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BACKGROUND:Knee osteoarthritis is a common disease in middle-aged and elderly people.It is a kind of disease that seriously affects the quality of life of patients and even has the risk of disability.Therefore,the pathogenesis and treatment of knee osteoarthritis have become the focus of research.In Chinese medicine,knee osteoarthritis is often treated as"biness,"which is closely related to"biness"caused by blood stasis and blood vessels blocking collaterals in the theory of"blood stasis"in traditional Chinese medicine.Iron overload is a kind of pathological state caused by iron metabolism disorder,which highly coincides with the pathogenic characteristics and clinical manifestations of the"blood stasis"theory of traditional Chinese medicine,and is a risk factor that promotes the development of knee osteoarthritis. OBJECTIVE:Based on the"blood stasis"theory,to summarize the effects of iron overload on cartilage metabolism and subchondral bone reconstruction,to lay a new theoretical foundation for the treatment of knee osteoarthritis with traditional Chinese medicine,and to explore the therapeutic effect of traditional Chinese medicine for promoting blood circulation after interfering with bone tissue. METHODS:CNKI,WanFang database,PubMed and Web of Science databases were searched for relevant literature.The Chinese search terms were"ferroptosis,iron,iron overload,osteoarthritis,blood stasis"and the English search terms were"ferroptosis,iron,iron overload,osteoarthritis,TCM."In the end,76 articles were included for further review. RESULTS AND CONCLUSION:First of all,we explored the potential of the"blood stasis"theory in treating knee osteoarthritis,and found that"blood stasis"is a crucial part in the progress of knee osteoarthritis,indicating that the"blood stasis"theory is the key to the treatment of knee osteoarthritis in traditional Chinese medicine.Secondly,"blood stasis"and iron overload have a high degree of similarity in pathogenic factors,clinical manifestations,and pathogenic characteristics,suggesting the possibility of"blood stasis"theory in treating iron overload.This finding reminds us that iron overload may be an important mechanistic basis for the"blood stasis"theory in the treatment of knee osteoarthritis.The extracts of blood-activating drugs can relieve iron overload and treat knee osteoarthritis,but the specific mechanism is still unclear.Therefore,we believe that the relationship between"blood stasis"theory and iron overload and related mechanisms are important research directions for knee osteoarthritis in the future.The related mechanism of"blood stasis"theory to alleviate iron overload and then treat knee osteoarthritis also provides a theoretical basis for the modernization of traditional Chinese medicine,such as the development of new drugs and innovative usage,and has certain guiding significance for clinical practice.
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Obiective Alzheimer’s disease (AD) is a degenerative disease of the central nervous system (CNS) caused by a variety of risk factors. There are various pathological changes, but apoptosis of the neurological meridian cells is one of the most important pathological bases. Hyperlipidemia is a high-risk factor for the development of AD, which can lead to increased levels of oxidized low-density lipoprotein (ox-LDL) in brain tissues. PCSK9 is a protease closely related to lipid metabolism, but studies have shown that it may be related to the development of AD. LRP1 is abundantly expressed in neuronal cells, and it is an important transporter for the clearance of Aβ. There is now a large amount of literature confirming that PCSK9 can induce the degradation of LRP1. PI3K/AKT is an important signaling pathway in vivo, which plays an important role in apoptosis, and there is now a large amount of literature confirming that LRP1 activates the PI3K/AKT pathway, which has an anti-apoptotic effect. So can PCSK9 affect the PI3K/AKT pathway through LRP1 and thus regulate neuronal apoptosis? This deserves further investigation.The aim of this study was to explore the role of PCSK9 in mediating ox-LDL pro-apoptotic neuronal cell death and its mechanism, and then further elaborate the mechanism of hyperlipidemia leading to neurodegenerative diseases such as AD. MethodsFirstly, PC12 cells were treated with different concentrations of ox-LDL (0, 25, 50, 75 and 100 mg/L) for 24 h. Oil red O staining was used to detect lipid accumulation in PC12 cells, Hoechst33258 staining and flow cytometry to detect apoptosis in PC12 cells, ELISA to detect the content of Aβ secreted by PC12, Western blot to detect expression of SREBP2, PCSK9 and LRP1. Then PC12 cells were treated with 75 mg/L ox-LDL for different times (0, 6, 12, 24, 48 h), and Western blot were performed to detect the expression of SREBP2, PCSK9 and LRP1. Finally, after transfecting 100 nmol/L PCSK9 siRNA into PC12 cells for 48 h, PC12 cells were treated with 75 mg/L ox-LDL for 24 h, Hoechst33258 staining and flow cytometry to detect apoptosis rate of PC12 cells, and Western blot to detect PCSK9, LRP1, PI3K, AKT, P-PI3K , P-AKT, NF-κB, Bcl-2, Bax, Caspase-9 and Caspase-3 expression, and ELISA detected Aβ content secreted by PC12 cells. Resultsox-LDL increased lipid accumulation and promoted apoptosis and Aβ secretion in PC12 cells, as well as increasing the expression of SREBP2 and PCSK9 and decreasing the expression of LRP1 in PC12 cells. pCsk9 siRNA could be inhibited through the PI3K/AKT pathway and the NF-κB-Bcl-2/Bax-Caspase-9/3 pathway to inhibit ox-LDL-induced apoptosis in PC12 cells while increasing Aβ secretion in PC12 cells. Conclusionox-LDL plays a bidirectional regulatory role in ox-LDL-induced apoptosis of PC12 cells by inducing an increase in PCSK9 expression and a decrease in LRP1 expression in PC12 cells, which in turn affects different signaling pathways downstream.
