Optimization of water extraction technology of Xiangqin jiere granules by orthogonal design based on G1-entropy weight compared with BP neural network / 中国药房

OBJECTIVE Optimizing the water extraction technology of Xiangqin jiere granules. METHODS The orthogonal test of 3 factors and 3 levels was designed， and comprehensive scoring was conducted for the above indexes by using G1-entropy weight to obtain the optimized water extraction technology of Xiangqin jiere granules with water addition ratio， extraction time and extraction times as factors， using the contents of forsythoside A， baicalin， phillyrin， oroxylin A-7-O-β-D-glycoside， wogonoside， baicalein and wogonin， and extraction rate as evaluation indexes. BP neural network modeling was used to optimize the network model and water extraction process using the results of 9 groups of orthogonal tests as test and training data， the water addition multiple， decocting time and extraction times as input nodes， and the comprehensive score as output nodes. Then the two analysis methods were compared by verification test to find the best water extraction process parameters. RESULTS The water extraction technology optimized by the orthogonal test was 8-fold water， extracting 3 times， extracting for 1 h each time. Comprehensive score was 96.84 （RSD＝0.90%）. The optimal water extraction technology obtained by BP neural network modeling included 12-fold water， extracting 4 times， extracting for 0.5 h each time. The comprehensive score was 92.72 （RSD＝0.77%）， which was slightly lower than that of the orthogonal test. CONCLUSIONS The water extraction technology of Xiangqin jiere granules is optimized successfully in the study， which includes adding 8-fold water， extracting 3 times， and extracting for 1 hour each time.

Kang-Ai Injection Inhibits Gastric Cancer Cells Proliferation through IL-6/STAT3 Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanisms underlying the proliferative inhibition of Chinese herbal medicine Kang-Ai injection (KAI) in gastric cancer cells.@*METHODS@#Gastric cancer cell lines MGC803 and BGC823 were treated by 0, 0.3%, 1%, 3% and 10% KAI for 24, 48 and 72 h, respectively. The cell proliferation was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The apoptosis and cell cycle were evaluated by flow cytometry. Interleukin (IL)-6 mRNA and protein expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immune sorbent assay (ELISA), respectively. The protein expression levels of cyclin A, cyclin E, cyclin B1, cyclin D1, p21, retinoblastoma (RB), protein kinase B (AKT), extracellular regulated protein kinases (ERK), signal transducer and activator of transcription (STAT) 1 and STAT3 were detected by Western blot.@*RESULTS@#KAI inhibited the proliferation of MGC803 and BGC823 gastric cancer cells in dose- and time-dependent manner. After treated with KAI for 48 h, the proportion of G1 phase was increased, expression level of cyclin D1 and phosphorylation-RB were down-regulated, whereas the expression of p21 was up-regulated (all P<0.01). Furthermore, 48-h treatment with KAI decreased the phosphorylation level of STAT3, inhibited the mRNA and protein expressions of IL-6 (all P<0.01). IL-6 at dose of 10 ng/mL significantly attenuated the proliferative effect of both 3% and 10% KAI, and recovered KAI-inhibited STAT3 phosphorylation and cyclin D1 expression level (all P<0.01).@*CONCLUSION@#KAI exerted an anti-proliferative function by inhibiting IL-6/STAT3 signaling pathway followed by the induction of G1 phase arrest in gastric cancer cells.

