Intestinal and pharyngeal microbiota in early neonates: an analysis based on high-throughput sequencing / 中国当代儿科杂志

OBJECTIVES@#To investigate the distribution characteristics and correlation of intestinal and pharyngeal microbiota in early neonates.@*METHODS@#Full-term healthy neonates who were born in Shanghai Pudong New Area Maternal and Child Health Hospital from September 2021 to January 2022 and were given mixed feeding were enrolled. The 16S rRNA sequencing technique was used to analyze the stool and pharyngeal swab samples collected on the day of birth and days 5-7 after birth, and the composition and function of intestinal and pharyngeal microbiota were analyzed and compared.@*RESULTS@#The diversity analysis showed that the diversity of pharyngeal microbiota was higher than that of intestinal microbiota in early neonates, but the difference was not statistically significant (P>0.05). On the day of birth, the relative abundance of Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05). On days 5-7 after birth, the relative abundance of Actinobacteria and Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05), and the relative abundance of Firmicutes in the intestine was significantly lower than that in the pharynx (P<0.05). At the genus level, there was no significant difference in the composition of dominant bacteria between the intestine and the pharynx on the day of birth (P>0.05), while on days 5-7 after birth, there were significant differences in the symbiotic bacteria of Streptococcus, Staphylococcus, Rothia, Bifidobacterium, and Escherichia-Shigella between the intestine and the pharynx (P<0.05). The analysis based on the database of Clusters of Orthologous Groups of proteins showed that pharyngeal microbiota was more concentrated on chromatin structure and dynamics and cytoskeleton, while intestinal microbiota was more abundant in RNA processing and modification, energy production and conversion, amino acid transport and metabolism, carbohydrate transport and metabolism, coenzyme transport and metabolism, and others (P<0.05). The Kyoto Encyclopedia of Genes and Genomes analysis showed that compared with pharyngeal microbiota, intestinal microbiota was more predictive of cell motility, cellular processes and signal transduction, endocrine system, excretory system, immune system, metabolic diseases, nervous system, and transcription parameters (P<0.05).@*CONCLUSIONS@#The composition and diversity of intestinal and pharyngeal microbiota of neonates are not significantly different at birth. The microbiota of these two ecological niches begin to differentiate and gradually exhibit distinct functions over time.

Bioinformatics Analysis of Core Genes and Key Pathways in Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To screen differentially expressed gene (DEG) related to myelodysplastic syndrome (MDS) based on Gene Expression Omnibus (GEO) database, and explore the core genes and pathogenesis of MDS by analyzing the biological functions and related signaling pathways of DEG.@*METHODS@#The expression profiles of GSE4619, GSE19429, GSE58831 including MDS patients and normal controls were downloaded from GEO database. The gene expression analysis tool (GEO2R) of GEO database was used to screen DEG according to | log FC (fold change) |≥1 and P<0.01. David online database was used to annotate gene ontology function (GO). Metascape online database was used to enrich and analyze differential genes in Kyoto Encyclopedia of Genes and Genomes (KEGG). The protein-protein interaction network (PPI) was constructed by using STRING database. CytoHubba and Mcode plug-ins of Cytoscape were used to analyze the key gene clusters and hub genes. R language was used to diagnose hub genes and draw the ROC curve. GSEA enrichment analysis was performed on GSE19429 according to the expression of LEF1.@*RESULTS@#A total of 74 co-DEG were identified, including 14 up-regulated genes and 60 down regulated genes. GO enrichment analysis indicated that BP of down regulated genes was mainly enriched in the transcription and regulation of RNA polymerase II promoter, negative regulation of cell proliferation, and immune response. CC of down regulated genes was mainly enriched in the nucleus, transcription factor complexes, and adhesion spots. MF was mainly enriched in protein binding, DNA binding, and β-catenin binding. KEGG pathway was enriched in primary immunodeficiency, Hippo signaling pathway, cAMP signaling pathway, transcriptional mis-regulation in cancer and hematopoietic cell lineage. BP of up-regulated genes was mainly enriched in type I interferon signaling pathway and viral response. CC was mainly enriched in cytoplasm. MF was mainly enriched in RNA binding. Ten hub genes and three important gene clusters were screened by STRING database and Cytoscape software. The functions of the three key gene clusters were closely related to immune regulation. ROC analysis showed that the hub genes had a good diagnostic significance for MDS. GSEA analysis indicated that LEF1 may affect the normal function of hematopoietic stem cells by regulating inflammatory reaction, which further revealed the pathogenesis of MDS.@*CONCLUSION@#Bioinformatics can effectively screen the core genes and key signaling pathways of MDS, which provides a new strategy for the diagnosis and treatment of MDS.

