Table of Content

05 July 2019, Volume 39 Issue 7

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Cellular uptake of dandelion decoctosome is mediated by macropinocytosis

2019, 39(7):  925-931. 

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Objective To characterize the mechanism of cellular uptake of dandelion decoctosome. Methods Internalization of fluorescently-labeled dandelion decoctosome was evaluated in A549 cells. By using a range of chemical inhibitors to block specific pathways as well as RNA interference (RNAi) to knockdown key proteins involved in individual endocytic pathways, cellular uptake of dandelion decoctosome was assessed using flow cytometry and confocal microscopy. Results Lysosomotropic agents significantly decreased dandelion decoctosome entering into cells（P<0.01）. Moreover, inhibitors of macropinocytosis, including cytochalasin D and amiloride significantly reduced the internalization of dandelion decoctosome in a dose dependent manner（P<0.001）. Consistently, knockdowning of Rac1 and PAK1 by RNAi impeded the cell entry of dandelion decoctosome（P<0.05）. Conclusions dandelion decoctosome enters cells via endocytosis and predominantly via macropinocytosis.

Construction of prokaryotic expression vector, optimization of expression conditions and soluble expression of human TMEM39A

2019, 39(7):  932-936. 

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Objective To construct prokaryotic expression vector of human TMEM39A, optimize expression conditions and obtain soluble TMEM39A. Methods TMEM39A gene was amplified from HEK293 cells total RNA by RT-PCR to construct its prokaryotic expression vector pGEX-6P-1-TMEM39A, and induced expression was conducted at different temperatures, IPTG concentrations, A600nm values and time conditions. Finally, expression of the recombinant protein GST-TMEM39A abundantly were carried out to analyze its solubility and antigenicity by using the best induction conditions. Results The nucleotide sequence homology of the recombinant expression vector pGEX-6P-1-TMEM39A and TMEM39A in GenBank (BC021277.2) was 99.9 % and the amino acid sequence homology was 100 %. The recombinant protein GST-TMEM39A showed two specific bands at 69 ku and 60 ku by Western blot analysis. The optimal induction conditions for GST-TMEM39A was 25 ℃, A600nm value of 0.6, 0.05 mmol/L IPTG induced 6 h. On this basis, the fusion expression of TMEM39A and GST protein was obtained in a soluble form. Conclusions The prokaryotic expression vector of TMEM39A is successfully constructed, the optimal expression condition of GST-TMEM39A is determined and its soluble expression is realized, which lay a material foundation for the subsequent purification of TMEM39A and the preparation of antibodies for functional studies, and provide scientific basis for further study about the relationship between TMEM39A and many diseases or viruses.

lncRNA expression profile analysis in hypotrophic cardiomyocytes induced by high glucose

2019, 39(7):  937-942. 

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Objective To explore the long-chain noncoding RNA (lncRNAs) expression and its possible biological functional analysis in hypotrophic cardiomyocytes induced by high glucose. Methods Neonatal mice cardiomyocytes were cultured by high glucose for processing 48 h and then randomly divided into the control group (Con) and the high sugar group (HG), the long-chain noncoding RNA (lncRNAs) expression were measured by using high throughput sequencing. Results 14 differentially expressed lncRNAs were screened, of which 5 were upregulated (ENSMUST00000195674, ENSMUST00000183707, ENSMUST00000195992, MSTRG.8385.2, MSTRG.9749.1) and 9 downregulated (ENSMUST00000195426, ENSMUST00000186944, ENSMUST00000182639, ENSMUST00000181094, ENSMUST00000125001, ENSMUST00000210380, MSTRG. 8302.1, MSTRG.11564.1, MSTRG.13750.1). ENSMUST00000182639, ENSMUST00000181094, ENSMUST00000195674, MSTRG.8302.1 may participate in PI3K-Akt signaling pathway, AMPK signaling pathway, mTOR signaling pathway, Rap1 signaling pathway, HIF-1 signaling pathway, FoxO signaling pathway, Hypertrophic cardiomyopathy, cardiomyopathy, or Oxidative phosphorylation by regulating Igf1.Conclusions LncRNAs in hypertrophic cardiomyocytes induced by high glucose are differentially expressed and might participated in the pathophysiological processes of hyperglycemic cardiomyocyte hypertrophy through corresponding signaling pathways.

Minocycline alleviates LPS-induced neuroinflammation by increasing the expression of ΔFosB in emotion-regulating brain regions in mice

2019, 39(7):  943-948. 

