Osteoarthritis (OA) is a leading cause of pain and disability worldwide. Despite extensive research, no curative treatment is currently available. Most therapies focus on symptom relief or disease modification. Traditional Chinese medicines, particularly Astragalus mongholicus (Huangqi) and Saposhnikovia divaricata (Fangfeng), have demonstrated therapeutic potential for OA, but their molecular mechanisms remain unclear. The synovium plays a key role in OA pathogenesis and disease progression, yet the effects of these herbal extracts on synoviocytes are poorly understood. To explore potential targets, we analyzed all publicly available microarray datasets comparing gene expression in OA and healthy synovium/synoviocytes. Differentially expressed genes (DEGs) were identified and compared with known targets of A. mongholicus and S. divaricata. The intersecting genes were subjected to Gene Ontology, KEGG pathway, and protein‒protein interaction analyses to identify hub genes. Key transcription factors were predicted using the KNOCK.TF database. Expression of hub genes and transcription factors was validated in human synoviocytes treated with A. mongholicus and S. divaricata extracts. We identified ten hub genes, eight of which may serve as OA biomarkers. Three hub transcription factors—AR, CITED2, and SF1—were upregulated by the extracts, while MMP2 and MMP9, two OA-associated genes, were downregulated. Our findings suggest that A. mongholicus and S. divaricata may exert therapeutic effects on OA by modulating synoviocyte gene expression, notably through upregulation of AR and CITED2 and downregulation of MMP2 and MMP9. These herbs may provide molecular targets for novel OA treatments.

Objective Osteosarcoma is the most common primary bone cancer that affects mostly children and young adults. Despite the advances in osteosarcoma treatment, the long-term survival rate of metastatic patients has not significantly improved in the past few decades, thus demonstrating the need for novel therapeutic targets or methods to improve metastatic osteosarcoma treatment. In this study we aimed to elucidate the role of miR-659-3p and SRPK1 in osteosarcoma. Methods We evaluated miR-659-3p and SRPK1 function in osteosarcoma cell proliferation, migration, and cell cycle progression in vitro by using gain- and loss-of-function strategies. The effect of miR-659-3p in tumor progression and metastasis was determined by in vivo mouse model. Results We revealed that expression of miR-659-3p was significantly downregulated in osteosarcoma compared with normal bone cells and was inversely correlated with serine-arginine protein kinase 1 ( SRPK 1) expression. We proved that miR-659-3p targets 3’ UTR of SRPK1 and negatively regulates SRPK 1 expression in osteosarcoma cells via luciferase assay. In vitro studies revealed that gain of miR-659-3p function inhibited osteosarcoma cells growth, migration, and invasion by down-regulating SRPK 1 expression. Inversely, inhibiting miR-659-3p in osteosarcoma cells promoted cell growth, migration, and invasion. Cell cycle profile analysis revealed that miR-659-3p inhibited osteosarcoma cells’ G1/G0 phase exit by down-regulating SRPK 1 expression. By using an in vivo mouse model, we demonstrated that miR-659-3p inhibits osteosarcoma tumor progression and lung metastasis by inhibiting SRPK 1 expression and potentially downstream cell proliferation, and epithelial-to-mesenchymal transition genes. Conclusions This study demonstrated that miR-659-3p is a potential therapeutic method and SRPK1 is a potential therapeutic target for osteosarcoma treatment.

