With the advancement of medical and health system,the reform of medical price became one of cores in new medical reform.According to the principle of "the total quantity control,the structure adjustment",in recent years,every region adjusted the medical price.However,the regional medical price disharmony was influenced by the mechanism of information-sharing,the differentiation right of pricing and management and setting basis and cycle.It followed the regional medical price disharmony concern over the principle,content and method of mechanism of regional medical price coordination for optimizing medical price management,so as to provide references for further optimizing medical service price management.

Objective To explo the antioxidant effect and molecular mechanism of gastrodin (Gas) in epilepsy (EP) rats induced by LiCl-pilocarpine (PILO) . Methods Eighty male SD rats were randomly divided into 5 groups: sham group, EP group, therapy groups (pretreated with 60 mg/kg, 90 mg/kg, 120 mg/kg of gastrodin respectively) . The EP model was esteblished by peritoneal injection of LiCl-PILO. Therapy groups were pretreated with various concerntration of Gas. The control group was given the same dosage of normal saline. The alteration of behavior was observed, the concentration of catalase (CAT), glutathion (GSH), superoxide dismutase (SOD), glutathion reductase (GR), total antioxidtion (T-AOC) and malondialdehyde (MDA) in rats brain cortex were detected by chemical colorimetric method, phosphorylation of p38 was determined by western blot. Results There was no EP seizure in sham group,and the EP seizure degree in therapy groups (gas pretreated groups) was significantly decreased,and had statistically significant difference with EP group (P<0.05) . The EP model rats exhibited a significant decrease in the concentration of endogenous antioxidants (CAT, GSH, SOD, GR and T-AOC), while an increase of the concentration of MDA and phosphorylation p38 protein as compared to sham group (P<0.05) . After treatment of the Gas,treatment group rats attenuated the seizure degree,exhibited a significant increase of the concentration of endogenous antioxidants (P<0.05),while a decrease in concentration of MDA and phosphorylation of p38 as compared to model group (P<0.05) . Conclusion Gas may have a neuroprotective role in central nervous system of epileptic rats modle by down-regulateing the seizure degree and the activity of p38 kinase and up-regulateing the content of endogenous antioxidants.

Objective To explore the activition and polarization of microglia in the epileptic rats induced by lithium chloride-pilocarpine. Methods One hundred male SD rats were randomly divided into five groups: control group and different time points model groups including 1d,3d,7d and 14d. Epilepsy models were established by lithium chloride-pilocarpine intraperitoneal injection. The control group was given the same dosage of normal saline. The morphology change was detected by immunofluorescence,and the expressions of iNOS and Arg-1 were determined by IHC at respective time points. Results Compared the model groups with control group,microglia was activated,synapsis was shorten,volume got bigger,most of them seemed as amoebocyte,the expression of iNOS increased and Arg-1 decreased,especially at 3d.ConclusionThe results from this study indicated that microglia was activated and polarized in epileptic rats induced by pilocarpine.

