Objective To enhance the quality and efficiency of the outpatient cashier service.Methods A comprehensive outpatient cashier management system based on RFID was developed and put into use.Results The system could be used in outpatient cashier department for attendance checking,schedule management,bonus distribution,personnel management,data statistics and report printing.Conclusion The comprehensive outpatient cashier management system can improve the efficiency of the outpatient department.

Objective To explore the effects of single rectangle cage interbody fusion combined with pedicle screw fixa?tion for isthmic spondylolisthesis. Methods Data of the 34 cases with isthmic spondylolisthesis hospitalized at our department from March 2012 to February 2015 were retrospectively analyzed. All cases were operated by posterior pedicle screw fixation com?bined with single rectangle cage interbody fusion. There were 21 males and 13 females, aging from 18 to 63 years old (with an aver?age age of 41.6±8.2 years). Lesion segments were L4 in 15 cases and L5 in 19 cases;there were 12 cases in Meyerding I degree, 16 in II degree and 6 in III degree;28 patients had lower back pain associated with lower limb pain;19 patients had lower back pain associated with numbness of lower limb;18 patients had lower back pain associated with lower limb muscle weakness and 2 pa?tients had urination and defecation function disturbance. The surgical operation time, bleeding, postoperative drainage flow, graft fusion rate, complications were measured, and the intervertebral disc height, Japanese Orthopaedic Association (JOA), Oswestry disability indes (ODI), visual analogue scale (VAS) score were compared before and after treatment. Results The operation of 34 patients was successfully completed with the average operation time of 1.8 ± 0.8 h, and the bleeding in operation was 362.4 ± 93.7 ml, while postoperative drainage flow was 116.3±54.2 ml. All patients were effectively followed up for 12 months to 46 months (av?erage, one year and 10 months). Two weeks after operation, the intervertebral disc height were obviously increased (9.85±1.11 mm, 9.63 ± 1.04 mm), and there were significant difference compared with preoperative results (6.53 ± 0.98 mm, 6.24 ± 1.07 mm), but there were no significant difference compared with the latest follow?up results (9.23±1.33 mm, 9.30±1.16 mm). At the latest follow?up, JOA score was 13.19±1.08, ODI score 12.34±7.52, VAS score 2.23±1.51, and there were significantly different in each score compared with preoperative results. The symptoms of low back pain, lower limb nerve dysfunction and quality of life were signifi?cantly improved compared with preoperative situation. At the 16 month follow?up, all postoperative patients with vertebral body bone graft were judged to be fused. There was no pedicle screw loosening, fracture and so on. Conclusion Single rectangle cage interbody fusion combined with pedicle screw fixation for the treatment of isthmic spondylolisthesis is a kind of mini?invasive, less bleeding, safe and effective surgical method.

This article reviews and summarizes the research data about vertebrobasilar dolichoectasia in recent years from the aspects of epidemiology,pathogeny,pathology,diagnosis,clinical manifestations,as well as its mechanism and treatment.

BACKGROUND: At present, bone defects usually repaired by autologous bone, allogenic bone, synthetic bone substitutes and other methods, which received poor clinical results. Preliminary studies have shown that adipose-derived mesenchymal stem cells (ADSCs) possess strong proliferation ability and differentiation potential, and can be induced differentiate into bone. OBJECTIVE: To analyze the application of ADSCs in bone tissue engineering, and to identify whether ADSCs can be used as seed cells in bone tissue engineering. METHODS: The databases of PubMed (1999-01/2008-12) and Tongfang (2003-01/2008-12) was retrieved using key words of "adipose tissue-derived mesenchymal stem cells, adipose mesenchymal stem cells, adipose stem cell; osteogenic induction, osteogenic inducement, bone induction, osteoblastic induced; chondroblast induction, cartilage induction; bone tissue engineering, tissue engineering bone, tissue engineering of bone". RESULTS AND CONCLUSION: A total of 361 literatures were collected, including 246 in Chinese and 115 in English. Totally 29 literatures were accordant with the study criteria. ADSCs is a truly multi-directional differentiation potential cells, which possess strong amplification and self-renewal potential, and can be directional differentiated into osteoblasts, cartilage cells, bone cells and muscle cells. It can be used as seed cells in bone tissue engineering when matching appropriate stents.