Efficacy and tolerability of dolutegravir plus lamivudine as a switch simplified strategy in treatment-experienced human immunodeficiency virus/acquired immunodeficiency syndrome patients / 中华传染病杂志

Objective:To evaluate the efficacy and tolerability of the dual therapy of dolutegravir (DTG) plus lamivudine (3TC) as a switch simplified strategy in treatment-experienced human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients.Methods:Treatment-experienced HIV/AIDS patients who switched to a dual therapy containing DTG (50 mg, once daily) plus 3TC (300 mg, once daily) were included in Beijing You′an Hospital, Capital Medical University from September 2016 to May 2019. HIV RNA, CD4 + T lymphocyte count, blood lipid indexes, renal function indexes were collected when patients changed the treatment regimen (baseline) and after 48 weeks of treatment. Efficacy (HIV RNA<50 copies/mL) and safety of the dual therapy were analyzed. Statistical comparisons were performed using the Wilcoxon matched-pairs signed rank test. Results:The reasons for 33 patients switching the treatment regimen were virologic failure (four cases, 12.1%), simplification of regimen (11 cases, 33.3%), and drug toxicity (18 cases, 54.5%). The patients were treated with anti-retroviral therapy (ART) for 2.13 (1.05, 4.23) years before regimens switching. Twenty-nine (87.9%) patients were virologically suppressed at baseline, and four (12.1%) patients were virological failure. After switching to DTG plus 3TC, all 33 patients showed HIV RNA<50 copies/mL after 48 weeks of treatment. The baseline CD4 + T lymphocyte count was 543 (363, 595)/μL. After switching the treatment regimens for 48 weeks, CD4 + T lymphocyte count was significantly increased to 625 (455, 651)/μL, and the difference was statistically significant ( Z=3.14, P=0.002). Compared with baseline, low-density lipoprotein-cholesterol was increased after 48 weeks of treatment (2.35(1.80, 3.08) mmol/L vs 3.12(2.74, 3.87) mmol/L), while triglyceride (2.21(1.27, 4.37) mmol/L vs 1.61(1.20, 2.22) mmol/L), the ratio of total cholesterol to high-density lipoprotein-cholesterol (5.02 (4.13, 6.40) vs 4.70 (3.55, 5.35)) and estimated glomerular filtration rate (106.4(78.2, 118.2) mL/(min·1.73 m 2) vs 88.6 (75.7, 107.9) mL/(min·1.73 m 2)) were decreased. The differences were all statistically significant ( Z=4.89, 2.37, 2.09 and 2.83, respectively, all P<0.050). No patient discontinued due to adverse events. Conclusions:The use of dual therapy containing DTG and 3TC is effective and well-tolerated in treatment-experienced HIV/AIDS patients under any prior ART without significant adverse events.

CXCL8 Promotes M2 Macrophage Chemotaxis and Infiltration in Colorectal Cancer Microenvironment / 中国生物化学与分子生物学报

CXC chemokine ligand 8 (CXCL8) is highly expressed in many human tumors including colorectal cancer, and it can promote the malignant progression of tumors. It was reported that M2 macrophages were abundant in colorectal cancer microenvironment, but whether CXCL8 affects the infiltration of M2 macrophages and its potential mechanism are not yet clear. The study aimed to investigate the effect of CXCL8 on M2 macrophage infiltration and chemotaxis in the colorectal cancer. Firstly, we analyzed the CXCL8 expression and immune cell infiltration in human colorectal cancer tissues from TCGA RNA-seq data. The expression of CXCL8 was verified by immunohistochemistry in tissues obtained from Shanxi Provincial Cancer Hospital. Then, Western blot and qRT-PCR were employed to detect CXCL8 expression in five colorectal cancer cell lines. THP-1 cells were allowed to differentiate into M2 macrophages via the phorbol myristate acetate (PMA) and IL-4 treatment, followed by detection of the chemotaxis of M2 macrophages towards HCT116, SW480 and CXCL8-HCT116, CXCL8-SW480 cell lines. HCT116 and SW480 cells were treated with interleukin 1β (IL-1β) to detect the expression of CXCL8, and co-cultured with M2 macrophages to analyze the chemotactic activity. The results revealed that the expression of CXCL8 was increased in pairs of CRC tissues versus normal adjacent tissues, and there were more M2 macrophage infiltration in cancer tissues with high expression of CXCL8. The mRNA and protein expression of CXCL8 in HCT116 and SW480 were increased after the IL-1β treatment (P < 0. 05). We confirmed that CXCL8 is a chemotactic factor for M2 macrophages by transwell assays (P<0. 05). In conclusion, CXCL8 in colorectal cancer cells can be induced by IL-1β in colorectal cancer cells and the upregulation of CXCL8 can promote the chemotaxis of M2 macrophages. The massive infiltration of M2 macrophages in colorectal cancer microenvironment may be related with the increased expression of CXCL8.

