Receptor-interacting protein kinase 1 (RIPK1) plays a crucial role in controlling inflammation and cell death. Its function is tightly controlled through post-translational modifications, enabling its dynamic switch between promoting cell survival and triggering cell death. Phosphorylation of RIPK1 at various sites serves as a critical mechanism for regulating its activity, exerting either activating or inhibitory effects. Perturbations in RIPK1 phosphorylation status have profound implications for the development of severe inflammatory diseases in humans. This review explores the intricate regulation of RIPK1 phosphorylation and dephosphorylation and highlights the potential of targeting RIPK1 phosphorylation as a promising therapeutic strategy for mitigating human diseases.

Cytokine release syndrome, also called cytokine storm, could cause lung tissue damage, acute respiratory distress syndrome (ARDS) and even death during SARS-CoV-2 infection. However, the underlying mechanisms of cytokine storm still remain unknown. Among these cytokines, the function of TNF-α and type I IFNs especially deserved further investigation. Here, we first found that TNF-α and IFN-β synergistically induced human airway epithelial cells BEAS-2B death. Mechanistically, the combination of TNF-α and IFN-β led to the activation of caspase-8 and caspase-3, which initiated BEAS-2B apoptosis. The activated caspase-8 and caspase-3 could further induce the cleavage and activation of gasdermin D (GSDMD) and gasdermin E (GSDME), which finally resulted in pro-inflammatory pyroptosis. The knock-down of caspase-8 and caspase-3 could effectively block the activation of GSDMD and GSDME, and then the death of BEAS-2B induced by TNF-α and IFN-β. In addition, pan-caspase inhibitor Z-VAD-FMK (ZVAD) and necrosulfonamide (NSA) could inhibit BEAS-2B death induced by TNF-α and IFN-β. Overall, our work revealed one possible mechanism that cytokine storm causes airway epithelial cells (AECs) damage and ARDS. These results indicated that blocking TNF-α and IFN-β-mediated AECs death may be a potential target to treat related viral infectious diseases, such as COVID-19.

目的 观察聚肌胞苷酸[polyinosinic-polycytidylic acid,简称为Poly(I∶C)]和脂多糖(lipopolysaccharide,LPS)预活化后的人脂肪间充质干细胞(human adipose-derived stem cells,bADMSCs)对人肺癌H460细胞生长的抑制作用.方法 利用抽脂手术遗弃脂肪组织标本制备hADMSCs,并鉴定其细胞表面分子标志、细胞增殖能力和体外分化能力;比较Poly(I∶C)、LPS预活化后的hADMSCs细胞表面标志和体外分化能力;ELISA法测定Poly(I∶C)、LPS预活化后的hADMSCs培养基上清中IFNβ的分泌情况;Western blot法分析Poly(I∶C)预活化后的hADMSCs TRAIL的表达情况;用不同浓度Poly(I∶C)预活化hADMSCs后,与不同比例的H460共培养,分析对肿瘤细胞的抑制作用.结果 光学显微镜下hADMSCs呈纤维样、鱼群样排列;hADMSCs细胞表面标志CD44、CD73、CD90、CD105、HLA-ABC的阳性率均大于95％,造血干细胞的特异性标志物CD34、CD45的阴性率均大于98％;Poly(I∶C)和LPS刺激的hADMSCs其细胞表面标志未受影响.Poly(I∶C)和LPS能促进hADMSCs的成骨分化,对成脂分化能力无明显作用.经Poly(I∶C)诱导的hADMSCs分泌的IFNβ浓度明显高于未经诱导的对照组(P＜0.05),低浓度的LPS诱导hADMSCs IFNβ表达量不高.Poly(I∶C)预处理促进了hADMSCs表达TRAIL.体外5和25 μg/mL的Poly(I∶C)预活化后的hADMSCs对H460细胞表现出细胞毒性,对H460细胞的生长有抑制作用;hADMSCs∶H460比例为1∶2时,对H460细胞的抑制效果最佳.结论 成功分离、培养出hADMSCs,其在体外低浓度的Poly(I∶C)诱导下能表达TRAIL,并对H460细胞的生长产生抑制作用.

