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OBJECTIVES@#Non-small cell lung cancer (NSCLC) accounts for 85% of all lung cancer, with highmorbidity and mortality rate. Nove drug development for NSCLC is urgently needed.This study aims to investigate the activity of lathyrol derivatives and the mechanism for its inhibitory effect on the growth of NSCLC cells.@*METHODS@#Three lathyrol derivatives were synthesized from lathyrol and their structures were verified by nuclear magnetic resonance. MTT assay was used to detect the effects of the lathyrol derivatives on the proliferation activity of NSCLC cells (A549 and H1299 cells), and the compound with the best activity was selected for subsequent experiments. Colony forming assay, wound-healing assay, and transwell assay were applied to detect in vitro cell proliferation, migration and invasion ability in A549 and H1299 cells, respectively. Quantitative real-time RT-PCR and Western blotting were performed to detect mRNA and protein levels of E-cadherin, N-cadherin, β-catenin, and MMP2 in A549 cells, respectively.@*RESULTS@#Three lathyrol derivatives inhibited the growth of A549 and H1299 cells in a dose-dependent manner, and they showed a weak inhibitory effect on normal cells Beas-2B and 16HBE, indicating that they possessed certain selective toxic effects. Therefore, C-5 benzoylated lathyrol with the best activity was selected as the ideal drug for the subsequent experiments. Compared with the control group, the number and size of cell clusters in the treatment group of A549 and H1299 cells were significantly decreased, the relative mobility were significantly decreased, and the number of invaded cells were significantly decreased (all P<0.05), indicating that the in vitro cell proliferation, migration and invasion ability were decreased. The mRNA levels of integrin α2, integrin β1, MMP2, MMP9, β-catenin, and N-cadherin were decreased, while the expression of E-cadherin was increased (all P<0.05). The protein levels of N-cadherin, β-catenin, MMP2, and integrin αV were decreased, while the expression of E-cadherin was increased (all P<0.05).@*CONCLUSIONS@#The lathyrol derivatives synthesized in this study possess good inhibitory activity against NSCLC. Among them, C-5 benzoylated lathyrol significantly inhibits the proliferation, migration, and invasion ability of NSCLC cells in vitro through regulating the process of epithelial-mesenchymal transition.
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Humans , Cadherins/genetics , Carcinoma, Non-Small-Cell Lung/drug therapy , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition , Lung Neoplasms/drug therapy , Matrix Metalloproteinase 2/genetics , RNA, Messenger , beta Catenin/geneticsABSTRACT
Objective@#Abnormalities of static brain activity have been reported in schizophrenia, but it remains to be clarified the temporal variability of intrinsic brain activities in schizophrenia and how atypical antipsychotics affect it. @*Methods@#We employed a resting-state functional magnetic resonance imaging (rs-fMRI) and a sliding-window analysis of dynamic amplitude of low-frequency fluctuation (dALFF) to evaluate the dynamic brain activities in schizophrenia (SZ) patients before and after 8-week antipsychotic treatment. Twenty-six schizophrenia individuals and 26 matched healthy controls (HC) were included in this study. @*Results@#Compared with HC, SZ showed stronger dALFF in the right inferior temporal gyrus (ITG.R) at baseline. After medication, the SZ group exhibited reduced dALFF in the right middle occipital gyrus (MOG.R) and increased dALFF in the left superior frontal gyrus (SFG.L), right middle frontal gyrus (MFG.R), and right inferior parietal lobule (IPL.R). Dynamic ALFF in IPL.R was found to significant negative correlate with the Scale for the Assessment of Negative Symptoms (SANS) scores at baseline. @*Conclusion@#Our results showed dynamic intrinsic brain activities altered in schizophrenia after short term antipsychotic treatment. The findings of this study support and expand the application of dALFF method in the study of the pathological mechanism in psychosis in the future.
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BACKGROUND: Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. METHODS: We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose–derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10 days, cells adhesion and differentiation was evaluated. RESULTS: Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. CONCLUSION: In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.