Study on synergistic inhibitory effect of ruxolitinib and decitabine on growth of HEL cells with TET2 knockdown / 中国药理学通报

Aim To explore the cytotoxic and synergistic effects of decitabine and ruxolitinib on HEL cells with TET2 knockdown. Methods Stable TET2 knockdown by shRNA was established in HEL cell line. The change of cell proliferation was measured by CCK-8 assay. The median lethal dose (IC50) and colony formation assay were used to evaluate the cytotoxic effects of decitabine and ruxolitinib, the synergistic effects of which was further analyzed by Chou-Talalay method. Results The inhibition of TET2 increased the proliferative capacity of HEL cells. HEL cell lines became resistant to decitabine following shRNA-media- ted TET2 inactivation. Colony formation assay showed that the drug sensitivity of decitabine and ruxolitinib both decreased in TET2 knockdown HEL cells. The synergistic inhibitory effects of ruxolitinib and decitabine on TET2 knockdown HEL cells were observed. Conclusion The combination of ruxolitinib and decitabine may be an effective therapeutic strategy for accelerated or blast phase MPN patients with JAK2V6m and TET2 mutations.

Anterior Gradient 3 Promotes Breast Cancer Development and Chemotherapy Response / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#Anterior gradient 3 (AGR3) belongs to human anterior gradient (AGR) family. The function of AGR3 on cancer remains unknown. This research aimed to investigate if AGR3 had prognostic values in invasive ductal carcinoma (IDC) of breast cancer and could promote tumor progression. @*Materials and Methods@#AGR3 expression was detected in breast benign lesions, ductal carcinoma in situ and IDC by immunohistochemistry analysis. AGR3’s correlations with clinicopathological features and prognosis of IDC patients were analyzed. By cell function experiments, collagen gel droplet-embedded culture drug sensitivity test and cytotoxic analysis, AGR3’s impacts on proliferation, invasion ability, and chemotherapeutic drug sensitivity of breast cancer cells were also detected. @*Results@#AGR3 was up-regulated in luminal subtype of histological grade I-II of IDC patients and positively correlated with high risks of recurrence and distant metastasis. AGR3 high expression could lead to bone or liver metastasis and predict poor prognosis of luminal B. In cell lines, AGR3 could promote proliferation and invasion ability of breast cancer cells which were consistent with clinical analysis. Besides, AGR3 could indicate poor prognosis of breast cancer patients treated with taxane but a favorable prognosis with 5-fluoropyrimidines. And breast cancer cells with AGR3 high expression were resistant to taxane but sensitive to 5-fluoropyrimidines. @*Conclusion@#AGR3 might be a potential prognostic indicator in luminal B subtype of IDC patients of histological grade I-II. And patients with AGR3 high expression should be treated with chemotherapy regimens consisting of 5-fluoropyrimidines but no taxane.

Research status of polycythemia vera in China / 中国实用内科杂志

Polycythemia vera is a clonal malignant hematopoietic disorder that results from genetic alterations in hematopoietic stem cells, which is characterized by two or three-line blood cells increase, and mainly by erythrocytosis. This article reviews research situation of PV in China, including pathogenesis, clinical features, disease progression and therapeutic options, which helps clinical specialists to carry out precise diagnosis and treatment.

