Objective: To seek the optimum technology of drying and to solve the problem of powder sticking clump. Methods : On the basis of major drying condition, such as duration of drying, temperature of drying, evaluating different drying technology; On the basis of the quantity of the powder sticking on the wall and the hygroscopicity of one, investigating different excipients how to affect the spray drying. Seeking the optimum kinds and quantity of excipients. Results : The best properties of the granules are from the spray drying process. ?-cyclodextrin and lactose have the effects on solving the powder sticking on the wall. Conclusion : The spray drying process is the optimum one. Adding 3～5 percent of ?-cyclodextrin has an effect on improving the phenomenon of powder sticking on the wall in process of the spray drying.

Objective To investigate the specific anti-tumor immune response induced by dendritic cells (DCs) transfected with total RNA of human pancreatic cancer Capan-2 cells.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) derived from six patients with pancreatic cancer.Total RNA of Capan-2 cells and MUC4 mRNA were transfected into DCs by electroporation.The survival rate of transfected DCs was determined by MTT method and the expression of MUC4 mRNA in DCs was detected by Western blotting.The activity of cytotoxic T lymphocyte cells (CTLs) induced by DCs transfected with total RNA of Capan-2 cells were evaluated by IFN-γ ELISA and the induction of specific CTL response to the killing effect on pancreatic cancer cell in vitro were measured by 51 Cr standard cytotoxicity test.Results The survival rate of DCs transfected with total RNA of Capan-2 cells (DC-Capan-2-total RNA) showed a decrease in a time dependent manner and the survival rate was reduced to 60.81％ after transfection for 96 h.The survival rate of MUC4 mRNA transfection DCs (DC-MUC4 mRNA) was stable at around 80％.The difference of DCs surviral rate between the two groups was statistically significant (P ＜ 0.05).The amount of MUC4 protein expression of DC-Capan-2-total RNA was significantly lower than that of DC-MUC4 mRNA (P ＜0.05).The quantity of CTL IFN-γ release induced by DC-Capan-2-total RNA was (89.34 ± 3.85)U/mL and the quantity of DC-MUC4 mRNA induced CTL IFN-γ release was (21.77 ± 21.77)U/ml There was statistically significant between the two groups (P ＜0.05).In addition,the specific CTLs induced by DC-Capan-2-total RNA could effectively identify and kill the HLA-A2+/MUC4+ Capan-2 and the HLA-A2+/MUC4-PANC 1 cells,and could not effectively identify and kill the HLA-A2 /MUC4-MiaPaCa-2 cells and the HLA-A2-/MUC4 + AsPC-1 cells.Conclusions A more pronounced CTL anti-tumor immune response can be induced by DCs transfected with total RNA of Capan-2 cells compared with a single tumor associated antigen,but it is limited by MHC class Ⅰ antigen presented.

Adult ; China ; Female ; Humans ; Occupational Health Services ; Women's Health Services ; Young Adult

Objective To study the effect and mechanism of chlorogenic acid in allergic rhinitis(AR) mice.Methods The model of AR mice was induced by intraperitoneal injection of adjuvant suspension on every other day for 7 times, and then by nasal drip of 10% OVA for 10 days.After administration with 50, 100, 200 mg/kg of chlorogenic acid for 30 min, AR mice were attacked locally for 10 days.Behavior analysis were observed and the effect of chlorogenic acid on the contents of cytokines, IgE and histamine by ELISA in AR mice were determined.Results Chlorogenic acid decreased the number of nose-scratching events in 10min, the weight of nosal draingage, the index of spleen, the content of IL-4 in nasal lavage fluid ( NLF) and the contents of cytokines, IgE and histamine in serum; while increased the index of thymus, the content of IFN-γin NLF, the contents of IFN-γin serum and the rate of IFN-γ/IL-4.The data showed a statistically significant up-regulation of IFN-γmRNA expressions and down-regulation of IL-4 mRNA, IL-5 mRNA, IL-10 mRNA expressions after adiminstration of chlorogenic acid.Conclusion Chlorogenic acid might exert the effect in AR mice through regulating the balance of Th1 and Th2.

