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ObjectiveAgrobacterium tumefaciens-mediated transformation （ATMT） of Clonostachys rosea， an endophytic fungus of Glycyrrhiza uralensis seeds， was established and optimized， and orthogonal test was designed to optimize the colonization conditions of C. rosea for G. uralensis seeds， so as to lay foundation for the development of biofertilizer and the breeding of high-quality G. uralensis. MethodThe conditions of ATMT were optimized from three aspects， including the concentration of acetosyringone， co-culture time and the concentration of conidia of recipient fungi. Then， high-quality transformants were selected. Orthogonal test was used to optimize the colonization conditions by taking co-culture temperature， co-culture time and spore concentration as factors and colonization rate as index. ResultWhen spore concentration was 1×107 cfu·mL-1， acetosyringone concentration was 150 μmol·L-1 and the co-culture time was 60 h， the transformation efficiency of C. rosea was the highest， which was 135 transformants per 1×107 recipient fungal spores. The accuracy and stability of the transformations were tested by cloning the marker gene green fluorescent protein （GFP） and β-glucuronidase （GUS） staining. When co-culture temperature was 25 ℃， co-culture time was 36 h and the spore concentration was 1×106 cfu·mL-1， the colonizing rate for C. rosea back dyeing into G. uralensis seeds by seed soaking method was the highest， which was 71.11%. ConclusionThis study successfully establishes stable and efficient technical systems not only of ATMT in C. rosea， but also of colonization of the transformants into G. uralensis seeds， which can lay a foundation for the development of biofertilizer of G. uralensis.
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OBJECTIVE： To investigate the effects of crystal form on in vivo and in vitro behavior of Astilbin nanosuspensions （AT-NS）. METHODS： AT-NS1 and AT-NS2 were prepared by precipitation method and miniaturized media milling method respectively. The particle size and polydispersity index （PDI） were determined by laser particle size analyzer. X-ray diffraction （XRD）， scanning electron microscopy （SEM）， HPLC and paddle method were used to analyze and compare the structure characteristics， appearance morphology and in vitro dissolution of AT raw material， AT-NS1 and AT-NS2. Totally 15 healthy male SD rats were randomly divided into AT raw material， AT-NS1 and AT-NS2 group， with 5 rats in each group. They were given relevant medicine suspension 120 mg/kg （using water as solvent） intragastrically； blood samples were collected from orbit before medication （0 min） and 5， 10， 20， 30， 60， 120， 240， 480 min after medication. Using rutin as internal standard， HPLC method was used to determine plasma concentration of AT in rats. Pharmacokinetic parameters were calculated by using DAS 2.0 software and then compared. RESULTS： The particle sizes of AT-NS1 and AT-NS2 were （212.48±0.32） nm and （226.36±2.29） nm， respectively； PDI were 0.129 3±0.026 3 and 0.254 7±0.012 4. XRD analysis showed AT-NS1 was amorphous， and AT-NS2 was crystalline. Diffraction peaks of both were different from those of AT raw material. SEM analysis showed that AT-NS1 and AT-NS2 were similar in morphology， and they were spherical and uniform in size； AT raw material was lump with large particle size and different sizes. Results of dissolution tests showed that accumulative dissolution of AT raw material， AT-NS1 and AT-NS2 were 4.54％， 35.01％， 12.22％ at 1 h； accumulative dissolution of them were 24.01％， 81.14％， 64.69％ at 12 h； accumulative dissolution of them were 36.04％， 84.47％， 85.86％ at 24 h， respectively. Results of pharmacokinetic study showed， compared with AT raw material group， cmax and AUC0-∞ of AT-NS1 and AT-NS2 groups as well as t1/2z of AT-NS1 group were increased significantly， while tmax of AT-NS1 group was significantly reduced significantly （P＜0.05）. Compared with AT-NS2 goup， cmax， AUC0-∞ and t1/2z of AT-NS1 group were increased significantly， while tmax was reduced significantly （P＜0.05）. CONCLUSIONS： When AT is prepared into NS， dissolution in vitro and oral absorption in vivo of AT are increased significantly. In a short time， the dissolution/absorption of amorphous NS is faster than crystalline NS.
