Research progress on ribosomal proteins and their functions in diseases / 药学实践杂志

Ribosomal proteins (RP) are important components of ribosomes and play key roles in ribosome biogenesis and protein translation. In addition, ribosomal proteins also possess many extra-ribosomal functions, such as regulation of gene expression, cell proliferation, differentiation, apoptosis, DNA repair, and many other cellular processes. The dysfunction of RP is closely related to the occurrence and development of various diseases including blood, metabolism, cardiovascular diseases and tumors, and RP might become potential therapeutic targets for a variety of diseases. The research progress on RP, including the basic functions of RP, extra-ribosomal properties, and the connections with human diseases were reviewed and their potential as biomarkers and therapeutic targets in diseases were discussed in this paper.

Effects of specific knockdown of ribosomal protein S5 in hepatic stellate cells on liver fibrosis / 药学实践杂志

Objective To observe the effect of specific knockdown of hepatic stellate cells (HSC) ribosomal protein S5 (RPS5) on liver fibrosis in rats. Methods The glial fibrillary acidic protein (GFAP) promoter-driven RPS5 shRNA adenovirus was established, and AdGFa2-shRPS5 and its control AdGFa2 shNC were used to transfect primary rat HSCs and hepatocytes, respectively. RPS5 was determined by Western-blot and Real Time PCR, α-SMA and type I collagen expression; the rat liver fibrosis model was established by dimethyl nitrosamine (DMN) and bile duct ligation (BDL), and intrahepatic HSC was specifically knocked down by tail vein injection of adenovirus of RPS5 levels. The pathological changes of liver tissue sections were analyzed by HE staining; the content of hydroxyproline, sections of Sirius red and Masson staining were used to evaluate collagen deposition; immunohistochemical staining was used to detect the expression of α-SMA and RPS5. Results AdGFa2-shRPS5 specifically knocked down the expression level of RPS5 in HSC and increased the expression of α-SMA and type I collagen in vitro. The in vivo results showed that in two animal models of chronic liver injury, specific knockdown of RPS5 expression in HSCs promoted HSC activation, increased the deposition of extracellular matrix, and promoted liver fibrosis. Conclusion RPS5 is essential for HSC activation and liver fibrosis, which could be a potential target for the treatment of liver fibrosis.

Knockout of ribosomal genes bS22 and bL37 increases the sensitivity of mycobacteria to antibiotics / 生物工程学报

In recent years, two novel proteins in the ribosomes of mycobacteria have been discovered by cryo-electron microscopy. The protein bS22 is located near the decoding center of the 30S subunit, and the protein bL37 is located near the peptidyl transferase center of the 50S subunit. Since these two proteins bind to conserved regions of the ribosome targeted by antibiotics, it is speculated that they might affect the binding of related drugs to these targets. Therefore, we knocked out the genes encoding these two proteins in wild-type Mycolicibacterium smegmatis mc2155 through homologous recombination, and then determined the growth curves of these mutants and their sensitivity to related antibiotics. The results showed that compared with the wild-type strain, the growth rate of these two mutants did not change significantly. However, mutant ΔbS22 showed increased sensitivity to capreomycin, kanamycin, amikacin, streptomycin, gentamicin, paromomycin, and hygromycin B, while mutant ΔbL37 showed increased sensitivity to linezolid. These changes in antibiotics sensitivity were restored by gene complementation. This study hints at the possibility of using ribosomal proteins bS22 and bL37 as targets for drug design.

MicroRNA-4699-3p inhibits the proliferation and migration of ovarian cancer cells by regulating the expression of mitochondrial ribosomal protein S23 / 中国医师杂志

Objective:To investigate the expression of microRNA (miRNA, miR)-4699-3p in ovarian cancer cell lines, and observe its ability to regulate the expression of mitochondrial ribosomal protein S23 (MRPS23) and its effect on the migration and proliferation of ovarian cancer cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-4699-3p in ovarian cancer cell lines (OVCAR-3, SKOV-3, HO-8910, OC3, A2780) and normal ovarian epithelial cells (IOSE80). The liposome transfection method was used to transfect miR-4699-3p mimic or negative control miR-NC to the cell line with the lowest miR-4699-3p expression, which was defined as the experimental group and the control group. qRT-PCR was used to verify transfection efficiency. Bioinformatics technology predicted the candidate target gene of miR-4699-3p, and the dual luciferase reporter gene experiment identified its ability to regulate the target gene. qRT-PCR and Western blot were used to detect the expression of target genes at the mRNA and protein levels. Cell counting method (CCK-8) and transwell migration experiment were used to detect the effect of miR-4699-3p on the proliferation and migration of ovarian cancer cells.Results:The expression of miR-4699-3p in ovarian cancer cell lines was significantly lower than that of normal ovarian epithelial cells ( P<0.05), and the lowest expression in OVCAR-3 cells ( P<0.01). After transfection of miR-4699-3p, the expression of miR-4699-3p in OVCAR-3 cells in the experimental group was significantly increased ( P<0.01), which proved that the transfection was successful. Bioinformatics technology predicted that the candidate target gene of miR-4699-3p may be MRPS23. The dual luciferase reporter gene experiment confirmed that miR-4699-3p can directly target the 3′-UTR of MRPS23 gene mRNA ( P<0.01). Compared with the OVCAR-3 cells in the control group, the mRNA and protein expression levels of the MRPS23 gene were significantly reduced, while the expressions of Twist, Slug, CDK6 and Cyclin D3 were significantly decreased ( P<0.01) in the experimental group after up-regulating miR-4699-3p expression ( P<0.01). After up-regulating miR-4699-3p expression, the proliferation ability of OVCAR-3 cells decreased ( P<0.05) and the migration ability of OVCAR-3 cells was reduced ( P<0.01). Conclusions:miR-4699-3p is under-expressed in ovarian cancer cell lines. Up-regulation of miR-4699-3p expression in OVCAR-3 cells can inhibit ovarian cancer cell proliferation and migration by interfering with MRPS23 gene expression.