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OBJECTIVE@#To investigate the clinical and prognostic characteristics of primary acute myeloid leukemia (AML) with 11q23/KMT2A rearrangements.@*METHODS@#Clinical data of 90 patients with primary AML and 11q23/KMT2A rearrangements were analyzed retrospectively.@*RESULTS@#By karyotyping analysis, 80 of the 90 patients had translocations involving 11q23/KMT2A, with t(9;11)(p22;q23), t(6;11)(q27;q23), t(10;11)(p12;q23) and t(11;19)(q23;p13) being the most common ones, while 10 cases were found to have non-translocation abnormalities. The overall complete remission (CR) rate was 75.6%, and patients with t(6;11) had lower CR rate compared with non-t(6;11) patients (47.1% vs. 82.2%, P = 0.005). After a median follow-up of 24.5 months, the patients receiving allo-hematopoietic stem cell transplantation (allo-HSCT) had significantly higher 3-year overall survival (OS) (80.3% vs. 16.6%, P < 0.001) and 3-year event-free survival (EFS) (73.5% vs. 16.3%, P < 0.001) compared with non-transplant patients. Patients with t(6;11) had the lowest 3-year OS (11.8% vs. 56.0%, P < 0.001) and 3-year EFS (5.9% vs. 53.8%, P < 0.001) compared with other type of abnormalities. No significant difference was noted in the survival between patients with t(9;11) and non-t(9;11) regardless whether they had received HSCT.@*CONCLUSION@#The clinical characteristics of primary AML with 11q23/KMT2A rearrangements are heterogeneous. Patients did not receive HSCT had poorer survival, particularly with the presence of t(6;11). Allo-HSCT could significantly improve the survival of such patients.
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Humanos , Estudios Retrospectivos , Leucemia Mieloide Aguda/terapia , Translocación Genética , Reordenamiento Génico , PronósticoRESUMEN
Objective:To investigate the gene transcription level changes in the amygdala of social isolation rearing models of schizophrenia to determine the pathogenic genes and their related pathways of schizophrenia.Methods:A total of 29 3-week-old SPF C57BL/6J male mice were randomly divided into control group ( n=16) and model group ( n=13); 4 mice were raised in each transparent mouse cage in the control group, and 1 mouse was raised in each transparent mouse cage in the model group; mice in each cage could see their surrounding mice but could not touch each other. Mice in both groups were fed for 4 weeks and then subjected to open field experiment, pre-pulse inhibition experiment and new object recognition experiment within one week. After the experiment, mice were sacrificed by spinal dislocation, and the amygdala was taken for transcriptome sequencing. The topGO software was used for gene ontology (GO) functional enrichment analysis of differentially expressed genes (DEGs), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed using KEGG database. Results:(1) Animal experiment: compared with the control group, the model group had significantly increased movement distance in the open field experiment ([1 239.20±106.35] m vs. [1 845.53±143.65] m, t=3.464, P=0.002), significantly decreased activity time in the central region 5 min before experiment ([13.15±1.41] s vs. [8.47±1.19]) s, t=2.464, P=0.020). Compared with the control group, the model group had significantly lower percentage of deficient prepulse inhibition (PPI) of 78 dB ([22.28±1.53] % vs. [14.59±2.75] %, t=2.629, P=0.013), and deficient PPI of 88 dB ([32.83±3.39] % vs. [18.44±3.07] %, t=3.081, P=0.005). Compared with the control group, the model group had significantly decreased ratio of time exploring new objects/time exploring former objects ([80.5±2.2]% vs. [71.0±3.6]%, t=2.356, P=0.026). (2) Bioinformatics analysis: a total of 96 DEGs were found, of which 42 were with up-regulated expressions ( Th, Crlf1, etc.), and 54 were with down-regulated expressions ( Prkcd, etc.). Th and Crlf1 were positively correlated ( r=0.940, P=0.018). GO enrichment results suggested that DEGs were enriched in projection function of plasma membrane boundary cells, neuronal differentiation, and cell apoptosis. KEGG enrichment results suggested that DEGs were enriched in WNT signaling pathway, apoptosis pathway and tyrosine metabolism pathway. Protein network interaction analysis suggested that Wnt6, Tcf712, Pitx2, Tcf7 and Cd4 were key proteins. Conclusion:DEGs such as Th, Prkcd, Lrrc74b, Fadd, Wnt6, Ror2, Notum, and Tcf7l2, and their related signaling pathways may be related to schizophrenia in the amygdala of social isolation rearing mice.