Impaired Hypothalamic Regulation of Sympathetic Outflow in Primary Hypertension / 神经科学通报·英文版

The hypothalamic paraventricular nucleus (PVN) is a crucial region involved in maintaining homeostasis through the regulation of cardiovascular, neuroendocrine, and other functions. The PVN provides a dominant source of excitatory drive to the sympathetic outflow through innervation of the brainstem and spinal cord in hypertension. We discuss current findings on the role of the PVN in the regulation of sympathetic output in both normotensive and hypertensive conditions. The PVN seems to play a major role in generating the elevated sympathetic vasomotor activity that is characteristic of multiple forms of hypertension, including primary hypertension in humans. Recent studies in the spontaneously hypertensive rat model have revealed an imbalance of inhibitory and excitatory synaptic inputs to PVN pre-sympathetic neurons as indicated by impaired inhibitory and enhanced excitatory synaptic inputs in hypertension. This imbalance of inhibitory and excitatory synaptic inputs in the PVN forms the basis for elevated sympathetic outflow in hypertension. In this review, we discuss the disruption of balance between glutamatergic and GABAergic inputs and the associated cellular and molecular alterations as mechanisms underlying the hyperactivity of PVN pre-sympathetic neurons in hypertension.

Rictor regulates mitochondrial calcium signaling in mouse embryo stem cell-derived cardiomyocytes / 浙江大学学报·医学版

OBJECTIVE@#To explore the expression, localization and regulatory effect on mitochondrial calcium signaling of Rictor in embryonic stem cell-derived cardiomyocytes (ESC-CMs).@*METHODS@#Classical embryonic stem cell cardiomyogenesis model was used for differentiation of mouse embryonic stem cells into cardiomyocytes. The location of Rictor in ESC-CMs was investigated by immunofluorescence and Western blot. The expression of Rictor in mouse embryonic stem cells was interfered with lentiviral technology, then the superposition of mitochondria and endoplasmic reticulum (ER) in ESC-CMs was detected with immunofluorescence method; the cellular ultrastructure of ESC-CMs was observed by transmission electron microscope; the mitochondrial calcium transients of ESC-CMs was detected by living cell workstation;immunoprecipitation was used to detect the interaction between 1,5,5-trisphosphate receptor (IP3 receptor, IP3R), glucose-regulated protein 75 (Grp75) and voltage-dependent anion channel 1 (VDAC1) in mitochondrial outer membrane; the expression of mitochondrial fusion protein (mitonusin-2, Mfn2) was detected by Western blot.@*RESULTS@#Rictor was mainly localized in the endoplasmic reticulum and mitochondrial-endoplasmic reticulum membrane (MAM) in ESC-CMs. Immunofluorescence results showed that Rictor was highly overlapped with ER and mitochondria in ESC-CMs. After mitochondrial and ER were labeled with Mito-Tracker Red and ER-Tracker Green, it was demonstrated that the mitochondria of the myocardial cells in the Rictor group were scattered, and the superimposition rate of mitochondria and ER was lower than that of the negative control group (<0.01). The MAM structures were decreased in ESC-CMs after knockdown of Rictor. The results of the living cell workstation showed that the amplitude of mitochondrial calcium transients by ATP stimulation in ESC-CMs was decreased after knockdown of Rictor (<0.01). The results of co-immunoprecipitation showed that the interaction between IP3R, Grp75 and VDAC1 in the MAM structure of the cardiomyocytes in the Rictor group was significantly attenuated (<0.01); the results of Western blot showed that the expression of Mfn2 protein was significantly decreased (<0.01).@*CONCLUSIONS@#Using lentiviral technology to interfere Rictor expression in mouse embryonic stem cells, the release of calcium from the endoplasmic reticulum to mitochondria in ESC-CMs decreases, which may be affected by reducing the interaction of IP3R, Grp75, VDAC1 and decreasing the expression of Mfn2, leading to the damage of MAM structure.

Research Progress of Folate Metabolic Gene Polymorphism / 医药导报

Through summarizing and analyzing a large number of documents and reports of large academic conferences in China, the current situation and the prospect of individualized drug delivery model were analyzed based on folate metabolic gene in pharmaceutical care. Folate metabolic genemethylenetetrahydrofolate reductase (MTHFR) C677T and methionine synthase reductase (MTRR) A66G were related with development of multiple diseases. Its polymorphism guidesindividualized drug administrationto increase the efficacy of drugs or decrease adverse effects.