Kaempferol promotes osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress via the mTORC1 signaling pathway / 口腔疾病防治

Objective @#To investigate the activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway molecules during the process by which kaempferol (Kae) promotes osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMCs) under cyclic and uniaxial tension.@*Methods @#BMMCs isolated and cultured in vitro were subjected to uniaxial dynamic tension with a 10% shape variable. The appropriate concentration of Kae was selected by cytotoxicity testing. The endogenous mTOR signal was inhibited by pp242. Four hours after traction, alkaline phosphatase (ALP) and osteocalcin (OCN) were detected by chemical colorimetry and ELISA, and the relative concentration of intracellular calcium was detected by flow cytometry. Phosphorylation of mTOR, 4E/BP1, and ribosomal protein S6 kinases (S6K), which are the main molecules of the endogenous mTORC1 signaling pathway, and expression of osteogenic transcription factors (Runx2 and Osterix) were detected by western blotting (WB), and mRNA expression levels of the above factors were detected by qRT-PCR.@*Results @# The cytotoxicity test showed that 10 μmol/L Kae had little inhibitory effect on cell proliferation but had the strongest osteogenic ability. Four hours after stretching, Kae effectively promoted the osteogenic differentiation of BMMCs. The expression of ALP was （153.04 ± 18.72） U/mg, the expression of OCN was （1.64 ± 0.25） U. The mRNA and protein levels of Runx2 and Osterix were upregulated, and the intracellular calcium content was decreased. The mRNA and protein phosphorylation of mTOR and S6K was upregulated, and the opposite effect was observed with 4E/BP1. After pp242 was added to inhibit mTOR signaling, mTOR and S6K mRNA and protein phosphorylation were downregulated, but 4E/BP1 mRNA and protein phosphorylation was upregulated. The osteogenic differentiation of BMMCs was also significantly inhibited, mRNA and protein expression of Runx2 and Osterix were significantly downregulated, ALP and OCN expression were downregulated, and intracellular calcium content was increased. @* Conclusion@#Kae promotes osteogenic differentiation of mouse BMMCs under uniaxial dynamic tension through the mTORC1 signaling pathway.

Predicting the Benefit of Adjuvant Aromatase Inhibitor Therapy in Postmenopausal Breast Cancer Patients with Phosphorylated S6K1 Expression Status / 한국유방암학회지

PURPOSE: Phosphorylated ribosomal S6 kinase 1 (pS6K1) is a major downstream regulator of the mammalian target of rapamycin (mTOR) pathway. Recent studies have addressed the role of S6K1 in adipogenesis. pS6K1 may affect the outcome of estrogen depletion therapy in patients with hormone-sensitive breast cancer due to its association with adipogenesis and increased local estrogen levels. This study aimed to investigate the potential of pS6K1 as a predictive marker of adjuvant aromatase inhibitor (AI) therapy outcome in postmenopausal or ovarian function-suppressed patients with hormone-sensitive breast cancer.METHODS: Medical records were retrospectively reviewed in postmenopausal or ovarian function-suppressed patients with estrogen receptor-positive and node-positive primary breast cancer. pS6K1 expression status was scored on a scale from 0 (negative) to 3+ (positive) based on immunohistochemical analysis.RESULTS: A total of 428 patients were eligible. The median follow-up duration was 44 months (range, 1–90). In patients with positive pS6K1 expression, AIs significantly improved disease-free survival (DFS) compared to selective estrogen receptor modulators (SERMs) (5 year-DFS: 83.5% vs. 50.7%, p = 0.016). However, there was no benefit of AIs on DFS in the pS6K1 negative group (5 year-DFS 87.6% vs. 91.4%, p = 0.630). On multivariate analysis, AI therapy remained a significant predictor for DFS in the pS6K1 positive group (hazard ratio, 0.39; 95% confidence interval, 0.16–0.96; p = 0.041). pS6K1 was more effective in predicting the benefit of AI therapy in patients with ages < 50 (p = 0.021) compared to those with ages ≥ 50 (p = 0.188).CONCLUSION: pS6K1 expression may predict AI therapy outcomes and serve as a potential predictive marker for adjuvant endocrine therapy in postmenopausal and ovarian function-suppressed patients with hormone-sensitive breast cancer. AIs may be more effective in patients with pS6K1 positive tumors, while SERM could be considered an alternative option for patients with pS6K1 negative tumors.