Receptor-interacting protein kinase 1 (RIPK1) is a key regulator of inflammation and cell death. Many sites on RIPK1, including serine 25, are phosphorylated to inhibit its kinase activity and cell death. How these inhibitory phosphorylation sites are dephosphorylated is poorly understood. Using a sensitized CRISPR whole-genome knockout screen, we discover that protein phosphatase 1 regulatory subunit 3G (PPP1R3G) is required for RIPK1-dependent apoptosis and type I necroptosis. Mechanistically, PPP1R3G recruits its catalytic subunit protein phosphatase 1 gamma (PP1γ) to complex I to remove inhibitory phosphorylations of RIPK1. A PPP1R3G mutant which does not bind PP1γ fails to rescue RIPK1 activation and cell death. Furthermore, chemical prevention of RIPK1 inhibitory phosphorylations or mutation of serine 25 of RIPK1 to alanine largely restores cell death in PPP1R3G-knockout cells. Finally, Ppp1r3g −/− mice are protected from tumor necrosis factor-induced systemic inflammatory response syndrome, confirming the important role of PPP1R3G in regulating apoptosis and necroptosis in vivo.

Current studies have shown that type I or II interferon-modified mesenchymal stem cells have great potential for the application of tumor-targeted therapy, but the underlying mechanism remains largely elusive. Here, we compared the different effects of IFN-β and IFN-γ on the antitumor activity of human amniotic fluid-derived mesenchymal stem cells (AFMSCs) and revealed the potential mechanism. In detail, AFMSCs primed with IFN-β or IFN-β plus IFN-γ, not IFN-γ, inhibited the proliferation of cancer cells in an immunocompetent mouse H460 subcutaneous model, although they all inhibited the proliferation of cancer cells in an immunocompromised mouse H460 subcutaneous model. TRAIL expressed by IFN-β- or IFN-γ-primed AFMSCs specifically exerted the antitumor effect of AFMSCs. AFMSCs primed with IFN-γ highly expressed immunosuppressive molecule IDO1, but IFN-β counteracted the IFN-γ-initiated IDO1 expression. 1-MT (IDO1 inhibitor) decreased TRAIL, but increased IDO1 expression in AFMSCs primed with interferon. As a result, AFMSCs primed with IFN-β or IFN-γ had the antitumor activity, and 1-MT failed to enhance the antitumor effect of IFN-γ-primed AFMSC in vitro and in the immunocompromised mouse H460 subcutaneous model. Furthermore, the expression of TRAIL in AFMSCs was upregulated by apoptotic cancer cells and this positive feedback intensified the antitumor effects of IFNs-primed AFMSCs. The different target gene expression profiles of AFMSCs regulated by IFN-β and IFN-γ determined the different antitumor effects of IFN-β- and IFN-γ-primed AFMSCs on tumor cells. Our finding may help to explore a clinical strategy for cancer intervention by understanding the antitumor mechanisms of MSCs and interferon.

广州医科大学金域检验学院作为广东省第一家医学检验技术专业试点学院,采用了高校、企业、医院三方联合优势资源办学的模式,而正是因为这种模式的存在,也凸显了其特有的特点:1.教师来源较分散;2.实习基地较多;3.各部门之间发展水平不均衡.这就要求教学管理人员要想尽一切办法来提高工作效率,保证工作质量.通过建设金域检验学院网络信息资源库,提升数据来源的准确性,加快数据更新的速度,从而加快教学改革的进程,提高教学管理的效率,促进教学水平的提高.