Study on relationship between hemoglobin content and blood pressure in pregnant women in Zhoushan islands / 中华疾病控制杂志

Objective To investigate the relationship between hemoglobin and blood pressure of pregnant women in Zhoushan islands, so as to provide scientific evidence for the etiological study of gestational hypertension. Methods A retrospective study was conducted among 1 383 pregnant women who received perinatal care in Zhoushan Maternal and Child Health Hospital of Zhejiang Province from January 2017 to June 2018. Pregnant women were monitored for hemoglobin content and blood pressure in the early, middle and late pregnancy. The multivariate linear regression was used to analyze the relationship between hemoglobin content and blood pressure in different pregnancy. Results The incidence of anemia in early, middle and late pregnancy was 7.74%, 25.45% and 15.76% respectively. The multivariate linear regression showed that hemoglobin levels during pregnancy had effects on systolic blood pressure in early, middle and late pregnancy, and the earlier hemoglobin levels were monitored, the more obvious the effect on systolic blood pressure was.With the increase of hemoglobin level, systolic blood pressure increased, such as the effect of hemoglobin on systolic blood pressure in early pregnancy, mid-pregnancy and late pregnancy. Hemoglobin of first trimster had the greatest effect (β=0.10, P<0.001), Hemoglobin of second trimester had no obvious effect, and that of third trimester had the second effect (β=0.04, P=0.027).Hemoglobin levels and diastolic blood pressure levels were similar to their relationship with systolic blood pressure. Conclusions Hemoglobin levels during pregnancy have significant effects on systolic and diastolic blood pressure in first, second and third trimsters of pregnancy. Regular measurement of hemoglobin levels during pregnancy can improve the health of pregnant women.

Professor 's experience of acupuncture and medication for allergic rhinitis / 中国针灸

Professor - believes that allergic rhinitis is mainly caused by deficiency of lung-spleen-kidney, combined with the attack of external pathogens, especially wind pathogens, which invade nasal orifices. Thus, three principles are established: syndrome differentiation and treatment, internal and external coherence, strengthening the root to eliminate pathogenic factors. For clinical practice, professor proposes the original acupuncture which takes the conception vessel and governor vessel as the general principles, takes syndrome differentiation as core, and takes regulating and as the major method. This acupuncture method integrates spirit and , bases on verified method and acupoint, combines acupuncture and medication, and achieves satisfactory results.

Dosimetry study of fourtypes of radiotherapy plan optimization methods in the hypofractionated radiotherapy for lung cancer / 中华放射肿瘤学杂志

Objective To discuss the dosimetric differences in the planning methods between physical and biological optimization during thehypofractionated radiotherapy for lung cancer.MethodsTen cases of non-small cell lung cancer (NSCLC) receiving radiotherapy were selected in this retrospective study.The VMAT plans for all patients were re-designed by physical functions (DV group),biological combined with physical functions (DV+EUD group and EUD+DV group) and biological functions (EUD group).The constrained functions were different,whereas the constrained conditions and optimized parameters were identical among four groups.The dosimetric differences among four optimization methods during thehypofractionated radiotherapy for lung cancer were evaluated through calculating and analyzing each dosimetry parameter.Results For the target area,the equivalent uniform dose was approximate between the EUD and EUD+DV groups.The EUD in these two groups was approximately 2.8％-3.6％ and 3.2％-3.7％ higher than those in the DV and DV+EUD groups.The average tumor control probability (TCP) in the EUD and EUD +DV groupswas considerably higher than those in the other two groups (both P＜0.05).The homogeneity index (HI) significantly differed (all P＜0.05),whereas the conformity index (CI) did not differ (all P＞0.05).For the organ at risk (OAR) area,the differences of EUD,V5,V1o,V20,V30 of normal lung tissues and the difference of dosimetry parameters in heart and spinal cord were not statistically significant (all P＞0.05).The mean dose of all lungs in the EUD and EUD+DV groupswas slightly lower than those in the other two groups.ConclusionsBiological optimization method has certain advantages in increasing EUD and TCP in the target area and decreasing the irradiation dose of normal lung tissues,which provides references for selecting the optimization method with biological functions in clinical practice.