Expression of phosphoinositide 3-kinase, mammalian target of rapamycin, 70-kDa ribosomal protein S6 kinase, and interferon-α in umbilical cord blood plasmacytoid dendritic cells in parturients in the immune tolerance stage of hepatitis B virus infection versus normal parturients / 临床肝胆病杂志

ObjectiveTo investigate the expression of phosphoinositide 3-kinase (PI3K), mammalian target of rapamycin (mTOR), 70-kDa ribosomal protein S6 kinase (p70S6K), and interferon-α (IFNα) in umbilical cord blood plasmacytoid dendritic cells (pDCs) in parturients in the immune tolerance stage of hepatitis B virus (HBV) infection versus normal parturients. MethodsA total of 20 parturients in the immune tolerance stage of HBV infection and 10 normal parturients who were hospitalized in Inpatient Department of Obstetrics in The First Affiliated Hospital of Hunan University of Chinese Medicine from October 2017 to January 2020 were enrolled as hepatitis B group and normal group, respectively. Umbilical cord blood pDCs were isolated and cultured, and CpG-A was added on day 7. The cells and the supernatant were collected after 24 hours; real-time PCR was used to measure the mRNA expression of PI3K, mTOR, and p70S6K, Western Blot was used to measure the protein expression of PI3K, mTOR, and p70S6K, and ELISA was used to measure the level of IFNα in the supernatant of pDCs. The two-independent-samples t-test was used for comparison of continuous data between the two groups. ResultsCompared with the normal group, the hepatitis B group had significantly lower mRNA and protein expression of PI3K, mTOR, and p70S6K in umbilical cord blood pDCs (mRNA expression: t=-81.04, -63.07, and -34.55, all P＜0.001; protein expression: t=-8.13, -7.75, and -6.71, all P＜0.001). The hepatitis B group had significantly lower expression of IFNα in the supernatant of umbilical cord blood pDCs than the normal group (t=-15.88, P＜0.05). ConclusionParturients in the immune tolerance stage of HBV infection have reductions in the mRNA and protein expression of PI3K, mTOR, and p70S6K and the level of IFNα in umbilical cord blood pDCs, suggesting that pDC function is inhibited.

The role of the mTORC1 signaling pathway during osteogenic differentiation of mouse bone marrow mesenchymal cells under tension stress / 口腔疾病防治

Objective@# To investigate the expression of the mTORC1 signaling pathway during the osteogenic differentiation of mouse bone marrow mesenchymal cells (BMMSCs) under cyclic uniaxial tension and explore its possible role.@*Methods @# The BMMSCs of mice were affected by uniaxial dynamic tensile force. Western blot was used to detect the expression changes of major molecules (mTOR, Raptor, S6K) in the endogenous mTORC1 signaling pathway at 0, 1, 2, 4, and 8 hours after stretching. Chemical colorimetry, ELISA and PCR were used to detect alkaline phosphatase (ALP), osteocalcin (OCN) and Runx2 mRNA, respectively. Then, inhibition, activation and control groups were established by administration of the drugs PP242, MHY1485 and PBS, respectively. Two hours after the stress, the expression of S6K was detected by western blot, and the expression of the osteogenic signal was continuously detected by the above methods.@*Results @#Western blot analysis showed that the main molecules of the mTORC1 signaling pathway were all expressed within 8 hours after traction, and the highest expression was 2 hours after the stress. Compared with those in the control group, the ALP activity and OCN expression decreased and the Runx2 mRNA levels increased after the mTORC1 signal pathway was inhibited (P ＜ 0.001); ALP activity and OCN expression increased after the mTORC1 signal pathway was activated, while the Runx2 mRNA levels decreased (P ＜ 0.001). @*Conclusion @#The mTORC1 signaling pathway participates in the osteogenic differentiation of mouse BMMSCs under tension. The osteogenesis of BMMSCs under cyclic uniaxial tension would be enhanced if the mTORC1 signaling pathway was activated.