The expression of Fas ligand (FasL) on the membrane of many kinds of leukemia or solid tumor cells played an important role in the immune escape of tumor cells. This study was aimed to know if the soluble Fas (sFas), expressed by adenovirus, could block the immune escape of tumor cells by FasL pathway. The two recombinant adenoviral vectors, AdsFas with murine soluble Fas gene and AdEGFP with enhanced GFP protein gene, were constructed by homologous recombination between two plasmids in Escherichia coli with the AdEasy adenovirus vector system. The viruses were propagated and purified by two times ultracentrifugation. Their titres were detected by plaque assays. The expressed protein was evaluated by Western blot analysis. Then the tumor EL4 cells were infected with AdsFas and AdEGFP respectively. The apoptosis ratio of the target cells-YAC-1 cells induced by EL4 cells was respectively detected by (3)H-thymidine ((3)H-TdR) labeling. The results showed that the recombinant adenoviral vectors AdsFas and AdEGFP were successfully obtained. The titres of viruses purified by two times ultracentrifugation were up to 10(11) pfu/ml by plaque assays. The sFas protein was highly expressed in the target cells by Western blot analysis. After the EL4 cells were transfected with the adenoviruses AdsFas, the apoptosis rate of YAC-1 cells in the sFas transfection group (respectively 6%, 7% and 9% when the effector:target (E:T) was 3:1, 10:1 and 30:1) was obviously lower than that in the control group (respectively 28%, 37% and 45%), P < 0.01. But when the EL4 cells were transfected with AdEGFP, the apoptosis rate of YAC-1 cells (respectively 30%, 36% and 48%) was similar to the control group, P > 0.05. In conclusion, the transfer of sFas by adenovirus could inhibit the apoptosis of Fas(+) cells-YAC-1 cells induced by tumor EL4 cells. It showed that the transduction of sFas could block the effect of the immune escape of EL4 cells through FasL in vitro. These results thus provide a new direction to find a way to treat tumors.
Adenoviridae ; genetics ; Animals ; Apoptosis ; Blotting, Western ; Fas Ligand Protein ; Leukemia, T-Cell ; immunology ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; Mice, Inbred C57BL ; Transfection

OBJECTIVETo investigate whether murine soluble Fas gene transfected marrow graft could block the immune escape of leukemia cells, so as to eliminate the residual leukemia cells and reduce relapse after bone marrow transplantation (BMT).

METHODSThe murine leukemia/lymphoma models were established by inoculating female C57BL/6 mice (H-2b) with 10(5) EL4 cells/mouse through caudal vein. Donors of BM grafts were C57BL/6 male mice. Bone marrow mononuclear cells (BMMCs) were transfected with sFas or EGFP by adenovirus (adsFas or adEGFP) 24 hours before BMT (group D or E). The following three groups were set simultaneously: group A, no BMMCs transplanted; group B, BMMCs transplanted with no adenoviruses transfection; group C, EL4 cells transfusion only. Hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all the groups after BMT.

RESULTSThe spleen indices examined 11 days after BMT were not obviously different among group B, D and E (P > 0.05), but in group A were significantly lower than those in the groups B, D, E (P < 0.01). The leukocyte and platelet counts on day 30 after BMT were recovered in group B and D, but were very low in group C and E. The Y-chromosomes appeared 2 months after BMT. Bone marrow pictures in group B and D were almost normal, but in group C and E had plenty of lymphoblast-like tumor cells. Tumors were obviously revealed in the mice of group C and E by histopathology examination, but did not in group B and D. The survival rate was 0 in group A, 100% in group B and D, 12.5% in group C and 6.25% in group E. Compared with that in group E, the survival was significantly increased in the sFas group (P < 0.01).

CONCLUSIONSGraft transfected with sFas gene prolonged the post-BMT survival of leukemia/lymphoma mice. The transfection of sFas might block the effect of the immune escape of EL4 cells through FasL.

Animals ; Bone Marrow Transplantation ; immunology ; Combined Modality Therapy ; Female ; Genetic Therapy ; methods ; Leukemia, Experimental ; immunology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Transduction, Genetic ; Transfection ; Transplantation, Homologous ; Tumor Escape ; fas Receptor ; genetics

Staphylococcus aureus is one of the most frequently encountered zoonotic pathogens.This bacterium produces the notable virulence factors such as hemolysin,panton-valentine leucocidin,exfoliative toxins and enterotoxin,which can cause invasive disease in humans and animals.Methicillin-resistant S.aureus (MRSA) is a multidrug-resistant bacterium which acquired the staphylococcal chromosome cassette mec (SCCmec).SCCmec is one of the key reasons for the antibiotic resistance of MRSA.As for MRSA resistance,the β-1actam resistance is mediated by mecA gene,and the drug-resistance genes inserted in the variable area of the SCCmec element play an important role in the multidrug resistance of MRSA.In recent years,it has been reported in Europe,North America and other countries that the multidrug resistance MRSA was detected in aquaculture environment and livestock.Besides,MRSA poses a serious threat to public health,and it can colonize and cause invasive disease in humans through aquaculture environment or other ways.This review summarizes drug resistance change of S.aureus and analysis of SCCmec resistance elements,toxicity and prevalence of livestock-associate MRSA,which would have theoretical and practical significance to understand S.aureus drug resistance,SCCmec typing,as well as control and prevent LA-MRSA transmission and infection between animals and humans.