The expression of genes altered in epilepsy remains incomplete, particularly in the hippocampus, which exhibits exquisite vulnerability to epilepsy. Q808 is an innovation chemical compound that has potent anti-convulsant effect. Exploring its mechanism can not only explore the pathogenesis of epilepsy but also provide a theoretical basis for its clinical application. The present study aimed to use RNA sequencing (RNA-seq) to reveal the gene transcriptomic profile of chronic pentylenetetrazole (PTZ)-kindled seizure rats and the difference of the PTZ model rat before and after treatment with Q808. Quantitative real-time PCR (qRT-PCR) was performed to validate the RNA-seq results. The protein level was estimated with Western blot. Hippocampal transcriptomic analysis showed that 289 differentially expressed genes (DEGs) were confirmed in the PTZ-kindled seizure group compared with the vehicle control. Gene cluster analysis identified most of the DEGs linked to neuronal apoptosis, neurogenesis, neuronal projections, and neurotransmitter regulation. After analysis across the three groups, 23 hub genes and 21 pathways were identified, and qRT-PCR analysis confirmed that most of the mRNA levels of hub genes were consistent with the RNA-seq results. Q808 treatment increased the level of ACE, a GABA-related protein. Our analysis showed the comprehensive compendium of genes and pathways differentially expressed for PTZ-kindled seizure rats and upon Q808 treatment in PTZ-kindled seizure, which may provide a theoretical basis to explore the mechanism and unique efficacy of Q808 and the pathophysiology of epilepsy in the future.

IntroductionAKR1C3, as a crucial androgenic enzyme, implicates the androgen biosynthesis and promoting prostate cancer cell growth in vitro. This study provides a new gene therapy strategy for targeting AKR1C3 to treat castration-resistant prostate cancer.MethodssiAKR1C3@PPA is assembled from PEG3500, PAMAM, Aptamer-PSMA, and siRNA for AKR1C3. We analyzed the relationship between AKR1C3 expression and the survival rate of prostate cancer patients based on the GEPIA online database to perform disease-free survival, and found that AKR1C3 may be an important factor leading to poor prognosis in prostate cancer. Considering AKR1C3 as a therapeutic target for castration-resistant prostate cancer, we constructed a complex nucleic acid nanoparticle, siAKR1C3@PPA to investigate the inhibitory effect on castration-resistant prostate cancer.ResultsAptamer-PSMA acts as a target to guide siAKR1C3@PPA into PSMA-positive prostate cancer cells and specifically down regulate AKR1C3. Cyclin D1 was decreased as a result of siAKR1C3@PPA treatment. Changes in Cyclin D1 were consistent with decreased expression of AKR1C3 in LNCaP-AKR1C3 cells and 22RV1 cells. Furthermore, in the LNCaP-AKR1C3 group, 1070 proteins were upregulated and 1015 proteins were downregulated compared to the LNCaP group according to quantitative 4D label-free proteomics. We found 42 proteins involved in cell cycle regulation. In a validated experiment, we demonstrated that PCNP and CINP were up-regulated, and TERF2 and TP53 were down-regulated by western blotting.ConclusionWe concluded that siAKR1C3@PPA may arrest the cell cycle and affect cell proliferation.

Osteosarcoma (OS) is the most common type of primary bone tumor in children and adults. Dangshen (Codonopsis pilosula) is a traditional Chinese medicine commonly used in the treatment of OS worldwide. However, the molecular mechanisms of Dangshen in OS remain unclear. Hence, in this study, we aimed to systematically explore the underlying mechanisms of Dangshen in the treatment of OS. Our study adopted a network pharmacology approach, focusing on the identification of active ingredients, drug target prediction, gene collection, gene ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and other network tools. The network analysis identified 15 active compounds in Dangshen that were linked to 48 possible therapeutic targets related to OS. The results of the gene enrichment analysis show that Dangshen produces a therapeutic effect in OS likely by regulating multiple pathways associated with DNA damage, cell proliferation, apoptosis, invasion, and migration. Based on the network pharmacology approach, we successfully predicted the active compounds and their respective targets. In addition, we illustrated the molecular mechanisms that mediate the therapeutic effect of Dangshen in OS. These findings may aid in the development of novel targeted therapies for OS in the future.