Exploring Mechanism of Modified Fuzi Lizhongtang in Intervening Inflammatory Response of Intestinal Mucosa in Rats with Ulcerative Colitis Based on mTOR/p-S6K1 / 中国实验方剂学杂志

Objectives:To investigate the therapeutic effect and mechanism of modified Fuzi Lizhongtang on ulcerative colitis （UC） model rats. Method:The 72 male SD rats were randomly divided into normal group，model group，sulfasalazine group（0.5 g·kg-1），modified Fuzi Lizhongtang high，medium and low-dose group （23.62，11.81，5.91 g·kg-1）. These rats were used to replicate the UC rat model by 2，4，6-trinitrobenzene sulfonic acid （TNBS）-ethanol composite modeling and treated by gavage for 2 weeks. The general condition of rats in each group was observed. After anesthesia，blood was collected from abdominal aorta and colonic tissue was taken. Semi quantitative evaluation by the colon mucosa damage index （CMDI），the pathological changes of colonic tissue were observed by the hematoxylin and eosin （HE） staining. The contents of serum interleukin-4 （IL-4），IL-6，IL-10 and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of mammalian target of rapamycin（mTOR） and phosphorylated ribosomal protein S6 kinase 1 （p-S6K1） in colonic mucosa were detected by immunohistochemistry （IHC） and Western blot. Result:Compared with normal group，the CMDI score of the model group rats was significantly increased （P<0.01）. The contents of IL-4 and IL-10 in serum were significantly decreased，the contents of IL-6 and TNF-α were significantly increased （P<0.01）. The expression levels of mTOR and p-S6K1 in colonic mucosa were up-regulated （P<0.01）. Compared with model group，the CMDI score of the modified Fuzi Lizhongtang high dose group was significantly decreased （P<0.05）. In modified Fuzi Lizhongtang high and medium dose group，the contents of IL-6 and TNF-α were significantly decreased （P<0.01） and the contents of IL-4 and IL-10 in serum were significantly increased （P<0.05，P<0.01）. In the modified Fuzi Lizhongtang high dose group，the expression level of mTOR and p-S6K1 protein was down-regulated significantly （P<0.05，P<0.01）. Conclusion:Modified Fuzi Lizhongtang high dose group can significantly reduce the congestion and edema，inflammatory cell infiltration，gland distortion，disorder of arrangement and other pathological manifestations of UC colon mucosa，and its mechanism may be related to its down-regulation of mTOR/p-S6K1 signal and the regulation of inflammatory factors secretion.

Effect of silencing mitochondrial ribosomal protein l35 gene on growth of human esophageal cancer te-1 cells / 吉林大学学报(医学版)

Objective: To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1 cells, and to clarify its mechanism Methods: Three kinds of human esophageal cancer cells, TE-1, ECA109 and KYSE150, were selected. The relative expression levels of MRPL35 mRNA in three kinds of cells by real-time quantitative PCR. The esophageal cancer TE-1 cells were divided into sh MRP 1.35 group and shCtrl group, and the cells were infcctcd with si-RNA lentivirus and si-RNA lentivirus; the esophageal cancer cell line stably silenting the MRPL35 gene was established. Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35 gene silencing. The cell growth curves in various groups were detected by CCK-8 method, and the apoptotic rates were detected by flow cytometry after Annexin \ -PE/7AAD double staining. Results: Three kinds of esophageal cancer cells expressed MRPL35 gene, and the expression levels were not statistically significant between them (P>0.05). The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35 in the TE-1 cells in shMRPI.35 group were significantly lower than those in shCtrl group ( P<∗0. 05). Compared with shCtrl group, the cell growth speed in shMRPI.35 group was decreased ( P

Effect of silencing mitochondrial ribosomal protein L35 gene on growth of human esophageal cancer TE-1 cells / 吉林大学学报(医学版)

Objective:To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1cells, and to clarify its mechanism.Methods:Three kinds of human esophageal cancer cells, TE-1, ECA109and KYSE150, were selected.The relative expression levels of MRPL35mRNA in three kinds of cells by real-time quantitative PCR.The esophageal cancer TE-1cells were divided into shMRPL35group and shCtrl group, and the cells were infected with si-RNA lentivirus and si-RNA lentivirus;the esophageal cancer cell line stably silenting the MRPL35gene was established.Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35gene silencing.The cell growth curves in various groups were detected by CCK-8method, and the apoptotic rates were detected by flow cytometry after AnnexinⅤ-PE/7AAD double staining.Results:Three kinds of esophageal cancer cells expressed MRPL35gene, and the expression levels were not statistically significant between them (P＞0.05) .The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35in the TE-1cells in shMRPL35group were significantly lower than those in shCtrl group (P＜0.05) .Compared with shCtrl group, the cell growth speed in shMRPL35group was decreased (P＜0.05) , and the apoptotic rate was significantly increased (P＜0.01) .Conclusion:Silencing MRPL35gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.

Cloning, expression, and characterization of three ribosomal protein S13 genes in Sophora flavescens / Clonagem, expressão e caracterização de três proteínas ribossômicas S13 genes em Sophora flavescens

To characterize the structure and function of ribosomal protein S13 (RPS13), we identified fulllength open reading frames (ORFs) of three RPS13 genes (RPS13-1, RPS13-2, and RPS13-3) of the Chinese medicinal plant, Sophora flavescens. The target genes were amplified by reverse transcription-olymerase chain reaction (RT-PCR), ligated into the pET22b(+) vector, and then transformed into Escherichia coli BL21 competent cells for protein expression. The physicochemical properties, protein motif, evolution, and structural organization of the three RPS13 genes were analyzed using bioinformatics tools. The full-length ORFs (453 bp) of the three RPS13 genes of S. flavescens were cloned, and each encodes a protein of 151 amino acids in length, and their expression was detected by Western blotting. Bioinformatics analysis showed that RPS13s are stable proteins that are closely related to the 40S RPS13s of Vigna radiate var. radiate. Their three-dimensional structures included three -helices at the C-terminal and four -helices at the N-terminal, and the two clusters of helices were connected by a long random coil, which may help maintain the dynamic bridging interactions between the large and small subunits of the ribosome. The full-length ORFs of three RPS13 genes of S. flavescens were successfully cloned and expressed in vitro. The study of the physicochemical properties, evolution, and secondary and three-dimensional structures of the three proteins will provide the theoretical basis for further studies on the function of RPS13s in plants.

