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Diabetes Mellitus , Sirtuina 3 , Humanos , Sirtuina 3/metabolismo , Oxidación-Reducción , AcetofenonasRESUMEN
OBJECTIVE@#To observe the effects of acupuncture on neurologic function and serum inflammatory factors in patients after thrombolysis in acute ischemic stroke (AIS).@*METHODS@#A total of 102 AIS patients with onset to treatment time (OTT) ≤3 h were randomly divided into an observation group and a control group, 51 cases each group. In the control group, thrombolysis and conventional medical treatment were applied. On the basis of the treatment as the control group, acupuncture at Shuigou (GV 26), Zhongwan (CV 12), Qihai (CV 6), Neiguan (PC 6), etc. was applied in the observation group, 30 min each time, once a day. Both groups were treated for 2 weeks. Before and after treatment, the scores of National Institutes of Health stroke scale (NIHSS), modified Rankin scale (mRS), modified Barthel index (MBI) and serum level of homocysteine (Hcy), hypersensitive C-reactive protein (hs-CRP) were compared, and the clinical efficacy was evaluated in the two groups.@*RESULTS@#After treatment, the scores of NIHSS, mRS and serum level of Hcy, hs-CRP were decreased compared with those before treatment (P<0.05), while the MBI scores were increased (P<0.05) in the two groups. The scores of NIHSS, mRS and serum level of Hcy, hs-CRP in the observation group were lower than those in the control group (P<0.05, P<0.01), the MBI score in the observation group was higher than that in the control group (P<0.01). The total effective rate was 88.2% (45/51) in the observation group, which was superior to 70.6% (36/51) in the control group (P<0.05).@*CONCLUSION@#Acupuncture could promote the recovery of neurologic function in patients after thrombolysis in AIS, improve the ability of daily living, which may be related to reducing the level of inflammatory factors, thus inhibiting inflammatory response and improving cerebral ischemia reperfusion injury.
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Humanos , Estados Unidos , Accidente Cerebrovascular Isquémico , Proteína C-Reactiva , Terapia por Acupuntura , Inflamación , Homocisteína , Hipersensibilidad , Terapia TrombolíticaRESUMEN
Liraglutide is an analog of glucagon⁃like peptide⁃1 and plays an important role in the treatment of diabetes. But the specific mechanism of liraglutide in improving the function of pancreatic β cells is still unclear. Therefore, high glucose (33 mmol/ L) was used to induce islet MIN6 cells for 48 hours to establish a high glucose injury model in this study. The CCK⁃8 assay was used to detect cell viability, and the results showed that the viability of MIN6 cells in the high glucose group decreased (P<0. 05) compared with the control group, and the cell viability increased after liraglutide treatment (P<0. 05). Using the mouse insulin detection kit and ATP content detection kit, we found that both the insulin re⁃ lease (P<0. 01) and ATP content decreased (P<0. 001) in the high glucose group compared with the control group, and after liraglutide treatment both insulin release (P < 0. 05) and ATP content (P < 0. 001) increased. We used the mitochondrial membrane channel pore (MPTP) fluorescence detection kit in MIN6 cells and found that the green fluorescence intensity of the high glucose group was significant⁃ ly decreased (P<0. 001) compared with the control group, and after liraglutide treatment the green fluo⁃ rescence intensity was significantly increased (P<0. 001). The DCFH⁃DA probe combined with flow cy⁃ tometry was used to detect the level of reactive oxygen species (ROS). We found that compared with the control group, the ROS level in the high glucose group was significantly increased (P<0. 001), and de⁃ creased by liraglutide treatment (P<0. 01). Intracellular malondialdehyde (MDA) contents, superoxide dismutase (SOD) and catalase (CAT), and lactate dehydrogenase (LDH) activities in the cell superna⁃ tant were measured, and the levels of MDA and LDH were significantly increased (P<0. 05), and the levels of SOD and CAT were significantly decreased (P<0. 01) in the high glucose group compared with the control group. After liraglutide treatment, the levels of MDA and LDH were decreased (P<0. 05), and the levels of SOD and CAT were increased (P<0. 05). The results of Western blotting showed that the expression of UCP2 was upregulated in the high glucose group (P<0. 01) compared with the control group, and after liraglutide treatment the expression of UCP2 decreased (P<0. 05). The results indica⁃ ted that liraglutide has significant effects in high⁃glucose induced mitochondrial damage, oxidative stress and insulin secretion in MIN6 cells, which will provide a theoretical basis for clinical utilization of liraglu⁃ tide.
