliorate ischemic stroke.

Long noncoding RNAs (LncRNAs) are essential to regulate the pathogenesis of coronary artery disease (CAD). This study was conducted to analyze the functionality of long noncoding RNA cancer susceptibility candidate 11 (lncRNA CASC11) in oxidized low-density lipoprotein (ox-LDL)-induced injury of cardiac microvascular endothelial cells (CMECs). CMECs were treated with ox-LDL to induce the CAD cell model. The cellular expression levels of CASC11 and histone deacetylase 4 (HDAC4) were determined by real-time quantitative polymerase chain reaction or Western blot assay. Cell absorbance, apoptosis, angiogenesis, and inflammation were evaluated by cell counting kit-8, flow cytometry, tube formation, and enzyme-linked immunosorbent assays. The subcellular localization of CASC11 was examined by the nuclear/cytoplasmic fractionation assay. The binding of human antigen R (HuR) to CASC11 and HDAC4 was analyzed by RNA immunoprecipitation. HDAC4 stability was determined after actinomycin D treatment. CASC11 was found to be decreased in the CAD cell model. CASC11 upregulation increased cell viability and angiogenesis and reduced apoptosis and inflammation. CASC11 bound to HuR and improved HDAC4 expression. HDAC4 downregulation counteracted the protective role of CASC11 overexpression in CMECs. In summary, CASC11 alleviated ox-LDL-induced injury of CMECs by binding to HuR and stabilizing HDAC4.

In this study, we investigated the effect of sweroside (SOS) on hepatic steatosis in mice and elucidated its molecular mechanisms. We conducted in vivo experiments using a C57BL/6 mice model of nonalcohol fatty liver disease (NAFLD) to explore the effect of SOS on hepatic steatosis in NAFLD mice. In in vitro experiments, primary mouse hepatocytes were treated with palmitic acid and SOS, and the protective effects of SOS on inflammation, lipogenesis, and fat deposition were analyzed. Autophagy-related protein levels and their related signaling pathways were evaluated in both in vivo and in vitro experiments. The results demonstrated that SOS decreased the high-fat-induced intrahepatic lipid content both in vivo and in vitro. The autophagy level in the liver was decreased in NAFLD mice but was reactivated following SOS intervention. SOS intervention was found to partially activate autophagy via the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Consequently, when the AMPK/mTOR pathway was suppressed or autophagy was inhibited, the beneficial effects of SOS intervention on hepatic steatosis were diminished. These results indicate that SOS intervention attenuates hepatic steatosis by promoting autophagy in the liver of NAFLD mice, in part by activating the AMPK/mTOR signaling pathway.

In this study, we investigated the effects of sweroside on podocyte injury in diabetic nephropathy (DN) mice and elucidated its molecular mechanisms. We conducted in vivo experiments using a C57BL/6 mice model of DN to explore the effects of sweroside on proteinuria and podocyte injury in DN mice. In in vitro experiments, conditionally immortalized mouse podocytes were treated with high glucose and sweroside, and the protective effects of sweroside on podocyte injury were analyzed. In vitro, Akt/BAD pathways were detected using gene siRNA silencing assays and found to be involved in the protective roles of sweroside in high glucose-mediated podocyte injury. In vivo, sweroside significantly decreased albuminuria in DN mice (p < 0.01). periodic acid-Schiff staining showed that sweroside alleviated the glomerular volume and mesangium expansion in DN mice. Consistently, western blot and reverse transcription-polymerase chain reaction analyses showed that the profibrotic molecule expression in the glomeruli declined in sweroside-treated DN mice. Immunofluorescent results showed that sweroside preserved nephrin and podocin expression, and transmission electron microscopy showed that sweroside attenuated podocyte injury. In DN mice, sweroside decreased podocyte apoptosis, and increased nephrin, podocin expression and decreased desmin and HIF1α expression. These results confirmed that sweroside ameliorated albuminuria, glomerulomegaly, and glomerulosclerosis in these mice. Experiments in vitro revealed that sweroside improved HG-induced podocyte injury and apoptosis. Sweroside stimulated activation of the Akt/BAD pathway and upregulated Bcl-2-associated death promoter (BAD) and p-Akt. Overall, sweroside protected podocytes from injury and prevented the progression of DN, providing a novel strategy for the treatment of DN.

