Objective:To study the anti-tumor effects by vaccination of mice with murine melanoma B16 cells expressing calreticulin (CRT) and Human papillomavirus 16 E2 fusion protein.Methods:Mice were inoculated with B16 cells expressing CRT,E2,or CRT-E2 by intraperitoneal injection. The inoculation was repeated after one week. The activity of CTLs and NK cells,the proliferation of T cells and the amount of IFN-? secreted by spleen lymphocytes were analysed.Results:The activity of CTLs and NK cells,and the amount of IFN-? secreted by spleen lmyphocytes from mice vaccinated with CRT-E2 fusion protein were significantly higher than those of the other groups(P

Objective To investigate the early period cesarean scars pregnancy diagnosis and its influence on prognosis. Methods Clincal data of 42 cases diagnosed as cesarean scar pregnancy from May 2006 to February 2011 treated in our hospital were retrospective analysed. Results All cases underwent B ultrasound examination,which showed gestational sacs or inhomogeneous echo-enclosed mass in the lower segment of anterior wall of uterus. Thirty-nine cases were successfully eonservatively treated,3 cases underwent cleanrance of focal lesion and neoplasty. Conclusion The cesarean scar pregnancy diagnosis depends on a B ultra-sound examination, and the color Doppler ultra sound is reliable for the diagnosis. Floss plants in the crater of scar and developed to uterus wall. Curettage guided by B ultra-sound after uterine artery embolization is a safe and efficient treatment method.

AIM: To detect the expression of the Calreticulin and HPV E2 Fusion Protein in B16, and study the effects on proliferation and apoptosis of B16 cell lines in vivo. METHODS: To construct eukaryotic fluoresce expression vector pEGFP-CRT-E2, pEGFP-CRT and pEGFP-E2. Then the recombinant plasmids were transfected into B16 cells by Lipofectamine 2000. The expression of proteins was detected by Western blot. The location of different GFP fusion proteins in B16 was tested by inverted fluoresce microscope. Flow cytometry was applied to detect the effects of fusion proteins on the growing of B16 and then the apoptosis effects of B16 induced by different proteins were observed. RESULTS: The correctly constructed recombinant plasmid pEGFP-CRT-E2 and the expression of CRT-E2 gene could be detected in B16 cells. Apoptosis of B16 cells could be detected after the transient transfection. Meanwhile, the apoptosis rate of B16 cells transfected by pEGFP-CRT-E2 was much higher than that of control cells. And cell cycle G1/G0 arrest was also found (P < 0.01). CONCLUSION: The eukaryotic expression plasmid pEGFP-CRT-E2 is successfully constructed and it could correctly express the fusion protein in B16 cells. And the B16 cells transfected by plasmid pEGFP-CRT-E2 could induce apoptosis.

Aim To explore the effect of Osthole (Ost)on the right ventricle remodeling in monocrot-alinetreated rats and the possible mechanism.Meth-ods ♂ SD rats were randomly divided into normal control group,model group,low dose of Ost treatment group (10 mg·kg -1 )and high dose of Ost treatment group (20 mg·kg -1 ).All rats were given a single dose of MCT 50 mg·kg -1 subcutaneously to establish the right ventricle remodeling except normal control group.Then the rats in Ost treatment group were ga-vaged once daily.After 28 days of administration,the right ventricle(RV)and left ventricle plus septum(LV+SEP)were weighed separately.RV hypertrophy in-dex (RVHI)were measured by the relative weight rati-o of RV to LV +SEP.The morphological changes of right ventricle were executed by HE staining.The pro-tein expression of PPARαand PPARγwere detected by Western blot.Results Compared with the control group,RVHI was increased obviously (P <0.05 ), myocardial hypertrophy and structure disorders were observed in model group.The protein expression of PPARαand PPARγwere down-regulated significantly in model group (P <0.05).Compared with the model group,the value of RVHI (P <0.05),myocardial hy-pertrophy,structure disorders were improved signifi-cantly in Ost treatment group.The protein expression of PPARαand PPARγwere up-regulated in Ost treat-ment group (P <0.05).Conclusion Ost can attenu-ate the right ventricle remodeling induced by MCT in rats,which may be related to up-regulating the expres-sion of PPARαand PPARγ.