Effects of inhibitory concentration minocycline on the proliferation, differentiation, and mineralization of osteoblasts / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effect of the inhibitory concentration minocycline on the proliferation, differentiation, and expression of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteopontin (OPN) mRNA of osteoblasts.</p><p><b>METHODS</b>Primary osteoblasts were cultured in osteogenic induction medium containing 0, 0.1, 0.5, 1, 10 μg·mL⁻¹ minocycline. Cell counting kit-8 was used to observe cell proliferation. ALP activity assay, alizarin red S staining, and real-time quantitative polymerase chain reaction (PCR) were used to determine cell differentiation and mineralization.</p><p><b>RESULTS</b>The groups with 0.1, 0.5, 1 μg·mL⁻¹ minocycline promoted cell proliferation. The mRNA expression levels of ALP and Runx2 were up-regulated. Osteoblast-mediated mineralization was increased. The group with 1 μg·mL⁻¹ showed maximal promotion effect (P<0.05). When the concentration increased to 10 μg·mL⁻¹, the promoting effect began to decline, and the ALP activity and OPN expression were significantly inhibited (P<0.01).</p><p><b>CONCLUSIONS</b>Appropriate concentration of minocycline can promote osteoblasts proliferation, up-regulate the expression levels of Runx2, ALP and OPN, and increase the differentiation and mineralization of osteoblasts.</p>

Education via WeChat improves glucose control and pregnant outcomes in women with gestational diabetes mellitus / 中华围产医学杂志

Objective To evaluate the effect of strengthening nutrition intervention in gravidas with gestational diabetes mellitus (GDM) with WeChat on blood glucose control and pregnant outcomes.Methods A total of 410 gravidas,diagnosed with GDM and treated in the Department of Clinical Nutrition of Shanghai General Hospital from October 2015 to April 2016,were enrolled and randomly divided into two groups (n=205).The control group received traditional nutrition clinic education only,while the intervention group was given strengthened nutrition education through WeChat in addition to traditional education.Blood glucose level and insulin dosage were followed up after one,two and four weeks of intervention.Pregnant outcomes and patient satisfaction were investigated on 42 d after delivery.T test,Chi-square test and non-parametric test were used for statistical analysis.Results (1) Two weeks after the intervention,the average 1-hour postprandial blood glucose in the intervention group was lower than in the control group [(7.46± 1.01) vs (7.68± 1.06) mmol/L,t=2.243,P=0.025].After 4 weeks,both 1-and 2-hour postprandial blood glucose levels of the intervention group were lower than those of the control group [(7.03±0.65) vs (7.33±0.63) mmol/L,t=4.629,P＜0.05;(6.00±0.65) vs (6.21 ±0.62) mmol/L,t=3.153,P＜0.05] and more gravidas achieved euglycemia [79.9％ (151/189) vs 60.8％ (113/186),x2=16.483,P＜0.001].(2) Compared with the control group,the intervention group had a higher vaginal delivery rate [38.7％ (72/186) vs 50.5％ (95/188),x2=5.288,P=0.021] and a lower rate of postpartum complications [9.1％ (17/186) vs 2.1％ (4/188),x2=7.394,P=0.007].All of the gravidas in the intervention group were satisfied with the WeChat intervention except one lost to follow up [99.5％ (203/204)].Conclusions Strengthening nutrition education through WeChat is much more effective than traditional nutritional outpatient education alone in order to achieve a better control of blood glucose and improve pregnant outcomes in GDM women.This intervention is highly acceptable to gravidas and can be further extensively applied in nutrition clinic.

Effect of toosendanin on invasion and migration of human ovarian cancer cells / 中国病理生理杂志

AIM:To investigate the effect of toosendanin(TSN)on invasion and migration abilities of human ovarian cancer cells and the related mechanism.METHODS:The human ovarian cancer cell lines CAVO-3 and SKVO-3 were treated with TSN at different concentrations.The cell viabilty at 12,24,48,72 and 96 h after TSN treatment was measured by CCK-8 assay.Scratch wound healing assay and Transwell assay were employed to measure the invasion and migration abilities of CAVO-3 cells.The protein expression of nuclear factor-κB(NF-κB)p65,E-cadherin,N-cadherin,vimentin and Snail was determined by Western blot.RESULTS:TSN significantly inhibited the viability of CAVO-3 and SKVO-3 cells(P<0.05 ).Compared with control group ,the migration and invasion abilities of CAVO-3 cells in TSN group decreased significantly(P<0.05).In addition,the expression of NF-κB p65 and E-cadherin protein increased no-tably,followed with N-cadherin,vimentin and Snail protein decreased significantly(P<0.05).However,the inhibitor of NF-κB BAY11-7082 reversed the impact above.Compared with TSN group ,the migration and invasion abilities in TSN +BAY11-7082 group increased significantly(P<0.05).The protein expression of E-cadherin also decreased notably ,fol-lowed with the protein expression of N-cadherin,vimentin and Snail increased significantly(P<0.05).CONCLUSION:TSN inhibits the invasion and migration abilities of human ovarian cancer cells ,which is related to the inhibition of epitheli-al-mesenchymal transition process mediated by NF-κB/Snail signaling pathway.

