Objective To establish a novel flow cytometric crossmatch assay allowing detection of complement-fixing donor specific anti-HLA IgG alloantibodies (Flow cytometry complement-dependent crossmatch,Flow-CDC). Methods One hundred pretransplant crossmatchings were performed using Flow-CDC and NIH-CDC between 62 patients awaiting renal transplantation and 33 donor cells.These crossmatchings were divided into two groups according to PRA.Group 1 consisted of 25 sera with negative PRA,and group 2 consisted of 75 sera with positive PRA.All of the sera were pretreated with DTT to inactivate IgM. Lymphocytes were isolated from peripheral blood (or,in a few instances,from the spleen) of the cadaveric donors. The correlation between different techniques for detection of donor specific anti-HLA antibodies was evaluated.The effect of both methods on clinical transplantation outcome was observed. Results In group 1,NIH-CDC and Flow-CDC were negative for all 25 sera.In group 2,24 (32.0%) had a positive NIH-CDC,31 (41.3%) had a positive Flow-CDC.There was a significant difference between two methods (?2=5.14, P= 0.016 ).Overall concordance between both tests was 93% with 69 concordant negatives and 24 concordant positives. The correlation coefficient (r) was 0.80.In group 2,5 patients received transplantation. One of them with negative NIH-CDC and positive Flow-CDC suffered from acute rejection after transplantation and lost the graft,and the other patients with negative NIH-CDC and Flow-CDC had good outcome. Conclusions Flow-CDC can detect specifically complement-fixing IgG alloantibodies against donor HLA and is more sensitive than NIH-CDC.Additionally,the computer printouts represent a permanent record of the crossmatch for retrospective review.Flow-CDC may become the standard crossmatch method as a possible alternative to conventional NIH-CDC testing.

Objective To study the antitumor activity of steroid saponins,such as Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin,extracted from Paris polyphylla var.yunnanensis against lung adenocarcinoma cells.Methods Taking mouse lung adenocarcinoma LA795 cell line and its T739-bearing mice as models,the antitumor activity of steroid saponins of P.polyphylla var.yunnanensis against lung adenocarcinoma cells was carried out in vitro and in vivo.The inhibitory activity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin on the LA795 cell line was studied by means of MTT assay and their antitumor effects in vivo were carried on inbred strains of mice(T739) affected by LA795 metastatic lung cancer.Results The MTT assay of in vitro cytotoxicity of Rhizoma Paridis saponins,Rhizoma Paridis saponin Ⅰ,and diosgenin showed strong cytotoxicity against the growth of LA795 cells.IC50 Values were 24.33 ?g/mL,1.85,and 149.75 ?mol/L,respectively.The tumor weight in all treated groups was significantly lower than that in the control group and the inhibition of tumor growth were 41.82%,29.44%,and 33.94% in their high-dose groups.The metastasis of tumor cells was inhibited by different degrees in treated groups.Conclusion Steroid saponins of P.polyphylla var.yunnanensis have tumoricidal activity on lung adenocarcinoma cell line,both in vitro and in vivo.

Objective To identify the changes of outpatients/emergency visits at Nanjing Children′s Hospital since introduction of the general pricing reform at public hospitals.Methods Fluctuations of outpatients/emergency visits at Nanjing Children′s Hospital before and after the reform in 2015 were compared,and an empirical analysis was made in view of the overall structural changes,demand price elasticity and cross-price elasticity of various outpatients/emergency visits.Results The change of total outpatients/emergency visits was minimal,yet the composition registered significant changes.Monthly average of common outpatients totaled 7 072 visits,a growth of 12.6%;that of senior specialists totaled 3 035 visits,a growth of 48.4%.The price elasticity was<1,while the demand for various outpatients/emergency visits services and cross-price elasticity appeared inelastic.Conclusions The one year reform has witnessed the hospital in smooth operations as yet.Given the sizable rise of examination prices for both outpatients/emergency visits,patient volume remains stable instead of drastic decrease,evidencing the strong demand for children′s medical services.

Anti-glutamic acid decarboxylase antibody-associated encephalitis is a kind of autoimmune encephalitis mediated by anti-glutamic acid decarboxylase antibody, which belongs to anti-neuronal intracellular synaptic protein antibody-associated encephalitis. Clinical manifestations include stiff-person syndrome, cerebellar ataxia, limbic encephalitis, seizures, etc., often associated with a variety of autoimmune diseases, rarely associated with tumors. Detection of anti-glutamic acid decarboxylase antibody is crucial for clinical diagnosis. Immunotherapy helps to relieve symptoms and improve prognosis. The incidence of this disease is low, and there are few reports at home and abroad. This paper intends to review the research on this encephalitis, hoping to improve the clinicians′ understanding and the level of diagnosis and treatment of the disease.

