95%).Strains of PNSP were significantly more common than those PSSP(72.56% vs 22.72%),and strains of Sp resistant to erythromycin were significantly more common than susceptible to erythromycin too(94.42% vs 0.88%).Conclusions The antimicrobial resistance of Sp from children with lower respiratory tract infection in Wenzhou area has became a severe problem.Multi-drug resistance of Sp has been observed to chlorodeoxylincomycin,erythromycin,tetracycline and so on.The results will provide some important information for the therapy and the choice of antibiotics of Sp infection.

Objective To investigate the expression of Yes‐associated protein(YAP) in human papillary thyroid cancer and its influence on cell growth .Methods The samples in 57 cases of papillary thyroid cancer treated by operation resection in the gen‐eral surgery department of this hospital and the matched tumor‐adjacent tissues were collected .All the cases were definitely diag‐nosed by the pathology examination .The expression of YAP protein in the cancer tissue and corresponding tumor‐adjacent tissue were determined by the immunohistochemistry(IHC)staining .The relationship between the YAP protein and the clinicopathological data was statistically analyzed .siRNA was used to silence the expression of YAP in B‐CPAP cells ,MTT and flow cytometry were used to measure the proliferation and apoptosis changes .Results The expression level of YAP was markedly higher in papillary thyroid cancer tissues than in tumor adjacent tissues (P<0 .05);moreover the expression of YAP protein was positively correlated with the tumor size and TNM stage (P<0 .05) .Silencing YAP gene could significantly inhibit the cell proliferation ability and pro‐mote cell apoptosis(P<0 .05) .Conclusion YAP is highly expressed in papillary thyroid cancer tissues and is related with its ad‐verse clinicopathological characteristics ,down‐regulating YAP gene can significantly inhibit the cell growth .

Objective To investigate the relationship between the expression of CC chemokine ligand 5(CCL5)and S100 calcium binding protein A4 (S100A4)protein in breast cancer tissues with clinicopathological features and prognosis.Methods The expression of CCL5 and S100A4 in 40 cases of normal breast tissues and 120 cases of breast cancer were detected by immunohistochemistry,and the relationship between the degree of expres-sion and the clinicopathological features and prognosis of breast cancer was analyzed.Results The expression posi-tive rates of CCL5 and S100A4 in breast cancer tissues were 56.67% and 62.50% respectively,which were not expressed in normal breast tissues,and the differences were statistically significant (χ2CCL5 =39.403,P <0.01;χ2S100A4 =47.062,P <0.01).The expression of CCL5 in breast cancer tissues was statistically correlated with clinical staging(χ2 =10.141,P <0.01 ),pathological type (χ2 =5.769,P =0.017)and lymph node metastasis (χ2 =34.178,P <0.01),but not correlated with patients'age,tumor size,pathological grading(all P >0.05 ).And the expression of S100A4 was statistically correlated with clinical staging(χ2 =44.311,P <0.01)and lymph node metas-tasis(χ2 =14.843,P <0.01 ),but not correlated with patients'age,tumor size,pathological type and pathological grading(all P >0.05).The expression of CCL5 and S100A4 in breast cancer was positively correlated(r =0.301, P <0.01).CCL5 was positively correlated with the recurrence of breast cancer(OR =6.270,P <0.01),and S100A4 was not correlated with the recurrence of breast cancer(OR =1.103,P =0.768).Survival analysis showed that the disease -free survival time of patients with positive CCL5 expression was significantly shorter than the patients with negative CCL5 expression(χ2 =11.851,P <0.01 ),and the disease -free survival time of patients with positive S100A4 expression was significantly shorter than the patients with negative S100A4 expression(χ2 =5.433,P =0.021).The joint detection showed that the disease -free survival time in CCL5(+)+S100A4(+)group was sig-nificantly lower than that of CCL5(+)or S100A4(+)group(χ2 =15.341,P <0.01)and CCL5 (-)+S100A4 (-)group(χ2 =15.341,P <0.01).Conclusion The expression of CCL5 and S100A4 in breast cancer can reflect the metastasis and staging of breast cancer,which can be used to judge the clinical pathological characteristics and prognosis of breast cancer.

Objective To evaluate the performance of enzyme linked immunosorbent assay (ELISA) kit in detection of Hepatitis B virus(HBV) markers by using improved and optimized method ,so as to provide a practical and feasible method and reagents for clinical laboratory .Methods ELISA test was used for the detection of HBV markers .The gradient dilution method was used to e‐valuate the lower limit .The verifiation of cut off value was carried out based on clinical and laboratory standards institute (CLSI) EP12‐A2 document .Samples with cut off values were collected to evaluate the precision ,including repeatability and intermediate precision .The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000SR chemiluminescence instrument .Results The lower detection limit of HBsAg ,HBsAb , HBeAg ,HBeAb and HBcAb were 0 .2 IU/mL ,20 mIU/mL ,1 NCU/mL ,0 .75 NCU/mL and 0 .05 NCU/mL respectively .The cut‐off value(C50 )± 20% concentration included the concentration range between C5 and C95 .The within‐run coefficient of variation (CV)≤15% ,in sandwich method the between‐run CV≤25% ,in competition method the between‐run CV≤35% .The positive and negative coincidence rates in accuracy and comparing with i 4000SR all were more than 95% and all κ>0 .75 .Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs .

Abruptio Placentae ; complications ; Adult ; Disseminated Intravascular Coagulation ; diagnosis ; etiology ; therapy ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Prognosis ; Risk Factors

Adult ; Antiviral Agents ; therapeutic use ; Colonic Neoplasms ; diagnosis ; etiology ; therapy ; Cyclosporine ; adverse effects ; Esophageal Neoplasms ; diagnosis ; etiology ; therapy ; Female ; Ganciclovir ; therapeutic use ; Humans ; Immunosuppression ; adverse effects ; Immunosuppressive Agents ; adverse effects ; Liver Neoplasms ; diagnosis ; etiology ; therapy ; Liver Transplantation ; adverse effects ; Lymphoproliferative Disorders ; diagnosis ; etiology ; therapy ; Middle Aged ; Pharyngeal Neoplasms ; diagnosis ; etiology ; therapy ; Survival Analysis ; Tacrolimus ; adverse effects ; Time Factors

Spontaneous intracerebral hemorrhage (ICH) refers to primary non-traumatic parenchymal hemorrhage.Its mortality and disability are extremely high.A simple and easy clinical grading scale for ICH can not only evaluate the prognosis of patients,but also has an important guiding significance for clinical treatment and clinical research.This article reviews the contents of major intracerebral hemorrhage scales,external validation,advantages and disadvantages,and explains its scope of application and clinical application.

Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.

10.3969/j.issn.2095-4344.2013.23.007

Nitrite oxidoreductase(NXR) is the key enzyme responsible for the oxidation of NO2-to NO3-in nitrite-oxidizing bacteria.Since NXR is a dissoluble enzyme,located at the inner side of the membranes of cells,its function is dependent on the electron transfer chain related to membranes.This paper reviews the advances in study on NXR,including the structure,catalysis mechanism and the impact of different factors.New techniques applied in recent studies and research prospects are also presented.