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AIM:To explore the correlation and consistency of three tear assessment methods in the diagnosis of dry eye, which include the Schirmer Ⅰ test(SⅠt), tear meniscus height(TMH)measurement and a new generation of tear detection technology-Strip meniscometry tube(SMTube).METHODS: A diagnostic test study. A total of 183 dry eye outpatients(183 right eyes)in the Dry Eye Center of Henan Eye Hospital were enrolled from May to June 2021. The SⅠt, TMH and SMTube were performed on all patients, and the correlation and consistency of the measurement results were analyzed.RESULTS:The ocular surface disease index(OSDI)of all included patients was 43.75(31.25, 58.33), noninvasive breakup time(NIBUT)was 7.26(4.97, 9.37)s, and the results of SⅠt, TMH and SMTube were 6(2, 12)mm/5min, 0.18(0.14, 0.22)mm and 5(3,8)mm/5s, respectively; The results of correlation analysis: TMH was positively correlated with SMTube(rs=0.751, P<0.001), however, SⅠt had no correlation with TMH(rs=0.139, P=0.060)and SMTube(rs=0.019, P=0.799). The results of consistency analysis: TMH showed good consistency with SMTube(Kappa=0.794, P=0.044), however, SⅠt showed poor consistency with TMH(Kappa=0.271, P=0.074)and SMTube(Kappa=0.193, P=0.070)respectively.CONCLUSION:SMTube is a new, simple and convenient tool for evaluating tear volume. It has a good correlation and consistency with TMH measurement results. Therefore, it can replace TMH measurement, but cannot replace SⅠt in clinical application.
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Ulcerative colitis (UC) is a chronic non-specific inflammatory disease characterized by the damage of the epithelial barrier of the colon and the destruction of immune homeostasis. It has a long course, no recovery and high recurrence rate, and is recognized as a difficult digestive disease. MicroRNA (miRNA) has been confirmed to be specifically or differentially expressed in both UC patients and UC animal models, so miRNA can be used as markers for UC diagnosis or reference for treatment evaluation. TCM therapy has a definite therapeutic effect, a wide range of effects, and minimal side effects in the treatment of UC, so this article takes miRNA as the starting point and systematically elaborates on the mechanism of TCM regulating UC related signaling pathways by regulating the expression of miRNA. The results show that chlorogenic acid, Anchang decoction, and Fuyang huoxue jiedu formula can regulate the expressions of miR-155, miR-146a and miR-31-5p, etc., thereby inhibiting signal transducer and activator of transcription (STAT) signal pathway transduction to improve UC. Limonin, ginsenoside Rh2, artesunate, etc. can inhibit nuclear factor-κB (NF-κB) signaling pathway conduction to improve UC by regulating the expressions of miR-214, miR- 155 and miR-19a, etc. Nitidine chloride, berberine, resveratrol, etc. can regulate the expressions of miR-31, miR-146a, miR- 146b, etc., thereby inhibiting the Toll-like receptor 4 (TLR4) signaling pathway to improve UC. Mango polyphenolics, Compound qinbai granules, and Astragalus membranaceus polysaccharides can regulate the expressions of miR-126 and miR-193a-3p, thereby inhibiting the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway to improve UC.
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Aim To study the inhibitory effect of ginsenoside Rgl on neuronal ferroptosis after ischemic stroke and its mechanism. Methods A model of oxygen glucose deprivation/reoxygenation (OGD/R) was established in HT22 cells, and the effect of Rgl on the viability of HT22 cells after OGD/R injury was detected by CCK-8. The effect of Rgl on ferroptosis in HT22 cells after OGD/R injury was detected by the test of ferroptosis markers GSH/GSSG, SOD, MDA, and Fe
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ObjectiveTo investigate the effect of Biejiajian Wan (BJJW) on transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of HepG2 cells, and explore its mechanism against EMT of hepatocellular carcinoma cells. MethodHepG2 cells were randomly divided into a blank group, a TGF-β1 model group (10 μg·L-1 TGF-β1), a low-dose BJJW group (10 μg·L-1 TGF-β1+0.55 g·kg-1 BJJW), a medium-dose BJJW group (10 μg·L-1 TGF-β1+1.1 g·kg-1 BJJW), a high-dose BJJW group (10 μg·L-1 TGF-β1+2.2 g·kg-1 BJJW), and a sorafenib group (10 μg·L-1 TGF-β1+0.03 g·kg-1 sorafenib). The EMT model was induced by 10 μg·L-1 TGF-β1 in HepG2 cells. After treatment with corresponding medicated serum, cell counting kit -8 (CCK-8) assay was used to detect cell proliferation. Cell migration ability was detected by the Transwell assay and wound healing assay. The protein expression related to EMT and nuclear factor-kappa B (NF-κB) signaling pathway was detected by cell immunofluorescence assay and Western blot. ResultCompared with the blank group 4 days later, the TGF-β1 model group showed fusiform and loose cells with widened gap and antennae reaching out, decreased protein expression of E-cadherin (P<0.05), and increased protein expression of N-cadherin and vimentin (P<0.05), which indicated that the EMT model was properly induced in HepG2 cells by TGF-β1 stimulation for 4 days. After 48 hours of treatment with the corresponding medicated serum, each medication group showed inhibited proliferation of HepG2 cells that had undergone EMT, especially the low- and high-dose BJJW groups (P<0.01), and the medium-dose BJJW group showed increased E-cadherin protein expression (P<0.05) and decreased p-p65, N-cadherin, and vimentin protein expression (P<0.05), as compared with the TGF-β1 model group. As revealed by the transwell assay and wound healing assay, TGF-β1 enhanced the migration ability of HepG2 cells (P<0.05, P<0.01) compared with the results in the blank group, compared with the TGF-β1 model group, the medication groups showed inhibited migration ability of HepG2 cells (P<0.05, P<0.01). Compared with the blank group, the TGF-β1 model group promoted the expression of p65 and Snail into the nucleus. Compared with the TGF-β1 model group, the medication groups inhibited the expression of p65 and Snail into the nucleus. ConclusionBJJW may inhibit the EMT, proliferation, and migration of HepG2 cells induced by TGF-β1 by suppressing the NF-κB signaling pathway to exert an anti-hepatocellular carcinoma effect.