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Objective To investigate the role of ΔFosB as a transcription factor in the reduction of lipopolysaccharide (LPS)-induced neuroinflammation by minocycline. Methods Adult C57BL/6 mice at 10 weeks of age received intraperitoneal (i.p.) injections of saline or minocycline (50 mg/kg) for three consecutive days. On the third day, mice were also injected via i.p. with saline or Escherichia coli LPS (0.83 mg/kg). The morphology of microglia indicating the state of neuroinflammation and ΔFosB as a transcription factor were examined by immunohistochemistry. Images of the staining were analyzed with Image-Pro Premier 3D. Statistical analysis was performed with GraphPad Prism. Results Minocycline alleviated LPS-induced neuroinflammation as evidenced by the reduced activation of microglia in locus coeruleus (LC). Minocycline significantly increased the number of ΔFosB-positive neurons in LC, the lateral parabrachial nucleus (LPB) and paraventricular nucleus of the hypothalamus (PVN). Conclusions Minocycline alleviates LPS-induced neuroinflammation possibly by increasing the expression of ΔFosB in emotion-regulating brain regions including LC, LPB and PVN.

PRMT1 regulates the expression of c-myc through C/EBPα in breast cancer cell line MDA-MB-231

2019, 39(7):  949-953. 

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Objective To investigate how PRMT1 regulates c-myc in breast cancer cells. Methods The expression of c-myc was detected in protein and mRNA levles in overexpressing C/EBPα and knocking down PRMT1 of MDA-MB-231 cells. ChIP-qPCR and EMSA were performed to detect the binding ability of C/EBPα to the promoter of c-myc. The promoter of c-myc activity regulated by PRMT1 and C/EBPα was detected by dual-luciferase report system. Results The expression of c-myc declined dramatically after overexpressing C/EBPα and knocking down PRMT1. The promoter of c-myc activity repressed by C/EBPα was weakened obviously after the methylation sites of C/EBPα at R35F, R156F and R165F were mutated. The regulation of c-myc by PRMT1 has a little effect after the methylation sites of C/EBPα at R35K, R156K and R165K were mutated. Conclusions PRMT1 regulates the transcription of c-myc by methylation of C/EBPα.

Optimization of differentiation protocols of directing human adipose derived mesenchymal stem cells into definitive endoderm cells

2019, 39(7):  954-960. 

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Objective To establish and optimize the induction system for differentiation of human adipose-derived mesenchymal stem cells (hADSCs) into definitive endodermal (DE) cells. Methods Based on a well-developed protocol inducing ESCs/iPS towards DEs，the effects of activin A, WNT activators, BMP4 and bFGF, which are essential for inducing of ESCs/iPS to DE, were evaluated during the differentiation of hADSCs towards DEs. RT-qPCR was used to detect mRNA levels of DE marker genes FOXA2 and SOX17. Western blot was used to measure protein levels of FOXA2 and SOX17. Immunofluorescence staining was used to identify the expression of FOXA2 and SOX17, which demonstrated the feature of DE cells. Results Low concentration of activin A was more suitable to induce the expression of DE markers than high concentration (p<0.001). The expression of FOXA2 and SOX17 were higher in differentiation medium supplemented with Chir99021 than Wnt3a (p<0.01). BMP4 and bFGF showed no or harmful effects in the differentiation of hADSC to DE (p<0.001). Conclusions Different pathways are needed during the DE differentiation from hADSC compared with ESCs/iPS.

Tectoridin and aucubin inhibit LPS-induced inflammatory response of human adipose mesenchymal stem cells

2019, 39(7):  961-966. 

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Objective To study the effects of tectoridin and aucubin on the immune regulation of human adipose mesenchymal stem cells (hAMSCs) induced by lipopolysaccharide (LPS).Methods CCK-8 cell proliferation test and cell cycle test were used to screen the appropriate concentration of drugs; LPS concentration was screened to establish an inflammatory response model; ELISA and RT-q PCR were used to detect the effects of both on the secretion of pro-inflammatory and anti-inflammatory factors by hAMSCs after LPS induction. Results 50μg/mL tectoridin and 100μg/mL aucubin could significantly promote the proliferation of normal hAMSCs (P < 0.05 ，P＜0.01). Both of them promoted the proliferation of hAMSCs induced by LPS (P<0.001), and inhibited the secretion of IL-6，TNF-α，IFN-γ (P＜0.05 ，P＜0.01) .Conclusions Tectoridin and aucubin could significantly inhibit LPS-induced hAMSCs inflammatory response.