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Objective: Peptidyl alkaloids, a series of important natural products can be assembled by fungal non-ribosomal peptide synthetases (NRPSs). However, many of the NRPSs associated gene clusters are silent under laboratory conditions, and the traditional chemical separation yields are low. In this study, we aim to discovery and efficiently prepare fungal peptidyl alkaloids assembled by fungal NRPSs. Methods: Bioinformatics analysis of gene cluster containing NRPSs from the genome of Penicillium thymicola, and heterologous expression of the putative gene cluster in Aspergillus nidulans were performed. Isolation, structural identification, and biological evaluation of the product from heterologous expression were carried out. Results: The putative tri-modular NRPS AncA was heterologous-expressed in A. nidulans to give anacine (1) with high yield, which showed moderate and selective cytotoxic activity against A549 cell line. Conclusion: Heterologous expression in A. nidulans is an efficient strategy for mining fungal peptidyl alkaloids.
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Endophytic fungi are promising producers of bioactive small molecules. Bioinformatic analysis of the genome of an endophytic fungus
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Objective: To explore the effects of 3-phosphate dependent protein kinase 1-protein kinase B (PDK1-Akt) signaling pathway on the transcription, expression and function of cardiac hyperpolarized activated cyclic nucleotide gated 4 (HCN4) ion channels. Methods: Atrial myocytes were obtained from healthy male wild-type C57 mice and heart-specific PDK1 knockout mice (PDK1-KO) by enzymolysis. Then the atrial myocytes were divided into blank control group and PDK1-KO group. In further studies, the isolated atrial myocytes were cultured and further divided into drug control group (treated with dimethyl sulfoxide (DMSO)) and PDK1 knockdown group (treated with 1 μg/ml PDK1 short hairpin RNA (shRNA) interference plasmid), SC79 group (treated with 8 μmol/ml SC79), GSK2334470 group (treated with 10 nmol/L GSK2334470) and PDK1 knockdown+SC79 group (8 μmol/ml SC79 and 1 μg/ml PDK1 shRNA interference plasmid). Real time quantitative PCR (qRT-PCR) was used to detect the mRNA expression levels of PDK1 and HCN4, Western blot was used to detect the protein expression levels of PDK1, Akt and HCN4, the whole cell patch clamp was used to detecte the current density of HCN, and immunofluorescence was used to detecte the expression of HCN4 protein on atrial cells. Results: (1) the expression levels of HCN4 mRNA (1.46±0.03 vs. 0.99±0.01, P<0.001) and protein (1.14±0.02 vs. 1.00±0.06, P=0.017) in PDK1-KO group were higher than those in blank control group. The HCN current density in PDK1-KO group was higher than that in blank control group((-17.47±2.00) pA/pF vs. (-12.15±2.25) pA/pF, P=0.038). (2) The functions of PDK1 shRNA and specific Akt agonist SC79 were verified by comparing the PDK1 knockdown group and SC79 group with the drug control group. The results showed that the expression levels of PDK1 mRNA and protein in PDK1 knockdown group were lower than those in drug control group, and the expression level of phosphorylated Akt (Thr 308) protein in SC79 group was higher than that in drug control group. (3) The expression levels of HCN4 mRNA (3.61±0.46 vs. 1.00±0.08, P<0.001) and protein (2.33±0.11 vs. 1.00±0.05, P<0.001) in GSK2334470 group were higher than those in drug control group. (4) To reduce the effect of drug-miss target, the cultured atrial myocytes were transfected with shRNA plasmid of PDK1 and intervened with SC79. The results showed that the expression of HCN4 mRNA in PDK1 knockdown group was higher than that in the drug control group (1.76±0.11 vs. 1.00±0.06, P<0.001), and PDK1 knockdown+SC79 group (1.76±0.11 vs. 1.33±0.07, P=0.003). In PDK1 knockdown+SC79 group, the mRNA expression level was also higher than that in the drug control group (1.33±0.07 vs. 1.00±0.06, P<0.001). The expression level of HCN4 protein in PDK1 knockdown group was higher than that in drug control group (1.15±0.04 vs. 1.00±0.05, P=0.003). As for the The expression level of HCN4 protein, there was no significantly