This study aims to investigate the role of slient mating-type information regulation 2 homolog 1(SIRT1)/tuberous sclerosis complex 2(TSC2)/mammalian target of rapamycin(mTOR) signaling pathways in the Periplaneta americana extract CⅡ-3-induced senescence of human leukemia K562 cells. K562 cells were cultured in vitro and treated with 0(control), 5, 10, 20, 40, 80, and 160 μg·mL~(-1) of P. americana extract CⅡ-3. Cell counting kit-8(CCK-8) and flow cytometry were employed to examine the proliferation and cell cycle of the K562 cells. Senescence-associated β-galactosidase stain kit(SA-β-gal) was used to detect the positive rate of senescent cells. Mitochondrial membrane potential was detected by flow cytometry. The relative mRNA level of telomerase reverse transcriptase(TERT) was determined by fluorescence quantitative PCR. The mRNA and protein levels of SIRT1, TSC2, and mTOR were determined by fluorescence quantitative PCR and Western blot, respectively. The results showed that CⅡ-3 significantly inhibited the proliferation of K562 cells and the treatment with 80 μg·mL~(-1) CⅡ-3 for 72 h had the highest inhibition rate. Therefore, 80 μg·mL~(-1) CⅡ-3 treatment for 72 h was selected as the standard for subsequent experiments. Compared with the control group, CⅡ-3 increased the proportion of cells arrested in G_0/G_1 phase, decreased the proportion of cells in S phase, increased the positive rate of SA-β-Gal staining, elevated the mitochondrial membrane potential and down-regulated the mRNA expression of TERT. Furthermore, the mRNA expression of SIRT1 and TSC2 was down-regulated, while the mRNA expression of mTOR was up-regulated. The protein expression of SIRT1 and p-TSC2 was down-regulated, while the protein expression of p-mTOR was up-regulated. The results indicated that P. americana extract CⅡ-3 induced the senescence of K562 cells via the SIRT1/mTOR signaling pathway.
Humans ; Animals ; Periplaneta ; Sirtuin 1/genetics* ; K562 Cells ; Signal Transduction ; TOR Serine-Threonine Kinases/genetics* ; RNA, Messenger ; Mammals

Objective: To investigate the effect of the Periplaneta Americana polypeptide on the angiogenesis. Method:Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell scratch assay were used to observe effect of different concentration (6.25,12.5,25,50,100 mg·L^-1) of the Periplaneta Americana polypeptide, CⅡ-3 and skimmed cream on the proliferation and migration of human umbilical vein endothelial cells (HUVECs), and a normal group and a thalidomide group were also established in this study. The tubule formation assay was used to detect the effect of different concentration (25, 50, 100 mg·L^-1) of the Periplaneta Americana extracts on the formation of tubules in HUVECs cells. The adhesion between HepG2 cells and HUVECs cells was observed by cell adhesion assay. The expression of vascular endothelial growth factor (VEGF) proteins in HUVECs was detected by immunocytochemical staining and enzyme linked immunosorbent assay (ELISA). Result:MTT results showed that the Periplaneta Americana polypeptide could inhibit the proliferation of HUVECs in a dose-dependent manner (P<0.05). The effect of different concentrations of PAP-2 was better than that of PAP-1 and PAP-3 at 24, 48 72 h (P<0.05). However, the survival rate of HUVECs was significantly increased after treatment with different concentrations of CⅡ-3 and skimmed cream in a time-dose dependent manner. Cell wound scratch assay indicated that migration of HUVECs could be inhibited by Periplaneta Americana polypeptide in a dose-dependent manner (P<0.05), and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). CⅡ-3 could inhibit migration HUVECs to some extent, but the higher the dose was, the weaker the inhibition effect was. Skimmed cream promoted migration of HUVECs in a dose-dependent manner. Tube formation assay revealed that Periplaneta Americana polypeptide could inhibit tube formation of HUVECs, and the inhibitory effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). CⅡ-3 and skimmed cream promoted the tube formation of HUVECs to a certain extent. Intercellular adhesion assay stated clearly that Periplaneta Americana polypeptide could block the adhesion between HepG2 and HUVECs (P<0.05), and the effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). However, CⅡ-3 and skimmed cream could promote the adhesion between HepG2 and HUVECs. Results of ELISA assay and immunocytochemical staining indicated that Periplaneta Americana polypeptide could down-regulate the expression of VEGF of HUVECs in a dose-dependent manner (P<0.05), in which effect of PAP-2 was better than that of PAP-1 and PAP-3 (P<0.05). However, CⅡ-3 and skimmed cream could up-regulate the expression of VEGF in HUVECs. Conclusion:The Periplaneta Americana polypeptide can inhibit the invasion, metastasis and tube formation of HUVECs, and down-regulate the expression of VEGF in HUVECs. The effect of Periplaneta Americana polypeptide is better than CⅡ-3 and skimmed cream, and the among the polypeptide, the effect of PAP-2 is superior to the other two.

