Animals ; Danazol ; pharmacology ; Endometriosis ; drug therapy ; Endometrium ; drug effects ; Female ; Interleukin-6 ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

Objective To construct prokaryotic expression vector of hepatitis C virus NS5ATP1 gene, and to induce its expression in E. coli. To purify the fusion protein and obtain its polyclonal antibody from immunized New Zealand rabbits. Methods The NS5ATP1 gene, which was cut from the vector pGBKT7-NS5ATP1, which was self-constructed by the authors, was cloned into plasmid pET32a(+) to construct the pET32a(+)-NS5ATP1 prokaryotic expression vector. It was proved that the recombinant plasmid was constructed correctly by sequencing. And then the expression vector was transformed into the competent E. coli DH5? and BL21. After being induced with IPTG, the NS5ATP1 fusion protein was expressed and analyzed with SDS-PAGE and Western blot. The transformed bacteria were fragmented by ultrasonic and then separated by SDS-PAGE. The fusion protein formed inclusion body. They were then purified and re-natured through Ni+ affinity column chromatography. The purified pET32a(+)-NS5ATP1 fusion protein was used to immunize New Zealand rabbits to gain polyclonal antibody. The specificity and immunizing potence of polyclonal antibody were evaluated by Western blot and ELISA. Results After transferring pET32a(+)-NS5ATP1 plasmid into DH5? and BL21 and induced with IPTG, the NS5ATP1 fusion protein of about 56kD was highly expressed. SDS-PAGE analysis showed that the fusion protein products were mainly in inclusion body and expressed in the highest level at 4.5h of induction. The purified protein and polyclonal antibody were obtained successfully. ELISA manifested the titer of polyclonal antibody was over 1∶512 000. The high specificity was testified by Western blot. Conclusions The successful expression and purification of NS5ATP1 fusion protein and the preparation of NS5ATP1 specific polyclonal antibody will be valuable for the study on the biological function of NS5ATP1.

Objective To explore diagnosisand treatment of intestinal obstruction by foodballs in children. Methods The clinical datum of 21 cases of intestinal obstruction by foodballs in children were retrospectively ana- lyzed.Results While admission,only 2 cases could provide the history of having eaten foodballs wholly which could not be digested easily.The characteristics of clinical manifestation were intestinal obstruction completely or incom- pletely.All 21 cases had been treated conservatively first,only one case was successful and the other 20 cases had been cured by surgery afterwords.Conclusion There is no specific feature in clinical manifestations of intestinal ob- struction by foodballs in children,so it is difficult to make the diagnosis,but it must be considered suspicious especial- ly for those from rural area and with simple intestinal obstruction with reasons not known yet.If conservative therapy is failure in a short period,an operation should be done immediately.During operation,it is necessary to examine the whole digestive system thoroughly.

Objective To study the factors influencing the curative effect of one-stage bihteral total hip arthroplasty in a sequential procedure.Methods Thirty-six patients were indicated for one-stage bilateral total hip arthroplasty from February 1998 to March 2008.All cases were made with Smith-Peterson incision.Results The operative time was(3.2±1.2)h.The volume of blood transfusion during operation was (620 4±120)ml.All the 36 patients were followed up for(22.0±6.0)months.Except for 1 case of deep venous thrombosis of lower limbs,1 case of limb length discrepancy,there were no severe complications,such as postoperative infection,pulmonary embelism,dislocation,loosening or subsiding of components.Postoperative Harris scoreg of the joint function[(88.0±3.6)points]was significantly improved compared with their preoperative Harris scores[(28.0±4.2)points](P<0.05).Conclusions One-stage bilateral total hip arthroplasty is an effective method.However.the satisfactory effects can only be achieved by strict case selection,sufficient preoperative preparation,attention to operation procedures,prevention of complications and active recovery after operations.