Objective To investigate the clinical features of macrophage activation syndrome (MAS) associated with systemic lupus erythematosus (SLE).Method The clinical data of 15 patients with SLE-induced MAS diagnosed in Peking Union Medical College Hospital from July 2011 to December 2014 were retrospectively analyzed.Results Fourteen patients were female.The average age was 28.07.When MAS occurred,the average duration of SLE was 20.47 months,and the average SLE disease activity index (SLEDAI) was 18.4.All 15 patients developed fever,hematocytopenia and impaired liver function in the course of MAS,while patients with splenomegaly,coagulation disorders and neuropsychiatric symptoms were 11,14 and 8,respectively.All 15 patients presented leukocpenia and thrombocytopenia.Hypofibrinogenemia,elevated ferritin and hemophagocytosis in bone marrow were respectively observed in 7,11 and 12 patients.Glucocorticoids were used in all patients,among whom eight received pulse methylprednisolone therapy.Thirteen patients were treated with immunosuppressants,including cyclosporine A,tacrolimus,cyclophosphamide and mycophemolate mofetil.Complete remission was achieved in 14 patients.One patient died of MAS.Conclusions In patients with SLE,MAS was most commonly seen in young females with short SLE duration and active disease.Fever,splenomegaly,hematocytopenia,coagulation disorders and liver damage are the most remarkable clinical manifestations.Early diagnosis and intensive therapy are the key parts to improve clinical outcome.

Objective To study the effects of Weibimei and its Chinese medicine portions(cortex frangulae, fennel powder, acorus tatarinowii, glycyrrhizae and asparagus) on cell cycle regulation in gastric cancer cells and growth inhibition in gastric cancer Xenografts.Methods Gastric cancer SGC-7901 cells were normally cultured and treated with Weibimei, extractum glycyrrhizae, cortex frangulae, fennel powder, acorus tatarinowii and asparagus.Then, the proliferation of SGC-7901 cells were detected by MTT assay, their effect on SGC-7901 cell cycle were measured by flow cytometry, and the expression of cell cycle related proteins were examined by Western blot.Animal models of SGC-7901 cells xenografts in nude mice were constructed to evaluate the growth inhibition effects of Weibimei and its Chinese medicine portions, and the expression of STAT3,p-STAT3, Cyclin D1, Bcl-2 and Survivin in gastric tumors were detected by immunohistochemical assays.Results Weibimei and its three kinds of Chinese medicine portions, cortex frangulae, fennel and acorus tatarinowii could obviously inhibit the proliferation of SGC-7901 cells, compared to control group, with a statistical significant difference (P<0.05).The cell cycle of SGC-7901 was arrested at G1 phase with cortex frangulae, fennel and acorus tatarinowii treatment, the expression of Cyclin A, B, D, E were decreased significantly (P<0.05).However, no significant effects was found in glycyrrhizae or asparagus group.Xenografts tumor sizes in Weibimei, cortex frangulae, fennel or acorus tatarinowi groups were much smaller than that in saline group or Triple Therapy group (three-kinds-drugs-combination for the treatment of gastric ulcer) (P<0.05).Xenografts experiments showed that the tumor growth in nude mice in Weibimei, cortex frangulae, fennel or acorus tatarinowi groups were significantly smaller than that in control group or in Triple Therapy Group (P<0.05), and phosphorylated STAT3, and STAT3 signaling targeted genes, including Bcl-2, Cyclin D1 and Survivin were all significantly down-regulated by immunostaining.Conclusion Weibimei and its Chinese medicine portions including cortex frangulae, fennel and acorus tatarinowii can could significantly inhibit the growth of SGC-7901 cells proliferation and arrested the cell cycle of G1 phase in vitro, and inhibit gastric cancer xenografts in nude mice in vivo.

Objective To investigate the biological effects of Wnt gene in kidney cancer Caki?2 cells. Methods The Wnt gene was silenced in kidney cancer Caki?2 cells by lentivirus vector. The cell proliferate ability of cells in each group were assayed by CCK?8 kit at different time points. The apoptosis of Caki?2 cells was observed after silencing Wnt gene by transmission electron microscope. The invasion ability of each group cells was tested using Transwell chambers. The genes expression changes of Wnt/β?catenin signaling pathway and apoptosis related gene were determined by realtime PCR. Results Compared with the other two groups,the cell proliferate ability of the cells after silencing Wntgene was suppressed,and the difference was statistically significant(P<0.05). Apoptosis increased significantly in shRNA+Caki?2+Wnt group cells with silencing of Wntgene, and apoptotic body appeared in these cells. In invasive experiment,the number of emigrated cells in shRNA+Caki?2+Wnt group were significantly lower than other groups(P<0.05). The expression of Wnt mRNA,β?catenin mRNA and Bcl?2 mRNA in shRNA+Caki?2+Wnt group cells was lower than other groups(P<0.05). Conclusion Silencing of Wnt gene of kidney cancer Caki?2 cells can affect the proliferation rate of the cells, promote the cell apoptosis,and inhibit the invasion ability,which provide certain theoretical basis for the research and development of new drugs and new therapeutic targets.