Background Small incision lenticule extraction (SMILE) was a new procedure which is miniinvasion and flapless,but few attentions were paid to the visual quality after SMILE.To understand the changes of intraocular straylight after SMILE is of important significance for the assession of the visual quality after the procedure.Objective This study was to compare the characteristic of straylight between SMILE and femtosecond laser assisted laser in situ keratomileusis (FS-LAISK) after surgery.Methods A retrospective study was designed.Written informed consent was obtained from each patient prior to accepting the surgery.One hundred and twenty-seven eyes of 68 patients with myopia and myopic astigmatism were included in Tianjin Eye Hospital from January 2011 to July 2013.SMILE was performed on 64 eyes of 35 patients and FS-LASIK was carried out in 63 eyes of 33 patients at the same period.Stray light was detected with C-Quant stray light meter before and 1 week,1 month and 6 months after surgery.The changes of stray light values following the surgery were compared between the two groups,and the correlations between postoperative stray light values with operative parameters were analyzed.Results The mean stray light values were (1.03 ±0.19)D and (0.95±0.16)D at 1 week and 1 month after surgery,which were higher than (0.88 ±0.18)D at preoperation in the FS-LASIK group (P =0.000,0.012).The stray light values were (0.98±0.16),(0.95±0.14) and (0.94±0.16)D at 1 week,1 month or 6 months after surgery,and no statistically significant differences were found in comparison with (0.91 ±0.15)D in preoperation in the SMILE group (all at P＞0.05).No significant intergroup difference was seen in the stray light values (Fgroup =0.077,P=0.781).The stray light differences of various time points after surgery and before surgery were increased in the FS-LASIK group compared with SMILE group (Fgroup =14.798,P＜0.001),with the significant difference in postoperative 1 week between the two groups (P＜0.01).The negative correlations were found between stray light values at 1 week,1 month and 6 months after SMILE with the lenticule thickness/central corneal thickness (CCT) (R2=0.123,0.145,0.098,all at P＜0.05) or between stray light change.Conclusions Intraocular stray light appears less change in operative eyes after SMILE.However,intraocular stray light increases in the eyes received FS-LASIK,especially at early stage after operation.The stray light shifts return to normal with the lapse of postoperative time.

Objective To investigate the statistical methods used in nursing papers for bachelor's degree. Method We collected 160 nursing theses for bachelor's degree in 2016 in our college and analyze the statistical methods. Results A total of 80 papers used statistical methods consisting of statistical table, statistical inference, single factor analysis, variance analysis and statistical description. Among them, the statistical methods in 66 papers (82.5%) were misused. About 139 misuses of statistical tables was found in papers, and about 23 misuses of statistical inference in papers, taking up 45.4%. Conclusions The nursing undergraduates are familiar with the application of statistical methods, but the methods tend to be simple and unitary. Therefore, it is a need to enhance their practice in using statistical methods.

Objective To investigate the protective role of hydrogen sulfide and the expression of cystathionine gamma-lyase/hydrogen sulfide pathway in a mouse model of myocarditis induced by Coxsachie -virus B3(CVB3).Methods A total of 110 five-week-old BALB/c male mice were randomly divided into four groups:the control group, viral myocarditis group, sodium bisulfide (NaHS) group (50 μmol/kg) and DL-propargylglycine (PAG) group (40 mg/kg).The experimental model of viral myocarditis was induced by intraperitoneal injection of CVB 3.Then the four groups were respectively administered with PBS , PBS, NaHS and PAG from day 1 to day 10 after infection.Blood and heart specimens were harvested from 10 mice of each group on day 4 and day 10 for evaluation of myocardial edema .The pathological changes in heart tis-sues were observed through hematoxylin-eosin staining.Levels of H2 S, IL-6 and TNF-αwere measured by ELISA.The expressions of CSE and CVB 3 at mRNA level were determined by quantitative real time PCR ( qRT -PCR ) analysis and the expression of CSE at protein level was detected by Western blot .Results Compared with the control group , the levels of H2 S and the expressions of CSE at mRNA and protein levels were down-regulated in mice with CVB 3-induced myocarditis .With the treatment of NaHS , the levels of H 2 S in serum and tissue were both up-regulated , and the histopathological damage was alleviated .However , PAG as an irreversible CSE inhibitor inhibited the expressions of H 2 S and CSE and aggravated myocardial injury , inflammatory cells infiltration and interstitial edema .Moreover , the RT-PCR analysis also showed that the expression of CVB3 at mRNA level was inhibited by NaHS but enhanced by PAG .Conclusion The expres-sion of CSE/H2 S pathway is down-regulated in mice with CVB 3-induced viral myocarditis .PAG could pro-mote virus propagation and exacerbate the disease through inhibiting the production of endogenous H 2 S, while NaHS as a H2 S donor has a protective effect on infected myocardium by suppressing virus replication at an early stage .