Leukemic cells from patients expressed high level FasL cause apoptosis of autologous activated T cells via the Fas/FasL pathway. To investigate the role of soluble Fas (sFas) in reversing this process, a retroviral-mediated expression vector pLXIN-sFas was established. A retroviral-mediated expression system of human sFas was established in vitro and the biological activity of the expression product sFas was observed. To obtain the soluble Fas cDNA, the specific part of the full-length Fas cDNA was deleted by multiple PCR. After pLXIN-sFas packaged by PA317 cells, it was transferred into the target cell COS-7. The quantity of the sFas was (2.2 +/- 0.7) micro g/ml in supernatant of cultured COS-7 cells, and it could greatly inhibit apoptosis of Jurket cells induced by anti-Fas antibody. Our results suggested that the recombinant is able to express the target proteins in vitro and it has the perfect biological activity.
3T3 Cells ; Animals ; Apoptosis ; drug effects ; COS Cells ; Cell Line ; Cell Survival ; drug effects ; Culture Media, Conditioned ; pharmacology ; Dose-Response Relationship, Drug ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Jurkat Cells ; Mice ; Retroviridae ; genetics ; Solubility ; fas Receptor ; genetics ; metabolism ; pharmacology

OBJECTIVETo evaluate the effect on pre-exposure prophylaxis (PrEP) to prevent HIV infection in high risk populations.

METHODSA computerized literature searching had been carried out in PubMed, EMbase, Ovid, Web of Science, Science Direct, Wanfang, Tsinghua Tongfang database and related websites to collect relevant papers (from establishment to June 2012) with the key words of pre-exposure prophylaxis, HIV, AIDS, high risk populations, relative risk, reduction. All randomized controlled trials (RCT) papers about using single or compound antiretroviral drugs (ARVs) orally or topically before HIV exposure or during HIV exposure in high risk populations were enrolled. Meta-analysis was conducted using Stata 10.0 to calculate the pooled RR value (95%CI). Consistency test was performed and publication bias was evaluated.

RESULTSFinally 5 RCT papers were enrolled, including 10 271 persons who were at high risk of HIV infection. The number of the experimental group was 5929, among which 116(1.96%) became infected. The number of the control group was 4342, among which 201(4.63%) became infected. Meta-analysis showed that the pooled relative risk (RR) and 95%CI was 0.49 (0.39 - 0.61), P < 0.05, indicating that the persons in experimental group had a 0.49 times lower risk of HIV infected, as compared with the control group. Publication bias analysis revealed a symmetry funnel plot. The fail-safe number was 825.

CONCLUSIONPrEP was an effective and safe protection measure to reduce HIV infection in high risk populations.

Anti-HIV Agents ; administration & dosage ; therapeutic use ; HIV Infections ; prevention & control ; Humans ; Randomized Controlled Trials as Topic ; Risk

BACKGROUNDTo investigate if bone marrow transplantation (BMT) with bone marrow mononuclear cells (BMMCs) transducted with murine soluble Fas gene (sFas) using adenovirus vector could block the immune escape of leukemia cells eliminate the residual leukemia cells and reduce their relapse.

METHODSThe recombinant adenovirus vector with murine sFas, adsFas, and the control vector adEGFP were constructed using homologous recombination between two plasmids in Escherichia coli. BMT was carried out after the BMMCs were infected with Adenoviruses. The mice models of leukemia/lymphoma were constructed by inoculating female C57BL/6 mice (H-2b) with 10(5) EL4 cells/mouse through caudal vein. Donors of bone marrow grafts were syngeneic male mice. BMMCs were infected with AdsFas or AdEGFP 24 hours before (Group D or E). The following three groups were simultaneously used: Group A, no BMMCs transplanted; Group B, transplanted with BMMCs not infected with adenoviruses; Group C, only transfusing EL4 cells, neither irradiation nor BMT. The hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all groups after BMT.