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Animals , Humans , Rats , Blotting, Western , Fluorescent Antibody Technique , In Vitro Techniques , Kidney , Mesoderm , Nephrons , Regeneration , Renal Artery , Renal Insufficiency, Chronic , Sodium Dodecyl Sulfate , Stem Cells , UreterABSTRACT
Secalonic acid D (SAD) could inhibit cell growth in not only sensitive cells but also multidrug resistant (MDR) cells. However, the molecular mechanisms need to be elucidated. Here, we identified that SAD possessed potent cytotoxicity in 3 pairs of MDR and their parental sensitive cells including S1-MI-80 and S1, H460/MX20 and H460, MCF-7/ADR and MCF-7 cells. Furthermore, SAD induced cell G2/M phase arrest the downregulation of cyclin B1 and the increase of CDC2 phosphorylation. Importantly, JNK pathway upregulated the expression of c-Jun in protein level and increased c-Jun phosphorylation induced by SAD, which was linked to cell apoptosis c-Jun/Src/STAT3 pathway. To investigate the mechanisms of upregulation of c-Jun protein by SAD, the mRNA expression level and degradation of c-Jun were examined. We found that SAD did not alter the mRNA level of c-Jun but inhibited its proteasome-dependent degradation. Taken together, these results implicate that SAD induces cancer cell death through c-Jun/Src/STAT3 signaling axis by inhibiting the proteasome-dependent degradation of c-Jun in both sensitive cells and ATP-binding cassette transporter sub-family G member 2 (ABCG2)-mediated MDR cells.
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To summarize the projects granted by National Natural Science Foundation of China related to tumor biomarkers in laboratory medicine from 2010 to 2018. The projects are categorized and analyzed according to tumor classification, biochemical properties of biomarkers. What′s more, characters of granted projects of tumor biomarkers in recent 9 years are summarized and development trends are pointed out.
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Regarding to the sharply increased applications and relatively lower rate of successful funding for major projects in the fields of Critical Care/Wounds and Injuries/Burns/Plastic Surgery from the National Natural Science Foundation of China in recent years, the author summarized the funded projects in this specific field from 2014 to 2018, discussed the characteristics and trends of these applications and grants, and summarized the hotspot issues and frontier researches so as to help the applicants in the future.
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To summarize the projects granted by National Natural Science Foundation of China related to tumor biomarkers in laboratory medicine from 2010 to 2018. The projects are categorized and analyzed according to tumor classification, biochemical properties of biomarkers. What's more, characters of granted projects of tumor biomarkers in recent 9 years are summarized and development trends are pointed out.
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Regarding to the sharply increased applications and relatively lower rate of successful funding for major projects in the fields of Critical Care/Wounds and Injuries/Burns/Plastic Surgery from the National Natural Science Foundation of China in recent years, the author summarized the funded projects in this specific field from 2014 to 2018, discussed the characteristics and trends of these applications and grants, and summarized the hotspot issues and frontier researches so as to help the applicants in the future.
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Urate oxidase (Uox), an enzyme catalyzing oxidation of uric acid to allantoin, is widely used as diagnostic reagents and for treatments of uarthritis and hyperuricemia diseases. In our study, a higher Uox producer, bacterial strain OUC-1, was isolated from soil samples. The 16S rRNA gene sequence of strain OUC-1 showed 99% identity to the homologous fragments of Bacillus fastidiosus. After purification, Uox showed the optimal pH and temperature was 10.0 and 40 °C. The Km value of Uox was (0.15±0.04) mmol/L (n=5) with uric acid as the substrate. Uox activity was enhanced by Mg²⁺, and seriously inhibited by Zn²⁺ and SDS. Then the uox gene of B. fastidiosus OUC-1 was amplified and sequenced. The 3D structures of Uox, predicted with SWISS-MODEL, showed a homotetramer structure with a subunit molecular weight of 35.38 kDa. Finally, the gene coding for the B. fastidiosus Uox was successfully cloned and heterologously expressed in E. coli, which provides theoretical basis and technical support for improvement of Uox in the future.