Objetivo: Para caracterizar a estrutura e a função da proteína ribossomal S13 (RPS13), identificamos fases de leitura abertas (ORFs) completas de três genes RPS13 (RPS13-1, RPS13-2 e RPS13-3) da planta medicinal chinesa, Sophora flavescens. Métodos: Os genes alvo foram amplificados por reação em cadeia da polimerase por transcrição reversa (RT-PCR), ligados ao vetor pET22b(+), e então transformados em células competentes de Escherichia coli BL21 para expressão protéica. As propriedades físico-químicas, o motivo protéico, a evolução e a organização estrutural dos três genes RPS13 foram analisados utilizando ferramentas de bioinformática. Resultados: ORFs completos (453 pb) dos três genes RPS13 de S. flavescens foram clonados, e cada um codifica uma proteína de 151 aminoácidos de comprimento, e sua expressão foi detectada por western blotting. A análise de bioinformática mostrou que as RPS13s são proteínas estáveis que estão intimamente relacionadas com as 40S RPS13s de Vigna radiata var. radiate. Suas estruturas tridimensionais incluíam três -hélices no C-terminal e quatro -hélices no N-terminal, e os dois aglomerados de hélices eram conectados por uma longa bobina aleatória, o que pode ajudar a manter as interações de ponte dinâmicas entre o subunidades grandes e pequenas do ribossomo. Conclusões: As ORFs completas de três genes RPS13 de S. flavescens foram clonadas e expressas com sucesso in vitro. O estudo das propriedades físico-químicas, evolução e estruturas secundárias e tridimensionais das três proteínas fornecerão a base teórica para estudos adicionais sobre a função das RPS13s em plantas.

Effect of AMPK/mTOR/S6K1 pathways and the insulin-sensitizing effect for adiponectin in endometrial cancer cells / 中华妇产科杂志

Objective@#To explore effect of AMP-activated protein kinase (AMPK) /mammalian target of rapamycin (mTOR) /ribosomal protein S6 kinase-1 (S6K1) signaling pathways and the insulin-sensitizing effect by adiponectin in endometrial cancer HEC-1B cells.@*Methods@#The experiments were divided into 4 groups, adiponectin (Ad) group (HEC-1B cells treated with 20 μg/ml adiponectin for 30 minutes) , inhibitor group (HEC-1B cells treated with 10 μmol/L compound C for 30 minutes) , inhibitor+ Ad group (HEC-1B cells treated with 10 μmol/L compound C for 30 minutes following incubation with 20 μg/ml adiponectin for 30 minutes) , control group (only added the culture medium without serum DMEM) . (1) Real-time quantitative PCR and western blot analysis were used to detect the level of mRNA and protein of adiponectin receptor (AdipoR) 1 and AdipoR2. (2) Western blot analysis were used to detect phosphorylation of AMPK, mTOR, S6K1 or insulin receptor substrate 1 (IRS1) protein expression with stimulation in different concentrations of adiponectin (2.5, 5, 10, and 20 μg/ml) , or following incubation with insulin 50 nmol/L for 5 minutes; or treated with 20 μg/ml adiponectin for different times (15, 30, 45, and 60 minutes) , or following incubation with insulin 50 nmol/L for 5 minutes. (3) Cell counting kit-8 (CCK-8) assay was performed to investigate the cell proliferation, and transwell chamber assay was used to detect the cell migration in different groups.@*Results@#(1) The relative expression level of AdipoR1 mRNA and protein were higher than AdipoR2 in HEC-1B cell (8.50±0.09 to 1.00±0.00, and 0.91±0.03 to 0.69±0.03; P<0.05) . (2) The phosphorylation level of p-AMPK was significantly induced, and the phosphorylation level of p-mTOR and p-S6K1 proteins, and 20 μg/ml adiponectin at 30 minutes, AMPK protein had the highest level of activation. (3) Adiponectin induces increased tyrosine phosphorylation of IRS1 in a time-and concentration-dependent manner. (4) The proliferation inhibition ratio in Ad group (0.68±0.34) % was much more than that in inhibitor+Ad group (0.24±0.04) % (t=17.88, P<0.05) . The number of cell migration in Ad group (77±8) was much more than that in inhibitor+Ad group (132±13; t=-7.34, P<0.05) .@*Conclusions@#Adiponectin maybe inhibit proliferation and migration of endometrial cancer cells through AMPK/mTOR/S6K1 signal pathway. Adiponectin insensitizes insulin signaling may by regulating by the AMPK/S6K1/IRS1 pathway.

