BACKGROUND:To build up an effective method of isolating and culturing granule cels is a pivotal step to enhance fertilization-embryo transfer rate. Current studies mainly focus on the isolation methods of human ovarian granulosa cels rather than cel counting, purity and subsequent growth. OBJECTIVE: To establish the effective methods of isolating, purifying and culturing human ovarian granulosa cels in vitro. METHODS: Folicular fluid was harvested from women undergoing fertilization-embryo transfer procedures. Human ovarian granulosa cels were obtained from the folicular fluid by lysis treatment, precipitation method or density gradient centrifugation. Granulosa cel mucus masses were digested with type I colagen enzyme or hyaluronidase and then cultured in the culture medium with or without autologous folicular fluid. RESULTS AND CONCLUSION: Lysis treatment yielded the largest amount of granulosa cels compared to the precipitation method and density gradient centrifugation (P > 0.05,P < 0.05, respectively). Cels prepared by the three methods showed the same cel viability. After 24 hours of culture, the precipitation method obtained the largest amount of adherent granulosa cels (P < 0.05); and the density gradient centrifugation obtained the least amount of cels (P < 0.05). Compared with type I colagen enzyme, hyaluronidase took less time to digest the cels thoroughly. Autologous folicular fluid could promote the growth and survival of granulosa cels. These findings indicate that the precipitation method, though time-consuming, can obtain the highest cel viability and harvested the largest amount of granulosa cels after culture; hyaluronidase is more suitable for digesting granulosa cel mucus mass than type I colagen enzyme; autologous folicular fluid added into the culture medium is more conducive to granulosa cel growth.

Objective:To study the anti-inflammatory effect of the ethanol extract of Coptis chinensis Franch.in vitro.Methods: An inflammatory cell model was established by stimulating the mouse peritoneal macrophages in vitro with lipopolysaccharide(LPS).LPS stimulated of RAW264.7 cells for a long time after administration of intervention.Effect of ethanol extract of Coptis chinensis Franch.on RAW264.7 cell growth activity was analyzed by MTT assay.The production of tumor necrosis factor-α(TNF-α),IL-1β,IL-6,NO,prostaglandin E2(PGE2) was determined by ELISA.The expression of mRNA of TNF-α,induced nitric oxide synthase(iNos) and HO-1 was detected by real-time fluorescent quantitative PCR(RT-PCR).Results: The ethanol extract of Coptis chinensis had no inhibition effect on the scope of RAW 264.7 cells the scope of 5-80 mg/L.Each treatment group concentration of interleukin-6 (IL-6),interleukin-1β (IL-1β),TNF-α,NO,prostaglandin E2 (PGE2) content of LPS stimulation model group were significantly (P<0.01),and content was not related to concentration.Real-time quantitative (RT-PCR) showed,the concentration of each treatment group were significantly lower iNos,HO-1 and TNF-α mRNA expression (P<0.05,P<0.05,P<0.01),and content was not related to concentration.Conclusion: Coptis chinensis Franch.ethanol extract has anti-inflammatory effects in vitro,the mechanism may be related to inhibition of TNF-α,NO and other inflammatory factors and the impact of the activation of arachidonic acid (AA) metabolism.

Hematoma enlargement is a common harmful event after cerebral hemorrhage, which can lead to deterioration of neurological function and poor outcome. Early detection of high-risk patients can help prevent hematoma enlargement and improve the outcome of patients. Although the exact mechanism of hematoma enlargement is unclear, more and more evidence shows that many imaging signs may be related to hematoma enlargement, such as dot sign, mixed sign, black hole sign, island sign, etc. This article mainly reviews the relevant imaging prediction factors of hematoma enlargement from the perspective of imaging.

Objective:To investigate the anti-fatigue phenomenon induced by forced swimming in the mice,and to explore the anti-fatigue effect of argininyl fructoyl glucose (AFG) from red ginseng in the mice and its mechanism.Methods:The AFG was extracted from red ginseng.The ICR mice were divided into blank control group,low dose of AFG group (100 mg · kg-1),middle dose of AFG group (200 mg · kg-1) and high dose of AFG group (400 mg · kg-1) (n=20).The mice mere given a forced swimming test after continuous gavage for 28 d.The weights,organ indexes,time of forced swimming,contents of lactic acid (LD),blood urea nitrogen (BUN),hepatic glycogen (Gly) and expressing levels of PGC-1α in gastrocnemius of the mice in various groups were detected.Results:Compared with blank control group,the weights and organ indexes of the mice in low,middle and high doses of AFG groups had no significant differences (P＞ 0.05).Compared with blank control group,the time of forced swimming,contents of Gly and expressing levels of PGC-1α of the mice in low,middle and high doses of AFG groups were significantly increased (P＜0.05 or P＜0.01) in a dose-dependent manner.Compared with blank control group,The contents of LD and BUN in serum of the mice in low,middle and high doses of AFG groups were significantly decreased (P＜0.01).Conclusion:AFG has anti-fatigue effect in mice,and its mechanism may be related to energy metabolism.

Objective:To investigate the sleep status of preschool children with a coordination development disorder (DCD).Methods:Implementing stratified cluster sampling, 38, 900 questionnaires were distributed in 1000 classes of 200 kindergartens randomly selected in 11 prefecture-level cities of China′s Zhejiang Province. A total of 35, 464 valid responses were returned. On the basis of the responses, 2251 of the children were of (6.35%) were deemed DCD-positive, while the rest formed a DCD-negative group. The Children′s Family Social Environment and Sleep Health Questionnaire (1-6 years old) and the children′s sleep habits questionnaire were employed to investigate their sleep status.Results:①Regular sleeping time and duration, and falling asleep within 20 minutes were significantly less common in the DCD-positive group. In the negative group, shifting sleep place at night, finding it hard to sleep in strange environments, waking up ≥2 times a night, being awoken by others in the morning, difficulty in waking up and getting up in the morning, and bad mood after waking were significantly less frequent. ②In addition to delayed bedtime, the incidence of refusal to go to bed, fear of sleeping in the dark or alone, bedwetting, restless sleep, sleepwalking, talking in sleep, being awakened by nightmares, teeth grinding, loud snoring, apnea, dyspnea, crying and sweating after waking up at night, and insufficient sleep were all significantly higher in the DCD-positive group. ③The DCD-positive group also reported more sleeping in the daytime on weekends, more time on playing electronic products along with shorter sleep duration at night. There was, however, no significant difference between the two groups in sleep duration on school days and all day on weekends.Conclusion:Preschool children with a DCD have different sleep habits from those developing normally. They are more likely to suffer from sleep problems which affect their sleep duration.