Objective To clone and express recombinant outer membrane protein P6, determine its optimal expression conditions and to investigate the immunoprotective effects of the P6 protein on mice. Methods The P6 gene of nontypeable Haemophilus influertzae(NTHi) was amplified by PCR from the NTHi genome and cloned into expression vector pET-32a (+) to generate the pET-32a-P6 recombinants. They were confirmed by nuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). Its optimal expression conditions were determined such as engineering strains, the concentration of IPTG, inducing temperature, inducing time, different medium etc. The recombinant protein was purified by Q Sepharose~(TM) XL ion exchange and gel filtration chromatography. The protein was analyzed by SDS-PAGE, Western blot and sequencing. BALB/c mice were immunized with recombinant protein be-fore challenged by NTHi through intraperitoneal injection. Then the mortality rate of different group was com-pared. Results The recombinant P6 of NTHi was successfully constructed and expressed in E. coli at a rel-atively high level. The purity was up to 95% after purification. The relative molecular mass of the protein is 14 145. 848. The recombinant protein was confirmed to show specific reaction on the antiserum through Western blot. The animal experiments showed the mortality rates of immunization groups were significantly lower than that of the control group (P < 0.05). Conclusion The successful expression of the recombinant P6 will be very helpful for the further study on development of vaccine, its purified, immunological activity and antibody preparation.

ObjectiveTo detect the difference of cytokines and antibodies productions by immunologic system from mice and rabbits vaccinated with the M.RCAg-1 chimeric protein,expressed in E.coli,formulation with different adjuvants,including Freund's adjuvant and three clinically acceptable adjuvants,namely,Al(OH)3,Montanide ISA720 and Montanide ISA51.MethodsSix weeks female BALB/c mice were vaccinated with recombinant protein formulated with different adjuvants through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual Epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; Affinity assay between protein and immune IgG of rabbits with the biosensor,and the growth of Plasmodiumfalciparum in vitro to evaluate by growth inhibition assay(GIA).ResultsDifferent formulation can induce different levels of antibody titers,the effection of ISA51 adjuvant was most closely with Freund's adjuvant,and can induce a higher specific antibody of 11 epitopes within proteins,can effectively stimulate cellular immune response based on the IFN-γ,to avidity Montanide ISA51 adjuvant immune antibodies and M.RCAg-1 protein affinity than the other two adjuvants;and Montanide ISA720 adjuvants and Al (OH)3 adjuvant group in mice can't induce a significant IFN-γresponse(P＞0.05).On avidity assay,the Montanide ISA51 formulation group was better than the other two adjuvants; and Montanide ISA720 and Al (OH)3 adjuvant formulation group can't induce a significant IFN-γresponse in mice(P＞0.05) ; the inhibition rates were 60% and 100% in 3D7 and Dd2 Plasmodium falciparum at a concentration of 2 mg/ml IgG by Montanide ISA51 formulated protein,and IgG of Al( OH)3 formulation could not effectively inhibit the in vitro growth of Plasmodium falciparum( 10% ),while IgG of Montanide ISA720 formulation could not inhibit growth of parasite in vitro.ConclusionBy comparing three clinically acceptable adjuvants and Freund's adjuvant in BALB/c mice and New Zealand rabbit,Montanide ISA51 adjuvants can be acceptable formulated M.RCAg-1 protein induced humoral and cellular immune responses,can be used as one of the candidate adjuvants.