Ribosomal protein 16 overexpresses in prostate cancer and promotes tumor progression / 上海交通大学学报（医学版）

Objective: To investigate the effects of ribosomal protein 16 (RPS16) on tumorigenesis and development of prostate cancer. Methods: Western blotting (WB) was used to detect the differences of RPS16 levels in 25 cases of prostate cancer tissues and 33 prostate hyperplasia, and immunohistochemistry (IHC) was performed to detect RPS16 levels in 48 prostate cancer tissues and 42 benign tissues. The relationship between RPS16 level and clinical pathological parameters of the patients was analyzed. The RPS16 small interfering RNA (siRNA) was transiently transfected into DU145 and LNCaP cells by liposome method, including RPS16-siRNA1, RPS16-siRNA2 and RPS16-siRNA3. Random disturbance RPS16-siRNANC was used as negative control, cells without transfection were blank control. The efficiency of RNA interference was detected by WB 48-72 h after transfection. RPS16-siRNA with highest efficiency was chosen for subsequent cell proliferation assay, flow cytometry (FCM) and transwell assay in order to detect the effects of RPS16 on cell proliferation, cell cycle and invasion ability of DU145 and LNCaP cells. Results: WB results showed that the level of RPS16 protein in the tissues of prostate cancer was higher than that of the benign group (P=0.008). IHC results showed RPS16 protein level was significantly higher in tumor tissues than benign tissues (P=0.009). RPS16 expression was not correlated with age and metastasis, but significantly correlated with clinical stage (P=0.044) and pathological grade of the tumor (P=0.004). RPS16 siRNA can not only significantly reduce the expression of RPS16 protein in DU145 and LNCaP cells, but also inhibit the proliferation and invasion of the cancer cells, so that the cell cycle arrested in G2/M phase. Conclusion: The high expression of RPS16 protein could enhance the proliferation and invasive ability of prostate cancer cells.

Effects of Electroacupuncture on Mammalian Target of Rapamycin/70-KD Ribosomal Protein S6 Kinase Pathway in Gastrocnemius Muscle in Denervated Rats / 中国康复理论与实践

@#Objective To explore the effects and mechanism of electroacupuncture (EA) on denervation-induced atrophy in rats. Methods A total of 18 male Sprague-Dawley rats were divided into sham group (n=6), model group (n=6) and EA group (n=6). The latter two groups were clamped right sciatic nerve to establish atrophy model of skeletal muscle. On the second day after modeling, EA group accepted electroacupuncture on right Zusanli (ST36) and Huantiao (GB30) for two weeks. Their gastrocnemius muscles were obtained after intervention, and the wet weight ratio of the gastrocnemius muscles was calculated. The cross-sectional area (CSA) and diameter of muscle fibers were measured after HE staining. The protein expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), 70-KD ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6k (p-p70S6k) was tested with Western blotting. The gene expression of mTOR and p70S6K was detected with real-time quantitative polymerase chain reaction. Results Compared with the sham group, the wet weight ratio of the gastrocnemius muscle, CSA and diameter of the muscle fibers decreased in the model group and EA group (P<0.001), which were more in EA group than in the model group (P<0.01); the protein expression of mTOR, p-mTOR, p70S6K and p-p70S6K increased in the model group (P<0.01), and increased more in EA group (P<0.05); the gene expression of mTOR and p70S6K increased in the model group (P<0.05) , and increased more in EA group (P<0.05).Conclusion Electroacupuncture delays the atrophy of denervated skeletal muscles, which may relate to activation of mTOR/p70S6K signal pathway to impact synthesis of skeletal muscle proteins.

Inhibitory effect of Shiquandabu decoction on spontaneous len tumor of SV40 T antigen transgenic mice and its molecular mechanisms / 吉林大学学报(医学版)

Objective: To explore the inhibitory effect of Shiquandabu decoction on the spontaneous len tumor of the SV40 T antigen transgenic (TG) mice, and to clarify its molecular mechanism. Methods: The SV40 T antigen TG mice were randomly divided into control group (n=39) and drug treatment group (n=25). The mice in control group were fed normally, while the mice in drug treatment group were fed with Shiquandabu decoction at the 3rd week after birth, the survival time of mice was recorded. Three mice in control group and drug treatment group were randomly chosen to collect the blood from the tail vein and the amino acid levels were measured respectively 8 and 15 weeks after Shiquandabu decoction administration. Then the mice were sacrificed and the liver tissue was collected. Gene chip hybridization was used to detect the differences in the expressions of ribosomal function related genes in liver tissue of the mice in two groups and the related signal pathway was explored. Results: The survival analysis demonstrated that the survival rate of TG mice in drug treatment was higher than that in control group (P0. 05). The canonical analysis showed that thirteen genes involved in ribosomal function from 9 083 genes in liver tissue in drug treatment group had the changes compared with control group (P<0.05). Conclusion: Shiquandabu decoction can effectively prolong the lifetime of the TG mice by improving the levels of serum amino acids and promoting the liver ribosomal protein synthesis.