目的 利用慢病毒转染的方法将人干扰素β基因(hIFN-β)、人干扰素γ基因(hIFN-γ)分别导入人羊水来源的间充质干细胞(AFMSCs),以期获得稳定表达hIFN-β、hIFN-γ的AFMSCs.方法 产前诊断获得羊水标本,培养出AFMSCs,对其增殖能力、细胞表面分子和体外分化能力进行鉴定.用携带hIFN-β、hIFN-γ基因的重组慢病毒颗粒感染AFMSCs,并用Elisa法对培养基上清中hIFN-β、hIFN-γ的分泌情况进行鉴定.结果 人羊水间充质干细胞呈成纤维样形态,不表达CD34,高表达CD44、CD105和HLA-ABC,能稳定传代培养,具有成骨、成脂分化能力.ELISA结果显示hIFN-β、hIFN-γ修饰的AFMSCs能稳定表达hIFN-β、hIFN-γ,其表达量分别达1 830 pg/mL和65.33 pg/mL.结论 利用慢病毒转染的方法成功制备了hIFN-β、hIFN-γ修饰的人羊水间充质干细胞.

Objective To investigate the effect of the cyclin-dependent kinase inhibitor SNS-032 on the growth of taxol-resistant MCF-7/TAX breast cancer cells. Methods The resistance situation of MCF-7/TAX cells to paclitaxel and the effect of SNS-032 on the growth of taxol-resistant MCF-7/TAX cells were evaluated by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide) assay. The effect of SNS-032 on apoptosis in MCF-7/TAX cells was analyzed by Annexin V-FITC/PI staining. Results The resistance index of MCF-7/TAX cells to paclitaxel was 19.39. After treatment with SNS-032 for 48 h, MCF-7/TAX cells displayed markedly inhibited growth in a dose-de-pendent manner, and calculated IC50 was 164.5 nmol/L. SNS-032 decreased the resistance index of MCF-7/TAX from 19.39 to 0.89. The numbers of apoptotic MCF-7/TAX cells, as revealed by Annexin V binding, significantly increased upon SNS-032 treatment (P<0.05). Conclusion The results demonstrate that SNS-032 can induce apoptosis in MCF-7/TAX cells and consequently display potent cytocidal effect on taxol-resistant MCF-7/TAX breast cancer cells.

Objective To investigate the effect of JNJ-7706621 on the growth of adriamycin-resistant MCF-7/ADR breast cancer cells.Methods The effect of JNJ-7706621 on the growth of adriamycin-resistant MCF-7/ADR cells was determined by MTT assay.The effect of JNJ-7706621 on apoptosis in MCF-7/ADR breast cancer cells was evaluated by TUNEL staining, and the activity of Caspase-8 and Caspase-9 was quantified by enzymatic activity assay.The effect of JNJ-7706621 on cell cycle progression was analyzed by flow cytometry analysis.Results After treatment with JNJ-7706621 for 48 h, MCF-7/ADR cells displayed markedly inhibited growth in a dose-dependent manner, and calculated IC50 was 0.83 μmol/L.The numbers of apoptotic MCF-7/ADR cells, as revealed by TUNEL staining, significantly increased after JNJ-7706621 treatment (P<0.05).The enzymatic activity of Caspase-8 and Caspase-9 markedly increased after JNJ-7706621 treatment (P<0.05).JNJ-7706621 treatment caused an increase of cells in G2/M phase from (7.1±1.3)% to (23.8±3.1)% (P<0.05).Conclusion These data demonstrates that JNJ-7706621 induces apoptosis through both intrinsic and extrinsic apoptotic pathways, in the meanwhile, JNJ-7706621 induces G2/M cell cycle arrest in MCF-7/ADR cells, and consequently suppress the growth of adriamycin-resistant MCF-7/ADR breast cancer cells.

Objective To construct the prokaryotic expression vector of TAT?Apoptin,express and purify TAT?Apoptin,and detect its inhibitory effect on tumor cells. Methods The TAT?Apoptin sequence was amplified by PCR,and cloned into the multiple clone sites of plasmid pET?28b(+),and then the ligation product was transformed into E.coli DH5α. After identified by double enzyme?cutting and DNA sequencing,the recombinant plasmid was transformed into E.coli BL21 and the target protein was induced to express by IPTG. The TAT?Apoptin was purified by Ni?NTA affinity chromatography. The anti?tumor effect of TAT?Apoptin on three kinds of tumor cells was evaluated by morphology observation and CCK8 assay. Results The recombinant plasmid pET?28b?Apoptin was constructed successfully. The TAT?Apoptin was induced to express by IPTG and identified by SDS?PAGE. The purified TAT?Apoptin could promote the apoptosis of tumor cells,such as A549,MDA?MB?231 and SKB?R3. Conclusions The prokaryotic expression vector of TAT?Apoptin was constructed. The TAT?Apoptin had effect on promoting apoptosis of tumor cells.