ObjectiveTo explore the effects of vector fusion peptides on the conformation and immune reactivity of recombinant polyepitope antigens,M.RCAg-1.MethodsWe subcloned polyepitope artificial antigen gene,M.RCAg-1,into prokaryotic expression vectors,pDS-ex,that contain different fusion tags at the N-terminus or no any tag by different restriction enzyme cutting site.Three recombinant proteins expressed by these vectors,named P312-1,P312-2,and P312-3,were purified and purity is greater than 95％.Then BALB/c mice were vaccinated with the three proteins formulated with Freund's adjuvant through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; and the growth of Plasmodium falciparum in vitro to evaluate by growth inhibition assay(GIA),and analyze secondary and tertiary structures of recombinant proteins from different expression vectors by bioinformatics and circular dichroism technique.ResultsThe P312-1,P312-2 have almost the same amino acid sequence,and the three proteins have the same immunogenicity in animal models(P＞0.05),however,the different proteins elicited various T-cell responses,the rabbit antibody induced by these proteins showed diverse efficacy in malaria parasite growth inhibition assaysin vitro ( respectively,93.9％,14.7％,54.3％ ).The significant differences of secondary and tertiary structures were shown in recombinant proteins from different expression vectors,analyzed by bioinformatics and circular dichroism technique,which demonstrated the change of protein molecule spaces conformation,and the obviously change of some epitope locations.ConclusionThese results suggest that the expressed polyepitope artificial antigens originating from the different vector fusion peptides indeed affect the protein folding and,subsequently,the epitope exposure.Thus,these proteins are able to induce both distinct humoral and cellular immune responses in animal models,and they affect the efficacy of immune inhibition against the parasite,the enhancement or suppresses.

Objective To construct a stable infectious clone of S191 virus strain used for the pro-duction of live attenuated measles vaccine. Methods Full length cDNA of S191 strain and gene fragments encoding nucleocapsid(N),phosphoprotein(P)and RNA polymerase(L)were synthesis and respectively cloned into the vector pVAX1. The 293T cells were respectively transfected with the recombinant expression plasmids and co-cultured with Vero cells. The supernatants of cell culture were collected for identifying res-cued viruses. The indirect immunofluorescence assay was performed for virus identification. The rescued viruses at different passages in Vero cells and the sequences of the rescued viruses were analyzed. Results Restriction enzyme digestion and sequence analysis showed that the recombinant expression plasmids contai-ning the full length cDNA with an artificially engineered mutation at nucleotide 2101(C-A)and gene frag-ments encoding N,P and L proteins of S191 strain were constructed successfully. The N and P proteins were detected in Vero cells with immunofluorescence assay. A cytopathogenic effect on Vero cells was induced by rescued viruses. Conclusion The stable infectious clones of S191 virus used for the production of live at-tenuated measles vaccine were rescued successfully. An approach by using reverse genetics technique for S191 strain study was established which could be used for the development of new chimeric vaccines against measles virus.

Objective To explore the clinical features, diagnosis, and treatment of congenital disorder of glycosylation type 1a (CDG-Ⅰa), a rare inherited metabolic disease. Methods The clinical data and the gene detection results of one case of CDG-Ia which was discovered because the case had encephalopathy and hepatopathy were retrospectively analyzed. The related literatures were reviewed. Results Male infant suffered with face and trunk rash, motor development retardation, malnutrition, cheek fat plump, low limbs muscle tone, and bilateral crater nipple at 3 months old. Abnormal liver function and mild renal impairment were found after examination. The development quotient was low. Head MRI showed that bilateral frontal and temporal sulcus widening, and cerebellar atrophy. Urinary organic acids, amino acids, carnitine, and biotin activities were normal. Gene sequencing revealed that there were two heterozygous mutations, c.430T>C (p.F144L) and c.713G>C (p.R238P), in the PMM2 gene. The diagnosis of CDG-Ⅰa was confirmed. Both of the infant's parents were healthy, and each of them carries a pathogenic mutation. The infant had an elder brother who had mental disorder and died for liver and kidney function damage and hydronephrosis at 8 months old. Conclusion CDG-Ⅰa is an autosomal recessive disease. For infants with unexplained multiple organ damage, especially combined with intelligent and motor development retardations, strabismus, nipple retraction, and cerebellar atrophy, the possibility of CDG-Ⅰa should be considered. Gene detection of PMM2 can help the diagnosis.

Recent research data on post-stroke seizures and epilepsy are reviewed and summarized.They are reviewed from the aspects of concept, risk factors, epidemiology, pathogenesis, clinical manifestations, diagnosis, as well as prevention and treatment.