Objective To amplify natural killer (NK) cells in vitro and explore its killing effect on ovarian cancer cells.Methods (1) The separation of NK cells and identification.A total of 20 ml peripheral blood of one healthy volunteer was collected in Nov.2015,Peking University People's Hospital.The peripheral blood mononuclear cells of normal volunteers were isolated,cultured in vitro and amplificated cultivation for 14 days with K562 cells transfected and expressing interleukin 21 (IL-21-K562) as nourish cells.The number and dynamic state of the growth cells were monitored during the cultured process.Cells were harvested and counted after 14 days cultured.The NK cells phenotypes were detected by flow cytometry.(2) The killing effect of NK cells on ovarian cancer cells:the ratio of effector cells (NK cells) and target cells (ovarian cancer cells and its control) was 50∶ 1,20∶ 1,10∶ 1,5∶1 or 1 ∶ 1,NK cells killing effect on ovarian cancer cells was detected by the lactate dehydrogenase (LDH) release experiments.Results (1) The results of NK cells establishment and phenotypic characterization:the cells were induced in vitro for 14 days by amplification culture.With the extension of incubation time,the number of NK cells increased constantly,from 2.0× 107 on day 0 to 5.1 × 109 on day 14.Obvious amplification of the total number of cells were detected for 255 times.Living cells unstained by trypan blue eventually reached 95％ above.Before and after the induction and amplification in vitro,the percentage of NK cells (CD3-CD56+cells) in CD3-cells were 2.33％ and 85.32％,respectively (P＜0.01),which covered the whole lymphocytes 1.06％ and 69.42％,respectively (P＜0.01),which showed that NK was the main cell type in the amplificated lymphocytes.(2) The killing rate of NK cells on ovarian cancer cells in vitro:the results detected by LDH release experiments showed that NK cells could performed strong nonspecific killing effect on ovarian cancer cell lines SKOV3,HOC1A,3AO and CAOV3,as well the normal ovarian cell line T29 and NK sensitive cell line K562,and the killing effect increased significantly along with the increase of effector cells and target cells ratio (P＜0.01).When the ratio was 1 ∶ 1,the killing rate was 37％ for K562,while the rate of killing of other cells was around 10％ (P＜0.05).When the effect-target ratio was 20∶1 and 50∶ 1,in addition to CAOV3 cells (more than 70％),NK cells had a kill rate of more than 80％ for other ovarian cancer cells lines and their control cell K562 and T29 cells (P＞0.05).Conclusion NK cells could be established in vitro and have a good non-specific killing effect on ovarian cancer cells.

Objective To explore changes of three-dimensional organization of collagen fibers framework in hepatic fibrosis rats. Methods Healthy adult male Wistar rats(n=30) were randomly divided into control group(n=10) and hepatic fibrosis group.Hepatic fibrosis rats were induced by subcutaneous injections of 60% carbon tetrachloride oily solution.Rats were killed at the end of 10 week and 12 week as 10week(n=10) and 12week(n=10) hepatic fibrosis group.Distribution of collagen was detected by sirius red and polarized microscope,and the architecture was observed under the scanning electron microscope in hepatic fibrosis rats. Results Under the scanning electron microscope,with aggravation of hepatic fibrosis,kinds of collagen fibers and interlaced reticulation compartment were observed,some round or olivary !honeycomb"-like frameworks were formed,which disparted and wrapped up normal liver lobule,and the collagen fiber sheaths formed the deposition of collagen fibers surrounded portal areas or vessels.Pseudolobule vesicles formed in 12week hepatic fibrosis rats were more delicate and intact than that of in 10week hepatic fibrosis rats. Conclusion The microframework of the collagen fibers was changed in hepatic fibrosis.This structure was mainly composed of thick and slight hyperplastic collagen fibers.;