目的 研究雷公藤红素对人阿霉素耐药MCF-7/ADR乳腺癌细胞生长的作用.方法 采用MTT试验检测MCF-7/ADR细胞对阿霉素的耐药情况以及雷公藤红素对MCF-7/ADR细胞生长的影响;采用Annexin V-FITC/PI双染试验分析雷公藤红素对MCF-7/ADR耐药细胞凋亡的诱导作用;应用流式细胞周期分析检测雷公藤红素对MCF-7/ADR耐药细胞周期的影响.结果 MCF-7/ADR细胞对阿霉素耐药指数达14.54;而雷公藤红素能有效抑制阿霉素耐药细胞MCF-7/ADR的生长,并呈现浓度依赖性,作用48 h的IC50是1.04 μmol/L,MCF-7/ADR对雷公藤红素的耐药指数仅为0.87.2 μmol/L雷公藤红素作用8 h后,Annexin V-FITC染色阳性的MCF-7/ADR细胞比例较对照显著升高,差异有统计学意义(P<0.05).在1 μmol/L雷公藤红素作用24 h后,G1期细胞比例由对照(59.22±3.78)%升高至(77.44±4.21)%,而S期细胞比例由对照(37.51±2.91)%降至(19.65±2.25)%,差异有统计学意义(P<0.05).结论 雷公藤红素能激活MCF-7/ADR细胞凋亡的发生,并诱导MCF-7/ADR发生 G1/S细胞周期阻滞,从而对阿霉素耐药MCF-7/ADR细胞的生长发挥高效抑制作用.

Objective:To analyze chromosomal abnormalities of chorionic villus cell in patients with early spontaneous abortion.Methods:The chorionic villus tissues of women with early spontaneous abortion were digested by collagenase as well as trypsin and cultured in CO2 incubator for 7 days.The karyotype was observed by Giemsa staining method after adherent cells were treated with colchicines.The chromosome number,structural abnormalities,sex ratio and age of the pregnant women were analyzed.Results:In 131 cases of villus samples,129 specimens were successfully cultured.Among them,there were 61 cases of abnormal karyotypes;trisomy in abnormal karyotype was 52.4％;the monomer was 18％;the triploid was 16.4％.The majority of aborted embryos were female,but the sex difference was no statistical significance in normal karyotypes and abnormal karyotypes.The proportion of chromosome abnormalities in the specimens obtained from women over 35 years was significantly higher than that of women under 35 years(P ＜ 0.05).Conclusion..Chorionic villus karyotype analysis can be used to determine the cause of miscarriage in early spontaneous abortion,which is instructive to the successful pregnancy of the pregnant woman.

目的 研究雷公藤红素对紫杉醇耐药乳腺癌细胞生长的作用及其机制.方法 用MTIT法检测雷公藤红素对紫杉醇耐药MCF-7/TAX细胞生长的影响;Annexin V-FITC/PI染色检测雷公藤红素对MCF-7/TAX凋亡的诱导作用;Caspase酶活性测定分析雷公藤红素作用后,MCF-7/TAX细胞中Caspase-8、-9活化程度.结果 MCF-7/TAX对紫杉醇耐药指数达19.39;而雷公藤红素对MCF-7/TAX细胞的生长有高效抑制作用,且呈现明显的剂量依赖性,对其作用48 h的IC50为0.92 μmol/L,MCF-7/TAX对雷公藤红素的敏感性高于MCF-7,耐药指数仅为0.77.雷公藤红素作用后,Annexin V染色阳性的MCF-7/TAX细胞较对照明显升高.Caspase酶活性测定结果显示,雷公藤红素作用24 h后,MCF-7/TAX细胞中Caspase-8、-9酶活性均显著升高.结论 雷公藤红素能同时激活内外源性凋亡途径,诱导MCF-7/TAX细胞凋亡的发生,从而高效抑制紫杉醇耐药MCF-7/TAX乳腺癌细胞的生长.