Correlation Between cyclin G1 Expression and Efficacy of Radiotherapy on Primary Hepatocellular Carcinoma / 肿瘤防治研究

Objective To investigate the correlation between cyclin G1 expression and the efficacy of radiotherapy on HCC. Methods The expression of cyclin G1 in biopsy specimens of 68 patients who received radiotherapy was detected by immunochemistry. The correlation between cyclin G1 expression and clinicopathological characteristics was analyzed by chi-square test. The correlation between cyclin G1 expression and OS or PFS was evaluated by Kaplan-Meier analysis. Univariate and multivariate analyses were used for the relation between clinicopathological characteristics and OS or PFS. Results The expression of cyclin G1 was related to portal vein tumor embolus, clinical stage and alpha fetoprotein. Survival analysis showed that the OS and PFS of patients with low expression of cyclin G1 were significantly higher than those with high cyclin G1 expression (P < 0.05). Multivariate analysis showed that cyclin G1 was an independent risk factor for DFS of patients with HCC. Conclusion High expression of cyclin G1 is an adverse prognostic factor for HCC patients who received radiotherapy.

DNA damage repair and cell cycle arrest / 国际生物医学工程杂志

In the face of DNA damage caused by various factors, cells have a set of response and repair mechanisms. Cell cycle arrest plays an important role in the DNA damage repair, which provides enough time for repairing damaged DNA. Research on cell cycle regulation focuses on cyclin-dependent protein kinases (CDKs) and cell cycle checkpoints. In the process of DNA damage repair, phosphatidylinositol-3-kinase like kinases (PIKKs) which are recruited to the DNA damage sites can activate cell cycle checkpoint-related proteins to halt cell cycle. In the common DNA damage repair pathways, such as base excision repair (BER), nucleotide excision repair (NER) , mismatch repair (MMR) , and DNA double-strand break repair, the recruitment of repair-related proteins also plays a role in the cell cycle regulation. In this paper, the relationship between the main forms of DNA damage repair and cell cycle arrest and relevant research progress were reviewed.

Effect of Wenxin Prescription on G1/S Cell Cycle Transformation in Atherosclerosis Rats / 中国实验方剂学杂志

Objective:To observe the effects of Wenxin prescription on the key targets of gap 1/synthesis （G₁/S） cell cycle transformation in rats with atherosclerosis （AS）， and reveal the mechanism of Wenxin prescription in the treatment of AS. Method:Ninety SPF Wistar rats were randomly divided into a normal group （<italic>n</italic>=6） and a modeling group （<italic>n</italic>=84）. The rats in the modeling group were fed on a high-fat diet （4% cholesterol， 0.5% sodium cholate， 0.2% propyl thiouracil， 10% lard， 5% sugar， and 80.3% basal feed） for 60 days， and intraperitoneally injected with 400 000 U·kg^-1vitamin D₃， once a week for three weeks. The model rats were then randomly divided into a model group， high-dose （24 g·kg^-1）， medium-dose （12 g·kg^-1）， and low-dose （6 g·kg^-1） Wenxin prescription groups， and a rosuvastatin （1.8 mg·kg^-1） group. The groups with drug intervention were treated correspondingly by gavage for 30 days. The rats in the model group were administered with an equal volume of distilled water. The general condition of rats was observed after treatment. The levels of high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and total cholesterol （CHO） were detected by enzyme-linked immunosorbent assay （ELISA）， and the atherosclerosis index （AI） was calculated. The pathological morphology of the coronary artery and aorta was observed by hematoxylin-eosin （HE） staining. The protein and mRNA expression of E2F transcription factor 1 （E2F1）， phosphorylated retinoblastoma protein （p-Rb）， cell division cycle 25 （Cdc25）， CyclinE， and CyclinD₁ was detected by Western blot and real-time fluorescence-based quantitative polymerase chain reaction （Real-time-PCR）， respectively. Result:Compared with the normal group， the model group showed intima thickening， smooth muscle proliferation， and plaque formation in the coronary artery and aorta， decreased HDL-C （<italic>P</italic><0.01）， increased LDL-C， CHO， and AI （<italic>P</italic><0.01）， elevated protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE and CyclinD₁ （<italic>P</italic><0.05）. Compared with the model group， the rosuvastatin group and the Wenxin prescription groups showed slight intimal hyperplasia and lumen narrowing of the coronary artery and aorta， decreased levels of LDL-C， CHO， and AI （<italic>P</italic><0.01）， and declining protein and mRNA expression of E2F1， Cdc25， p-Rb， CyclinE， and CyclinD₁ to varying degrees （<italic>P</italic><0.05）. Conclusion:Wenxin prescription can significantly inhibit the expression of key proteins and genes of the G₁/S cell cycle， regulate G₁/S cell cycle transformation， and reduce vascular smooth muscle and intimal hyperplasia in AS rats.