statistical difference between the PDK1 knockdown+SC79 group and the drug control group (P>0.05), but PDK1 knockdown+SC79 group was lower than PDK1 knockdown group (0.95±0.01 vs. 1.15±0.04, P<0.001). In patch clamp experiments, the results showed that the HCN current density was (-13.27±1.28) pA/pF in the drug control group, (-18.76±2.03) pA/pF in the PDK1 knockdown group, (-13.50±2.58) pA/pF in the PDK1 knockdown+SC79 group; the HCN current density of PDK1 knockdown group was higher than that of drug control group (P<0.001), but there was no significant difference between PDK1 knockdown+SC79 group and drug control group (P>0.05). (5) The results of immunofluorescence showed that the brightness of green fluorescence of PDK1 knockdown group was higher than that of drug control group, indicating that the expression of HCN4 localized on cell membrane was increased. However, the green fluorescence of PDK1 knockdown+SC79 group was lighter than that of PDK1 knockdown group, suggesting that the expression of HCN4 in PDK1-knockdown cell membrane decreased after further activating Akt. Conclusion: PDK1-Akt signaling pathway is involved in the regulation of HCN4 ion channel transcription, expression and function.
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Animales , Masculino , Ratones , Canales Catiónicos Regulados por Nucleótidos Cíclicos , Canales Regulados por Nucleótidos Cíclicos Activados por Hiperpolarización/metabolismo , Miocitos Cardíacos/metabolismo , Canales de Potasio/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de SeñalRESUMEN
italic>Shiraia bambusiccola is an important medicinal fungus in China. Hypocrellins with perylenequinone skeleton are main bioactive components of Shiraia bambusiccola, which are widely used in food, medicine, pesticide and other fields as natural photosensitizers. For example, "hypocrellin ointment" has already been used clinically. As a rare and vulnerable species, wild Shiraia bambusiccola resources are very limited. Due to the complex structure and chanllenge in chemical total synthesis of hypocrellins, it is urgent to find an effective strategy to rationally utilize its medicinal value while protecting the wild resources. In this study, a candidate gene cluster hpc was identified in Shiraia sp. cfcc 84681 based on careful bioinformatic analysis. A heterologous expression system for hpc gene cluster was successfully constructed and a mutant strain with high yield of hypocrellins was obtained, which mainly produced hypocrellin A and isohypocrellin A. The main ingredients in the mutant strain are consistent with that in the wild Shiraia bambusiccola. These results provide a new strategy to solve the shortage of wild Shiraia bambusiccola resources.
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Aim To explore the effects of parthenolide (PTL) on apoptosis, invasion and migration of NSCLC cell line H1975 as well as the possible mechanism. Methods MTT assay and colony formation assay were used to measure cell proliferation. Flow cytometry with Annexin V-FITC/PI double staining were employed to measure cell apoptosis. Transwell assay was applied to measure cell invasion and migration. The expression of apoptosis-related proteins, invasion and migration-associated proteins and PBK/Akt signaling pathway-related proteins were detected using Western blot. Results MTT assay and colony formation assay results showed that the proliferation of HI975 cells was significantly inhibited with the increase of PTL concentration, and compared with control group, the differences were statistically significant (P < 0. 05). Annexin V-FITC/PI double staining result indicated that PTL induced apoptosis in HI975 cells (P <0. 01). The results of transwell assay demonstrated that PTL significantly inhibited the invasion and migration of HI975 cells (P <0. 01). Western blot analysis revealed that PTL increased the expression of Bax and reduced the expression of Bcl-2, HIF-1 a, MMP-9, Akt and p-Akt (Ser473) in HI975 cells ( P < 0. 05 ), meanwhile, the cleavage of caspase-3 was detected. Conclusions PTL can significantly induce apoptosis and inhibit the invasion and migration of HI975 cells, and the mechanism may be related to the inhibition of PI3K/Akt signaling pathway.