Danusertib induces autophagy of leukemia cells via Aurora kinase B/ribosomal protein p70S6 kinase/ribosomal protein 15 signaling pathway / 中华实用儿科临床杂志

Objective To investigate the effects of Danusertib on the changes in Aurora kinase B (Aurora B)/ribosomal protein p70S6 kinase (p70S6K)/ribosomal protein 15 (RPL15) signaling pathway and autophagy in human leukemia cells and its mechanism.Methods Myeloid leukemia cell lines THP-1 and K562 were selected as the research subjects.The experiment was divided into 2 phases.Phase 1:each cell line was treated with the concentration of Danusertib in 0.1 p mol/L,1.0 p mol/L and 5.0 μ mol/L.In control group,2 mL/L dimethyl sulfoxide (DMSO)was given.All the treated cells were cultured for 24 hours.The viability of each cell line was examined by methyhhiazoletrazolium assay and the autophagy was assessed by flow cytometry.In addition,the protein levels of p70S6K,AuroraB,phosphatidylinositol 3-kinase (PI3K),AKT(phosphorylated protein kinase B),mammalian target of rapamycin (mTOR),microtubule-associated protein (LC3),Beclin1 and RPL15 were determined by using Western blot.Part 2:Aurora B and RPL15 were down-regulated in THP-1 and K562 cells,respectively.DMSO was used to dissolve Danusertib(5.0 μ mol/L).The grouping was designed as following:DMSO group (blank control group),Danusertib treated group,empty plasmid group,small interfering RNA(siRNA) group,empty plasmid + Danusertib-treated group and siRNA + Danusertib treated group.The protein levels of Aurora B,p70S6K,RPL15,Beclinl and LC3 were detected by using Western blot.Results (1) Danusertib decreased the viability of THP-1 and K562 cells and the half maximal inhibitory concentration values were 26.9 pmol/L and 30.2 μmol/L for THP-1 and K562 cells,respectively.(2)The protein levels of p-Aurora B/Aurora B,p-p70S6K/p70S6K,RPL15,p-mTOR/mTOR and p-AKT/AKT decreased compared with control cells after being treated with 0.1 μmol/L,1.0 μ mol/L and 5.0 pmol/L of Danusertib in THP-1 and K562 cells,and the differences were statistically significant (all P ＜ 0.05).(3) In THP-1 cells,compared with the empty plasmid group,the protein levels of p70S6K and RPL15 decreased by 22.1％,61.3％ (F =18.1,P =0.001) and 55.4％,56.1％ (F =19.4,P =0.001) in siRNA group and siRNA + Danusertib-treated group after knockdown of Aurora B.In contrast,the protein levels of LC3 increased by 13.6％ and 17.1％ (F =15.4,P =0.001)compared with the empty plasmid group.In addition,the protein levels of Beclin1 and LC3 increased by 39.5％,92.3％ (F=25.2,P=0.001) and 40.2％,58.3％ (F=23.9,P=0.001) in siRNA group and siRNA + Danusertib treated group,compared with the empty plasmid group after down-regulation of RPL15.In K562 cells,compared with the empty plasmid group,the protein levels of p70S6K and RPL15 decreased by 24.2％,62.7％ (F =20.4,P=0.001) and 57.2％,60.1％ (F =23.9,P =0.001) in siRNA group and siRNA + Danusertib treated group after downregulation of Aurora B.But the protein levels of LC3 increased by 17.9％ and 56.7％ (F =20.9,P =0.001)compared with the empty plasmid group.Moreover,the protein levels of Beclin1 and LC3 were increased by 20.6％,98.4％ (F=22.4,P =0.001) and 41.5％,70.1％ (F=26.2,P =0.001) in siRNA group and siRNA + Danusertib treated group,compared with the empty plasmid group after downregulation of RPL15.Conclusion Danusertib can induce autophagy via inhibition of the PI3K/AKT/mTOR signaling pathway and can negatively regulate Aurora B/p70S6K/RPL15 axis in THP-1 and K562 cells.In addition,RPL15 may be a key target of Aurora B/p70S6K/RPL15signaling pathway in the inhibition of tumor proliferation.

