Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.

OBJECTIVE:To establish an analytical method for the determination of prednisolone and prednisolone-hemisuccinate in biological media METHODS:Using RP-HPLC,biological media were extracted with a mixture of n-pentane and methyl tert-butyl ether(2∶3) The mobile phase consisted of acetonitrile and trisodium citrate buffer (33∶67) The sample was eluted on C18 column and detected at 254nm RESULTS:The extraction recoveries of two drugs were over 80% and the RSDs of within-day and day-to-day were below 3% CONCLUSION:The method is simple,rapid,sensitive,accurate and good enough to be applied to the pharmacokinetic study of new preparation of prednisolone and can also be applied to other similar glucocorticoids

OBJECTIVE:To verify the effect of Weichangkang capsule(WCK) on the function of gastrointestinal tract.MET-HODS:The effects on movement of charcoal powder in mice and secretion of digestive enzyme and absorption of D-xylopyranose into serum in rats were studied.RESULTS:WCK capsule markedly increased the movement of charcoal powder in mice,secretion of digestive enzyme and absorption of D-xylopyranose into serum in rats.CONCLUSION:WCK capsule could obviously improve the dysfunction of gastrointestinal tract.

Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

Objective:To explore the curative effect and toxic and side effects of Xihuang Capsule in the patients with advanced esophageal cancer treated with chemoradiotherapy, and to clarify its mechanism.Methods:The patients with advanced esophagealcancer without treatment were selected.The patients treated with Xihuang Capsule combined with concurrent chemoradiotherapy were defined as combination treatment group(n=27), and then other 27 patients were randomly selected from 87 patients treated with concurrent chemoradiotherapy alone as chemoradiotherapy group.The observation time was 90 d from the begining of chemotherapy.The differences in KPS, degree of dysphagia, incidence, occurrence time and degrees of acute radiation esophagitis of the patients in two groups were compared, and local control of tumor was analyzed.Results: After treatment, the KPS scores of the pateints in two groups were decreased, and the KPS score in combined treatment group was significantly higher than that in chemoradiotherapy group (P<0.05).The grade of dysphagia of the patients in combined treatment group was lower than that in chemoradiotherapy group during the late period of treatment(P<0.05).The incidence of acute radiation esophagitis, the occurence time of acute radiation esophagitis of the patients in combination group was lower than that in chemoradiotherapy group (P<0.05);the occurrence time of acute radiation esophagitis was later than that in chemoradiotherapy group (P<0.05);but there were no significant differences in the grades of acute radiation esophagitis and tumor local control between two groups(P>0.05).Conclusion:Xihuang Capsule can improve the quality of life, reduce dysphagia degree and the incidence of acute radiation esophagitis,and delay the occurrence time;it can be used as an effective auxiliary treatment of advanced esophageal cancer.

Objective To study the characteristics of immune function,Epstein-Barr virus(EBV) antibodies and EBV-DNA in children with different clinical types of EBV infection,which provide basis for prevention and treatment of EBV infection.Methods Clinical data of 103 patients suffering from EBV infection were retrospectively analyzed in Xijing Hospital of the Fourth Military Medical University.A total of 103 children were divided into infectious mononualeosis(IM) group(n=68),chronic active Epstein-Barr virus infection(CAEBV) group(n=13) and Epstein-Barr virus-related hemophagocytic lymphohistiocytosis(EBV-HLH) group(n=22).The changes of EBV antibodies,EBV-DNA,immunoglobulin levels,lymphocyte subpopulation and complement series were detected and compared among the three groups.A total of 26 healthy children at the same stage were enrolled as a control group,immunoglobulin levels,lymphocyte subpopulation and complement series were detected in control group,then compared with the rest of the three groups.Results The levels of C3 and C4 in CAEBV group and EBV-HLH group were significantly decreased than those in the control group(P<0.05).The levels of IgA,IgG and IgE in EBV-HLH group,CAEBV group,IM group and control group gradually increased(P<0.05,respectively).The levels of IgA,IgG and IgE in EBV-HLH group and CAEBV group significantly decreased than those in control group(P<0.05,respectively).The levels of IgA,IgG and IgE in IM group decreased than those in control group,while there were no statistically significant differences(P>0.05).CD8+T cells in IM group significantly increased than those in the rest of the three groups(P<0.05,respectively).T cells,CD8+T cells,CD4+ T cells,CD4+/CD8+ ratios,NK cells,B cells of EBV-HLH group significantly decreased than those in the rest of the three groups(P<0.05).The positive rates of EBV antibodies in CAEBV group and IM group were significantly higher than those in EBV-HLH group(P<0.05).EBV-DNA in EBV-HLH group were significantly higher than those of CAEBV group and IM group(P<0.05).Conclusion EBV-DNA levels in the serum are positively correlated with disease types and severity,the pathogenesis of IM,CAEBV and EBV-HLH induced by EBV infection are associated with immune dysfunction.Dynamic monitoring of EBV load and cell immune function can reflect disease status and progress risk.

OBJECTIVE:To determine the content of ?-schizandrin,one of the effective ingredients,in Shuangjia Wuling capsules METHODS:The RP-HPLC method was performed with YWG C18 column(4 6mm?250mm) The mobile phase consisted of methanol-water(72∶28) The detecting wavelength was 254nm RESULTS:The calibration curve for ?-schizandrin was linear in the range of 0 0 207～0 4 130mg/ml(r=0 9 999) The average recovery was 97 68% with RSD=1 85% CONCLUSION:The method is simple,rapid and reliable for quality control of the capsules

Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.

AIM: To investigate the contraction effect of essential oil of Radix Angelicae Sinensis on isolated uterus in rats. METHODS: Isolated uterine strips of rats were used to observe the reactions in the presence of oxytocine and high k + depolarization solution. RESULTS: The isolated rat uterus contraction induced by oxytocin and Ca 2+ after high K + depolarization was dose dependently inhibited by the essential oil. The cumulative dose response curves of CaCl 2 shifted to right and the maximal responses were depressed. This showed that the essential oil was a non competitive antagonist. The extracellular and intracellular Ca 2+ dependent contraction evoked by oxytocin could also be inhibited by the essential oil. CONCLUSION: The essential oil of Radix Angelicae Sinensis could inhibit the contractility of the isolated uteri, and it might act as an Ca 2+ antagonist to relax the isolated rat uterus.