AIM: To investigate the expression of Fas/FasL system in human lung carcinoma cell lines (A549, EBC-1, LCSC) and T-lymphocytes (Jurkat), and to search the possibilities of immune escape and counterattack mediated by Fas/FasL pathway in human lung carcinoma cells. METHODS: The protein and mRNA expression of Fas and FasL were detected by FACScan, RT-PCR, respectively. Cell apoptosis was assessed by fluorescent staining. Cell growth was determined by a trypan blue exclusion assay. RESULTS: Fas and FasL were expressed in 3 human lung carcinoma cell lines and T-cells, Jurkat. The lung carcinoma cells inhibited the growth (P

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.

ObjectiveTo validate the feasibility of the outer space carrying system in culturing the CHO(dhfr-)cells in hypothermia, and to observe the effects of this carrying system on the growth characteristics of the CHO(dhfr-) cells.MethodsThe growth characteristics of the CHO(dhfr-) cells were observed and analyzed with cell morphological observation, MTT assay, FCM,3H incorporation and chromosome after the cells were cultured for 25 days in the carrying system. ResultsComparing with the control group, the CHO(dhfr-) cells appeared multiple cell morphological changes, the difference of cell cycle was not significant, the broken chromosome was not seen, the cell growth speed decreased markedly and big molecular biosynthesis increased obviously. ConclusionThe outer space carrying system has no outstanding effects on the survival and heredity of the CHO(dhfr-) cells, so that it can be used in cell carry.

BACKGROUND:Immunity of fetal liver stem cel transplantation is rarely reported, syngeneic and al ogeneic fetal liver stem cel transplantation in the treatment of hepatic cirrhosis is stil unclear.
OBJECTIVE:To observe the therapeutic effects of syngeneic and al ogeneic fetal liver stem cel transplantation on hepatic cirrhosis as wel as immune rejections during the therapeutic process.
METHODS:The fetal liver stem/progenitor cel s from BALB/c and C57BL/6 mice were isolated and purified by the type IV col agen enzyme digestion method. A total of 104 healthy BALB/c mice were randomly assigned to four groups. Normal control group:no treatment;Hepatic cirrhosis group, syngeneic transplantation group and al ogeneic transplantation group:16 weeks after hepatic cirrhosis models of mice were developed by intraperitoneal injection with carbon tetrachloride, physiological saline, syngeneic fetal liver stem cel s and al ogeneic fetal liver stem cel s were injected via the caudal vein. Final y, the survival statuses, liver function, hepatic fibrosis index, the number and ratio of immune cel s (CD4+T, CD8+T, NK, NKT) and histopathologic examinations were compared in each group after transplantation 4 weeks.
RESULTS AND CONCLUSION:The survival rates in the two transplantation groups were both 100%, which was significantly higher than that in the hepatic cirrhosis group (67%, P<0.05). The liver function and liver fibrosis index in each group did not show statistical differences (P>0.05). Immunological tests showed no difference between groups (P>0.05). Pathohistology examination of hepatic tissue repair:Al ogeneic transplantation group>syngeneic transplantation group>hepatic cirrhosis group. Hence, fetal liver stem cel transplantation via the caudal vein could elevate the survival rate of hepatic cirrhosis mice, al eviate the degree of hepatocyte necrosis. There is no immunologic rejection during syngeneic and al ogeneic fetal liver stem cel transplantation that could help to treat hepatic cirrhosis in mice.

Objective To establish a stable transplantation tolerance model by combined treatment with PTD-mFoxp3 fusion protein and allogeneic bone marrow transplantation and study its application to reduce the incidence of graft-versus-host disease (GVHD).Method BALB/c mice as recipients were randomly divided into four groups,accepted medical linear accelerator ray 3.0-Gy total body irradiation (TBI) before bone marrow transplantation (BMT),and injected with donor C57BL/6 mice bone marrow cells 3 × 107withinn 4-6 h.The BALB/c mice in group A were given PTD-mFoxp3 fusion protein through tail vein intermittently (day-2,0,1,3,5,7,9,11,13),those in group B given the same dose mFoxp3 protein,those in group C given normal saline,and those in blank control group given no treatment.The symptoms of GVHD were observed after BMT.Chimerisms were determined on the week 1,2,4,8 and12 post-BMT by flow cytometry.Liver and intestinal tissues were collected for pathological examination.Recipient immune response was assessed on the week 4 and 12 by mixed lymphocyte reactions (MLR) after BMT.Results The chimerism level in group A was high [(38.16 ± 3.09) ％] in the first 12 weeks after BMT,and that in group B and group C was reduced [(20.12 ± 4.75) ％ and (15.72 ± 2.36) ％ respectively,P＜0.05].MLR revealed that shown lymphocyte donor-derived lymphocyte proliferation rate at 4th and 12th week in group A was significantly lower than in other groups (P＜0.05).Pathological examination showed infiltration of lymphocytes in the liver and intestine was milder in group A than in other groups.Conclusion PTD-mFoxp3 fusion protein combined with allogeneic bone marrow transplantation can effectively establish a stable transplantation chimeric model and reduce the incidence of GVHD.