AIM: To study the relation between prolon ge d survival of skin allograft by chuan-ke-zhi (CKZ, drug of Chinese herbal) and C D4+CD25+ regulatory T cells (CD4+CD25+ Tr) in mice. METHODS: Skin allograft and isograft model in mice were establi shed and CKZ was administered by intraperitoneal injection. To observe its influ ence on survival of the graft, three color fluorescent staining together with fl ow cytometry was used to analyze the change of CD4+CD25+ Tr. RESULTS: The survival of skin allograft in CKZ group was signifi cantly prolonged compared to control group, (19.5?2.3) days and (10.2?2.2) days, respectively, P0.05). CONCLUSION: CKZ has an effect of prolonging the survival of skin allograft. Enhancement of CD4+CD25+ Tr might be one of the mechanisms under lying its immunosuppressive effect.

ObjectiveTo study the teasibility ot using the recipient cystic duct for biliary reconstruction in living donor liver transplantation.MethodsBetween January 2008 and September 2011,5 patients underwent living donor liver transplantation with biliary reconstruction.The recipient 's cystic duct was anastomosed with one branch of the donor's bile duct,and other donor's bile ducts were anastomosed with hepatic duct or right hepatic duct.After transplantation,the recipients were followed up with liver function test.Radioscopy was preceded at 2nd week and 3rd month after transplantation.ResultsTwo weeks after transplantation,cholangiography through the tubes showed no leakage or stricture at intrahepatic and extrahepatic bile ducts. Three patients recovered well without any hepatic dysfunction,and cholangiography showed well at 3rd month after operation.One patient had normal hepatic function after operation,and had not yet taken cholangiography.One patient recovered smoothly early,but after 3 months the cystic duct was tortuous with anastomotic stricture.The liver function was normal,and T tube was withdrawn.But 9 months later,hepatic dysfunction appeared,and the middle of the lumen was slightly narrowed in MRCP.Tacrolimus was replaced by Sirolimus,and the liver function returned to normal.ConclusionBiliary reconstruction using the cystic duct is feasible and safe for living donor liver transplantation.

Objective To investigate the anti-tumor immune response induced by human pancreatic cancer mucin 4 mRNA and human telomerase reverse transcriptase (hTERT) mRNA cotransfected dendritic cells (DC),and to provide the experimental evidences for the treatment of pancreatic cancer with multi-epitope loaded DC vaccine.Methods DC were isolated from peripheral blood mononuclear cells (PBMC) of six patients with HLA-A2+ pancreatic cancer and cultured.Mucin 4 mRNA and hTERT mRNA were transcripted and amplified in vitro,which were transfected into DC separately or in order by eleetroporation.DC were cultured for 48 hours.The expressions of mucin 4 and hTERT in DC were detected by quantitative real-time polymerase chain reaction (PCR) and Western blot.The survival rates of transfected DC were determined by methylthiazolyl tetrazolium (MTT) method.The cytotoxic T lymphocyte (CTL) activation induced by mucin 4 mRNA and hTERT mRNA transfected DC were evaluated by interferon (IFN)-γ release assays (enzyme linked immunosorbent assay) method.The cytotoxicity of CTL induced by mucin 4 mRNA and hTERT mRNA transfected DC in pancreatic cancer cell lines MiaPaCa-2,Capan-2,AsPC-1 and Pane-1 was measured by 51Cr standard cytotoxicity test.Student t test was performed for statistical analysis.Results After in order transfection of mucin 4 mRNA and hTERT mRNA for 48 h,the relative quantity of the expression of mucin 4 and hTERT in DC were 30.09±5.24 and 12.87±3.36,which were lower than the relative quantity of the expression in DC transfected separately (38.54±6.21 and 36.35±5.03,t=3.469,6.721,both P＜0.05).After transfected in order for 96 hours,the survival rate of DC decreased to 52.17％,which was lower than that of DC transfected separately (around 80％).The quantity of IFN-γ releasing of specific CTL induced by mucin 4 mRNA and hTERT mRNA cotransfected DC was (32.57±2.01) U/mL in 24 hours,which was higher than that of CTL induced by DC transfected with mucin 4 mRNA ((23.06±4.74) U/mL) or hTERT mRNA ((16.82±3.67) U/mL) separately (1=5.092,7.141,both P＜0.05).After co-transfected with mucin4 mRNA and hTERT mRNA,DC could effectivly induce HLA-A2+/mucin 4+/hTERT+ specific CTL immune responses,however there was no significant cytotoxicity in HLA-A2+ pancreatic cancer cells.Conclusion The induction of CTL anti-tumor immune response by DC co-transfected with mucin4 mRNA and hTERT mRNA is more significant compared with that by single antigen loaded DC.