OBJECTIVE To investigate the potential mechanism of epigallocatechin gallate （EGCG） enhancing the sensitivity of hepatocellular carcinoma （HCC） cells to lenvatinib based on the phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. METHODS Five human HCC cell lines （HepG2， Huh-7， SMMC-7721， SNU-368 and SNU-739） were used to evaluate the effects of lenvatinib alone and in combination with EGCG on survival rates， clone number， proliferation rate， invasion number and the expressions of mRNAs and proteins related to the PI3K/Akt signaling pathway. The PI3K activator insulin-like growth factor-1 （IGF-1） was introduced to investigate the effect of activating the PI3K/Akt signaling pathway on the sensitization effect of EGCG. RESULTS Compared with the control group， lenvatinib （10 μmol/L） and different concentrations of EGCG+ lenvatinib （1， 5 and 10 μg/mL EGCG+10 μmol/L lenvatinib） significantly reduced the survival rates and clone numbers of all five HCC cell lines in a dose-dependent manner （P＜0.05）. Lenvatinib （10 μmol/L） and EGCG+lenvatinib （10 μg/mL EGCG+10 μmol/L lenvatinib） also markedly inhibited the proliferation rate and invasion numbers of these cells， and decreased the mRNA expressions of PI3K， Akt， mammalian target of rapamycin （mTOR）， P70S6K and 4EBP， and the phosphorylation levels of PI3K and Akt， as well as the protein expressions of mTOR and B cell lymphoma-2 （Bcl-2） in HepG2 cells or all five HCC cells； conversely， the mRNA and protein expressions of phosphatase and tensin homologue deleted on chromosome 10（PTEN）， and the protein expressions of caspase-3 and cleaved caspase-3 were significantly upregulated， with more pronounced effects observed in the EGCG+lenvatinib group than in the lenvatinib group （P＜0.05）. Compared with the lenvatinib group and the EGCG+lenvatinib group， the clone number， proliferation rate and invasion number of HepG2 cells in the EGCG+lenvatinib+IGF-1 group （10 μg/mL EGCG+10 μmol/L lenvatinib+50 ng/mL IGF-1） were significantly increased （P＜0.05）. CONCLUSIONS EGCG can enhance the sensitivity of HCC cells to lenvatinib， and its underlying mechanism may be related to the inhibition of the activation of PI3K/Akt signaling pathway activation.

Improving the nitrogen use efficiency (NUE) of Brassica napus is of significant importance for achieving the national goal of zero growth in chemical fertilizer application and ensuring the green development of the rapeseed industry. This study aims to explore the effects of the nitrate transporter gene BnaNRT1.5s on the nitrogen transport and NUE of B. napus, providing excellent genetic resources for the development of nitrogen-efficient B. napus varieties. The spatiotemporal expression of BnaA05.NRT1.5 as a key nitrogen responsive gene was profiled by qRT-PCR at different growth stages and for different tissue samples of B. napus 'Westar'. Subcellular localization was employed to examine its expression pattern in the cells. Additionally, CRISPR/Cas9 was used to create BnaNRT1.5s knockout lines, which were subjected to hydroponic experiments under high nitrogen (12.0 mmol/L) and low nitrogen (0.3 mmol/L) conditions. After the seedlings were cultivated for 21 days, root and shoot samples were collected for weighing, nitrogen content determination, xylem sap nitrate content assessment, and calculation of total nitrogen and NUE. The B. napus nitrate transporter BnaA05.NRT1.5 was localized to the cell membrane. During the seedling and early bolting stages, BnaA05.NRT1.5 was predominantly expressed in roots, while it was highly expressed in old leaves and mature silique skin during the reproductive stage. Compared with the wild type, the mutant BnaNRT1.5s showed significant increases in the dry weight and total nitrogen of seedlings under both high and low nitrogen conditions. Under low nitrogen conditions, NUE in the roots of BnaNRT1.5s significantly improved. Notably, under both high and low nitrogen conditions, the nitrate content in the shoots of BnaNRT1.5s decreased significantly, while that in the roots increased significantly, resulting in a significantly decreased shoot-to-root nitrate content ratio. BnaNRT1.5s is involved in regulating the transport of nitrate from the roots to the shoots, and its mutation enhances nitrogen absorption and utilization in B. napus seedlings, promoting seedling growth. This study not only provides references for understanding the physiological and molecular mechanisms by which BnaNRT1.5s regulates NUE but also offers valuable genetic resources for improving NUE in B. napus.
Brassica napus/genetics* ; Anion Transport Proteins/metabolism* ; Nitrogen/metabolism* ; Nitrate Transporters ; Plant Proteins/metabolism* ; Nitrates/metabolism* ; Gene Expression Regulation, Plant ; Biological Transport