Na+/H+ exchanger regulatory factor 1 is a potential predictor for clinical prognosis of colorectal adenocarcinoma

2019, 39(7):  967-972. 

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Objective To find factors related to colorectal adenocarcinoma invasion and migration. Methods The data of colorectal adenocarcinoma in TCGA and GEO databases were collected for SAM differential analysis. The differentially expressed genes were collected through DAVID, and genes related to invasion and migration were identified. In addition, cancer and paired adjacent tissues from 50 cases of Chinese colorectal adenocarcinoma patients with detailed clinical and prognostic information were collected for immunohistochemical staining to verify the above results. The ROC curve was drawn to evaluate the ability of this gene or its encoded protein for colorectal adenocarcinoma diagnosis. Results Totally 6 clusters of migration-related genes were found by DAVID analysis (p<0.05). The down-regulated genes were DLC1, EPB41L3, KIT, PARVA, SLC9A3R1 and TPM1. NHERF1, encoded by SLC9A3R1, is a scaffold protein that interacts with cytoskeleton and various signaling proteins. Data showed that both mRNA and protein levels of NHERF 1 in colorectal adenocarcinoma were lower than those in normal tissue, and its expression was decreased in high-grade and lymph node metastasis. Conclusions The decreased expression of NHERF1 is an independent factor associated with poorer survival of colorectal adenocarcinoma patients, which may result from its metastasis. Decreased expression of NHERF1 is a potential indicator for prediction of colorectal adenocarcinoma prognosis.

miR-17-5p induces podocytes apoptosis and inhibits glomerular epithelial protein 1 in mice

2019, 39(7):  973-977. 

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Objective To explore the role and mechanisms of miR-17-5p in the pathogenesis of childhood nephrotic syndrome. Methods miR-17-5p mimic were transfected into podocytes in kidney of mice (MPC5) for 24 h to harvest the cells. the potential targets of MiR-117-5P were searched using the prediction programs microRNA.org, TargetScan and PicTar. RT-PCR and Western blot were used to detect expression of PTPRO mRNA and protein. Ca2+ fluorescence indicator with Fluo-3-Amspecificity were used to measure intracellular free calcium concentration ([Ca2+]i) in podocytes. Annexin V/PI flow cytomeutry was used to test apoptosis rate of podocytes. Results According to the bioinformatics technology prediction, miR-17-5p derived from human or mouse all can bind to multiple sites of the corresponding PTPRO-3 'UTR region and inhibit the expression of PTPRO protein and mRNA. PTPRO overexpression inhibits miR-17-5p-induced increase of [Ca2+]i in podocytes in kidney of mice. PTPRO overexpression alleviates miR-17-5p-induced apoptosis of podocytes in kidney of mice. Conclusion miR-17-5p inhibits expression of PTPRO in podocytes in kidney of mice and stimulates increase of intracellular free calcium concentration ([Ca2+]i) in podocytes, inducing apoptosis of podocytes in kidney of mice.

Naringin inhibits hypoxia/reoxygenation-induced of rat cardiomyocyte cell line H9c2 injury

2019, 39(7):  978-982. 

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Objective To study the effect of naringin (NAR) on endoplasmic reticulum (ER) stress in cardiomyocytes injured by H9c2 hypoxia/reoxygenation (H/R).Methods H9c2 cardiomyocytes were randomly divided into three groups: control group, hypoxia/ reoxygenation group (H/R) for 24 h after hypoxia for 4 h, naringin 10, 20 and 40 μg /mL groups.Naringin was cultured 6 h before hypoxia for 4 h and then reoxygenation for 24 h. TUNEL detects apoptosis of cardiomyocytes after the experiment. Western blot was used to detect the expression of GRP78,ATF6,Bax and Bcl-2.Results Compared with control group, the cell apoptosis was increased, the expression of GRP78, ATF6, Bax protein expression and Bax/Bcl-2 were remarkably up-regulated (P < 0.05), but Bcl-2 protein expression was remarkably down-regulated in H/R group. Compared with the H/R group, the cell apoptosis decreased; the expression of GRP78, ATF6 , Bax and Bax/Bcl-2 were down-regulated (P < 0.05), but Bcl-2 was up-regulated in the groups with naringin three groups,especially naringin 20 and 40μg/mL groups had remarkably effect.Conclusion:The pretreatment of naringin can reduce the apoptosis of myocardial cell. The mechanism may be related to the inhibition of the apoptotic pathway of endoplasmic reticulum stress.