Introduction: Osteosarcoma is the most common bone tumor and is characterized by the presence of malignant mesenchymal cells produced in the bone stroma. MiRNAs are known to function as post-transcriptional negative regulators of gene expression. Emerging evidence showed that miR-1225-5P functions as a tumor suppressor in several types of cancers. The detailed mechanisms of which miR-1225-5P suppresses tumor growth are not fully understood. The objective of the present study was to test the hypothesis that miR-1225-5P inhibits osteosarcoma cell growth in vitro and tumor growth in vivo by targeting YWHAZ expression. Methods: Real-time PCR and Western blot were carried out to test the expression of miR-1225-5P and YWHAZ in osteosarcoma cell lines. Luciferase assay was used to demonstrate whether miR-1225-5P targets YWHAZ 3' UTR. To assess the function of miR-1225-5P in human osteosarcoma cell lines, gain-of-function and loss-of-function of miR-1225-5P were performed by transfecting miR-1225-5P mimic or miR-1225-5P inhibitor into osteosarcoma cell lines. Furthermore, cell cycle analysis was performed to elucidate the possible mechanisms of the action of miR-1225-5P and YWHAZ in human osteosarcoma cells. The potential therapeutic effect of miR-1225-5p was tested in human osteosarcoma xenograft mouse model, by intravenous injection of miR-1225-5P into nude mice. Tumor sizes were measured and lung metastasis was counted after the mice were sacrificed. Results: The expression of miR-1225-5P was inversely correlated with the expression of YWHAZ in human osteosarcoma cell lines. Database search revealed that miR-1225-5P targeted YWHAZ 3' UTR. Transfection of miR-1225-5P mimic downregulated YWHAZ expression, which was demonstrated by real-time PCR, Western blot and luciferase assay. Over-expression of miR-1225-5P reduced human osteosarcoma cell growth, migration and invasion by downregulating YWHAZ expression. Cell growth, migration and invasion were increased by inhibiting miR-1225-5P in human osteosarcoma cells. The inhibition of cell growth, migration and invasion was rescued by over-expression of YWHAZ in osteosarcoma cells. Cell cycle analysis revealed that miR-1225-5P inhibited G1/G0 phase exit. In vivo xenograft model demonstrated that miR-1225-5P inhibited in vivo osteosarcoma tumor growth and lung metastasis. Conclusion: Our findings suggested that miR-1225-5P inhibits osteosarcoma cell growth in vitro and tumor growth in vivo by targeting YWHAZ. This study suggested that miR1225-5P can serve as a potential therapeutic method for treating osteosarcoma.

To explore the expression and the functions of SRPK1 in osteosarcoma, we retrieved transcription profiling dataset by array of human bone specimens from patients with osteosarcoma from ArrayExpress (accession E-MEXP-3628) and from Gene Expression Omnibus (accession GSE16102) and analyzed expression level of SRPK1 and prognostic value in human osteosarcoma. Then we examined the effect of differential SRPK1 expression levels on the progression of osteosarcoma, including cell proliferation, cell cycle, apoptosis, and investigated its underlying molecular mechanism using in vitro osteosarcoma cell lines and in vivo nude mouse xenograft models. High expression level of SRPK1 was found in human osteosarcoma tissues and cell lines as compared to the normal bone tissues and osteoblast cells, and predicted poor prognosis of human osteosarcoma. Overexpression of SRPK1 in osteosarcoma U2OS cells led to cell proliferation but inhibition of apoptosis. In contrast, knockdown of SRPK1 in HOS cells impeded cell viability and induction of apoptosis. Moreover, silencing SRPK1 inhibited osteosarcoma tumor growth in nude mice. Mechanistic studies revealed that SRPK1 promoted cell cycle transition in osteosarcoma cells and activation of NF-κB is required for SRPK1 expression and its pro-survival signaling. SRPK1 promoted human osteosarcoma cell proliferation and tumor growth by regulating NF-κB signaling pathway.