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ObjectiveTo investigate the compatibility rule of traditional Chinese patent medicines (TCPMs) against liver diseases through network analysis. MethodWith “liver” as the search term, TCPMs against liver diseases were retrieved from volume Ⅰ of Chinese Pharmacopoeia (2020 edition), and the basic information of them was collected. TCPMs with same Chinese medicinal materials (CMMs), usage, and indications, but different dosage forms, were unified as one formula. Mutual information entropy (MIE) of CMM couples was calculated to quantify the relationship between them, and the top 25% CMM pairs in MIE were used to construct the compatibility network, with CMM as node and the relationship between CMM pairs as the edge. Key CMM and frequently used CMM combinations were identified based on node centrality and cluster analysis, respectively. The indications of TCPMs related to the CMMs in clusters were recorded. Cytoscape 3.6.1 was employed for visualization and topology analysis of the compatibility network. ResultA total of 179 TCPMs, involving 428 CMMs, were retrieved. Angelicae Sinensis Radix, Paeoniae Radix Alba, and Glycyrrhizae Radix et Rhizoma were identified as key CMMs with high frequency, and Cuscutae Semen-Lycii Fructus, Citri Reticulatae Pericarpium-Cyperi Rhizoma, and Ecliptae Herba-Ligustri Lucidi Fructus combinations had high MIE. Furthermore, the CMMs were clustered into ten groups corresponding to different diseases which, however, all belonged to digestive diseases. ConclusionThis study unveils potential CMM pairs and common CMM combinations against liver diseases, which can serve as a reference for revealing compatibility rules of CMMs and research and development of Chinese medicine.
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ObjectiveTo explore the effect of Jinshui Xiangsheng prescription on the five-year clinical survival outcome of patients with advanced prostate cancer. MethodFrom May 1, 2014 to May 1, 2016, patients with advanced prostate cancer from Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine and the Urology Department of the Second Affiliated Hospital of Nanjing University of Chinese Medicine were collected and treated with Jinshui Xiangsheng prescription (155 cases in the observation group). According to age and Gleason score, the patients without Jinshui Xiangsheng prescription were matched in a ratio of 1∶1 (155 cases in the control group). The androgen resistance rate, survival rate, median survival time, and median progress free survival time in 1, 3, 5 years were observed. The prognostic factors of advanced prostate cancer were analyzed and screened out by Chi-square test, t test, Kaplan-Meier and Cox survival analysis. ResultThe androgen resistance rates in the observation group in 1, 3, 5 years were 9.0% (14/155), 72.3% (112/155), and 92.9% (144/155), respectively, and those in the control group were 20.6% (32/155), 87.7% (136/155), and 97.4% (151/155). The 1-year (χ2=8.271,P<0.01)and 3-year (χ2=11.613,P<0.01) androgen resistance rates in the observation group were significantly lower than those in the control group. The median survival time and median progress free survival time in the observation group were (26.35±9.01) months and (11.02±4.40) months, respectively, and in the control group were (22.31±9.21) months and (9.87±5.12) months, respectively. The median survival time and median progress free survival time in the observation group were significantly longer than those in the control group (P<0.05, P<0.01). The cumulative survival rates in 1, 3, 5 years in the observation group were 96.1% (149/155), 80.6% (125/155), and 39.4% (61/155), respectively, and those in the control group were 94.2% (146/155), 60.0% (93/155), and 22.6% (35/155), respectively. The 3-year (χ2=15.828,P<0.01) and 5-year (χ2=10.201,P<0.01) cumulative survival rates in the observation group were significantly higher than those in the control group. The monofactor analysis showed that the prognostic factors involved in Gleason score, initial prostate specific antigen (PSA), tumor location, tumor stage, castration regimen, radiotherapy, chemotherapy, complete androgen blockade (CAB), and Jinshui Xiangsheng prescription (P<0.05, P<0.01). The multivariate analysis showed that initial PSA, tumor location, and tumor stage were the risk factors affecting the survival time of patients with advanced prostate cancer, whereas Jinshui Xiangsheng prescription, castration regimen, chemotherapy, radiotherapy, and CAB were the independent protective factors affecting the prognosis of advanced prostate cancer. ConclusionJinshui Xiangsheng prescription has a protective effect on the survival of patients with advanced prostate cancer, which reduces the androgen resistance rate and death risk of advanced prostate cancer, thus benefiting the survival of patients. Therefore, it deserves further promotion.