Objective:To compare the clinical and prognostic characteristics of ovarian endometrioid carcinoma (OEC) patients with synchronous endometrial lesions and patients with pure OEC.Methods:A retrospective review of the medical records of patients received initial treatment and a postoperative pathological diagnosis of OEC at Peking University People′s Hospital between August 1998 and December 2017 were performed. According to the inclusion criteria, a total of 56 patients with OEC were included in the study, including 13 patients concurrent with simultaneous endometrial lesions (Group A) and 43 patients with pure OEC (Group B).Results:Patients with synchronous endometrial lesions accounted for 23% (13/56). Mean age of Group A at diagnosis was (44.9±8.3) years old, 2/13 of patients were postmenopausal, and no one had a history of hypertension, the first symptom of 5/13 people was irregular vaginal bleeding. Mean age of Group B patients at diagnosis was (52.7±10.2) years old, 53% (23/43) of patients were postmenopausal, and 28% (12/43) patients had the history of hypertension, the first symptom of 4 (9%, 4/43) people was irregular vaginal bleeding. The differences of age, menopause status, history of hypertension and initial symptoms between the two groups were statistically significant (all P<0.05). There were no significant differences in fertility history, dysmenorrhea history, age of menarche, history of endometriosis, preoperative and postoperative CA 125 level, International Federation of Gynecology and Obstetrics (FIGO) stage, tumor grade, metastatic site and platinum-based chemotherapy drug resistance between the two groups (all P>0.05). The overall 5-year survival rate of OEC patients was 91.6%, and the overall 5-year progression-free survival rate was 76.6%. Among them, the 5-year survival rate of the OEC concurrent with simultaneous endometrial lesions group was 80.2%, and the pure OEC group was 93.4%; the 5-year progression-free survival rate of the OEC concurrent with simultaneous endometrial lesions group was 74.1%, and the 5-year progression-free survival rate of the pure OEC group was 77.3%. There were no significant differences between the two groups (all P>0.05). Multivariate analysis showed that the independent factors for the prognosis of OEC patients were FIGO stage ( P=0.006) and residual lesion size ( P=0.020). Conclusions:OEC patients have a high proportion of simultaneous endometrial lesions. OEC with simultaneous endometrial lesions are younger than patients with pure OEC. Synchronous endometrial lesions do not affect the prognosis of patients with OEC.

Objective To investigate the associated factors and trends of prehospital delay in elderly patients with acute ischemic stroke (AIS).Methods Elderly patients with AIS admitted to the First People's Hospital of Qujing from 2007 to 2017 were enrolled retrospectively.The data of patients was collected from the medical records.Onset-to-door time ＞ 2 h was defined as prehospital delay.The demographic and baseline data were compared between the delay group and the non-delay group.Multivariate logistic regression analysis was used to determine the associated factors for prehospital delay.In addition,the trends of prehospital delay time at the different stages of the study were also analyzed.Results A total of 1 566 patients with AIS aged ≥65 years were enrolled.Their mean age was 75.61 ±6.06 years.The mean time of prehospital delay was 10.83 ± 7.47 h (median time 8.27 h).Multivariatelogistic regression analysis showed that advanced age (odds ratio [OR] 1.271,95％ confidence interval [CI] 1.029-2.896;P =0.039),nocturnal onset (OR 1.413,95％ CI 1.067-3.859;P=0.013),and atypical symptom onset (OR 2.345,95％ CI 1.184-8.126;P=0.029) were independently positively correlated with prehospital delay,while the emergency medical service transport (OR 0.743,95％ CI 0.261-0.998;P =0.010),having medical insurance (OR 0.219,95％ CI 0.015-0.799;P =0.042),and having a bystander at the time of onset (OR 0.618,95％ CI 0.149-0.814;P=0.003) were independently negatively correlated with prehospital delay.At the different stages of the study,January 2007 to October 2010,November 2010 to April 2015,and May 2015 to December 2017,the mean time of prehospital delay was 12.59 ± 7.06 h,10.57 ±7.78 h,and 8.47 ±7.07 h,respectively.They showed a decrease trend,but the difference was not statistically significant.Conclusion Advanced age,nocturnal onset,and atypical symptom onset were the independent risk factors for prehospital delay,while emergency medical service transport,having medical insurance,and having a bystander at the time of onset were the independent protective factors for prehospital delay.The delay time of the elderly patients with AIS is declining year by year,but the improvement is not significant.The delay in seeking timely medical intervention remains an important public health problem.