Objective To explore the relationship between prenatal diagnosis indications and fetal chromosomal aberrations , and the security of amniocentesis. Methods The amniotic fluid cells were sampled by amniocentesis and cultured in 572 high-risk pregnant women from January 2012 to August 2015. The chromosomal karyotypes were examined by G-banding. Results The success rate of the first amniotic fluid cells culture reached 99.83%. In all the 572 valid samples , there were 20 cases of chromosomal aberrations and the abnormal rate was 3.50%, including 17 of numeric aberrations and 3 of structural aberrations. There were 7 cases of chromosomal aberrations in all the 299 elderly parturient in high-risk indications and the abnormal rate was 2.34%, and there were 13 cases of chromosomal aberrations in all the 273 non-elderly parturient and the abnormal rate was 4.76%. Conclusions (1)It is necessary to further diagnose in pregnant women with high-risk factors , including high age , abnormal screening and ultrasonic findings , and history of abnormal gestation and birth. (2)The chromosomal karyotype examination of amniotic fluid cells in high-risk pregnant women is one of the effective prenatal diagnosis methods in high security and accuracy , with which it can reduce the incidence of birth defects and the burden of family and society , and improve the quality of the population.

Objective To evaluate the role of peripheral blood chromosomal karyotype analysis in disease diag-nosis of chromosomal disorder. Methods The cytogenetic karyotype of peripheral blood derived from 780 genetic counseling cases were analyzed during January 2013 to August 2015. Results There were 30 cases of chromosomal ab-errations to be detected and the abnormal rate was 3.85%, including 12 cases of numeric aberrations and 18 cases of structural aberrations. In neonatal disorder group, developmental disorder group and infertile group, the main type of chromosomal aberrations was the numeric abnormality, and it was mainly structural abnormality in the abnormal gesta-tion and birth group. Conclusion The detection of the peripheral blood chromosomal karyotype may play an important role in the diagnosis of neonatal disorder, developmental disorder, infertility and abnormal gestation and birth.

Objective To investigate the effect of pristimerin on the growth and apoptosis of MCF-7 breast cancer cells and to explore the underlying mechanism.Methods The effect of pristimerin on the growth and of apoptosis MCF-7 cells were analyzed by MTT assay and Annexin Ⅴ-FITC/PI staining,TUNEL staining,respectively.The ex pression of Bcl-2 and Bax was determined by western blotting analysis.Results Pristimerin exhibited a marked inhibi tion on the survival of MCF-7 cells,and the half maximal inhibitory concentration (IC50) value was 0.59 μmol/L.The numbers of apoptotic MCF-7 cells,as revealed by Annexin Ⅴ binding and TUNEL assay,increased upon pristimerin treatment.Pristimerin treatment resulted in a decrease of Bcl-2 and an increase of Bax expression.Conclusion Our results demonstrated that pristimerin suppressed the proliferation of MCF-7 cells,and that these effects occurred through increasing apoptosis by regulation of apoptotic effectors.

目的:研究扁蒴藤素对紫杉醇耐药乳腺癌细胞MCF-7/TAX生长的作用.方法:以乳腺癌细胞株MCF-7为对照,同时制作紫杉醇耐药乳腺癌细胞MCF-7/TAX;在两组细胞中分别给与7种浓度(0.25、0.5、1、2.5、5、10和20 μmol/L)扁蒴藤素或紫杉醇培养48 h,用MTT法检测药物对MCF-7和MCF-7/TAX细胞作用的半数抑制浓度(IC50)值,并计算MCFG/TAX细胞对紫杉醇或扁蒴藤素的耐药指数(RI);同时用Annexin V-FITC/PI染色观察扁蒴藤素作用后MCF-7/TAX凋亡细胞比例.结果:MCF-7/TAX对紫杉醇RI达19.39,而扁蒴藤素对MCF-7/TAX细胞的生长有高效抑制作用,且呈现明显的剂量依赖性,对其作用48 h的IC5o为0.52 μmol/L,MCF-7/TAX对扁蒴藤素的RI仅为0.88;扁蒴藤素作用后,Annexin V染色阳性的MCF-7/TAX细胞比例较对照明显升高.结论:扁蒴藤素能抵抗紫杉醇耐药乳腺癌细胞的耐药性,激活MCF-7/TAX细胞凋亡,对其生长发挥高效抑制作用.