Abstract Background Osteosarcoma is the most common primary bone tumor. The survival rate of osteosarcoma patients has not significantly increased in the past decades. Uncovering the mechanisms of malignancy, progression, and metastasis will shed light on the development of new therapeutic targets and treatment for osteosarcoma. Aim The aim of this study is to identify potential osteosarcoma biomarker and/or therapeutic targets by using integrated bioinformatics analysis. Methods and results We utilized existing gene expression datasets to identify differential expressed genes (DEGs) that could serve as osteosarcoma biomarkers or even as therapeutic targets. We found 48 DEGs were overlapped in three datasets. Among these 48 DEGs, PSMD14 was on the top of the up‐regulated gene list. We further found that higher PSMD14 expression was correlated with higher risk group (younger age group, ≤20.83 years of age), metastasis within 5 years and higher grade of tumor. Higher PSMD14 expression in osteosarcoma had positive correlation with higher infiltration of CD8+ T cells, neutrophils and myeloid dendritic cells. Kaplan‐Myer survival data further revealed that higher expression of PSMD14 predicted significantly worse prognosis (p = .013). Gene set enrichment analysis was further performed for the DEGs related to PSMD14 in osteosarcoma. We found that lower PSMD14 expression group had more immune responses such as interferon γ, α responses, inflammation response etc. However, the higher PSMD14 expression group had more cell proliferation‐related biological processes, such as G2M checkpoints and Myc targets. Through establishing protein–protein interaction networks using PSMD14 related DEGs, we identified 10 hub genes that were all ribosomal proteins. These hub genes may play roles in osteosarcoma tumorigenesis, progression and/or metastasis. Conclusion We identified PSMD14 gene as a possible osteosarcoma biomarker, and/or a possible therapeutic target.

自主设计实验对学生科研能力、创新能力等综合素质及能力的培养具有重要作用,然而,在教学实践中由于受到各种因素的限制,影响了教学效果.文章从专题讲座、自主选题、开题报告、实验操作、论文撰写及对教师的要求等方面总结吉林大学药理学自主设计实验的实践经验,分析存在的问题,提出解决思路,为更好地开展药理学自主设计实验提供参考.

药理学作为沟通基础医学与临床医学的桥梁学科,在医学知识的学习过程中起着承前启后的重要作用.在药理学教学过程中,采用病例讨论式教学方法,旨在提高学生的学习兴趣,培养学生分析问题、解决问题的能力,提高教学效果.

CPT-11 (irinotecan) is a derivative of camptothecin which is a natural product derived from the Chinese tree Camptotheca acuminta and widely used in antitumor therapy. Here, the in vitro anti-tumor activity and associated mechanisms of a novel derivative of camptothecin, ZBH-1205, were investigated in a panel of 9 human tumor cell lines, as well as in HEK 293 and SK-OV-3/DPP, a multi-drug resistant (MDR) cell line, and compared to CPT-11 and 7-ethyl-10-hydroxy-camptothecin (SN38). Comparisons between the different compounds were made on the basis of IC50 values as determined by the MTT assay, and flow cytometry was used to evaluate cell cycle progression, apoptosis, and the levels of pro- and active caspase-3 among different treatment groups. Interaction between the molecules and topoisomerase-1 (Topo-1)-DNA complexes was detected by a DNA relaxation assay. Our results demonstrated that IC50 values for ZBH-1205 ranged from 0.0009 μmol/L to 2.5671 μmol/L, which were consistently lower than IC50 values of CPT-11 or SN38 in the panel of cell lines, including SK-OV-3/DPP. Furthermore, ZBH-1205 was more effective than CPT-11 or SN38 at stabilizing Topo-1-DNA complexes and inducing tumor cell apoptosis. Therefore, ZBH-1205 is a promising chemotherapeutic agent to be further assessed in large-scale clinical trials.

目的 利用Shotgun 方法分析吉林双阳梅花鹿生长期鹿茸蛋白组分,完善鹿茸蛋白质表达谱的分析.方法 先将新鲜鹿茸高速匀浆取上清;硫酸铵沉淀获取鹿茸总蛋白粗提物,DEAE纯化获取蛋白组分I;行溶液内酶解、毛细管高效液相色谱方法/质谱 shotgun分析,所得数据按照IPI号对蛋白进行GO功能在线分析.结果 鹿茸蛋白组分I共有38个蛋白质,注释蛋白34个;其中,高丰度蛋白质8个;参与生物过程的蛋白质占鹿茸蛋白组分I总量的20.5% ;参与细胞成分的蛋白质占总量的38.2%;参与分子功能的占总量的44.1%.鉴定出的蛋白质细胞内外分布平衡,已知最多的蛋白质为具有结合功能的蛋白质.结论 Shotgun方法可有效分离和分析鹿茸蛋白质组分,为鹿茸蛋白组学的研究提供了基础理论数据.