Arsenic on cholesterol efflux and the expression of ABCA1, ABCG1 and SRBI / 预防医学

Objective@#To explore the impact of arsenic on cholesterol efflux and the expression of ATP-binding cassette, sub-family A, member 1 ( ABCA1 ), ATP-binding cassette transporter G1 ( ABCG1 ), and scavenger receptor class B member I ( SRBI ) in macrophages, so as to provide the evidence for the mechanism of arsenic induced atherosclerosis.@*Methods@#The human myeloid leukemia mononuclear cells ( THP-1 ), induced by phorbol myristate acetate, and mouse primary macrophages were treated with 0, 0.625, 1.25, 2.5 and 5 μmol/L NaAsO2 for 48 hours. Then the cells treated with 2.5 μmol/L NaAsO2 were changed to arsenic free mediums for 48 hours and collected every 12 hours to analyze the time effect of arsenic. The expression levels of ABCA1, ABCG1 and SRBI were determined by quantitative polymerase chain reaction and western blotting. Cholesterol efflux rates were measured by 3H isotope tracer. @*Results@#Arsenic significantly down-regulated the expression levels of ABCA1 and ABCG1, and cholesterol efflux in a dose-dependent manner. The levels of ABCA1 mRNA decreased by 69% and 72%, the levels of ABCG1 mRNA decreased by 42% and 34%, and the rate of cholesterol efflux decreased by 55% and 59% in THP-1 and mouse primary macrophages cells treated with 5 μmol/L NaAsO2 ( all P<0.05 ). Arsenic had no significant effect on SRBI expression ( all P>0.05 ). Arsenic inhibited ABCA1 expression and cholesterol efflux in THP-1 in a time-dependent manner. Compared with cells before the exposure of arsenic, the level of ABCA1 mRNA and the rate of cholesterol efflux in THP-1 bottomed at 48 hours by 43% and 42%, and gradually recovered when arsenic was removed. @*Conclusions@#Arsenic inhibits cholesterol efflux by down-regulating the expression of ABCA1 and ABCG1 in macrophages.

O "menor infrator" na mídia: etnografia da criminalização da pobreza no G1 / The "juvenile offender" on the media: ethnography of poverty criminalization in G1 / El "menor infractor" en los medios: etnografía de la criminalización de la pobreza en el Portal G1

Resumo A mídia ocupa espaço expressivo na contemporaneidade, produzindo significações sobre fenômenos cotidianos, divulgando-os massivamente e, assim, definindo não só o que existe e o que faz parte da realidade, mas promovendo conotações valorativas sobre os acontecimentos. No contexto brasileiro, a mídia tem sido o principal vetor no que se refere às subjetivações sobre fenômenos da violência e da criminalidade. Tendo como objeto de pesquisa a repercussão no portal de notícias G1 de dois casos cujos protagonistas são jovens envolvidos em atos infracionais, este estudo mapeou as reportagens vinculadas a estes de modo a compreender as subjetividades produzidas, promovendo uma reflexão crítica a respeito da produção do "menor infrator" na mídia e seus desdobramentos. O método empregado para a realização do estudo foi a etnografia on-line, em consonância com os princípios da Teoria Ator-Rede. O rastreamento das matérias promoveu reflexões sobre a produção do "menor infrator", entre elas a percepção de que, tal como as políticas de subjetivação vetorizadas por esses dispositivos, as práticas que criminalizam a juventude pobre ocorrem de modo bastante sutil. É na constante ênfase dada pela mídia a elementos morais específicos e na arbitrariedade com a qual se destaca ou não a presença de transtornos psicológicos que vemos um caso ser tratado como punição merecida e o outro como tragédia. Assim, a veiculação de notícias que envolvem jovens tem se configurado uma importante tática de segregação e exclusão social, promovendo o apoio social às práticas de segurança pública que punem e exterminam jovens pobres.