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Six novel monacolin analogs, monacolins V-V (1-6), together with seven known ones (7-13), were isolated from the ethyl acetate extract of red yeast rice. Their structures and absolute configurations were determined by spectroscopic methods, especially 2D NMR (H-HCOSY, HSQC, HMBC, and NOESY/ROESY) and CD spectroscopic analyses as well as chemical derivation. Monacolins V (2) and V (3) represent the first examples of monacolins with 3-hydroxybutyrate substitute. The anti-inflammatory inhibitory activities against the lipopolysaccharide (LPS) induced NO production in BV-2 cells as well as antioxidant activities against rat liver microsomal lipid peroxidation were evaluated.
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OBJECTIVE@#Epstein-Barr virus associated gastric cancer (EBVaGC) is different from the traditional gastric cancer (Epstein-Barr virus non-associated gastric cancer, EBVnGC), and has unique clinicopathological features. This study investigated the largest single center cancer series so as to establish the clinicopathological and molecular characteristics of EBVaGC in China.@*METHODS@#A retrospective analysis was conducted on EBVaGC and EBVnGC patients diagnosed at Peking University Cancer Hospital from 2003 to 2018 by comparing their clinicopathological features and prognosis. The gastric cancer (GC) dataset of public database was analyzed to obtain differentially expressed genes. The expression of important genes and their association with prognosis of GC were verified in GC tissues from our hospital.@*RESULTS@#In this study, 3 241 GC patients were included, and a total of 163 EBVaGC (5.0%) patients were identified. Compared with EBVnGC, EBVaGC was higher in male and younger patients, and positively associated with remnant GC, poorly differentiated adenocarcinoma, and mixed type GC. EBVaGC was inversely related to lymph node metastasis. The 5-year survival rate of EBVnGC and EBVaGC was 59.6% and 63.2% respectively (P<0.05). In order to explore molecular features of EBVaGC, the Cancer Genome Atlas (TCGA) dataset was analyzed (n=240), and 7 404 significant differentially expressed genes were obtained, involving cell proliferation, apoptosis, invasion and metastasis. The down-regulated invasion/metastasis gene SALL4 and the up-regulated immune checkpoint gene PD-L1 were important molecular features of EBVaGC. Validation of these two genes in large GC series showed that the majority of the EBVaGC was SALL4 negative (1/92, 1.1%, lower than EBVnGC, 303/1 727, 17.5%), and that PD-L1 was mostly positive in EBVaGC (81/110, 73.6%, higher than EBVnGC, 649/2 350, 27.6%). GC patients with SALL4 negative and PD-L1 positive were often associated with better prognosis.@*CONCLUSION@#EBVaGC is a unique subtype of GC with less metastasis and a good prognosis. It also has a distinct molecular background. The down-regulation of invasion/metastasis gene SALL4 and up-regulation of immune checkpoint gene PD-L1 are important molecular features.
