Objective To investigate the apoptotic effect of petasin on myeloma RPMI 8226 cells and the mechanisms.Methods The inhibition of petasin on the proliferation of myeloma RPMI 8226 cells was tested by trypan blue assay.Apoptosis of RPMI 8226 cells was measured by terminal-deoxynueleotidyl transferase mediated dUTP nick end labeling (TUNEL)assay and Hoechst 33258 staining assay.Effects of petasin on caspase-3,8 and 9 expressions,phosphorylation of ERK1/2,MEK(p-ERK1/2 ;p-MEK)and p38MAPK(p-p38MAPK)protein were analyzed by Western blot.Results Incubation by petasin for 24 h,48 h or 72 h could significantly inhibit the pro-liferation of myeloma RPMI 8226 cells (P <0.01,P <0.01,P <0.01).Petasin induced the apoptosis of myeloma RPMI 8226 cells in time-and concentration-dependent manners (P <0.05,P <0.05).Caspase inhibitor pretreat-ment could significantly inhibit the apoptosis of myeloma cells.After cultured with petasin for 72 h,the expressions of caspase-3,8 and 9 were obviously enhanced (P <0.05,P <0.01,P <0.05)and phosphorylation of p-p38MAPK of RPMI8226 cells was significantly increased (P <0.01).However,phosphorylation of p-ERK1/2 and p-MEK was decreased significantly (P <0.01,P <0.05).Conclusion Petasin can inhibit the proliferation of myeloma RPMI 8226 cells and induce apoptosis.The mechanism may be related to the activation of caspase-3,8 and 9 proteins and the changes in phosphorylation of p38MAPK,ERK1/2 and MEK.

Objective To evaluate the efficacy of serum PCT level in deciding the development and progno-sis of sepsis and its conrrelation with APACHE Ⅱ scoring.Methods 56 patients of sepsis accepted intensive care treatment and were all given APACHE Ⅱ scoring within the first 24 h after admission to ICU.The PCT level at dif-ferent time(1 d,3 d,5 d-7 d,10 d after admission)was detected.All these patients were divided into survival group and death group based on the 28-day fatality.Results The PCT level declined gradually with the treatment and it decreased obviously from the third day in comparison with the original level before admission [survival group/death group:(2.98±0.48)μg/L/(4.98±0.66)μg/L vs(4.04±0.50)μg/L/(6.02±0.50)μg/L](P

AIK is a novel cationic peptide with potential antitumor activity. In order to construct the AIK expression vector by Gateway technology, and establish an optimal expression and purification method for recombinant AIK, a set of primers containing AttB sites were designed and used to create the AttB-TEV-FLAG-AIR fusion gene by overlapping PCR. The resulting fusion gene was cloned into the donor vector pDONR223 by attB and attP mediated recombination (BP reaction), then, transferred into the destination vector pDESTl 5 by attL and attR mediated recombination (LR reaction). All the cloning was verified by both colony PCR and DNA sequencing. The BL21 F. coli transformed by the GST-AIR expression plasmid was used to express the GST-AIK fusion protein with IPTG induction and the induction conditions were optimized. GST-AIR fusion protein was purified by glutathione magnetic beads, followed by rTEV cleavage to remove GST tag and MTS assay to test the growth inhibition activity of the recombinant AIR on human leukemia HL-60 cells. We found that a high level of soluble expression of GST-AIK protein (more than 30% out of the total bacterial proteins) was achieved upon 0.1 mmol/L ITPG induction for 4 h at 37 °C in the transformed BL21 F. coli with starting OD₆₀₀ at 1.0. Through GST affinity purification and rTEV cleavage, the purity of the resulting recombinant AIK was greater than 95%. And the MTS assays on HL-60 cells confirmed that the recombinant AIK retains an antitumor activity at a level similar to the chemically synthesized AIK. Taken together, we have established a method for expression and purification of recombinant AIK with a potent activity against tumor cells, which will be beneficial for the large-scale production and application of recombinant AIK in the future.
Antimicrobial Cationic Peptides ; biosynthesis ; Antineoplastic Agents ; metabolism ; Escherichia coli ; metabolism ; Genetic Vectors ; HL-60 Cells ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Sequence Analysis, DNA