A capillary electrophoresis method coupled with electrochemiluminescence detection for the analysis of vinorelbine (VNB) in the urine of tumor patients was established in this research. Complete determination of VNB was achieved in 8 min using a background electrolyte of 50 mmol/L phosphate buffer (pH 9.0) and a separation voltage of 15 kV. The calibration curves showed a linear range from 2.8 × 10–10 to 1.6 × 10–8 mol/L. The relative standard derivation for VNB was below 3.4%. The linear relationships were good and the correlation factor of VNB exceeded 0.985. The detection limits were 1.0 × 10–11 mol/L under the optimal conditions. The developed method was applied to the sensitive determination of VNB in the urine of tumor patients.

目的:黄芪注射液对阿霉素所致心肌病理改变和超微结构的影响.方法:60只雄性Balb/c小鼠分为黄芪注射液干预组,阿霉素组和正常对照组(n=20),观察小鼠的一般情况,于14 d观察心肌病理及超微结构改变,计算心肌病理组织学积分.结果:阿霉素组心肌超微结构改变重于黄芪注射液干预组,黄芪注射液干预组与模型组比较病理积分下降明显(P＜0.01).结论:黄芪注射液有效地维护其心肌纤维及膜系统的稳定性,对阿霉素所致的心肌损伤具有保护作用.

自2007年起,吉林大学白求恩医学院药理学系开始全英讲授海外留学生中本科生和硕士研究生的药理理论课和实验课。为顺应医学教育国际化趋势的需要,培养具有国际视野和标准的临床医生,采用了主讲教材为主、辅助教材为辅的方法扩充了学生的阅读量;教学方法上,采用传统教学和PBL教学相结合;2012年春季学期,白求恩医学院的各个基础学科实施了心血管授课的整合性尝试,学期期末,按照美国职考的方式对学生进行了考核,教学效果有较大提高。

目的 探讨吉林双阳梅花鹿鹿茸蛋白组分对肿瘤细胞的细胞毒性作用.方法 将新鲜鹿茸高速匀浆上清透析后,用超滤离心法获取分子量小于10 kD,大于10 kD小于30 kD,大于30 kD的3种蛋白组分;采用MTT法,检测3种蛋白组分对人肝癌SMMC-7721 细胞系及人肺腺癌 SPC-A-1细胞系的细胞毒性作用.另外,设鹿茸提取物原浓度的1/2,1/4,1/8为高,中,低3个浓度组,以0.01 ml/g浓度皮下注射昆明小鼠,连续注射14天后,取小鼠脾细胞培养(培养基含ConA0.005 mg/ml)72 h后,加入MTT(5 mg/ml)0.01 ml,继续培养4 h,测定OD值,计算淋巴细胞转化值.结果 针对SMMC-7221细胞,分子量<10 kD,>30 kD,10 kD<<30 kD的鹿茸蛋白组分中,高剂量组的抑制率均大于30%,分子量<10 kD,10 kD<<30 kD的鹿茸蛋白组分中,中高剂量组的抑制率大于30%,而且呈现剂量依赖性关系.针对SPC-A-1细胞,从所选取的高剂量至低剂量的抑制率均小于30%.鹿茸蛋白组分高、中、低剂量组的平均OD值分别为0.46±0.05、0.29±0.03和0.17±0.03;胸腺肽阳性对照组的平均OD值为0.47±0.07;生理盐水阴性对照组的平均OD值为0.12±0.04.鹿茸蛋白组分高、中、低剂量组的转化值分别为0.34、0.17和0.05;胸腺肽阳性对照组的转化值为0.35.鹿茸蛋白组分的高剂量组和胸腺肽对照组OD值比较,差异不具有显著性(P>0.05).结论 对于SPC-A-1细胞,鹿茸各蛋白组分无明显细胞毒性作用,而对于SMMC-7221细胞,各蛋白组分的高剂量、中高剂量组则呈现一定的细胞毒性作用.鹿茸蛋白组分高剂量组具有与胸腺肽注射液相近的促进淋巴细胞转化活性.