Cystathionine β synthase promotes autophagy to alleviate Alzheimer's disease through ATG5 sulfhydration

2019, 39(7):  983-988. 

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Objective To explore the effect of CBS on Alzheimer's disease and its mechanism. Methods Western blot was used to detect the over-expression of CBS and its effect on amyloid precursor protein (APP), LC3 and P62. The effect of CBS on Aβ1-42 in cell supernatant was detected by ELISA. The formation of H2S was detected by methylene blue method; The sulfhydration of ATG5 by H2S was verified by biotin conversion experiment and Western blot experiment. CBS was overexpressed in mice by injection of plasmid into the lateral ventricle; Y-maze test was used to verify the effect of CBS on learning and memory ability in mice. Results CBS decreased the level of Aβ1-42 in the cell supernatant (P<0.05) and inhibited the expression of APP (P<0.05); CBS promoted the formation of H2S (P<0.01); CBS promotes sulfhydration of ATG5; CBS promoted the expression of LC3 and inhibited the expression of P62, CBS promoted the level of autophagy; CBS promotes learning and memory ability of Alzheimer's disease model mice (P<0.05). Conclusions CBS promotes the sulfhydration of cysteine at position 19 of ATG5 by promoting the formation of hydrogen sulfide, thereby promoting autophagy and alleviating Alzheimer's disease.

Change and significance of CD8+CD25+Foxp3+Treg cells in peripheral blood of patients with type 2 diabetes

2019, 39(7):  989-994. 

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Objective To investigate the role of CD8+CD25+Foxp3+Treg（CD8+Treg）cells in the pathogenesis of type 2 diabetes (T2DM). Methods 90 patients with T2DM were selected and divided into NMAU group containing 45 T2DM patients (MAL<30 mg/24 h) and MAU group containing 45 patients with microalbuminuria (MAL 30~300 mg/24 h), based on urinary microalbumin (MAL). Additionally, 45 healthy patients were selected as control group (HC). The concentration of mAlb were measured by Specific protein analyzer; the proportion of CD8+Tregs were measured by flow cytometry; and the concentration of serum interleukin (IL)-8, 1β, 6, and tumor necrosis factor (TNF)-α were measured by Luminex200. Results Compared with the HC group [4.43 (3.26-5.75) %], the proportion of CD8+Tregs in the NMAU group [4.06 (3.15-5.43) %] and the MAU group [3.52 (2.30-4.87) %] were decreased, and lower in MAU group (P<0.05). The serum levels of TNF-α, IL-8, IL-6 and IL-1β in the MAU group and the NMAU group were higher than that of HC group (P<0.05). The percentage of CD8+Treg cells showed a negative correlation with TNF-α (r=-0.4448 P<0.01), IL-8 (r=-0.3412 P<0.05), IL-6 (r=-0.3653 P<0.05) and IL-1β (r=-0.3192 P<0.05) respectively; Logistic regression analysis showed that CD8+Tregs may have a protective effect on T2DM. Conclusion By regulating the proportion of peripheral blood CD8+ Tregs in patients with T2DM, it is possible to slow down the progression of diabetes.

Expression of miR-16-5p and its target mRNA in peripheral blood leukocytes of rhesus mulatta

2019, 39(7):  995-1000. 

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Objective To clarify the correlation between microRNA and aging in rhesus mulatta. Methods The monkeys were divided into four different age groups according to human aging, which were named the infant, the adult, the old and the longevity. Real-time PCR was used to detect the expression of microRNA and VEGFA, GHR, TBP, INSR, CLOCK, IKBKB, PIK3R1, LRP2, SSTR3, IGF1R, BCL2, PPM1D, IRS1, MAPK8, ADCY5, APP, NFE2L1, BDNF, PEX5 in peripheral blood leukocytes. Using microRNA target genes forecast website miRwalk, TargetSan, miRecords, miRDB to predicte miR-16-5p target genes. Next the predicted results were docked with Human Ageing Genomic Resources (http://genomics.senescence.info/). Results The expression of miR-16-5p in peripheral blood leukocytes of aging rhesus macaques tended to increase with age (P<0.001). The predicted target genes of miR-16-5p were 539, and 19 of these were overlapped with Human Ageing Genomic Resources. It was showed that the 10 mRNA of 19 genes were detected and their expression were differences in the aging process of rhesus monkeys. The expressions of IKBKB, PIK3R1 and IRS1 were decreased gradually with aging compared with the infant group. Conclusions The miR-16-5p can be used as a biomarker of peripheral blood leukocytes in rhesus monkeys. And IKBKB, PIK3R1, IRS1 may be the target genes of miR-16-5p in the aging process of rhesus monkeys.