This study aimed to analyze the effect of emodin on alveolar bone resorption in rats with periodontitis through the interleukin-23 (IL-23)/T helper 17 (Th17) cells inflammatory axis. Twenty-four male Sprague Dawley rats were randomly divided into the following three groups of eight rats each: the control group (group N), periodontitis group (group P), and periodontitis + emodin intervention group (group E). After 2 weeks of continuous emodin intervention, alveolar bone loss (ABL) was evaluated by morphological analysis. Tartrate-resistant acid phosphatase staining was performed to determine the number of osteoclasts (OCs) in periodontal tissue. The ABL and trabecular separation were greater and the number of OCs and the G1/G0 and G2/M phase ratios were higher in group P than those in groups N and E (P < 0.05). In contrast, the trabecular number and thickness were decreased and the S and G2+S phase ratios were lower in group P than those in groups N and E (P < 0.05). However, these indices did not differ significantly between groups E and N. The IL-23, IL-17, RANKL, RANK, RANKL/OPG, IL-2, IL-6, IL-1 beta, and TNF-alpha levels across the three groups were in the order of group P > group E > group N and the OPG levels were in the order of group P < group E < group N (P < 0.05). Emodin may decrease the expression of RANKL and RANK proteins and levels of inflammatory factors; increase OPG, PPAR-gamma, and NF-kappa B protein expression levels; and decrease the number of OCs through the IL-23/Th17 inflammatory axis, thus inhibiting ABL and bone resorption in rats with periodontitis and promoting the proliferation of periodontal ligament cells and alveolar bone repair.

Objective To investigate the underlying molecular mechanism of methyl-CpG-binding protein 2 (MeCP2) inhibiting interleukin 6 (IL-6) transcriptional activity by observing the sequence of methylated IL-6 promoter, overexpression of MeCP2, and transcription factor P300 in HEK293 cells. Methods The binding site of P300 in the IL-6 promoter region was confirmed by electrophoretic mobility shift assay (EMSA); the IL-6 promoter sequence was ligated into luciferase reporter plasmid and transfected into HEK293 cells. The methylation of the promoter was mediated by clustered regularly interspaced short palindromic repeats-deactivated Cas9 (CRISPR-dCas9)-mediated DNA methyltransferase 3A (DNMT3A) transfection, and then MeCP2 and P300 overexpression plasmids were transfected. The bisulfate sequencing PCR(BSP)was used to analyze the cytosine methylation in the IL-6 promoter region of each group. The contents of intracellular MeCP2 and P300 were detected by the Western blot. A chemiluminescence detector was used to determine the luciferase activity of HEK293 cells. The binding level of P300 and MeCP2 in the IL-6 promoter region was analyzed by chromatin immunoprecipitation followed by sequencing(ChIP-seq). Results EMSA confirmed the presence of P300 binding sites in the IL-6 promoter of mice. CRISPR-dCas9-DNMT3A transfection into HEK293 cells successfully methylated the IL-6 promoter. MeCP2 and P300 overexpression plasmid steadfastly synthesized the target protein and was not affected by other transfection. Compared with the unmodified promoter, methylation could reduce the transcriptional activity of the promoter. When P300 was overexpressed, MeCP2 could further inhibit the transcriptional activity of the promoter, when compared with methylation alone. Also, overexpression of P300 could not promote the transcriptional activity of IL-6 promoter after the methylation modified promoter combined with MeCP2, while the overexpression of P300 enhanced the transcriptional activity when the promoter was not methylated or MeCP2 was not overexpressed. ChIP-seq analysis revealed that the methylated IL-6 promoter showed no difference in binding to P300; however, when combined with MeCP2, the binding capacity would be repressed. Conclusion The combination of MeCP2 with methylated IL-6 promoter can inhibit the binding of the transcription factor to the promoter, thereby impeding the transcriptional activity of the promoter.