Abstract The media produce meanings about everyday phenomena, spreading them massively and creating value connotations about events. In Brazil, this has been the main vector when it comes to the subjectivities about the phenomena of violence and criminality. Taking as research object the repercussion of the G1 news portal about two cases where the protagonists are young boys involved in infractions, this study mapped the reports related to them in order to understand the produced subjectivities, promoting a critical reflection on the production of the "juvenile offender" in the media and its consequences. The method used to carry out the study was an on-line ethnography, in accordance with the principles of the Actor-Network Theory. The tracking of the subjects allowed to reflect on the production of the "juvenile offender", being one of them the perception that, just like the policies of subjectivation vectored by these devices, the practices that criminalize the poor and young population happen in a very subtle way. The constant emphasis given by the media to specific moral elements and the arbitrariness with which the presence of psychological disorders is highlighted or not shows one case being treated as a deserved punishment and the other as a tragedy. This is how the news coverage involving young people serves as an important tactic of segregation and social exclusion, promoting social support for public safety and security practices that punish and exterminate poor young people.

Resumen Los medios ocupan un espacio significativo en la contemporaneidad al producir significaciones acerca de los fenómenos cotidianos, divulgarlos masivamente y definir no solo lo que existe y lo que es parte de la realidad, sino también promocionar connotaciones valorativas sobre los acontecimientos. En el contexto brasileño, los medios han sido el principal vector en lo que se refiere a las subjetivaciones sobre los fenómenos de la violencia y de la criminalidad. Teniendo como el objeto de la investigación la repercusión en el portal de noticias G1 de dos casos que tienen como protagonistas jóvenes que se involucraron en actos delictivos, la investigación ha mapeado los reportajes vinculados para comprender las subjetividades producidas al hacer una reflexión crítica a respeto de la producción acerca del "menor infractor" en los medios y sus implicaciones. Se utilizó la etnografía en línea según los principios de la teoría actor-red como método de investigación. El rastreo de los reportajes permitió reflexionar sobre la producción acerca del "menor infractor", entre ellas la percepción de que, tal como las políticas de subjetivación vectorizadas por estas plataformas, las prácticas que criminalizan la juventud pobre ocurren de modo bastante sutil. Sin embargo, es en el constante énfasis dado por los medios a elementos morales específicos y en la arbitrariedad con la cual se destaca o no la presencia de trastornos psicológicos que vemos un caso ser tratado con la debida punición y el otro, como una tragedia. La difusión de noticias que involucran a los jóvenes se traduce en una importante táctica de segregación y exclusión social, fomentando el apoyo social a prácticas de seguridad pública que castigan y exterminan a los jóvenes pobres.

Establishment of human embryonic stem cell line with forkhead box G1 gene knockout by CRISPR/Cas9 / 第二军医大学学报

Objective To construct human embryonic stem cell (hESC) line with forkhead box G1 (FOXG1) gene knockout by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing technology, and to investigate the role of FOXG1 gene in the early neural induction of hESCs. Methods Two guide RNAs (gRNAs) were transfected to induce FOXG1 gene large fragment knockout in hESCs by CRISPR/Cas9 gene editing technology. FOXG1 gene knockout hESCs were confirmed by monoclonal screening, sequencing and Western blotting analysis. The expression of the key markers including paired box 6 (PAX6), sex-determining region Y-box 2 (SOX2) and orthodenticle homeobox 2 (OTX2) was detected by immunofluorescence staining and qRT-PCR in the early process of neural induction before and after FOXG1 gene knockout. Results hESCs with FOXG1 gene large fragment knockout were successfully obtained by CRISPR/Cas9 gene editing technology. The results of immunofluorescence staining and qRTPCR suggested that FOXG1 deletion did not significantly influence the expression of PAX6, SOX2 and OTX2 during neural induction. Conclusion FOXG1 gene large fragment knockout in hESCs can be quickly induced by a pair of gRNAs cotransfection. FOXG1 deletion has no significant impacts on neural induction of hESCs.