Objective:To analyze the prognosis-related factors of ovarian serous adenocarcinoma in order to set up a prognostic model of serous adenocarcinoma and verify the effectiveness of the model as prognostic clinical criteria.Methods:The clinical, patholo-gical and follow-up data from 181 training samples with ovarian serous adenocarcinoma in Peking University First Hospital during January 1995 to December 2003 and another 42 detection samples with ovarian serous adenocarcinoma in Beijing Cancer Hospital during January 1999 to December 2005 were analyzed retrospectively. Kaplan-Meier univariate analysis was used to screen out prognostic factors; COX univariate and multivariate analyses were used to determine the risk coefficient of each factor and different layers in each factor; Pearson rank correlation analysis was used to identify the correlation of each factor. The prognostic model of ovarian serous adenocarcinoma was established on the conversion of risk coefficient to prognostic score and receiver operating characteristic (ROC) analysis was used to determine the cut-off value. The clinical data were collected to verify the sensitivity and specificity of the prognostic model based on the 3-year survival rate and Ki67 value.Results:The survival rate of patients with ovarian serous adenocarcinoma correlated with 6 factors including FIGO stage, histological grade, residual size, metastasis of lymph nodes, general condition after chemotherapy, and serum CA125 levels. The postoperative chemotherapy was an independent factor for prognosis. The prognostic model directly reflected the survival probability of classical COX risk ratio model, with higher score indicating lower survival probability and poorer prognosis. If combining prognostic score with Ki67 the sensitivity and specificity reached 64.7% and 96.0%, respectively.Conclusion:FIGO stage, histological grade, residual size, metastasis of lymph nodes, postoperative chemotherapy and serum CA125 were the prognostic factors of patients with ovarian serous adenocarcinoma. The prognostic model of ovarian serous adenocarcinoma can moderately reflect the actual survival status, and combination with Ki67 will increase the sensitivity and specificity.

Objective To discuss the clinical manifestations, diagnosis and treatment of solidpseudopapillary tumor of the pancreas (SPTP) in children. Methods Clinical data of 12 cases of SPTP admitted to Beijing Children's Hospital from Mar 2001 to Mar 2007 were collected and analyzed retrospectively. Result There were 7 females and 5 males among the patients. The mean age was 13.6 years. The main symptoms were abdominal pain and mass. Tumors located in the head in 4 cases, in the body and tail of the pancreas in 8 cases. Five cases underwent tumor enucleations, one case underwent body resection of the pancreas, two cases underwent Whippple's procedure including one recurrent case, five cases underwent body and tail resection of the pancreas plus splenectomy. All these 12 cases were followed up for 6 months to 6 years. One case lost follow-up 6 months after body and tail resection of the pancreas plus spleneetomy. Another one suffered from live tumor one year after operation and refused surgical exploration and lost follow-up. The other 10 cases were still alive. Conclusion SPTP are predominantly seen in adolescent females. For its benign clinical behavior and relatively good prognosis, surgical resection is the main choice of therapy.

Objective To investigate the role of Catbindin-D28k in the kidney on calcium metabolism.Methods VDR/CaBP-D28k double knockout(KO)mice was made.Body weight,diet intake and serum,urinary parameters and length,density of the long bones,histological staining of the tibia of WT,CaBP-D28(-/-),VDR(-/-)and VDR(-/-)/CaBP-D28k(-/-)mice were determined on regular and high Ca-Lac diet.Results On a regular diet,the double KO mice were growth-retarded more and smaller than VDR KO mice.Compared with VDR KO mice,the double KO mice had higher urinary calcium excretion and rachitic skeletal phenotype,which were manifested with higher serum parathyroid hormone levels,lower bone mineral density,and more distorted growth plate with mole osteoid formation in the trabecular region.On high calcium and high lactose diet,blood-ionized calcium levels were normal in both VDR KO and the double KO mice.However,in contrast to VDR KO mice,the skeletal abnormalities were not completely corrected in the double KO mice.Conclusion These results directly demonstrate that CaBP-D28k plays a critical role in maintaining calcium homeostasis and skeletal mineralization and suggest that its caleemic role can be mostly compensated by CaBP-D9k.