Relationship between TGF-β3 and anti-proliferative effect of ursolic acid in human colon cancer cells / 中国药理学通报

Aim To investigate the role of TGF-β3 in the anti-proliferation effect of ursolic acid(UA)in co-lon cancer cells and the possible molecular mechanism underlying this effect.Methods We introduced crys-tal violet staining,flow cytometry and Western blot as-say to determine the effect of UA on proliferation and apoptosis in HCT1 1 6 cells.The levels of TGF-β3, Smad2 /3 and β-catenin in HCT1 1 6 cell were evaluated by RT-PCR and Western blot.Finally,TGF-β3 inhibi-tor and recombinant adenovirus,and luciferase reporter assay were used to analyze the possible mechanism through which TGF-β3 mediated the anti-cancer effect of UA in HCT1 1 6 cells.Results UA inhibited the proliferation and induced apoptosis apparently in HCT1 1 6 cells.UA down-regulated TGF-β3 both in mRNA and in protein level.Meanwhile,UA decreased the phosphorylation of Smad2 /3 concentration depend-ently,although no significant effect was found on the total protein level of Smad2 /3 in HCT1 1 6 cells.Over-expression of TGF-β3 attenuated the inhibitory effect of UA on the proliferation of HCT1 1 6 cells,while the TGF-β3 inhibitor potentiated this effect. UA sup-pressed the transconduction of Wnt/β-catenin signaling in HCT1 1 6 cells through decreasing the level of β-catenin.Exogenous expression of TGF-β3 increased the level of β-catenin and partly reversed the UA-in-duced decrease of β-catenin.However,TGF-β3 inhib-itor potentiated the inhibitory effect of UA on β-catenin in HCT1 1 6 cells.Conclusion The anti-proliferation activity of UA in colon cancer may be partly mediated through down-regulating TGF-β3 to suppress Wnt/β-catenin signaling at least.

Effects of electroacupuncture treatment of different intensities on learning-memory function and expression of β-amyloid protein 1-40 in hippocampus CA1 region in rats with vascular dementia / 中国脑血管病杂志

Objectives To observe the effects of electroacupuncture (EA) treatment of different intensities on learning-memory function and expression of β-amyloid protein 1-40 (Aβ1-40) in hippocampus CA1 region in rats with vascular dementia (VD) and to seek the best intensity of EA treatment.Methods A total of 60 male SD rats (SPF grade) were enrolled.Eight rats were selected as sham operation group with random number table method;others were used to copy the VD model with the modified four-vessel occlusion (4-VO) method.According to the random number table method,the successful model of the rats (n=24) were completely randomly divided into a model group,a 1 am EA group (frequency,2/15 Hz,intensity,1 mA,needle retention time,20 min),and a 3 mA EA group (frequency,2/15 Hz,intensity,3 mA,needle retention time,20 min;n=8 in each group).DU20 (BaiHui) and DU14 (DaZhui) in the EA group were acupunctured once a day for 10 d,and took a rest for 2 d as 1 treatment course.After 2 treatment courses,Morris water maze test was used to detect the ability of learning and memory of rats in each group.Fluorescent quantitative polymerase chain reaction was used to detect the expression level of Aβ1-40 mRNA.Results The mean escape latencies of the water maze test from 2 to 5 days in the sham operation group,model group,1 mA EA group,and 3 mA EA group were 46.8±1.9,40.6±2.3,24.6±1.5,19.4 ±1.2 s;56.3±3.5,51.2±2.6,45.9±2.1,40.8±1.4 s,52.7±1.5,46.0±2.3,31.3±1.2,27.7±1.6 s;and 50.8±3.9,41.5±2.1,29.0±1.1,25.6±1.3 s,respectively;the first time across the original platform were 23.3±1.6,53.9±1.3,30.2±1.4,and 28.1±0.8 s,respectively;the first across the original platform within 120 s were 9.4±0.9,2.6±0.5,6.4±0.7,and 7.2±0.9,respectively;the expression levels of Aβ 1-40 mRNAs in the CA1 regions were 17.3±1.1,40.7±1.1,24.0±1.7,and 22.4±1.8,respectively.There were significant differences among the groups (the F values were 195.88,861.605,103.876,and 380.609,respectively;all P<0.01).Compared with the model group,the mean escape latencies,the first time across the original platform were reduced significantly.Compared with the model group,the times across the original platform within 120 s were increased significantly.Compared with the model group,the gene expression level of Aβ 1-40 mRNA in the center of CA1 region was decreased significantly,and the 3 mA EA group was significantly superior to that of the 1 mA EA group.The difference was statistically significant (all P<0.05).Conclusion EA may improve the learning and memory ability in VD rats and lower the expression level of Aβ 1-40 mRNA in the CA1 region of hippocampus.Effect of 3 mA EA is better than that of the 1 mA EA.