New quinoline and anhydride compounds from fruits of Ficus carica / 中草药

Objective To investigate the chemical constituents of the fruits of Ficus carica. Methods The chemical constituents were isolated and purified with silica gel chromatography, Sephdex LH-20 and HPLC, and their structures were elucidated by spectral analysis including MS, NMR, IR spectroscopic analysis, elment analyzer, and chemical evidence. Results One new quinoline and two new anhydride compounds were isolated from ethyl acetate extract of the fruits of F. carica. The structures of the three new compounds were respectively defined as 4-hydroxyl-6-formyl-8α-methoxyl-quinoline-2-one (1), 5’β,6’α-[(di-ethyl)-5β,6β-cyclohexyl]- epoxyhexyl, cyclo-pentanhydride-[2,2,1]-2α,3β-cycloheptane (2), and 5’β,6’β-[di-(11-methylbutyl,11’-methylbutyl)-9,9’]-cyclo-8’- hexadiene-5α,6α-epoxyhexyl, cyclo-pentanhydride-[2,2,2,1]-2β,3α-cycloheptane (3). Conclusion These compounds were named figine K-1, figine G-1, and figine G-2, respectively.

Optimization of spray drying technology of Liqi Huoxue Compound extract by Box-Behnken response surface method combined with G1-entropy weight method / 中草药

Objective: To optimize the spray drying process of Liqi Huoxue Compound (LHC) extract and evaluate the quality of extract powder. Methods: The drug concentration, inlet air temperature and air flow were used as independent variables, and the comprehensive scores of ferulic acid, tanshinone IIA, salvianolic acid B, puerarin content and flour extraction rate were used as the response values, and the test data were binomial fitting. The mathematical relationship between the comprehensive scoring index and their respective variables was established, and Box-Behnken design-response surface method combined with G1-entropy weight method was used to optimize the spray drying process of LHC extract. Results: The optimal spray drying process was a specific solution gravity of 1.08, inlet air temperature of 140 ℃, and an air flow of 40 m3/h. Under such conditions, the average content of ferulic acid, tanshinone IIA, salvianolic acid B and puerarin by the spray drying was 0.300 4 mg/g, 0.338 0 mg/g, 18.526 0 mg/g and 7.508 8 mg/g, respectively. The flour yield was 81.49%, the overall score was 94.69, and the RSD value was 0.99%. Conclusion: The preferred spray drying process of LHC extract is stable and feasible.

Optimization of extraction technology and inclusion process of volatile oil for Qingxiangrukang Granules based on Box-Behnken and PCA-G1-entropy method and orthogonal design / 中草药

Objective: To optimize the extraction technology and inclusion process of volatile oil for Qingxiangrukang Granules (QG). Methods: Box-Behnken was used to optimize the extraction technology of volatile oil in QG. With yield of inclusion compound and inclusion rate of volatile oil-β-CD as evaluation indexes, and with volatile oil-pure water ratio, β-CD-volatile oil ratio, and inclusion time as investigate factors, the optimal inclusion technology for volatile oil of QG was ensured based on PCA-G1-entropy method and orthogonal design and by using colloid milling. Results: The optimum extraction technology of volatile oil were as follows: extracting time was 6 h, liquid-material ratio was 10, immersion was 0 h. The optimal inclusion technology for volatile oil were as follows: pure water-volatile oil ratio was 1:80, β-CD-volatile oil ratio was 6 g, and the inclusion time was 30 min. Under such condition, there was no significant difference between verification groups of three batches. Conclusion: This optimal extraction technology and inclusion process is stable and workable and can provide experimental basis for industrial production of QG.