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Aim To construct Flp-In CHO cell line(CYP2A13-CHO)stably expressing cytochrome P450 family 2 subfamily A member 13(CYP2A13)and Flp-In CHO cell line(CYP2A13-POR-CHO)stably co-expressing CYP2A13 and cytochrome P450 oxidoreductase(POR), from which a cell line with better metabolic activity is selected.Method In our previous study, we had constructed a Flp-In CHO cell line(POR-Flp-In CHO)stably expressing POR using lentiviral vector.The recombinant plasmids of pcDNA5/FRT-CYP2A13 were constructed and transfected into Flp-In CHO cells and POR-Flp-In CHO cells through LipofectamineTM 2000.The expression and activity of CYP2A13 were detected by real-time quantitative PCR(qRT-PCR), Western blot and Aflatoxin B1(AFB1)/4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)cytotoxicity assay and the metabolic activity was compared between CYP2A13-CHO and CYP2A13-POR-CHO.Results Compared with non-transfected cells, the mRNA and protein expression of CYP2A13 in CYP2A13-CHO and CYP2A13-POR-CHO cells both increased significantly.Besides, compared with CYP2A13-POR-CHO, CYP2A13-CHO cells were more sensitive to AFB1 and NNK.Conclusions The Flp-In CHO cell line stably expressing CYP2A13 and with better metabolic activity has been established successfully, which provides a tool for screening of pre-carcinogens that can be metabolically activated by CYP2A13.
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Objective To investigate the effects and molecular mechanism of circular RNA (circ) _0003028 on proliferation, migration and invasion of human liver cancer cells. Methods Liver cancer cell line Huh7 were divided into small interfering RNA(si)-NC group, si-circ_0003028 group, microRNA(miR) -NC group, miR-498 mimics group, si-circ _0003028+anti-miR-NC group and si-circ_0003028+anti-miR-498 group. Real-time PCR was used to detect the expression levels of circ _ 0003028 and miR-498 in liver cancer tissues and cells of each groups. MTT was used to detect cell proliferation. Transwell was used to detect cell migration and invasion. Western blotting was used to detect protein expression. Dual luciferase reporter experiment was used to detect the target regulation relationship between circ_0003028 and miR-498. Results The expression level of circ_0003028(0.98±0.02 vs 1. 36±0. 01) increased and the expression level of miR-498(0. 98±0. 02 vs 0. 63±0. 02) decreased in liver cancer tissues (P<0. 05). After inhibiting the expression of circ_0003028 or overexpression of miR-498- the expression levels of Ki-67(0. 85±0. 02 vs 0. 41±0. 02 or 0. 95±0. 11 vs 0. 37±0. 02)- matrix metalloprotein(MMP)-2(0. 71±0. 02 vs 0. 43±0. 03 or 0. 83±0. 02 vs 0. 41±0. 03)- and MMP-9 (0. 74±0. 02 vs 0. 37±0. 02 or 0. 78±0. 02 vs 0. 39±0. 02) proteins in Huh7 cells decreased- and cell viability(1. 53± 0. 03 vs 1. 05±0. 02 or 1. 68±0. 02 vs 1. 11±0. 02) decreased- the number of migration(111. 40±2. 12 vs 77. 22±2. 38 or 108. 90±2. 30 vs 78. 44 ± 1. 46) and invasion ( 87. 89 ± 2. 18 vs 49. 78 ± 1. 98 or 80. 22 ± 1. 79 vs 38. 22 ± 1. 52) cells decreased- and the protein expression levels of suppressor with morphogenesis effect on genitalia-1( SMG-1) (0. 76±0. 02 vs 1. 39±0. 02 or 0. 79±0. 02 vs 1. 39±0. 02)- p53(0. 77±0. 02 vs 1. 24±0. 03 or 0. 82±0. 03 vs 1. 45±0. 03)- and p53- ser15(0. 78±0. 03 vs 1. 50±0. 02 or 0. 82±0. 02 vs 1. 49±0. 04) increased (P<0. 05). circ_0003028 targeted regulation of miR-498- and silencing miR-498 reversed the effects of inhibiting the expression of circ_0003028 on the proliferation- migration and invasion of Huh7 cells. Conclusion Inhibiting the expression of circ_0003028 can inhibit the proliferation- migration and invasion of liver cancer cells by targeting miR-498 to affect the SMG-1/ p53 signaling pathway.