Relationship between anti-proliferationeffect of tetrandrine and TGF-β1 in human colon cancer cells / 中国药理学通报

Aim To study the relationship between the anti-proliferation effect of tetrandrine(Tet) and TGF-β1 in human colon cancer cells.Methods Cell viability assay, Western blot, flow cytometry and Annexin Ⅴ-EGFP staining were introduced to analyze the anti-cancer effect of Tet on HCT116 cells.Real-time PCR, Western blot,cell viability and immunofluorescent were employed to determine the relationship between the anti-cancer effect of Tet and TGF-β1 in HCT116 cells, the relationship between the anti-cancer effect of Tet and PI3K/Akt on HCT116 cells, and how TGF-β1 mediated the anti-cancer effect of Tet on HCT116 cells.Results Compared with the control groups, Tet apparently inhibited the proliferation, and induced cell cycle arrest at G1 phase and apoptosis in HCT116 cells.Tet greatly up-regulated the expression of TGF-β1 either the mRNA or protein level, and exogenous expression of TGF-β1 potentiated the anti-cancer effect of Tet in HCT116 cells, while TGF-β1 inhibitor attenuated it notably.Tet decreased the phosphorylation of Akt1/2/3, but no apparent effect was observed on total protein level of Akt1/2;PI3K inhibitor enhanced the anti-cancer effect of Tet in HCT116 substantially.Exogenous expression of TGF-β1 enhanced the Tet-induced decrease phorphorylation of Akt1/2/3, which was partly reversed by TGF-β1 inhibitor in HCT116 cells.Meanwhile, knockdown of PTEN elevated the level of phorphorylated Akt1/2/3, which was abolished by the exogenous expression of TGF-β1 in HCT116 cells.Conclusion Tet may be a potent candidate drug for colon cancer treatment, and the anti-cancer effect of Tet may be partly mediated by up-regulating TGF-β1 to inactivate PI3K/Akt signal.

Correlation of the levels of serum betatrophin, adiponectin, and interleukin-1β in newly diagnosed type 2 diabetes with different stages of urinary albumin / 中华内分泌代谢杂志

267 newly-diagnosed patients with type 2 diabetes were divided into normoalbuminuria group [group N-UAlb, urinary albumin to creatinine ratio (UACR)300 mg/g, n=76). The control group(group NC) consisted of 114 healthy individuals. Serum betatrophin, adiponectin(APN), and interleukin-1β( IL-1β) levels were determined with ELISA methods and the parameters of body mass index (BMI), estimated glomerular filtration rate(eGFR), HbA1C, fasting plasma glucose(FPG), OGTT 2h plasma glucose(2hPG), fasting insulin(FINS), OGTT 2h postprandial insulin(2hPINS), fasting C-peptide(FCP), homeostasis model assessment insulin resistant index(HOMA-IR), and blood lipid were collected. Compared with group NC, the serum betatrophin levels in patients with type 2 diabetes were obviously increased. In patients with type 2 diabetes, betatrophin levels increased along with the increase of UACR and there were significant differences in betatrophin among the three groups(P<0. 01). Betatrophin positively correlated with UACR, HbA1C, FPG, 2hPG, FINS, 2hPINS, HOMA-IR, TC, LDL-C, and TG( r were 0. 785, 0. 225, 0. 136, 0. 241, 0. 386, 0. 223, 0. 411, 0.216,0.193,and0.298,allP<0.05),and betatrophin were also positively correlated with APN and IL-1β(rwere 0. 643 and 0. 710, both P<0. 01). Stepwise multiple regression analysis showed that UACR, HbA1C, FINS, and TG were independent relevant factors affecting betatrophin levels.