Objective To explore the cultivation and application effect of critical thinking ability in the class teaching of clinical-thinking training for diagnostics. Method To select as teaching objects 2 groups of junior students who major in clinical medicine in a 5-year program at Hunan University of Medicine — Group 1 with 182 students and Group 2 with 178 students. Group 1 functions as a control group and Group 2 an experimental group. The control group uses both the traditional teaching and classroom discussion methods for teaching, while the experimental group uses the method of “real cases+ critical thinking” for teaching. Before the class, the students in the experimental group are provided with two cases for their preview work; during the class, they are introduced into case scenes through videos; after the class, the teaching effects of the 2 groups are evaluated through the chapter tests, CTDI-CV scores, and questionnaires. The survey data are statistically analyzed with SPSS20.0 software, and the t test is used for comparison between the two groups. Results The chapter test and the critical thinking ability test for the students in the experimental group score in total respectively (87.17±6.56) and (301.05±29.58), both of which are higher than those of the control group (the two kinds of scores for the control group are (79.27±7.36) and (254.23±24.74) respectively), with the differences statistically significant (P<0.05). The questionnaire-response rate of the experimental group students stands at 100%; most students believe that “real cases + critical thinking” in the class teaching could improve students' clinical-thinking ability in diagnosing diseases, cultivate students’ good character of selfless dedication, and familiarize students with the clinical characteristics of related diseases. Conclusion In the teaching process of clinical-thinking training for diagnostics, applying both the real cases and critical thinking is conducive to improving the teaching effect, and therefore it is worth promoting in clinical teaching and practice.

Background:Recently, inflammation has become a major threat to human health. Studies have confirmed that some Chinese traditional medicine ingredients may effectively interfere with the expression of inflammatory mediators through epigenetic modification, showing a great potential of the application.Objective:To investigate the role of the PPAR/DNMT3A pathway in the reversal of galangin-mediated inflammatory lung injury, promote the development of new anti-inflammatory drugs, reduce the side effects of chemical synthetic drugs on the body, and prove the effectiveness and safety of galangin in inhibiting inflammatory response and injury.Methods:120 rats were randomly divided into 6 groups: (Group 1) LPS group; (Group 2) LPS + galangin group; (Group 3) LPS + galangin +  GW9662 group; (Group 4) LPS + galangin + DNMT3A siRNA group; (Group 5) LPS + galangin + siRNA negative group; (Group 6) control group. The model of inflammatory lung injury was established by intrathecal instillation of LPS in the first five groups and NS in the control group. SD survival rate was recorded every 24 hours after modeling, lasting for 168 hours. The lung tissues were taken 168 hours after the establishment of the model. The pathological morphology of lung tissue was observed after the staining under the light microscope, and the lung dry/wet weight ratio was calculated after drying. After NS was perfused into lung tissue, the lavage fluid was collected and the levels of IL-6 and TNF-a were measured by ELISA. The contents of PPAR, DNMT3A, phosphorylated p65, and ERK in monocytes were detected by the WB method, and the binding contents of p65 and AP-1 in the promoter regions of IL-6 and TNF-a genes were detected by the Chip-qPCR method.Results:Intraperitoneal injection of galangin could inhibit the synthesis of alveolar inflammatory factors (TFs) in the SD model of lung injury induced by LPS, reduce the degree of pathological injury of lung tissue, and improve the survival rate of the SD model. GW9662 can completely reverse the protective effect, while DNMT3A interference can only partially block its protective effect. In addition, galangin could significantly inhibit the LPS-induced expression of p65 and AP-1 in alveolar monocytes and their binding content in the promoter region of inflammatory genes by activating PPAR/DNMT3A pathway. GW9662 could completely reverse the inhibitory effect of galangin. DNMT3A interference could restore the binding content of transcription factors at the promoter of the inflammatory gene but had no significant effect on its synthesis.Conclusion:Galangin can interfere with the binding of transcription factors to inflammatory gene promoters through the methylation modification induced by PPAR/DNMT3A pathway, so as to inhibit the synthesis of inflammatory molecules and reverse inflammatory lung injury.