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Euphorbia factor L2, a lathyrane diterpenoid isolated from caper euphorbia seed (the seeds ofL.), has been traditionally applied to treat cancer. This article focuses on the cytotoxic activity of Euphorbia factor L2 against lung carcinoma A549 cells and the mechanism by which apoptosis is induced. We analyzed the cytotoxicity and related mechanism of Euphorbia factor L2 with an MTT assay, an annexin V-FITC/PI test, a colorimetric assay, and immunoblotting. Euphorbia factor L2 showed potent cytotoxicity to A549 cells. Euphorbia factor L2 led to an increase in reactive oxygen species (ROS) generation, a loss of mitochondrial electrochemical potential, release of cytochromeactivation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase, suggesting that Euphorbia factor L2 induced apoptosis through a mitochondrial pathway. The cytotoxic activity of Euphorbia factor L2 in A549 cells and the related mechanisms of apoptotic induction provide support for the further investigation of caper euphorbia seeds.
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Objective To study the apoptotic effect of brazilein on human lung cancer A549 cells and endoplasmic reticulum stress .Methods The cytotoxic activity was tested by MTT assay in A549 cells .The flow cytometry was used to detect apoptosis . Western blotting was performed to detect GRP78 and cyto c protein expression .Results The IC50 values of brazilein against A549 cells was (5 .36 ± 0 .62)μmol/L .After treatment with 0 ,5 ,10 and 20 μmol/L brazilein for 48 h ,the percent of apoptosis was (1 .15 ± 0 .32)% ,(19 .61 ± 4 .52)% ,(30 .18 ± 6 .35)% and (39 .48 ± 7 .44)% respectively .There was significant difference among the different treatment (P< 0 .05) .Compared with control group ,the protein expression of GRP78 and cytosolic cyto c was in‐creased after 5 ,10 and 20 μmol/L brazilein treated for 48 h .Conclusion Brazilein induced apoptosis in human lung cancer A549 cells though endoplasmic reticulum stress pathway .
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Objective To explore the feature of functional connectivity of default mode network (DMN) and salience network (SN) in unmedicated schizophrenia patients during a resting state by functional magnetic resonance imaging (fM-RI). Methods The SPM8 and DPARSFA softwares combined with independent component analysis (ICA) were used to in-vestigate functional connectivity (FC) of the DMN and SN in 27 unmedicated patients with schizophrenia and 27 age-and gender-matched healthy controls. Results Concerning the DMN, patients with schizophrenia showed decreased FC in right inferior frontal gyrus , right precuneus(unadjusted P<0.05)and increased FC in right middle cingulate gyrus, left middle frontal gyrus(unadjusted P<0.05). With regard to the SN, patients showed reduced connectivity in left inferior frontal gyrus, right inferior frontal gyrus, left anterior cingulate, left postcentral gyrus(unadjusted P<0.05)and increased connectivity in left superior temporal gyrus(unadjusted P<0.05). Correlation analyses showed that the increased FC of left superior temporal gyrus significantly correlated with PANSS-positive symptoms(r=0.568,P=0.002)and decreased FC of right precuneus significantly negatively correlated with delusion symptom(r=-0.458,P=0.016). Conclusion This study provides evidence for resting state functional abnormalities of DMN and SN in unmedicated schizophrenia patients. These aberrant function connectivities in some brain regions of the two networks could be a source of abnormal introspectively-oriented mental actives.