[Objective]To investigate the significance of the high-risk human papilloma virus (HPV)detection in the screening and diagnosis of cervical lesions.[Methods] The high-risk HPV DNA test results of 797 patients with cervical lesions who all accepted cytology and histopathology test were collected and analyzed retrospectively.[Results]The high-risk HPV DNA positive rates in cervicitis,cervical intraepithelial neoplasia(CIN)Ⅰ,CIN Ⅱ,CIN Ⅲ and cervical cancer were 53.41％(188/352),70.91％(117/165),87.63％(85/97),97.90％(140/143),97.50％(39/40),respectively.The sensitivity and negative predictive value of the high-risk HPV DNA detection for CIN Ⅱ and more serious lesions were 96.66％(318/329),93.29％(153/164),respectively.The detection rate of CIN Ⅱ and more serious lesions in patients with atypical squamous cells of undetermined significance(ASCUS)and positive high-risk HPV DNA was 30.03％(94/313),while the rate in patients with negative high-risk HPV DNA was 1.55％(2/129).[Conclusions] The more serious the cervical lesion is,the higher high-risk HPV DNA positive rate is.It is most closely related with CIN 11 and cervical cancer.The high-risk HPV DNA detection has high sensitivity and negative predictive value for CIN Ⅱ and more serious lesions.The high-risk HPV DNA detection has high negative predictive value in CIN Ⅱ and more serious lesions in ASCUS.

Objective:To study the influence of chemokine receptor blocker Met-RANTES upon endothelium adhesion and chemotaxis functions in response to T lymphocyte.Methods:In vitro,endothelium and T cell were mixed culture,which was used to observe endothelium adhesion and chemotaxis effect on T lymphocyte and expression of chemokine from endothelium,before and after using Met-RANTES.Results:When endothelium and T cell were mixed culture,endothelium expressed RANTES,MCP-1,MIP-1?,and the expression of RANTES was the largest.Met-RANTES could decrease endothelium adhesion and chemotaxis functions to T lymphocyte.Conclusion:Chemokines play an important role in endothelium adhesion and chemotaxis functions in response to T lymphocyte.Met-RANTES can decrease endothelium chemotaxis to T cell and then block endothelium firm adhesion with it.

? AlM: To observe the clinical curative effect of intravitreal Bevacizumab injection combined duplex technique in treatment of neovascular glaucoma ( NVG) .
?METHODS:Totally 25 eyes of 25 patients with NVG who underwent intravitreal Bevacizumab injection of 1. 0mg (0. 05mL), after the regression of iris neovascularization, 5 eyes with anterior chamber paracentesis fluid auxiliary controlled intraocular pressure. After 2wk, patients were treated by trabeculectomy and phacomulsification (9 eyes were implanted intraocular lens ) . The changes and complications of intraocular pressure, visual acuity, corneas and neovessels were observed after surgery, and followed up 12mo.
?RESULTS:After injection Bevacizumab in 25 eyes, iris neovascularization of 20 eyes subsided in 3 ~ 5d, and 5 eyes subsided in 7d. After controlling intraocular pressure, count of the corneal endothelial cell were 1 629±226mm2 , and none suffered decompensation of corneal endothelium after two- surgery of trabeculectomy and phacomulsification. After followed up 12mo, intraocular pressure of 20 eyes were controlled in normal range; 2 eyes could control in normal range after treated by a kind of anti-glaucoma medicine and 3 eyes was 34 ~38mmHg after treated by anti- glaucoma medicine. 9 eyes had improved vision after implanted intraocular lens.
?CONCLUSlON:lntravitreal Bevacizumab injection can subside iris and anterior chamber angle neovascularization effectively in a short time and reduce intraocular pressure. lt can also reduce the risk of bleeding during operation or after operation. lntravitreal Bevacizumab injection combined with two- surgery of trabeculectomy and phacomulsification can treat neovascular glaucoma effectively.