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Femenino , Humanos , Masculino , China , Infecciones por Virus de Epstein-Barr/complicaciones , Herpesvirus Humano 4 , Estudios Retrospectivos , Neoplasias Gástricas/etiologíaRESUMEN
Objective To determine whether urinary myeloperoxidase to creatinine ratio (MCR) can serve as a marker for diagnosis of urinary tract infection (UTI).Methods Patients suspected of UTI were consecutively enrolled and further divided into the culture positive and the sterile groups according to urine culture results. Subsequently, MCR, white blood cell (WBC) and bacteria in the urinary samples from patients were detected and compared between the two groups.Results Finally, 253 patients were enrolled including 157 urine culture positive patients and 96 urine culture negative patients (sterile group). After logarithmic transformation in 2 as the base, the MCR, WBC, and bacteria were separately presented as log, log(quantitative) , and log. The values of log(8.6±2.5 vs. 5.4±1.5, t=-12.453, P=0.001), log(quantitative) (8.0±2.5 vs. 5.2±1.8, t=-10.332, P=0.001), log (11.4±2.5 vs. 8.2±2.8, t=-9.297, P=0.001) and WBC (semi-quantitative) [2 (interquartile range 1, 3) vs. 1 (interquartile range 0.5, 1), Z=-7.580, P=0.001] showed significant difference between the urine culture positive group and the sterile group. Among the urine culture positive group, the values of log of the gram positive and gram negative subgroups were 7.2±2.5 and 9.0±2.4 (t=4.016, P=0.001), respectively. The correlation between log and log (quantitative), log, WBC (semi-quantitative) was 0.708 (Pearson correlation, P=0.001), 0.381 (Pearson correlation, P=0.001), and 0.606 (Spearman correlation, P=0.001), respectively. Conclusions MCR is positively correlated with WBC counts and could be served as a promising biomarker for diagnosis of UTI. MCR could be even used for initial inference of infectious bacteria types of UTI.
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Fungal genomes carry many gene clusters seemingly capable of natural products biosynthesis, yet most clusters remain cryptic or down-regulated. Genome mining revealed an unconventional paraherquonin-like meroterpenoid biosynthetic gene cluster in the chromosome of . The cryptic or down-regulated pathway was activated by constitutive expression of pathway-specific regulator gene encoded within biosynthetic gene cluster. Chemical analysis of mutant -OE: extracts enabled the isolation of four berkeleyacetal congeners, in which two of them are new. On the basis of careful bioinformatic analysis of the coding enzymes in the gene cluster, the biosynthetic pathway of berkeleyacetals was proposed. These results indicate that this approach would be valuable for discovery of novel natural products and will accelerate the exploitation of prodigious natural products in filamentous fungi.
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Objective To compare the effect of ARIMA models with and without covariates in predicting the number of HIV infections among young students in Dalian. Methods First, univariate correlation analysis was performed on the network, STD sequence and HIV sequence to understand whether there was a correlation and lag relationship between them. Secondly, variables with the strongest correlation and predictive value and HIV infection numbers were used as the baseline data to establish an ARIMA model with covariates and a general ARIMA model without covariates, and to predict the HIV number from 2019 to 2021. The average absolute errors were used as evaluation indexes to compare the prediction effects of the two models. Results A total of 841 cases of HIV infection among young students were reported in Dalian from 2013 to 2021. The results of univariate correlation analysis showed that the search index of the keyword AIDS in the Baidu Index in a given month from 2013 to 2019 was significantly positively correlated with the number of HIV infections in that month (r=0.302, P=0.006), and gonorrhea was negatively correlated with the number of HIV infections with a lag of 2 months (r=-0.250, P =0.024). Using gonorrhea incidence number and HIV infection number as the basic data, an ARIMA model with covariates and a general ARIMA model without covariates were established to predict the number of HIV infection among young students from 2019 to 2021, and the average absolute errors were 17.621% and 66.17%, respectively. Conclusion Compared with the general ARIMA model without covariates, the ARIMA model based on the combined use of STD incidence and HIV infection is more suitable for predicting the number of HIV infections among young students in Dalian, but the average absolute error of the model is still large, which needs further improvement in the future research.
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OBJECTIVE@#To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro.@*METHODS@#placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4 T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4CD25Foxp3Treg in T lymphocytes was accounted.@*RESULTS@#After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4 T lymphocytes were cultured for 48 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (<0.01).@*CONCLUSIONS@#Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4CD25Foxp3Treg in vitro.
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Femenino , Humanos , Embarazo , Factores de Transcripción Forkhead , Antígenos HLA-G , Células Madre Mesenquimatosas , Placenta , Linfocitos T ReguladoresRESUMEN
Epigenetic modifications have been proved to be a powerful way to activate silent gene clusters and lead to diverse secondary metabolites in fungi. Previously, inactivation of a histone H3 deacetylase in had led to pleiotropic activation and overexpression of more than 75% of the biosynthetic genes and isolation of ten compounds. Further investigation of the crude extract of strain resulted in the isolation of twelve new diterpenoids including three cassanes (-), one cleistanthane (), six pimaranes (-), and two isopimaranes ( and ) along with two know cleistanthane analogues. Their structures were elucidated by extensive NMR spectroscopic data analysis. Compounds and showed potent inhibitory effects on the expression of MMP1 and MMP2 (matrix metalloproteinases family) in human breast cancer (MCF-7) cells.