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Gloeostereum incarnatum has edible and medicinal value and was first cultivated and domesticated in China. We sequenced the G. incarnatum monokaryotic strain GiC-126 on an Illumina HiSeq X Ten system and obtained a 34.52-Mb genome assembly sequence that encoded 16,895 predicted genes. We combined the GiC-126 genome with the published genome of G. incarnatum strain CCMJ2665 to construct a genetic linkage map (GiC-126 genome) that had 10 linkage groups (LGs), and the 15 assembly sequences of CCMJ2665 were integrated into 8 LGs. We identified 1912 simple sequence repeat (SSR) loci and detected 700 genes containing 768 SSRs in the genome; 65 and 100 of them were annotated with gene ontology (GO) terms and KEGG pathways, respectively. Carbohydrate-active enzymes (CAZymes) were identified in 20 fungal genomes and annotated; among them, 144 CAZymes were annotated in the GiC-126 genome. The A mating-type locus (MAT-A) of G. incarnatum was located on scaffold885 at 38.9 cM of LG1 and was flanked by two homeodomain (HD1) genes, mip and beta-fg. Fourteen segregation distortion markers were detected in the genetic linkage map, all of which were skewed toward the parent GiC-126. They formed three segregation distortion regions (SDR1–SDR3), and 22 predictive genes were found in scaffold1920 where three segregation distortion markers were located in SDR1. In this study, we corrected and updated the genomic information of G. incarnatum. Our results will provide a theoretical basis for fine gene mapping, functional gene cloning, and genetic breeding the follow-up of G. incarnatum.
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Objective:To study the effect of Biejiajian Wan on the epithelial-mesenchymal transition (EMT) of rat hepatic oval cells induced by transforming growth factor- β1(TGF-β1), in order to explore its mechanism in reversing EMT. Method:WB-F344 cells were divided into five groups: blank group, TGF-β1 model group (10 μg·L-1TGF-β1), low-dose group (10 μg·L-1TGF-β1+0.55 g·kg-1Biejiajian Wan), medium-dose group (10 μg·L-1TGF-β1+1.1 g·kg-1Biejiajian Wan), high-dose group (10 μg·L-1TGF-β1+2.2 g·kg-1Biejiajian Wan). Except blank group, TGF-β1 was used to induce WB-F344 cells in all of the remaining groups to construct an EMT model. After the cells were treated with low, medium and high doses of Biejiajian Wan serum, the changes of migration ability of WB-F344 cells were detected by cell scratching test. The expressions of E-cadherin, N-cadherin and Vimentin were detected by Western blot. Real-time PCR was used to detect the changes in the expression of β-catenin mRNA. The expression of β-catenin was detected by cell immunofluorescence assay. Result:Compared with normal WB-F344 cells, the intercellular space of WB-F344 cells became loose from tight, and the morphology changed from cobblestone to fibroblast after TGF-β1 induced WB-F344 cells for 4 days, and the expression of E-cadherin protein decreased, while the expression of N-cadherin protein increased (P<0.01), indicating that the EMT model of WB-F344 cells was successfully built. Compared with the blank group, the migration ability of WB-F344 cells in TGF-β1 model group was enhanced (P<0.01), compared with TGF-β1 model group, Biejiajian Wan could significantly inhibit the migration ability of WB-F344 cells; specifically, the low-dose group had no statistical significance, and the medium and high-dose groups had statistical significance (P<0.05). Western blot results showed that compared with the blank group, the expression of E-cadherin decreased, whereas those of N-cadherin and Vimentin increased in the TGF-β1 model group (P<0.01), compared with TGF-β1 model group, E-cadherin protein expression was increased in the low, medium and high-dose groups, while the expressions of N-cadherin and Vimentin was decreased; specifically, the low-dose groups had no statistical significance, and the medium and high dose groups had statistical significance (P<0.05,P<0.01). Real-time PCR results showed that compared with the blank group, the mRNA expression of β-catenin in the TGF-β1 model group was increased (P<0.05), whereas compared with TGF-β1 model group, the mRNA expression of β-catenin in the low, medium and high-dose groups of Biejiajian Wan was reduced (P<0.01). The results of cellular immunofluorescence showed that compared with the blank group, the fluorescence expression of β-catenin in the cell nucleus was enhanced in the TGF-β1 model group; and compared with the TGF-β1 model group, the expression of β -catenin in the cell nucleus of the low, medium and high-dose groups of Biejiajian Wan decreased, and the inhibitory effect of Biejiajian Wan on β-catenin in the cell nucleus was positively correlated with its concentration. Conclusion:Biejiajian Wan may reverse the EMT process that TGF-β1 induced WB-F344 cells, and inhibit the migration of WB-F344 cells by inhibiting Wnt/β-catenin signaling pathway.
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Objective:To investigate the risk factors related to extrathyroidal extension (ETE) of differentiated thyroid carcinoma (DTC) and the specific effects on the prognosis.Methods:The clinical data of 592 patients with newly diagnosed DTC admitted to our hospital from Jun. 2015 to Jun. 2016 were retrospectively analyzed. The data including the maximum tumor diameter and lymph node metastasis were collected, and the survival data were followed up. Chi-square test and Logistic regression were used to analyze the risk factors associated with ETE. Kaplan-Meier method and Cox proportional risk model were used to analyze the effect of ETE on disease-free survival.Results:There were 100 ETE of 592 DTC patients (16.9%) . Univariate analysis showed that the risk factors for ETE were the largest tumor diameter ≥2 cm, multiple lesions, and lymph node metastasis in the lateral cervical region ( P<0.01) . The follow-up time was 29 to 64 months, and the median follow-up time was 60 months. The 3-year and 5-year DFS rates of patients with DTC and ETE were 86.9% and 83.2% respectively, which were significantly lower than those of patients without ETE ( P<0.001) . Univariate analysis showed that ETE ( P<0.001) was a risk factor for postoperative recurrence in patients with DTC. Cox multivariate regression showed that ETE (HR: 10.564, 95% CI 3.712-30.063, P<0.001) was an independent risk factor for postoperative recurrence in patients with DTC. Conclusions:DTC accompanied by ETE is associated with risk factors such as large tumor diameter and multiple lesions, which has a lower 5-year DFS. Patients with the above characteristics should consider the possibility of ETE before surgery, and focus on long-term follow-up after surgery.