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Previous studies have demonstrated that the Chinese medicine paeoniflorin, derived from the Ranunculaceae plant peony, peony, purple peony root, was able to have anti-inflammatory, anti-ulcer, anti- hypersusceptibility and anti-oxidation activity. In order to elucidate the pesticide effect and the mechanisms by which paeoniflorin exerts its effect of anti-inflammation and immunoregulation on oxazolone-induced colitic mice, disease activity index (DAI) and histological grading of colitis (HGC) were evaluated in animal model. Moreover, the expressions of HBD-2, IL-6 and IL-10 of mice with experimental colitis were observed with immunohistochemistry and RT-PCR in this study. Results showed that DAI and HGC of oxazolone control group was significantly higher than that of normal control group, and that paeoniflorin groups and 5-ASA group, compared with oxazolone control group, could alleviate the symptoms and histological damages of colitic mice (P
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Previous studies have demonstrated that the Chinese medicine paeoniflorin, derived from the Ranunculaceae plant peony, peony, purple peony root, was able to have anti-inflammatory, anti-ulcer, anti-hypersusceptibility and anti-oxidation activity. In order to elucidate the pesticide effect and the mechanisms by which paeoniflorin exerts its effect of anti-inflammation and immunoregulation on oxazolone-induced colitic mice, disease activity index (DAI) and histological grading of colitis (HGC) were evaluated in animal model. Moreover, the expressions of HBD-2, IL-6 and IL-10 of mice with experimental colitis were observed with immunohistochemistry and RT-PCR in this study. Results showed that DAI and HGC of oxazolone control group was significantly higher than that of normal control group, and that paeoniflorin groups and 5-ASA group, compared with oxazolone control group, could alleviate the symptoms and histological damages of colitic mice (P < 0.05, P < 0.01). The expression of HBD-2 and IL-6 cytokine on the colon of colitic mice was higher than that of normal control, paeoniflorin and 5-ASA groups (P < 0.05, P < 0.01), but the expression of IL-10 is lower than that of normal control, paeoniflorin and 5-ASA groups (P < 0.05, P < 0.01). The positive correlations were demonstrated between the expression of (HBD-2 and IL-6) and DAI (Pearson r = 0.728, Pearson r = 0.758, P < 0.01, respectively), (HBD-2 and IL-6) and HGC (Pearson r = 0.819, Pearson r = 0.825, P < 0.01, respectively), whereas, the negative correlations were demonstrated between the expression of IL-10 and DAI (Pearson r = -0.789, P < 0.01), IL-10 and HGC (Pearson r = -0.725, P < 0.01). It can be concluded that to some extent paeoniflorin effectively alleviate the symptoms of oxazolone-induced colitis through regulating the expression of HBD-2, IL-6 and IL-10.
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AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.
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AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.
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Prostate cancer is a disease involving complicated multiple-gene alterations. Both NKX3.1 and p53 are related to prostate cancer and play crucial roles in prostate cancer progression. However, little is known about the relationships and interactions between p53 and NKX3.1 in prostate cancer. We found that NKX3.1 expression is down-regulated by over-expression of wild type (wt) p53 in prostate cancer LNCaP cells. NKX3.1 is down-regulated at both the mRNA and protein levels by p53 over- expression due to either transient transfection of exogenous p53 or induction of endogenous p53. p53 over-expression represses androgen-induced transactivation of NKX3.1 by inhibiting the promoter of the androgen acceptor (AR) gene and by blocking AR-DNA binding activity. In addition, transfection with the p21 expression vector (pPSA-p21) showed that p21 does not reduce NKX3.1 expression, indicating that NKX3.1 expression is not the result of nonspecific effects of cell growth arrest. Our results provide biochemical and cellular biologic evidence that NKX3.1 is down-regulated by p53 over-expression in prostate cancer cells.
Subject(s)
Male , Humans , Tumor Suppressor Protein p53/genetics , Transcription Factors/genetics , Transcriptional Activation/drug effects , Response Elements , RNA, Messenger/genetics , Prostatic Neoplasms/genetics , Promoter Regions, Genetic/genetics , Plasmids/genetics , Homeodomain Proteins/genetics , Genes, Reporter/genetics , Down-Regulation , Cell Line, Tumor , Androgens/pharmacologyABSTRACT
Objective To carry out a systematic study on the chemical constituents in the fruits of Momordica grosvenori. Methods To isolate pure compounds by using repeated column chromatography,while the structure of a new compound was determined by detailed spectral analysis. Results Four cucurbitane triterpenoid glycosides, mogroside Ⅱ E (Ⅰ), mogroside Ⅲ (Ⅱ), grosmomoside Ⅰ (Ⅲ), and mogroside Ⅴ (Ⅳ) were isolated from the 50% ethanolic extract of the fruits of M. grosvenori. Conclusion Grosmomoside Ⅰ is a new compound identified as mogrol-3-O-β-D-glucopyranoside-24-O-{[β-D-glucopyranosyl (2-1 ) ]- [β-D-glucopyranosyl (6-1 ) ]-β-D-galactopyranoside } and the other three compounds are known compounds.