Optimization of water extraction technology of Yiqi Huoxue Prescription based on orthogonal test design and BP neural network / 中草药

Objective: To optimize the water extraction technology parameters of Yiqi Huoxue Prescription (YHP). Methods: On the basis of single factor experiment, orthogonal experiment design was used to evaluate the transfer rate and extraction yield of salvianolic acid B and hydroxysafflower yellow A by using adding water, extraction time and soaking time as factors. The comprehensive score was obtained by G1-entropy weight method. The optimal water extraction technology was obtained by orthogonal test design, and another method-BP neural network modeling was used to optimize the network model and target optimization. The two analytical methods were compared in the verification experiment to find the optimal water extraction technology parameters of YHP. Results: Based on the comparison of the two analytical methods, it was found that the comprehensive score of the optimal water extraction technology obtained by orthogonal test analysis was slightly higher than that obtained by BP neural network modeling. Therefore, it was finally determined that the optimal water extraction technology parameters of YHP were as follow: water extraction for three times, soaking for 0.5 h, adding water of 20 times, and extracting time for 3.5 h. Conclusion: The optimal water extraction technology of YHP is stable and feasible, which provides new ideas and references for the development and modernization of new drugs of compound Chinese medicine.

Prokaryotic expression and hypoglycemic activity determination of insulin G1 from Conus geographus / 生物工程学报

Rapid reduction of postprandial blood glucose is very beneficial to diabetics. In order to shorten the onset time of recombinant insulin, the cone snail insulin G1 (cI G1) of Conus geographus was studied. First, the nucleotide sequence of recombinant cone snail proinsulin G1 (cPI G1) was designed and synthesized according to the genes of human proinsulin (hPI) and cPI G1. The codon was optimized according to Escherichia coli (E. coli) codon usage frequency. Then, the plasmid pET22b(+)-cPI G1 was constructed and the recombinant cPI G1 was expressed in E. coli BL21(DE3) host strain. The recombinant cPI G1 was then purified and cleaved specially by trypsin to generate the recombinant cI G1, and its potency is 25.9 IU/mg. Fasting blood glucose test (FBGT) and oral glucose tolerance test (OGTT) suggested that the recombinant cI G1 could rapidly reduce blood glucose in normal and streptozotocin (STZ)-induced diabetic mice, but only for a short duration. This study provides a technical reference for the development of recombinant fast-acting insulin.

Correlation analysis of single nucleotide polymorphism of ABCG1 gene and cerebral infarction in Li nationality of Hainan province / 中华行为医学与脑科学杂志

Objective To investigate the correlation between three single nucleotide polymorphisms of ABCG1 gene ( rs4148102,rs225445 and rs183436) and cerebral infarction of Li nationality in Hainan. Methods Totally 236 cases ( case group) of patients with cerebral infarction and 144 healthy controls (control group) of Li nationality who were treated in a hospital in Hainan province from January 2015 to June 2018 were selected. Genomic DNA from peripheral blood of patients in the two groups was extracted and gen-otype analysis was conducted using flight mass spectrometry. Snapstatus analysis software was used to analyze the association between haplotype and cerebral infarction,and Hardy-Weinberg equilibrium verification was performed. SPSS25. 0 was used to analyze the difference in genotype distribution and allele distribution. Re-sults The distribution frequencies of GG, AG, AA genotypes of rs4148102 in the case group ( 62. 7%, 25. 9%,11. 4%) and the control group(521. %,38. 2%,9. 7%) were significantly different (P<0. 05),and the occurrence frequencies of GG in the control group were significantly lower than those in the case group. The occurrence frequencies of TT,CT and CC of rs225445 were also statistically different between the two groups(36. 9%,41. 1%,22. 0% in case group,48. 6%,38. 2%,13. 2% in control group),and the occur-rence frequencies of CC in the control group was significantly lower than that in the case group. There were statistically significant differences in the frequencies of allele T and C of rs225445 and allele C and A of rs183436 between the two groups (P<0. 05). Conclusion Genotype GG of rs4148102 and genotype CC of rs225445 may be the susceptible factors of cerebral infarction in Li nationality in Hainan province.

Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment / 中国病理生理杂志

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model.Oil red O staining was used to determine whether the model was established successfully.miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h.The intracellular lipid droplets were observed by Oil red O staining.The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot.The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05).The intracellular cholesterol content was increased gradually (P<0.05) , and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group.Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05).No difference of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.

Estrogen and progesterone promote breast cancer cell proliferation by inducing cyclin G1 expression

Breast cancer is the most common cause of cancer among women in most countries (WHO). Ovarian hormone disorder is thought to be associated with breast tumorigenesis. The present study investigated the effects of estrogen and progesterone administration on cell proliferation and underlying mechanisms in breast cancer MCF-7 cells. It was found that a single administration of estradiol (E2) or progesterone increased MCF-7 cell viability in a dose-dependent manner and promoted cell cycle progression by increasing the percentage of cells in the G2/M phase. A combination of E2 and progesterone led to a stronger effect than single treatment. Moreover, cyclin G1 was up-regulated by E2 and/or progesterone in MCF-7 cells. After knockdown of cyclin G1 in MCF-7 cells using a specific shRNA, estradiol- and progesterone-mediated cell viability and clonogenic ability were significantly limited. Additionally, estradiol- and progesterone-promoted cell accumulation in the G2/M phase was reversed after knockdown of cyclin G1. These data indicated that estrogen and progesterone promoted breast cancer cell proliferation by inducing the expression of cyclin G1. Our data indicated that novel therapeutics against cyclin G1 are promising for the treatment of estrogen- and progesterone-mediated breast cancer progression.

Experimental study on the pancreatic cancer G1/S phase arrest byβ2-adrenergic antagonist / 西安交通大学学报(医学版)

Objective To investigate the effects of β2-adrenergic antagonist ICI118 ,551 on pancreatic cancer cell G1/S phase arrest and its action mechanism .Methods The cell cycle indexes were determined by the flow cytometry assay ;the expressions of Cyclin D1 and Cyclin E were analyzed by Western blot ;the activation of NF-κB was measured by electrophoretic mobility shift assay ;the proliferation of PanCa cells was determined by BALB/c athymic nude mice subrenal capsular assay .Results β2-adrenergic antagonist ICI118 ,551 significantly induced G1/S phase arrest compared with β1-adrenergic antagonist metoprolol in MIA PaCa-2 and BxPC-3 cell lines .ICI118 ,551 inhibited the expressions of Cyclin D1 and Cyclin E and reduced the activation of NF-κB .The proliferation of PanCa cells was strongly suppressed in the renal capsule xenografts in mice after ICI 118 ,551 treatment .Conclusion The blockage ofβ2-adrenoceptor markedly induces PanCa cells to arrest at G1/S phase and inhibits the proliferation of pancreatic cancer cells .

Optimization of extraction and purification technology for Huangqi Baihe Granules based on G1-entropy method and orthogonal design / 中草药

