Objective:To observe the interventative effect of Ziyin-Huatan Decoction by regulating exosomes on subcutaneous tumor of mice with gastric cancer. Methods:MGC-803 cells were randomly divided into exosome control group, low-dose group and high-dose group. The low-dose group and high-dose group were intervened with Ziyin-Huatan Decoction of 25 μg/ml and 100 μg/ml respectively. After 48 hours, the exosomes secreted by MGC-803 cells in each group were extracted. Twenty BALB/c male mice were randomly divided into exosome control group, low-dose Ziyin-Huatan Decoction group, high-dose Ziyin-Huatan Decoction group and blank control group, with 5 mice in each group. Except for the blank control group, the mice in the other groups were injected with exosomes extracted from the cells of the corresponding group through the orbit, 10 μg/time for each mouse, once every other day, a total of 15 times; the blank control group was injected with the same amount of PBS. Then SGC-7901 cells were inoculated into mice to establish a tumor model. The tumorigenic rate and body weight of mice were observed. The levels of CD31, VEGF and bFGF in tumor tissues were detected by immunohistochemistry. Results:Compared with the blank control group, the tumor weight [(170.00 ± 10.00) mg vs. (343.33 ± 20.82) mg] and the expression of CD31 (37.43 ± 0.55 vs. 63.30 ± 0.85), VEGF (11.37 ± 1.19 vs. 70.30 ± 0.72) and bFGF (43.77 ± 1.53 vs. 84.97 ± 1.86) in the high-dose Ziyin-Huatan Decoction group significantly decreased ( P<0.05). Compared with exosome control group, the expressions of CD31, VEGF and bFGF in low and high dose Ziyin-Huatan Decoction groups were decreased ( P<0.05). Conclusion:Ziyin-Huatan Decoction can significantly inhibit the growth of subcutaneous tumor of gastric cancer in mice by regulating exudation, which may be related to the inhibition of tumor angiogenesis.

OBJECTIVETo discuss the mechanism of gambogenic acid (GNA) in inducing the apoptosis of melanoma B16 cells.

METHODThe inhibitory effect of GNA on the proliferation of B16 cells was measured by the methyl thiazolyl tetrazolium (MTT) assay. The effect of GNA on B16 cells was detected by the Hoechst 33258 staining. The transmission electron microscopy was used to observe the ultra-structure changes of B16 cells. The changes in PI3K, p-PI3K, Akt, p-Akt, p-mTOR, PTEN proteins were detected by the Western blotting to discuss the molecular mechanism of GNA in inducing the apoptosis of B16 cells.

RESULTGNA showed a significant inhibitory effect in the growth and proliferation of melanoma B16 cells. The cell viability remarkably decreased with the increase of GNA concentration and the extension of the action time. The results of the Hoechst 33258 staining showed that cells processed with GNA demonstrated apparent apoptotic characteristics. Under the transmission electron microscope, B16 cells, after being treated with GNA, showed obvious morphological changes of apoptosis. The Western blot showed a time-dependent reduction in the p-PI3K and p-Akt protein expressions, with no change in p-PI3K and p-Akt protein expression quantities. The p-mTOR protein expression decreased with the extension of time, where as the PTEN protein expression showed a time-dependent increase.

CONCLUSIONGNA could inhibit the proliferation of melanoma B16 cells and induce their apoptosis within certain time and concentration ranges. Its mechanism in inducing the cell apoptosis may be related to PI3K/Akt/mTOR signaling pathways.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Melanoma ; metabolism ; pathology ; ultrastructure ; Mice ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism ; Terpenes ; pharmacology ; Xanthenes ; Xanthones ; pharmacology

Objective To observe the effect and security of lovasatin treating hyperlipidemia in children with steroid resistance nephrotic syndrome(SRNS).Methods Thirty-seven children with SRNS were administered lovasatin with normal liver function before lipid-lowing treatment.The changes of plasma lipids [total cholesterol(TC),trigly ceride(TG)] and lipoprotins [low density lipoprotein(LDL),very low density lipoprotein(VLDL),high density lipoprotein(HDL)],albumin(Alb),serum creatinine(Scr),aminotransferase(ALT),24 hours of urinary protein and drug side-effects were observated for 4 weeks.All children treated with regular glucocorticoids therapy for 2 months still presented with urinary protein significant positive.Results There were decreasing of plasma lipids and lipoproteins after 2 weeks of lovasatin treatment,especially for TG,24 hours of urinary protein(Pa