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<p><b>OBJECTIVE</b>To evaluate the status of eight RHD specific exons in 131 Han Chinese blood donors who were classified as RhD-negative by serological methods and explore the genomic structure of RHD gene among the Han Chinese. The Rh blood group system has the highest prevalence of polymorphisms among human blood group systems and is clinically significant in transfusion medicine. The Rh antigens are expressed on polypeptides encoded by two highly homologous genes, RHD and RHCE. Recent molecular studies have shown that the RhD-negative trait could be generated by multiple genetic mechanisms and is ethnic group-dependent.</p><p><b>METHODS</b>The polymerase chain reaction using-sequence specific primers (PCR-SSP) was used to amplify exons 2, 3, 4, 5, 6, 7, 9 and 10 of RHD gene and exons 1, 2 and 5 of RHCE gene, as well as intron 4 in each of them.</p><p><b>RESULTS</b>The 131 cases of RhD-negative phenotypes consisted of 60 ccee, 58 Ccee, 5 ccEe, 5 CcEe and 3 CCee. Among them, 83 with the Rh ccee or ccEe phenotypes (63.4%) lacked the eight RHD exons indicated above, while 26 cases with the Rh Ccee, CCee, CcEe phenotypes (19.9%) had all the RHD exons examined. Twenty-two individuals with the Ccee, CCee, CcEe phenotypes (16.8%) carried at least one RHD exon. The phenotypes of the RhD negative individuals carrying the RHD gene were Rh CC or Cc, but not cc.</p><p><b>CONCLUSIONS</b>Three classes of RhD-negative polymorphisms among a population of Han Chinese were observed. Antigen association analysis suggested the existence of a novel class of RhD-negative associated haplotype in Han Chinese. This haplotype consisted of a normal RHCE allele and a nonfunctional RHD gene. It may be beneficial to redefine the RhD-negative blood group among Chinese populations upon clarification of the mechanisms of RHD gene expression and RhD antigen immunization.</p>
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Humans , Asian People , China , Ethnicity , Genetics , Phenotype , Polymerase Chain Reaction , Polymorphism, Genetic , Rh-Hr Blood-Group System , GeneticsABSTRACT
AIM: To observe the effect of curcumin on the proliferation and apoptosis of prostate cancer cell line LNCap.METHODS: LNCap cells were treated with different doses(10 ?mol/L,20 ?mol/L,30 ?mol/L,40 ?mol/L) of curcumin and its effects were analyzed in cell growth and apoptosis by microscope,MTT colorimetric assay and flow cytometry.The expression of prostate specific antigen(PSA) was measured by AXSYM~(TM) system-chemical luciferase methods and expression of androgen receptor (AR) was detected by Western blotting.RESULTS: The results showed that curcumin inhibited the proliferation of LNCap cells.The cell growth was inhibited by curcumin in a dose dependent manner and the optimal dose and time was 40 ?mol/L,24 h.Curcumin induced apoptosis in LNCap cells,the most dramatic dose was 40(?mol/L) curcumin,at this dose the apoptosis rate was 9.23%. Curcumin inhibited the expression of PSA in LNCap cells and the most dramatic dose and time was 40 ?mol/L,24 h.The PSA in this group was 20% of the control group.Curcumin inhibited the expression of AR on prostate cancer cells.CONCLUSION: Curcumin decreases proliferation and induces apoptosis in LNCap cells in a dose-dependent manner.Curcumin also inhibites the expression of PSA and AR on LNCap cells.