目的建立Langendorff离体心脏灌流系统,制备缺血再灌注实验模型,观察蒺藜皂苷(gross saponinsfromTribulus terrestris,GSTT)预适应对大鼠心肌缺血再灌注损伤心功能的影响。方法 56只大鼠随机分为7组,正常对照组、缺血/再灌组(I/R)、心肌缺血预适应组(IPC)、阳性药腺苷组、GSTT 200、100、50 mg/L组,缺血30 min,再灌40 min,记录给药前、再灌后5、10、15、20、30 min心率(HR)、左室收缩压(LVSP)等心功能指标,并同步记录冠脉流量(CF),同时测定心肌组织ATP水平。结果 I/R组再灌注后5、10、15、20、30 min HR、LVSP明显下降(P<0.001);IPC组,GSTT 200、100 mg/L预适应组和腺苷预适应组再灌注后5、10、15、20、30 min HR、LVSP均明显升高(P<0.05、0.01);I/R组再灌注后各时间点(5、10、15、20、30 min)CF明显减少(P<0.001);与I/R组相比,IPC组,GSTT 200、100 mg/L预适应组和腺苷预适应组再灌注后各时间点(5、10、15、20、30 min)CF均增加(P<0.05、0.01、0.001);I/R组Na+-K+ATP酶活力明显降低(P<0.001)。与I/R组相比,IPC组,GSTT 200、100 mg/L预适应组和腺苷预适应组Na+-K+ATP酶活力明显升高(P<0.05、0.01)。I/R组的Ca2+-Mg2+ATP酶活力有增高趋势,与I/R组相比,IPC组,GSTT 200、100 mg/L预适应组和腺苷预适应组Ca2+-Mg2+ATP酶活力明显升高(P<0.05、0.01)。结论 GSTT可抑制心功能低下,舒张冠状动脉,增加冠脉血流量而改善心肌的供血、供氧;GSTT还可增加心肌组织ATP酶活性,通过抑制Na+/Ca2+交换,使得细胞摄取的Ca2+减少。

Objective To study the protective effects of gross saponins from tribulus terrestris(GSTT) on ischemia-reperfusion(I/R) injury in isolated rat heart and approach its mechanism of action.Methods Fifty-six Wistar rats were randomly divided into seven groups:control group,I/R group,ischemic preconditioning(IPC) group,adenosine group,and GSTT 200,100,50 mg·L-1 groups.The isolated hearts were subjected to 30 min ischemia and then followed by 40 min reperfusion.The contents of CK,LDH,AST,MDA and activity of SOD in reperfused ischemic rat heart were detected.The histopathological changes of myocardium were observed by hematoxylin-eosin(HE) staining.Results Conpared with I/R group,the contents of CK,LDH,AST were decreased in GSTT 200 and 100 mg·L-1 groups(P0.05 and P0.01),and the contents of MDA in myocardium tissues were also decreased(P0.05 and P0.01),and the activities of SOD were increased(P0.05 and P0.01).GSTT could alleviate the pathological lesion of myocardium tissues.Conclusion GSTT has protective effect on cadiocytes injured by ischemia-reperfusion,its mechanism may be concerned with resisting oxygen free radical.

microRNA(miRNA)是近年来发现的一类长20～24nt的非编码小分子RNA,它作为一种调控因子,与siRNA有一定的相似处,在细胞的分化、增殖和凋亡、个体发育、机体代谢巾都具有重要的作用.最近的研究发现.miRNA表达与多种癌症相关,故可考虑将其作为抗肿瘤药物或者其他抗肿瘤药物的靶分子.miRNAs对癌的发生、发展及其治疗策略均有影响.肺癌为目前较为高发的恶性肿瘤,发病机制尚未完全清楚.本文就近年来关于miRNAs的研究文献,从肿瘤发生的分子水平,对miRNA与肺癌的相戈性作一综述,