AIM: To investigate the effect of Chinese propolis on the activity of phosphatidylcholine-specific phospholipase C (PC-PLC) and the expression of Toll-like receptor 4 (TLR4) in LPS-treated vascular endothelial cells (VECs). METHODS: Confluent VECs were stimulated with LPS at the concentration of 100 μg/L in the presence of 0.5% fetal bovine serum. The cells were treated with Chinese propolis at the concentration of 12.5 mg/L for 12 h and 24 h. The viability of VECs and the level of nitric oxide (NO) were detected by sulforhodamine B (SRB) assay and chemical method, respectively. The activity of PC-PLC was measured using L-α-phosphatidylcholine as substrate. The protein levels of TLR4, nuclear factor-Κb p65 (NF-Κb p65) and p53 were determined by Western blotting. The level of intracellular reactive oxygen species (ROS) was examined using a fluorescent probe, 2',7'-dichlorodihydrofluorescin (DCHF). For the measurement of mitochondrial membrane potential, the fluorescent dye JC-1 was used. RESULTS: Treatment with Chinese propolis for 24 h had no effect on the viability of VECs. However, the levels of NO and ROS were significantly decreased by Chinese propolis. PC-PLC activity and NF-Κb p65 expression were significantly depressed by Chinese propolis treated for 12 h, and the expression of TLR4 and p53 were dramatically decreased by Chinese propolis treated for 12 and 24 h. No effect of Chinese propolis on mitochondrial membrane potential was observed. CONCLUSION: Chinese propolis depresses the activity of PC-PLC and the expression of TLR4, and then inhibits the downstream signal molecules such as NF-Κb p65, p53, ROS and NO in VECs.

Objective To evaluate the significance of standardized ultrasound examination of fetal structures in the ifrst trimester (11-13+6 weeks). Methods The ultrasound data of 29 858 fetuses who underwent the ifrst trimester screening were analyzed retrospectively in Beijing Obstetrics and Gynecology Hospital of Capital Medical University from January 2010 to December 2012 and followed up the cases with structural abnormalities and increased nuchal translucency (NT). Results Among 29 858 fetuses, 328 structural defects were detected in 284 fetuses (0.95%) by ultrasound in the first trimester, including fetal hydrops (124 cases), choroid plexus cysts (36 cases), exencephaly (32 cases), gastroschisis (24 cases), exomphalos (22 cases), cardiac defects (19 cases), megacystis (14 cases), spine abnormalities (10 cases), meningoceles/encephaloceles (9 cases), alobar holoprosencephaly (8 cases), hydrocephaly (7 cases), abnormalities of extremities (7 cases), acrania (6 cases), amniotic band syndrome (4 cases), abdominal cyst (3 cases) and conjoined twins (3 cases). Pregnancy was terminated in all cases (115) with exencephaly, gastroschisis, exomphalos, meningoceles/encephaloceles, alobar holoprosencephaly, hydrocephaly,abnormalities of extremities, acrania, amniotic band syndrome and conjoined twins, and the defects were verified by the gross appearance of fetuses. Two cases of megacystis were turned out to be normal during follow-up while 1 case was proved to be aneuploid and 1 other case progressed to multiple abnormalities during the second trimester. Most choroid plexus cysts, except 1 case of aneuploid, were normal during the second and the third trimester follow-ups by ultrasound. Thirteen cases of fetal hydrops were found to be aneuploids and 4 hydrops cases were proved to be complicated with other structural defects during the second and the third trimester. NT was increased in 422 cases (1.41%), among which 122 cases (28.91%) were complicated with structural defects and/or fetal hydrops. Ninty-nine cases with increased NT underwent chromosomal examination and 38 cases were found to be aneuploides. During the follow-up of 298 cases with increased NT who continued pregnancy, 21 structural abnormalities were found, including cardiac defects (14 cases), neural abnormalities (4 cases) and diaphragmatic hernia (3 cases). Conclusion The standardized fetal ultrasound in the first trimester is a effective tool for the screening of aneupolides and major structural abnormalities.

ObjectiveTo study the effects on growth characteristics of cells mutated by outer space. Methods5 tumor cell lines and 1 bio-pharmaceutical cell line were carried in the recoverable satellite No.18 and returned after 18 days flying in outer space.The cell living systems were utilized in passive carrying. Some survival cells were monocoloned, and the experimental methods such as cell morphological observation,MTT assay and FACS were used for analysis of cell growth characteristics. ResultsThe survival cell rates of tumor cell lines and bio-pharmaceutical cells were markedly different. Compared with the control group, mutated cells appeared to have multiple cell morphological changes and the cell number of G1 phase was increased markedly (P<0.05). Growth rate in mutated cells were varied without specific discipline. ConclusionEffects of outer space on growth characteristics were complicated and multiplex in mutated cells,which were not all heredity in further passage. The differences of the monoclonal cell lines provide the potential to obtain optimizing cell lines by screen.

ObjectiveTo develop the facility of mammalian cell culture in hypothermia for long-term, and to lay the base for carrying mammalian cells to outer space and establishing outer space biomedical research stage. MethodsMultiple cell lines were maintained in the spacial designed hypothermia cell culture facility and carried to space by Shenzhou 4 and No.18 recoverable satellite. Cell morphology, cell multiplication and the ability to be monocloned were observed.ResultsCells were successfully maintained in the hypothermia cell culture facility for up to 26 days without obvious changes of cell morphology. The cells could normally grow, multiple and be monocloned after inoculated in 37 ℃ for 8 h.ConclusionA hypothermia cell culture facility was successfully established,which ensure technically mammalian cells to be carried by space craft and further research on space radiation.