INTRODUCTION:The study aimed to explore the effects of treatment with black bamboo rhizome extracts on learning and memory and determine the underlying mechanisms in rats with cerebral ischaemia-reperfusion injury.METHODS:Sprague-Dawley rats were randomly divided into the following four groups: control, middle cerebral artery occlusion (MCAO), low-dose drug, and high-dose drug groups. Rats underwent MCAO using a suture method before drug treatment. Then, neurological impairment was assessed using the Longa scoring method, and triphenyl tetrazolium chloride staining was used to analyse the cerebral infarction area. The Elliott formula was used to calculate water content in the brain tissue. A Morris water maze (MWM) was used to assess changes in learning and memory abilities, and Western blotting was used to detect cyclic adenosine phosphate response element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) expression in the hippocampus of MCAO rats.RESULTS:After treatment with black bamboo rhizome extracts, the neurological dysfunction score was lower in the drug groups than in the MCAO group, and a significant difference was observed between the high-dose drug and MCAO groups (P<0.05). Additionally, the cerebral infarction area was significantly smaller in the drug groups than in the MCAO group (P<0.01), and the effect was more obvious in the high-dose drug group than in the low-dose drug group. There was also a significant difference in water content between the high-dose drug and MCAO groups, and cerebral oedema was significantly reduced in the high-dose drug group (P<0.05). In the MWM, the incubation period was significantly reduced, the number of platform crossings was significantly increased, and the search time was prolonged in the drug groups compared with those in the MCAO group (P<0.05). Moreover, the expression of BDNF and CREB was significantly increased in the drug groups compared to that in the MCAO group, and the increase was more obvious in the high-dose group than in the low-dose group (P<0.05).DISCUSSION:Black bamboo rhizome extracts significantly improved cognitive dysfunction, reduced cerebral oedema, decreased the cerebral infarction area, and improved the neurological function score and learning and memory abilities in rats with cerebral ischaemia-reperfusion injury.

Objective:To evaluate the effect of teaching method based on online resource-sharing mechanism in diagnostic teaching.Methods:From September, 2019 to July, 2020, totally 720 students of 5-year curriculum of clinical medicine enrolled in 2017 in Hunan University of Medicine were selected and divided into experimental group and control group with 360 students in each. In the teaching of diagnostics, off-line teaching combined with online resource-sharing approach was adopted in the experimental group, while off-line traditional teaching approach was adopted in the control group. At the end of the teaching, theoretical examination method was used to compare the results of the two groups , and the questionnaire survey was used to investigate the experimental group students' evaluation on the effect of online resource-sharing method.Results:Both midterm and final test scores of the experimental group [(83.25±5.72) and (85.14±7.65)] were significantly higher than those of the control group [(78.02±5.39) and (77.36±7.45)](all P<0.01). The results of the questionnaire survey showed that 85.0%(306/360) of the students in the experimental group believed that the network resource-sharing method in teaching of diagnostics greatly improved their autonomic learning capacity. Conclusions:The teaching approaches of online resource sharing diagnostic course improves the learning outcomes and autonomic learning attitude of students.

