Objective:The envelope protein Domain I(DI) of type II mutated dengue virus from a DHF patient (B-E) was expressed,refolded and purified.Its immunogenicity was also identified.Methods:The 1-476 bp of B-E was cloned into pET28a(+),and it was transformed into E.coli after digestion and sequencing.The inclusion body was denatured and refolded after inducing with IPTG.Polyclone antibody was obtained by immunizing C57BL/6 and BALB/c mice with purified B-E,and identified with Western blot and ELISA.Results:The cDNA of B-E gene was inserted into pET28a(+) vector and transformed into Rosetta(DE3).The protein of B-E was mainly in inclusion body and the molecular weight was 20 kD as predicted.Western blot was used to identify the recommend protein with anti-his antibody.The soluble recombination protein was purified after denaturation and refolding.The polyclonal antibody was obtained by immunizing BALB/c and C57BL/6 mice and could be used in Western blot and ELISA assay.The titer of polyclonal antibody from C57BL/6 was 1:12 800 through ELISA assay,and titer of polyclonal antibody from BALB/c was 1:500 through Western blot.Conclusion:All the results suggests that B-E is immunogenic in BALB/c and C57BL/6 mice.

Objective:BALB/C mice model infected with different Dengue Type 2 virus (DV 2) strains was evaluated and dynamic patterns of humoral immune responses caused by the various viral strains were investigated.Methods:The BALB/C mice were multisubcutaneouly injected by different DV 2 strains. The dynamic changes of the anti-DV 2 IgM/IgG antibody were observed in the infected BALB/C mice by the indrect ELISA and the viremia was observed.Results:The productions of specific antibodies in the infected BALB/C mice after primary and secondary injection with different DV 2 strains showed some difference. Howerer, BALB/C mice infected by the DV 2 B strain from a DHF patient showed the prolonged viremia.Conclusion:The dynamic curve of specific antibody and the viremia induced by various DV 2 strain (DV 2 NGC strain.V.DV 2 B strain) were different in some existence.

Objective:BALB/C mice model infected with DV2 NGC strain was evaluated and dynamic patterns of cytokines were investigated.Methods:The dynamic changes of cytokines were observed in the infected BALB/C mice model by indirect ELISA was observed.Results:The dynamic levels of IL-4、IFN-? in the plasma of BALB/C mice after primary and secondary infected DV2 NGC strain were different.The results showed that the levels of IL-4 were significantly increasing in the early primary infected BALB/C mice while that of IFN-? were lower;In secondary infection ,IL-4 reached a peak on the first day( 4294 668 ? 349 038 pg/ml) and then the levels of IFN-? were higher on the forth and eleventh day.The concentrations of the cytokines produced infectious animals had closely relationship with the doses of DV2.Conclusion:Infection eliciting a dominant humonral immune response induced a high expression of Th2-related cytokines,whereas those show some appearance of Th1 cytokines.Th1 response,as described late for the disease infected with DV2 NGC strain,played a role while it inhibited Th2 response to some extend.Thus,it was responsible to recovery.Th cell and the cytokines were critic to the DV infective diseases.

Objective To discuss the effect of von willebrand factor (vWF) antibody on idiopathic thrombotic thrombocytopenic purpura(TTP) .Methods 28 cases of idiopathic thrombotic thrombocytopenic purpura were selected in our hospital from January 2011 to June 2013 ,and 30 healthy persons as control group ,ADAMTS13 and vWF of two groups were detected .Results 3 cases of patients were detected with positive vWF antibody ,vWF antibody negative in the remaining patients .3 patients with positive vWF antibody ADAMTS13 antibodies were negative ,ADAMTS13 levels were lower than the normal value .The levels of ADAMTS13 in vWF antibody positive patients was significantly lower than that of vWF antibody negative patients and the control group ,(P<0 .01) .vWF antibody positive patients plasma vWF antibody A was higher than vWF antibody negative and the control group (P<0 .01) .In idiopathic TTP after PE ADAMTS13 antigen increased significantly ,vWF antibody and A levels decreased significantly (P<0 .05) .Conclusion vWF antibodies in idiopathic thrombotic thrombocytopenic plays an important role in the pathogenesis of purpura ,vWF antibody may affect patients ADAMTS13 ,promote vWF complex formation ,effect of disease .