Objective To evaluate the clinical effects and safety of paclitaxel liposome or gemcitabine combined with cisplatin in elderly patients with advanced lung squamous cell cancer. Methods A total of 52 elderly patients with advanced lung squamous cell cancer in Kunshan First People's Hospital from January 2012 to October 2016 who received paclitaxel liposome with cisplatin (n=24) or gemcitabine with cisplatin(n = 28) were analyzed retrospectively. The patients received at least 2 cycles of chemotherapy, 2-6 cycles in total. CT was rechecked every 2 cycles. The clinical effects and adverse reactions were evaluated by National Cancer Institute Response Evaluation Criteria in Solid Tumors (NCI RECIST) and NCI Common Terminology Criteria for Adverse Events (NCI CTCAE). Results The objective response rate (ORR) and the disease control rate(DCR)in paclitaxel liposome group and in gemcitabine group had a lot in common [ORR:29.2 %(7/24)vs.32.1 %(9/28),χ2=0.054,P =0.817;DCR:70.8 %(17/24)vs.71.4 %(20/28),χ2=0.002,P =0.962]. The median progression-free survival(PFS) time in gemcitabine group was longer than that in paclitaxel liposome group (5.7 months vs. 5.4 months, χ2= 0.466, P = 0.495). One-year survival rate in gemcitabine group (37.0 %) was higher than that in paclitaxel liposome group (34.8 %) and there was no statistically significant difference (χ 2= 0.027, P = 0.869). However, hematologic adverse reactions in paclitaxel liposome group were lower than those in gemcitabine group (P< 0.05). Conclusion The effect of paclitaxel liposomes combined with cisplatin for treatment of elderly patients with advanced lung squamous cell carcinoma is not inferior to gemcitabine combined with cisplatin, which has less adverse reactions and may be recommended as the first-line chemotherapy for the patients.

It is not fully clear why there is a higher contribution of pluripotent stem cells (PSCs) to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells (ESCs) but also induced pluripotent stem cells (iPSCs) can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a "dose effect". Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast (EPI) lineage development while promoting trophectoderm (TE) differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A (cultured from 4-cell stage to early blastocyst at E3.5) could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improve their EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understanding of PSCs effects on early embryo development.
Activins ; metabolism ; Animals ; Cells, Cultured ; Embryonic Development ; Germ Layers ; metabolism ; Mice ; Pluripotent Stem Cells ; cytology ; metabolism

The fall armyworm (FAW), Spodoptera frugiperda, is a destructive pest native to America and has recently become an invasive insect pest in China. Because of its rapid spread and great risks in China, understanding of FAW genetic background and pesticide resistance is urgent and essential to develop effective management strategies. Here, we assembled a chromosome-level genome of a male FAW (SFynMstLFR) and compared re-sequencing results of the populations from America, Africa, and China. Strain identification of 163 individuals collected from America, Africa and China showed that both C and R strains were found in the American populations, while only C strain was found in the Chinese and African populations. Moreover, population genomics analysis showed that populations from Africa and China have close relationship with significantly genetic differentiation from American populations. Taken together, FAWs invaded into China were most likely originated from Africa. Comparative genomics analysis displayed that the cytochrome p450 gene family is extremely expanded to 425 members in FAW, of which 283 genes are specific to FAW. Treatments of Chinese populations with twenty-three pesticides showed the variant patterns of transcriptome profiles, and several detoxification genes such as AOX, UGT and GST specially responded to the pesticides. These findings will be useful in developing effective strategies for management of FAW in China and other invaded areas.
Animals ; China ; Genomics ; Humans ; Male ; Pesticides ; Spodoptera/genetics* ; Transcriptome