INTRODUCTION:Melatonin (Mel) and its receptors are promising for glycaemic control in patients with type 2 diabetes mellitus (T2DM) and its complications, but there is significant heterogeneity among studies. This study aims to investigate the effects of Mel receptor agonist Neu-P11 on glucose metabolism, immunity, and islet function in T2DM rats. MATERIAL AND METHODS:In this study, SD rats were treated with a high-fat diet and streptozotocin (STZ) to establish a T2DM model. The glucose oxidase method was used to measure blood glucose levels. Glucose and insulin tolerance tests were used to assess glucose metabolism. Haematoxylin-eosin staining was used to observe pancreatic tissue injury. The apoptosis of isletβ cells was analysed by TUNEL and insulin staining. Reactive oxygen species (ROS) levels and immune cell expression were analysed by flow cytometry. IF was used to analyse the activation of microglia. The immunoglobulins: IgA, IgG, IgM, tumour necrosis factorα (TNF-α), interleukins IL-10 and IL-1β, interferonγ (IFN-γ), C-peptide, and insulin levels were determined by ELISA. The expression of CD11b, CD86, cleaved caspase 3, p21, and P16 proteins were analysed by western blot. RESULTS:The results showed that the blood glucose level increased, insulin resistance occurred, spleen coefficient and ROS levels increased, humoral immunity in peripheral blood decreased, and inflammation increased in the model group compared to the control group. After Mel and Neu-P11 treatment, the blood glucose level decreased significantly, insulin sensitivity improved, spleen coefficient and ROS levels decreased, humoral immunity in peripheral blood was enhanced, and inflammation improved in T2DM rats. Brain functional analysis of T2DM rats showed that microglia cells were activated, TNF-α and IL-β levels were increased, and IL-10 levels were decreased. Mel and Neu-P11 treatment reversed these indexes. Functional analysis of islets in T2DM rats showed that islet structure inflammation was impaired, isletβ cells were apoptotic, p21 and p16 protein expressions were increased, and blood C-peptide and insulin were decreased. Mel and Neu-P11 treatment restored the function of pancreatic b cells and improved the damage of pancreatic tissue. CONCLUSION:Melatonin and its receptor Neu-P11 can reduce the blood glucose level, enhance humoral and cellular immunity, inhibit microglia activation and inflammation, and repair isletβ cell function, and this improve the characterization of T2DM-related diseases.

Background Ischemic stroke, also known as cerebrovascular accident or cerebral stroke, occupies the first place in the world's top 10 causes of death, with high incidence, mortality and disability rates. Objectives To investigate the effect of stilbene glycoside upregulated SIRT3/AMPK expression on neuronal mitochondrial autophagy and neuronal apoptosis in ischemic stroke. Material and methods The PC12 cells were cultured without serum to construct an ischemic neuron model. The cells were divided into 6 groups: normal group (untreated cells), model group (ischemic treated cells), TSG group (stilbene glycoside treatment), NC group (SIRT3 and AMPK negative control treatment), si-SIRT3 group (SIRT3 silencing treatment), TSG+si-SIRT3 group (joint treatment), and TSG+si-SIRT3+oe-AMPK group (joint treatment). Cell survival and the expression of related molecules were detected. Results Compared with normal group, the model group had significantly decreased cell survival rate, mitochondrial membrane potential, as well as the expression of Bcl-2, LC3II/I, P62, PINK1, Parkin, SIRT3, AMPK, and p-AMPK, while showing significantly increased proportion of apoptosis and the expression of caspase 3 and Bax. Compared with the model group, TSG treatment promoted cell survival rate and mitochondrial autophagy, and inhibited apoptosis, while SIRT3 silencing treatment reduced cell survival rate and mitochondrial autophagy, and increased apoptosis. The SIRT3 silencing could block the inhibitory effect of TSG on the apoptosis of ischemic PC12 cells and promote mitochondrial autophagy, and AMPK overexpression could save the apoptosis of ischemic PC12 cells caused by SIRT3 silencing, and promote mitochondrial autophagy. Conclusions By promoting the expression of SIRT3/AMPK, TSG promotes mitochondrial autophagy in ischemic neurons and inhibits their apoptosis.

随着信息化教学改革的不断发展,医学高校线上线下教学模式应用越来越广泛,通过对医学高校线上、线下、线上线下混合式教学模式的利弊探讨,发现单一的线上教学或线下教学由于存在一些弊端,已难以满足教学和学生的需求.而线上线下相结合的混合式教学模式能让"教师动起来,学生活起来",通过改革教学方法和手段、完善教师培训机制、加强课程建设、构建线上线下课程成绩考核评价体系开展线上线下混合式教学,相较于传统教学模式,能更有效地提高教学质量.