Dynamic expression of YTHDC1 in mice cerebellar development and human medulloblastoma

2019, 39(7):  1001-1006. 

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Objective To study the expression pattern of RNA m6A binding protein YTHDC1 during mouse cerebellar development and in human medulloblastoma. Methods Relative expression level of YTHDC1 RNA in the mice cerebellum, medulloblastoma and Daoy cell lines were analyzed based on RNA-seq results. Western blot and immunohistochemical staining were performed to detect the expression of YTHDC1 protein. The methyltransferase METTL3 was knocked down in Daoy cells, followed by m6A-IP-qPCR method to detect the change in YTHDC1 methylation level and the subsequent effect on the YTHDC1 protein level. Results During mice cerebellar development, the YTHDC1 expression remained constant in RNA level but showed a gradual reduction in protein level. Similarly, the expression of YTHDC1 protein in internal granular layer decreased significantly from postnatal day 7 (P7) to P60. Comparison between medulloblastoma tissues and adjacent para-carcinoma tissues, showed no apparent change of YTHDC1 in RNA level, but a significant decrease in protein level. Notably, YTHDC1 RNA methylation level increased in medulloblastoma, and the decreased m6A methylation level of YTHDC1 caused by METTL3 knockdown resulted in increased YTHDC1 protein expression. Conclusions In both cerebellar development and medulloblastoma, a dynamic change of YTHDC1 protein expression， but not YTHDC1 RNA, was observed. The RNA m6A modification of YTHDC1 can regulate its protein expression at post-transcriptional level, suggesting that RNA m6A methylation of YTHDC1 may impact on cerebellar development and medulloblastoma through regulating its protein expression.

Effects of the content of cordycepin, polysaccharides and water extract of cultivated and wild cordyceps militaris on anti-esophageal cancer cells activity

2019, 39(7):  1007-1012. 

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Objectiv By comparison of the content of cordycepin, polysaccharides and he anti-esophageal cancer cells activity of water extract of cultivated Cordyceps militaris and wild Cordyceps militaris, a preliminary study on the quality identification of Cordyceps militaris was carried out to provide certain reference value for the establishment of the quality evaluation system of Cordyceps militaris. Methods Using HPLC technology to detect the content of cordycepin in Cultivated Cordyceps militaris and Wild Cordyceps militaris. HPLC conditions—column: Zorbax SB-C18, 5μm, 250 mm×4.6 mm, Agi-lent, USA; the mobile phases: methanol (M,15% ) and water (W,85% ); flowing rate: 1.0 ml/min;temperature: 25℃; detection wave length: 260 nm; injection: 1μL. Phenol-sulfuric acid method was used to detect the content of polysaccharides. By MTS assay we detected the condition of cell proliferation treated with different concentrations of WECM. Annexin V/PI flow cytometry was used to determine the apoptosis of EC109 cells. Result HPLC results showed that the content of cordycepin in cultivated Cordyceps militaris and wild Cordyceps militaris ware 400.34 μg/g and 28.63 μg/g; the content of polysaccharides in cultivated Cordyceps militaris and wild Cordyceps militaris ware 4.256% and 3.115%，assaying by optimized phenol-sulfuric acid method; The results of MTS and flow cytometry indicated that the anticancer activity of the cultivated Cordyceps militaris was obviously more than wild Cordyceps militaris. Conclusion cultivated Cordyceps militaris is better than wild Cordyceps militaris in the the content of cordycepin, polysaccharides and the anticancer activity of water extract of Cordyceps militaris, it is indicated that Cultivated Cordyceps militaris is a good substitute for wild Cordyceps militaris.

Over-expression of spleen tyrosine kinase inhibits mouse osteoblasts apoptosis induced by oxidative stress

2019, 39(7):  1013-1018. 