To establish HPLC specific chromatogram and its correlation with the protection effect of Shuanghuanglian on MDCK (Madin-Darby canine kidney) cell injury induced by influenza A virus( H1N1). Nine recipes of Shuanghuanglian based on the official prescription were prepared according to orthogonal test for HPLC analysis and MDCK cells protection experiment separately (cytopathic effect (CPE) method was used for observing the virus infectivity and MTT staining results were used as the determining indexes for drug concentration selection and analyzing cell viability). The results suggested that all the other Shuang-Huang-Lian recipes except recipe1 demonstrate protecting effect on MDCK cell injury induced by influenza A virus (P < 0.01, P < 0.001). Stepwise regression analysis was used for analyzing the relationships between HPLC fingerprint and the protecting effect of Shuanghuanglian on influenza A virus induced MDCK cell injury. Peak 2, 3, 6, 8 and 12 were found to be strongly related with anti-influenza A virus efficacy. Stepwise regression analysis of recipes data and efficacy data showed that Lonicerae Japonicae Flos and Forsythiae Fructus were positively associated with the protecting effect of cells injury. From HPLC fingerprints, we found that peak 2, 3, 12 were from Lonicerae Japonicae Flos and peak 6, 8 were from Forsythiae Fructus. Four peaks were identified through comparing the retention time between the standard and Shuanghuanglian recipes, and they were chlorogenicacid, cryptochlorogenic acid, forsythoside B and 3,4-dicaffeoylquinic acid respectively. Caffeic acid derivatives in Lonicerae Japonicae Flos and Forsythiae Fructus were found to be greatly correlated with anti-influenza A virus efficacy and maybe the substance basis of Shuanghuanglian.
Animals ; Antiviral Agents ; analysis ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Dogs ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Forsythia ; chemistry ; Influenza A Virus, H1N1 Subtype ; drug effects ; physiology ; Lonicera ; chemistry ; Madin Darby Canine Kidney Cells ; Scutellaria baicalensis ; chemistry

Objective To investigate the tendency of distribution and drug-resistance of the causative organisms of urinary tract infections(UTIs)in Wuhan,and provide reliable evidence for clinical treatment.Methods Analyzed the 5 378 stains of pathogen isolated from the urine of patients in hospital.The bacteria isolates were identified with BD Phoenix-100 while can-dida isolates were identified by color plate.Results A total of 5 378 stains of pathogen had been isolated.There were 2 945 stains (54.8%)of Gram-negative bacteria,1 657 stains (30.8%)of Gram-positive bacteria,776 stains (14.4%)of fungus. The rates of Escherichiacoli resistant to penicillin were highest (>83%),and there were no carbapenem-resistant strains. There were vancomycin and linezolid-resistant Enterococcispp strains,the lowest dection rates of which were 0.3%.The de-tection rate of MRCNS was over 83%.Conclusion Escherichiacoli was the most common pathogens of urinary tract infec-tion,and theβ-lactamase inhibitor complex can be used as empirical treatment of E.coli infections.Thedetection rate of MRCNS increased,which shoud be kept a watchful eye on.

Objective To study the effects of curcumin mediated photodynamic therapy (PDT)on the growth and proliferation in human cervical cancer cell line H8 in vitro and in vivo,and to investigate its antitumor mechanisms.Methods The effects of curcumin mediated PDT on proliferation of human cervical cancer H8 cell by MTT assay was used to screen the optimal parameter.Changes in cell morphology were observed by May-Gr ünwald-Farbstoff Giemsa staining.The apoptosis rate was estimated by flow cytometry.The effect of PDT by curcumin on the expressions of Bcl-2,P53 and survivin in H8 cells was detected by fluorescence real-time reverse transcription-polymerase chain reaction (RT-PCR).Forty BALB/C nude mice underwent subcutaneous injection of H8 cell line so as to establish animal models,and then were randomly divided into four equal groups:control group,irradiation alone group,curcumin alone group,curcumin PDT group.HE staining and pathological examination were performed.Immunohistochemical study was conducted to detect the protein expression of the apoptosis inhibiting genes of Bcl-2.Results The proliferation inhibition of H8 cells was obvious after PDT when curcumin 5μmol/L with irradiation 100 J/cm2,and with dose dependent manner.Typical morphologic features of apoptosis appeared characterizedly by marked chromatin condensation,nuclear pyknosis and fragmentation,and the appearance of apoptotic bodies.The total apoptosis rate was higher in PDT group [(47.21 ± 4.11)％]than in control group(1.71 ±0.16) ％ (P＜0.01).The mRNA expression of Bcl-2,P53 and survivin in H8 cells were suppressed significantly.HE staining showed remarkable subcutaneous necrosis in the PDT group.Immunohistochemistry showed remarkable down-regulation of protein expression of Bcl-2(P＜0.01).Conclusions Curcumin-mediated photodynamic therapy has a significant killing effect on H8 cells in vivo and in vitro.Its antitumor effect might be related to induction of Tumor cell apoptosis and suppression of Bcl-2 mRNA and protein expression.