Objective To establish and evaluate the new method of 8-color flow cytometry (8c-FCM) with two tube detecting bone marrow minimal residual disease (MRD) in B lineage acute lymphoblastic leukemia (B-ALL).MethodsThe MRD cells were analyzed by using two combinations of 8c-FCM antibody panels,gating with CD19/Side scatter(SSC),CD45/CD10 and CD34(or cTdT).Nalm-6 cell of B-ALL was mixed into normal marrow cells,with proportion of 10.00％,1.00％,0.10％,0.01％and 0.005％,and recovery test and reproducibility test were carried with 8c-FCM established to value its accuracy and precision of.Fluorescence intensity was detected on different time points after marked Nalm-6 by antibodies to evaluate the fluorescent stability of the antibodies.The immunophenotyping was analyzed in 39 bone marrow specimens,including 9 cases of normal control,9 cases of B-ALL primary or recurrent,21 cases of complete remission (CR) after chemotherapy or bone marrow transplantation ( BMT),to evaluate the detection of normal B lymphocyte lineage,leukemia cells of B-ALL,MRD cells of B-ALL using the 8c-FCM and compare it with 4c-FCM in being.ResultsThe method of two tube 8c-FCM with main antibodies of CD19,CD45 and CD10 to detect MRD of B-ALL was founded; CV ＜ 2.5％,average recovery rate was 95.81％,when the actual percent of Nalm-6 mixed into normal bone marrow≥0.10％,and the percent of Nalm-6 detected and actual was linear dependent ( r =0.99,P ＜ 0.05 ) ranged from 10.00％ - 0.01 ％,when 106 cells were acquired ; the sensitivity of the method established could reach 0.01％.The fluorescent intensity decreased along with the time after Nalm-6 cell marked,but less than 10％ in 24 hours.Using the antibody combinations and analyze strategy,the immunophenotye of B lymphocyte in normal bone marrow presented four sequential stages:Stage Ⅰ CD45low/CD10stro/CD20 -/CD38stro/CD34 + or cTdT +,Stage ⅡCD45 +/CD10 + / CD20 -/CD38stro/CD34+ or cTdT +,Stage Ⅲ CD45 +/CD10 +/CD20 -/CD38stro/CD34 -or cTdT-,Stage Ⅳ CD45stro/CD10 -/CD20 +/CD38low/CD34or cTdT.Antigen expressions of leukeamic cells of B-ALL primary or recurrent were different compared with control team; there were 5 cases with MRD positive in CR team,and the main antigen expression was consistent with the results from 4c-FCM.The range of the percent of MRD cells was 0.02％ - 5.42％ of the 5 cases of MRD positive.ConclusionsThe new method of two tube 8c-FCM established shows good reproducibility and high accuracy,and can identify normal B lymphocyte populations in bone marrow and regenerated B-precursors in CR cases with MRD cells;compared with 4c-FCM,the new method of two tube 8c-FCM with the fewer specimen is faster and efficient to diagnose MRD of B-ALL.

[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.

Objective: To study the effect of cytochrome P-450 4F2 (CYP4F2, rs2108622) gene polymorphisms in patients with warfarin for initial doses in 7 days.
Methods: A total of 271 patients treated by warfarin were studied. The CYP4F2 gene polymorphisms were assessed by real-time PCR, the average initial warfarin doses in 7 days and the time of international normalized ratio (INR) ifrst arrived to therapeutic range were recorded. The differences of initial warfarin doses and the time of INR ifrst arrived to therapeutic range among CYP4F2 gene polymorphisms of CC, CT and TT genotypes were analyzed by statistical method.
Results: The average initial warfarin doses among CYP4F2 polymorphisms of TT and CT/TT were higher than CC, P<0.05. The times of INR first arrived to therapeutic range in TT genotype was higher than CC genotype, P<0.05. With adjusted gender, age, body height and weight, CYP2C9 and VKORC1 polymorphisms, conditions of disease and medication, the effect of CYP4F2 gene polymorphisms on initial warfarin doses in 7 days were still with the statistic meaning (regression coefifcient=0.21, P=0.003).
Conclusion: CYP4F2 polymorphisms inlfuenced the initial warfarin doses in 7 days in relevant patients.