Objective:To observe the effect of different concentration HMGB1 on the secretion of TNF-αand NO from Ana-1 infected with DEN2 and virus copy.Methods:DEN2 were proliferated and identified by conventional methods.The adherence of DEN2 to Ana-1 was observed by direct immunofluorescence and RT-PCR.The level of virus mRNA were detected by qRT-PCR.The concentration of TNF-αwas detected by ELISA.The concentration of NO was detected with Griess reagent.Results:Ana-1 was able to adhered for DEN2.Compared with DEN group,the inhibition ratio(%) of the level of virus mRNA in D-HMGB1-1 group,D-HMGB1-10 group,D-HMGB1-100 group,D-HMGB1-1000 group was 41.53 ±2.12,55.30 ±1.59,74.75 ±1.12,86.35 ±1.42.Compared with DEN group,the level of TNF-αand NO decreased in D-HMGB1 groups(P<0.05).Conclusion:HMGB1 can be effectively regulated of Ana-1 secreted inflammation factor of infected with DEN2,and inhibited DEN2 replication.

Echinococcosis is a zoonotic epidemic parasitic disease, which is caused by the infection with larvae of Echinococcus in humans, and the disease is also known as hydatid disease. The liver and lung are the most vulnerable organs. Aspergillus is a saprophytic fungus. Aspergillus infection is common in patients with immune deficiency. Pulmonary echinococcosis complicated with Aspergillus infection is very rare in patients with normal immune function. It is a case report in the literature at home and abroad. A case of bilateral isolated pulmonary echinococcosis complicated with Aspergillus infection with normal immune function and no other accompanying diseases admitted to Lanzhou University Second Hospital is reported as follows, and literature review is conducted in combination with Chinese and foreign literatures, so as to provide some reference value for diagnosis and treatment of this kind of disease.

Objective To investigate the effect of comfort nursing based on teamwork on patients after laparoscopic myomectomy of hysteromyoma .Methods One hundred and fifty-six patients who chose laparoscopic myomectomy of hysteromyoma were randomly divided in the intervention group and the control group, each with 78 cases.The control group received conventional nursing while the intervention group received comfort nursing including preoperative interactive supervision , intranperative comfort nursing and postoperative visits.Effect of nursing was compared between two groups .Results The operation time , called the eye opening after the operation , called the open fist time and the tracheal extubation time was respectively ( 52.63 ±7.25 ) min, (18.62 ±1.65)min, (22.15 ±1.32)min and (25.37 ±1.52)min in the intervention group, shorter than (58.72 ±7.31)min,(23.05 ±1.72)min,(25.67 ±1.38)min and(29.62 ±1.56)min in the control group, and the differences were statistically significant ( t=5.224 ,16.415 ,16.279 ,17.233 , respectively;P<0.01).The pain score 12, 24, 48 and 72 h after operation was respectively (4.32 ±1.42),(3.29 ±1.27), (2.37 ±1.05),(1.25 ±1.02)in the intervention group, lower than(5.47 ±1.52),(4.56 ±1.31),(3.56 ± 1.08),(1.82 ±1.05)in the control group, and the differences were statistically significant (t=4.883,6.147, 6.977 ,3.439 , respectively; P <0.05 ) .The time of first being out of bed was ( 12.35 ±3.17 ) h in the intervention group and ( 17.52 ±3.23 ) h in the control group , with statistically significant difference ( t =10.089,P<0.05).Sleep time 24, 48 and 72 h after surgery as well as hospitalization time was respectively (5.72 ±2.31)h,(6.55 ±1.58)h,(7.87 ±1.25)h and (7.13 ±1.28)d in the intervention group, better than (17.52 ±3.23)h,(3.59 ±2.35)h,(5.37 ±1.63)h,(6.58 ±1.27)h and(8.67 ±1.59)d in the control group, and the differences were statistically significant ( t =5.709,4.591,6.393,6.663, respectively; P <0.01).The incidence of complication was 6.41%in the intervention group, and 24.36%in the control group, with statistically significant difference (χ2 =9.652 ,P<0.05 ) .Conclusions Comfort nursing for patients after laparoscopic myomectomy of hysteromyoma can improve their surgical compliance , shorten operation time and awakening time , reduce postoperative complications and enhance surgical effect .