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OBJECTIVE@#To observe the effectiveness and safety of electrothermal acupuncture therapy for patients of moderate to severe cancer pain with @*METHODS@#A total of 60 patients of moderate to severe cancer pain with @*RESULTS@#The variation of NRS scores in the observation group were larger than the control group 3, 5 days into treatment (@*CONCLUSION@#On the basis of the conventional western medication for analgesia, electrothermal acupuncture could relieve pain, reduce the dose of opioid painkillers and improve the quality of life in patients of moderate to severe cancer pain with
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Humanos , Puntos de Acupuntura , Terapia por Acupuntura , Dolor en Cáncer/terapia , Neoplasias/terapia , Oxicodona , Calidad de Vida , Resultado del TratamientoRESUMEN
Gloeostereum incarnatum has edible and medicinal value and was first cultivated and domesticated in China. We sequenced the G. incarnatum monokaryotic strain GiC-126 on an Illumina HiSeq X Ten system and obtained a 34.52-Mb genome assembly sequence that encoded 16,895 predicted genes. We combined the GiC-126 genome with the published genome of G. incarnatum strain CCMJ2665 to construct a genetic linkage map (GiC-126 genome) that had 10 linkage groups (LGs), and the 15 assembly sequences of CCMJ2665 were integrated into 8 LGs. We identified 1912 simple sequence repeat (SSR) loci and detected 700 genes containing 768 SSRs in the genome; 65 and 100 of them were annotated with gene ontology (GO) terms and KEGG pathways, respectively. Carbohydrate-active enzymes (CAZymes) were identified in 20 fungal genomes and annotated; among them, 144 CAZymes were annotated in the GiC-126 genome. The A mating-type locus (MAT-A) of G. incarnatum was located on scaffold885 at 38.9 cM of LG1 and was flanked by two homeodomain (HD1) genes, mip and beta-fg. Fourteen segregation distortion markers were detected in the genetic linkage map, all of which were skewed toward the parent GiC-126. They formed three segregation distortion regions (SDR1–SDR3), and 22 predictive genes were found in scaffold1920 where three segregation distortion markers were located in SDR1. In this study, we corrected and updated the genomic information of G. incarnatum. Our results will provide a theoretical basis for fine gene mapping, functional gene cloning, and genetic breeding the follow-up of G. incarnatum.
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The existence of cancer stem cells is regarded as the major cause for therapeutic resistance and relapse of a variety of cancer types including hepatocellular carcinoma (HCC). However, the tracing of such a subpopulation in vivo has been challenging. We have previously demonstrated that the isoform 5 of the voltage-gated calcium channel α2δ1 subunit, which can be recognized specifically by a monoclonal antibody 1B50-1, is a bona fide surface marker for HCC stem cells. Here we developed a strategy for optical imaging of α2δ1-positive cells by using a fusion protein containing the single chain variable fragment (scFv) of Mab1B50-1 and the luciferase NanoLuc which was tagged with Flag in the C-terminal. The scFv of Mab1B50-1 was fused to the N-terminal of NanoLucFlag using overlap PCR, and the recombinant fragment, which was named as 1B50-1scFv-NanoLucFlag, was subsequently cloned into a eukaryotic expression vector. The resulting construct was transfected into FreeStyle 293F cells in suspension using PEI reagent. The expression of the fusion protein was identified as a protein with molecular weight about 50 kDa by Western blotting. After purification by ANTI-FLAG® M2 affinity chromatography, 1B50-1scFv-NanoLucFlag was demonstrated to bind to α2δ1 positive cells specifically with a Kd value of (18.62±1.84) nmol/L. Furthermore, a strong luciferase activity of 1B50-1scFv-NanoLucFlag was detected in α2δ1 positive cells following incubation with the fusion protein, indicating that the presence of α2δ1 could be quantified using this fusion protein. Hence, 1B50-1scFv-NanoLucFlag provides a potential tool for optical imaging of α2δ1 positive cancer stem cells both in vitro and in vivo.