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Objective To investigate the protective effect of spleen tyrosine kinase (SYK) on oxidative stress (OS)-induced injury and cell apoptosis in mouse osteoblasts MC3T3-E1 and the mechanism. Methods Osteoblast MC3T3-E1 was incubated with hydrogen peroxide (H2O2) to establish OS model. The mRNA expression of SYK was measured by quantitative real-time PCR (RT-qPCR). The protein expression levels of SYK, AKT, p-AKT and UCP2 were detected by Western blot. Recombinant plasmids pcDNA.3.1-SYK and pcDNA.3.1-UCP2 were constructed and separately transfected into MC3T3-E1. AKT phosphorylation was inhibited by AKT inhibitor HIMO. Cell apoptosis was measured by Annexin-V FITC/PImethod. Caspase-3 activity and reactive oxygen species (ROS) level were detected by kit methods. Results SYK was decreased by OS in osteoblasts (P<0.01). The overexpression of SYK significantly down-regulated cell apoptosis (P<0.05), caspase-3activity (P<0.01)，ROS level (P<0.01) induced by OS. Overexpression of SYK obviously increased AKT phosphorylation (P<0.05) and UCP2 expression (P<0.01). The inhibition of AKT phosphorylation could abolish the effects of SYK on cell apoptosis and UCP2 expression (P<0.01). UCP2 was overexpressed in MC3T3-E1 with SYK overexpression and AKT inhibition, and the cell apoptosis (P<0.05) and ROS level (P<0.01) were both markedly decreased. Conclusions The overexpression of SYK can prevent osteoblasts from injury induced by OS via regulating AKT/UCP2.

PM2.5 in utero exposure induces intrauterine growth retardation of mice through VEGF decrease caused by over-expression of HDAC2

2019, 39(7):  1019-1024. 

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Objective To investigate the role of histone acetylation in PM2.5 in utero exposure induced intrauterine growth retardation (IUGR). Method Pregnant mice were exposed to PM2.5 during the whole gestation, mice of control group were exposed to normal saline, non-treatment in blank control group; Birth weights were monitored; protein and mRNA levels of histone deacetylase (HDAC1/2/3 and vascular endothelial growth factor (VEGF) were measured by Western blot and Q-PCR methods; fluorometric assay was used to detect the activities of HDAC1/2/3; ChIP-PCR assay was employed to determine the binding levels of HDAC1 and HDAC2 with the promoter region of VEGF. Results Birth weights of mice in PM2.5 exposed group were significantly lower than those in control groups (p<0.05). HDAC1 and HDAC2 expression levels increased in trial groups, compared to controls (p<0.05). Protein and mRNA levels of VEGF decreased in PM2.5 exposed group, compared to those in control groups (p<0.05). Compared with control groups, HDAC2 activities went up after PM2.5 exposure (p<0.05). Binding levels of HDAC2 with promoter of VEGF increased (p<0.05). Conclusions PM2.5 exposure during gestation could induce IUGR, in our model, HDAC2 mediated a hypoacetylation near VEGF’s promoter, which caused a decrease of VEGF in placenta.

Over-expression of PTEN inhibits the protective effect of propofol against the apoptosis of H9c2 cardiacmyocytes induced by hydrogen peroxide

2019, 39(7):  1025-1030. 

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Objective To investigate the effect of phosphatase and tensin homolog deleted on chromosome 10(PTEN) on the apoptosis of H9C2 cardiomyocytes induced by H2O2. Methods H9C2 cardiomyocytes were divided into control group (normal cultured cells), H2O2 group (100 μmol/L H2O2 treatment for 24 h), propofol (5, 10, 30 μmol/L) group were treated with H2O2 for 24 h and then pretreated with propofol of different concentrations (5, 10, 30 μmol/L) for 1 h. The viability of cells was detected by MTT assay and the apoptosis rate was measured by flow cytometry. The expression of PTEN was detected by RT-qPCR; The viability, apoptosis, lactate dehydrogenase (LDH) , and reactive oxygen species (ROS) were analyzed of overexpression of PTEN. Results Compared with 0 μmol/L, 5 μmol/L and 10 μmol/L propofol significantly increased the activity of cardiomyocytes induced by H2O2, decreased the rate of apoptosis, and inhibited the expression of PTEN (P<0.05); 30 μmol/L propofol significantly inhibited cell viability and promoted apoptosis and the expression of PTEN (P<0.05); overexpression of PTEN could inhibit the protective effect of propofol against H2O2-induced H9C2 cells (P<0.05), and promote the expression of LDH and ROS (P<0.05). Conclusion PTEN inhibits the protective effect of propofol against H2O2-induced apoptosis of cardiomyocytes.

Toosendanin inhibits the growth of ovarian cancer and promotes apoptosis in nude mice

2019, 39(7):  1031-1035. 