Objective To study the inhibitory effect of T lymphocytes secreting EphrinAl-Caspase-3 in vivo and on the growth of cancer cells in nude mice with breast cancer. Methods Nude mice (n = 35) were inoculated with breast cancer cells to construct a nude mouse model of breast cancer. When the tumor volume reached 0. 1 cm3, 30 nude mice with average size tumor tissue were randomly divided into PBS group, uninfected adenovirus group, T lymphocyte infected with Ad-EphrinAl-Caspase-3 group, and intratumoral transplantation. Tumor size was measured every day 2 to 3. Three groups of tumor-bearing nude mice were selected. After the above-mentioned cell transplantation, the subcutaneous tumor tissue homogenate was obtained every day 2 to 3, and the content of EphrinAl-Caspase-3 was detected by ELISA. At the end of the experiment, the animals in each group were sacrificed by cervical dissection and sliced. The presence of T lymphocytes expressing green fluorescent protein was observed under a fluorescence microscope, and Caspase-3 and Ki-67 were detected by immunofluorescence. Results After one week of inoculation of breast cancer cells into nude mice, the presence of subcutaneous tumors could be touched by hand, which proved that the tumor-bearing animals of breast cancer cells were successfully modeled. On the 8th day after inoculation, the tumor volume of the nude mice in each group became larger, and the difference between the treatment group and the PBS group/T lymphocyte group was extremely significant ( P<0.05). Although the tumor volume of the T lymphocyte transplantation group was slower than that of the PBS control group, there was no statistically significant difference between the two. The expression of EphrinAl-Caspase-3 was detected in the EphrinAl-Caspase-3 treatment group on the 2nd day, reached the peak on the 8th day, and then the secretion decreased gradually. No expression of EphrinAl-Caspase-3 was detected in the PBS control group and the T lymphocyte group. The presence of dispersed green fluorescent protein-labeled EphrinAl-Caspase-3-T lymphocytes was observed in the tumor tissues of the treatment group, while the presence of green fluorescent protein was not detected in the PBS group and the T lymphocyte groups. In the infected cells of the treatment group, the proportion of Caspase-3 positive cells was up- regulated, and the proportion of Ki-67 positive cells was down-regulated. No expression of EphrinAl-Caspase-3 was detected in the PBS group and the T lymphocyte group. Conclusion EphrinAl-Caspase-3 can significantly inhibit the growth of breast cancer cells, thereby exerting an anti-tumor effect.

A novel series of benzyl urea analogues based on the structural modification of sorafenib were synthesized. Their in vitro antitumor activities against MX-1, HepG2, Ketr3 and HT-29 were evaluated using the standard MTT assay. While several target compounds showed inhibitory activity against multiple cancer cell lines, compound 9 was of particular interest, demonstrating IC50 values (5.69-13.6 micromol x L(-1)) comparable to those of sorafenib. Furthermore, compounds 20 and 23 showed more potent inhibitory activity against HT-29 and MX-1 when compared to sorafenib. In particular, compound 20 bearing the N-3-pyridyl moiety not only exhibited greater inhibitory activity against HT-29 cell line (IC50 3.82 micromol x L(-1)), but also had improved solubility at pH 7.2, is worthy of further investigation as a lead to identify novel antitumor agents.

Objective To evaluate the dosimetric imnpact of the fixed position of two-degrade collimator in the treatment of breast cancer after radical mastectomy using intensity-modulated radiotherapy (IMRT) technique.Methods A total of ten patients with breast cancer were treated with radical mastectomy and radiotherapy sequaciously involving the supraclavicular region and the chest wall.Two different IMRT treatment plans were designed for each patient:0°,40° and two tangential field.There was no restriction on the position of two-degrade collimator(IMRT-1) (P ＞ 0.05).The beam angles and the parameters were as same as IMRT-1,but fixed the position of the two-degrade collimator of 0° and 40° at the inferior border of the supraclavicular (IMRT-2).The dose distribution of target volume and normal tissues,conformal index (CI),and heterogeneous index (HI) were estimated with the dose volume histogram (DVH) for the two intensity modulated modes.Results The CI were 0.79 and 0.73 (Z =-2.316,P＜0.05),and the HI of the IMRT-2 plans was not different from IMRT-1 (P ＞ 0.05).Considering the dose volunes of the ipsilateral lung in two plans,the values of V5,V10,D of IMRT-2 were significantly less than those of IMRT-1 (Z =-2.805,-2.812,-2.521,P ＜ 0.05).Meanwhile,the D of the contralateral lung,D of heart and D of the contralateral breast from the IMRT-2 were all lower than those oflMRT-1 (Z=-2.666,-2.701,-2.310,P＜0.05).There was no significant difference in the values of V20,V30 of the ipsilateral lung,V30 of heart and between IMRT-1 and IMRT-2 (P ＞0.05).Conclusions Compared with IMRT-1,IMRT-2 with fixed position of the two-degrade collimator could significantly reduce the low dose region of the lung and heart.It may be used as an effective alternative for breast cancer after radical mastectomy irradiation.