Diagnostic Errors ; Encephalocele ; diagnosis ; Female ; Humans ; Meningocele ; diagnosis ; Middle Aged ; Mucocele ; diagnosis ; Sphenoid Sinus ; pathology

Objective To investigate the effects of all-trans retinoic acid (ATRA)-intragastric-administration on the survival time of mouse skin allografts and the role of interleukin (IL)-23 and IL-17 thereof. Methods The skin trans-plantation of mice was done by DBA/2 as donors and Balb/c as recipients. The recipients were divided randomly into three groups:control group, low-dose group and high-dose group. Mice of the corresponding groups were intragastrically adminis-tered corn oil, 10 mg/kg ATRA and 30 mg/kg ATRA respectively from 1 day before the transplantation to the 14th day after the transplantation. The survival time of transplanted skin was observed after the operations. Skin grafts of mice were harvested for histopathological examination in three groups. The serum levels of IL-23 and IL-17 were measured by enzyme-linked im-munosorbent assay (ELISA). The expression levels of IL-23, RORγt and IL-17 mRNA in skin allografts were detected by re-al-time fluorescent quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results Compared with con-trol group, the average survival time of mouse skin allografts was significantly prolonged in low-dose group and high-dose group (P＜0.05). The less lymphocyte infiltration and destruction of architecture were found in the skin biopsies. The serum expression of IL-23 protein was lower (P＜0.05), but no significant difference was found in two treatment groups. The serum expression levels of IL-17 protein were reduced in turn in receptors of control group, low-dose group and high-dose group (P ＜ 0.05). The expression levels of IL-23, RORγt and IL-17 mRNA in skin grafts were significantly lower in low-dose group and high-dose group than those of control group (P＜0.05), but no significant difference was found in two treatment groups. Conclusion ATRA can effectively prolong the survival time of skin allografts, which may related with the inhibi-tion of the expression of IL-23, RORγt and IL-17 mRNA and the development of IL-23 and IL-17 protein.

Objective:To optimize the extraction technology of compound Xiongdan capsules. Methods: The effects of water a-mount, decoction duration and decoction times on the water extraction technology were investigated by orthogonal design using the ex-tract derivation rate and the content of ginsenoside Rg1 , Re and Rb1 as the indices. Results:The optimal water extraction conditions for ginseng and Nidus vespa were as follows:decocting three times with 8-fold of water, and extracting for one hour each time. The extract was condensed until the relative density was 1.20(80-90℃), and then dried and crushed into fine powder. The other herbs were crushed into fine powder and screened by No. 5 meshes. The above powder was blended and well mixed with starch, and then packed into capsules. Conclusion:The optimized water extraction technology is stable, feasible and reproducible, which provide reference for industrial production.

Objective To screen the optimum conditions carrying murine melanoma B16 cells in spaceflights without cares.MethodsMurine melanoma cells were cultured on micocarriers and grouped depending on cells concentration, serum concentration, microcarrier number and temperature.After 33 days, B16 cells were stained by Giemsa, observed with phase-contrast microscope and counted for surviving percentage.ResultsThe optimum conditions,in which the surviving percentages were 8% and 10%, were obtained in the experiments.B16 cells were carried in the 20th recoverable satellite orbiting 18 days under the optimum conditions. After recovering, 110 strain monocloned cells were survived and the surviving percentage was 1.1%.ConclusionThe optimum conditions carrying murine melanoma B16 cells in spaceflights without cares seems to be obtained,and it did improve the surviving time and percentage of cells in spaceflights without cares.