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Objective To observe the inhibitory effect of toosendanin on tumor growth in nude mice bearing ovarian cancer and its mechanism. Methods Nude mice were randomLy divided into groups, and ovarian cancer cells were transplanted for modeling. Cyclophosphamide 0.4mL/d (10g/L) was injected intraperitoneally in positive group. In the experimental groups 1, 2 and 3, toosendanin 1, 10 and 20 mg/kg were dissolved in 0.02 mL/g normal saline for intraperitoneal injection once a day for 15 days. The tumor volume, mass and HE staining results were compared after treatment. Apoptotic indexes were detected by TUNEL method. The expressions of P53, Bax, Bcl-2 and caspase-9 were detected by RT-PCR. The expressions of P53, Bax, Bcl-2 and caspase-9 were detected by Western-blot method. Results Tumor volume and mass were the highest in the model group (P < 0.05), and apoptotic index was the lowest (P < 0.05). Tumor tissue in the model group had high cell density, disordered arrangement, hyperchromatic nuclei and obvious atypia. In experiment 1, 2, positive group and experiment 3 group, the cells were loosely arranged, the size of tumor cells was different, the nuclei were light stained and the nucleoli were small. Flake necrosis and apoptotic cells could be found in the tumors. The differences in the expression of Bax, Bcl-2 and caspase-9 and the expression of p-P53/P53 were statistically significant (P < 0.05). The differences between the experimental group and the positive group were similar, and the differences between the other two groups were statistically significant (P < 0.05). Conclusion Toosendanin can inhibit the growth of ovarian cancer in nude mice and promote the apoptosis of tumor cells.

Clinical features of adult patients with central nervous system Listeria monocytogenes infection

2019, 39(7):  1036-1039. 

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Objective To better understand the clinical characteristics of central nervous system Listeriosis. Methods The clinical data of adult patients with CNS Listeriosis admitted to PUMCH from 1999 to 2018 were collected. The clinical features were retrospectively analyzed. Results A total of 34 adult patients with CNS Listeriosis were enrolled in this study. The average age of all patients was (43±15) years, from which female patients took 68%. 94% patients had at least one underlying disease. All patients had fever, and 71% of patients appeared altered consciousness. Peripheral blood leukocytes and neutrophils were significantly increased while the level of lymphocytes was decreased; 85% of those patients had elevated C-reactive protein levels. 26% had poor clinical outcomes. Conclusions Adult patients with CNS Listeriosis are often associated with autoimmune diseases, and the clinical manifestation usually turned to be heavier. When patients have symptoms of central nervous system infection, we should consider Listeriosis, and empirical medication should cover Listeria.

Surgical strategy for temporal lobe epilepsy caused by cavernous malformation

2019, 39(7):  1040-1044. 

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Objective To investigate the surgical treatment principles of temporal lobe epilepsy caused by cavernous malformation and the surgical resection extent of the cavernous malformation. Methods The clinical data and follow-up results of 14 patients presented with temporal lobe epilepsy due to cavernous malformation admitted to the Department of Neurosurgery of Peking Union Medical College Hospital from May 2013 to Oct 2017 were retrospectively analyzed. The outcome of epilepsy control was assessed by the classification method proposed by International League Against Epilepsy in 2001. Results There were no deaths. No long-term neurological dysfunction was found in all patients. The last follow-up showed that 13 patients (92.8%) had no seizures throughout the year. Conclusion The patients with temporal lobe epilepsy due to cavernous malformation may receive maximized seizure control from surgical treatment, performed after the comprehensive evaluation of the patient's history, preoperative magnetic imaging and electrophysiological monitoring during the operation.

Analysis on the health medical examination of aged people at a community station in Beijing

2019, 39(7):  1045-1047. 

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Objective To analyse the health status of 268 aged people who underwent health checkups at their community stations. Methods To survey 268 aged people who came to the community station for a medical examination from 1 July 2018 to 30 September 2018 using group sampling. Results Among 268 aged people, the detection rate of arrhythmias was 48.89 %, overweight / obesity rate was 47.01 %, blood pressure increased detection rate was 41.79 %, blood lipids abnormal was 26,87 %, and fasting blood glucose increased 24.25 %. Conclusions In Sanli River district, There are several abnormal indexes are detected among the aged people, guidance should be given for those problems as arrhythmias, overweight / obesity, elevated blood pressure, abnormal blood lipids and elevated blood glucose.

Co-delivery of CRISPR plasmid and HDR donor can reduce the gene editing efficiency of CRISPR/Cas9 system

2019, 39(7):  1048-1054. 