Objective To determine the optimum extraction and purification technology of Huangqi Baihe Granules (HBG) by using orthogonal design and combination empowerment based on G1-entropy method, so as to provide a reference for the industrial production. Methods With the ethanol extraction amount and paste-forming rate of calycosin7-O-β-D-glucopyranoside, hesperidin, crude polysaccharide as evaluation index, and with the extraction times, extraction time and the water adding amount as investigate factors, then the combination empowerment method based on G1-entropy method and orthogonal design were used to optimize the extraction technology of HBG. The same combined methods were used to optimize the purification technology of HBG with retention rate and removal rate of impurity of ethanol extract of calycosin7-O-β-D-glucopyranoside, hesperidin and crude polysaccharide, as evaluation index, and with the membrane pore diameter, operating pressure and filtration temperature of multi-channel tubular ceramic ultrafiltration membrane as investigate factors. Results The test results showed that the optimum extraction technology was as follows: Extracted 2 times with 20 times the amount of water and each for 75 min. The optimum ultrafiltration technology was as follows: Multi-channel tubular inorganic ceramic membrane at 50 nm, operating pressure at 0.10 MPa, filtration temperature at 45 ℃. Under such condition, there was no significant difference between verification groups of three batches. Conclusion The optimized extraction and purification process is stable and feasible by verification, which can provide experimental basis for industrial production of HBG.

Study on vacuum drying process of extract paste of Huangqi Baihe Granules based on Box-Behnken design-response surface and G1-entropy method / 中草药

Objective To optimize the vacuum drying process of extract paste of Huangqi Baihe Granules (HBG) and evaluate the physical quality of powder. Methods With drying temperature, vacuum degree, and material thickness as independent variables, the comprehensive evaluating indexes of content of calycosin 7-O-β-D-glucopyranoside, hesperidin, crude polysaccharide, ethanol extraction amount, and drying rate as response values, Box-Behnken design-response surface methodology and G1-entropy method were used to optimize the vacuum drying process. The similarity of fingerprints between extract powder dried by the optimized technology and extract paste was compared. Additionally, the properties of powder were evaluated comprehensively with nine physical indicators, including relative homogeneity index, bulk density, tap density, interparticle porosity, compressibility, Hausner ratio, angle of repose, moisture content, and hygroscopicity. The physical fingerprint of powder were established to evaluate the quality consistency of different batches of extract powder. Results The optimal drying parameter was as follows: the drying temperature was 68 ℃, the vacuum degree was 0.07 MPa, the material thickness was 6 mm. Three verification experiments were carried out under these conditions and the average comprehensive evaluating indexes of vacuum drying was 91.05, which was close to the model prediction 91.87, and the relative error was 0.89%. Compared with the extract paste, the similarity of fingerprint of extract powder were more than 0.91. The similarity of chemical and physical fingerprint of three batches of extract powder were higher than 0.99. Conclusion The optimized vacuum drying technology of extract paste of HBG is stable and feasible.

Interaction between hepatitis B virus X protein and mitochondrial elongation factor G1 / 中华实验和临床病毒学杂志

Objective@#To study the interaction between hepatitis B virus X protein(HBx) and mitochondrial elongation factor G1 (EFG1) in yeast cells or hepatoma cells.@*Methods@#After verification the interaction between HBx and EFG1 by CytoTrap yeast two-hybrid system, EFG1 genome was amplified by means of polymerase chain reaction(PCR) and cloned into the pcDNA3.1/myc-His(-)A vector, following verification by sequencing. Expression of HBx and EFG1 protein was verified in Huh7 cells. Then the recombinant vector pcDNA3.1/myc-His(-)A-EFG1 and pFLAG-CMV-2-HBx were transfected into Huh7 cells; 48 h later, the cells were lysed. The direct interaction between HBx and EFG1 was further confirmed by the Co-immunoprecipitation (Co-IP) assay.@*Results@#The interaction between HBx and EFG1 was successfully verified by CytoTrap yeast two-hybrid system. The recombined plasmid pcDNA3.1/myc-His(-)A-EFG1 was obtained. Furthermore, Co-IP assay was used to confirm the interaction between HBx and EFG1 in Huh7 cells.@*Conclusions@#The direct interaction between HBx and EFG1 was confirmed. Therefore our findings provide experimental basis for the influence of HBx protein on the expression of mitochondrial protein and provide new insights into the pathogenesis of HBx in the development of hepatocellular carcinoma.