High-level Expression and Purification of Human Zona Pellucida huZP3a^(22-176) and huZP3b^(177-348) Peptides in Escherichia coli

Objective To try making huZP3a^22-176 and huZP3b^177-348 polypeptides （representing an intact huZP^322-348 protein without its N-terminal signal peptide and C-terminal transmembrane domain ） express in E. coli at a higher level Methods The cDNAs encoding huZP3a and huZP3b were obtained with PCR method. The pBV221 plasmid was used to construct thermo-inducible recombinant expression vector. Purification of two target expression products employed an improved method of preparative gel polyacrylamide gel electrophoresis. Results Two polypeptides of recombinant huZP3a （rhuZP3a） and recombinant huZP3b （rhuZP3b） were all expressed respectively in an E. coli BL21（DE3）pLysS strain at a higher level, which were recognized by two specific polyclonal antisera in Western blotting test which recognize a linear B cell epitope present in rhuZP3a or rhuZP3b respectively. Using the shake-flask method, approximately 5 mg of rhuZP3a and rhuZP3b with more than 95% relative homogeneity were harvested from 1 L culture respectively. Conclusion The availability of two rhuZP3 polypeptides will help in detecting the immunogenicities of rhuZP3a and rhuZP3b through animal experiments and confirming the function domain of non-glycosylated huZP3 to induce acrosome reaction in vitro.

Effect of reduced glutathione assisted therapy on immune function, antioxidant function and inflammatory factor in ovarian cancer after chemotherapy

Objective:To investigate the effect of reduced glutathione adjuvant treatment on immune function, antioxidant function and inflammatory cytokines in ovarian cancer patients after chemotherapy.Result:A total of 100 patients with ovarian cancer who were admitted in our hospital from January 2015 to June 2017. According to the enrollment serial number, divided them into research group and control group each 50 cases, the control group was given conventional chemical drug treatment, the research group were given reduced glutathione adjuvant therapy on the basis of chemical therapy. The T lymphocyte subsets, antioxidant indexes MDA, SOD, T-AOC and inflammatory cytokines TNF-α, IL-6, IL-8 levels change in two groups before chemotherapy (T0), 1 month after chemotherapy (T1) and 3 months (T2) were observed.Result: (1) The CD3+,CD4+,CD4+/CD8+ levels at T1 was decreased than T0, and T2 was significantly increased when compared with T1, The CD8+ level at T1 was higher than T0, and the T2 was significantly decreased than T1 in both groups. The levels of the CD3+, CD4+, CD4+/CD8 at T1, T2 in the research group were higher than the control group, while the levels of CD8+ in the research group was lower than the control group. The difference was statistically significant. (2) The level of MDA at T1 was higher than T0, T2 was significantly lower than T1 in both groups. The levels of SOD and T-AOC at T1 were decreased than T0, and T2 was significantly higher than T1. The levels of SOD and T-AOC at T1, T2 in the research group were higher than in the control group, while the MDA level was lower than in the control group. The difference was statistically significant. (3) The levels of TNF-α, IL-6 and IL-8 at T1 in two groups were significantly higher than T0, and the T2 decreased significantly when compared with T1, The levels of TNF-α, IL-6 and IL-8 at T1, T2 in the research group were lower than in the control group. The difference was statistically significant.Conclusion: The reduced glutathione adjuvant chemotherapy for postoperative ovarian cancer can help to protect the body's immune function, antioxidant function and reduce inflammation factor.

Field responses of the Asian larch bark beetle, Ips subelongatus, to potential aggregation pheromone components＂ disparity between two populations in northeastern China

Behavioral responses of the Asian larch bark beetle, lps subelongatus Motsch. to three potential aggregation pheromone components, ipsenol （racemic or [-]- enantiomer）, ipsdienol （racemic or [＋]-enantiomer） and 3-methyl-3-buten- l-ol, were tested using partial or full factorial experimental designs in two provinces （Inner Mongolia and Jilin） of northeastern China. Our field bioassays in Inner Mongolia （Larix principis- rupprechtii Mayr. plantation） clearly showed that ipsenol, either racemic or 97%-（-）- enantiomer, was the only compound that significantly attracted both sexes of 1. subelongatus, while all other compounds （singly or in combinations） were unattractive. There were no two- or three-way synergistic interactions. However, in Jilin Province （L. gmelini [Rupr.] Rupr. Plantation）, all the individual compounds tested were inactive, except a very weak activity by 97%-（-）-ipsenol in 2004 when the beetle population was very high. While a combination of ipsenol and ipsdienol （racemates or enantiomerically pure natural enantiomers） showed a significant attraction for both sexes of I. subelongatus, indicat- ing a two-way synergistic interaction between these two major components, addition of 3-methyl-3-buten-l-ol to these active binary blend（s） did not have any effects on trap catches, suggesting that ipsenol and ipsdienol are the synergistic aggregation pheromone components of I. subelongatus in Jilin Province. It seems that 97%-（-）-ipsenol in Inner Mongolia or the binary blend of 97%-（-）-ipsenol and 97%-（＋）-ipsdienol in Jilin Province are superior to their corresponding racemates, which might be due either to weak inhibitory effects of the antipode enantiomers or to reduced release rates of the active natural enan- tiomer（s） in the racemate（s）. Our current bioassay results suggest that there is a strong geographical variation in aggregation pheromone response of L subelongatus in northeastern China. Future research on the pheromone production and response of 1. subelongatus from different regions in Northeast Asia will surely improve our understanding of the dynamic aggregation pheromone system of this economically important forest pest insect.