Human mesenchymal stem cells (MSCs) represent a novel carrier for gene therapy and apoptin is a potential tumor-selective apoptosis-inducing protein. In the present study, the anti-tumoral effect of MSCs modified with apoptin against lung carcinoma was evaluated. Apoptin protein was expressed in a prokaryotic expression system and purified by affinity chromatography. Subsequently, anti-apoptin antibody was prepared by immunizing BALB/c mice with purified apoptin protein. Human MSCs were isolated, amplified and transduced with lentiviral vectors encoding full-length apoptin, in which the secretory signal and protein transduction sequence were added into the amino terminus to assist apoptin in entering into target cells. The differentiation and apoptin expression of apoptin-modified MSCs were confirmed. Subsequently, the anti-tumor effect of apoptin-modified MSCs was measured in vitro and in vivo. Following modification with apoptin, MSCs retained their differentiation capacity, and successfully synthesized and secreted apoptin, which entered target cells and selectively induced lung cancer cell apoptosis through activating caspase-3. The percentage of tumor cells with activated caspase-3 in the apoptin-modified MSCs group was markedly higher than that in the MSCs group (16.5±2.9% at 24 h and 27.3±2.0% at 48 h vs. 3.4±1.1% at 24 h and 2.2±0.6% at 48 h). When injected into nude mice, apoptin-modified MSCs inhibited the growth of lung carcinoma compared with that in the control groups (0.14±0.02 g vs. 0.21±0.04 g vs. 0.31±0.05 g, P<0.05). The results of the present study provided preclinical support of apoptin-based cancer therapy with MSCs as cellular vehicles.

为了适应新的人才培养需求和学科发展趋势，广州医科大学对目前的临床免疫学检验实验课程进行了合理调整和改革，并进行了初步实践。根据教学目标和学生接受能力存在个体化差异的特点，遵循进阶式教学规律，按照模块化教学模式对临床免疫学检验实验课程内容进行重新设计和优化，在保证必要基础性实验的前提下，强化综合性实验内容，并针对性地设置研究性实验，与此同时相应配套改革现有实验课程成绩考核方式。通过以上各个环节的紧密配合和衔接，力争在培养创新型医学检验技术人才方面作出一些有益探索和尝试。

As one tumor marker of HCC, Golgi Protein 73 (GP73) is given more promise in the early diagnosis of HCC, and aptamers have been developed to compete with antibodies as biorecognition probes in different detection system. In this study, we utilized GP73 to screen specific ssDNA aptamers by SELEX technique. First, GP73 proteins were expressed and purified by prokaryotic expression system and Nickle ion affinity chromatography, respectively. At the same time, the immunogenicity of purified GP73 was confirmed by Western blotting. The enriched ssDNA library with high binding capacity for GP73 was obtained after ten rounds of SELEX. Then, thirty ssDNA aptamers were sequenced, in which two ssDNA aptamers with identical DNA sequence were confirmed, based on the alignment results, and designated as A10-2. Furthermore, the specific antibody could block the binding of A10-2 to GP73, and the specific binding of A10-2 to GP73 was also supported by the observation that several tumor cell lines exhibited variable expression level of GP73. Significantly, the identified aptamer A10-2 could distinguish normal and cancerous liver tissues. So, our results indicate that the aptamer A10-2 might be developed into one molecular probe to detect HCC from normal liver specimens.