RESULTSThe adenovirus vectors were successfully constructed. The titre of virus after purification was up to 2.5 x 10(11) pfu/ml. Spleen indices examined 11 days after BMT were not obviously different among Group B, D and E (P > 0.05), but indices in Group A were significantly lower than those in the latter three groups (P < 0.01). Counts of leukocytes and platelets on +30 day showed mice were reconstituted satisfactorily in Group B and D, but very low in Group C and E. The Y-chromosomes existed 2 months after BMT and examination of bone marrow cytology showed that Group B and D were almost normal, but Group C and E had plenty of lymphoblast-like tumor cells. Tumors were obviously observed in the mice of Group C and E by histopathological examination, but the mice in Group B and D were normal. The survival rates were 0 (0/4) in Group A, 100% in Group B (6/6) and D (16/16), 12.5% (2/16) in Group C and 6.25% (1/16) in Group E respectively. It is demonstrated that, in contrast with the control (Group EGFP), survival rate was significantly increased in the sFas Group (P < 0.01).

CONCLUSIONSThe transfer of sFas gene by adenovirus changed the prognosis state of leukemia/lymphoma mice after auto-BMT. The transduction of sFas might block the effect of the immune escape of EL4 cells through FasL. These results could thus provide a new direction to find a way to treat the leukemia and its recurrence after BMT.

Adenoviridae ; Animals ; Bone Marrow Transplantation ; Fas Ligand Protein ; Female ; Genetic Vectors ; Leukemia, Experimental ; Leukocytes, Mononuclear ; Male ; Membrane Glycoproteins ; genetics ; Mice ; Mice, Inbred C57BL ; Recombination, Genetic ; Transduction, Genetic ; Transfection ; Tumor Escape ; physiology

To determine whether the Fas receptor-Fas ligand (FasR-FasL) system, which triggers apoptosis in sensitive cells, is an important mechanism of cytotoxicity in myeloid leukemia. FasL expression was investigated in myeloid leukemia cells and its upregulation by a combination of IL-2 and INF-gamma, +/- as well as its function of inducing Jurkat cells to apoptosis mainly by flow cytometry (FCM). Results showed that leukemia cells expressed more FasL (3.59 +/- 1.05)% than that expressed in the healthy individuals (0.36% +/- 0.16)%, P < 0.001 and the FasL was upregulated (7.78 +/- 3.40)%, P < 0.01 when treated with IL-2 and IFN-gamma. Leukemia cells were co-cultivated with Jurkat cells for 24 hours. Then Jurkat cells were labeled with FITC-annexin V and PE-CD3 to assess apoptosis by FCM. The leukemia cells, which had been incubated with IL-2 and IFN-gamma, induced more Jurkat cells to apoptosis than the ones that freshly isolated from the peripheral blood mononuclear cells, which raised the figure from (8.28 +/- 1.61)% to (10.73 +/- 2.16%). And the supernatant derived from the former killed more Jurkat cells than the latter. It was concluded that human myeloid leukemia cells expressed high levels of functional FasL that can kill Jurkat T-cells by apoptosis. FasR-FasL sys tem could play a role in the "immune escape" and relapse of the leukemia. The induction of apoptosis through the Fas pathway might be a novel and effective approach for leukemia immunotherapy.
Adolescent ; Adult ; Apoptosis ; genetics ; Child ; Child, Preschool ; DNA, Neoplasm ; genetics ; Fas Ligand Protein ; Female ; Flow Cytometry ; Humans ; Jurkat Cells ; Leukemia, Myeloid ; genetics ; metabolism ; pathology ; Male ; Membrane Glycoproteins ; biosynthesis ; Middle Aged ; Tumor Cells, Cultured