Aim To investigate the effect of Icariin ( Ica) on remodeling of left ventricular in SHR and ex-plore the mechanism. Methods Twenty-one male SHR aged 14 weekswere randomly divided into model group(n=7), low-dose of Ica-treated group(20 mg· kg-1 . bid, n =7 ) , high-dose of Ica-treated group ( 40 mg·kg-1. bid,n=7), and WKY as control group(n=7 ) . Low- and high-dose of Ica-treated groups were given Ica from the age of 14 weeks to 26 weeks. The other rats in the model group and control group were given the same amount of double distilled water. Then, the content of hydroxyproline ( Hyp) was measured by ELISA. The morphological changes of the left ventricu-lar were observed by Masson staining. The mRNA and the protein expression of PPARα and PPARγ were ex-amined by real time RT-PCR and Western blot tech-nique respectively. Results Compared with the nor-mal control group, interstitium fibrosis and myofibrilla were lined up in disorder; the content of Hyp was in-creased and the mRNA and protein expression of PPARα and PPARγ were down-regulated in model group(P<0. 01). Compared with the model group,the myocardial cells were arranged less disorderly and the myocardial fibrosis was reduced; the content of Hyp was decreased in low-and high-dose of Ica-treated groups(P<0. 01 or P<0. 05);the mRNA and protein expression of PPARα and PPARγwere up-regulated in low- and high-dose of Ica-treated groups ( P <0. 01 or P<0. 05 ) . Conclusion Ica may attenuate left ven-tricular remodeling in SHR by up-regulating PPARαand PPARγ.

Objective To explore the effects of reoperation on treatment of recurrent pelvic endometriosis(RPEM).Methods The clinical data of 47 cases of RPEM reoperation in our hospital from April 2005 to October 2010 was investigated,and the efficacy was analyzed compared with the first operation data.Results The cases of painful nodules was significantly different between reoperation group and the first operation group(28 vs 14,x2 =8.436,P =0.004 ).There was significant difference on laparoscopic surgery cases between reoperation group and the first operation group (25 vs 40,x2 =7.259,P =0.007 ).The operation time in reoperation group was significantly longer than that in the first operation group( [ 106.4 ±41.0] min vs [ 78.4 ± 26.4 ] min,t=3.995,P ＜ 0.01 ),and the amout of intraoperative hemorrhage in reoperation group was more than that in the first operation group ( [ 143.2 ± 118.3 ] ml vs [ 70.6 ± 68.1 ] ml,t =3.660,P ＜ 0.01 ).However,there was no significant difference on symptoms,cyst location and clinical stage between these two groups(P ＞0.05).Conclusion Due to the pelvic adhesion would be dense and extensive in RPEM,it should be carefully dissected during reoperation.At the same time,the operator should pay attention to the anatomical location and try to restore the normal anatomy of the pelvic organs and physiological state,and try to reduce postoperative adhesions.Complete removal of the lesions is the key to improve the treatment effect and prevent recurrence and reoperation.

Aim To investigate the effect of icariin ( ICA) on renal interstitial fibrosis in SHR and explore its mechanism. Methods Fourteen male SHR of 13-week-old were randomly divided into model group( n=7 ) and ICA group ( n=7 ) , and WKY as control group (n=7). One week after adaptive breeding,the rats in the ICA group were given ICA 40 mg·kg-1,ig,bid to 26-week-old. The other rats in the model group and control group were given the same amount of normal sa-line. Then, the morphological changes of the kidney were observed by HE and Masson staining,respective-ly. The contents of plasma aldosterone andⅢcollagen were measured by double antibody sandwich method. The mRNA expressions of TGF-β1 , Smad2 , CTGF and FN were examined by real time RT-PCR. Results Compared with the normal control group, the kidney structures of model group were disordered,the mesang-ial matrix and the tubular interstitial fibrosis were in-creased. The contents of plasma aldosterone and Ⅲcollagen were increased in model group ( P <0. 01 ) . And the mRNA expressions of TGF-β1 , Smad2 , CTGF and FN in kidney tissues were up-regulated in model group( P <0. 01 or P <0. 05 ) . Compared with the model group,the kidney structures were improved and the contents of plasma aldosterone and Ⅲ collagen were reduced, as well as the mRNA expressions of TGF-β1,Smad2,CTGF and FN in kidney tissues were down-regulated(P<0. 01 or P<0. 05)in ICA group. Conclusion ICA may have anti-renal interstitial fi-brosis effect on SHR,and the mechanism might be re-lated to the reduced plasma aldosterone levels and the down-regulated expression of TGF-β1 and Smad2 .