Objective:To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism.Methods:The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells;the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method;the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits.Results:The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation,the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation,the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation.Conclusion:ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

OBJECTIVE:To provide reference for the safe and rational drug use for children. METHODS:Information manage-ment system was used to investigate the use of essential medicines system variety in stock in 2015 and analyze the medication infor-mation for children in the drug instructionsin our hospital in 2015. RESULTS:Only 201 kinds of medicines belonged to children’s medicines in all the 685 kinds of medicines in our hospital. And 89 kinds (44.28%) of medicines belonged to essential medicine system among the 201 kinds of children’s medicines,in which,78 (87.60%) showed complete medication information for chil-dren;112 kinds(55.72%)of medicines belonged to non-essential medicine system,in which,38(33.93%)showed complete medi-cation information for children. The proportions of showing complete medication information for children in the essential medicines and in its chemicals,biological products,injections and oral preparations were higher than non-essential medicines,the differences were statistically significant(P<0.05). Only 41 kinds of medicines belonged to child-specific medicines among the 201 children’s medicines;62 showed complete medication information for children in the 73 kinds of essential medicines among the non-child-spe-cific medicines;only 13 showed complete medication information for children in the 87 kinds of non-essential medicines,the pro-portion of showing complete medication information for children in essential medicines among the non-child-specific medicines was higher than non-essential medicines,the difference was statistically significant(P<0.05). CONCLUSIONS:The use proportion of essential medicine system variety for children’s medicines is high in our hospital;but there are lacking of child-specific medicines and the medication information for children is insufficient. However,compared with non-essential medicines for children,the essen-tial medicines show better medication information for children in aspects of types,dosage form distribution and non-child-specific medicines,and it is suitable for children.

Wnt signaling are critical pathway involved in organ development,tumorigenesis,and cancer progression.WNT7A,a member of the Wnt family,remains poorly understood in terms of its role and the underlying molecular mechanisms it entails in head and neck squamous cell carcinoma(HNSCC).According to the Cancer Genome Atlas(TCGA),transcriptome sequencing data of HNSCC,the expression level of WNT7A in tumors was found to be higher than in adjacent normal tissues,which was validated using Real-time RT-PCR and immunohistochemistry.Unexpectedly,overexpression of WNT7A did not activate the canonical Wnt-β-catenin pathway in HNSCC.Instead,our findings suggested that WNT7A potentially activated the FZD7/JAK1/STAT3 signaling pathway,leading to enhanced cell proliferation,self-renewal,and resistance to apoptosis.Furthermore,in a patient-derived xenograft(PDX)tumor model,high expression of WNT7A and phosphorylated STAT3 was observed,which positively correlated with tumor progression.These findings underscore the significance of WNT7A in HNSCC progression and propose the targeting of key molecules within the FZD7/JAK1/STAT3 pathway as a promising strategy for precise treatment of HNSCC.

Wnt signaling are critical pathway involved in organ development,tumorigenesis,and cancer progression.WNT7A,a member of the Wnt family,remains poorly understood in terms of its role and the underlying molecular mechanisms it entails in head and neck squamous cell carcinoma(HNSCC).According to the Cancer Genome Atlas(TCGA),transcriptome sequencing data of HNSCC,the expression level of WNT7A in tumors was found to be higher than in adjacent normal tissues,which was validated using Real-time RT-PCR and immunohistochemistry.Unexpectedly,overexpression of WNT7A did not activate the canonical Wnt-β-catenin pathway in HNSCC.Instead,our findings suggested that WNT7A potentially activated the FZD7/JAK1/STAT3 signaling pathway,leading to enhanced cell proliferation,self-renewal,and resistance to apoptosis.Furthermore,in a patient-derived xenograft(PDX)tumor model,high expression of WNT7A and phosphorylated STAT3 was observed,which positively correlated with tumor progression.These findings underscore the significance of WNT7A in HNSCC progression and propose the targeting of key molecules within the FZD7/JAK1/STAT3 pathway as a promising strategy for precise treatment of HNSCC.

Wnt signaling are critical pathway involved in organ development,tumorigenesis,and cancer progression.WNT7A,a member of the Wnt family,remains poorly understood in terms of its role and the underlying molecular mechanisms it entails in head and neck squamous cell carcinoma(HNSCC).According to the Cancer Genome Atlas(TCGA),transcriptome sequencing data of HNSCC,the expression level of WNT7A in tumors was found to be higher than in adjacent normal tissues,which was validated using Real-time RT-PCR and immunohistochemistry.Unexpectedly,overexpression of WNT7A did not activate the canonical Wnt-β-catenin pathway in HNSCC.Instead,our findings suggested that WNT7A potentially activated the FZD7/JAK1/STAT3 signaling pathway,leading to enhanced cell proliferation,self-renewal,and resistance to apoptosis.Furthermore,in a patient-derived xenograft(PDX)tumor model,high expression of WNT7A and phosphorylated STAT3 was observed,which positively correlated with tumor progression.These findings underscore the significance of WNT7A in HNSCC progression and propose the targeting of key molecules within the FZD7/JAK1/STAT3 pathway as a promising strategy for precise treatment of HNSCC.