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Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.
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AIM:To investigate the effects of vinpocetine on inflammation of brain in intracerebral hemorrhage ( ICH) rats and to explore the underlying mechanisms .METHODS:All rats were randomly divided into sham group , ICH group, ICH with low dose of vinpocetine treatment group , ICH with medium dose of vinpocetine treatment group , and ICH with high dose of vinpocetine treatment group .Except sham group , the rats in other groups were injected with type VII col-lagenase to establish ICH model , and then the rats in vinpocetine treatment groups were received vinpocetine at 0.5, 1.0 or 1.5 mg/kg by intraperitoneal injection once a day for 7 days.After corresponding treatment , the impairment of neurological function in the rats was scored and the water content of the brain tissues was measured .Moreover, the activity of myeloper-oxidase (MPO) was determined by ELISA, and the protein expression of Toll-like receptors 4 (TLR4), nuclear factor-κB (NF-κB), intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molcule-1 (VCAM-1) in the brain tissues was determined by Western blot .RESULTS: Compared with ICH group , vinpocetine treatment significantly de-creased the scores of the impairment of neurological function and the water content of the brain tissues .Moreover, the activ-ity of MPO and the protein expression of TLR4, NF-κB, ICAM-1 and VCAM-1 were also reduced after vinpocetine treat-ment (P<0.05).CONCLUSION:Vinpocetine improves neurological function in ICH rats via suppression of inflamma -tion by inhibiting NF-κB signaling and expression of ICAM-1 and VCAM-1.
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Objective To explore the characteristics and biological functions of epigenetic treatment to DNMT1 expression in NSCLC cell lines. Methods Lung cancer cells were divided into 4 groups , and drugs with different concentrations. The effect of epigenetic treatment on NSCLC cell line was detected by CCK-8 and MTT. The effect of epigenetic treatment on the expression of DNMT1 in NSCLC cell line was detected by Western blot. Results CCK-8 and MTT assay showed that treatment with 5-Aza-CdR and MS-275 inhibited the proliferation of NSCLC cells. Western blot showed that treatment with 5-Aza-CdR and MS-275 inhibited the DNMT1 expression in NSCLC cells. Conclusion The expression of DNMT1 gene in the NSCLC cell lines may be suppressed effectively by epigenetic treatment , and the gene may inhibit the proliferation and growth of NSCLC cell lines.
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Objective To explore the effects of co‐cultured heat‐treated candida glabrata with rat tracheal epithelial (RTE) cells on the expression of Dectin‐1 and the production of IL‐6 and TNF‐α.Methods RTE cells in vitro were co‐cultured with heat‐treated candida glabrata bacteria liquid for 2 ,4 ,6 h ,while without co‐cultured RTE cells were used as control group .We observed the morphological changes of RTE cells ,detected the protein expressions of Dectin‐1 by Western blot ,used real‐time PCR to detecte the mRNA expressions of IL‐6 and TNF‐αand measured protein expression of IL‐6 and TNF‐αby ELISA .Results With the pass‐ing of time ,the RTE cells were damaged extensively and the expression of Dectin‐1 ,IL‐6 and TNF‐αbecame more and more signifi‐cant .Obviously ,there had significant difference in the expression of Dectin‐1 ,IL‐6 and TNF‐α between the co‐cultured 2 h group and the control group ,the co‐cultured 4 h group and the co‐cultured 2 h group ,the co‐cultured 6 h group and the co‐cultured 4 h group (P<0 .05) .Conclusion RTE cells have natural immune function .The Dectin‐1 involves in the recognition of heat‐treated Candida glabrata ,activating secretion of IL‐6 and TNF‐αand mediating inflammatory reaction .IL‐6 plays a negative regulation role .
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Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.