Objective To investigate the effect of VvhA recombinant protein to the expression of IL-8 in human intestinal epithelial Caco-2 cells. Methods The VvhA recombinant protein(rVvhA) was ex-pressed by prokaryotic expression vector pET-28a (+)-whA in E. coli BL21 (DE3) , purified by Ni~(2+)-NTA affinity chromatography refoldod by stepwise deliquation together with dialysis methods and identified by Western blot. The cytotoxic of rVvhA to human Caco-2 cells was measured by CCK-8. The transcription of IL-8 mRNA in human Caco-2 cells induced by rVvhA was determined by RT-PCR, and the expression of IL-8 in human Caco-2 cells induced by rVvhA was determined by ELISA assay. Results rVvhA was purified with high purity up to 95%. The viability of human Caco-2 cells treated with 1.5 HU/ml rVvhA was inhibi-ted significantly (P < 0.05). The rVvhA can induce human Caco-2 cells to increase the transcription and expression of IL-8 in dose- and time-dependent manner. The transcription of IL-8 gene in human Caco-2 cells treated with 0.6 HU/ml rVvhA in 30 min can be up-regulated significantly, and the expression of IL-8 in human Caco-2 cells treated with rVvhA in 4 h can be increased significantly. Conclusion rVvhA has cy-totoxic to human Caco-2 and can increase the expression of IL-8, it might play a major role in the inflamma-tory reaction of rVvhA-exposed cells.

OBJECTIVE:To observe the effect of pathological lesion of renal tissue in rats with spontaneous hypertension (SHR),and study its mechanisms based on nuclear factorκB(NF-κB)signaling pathway. METHODS:21 SHR were randomly di-vided into model group and ICA low-dose,high-dose groups(20,40 mg/kg,denoted by ICA-L,ICA-H groups);other 7 homolo-gous Kyoto rats (WKY) were regarded as control group. All rats were intragastrically administrated,twice a day,for 11 weeks, rats in control group and model group received equal volume of double distilled water,ig. Pathological changes in renal tissue in each group were observed;Western blot method was used to detect protein expressions of p-NF-κB-p65,IκB and TNF-α in renal tissue. RESULTS:Compared with control group,model group showed disorder renal structure,narrow and irregular glomerular cysts;the protein expression of IκB was significantly down-regulated,protein expressions of p-NF-κB-p65 and TNF-α were signifi-cantly up-regulated(P<0.01). Compared with model group,the above-mentioned changes of rats showed improvement in ICA-L, ICA-H groups;the protein expression of IκB was significantly up-regulated in ICA-L,ICA-H groups (P<0.05 or P<0.01);the protein expressions of p-NF-κB-p65 and TNF-α were significantly down-regulated(P<0.01)in ICA-H groups;p-NF-κB-p65 pro-tein expression was significantly down-regulated in ICA-L group(P<0.05);while there was no significant difference in TNF-αpro-tein expression in ICA-L group(P>0.05). CONCLUSIONS:ICA plays a role in improving renal pathological lesion in SHR,and the mechanism may be related to inhibiting NF-κB signaling pathway.

Aims To observe the effect of Icariin( Ica) on apoptosis of renal tubular epithelial cell in spontane-ously hypertensive rats( SHR) , and to explore its pos-sible mechanism. Methods 21 male SHR of 14 weeks were randomly divided into model groups, the Ica low and high dose groups (20 or 40 mg·kg-1 ,ig, bid, to 26 weeks ) , 14 week-old male homologous Kyoto rats ( WKY ) as control group, and the number of each group was 7 . WKY group and model group were ad-ministered by gavage with the same volume of double distilled water. The pathological changes were observed by using the HE staining and the apoptosis was detec-ted by TUNEL method in renal tubular epithelial cell, respectively. The mRNA levels of Bok, Bax, Bcl-2 were detected by the real time RT-PCR and the protein expressions of Bcl-2 , Bax and Active caspase-3 were detected by Western blot method in the renal tissue. Results Compared with WKY, renal capsule was nar-row and irregular, and glomerular mesangial matrix was increased with the cell arrangement disorder and the capillary dilation and congestion. Several glomeruli shrank and renal tubular epithelial cell was edema with luminal stenosis in model group. The apoptosis of renal tubular epithelial cell was evident and the mRNA levels of Bok and Bax, as well as the protein expressions of Bax and Active caspase-3 were significantly up-regula-ted in model group, while the mRNA Level and protein expression of Bcl-2 significantly down-regulated ( P <0. 05 or P<0. 01). Compared with model group, renal glomerular capsule widened, proliferation of glomerular mesangial matrix was reduced, and cell arrangement disorder and capillary dilation and renal tubular lumen stenosis were improved in Ica group. Renal tubular ep-ithelial cell apoptosis was decreased evidently, and the mRNA level of Bax, Bok and protein expression of Bax were significantly down-regulated in Ica group, and the mRNA level and protein expression of Bcl-2 in Ica-H group were significantly up-regulated while the protein expression of Active caspase-3 significantly down-regu-lated( P <0. 05 or P <0. 01 ) . Conclusion Ica can inhibit the apoptosis of renal tubular epithelial cell in spontaneously hypertensive rats and its mechanism may be related to the down-regulation of Bok, Bax, and Active caspase-3 , accompanied with the up-regulation of Bcl-2 .