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OBJECTIVE: To explore the effect of premenstrual syndrome( PMS) on the quality of life( QOL) of female medical staffs. METHODS: By convenient sampling method,1 007 female medical workers from three Grade A class 3 hospitals were selected as study subjects and investigated by PMS Scale,World Health Organization Quality of Life Assessment Instrument Brief Version,Simplified Coping Style Questionnaire and Social Support Rating Scale. RESULTS: The detection rate of PMS among medical staffs was 52. 0%,and the total score of QOL of PMS medical staffs was lower than that of non-PMS medical staffs [( 84. 3 ± 12. 4) vs( 90. 5 ± 11. 6),P < 0. 01]. The multivariate linear logistic regression analysis results showed that the PMS medical staffs who often stay up late,with higher degree of dysmenorrhea,higher job stress,lower positive coping style score,higher negative coping style score,and poor support have lower QOL( P < 0. 05). The coping style and social support of PMS medical staffs can directly affect the QOL. The social support can also indirectly affect the QOL through coping style. CONCLUSION: Coping style is a mediator of social support and QOL.Adopting coping style can improve the QOL of PMS medical staffs.
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Objective: To explore the postoperative effect, prognosis, and prognostic factors for benign testicular tumors. Methods: We retrospectively analyzed the clinical data of 35 patients with benign testicular tumors between May 2004 and May 2017 from Sun Yat-sen University Cancer Center, and the patients were followed up until October 2017. Results: The mean age of the 35 patients was 18.8 (0.4-44.0) years. Among them, 14 patients (40.0%) underwent testis-sparing surgery and 21 (60.0%) underwent radical orchiecto-my, and the tumor sizes were 1.8 (0.4-4.0) cm and 2.7 (1.0-8.0) cm, respectively. All patients had been cured without obvious perioper-ative complications. Postoperative histopathological tumor types included 18 epidermal cysts, 10 mature teratomas, 4 interstitial cell tumors, and 3 adenomatoid tumors. Frozen section examination of 10 cases had been operated, and all results were consistent with paraffin pathology. No patient who underwent testis-sparing surgery showed recurrence and/or metastasis during follow-up, and their sexual function and fertility were well preserved. Conclusions: Testis-sparing surgery is reliable, and the size of the tumor determines its implementation. An intraoperative rapid frozen section examination should be performed in patients with testicular neoplasms of a benign or variable nature diagnosed before operation. Patients with benign testicular tumors should undergo testis-sparing surgery, whereas others should undergo radical orchiectomy.
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Objective To investigate the changes of A20 expression stimulated by free fatty acids (FFA) and its pathway.Methods HepG2 cells and U937 cells were stimulated by 0.5 mmol/L mixed FFA.The expression of A20,phosphor-p65 and phosphor-IκBα of neclear factor (NF)-κB pathway and phosphor-c-Jun N-terminal kinase (JNK),JNK,phosphor-extracellular signal-regulated kinase (ERK),ERK,phosphor-p38 and p38 of mitogen-activated protein kinase (MAPK) pathway were detected by Western blotting.The level of interleukin (IL)-12p,IL-1β,tumor necrosis factor (TNF)-α,IL-6,IL-10 and IL-8 cytokines in the supernatant of cell culture were detected by flow cytometry.T-test was performed for statistical analysis.Results The level of A20 changed along with the stimulated time of FFA.NF-κB and MAPK pathways were activated after FFA stimulation.The secretion of IL-6 and IL-8 increased after HepG2 cells stimulated by FFA and both reached peak at 24 hour.Compared with control group,the difference in IL-8 was statistically significant ((423.8 ± 8.9) pg/mL vs (12.4 ± 4.5) pg/mL,t=41.28,P＜0.01).The difference in IL-6 was also statistically significant ((4 082±423.6) pg/mL vs (52.9±29.5) pg/mL,t=9.49,P＜0.01).After U937 cells were stimulated by FFA,the secretion of IL-8 increased compared with control group.And in a certain period of time the secretion was time dependence.The maximum secretion of 24 hours was (200.6±5.7) pg/mL vs (5.0±3.9) pg/mL,and the difference was statistically significant (t=28.16,P＜0.01).IL-10,IL-12p,IL-1β and TNF-α were detected.Both NF-κB pathway and MAPK pathway were detected.Conclusions The in vitro FFA mediated steatotic cell model could induce the expression change of A20 and secretion of inflammatory cytokines.NF-κB and MAPK pathways involved in the response to FFA in HepG2 cells and U937 cells.
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Objective To study the effect of in utero exposure of TCDD on synthesis of EGF and EGFR in submandibular gland of rat offspring. Methods SD rats were treated with TCDD 5??g/kg on Day 15 of gestation, the expression of EGF and EGFR in submandibular gland of rat offspring were analyzed by immunohistochemistry method. Results On PND32 and PND49, the expression of EGF and EGFR in TCDD experiment group were more than that of control group, the difference was statistically significant.Conclusion In utero exposure of TCDD can promote the synthesis of EGF and EGFR in submandibular gland of rat offspring from prepuberty stage through puberty stage. [