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Objective To explore the effect of co-transfection of CRISPR plasmid and HDR donors on the gene editing efficiency of CRISPR/Cas9 system and a feasible solution to the problem. Methods T7E1 assay and RT-qPCR were used to compare the cleavage efficiency of Cas9 protein, Cas9 RNA and DNA level between cells transfected with CRSIPR plasmid only and cells co-transfected with CRISPR plasmid and HDR templates; RT-qPCR were used to detect the changes of intracellular Cas9 DNA and RNA level when reducing the quantity of co-transfected template DNA; RT-qPCR and T7E1 assay were used to measure the DNA level, RNA level and cleavage activity of Cas9 when delivering CRISPR plasmid followed by HDR donors. Results Compared with the group only CRISPR plasmid transfected, co-transfection of CRISPR plasmid and HDR donors significantly reduced the transfection efficiency of CRISPR plasmid (P<0.001) and the cleavage activity of Cas9 (P<0.001). But there was no significant difference in transfection efficiency of CRISPR plasmid and cutting efficiency of CRISPR/Cas9 system between only CRISPR plasmid transfected group and groups sequentially transfected with CRISPR plasmid and HDR donors. Conclusions Co-transfection of CRISPR plasmid and HDR donors can reduce the gene editing efficiency of CRISPR/Cas9 system and this problem can be solved by delivering CRISPR plasmid first and then HDR templates.

Neuronostatin inhibits contractile activity of uterine smooth muscle strips of rat

2019, 39(7):  1055-1056. 

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Research progress of ferroptosis in tumor

2019, 39(7):  1057-1060. 

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Ferroptosis, which is different from necrosis and apoptosis is a newly discovered programmed cell death. The characteristic of ferroptosis is the iron-dependent accumulation of lipid hydroperoxides to lethal levels. Ferroptosis is involved in the formation and development of different cancers such as hepatocellular cancer, pancreatic cancer, prostate cancer and breast cancer. SLC7A11 and GPX4 are the key regulators in ferroptosis; p53 and BECN1 participate in the ferroptosis via regulating SLC7A11 and GPX4; Inducing ferroptosis might become a new antitumor method in future.

Diabetes and pyroptosis

2019, 39(7):  1061-1065. 

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Pyroptosis is another form of cell death that is different from necrosis and apoptosis. For the body, cell coercion helps to maintain homeostasis, but excessive cell caliber can cause disease in the body，diabetes is one of them. Changes in metabolites and intestinal flora in pre-diabetes, diabetes can cause pyroptosis ，and pyroptosis further promotes the reduction of islet β-cell number and the formation of insulin resistance in insulin target organs, aggravating the development of diabetes and complications. Although the treatment based on pyroptosis is more and more concerned, it needs further study.

Research progress on the effects of feeder cells and its mechanism

2019, 39(7):  1066-1070. 

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Feeder cells have been widely used as an assistant method for the in vitro culture of mammal cells.It can simulate the condition of in vivo culture to a certain extent by the secretion of growth factors and the physical support. According to different culture purposes,we may choose specific feeder cells and dealing methods or modify feeder cells through gene editing to obtain the best culture results.

Exploration of incorporating formative assessment into clinical teaching assessment system of obstetrics and gynecology

2019, 39(7):  1071-1073. 

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Objective To explore the effect of formative assessment teaching in internship teaching of gynecology and obstetrics. Methods Since 2017, we tried mini-Clinical Evaluation Exercise (Mini-CEX) among obstetrics and gynecology interns. Results Compared with the traditional mode, students' ability of doctor-patient communication and surgical operation had been improved. Conclusions As the carrier of formative assessment, Mini - CEX should be incorporated into clinical teaching assessment system of obstetrics and gynecology.

Measures to improve the teaching ability of young teachers of aerospace medicine

2019, 39(7):  1074-1076. 

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Objective Young teachers are important factors for the sustainable development of aerospace medicine, so it is an important measure to enhance their teaching ability for the reform of curriculum system. Methods The training mechanism of young teachers has been set up with a series of measures, such as encouraging individualized teaching, promoting actual combat teaching and carrying out the teaching competitions. Results The quality of teachers has been enhanced gradually, and the reform of aerospace medical system has been advanced. Conclusion Through a series of measures, the teaching ability of young teachers has been significantly improved, and the satisfaction rate of students' classroom quality questionnaires has exceeded 95%.