Correlation of genotype, phenotype and treatment efficacy of Diamond-Blackfan anemia / 中华实用儿科临床杂志

Objective To analyze the correlation between genotype,clinical manifestations and treatment response in patients diagnosed as Diamond-Blackfan anemia (DBA) with a clear pathogenic gene mutation.Methods Retrospective investigation was performed of the genetic and clinical data of 42 patients diagnosed as DBA with a definite mutation from December 2009 to October 2017 in Beijing Children's Hospital,Capital Medical University and DBA China group.Results Among 42 patients,no one patient could successfully stop the therapy during the median follow-up time of 40 months (1-136 months).Nucleotide-level mutations or large deletions were identified in 7 ribosomal genes as RPL5,RPL11,RPL35a,RPS17,RPS19,RPS24 and RPS26.The most common gene mutation group was RPS19 (42.9％),followed by RPL11(19.0％),RPS17(11.9％),RPS26(11.9％),RPL5 (7.1％) and RPL35a(4.8％).The median onset hemoglobin level was 42.5 g/L.A total of 12 patients had physical malformation,with the most common on heart and fingers.A total of 37 patients received hormone therapy,and the total initial response rate was 89.2％ (33/37 cases).One of the patients with hormone inefficiency was treated successfully with cyclosporin A,and the other 3 patients were treated with blood transfusion.Conclusions RPS19 was the most common gene mutation in DBApatients.Most of the RPS17 mutations were copy number variation.The deletion of large fragments should be paid more attention to in the detection of DBA genetic analysis.Patients with RPL5 mutation showed more malformation than other groups.No significant difference was found in terms of age of onset,hemoglobin level of onset,incidence of malformation and effective rate of hormone treatment in each group.

Ribosomal protein 16 overexpresses in prostate cancer and promotes tumor progression / 上海交通大学学报（医学版）

Objective·To investigate the effects of ribosomal protein 16 (RPS16) on tumorigenesis and development of prostate cancer.Methods·Westem blotting (WB) was used to detect the differences of RPS16 levels in 25 cases of prostate cancer tissues and 33 prostate hyperplasia,and immunohistochemistry (IHC) was performed to detect RPS16 levels in 48 prostate cancer tissues and 42 benign tissues.The relationship between RPS16 level and clinical pathological parameters of the patients was analyzed.The RPS16 small interfering RNA (siRNA) was transiently transfected into DU145 and LNCaP cells by liposome method,including RPS16-siRNA1,RPS16-siRNA2 and RPS16-siRNA3.Random disturbance RPS16-siRNANC was used as negative control,cells without transfection were blank control.The efficiency of RNA interference was detected by WB 48-72 h after transfection.RPS16-siRNA with highest efficiency was chosen for subsequent cell proliferation assay,flow cytometry (FCM) and transwell assay in order to detect the effects of RPS 16 on cell proliferation,cell cycle and invasion ability of DU 145 and LNCaP cells.Results·WB results showed that the level of RPS 16 protein in the tissues of prostate cancer was higher than that of the benign group (P=0.008).IHC results showed RPS 16 protein level was significantly higher in tumor tissues than benign tissues (P=0.009).RPS16 expression was not correlated with age and metastasis,but significantly correlated with clinical stage (P=0.044) and pathological grade of the tumor (P=0.004).RPS 16 siRNA can not only significantly reduce the expression of RPS16 protein in DU145 and LNCaP cells,but also inhibit the proliferation and invasion of the cancer cells,so that the cell cycle arrested in G2/M phase.Conclusion·The high expression of RPS 16 protein could enhance the proliferation and invasive ability of prostate cancer cells.

Inhibitory effect of Shiquandabu decoction on spontaneous len tumor of SV40 T antigen transgenic mice and its molecular mechanisms / 吉林大学学报(医学版)

Objective:To explore the inhibitory effect of Shiquandabu decoction on the spontaneous len tumor of the SV40 T antigen transgenic (TG) mice,and to clarify its molecular mechanism.Methods:The SV40 T antigen TG mice were randomly divided into control group (n=39) and drug treatment group (n=25).The mice in control group were fed normally,while the mice in drug treatment group were fed with Shiquandabu decoction at the 3rd week after birth,the survival time of mice was recorded.Three mice in control group and drug treatment group were randomly chosen to collect the blood from the tail vein and the amino acid levels were measured respectively 8 and 15 weeks after Shiquandabu decoction administration.Then the mice were sacrificed and the liver tissue wascollected.Gene chip hybridization was used to detect the differences in the expressions of ribosomal function related genes in liver tissue of the mice in two groups and the related signal pathway was explored.Results:The survival analysis demonstrated that the survival rate of TG mice in drug treatment was higher than that in control group (P＜0.05).Compared with control group,the serum levels of alanine,valine,leucine,isoleucine,threonine,methionine,proline,tyrosine,lysine,sarcosine,citrulline,ornithine and hydroxylysine of the mice in drug treatment group 8 weeks after administration of Shiquandabu decoction were increased (P＜0.05);and the serum levels of cystathionine,taurine,methylhistidine,anserine and ethanolamine were decreased (P＜0.05).Fifteen weeks after administration,compare with control group,the serum levels of threonine and citrulline of the mice in drug teeatment group were increased (P＜0.05),but the serum levels of other amino acids had no significant difference (P＞ 0.05).The canonical analysis showed that thirteen genes involved in ribosomal function from 9 083 genes in liver tissue in drug treatment group had the changes compared with control group (P＜ 0.05).Conclusion:Shiquandabu decoction can effectively prolong the lifetime of the TG mice by improving the levels of serum amino acids and promoting the liver ribosomal protein synthesis.