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Humanos , Carcinoma Hepatocelular , Neoplasias Hepáticas , Células Madre Neoplásicas , Proteínas Recombinantes/genética , Anticuerpos de Cadena Única/genéticaRESUMEN
Objective:To observe the expression levels of glial fibrillary acidic protein (GFAP), β-tubulin Ⅲ and synaptophsin, and explore the role of tripartite synapse in the mechanism of central nervous system (CNS) injury and the neuroprotective effect of chondroitin sulfate (CS).Methods:One month old clean grade, 48 female Sparague-Dawley rats and 48 male Sparague-Dawley rats, were randomly divided into 8 groups according to body weight (90 - 120 g) by random number table method, with 12 rats in each group, half male and half female. These rats were fed with water containing different concentrations of sodium fluoride (NaF) [ < 0.5 mg/L (control, CN), 10.0 mg/L (low dose fluoride, LF) and 50.0 mg/L (high dose fluoride, HF)]. Some rats were fed directly for 185 days (CN, LF and HF groups). In addition, rats of CN + normal saline (NS), LF + NS, HF + NS groups and LF + CS, HF + CS groups, were intraperitoneally injected with NS or 0.66 mg/kg CS for 5 consecutive days after 180 days of feeding. After the experiment, the pathological changes of hippocampal CA4 of brain tissue in each group were observed by hematoxylin eosin staining under light microscope, and the expression and distribution of GFAP, β-tubulin Ⅲ and synaptophsin in hippocampal CA4 of rats were detected by immunohistochemistry, the expression of GFAP, β-tubulin Ⅲ and synaptophsin at protein level in hippocampus of rats were detected by Western blotting.Results:Under light microscope, eosinophilic change, loss and irregular arrangement of neuron in the hippocampal CA4 were observed in LF, HF, LF + NS and HF + NS groups. The morphology of LF + CS and HF + CS groups was not significantly changed compared with CN group, but was significant changed compared with LF, HF, LF + NS and HF + NS groups. Immunohistochemical results showed that the rates of positive area of GFAP, β-tubulin Ⅲ and synaptophsin in female and male rats in LF and HF groups were significantly decreased than those in CN group ( P < 0.05); the positive area rates of female and male rats in LF + CS and HF + CS groups were higher than those in LF and HF groups, respectively ( P < 0.05). Western blotting results showed that the proten expression levels of GFAP, β-tubulin Ⅲ and synaptophsin of female and male rats in LF and HF groups (LF group: 0.90 ± 0.09, 0.82 ± 0.08, 1.43 ± 0.14, 0.92 ± 0.02, 1.21 ± 0.15, 0.87 ± 0.02, HF group: 0.58 ± 0.14, 0.73 ± 0.03, 0.63 ± 0.06, 0.67 ± 0.03, 0.87 ± 0.04, 0.70 ± 0.05) were lower than those in CN group (1.24 ± 0.08, 1.09 ± 0.10, 2.64 ± 0.30, 1.54 ± 0.09, 1.72 ± 0.10, 1.13 ± 0.06, P < 0.05). Conclusions:The tripartite synapse and extracellular matrix may take part in pathogenesis of the damages of CNS results from chronic fluorosis; CS may reduce the injury to a certain extent.
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Objective: To compare the recovery and quality of life of patients with oral and oropharyngeal tumors treated with three kinds of free soft tissue flaps. Methods: The clinical data of 103 patients, including 66 males and 37 females, aged 26-74 years, who underwent primary repair of defects after resection of oral and oropharyngeal tumors in Sichuan Tumor Hospital from July 2014 to August 2020 were analyzed. Anterolateral thigh flap (ALTF) was used in 43 patients, radial forearm free flap (RFFF) in 45 patients, and lateral arm free flap (LAFF) in 15 patients. Postoperative qualities of life of patients were evaluated by the university of Washington quality of life questionnaire and oral health impact scale (HIP-14 Chinese edition). SPSS 23.0 software was used for statistical analysis. Results: The T staging of RFFF or LAFF group was significantly lower than that of ALTF group (P<0.05). There was no significant difference in mean flap areas between ALTF group ((55.87±27.38) cm2) and LAFF group ((49.93±19.44) cm2), while RFFF group had smaller mean flap area ((33.18±6.05) cm2) than ALTF group (t=5.311, P<0.001) and LAFF group (t=3.284, P=0.005). In terms of oral functions including swallowing, mastication, taste and spitmouth, there were no significant differences between LAFF group and RFFF group (P>0.05), but both groups had better oral functions than ALTF group (P<0.05). There was no significant difference in appearance scores between LAFF group (75(75, 75)) and ALTF group (75(75,75) vs.75(75,75),Z=-1.532, P=0.126), and both groups had higher scores than RFFF group (50(50, 75),Z values were -3.447 and -3.005 respectively, P<0.05). RFFF group had higher speech score (100(67, 100)) than LAFF group (67(50, 76),Z=-2.480, P<0.05) and ALTF group (67(33, 67),Z=-5.414, P<0.05). ALTF group had lower mean score of quality of life than RFFF group [72(56,77) vs.79(69, 89),Z=-3.070, P<0.05), but there was no statistical difference in the mean scores of qualities of life between ALTF group and LAFF group (Z=1.754, P=0.079). According to the evaluation of oral health impact scale (HIP-14 Chinese version) 1 year after surgery, individual item scores and the average score of all items in ALTF group were lower than those in RFFF and LAFF groups (P<0.05), with no significant difference between RFFF group and LAFF group (P>0.05). Conclusions: RFFF has unique advantages for small tissue defects, while ALTF is suitable for large tissue defects, such as buccal penetrating defect, whole tongue and near whole tongue defect, and LAFF is a compromise choice between ALTF and RFFF. ALTF is inferior to RFFF and LAFF in oral functional reconstruction, including swallowing, chewing, taste and spittle. ALTF and LAFF are superior to RFFF in postoperative appearance.