目的 通过运用行为阶段模型(TTM)对医学生临床技能学课程学习动机进行评估,探索虚拟仿真教学方法的积极作用.方法 选取该校处于本科第3学年2学期临床医学专业学生240名,在均衡原则下随机分配入虚拟仿真教学组及传统教学组,每组120名.进入分组前和学期末,学生完善T T M评估量表;同时搜集学期末临床技能诊断部分的实践考核成绩进行统计学分析.结果 学期末,虚拟仿真教学组学习动机处于第四、五阶段学习阶段学生所占比例优于传统教学组,差异有统计学意义(P<0.05).虚拟仿真教学组平均成绩[(80.13±3.32)分]高于传统教学组[(74.26±4.25)分],差异有统计学意义(t=6.42,P<0.05).结论 TTM直观显示了虚拟仿真教学方法在提升医学生学习动机方面的优势,值得在教学评价实践中推广.

目的 通过检测山姜素对鼠巨噬细胞(RAW246.7) IL-6启动子部位CpG岛二核苷酸甲基化修饰状态及IL-6合成的影响,揭示诱导甲基化修饰是山姜素调控炎症分子表达的重要机制.方法 RAW246.7按照不同干预分为对照组、山姜素组(终质量浓度分别为100、200、500 μg/ml以及1 mg/ml)、山姜素+GW9662组以及山姜素+DNMT3A(DNA甲基转移酶3A)SiRNA组,孵育96 h后经亚硫酸氢钠处理.亚硫酸氢盐测序PCR分析IL-6启动子序列碱基信息,甲基化PCR分析以上序列中CpG二核苷酸甲基化修饰状态,Western blot法检测RAW246.7核内PPAR以及DNMT3A含量,ELISA法检测IL-6表达水平.结果 经Cpgplot数据库确认鼠巨噬细胞IL-6转录起始点上游300～1 300 bp部位存在典型CpG岛,山姜素可呈剂量依赖性促进该岛内CpG二核苷酸甲基化修饰.山姜素可促进核内DNMT3A合成,GW9662可阻断促进作用,而干扰DNMT3A不影响PPAR表达;另外,GW9662以及DNMT3A干扰均可完全逆转山姜素对甲基化修饰的促进;但GW9662更明显恢复了IL-6的合成.结论 山姜素可通过PPAR/DNMT3A通路促进RAW246.7IL-6启动子CpG岛内CpG二核苷酸甲基化修饰,从而抑制IL-6表达.

通过分析甲基CpG结合蛋白2(methyl CpG binding protein 2,MeCP2)对山姜素作用下鼠巨噬细胞(RAW246.7)合成IL-6和TNF-α的影响及作用机制,揭示山姜素与MeCP2联合应用对于炎症性疾病的干预潜力.用重组质粒pEGFP-C1-MeCP2以及空白质粒pGL-basic分别转染RAW246.7,后将细胞随机分为对照组、山姜素组(终浓度1 mg/mL)以及山姜素+GW9662(0.1 mmol/L)组.孵育72 h后,ELISA检测培养液中IL-6和TNF-α表达水平,Western blotting分别检测细胞核内过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptor,PPAR)、MeCP2以及胞质中NLRP3、pro-Caspase-1和各聚集状态凋亡相关微粒蛋白(apoptosis-associated speck-like protein containing CARD,ASC)含量,并采用免疫荧光染色法检测ASC斑点形成情况.结果 显示,与空白质粒转染比较,重组质粒转染RAW246.7在山姜素作用下合成IL-6和TNF-α的水平更低;而在对照组以及GW9662组,转染重组质粒对炎性因子合成无明显影响.山姜素对MeCP2表达无促进作用,且重组质粒转染对PPAR表达亦无影响.另外,山姜素可明显抑制RAW246.7胞质NLRP3合成,但对pro-Caspase-1含量以及ASC聚集状态无显著影响;而过表达MeCP2可遏制pro-Caspase-1合成并促进ASC解聚.由此,MeCP2可通过抑制炎症小体通路以促进山姜素对鼠巨噬细胞炎性因子合成的抑制,提示山姜素与MeCP2联合有望成为炎症性疾病防治的新策略.