OBJECTIVE:To study the effects of ginsenoside CK combined with 5-fluorouracil (5-FU) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of human pancreatic cancer PANC-1 cells. METHODS:PANC-1 cells of logarithmic growth phase were randomly divided into blank control group,ginsenoside CK group (30 mg/L),5-FU group (25 mg/L)and combination group(ginsenoside CK 30 mg/L+5-FU 25 mg/L). MTT method was used to detect the cell proliferation in-hibition rate in each group after 24,48,72 h;flow cytometry was used to detect the cell apoptosis rate after 48 h;enzyme-linked immunosorbent assay was used to detect the fibronectin expression in cells after 24,48,72,96 h;and Western blot was used to detect the expressions of vimentin,N-cadherin,E-cadherin,protein kinase(Akt)and phosphorylated Akt(p-Akt)protein in cells after 48 h. RESULTS:Compared with blank control group,the cell proliferation inhibition rate,early and late apoptotic rates,pro-tein expression level of E-cadherin in ginsenoside CK group,5-FU group and combination group were obviously increased (P<0.05),while the protein expression levels of fibronectin,vimentin,N-cadherin,and p-Akt/Akt levels were obviously decreased (P<0.05);the effects of above-mentioned indexes in combination group were superior to ginsenoside CK group and 5-FU group (P<0.05). CONCLUSIONS:Both ginsenoside CK and 5-FU can inhibit the proliferation of PANC-1 cells,induce their apoptosis and inhibit EMT,which may be associated with inhibiting phosphatidylinositol 3-kinase/Akt pathway. In addition,the combination of ginsenoside CK and 5-FU can produce a better effect.

OBJECTIVE:To observe clinical efficacy and safety of Astragalus injection combined with neoadjuvant chemother-apy and three dimensional conformal radiotherapy in the treatment of colorectal cancer. METHODS:A total of 80 patients with me-dium and advanced colorectal cancer were selected from oncology department of our hospital during Jun. 2010 to Jun. 2014,and di-vided into observation group and control group according to random number table,with 40 cases in each group. Control group re-ceived neoadjuvant chemotherapy combined with three-dimensional conformal radiotherapy. Observation group was additionally giv-en Astragalus injection 10-20 mL added into 5％ Glucose injection 250-500 mL,ivgtt,qd,on the basis of control group. A treat-ment course lasted for 21 d,and both groups received 3 courses. Short-term efficacies as well as immune indexes [CD3+,CD4+, CD4+/CD8+ and NK cells] before and after treatment were observed in 2 groups. The occurrence of ADR and survival situation were compared between 2 groups. RESULTS:Clinical response rate of observation group was 87.5％,which was slightly higher than that of control group(77.5％),but there was no statistical significance(P>0.05). Before treatment,there was no statistical signifi-cance in immune index levels between 2 groups(P>0.05). After treatment,NK cells,CD3+,CD4+ and CD4+/CD8+ levels of con-trol group were decreased significantly,which was significantly lower than observation group,with statistical significance (P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). Two-year survival rate of observa-tion group was significantly higher than that of control group;the recurrence rate and distant metastasis rate were significantly low-er than control group,with statistical significance(P<0.05). CONCLUSIONS:Astragalus injection not only can improve therapeu-tic efficacy of neoadjuvant chemotherapy in patients with colorectal cancer,balance inflammatory state,improve survival rate,re-duce recurrence rate and distant metastasis rate with good safety.