Objective:To prepare and identify a polyclonal antibody againstadam28 gene product.Methods:Theprotein coding region of ADAM28 was amplified by RT-PCR and cloned into pMD18-T Vector to produce the newconstruct, pMD18-T-adam28. The cloned ADAM28 segment was cut with two restriction enzymes and theadam28fregment was directed into the prokaryotic expression vector, pGEX-4T-1,to produce the expression vector pGEX-4T-adam28. The recombinant plasmid was transformed intoE. coliDH5?and GST-ADAM28 fusion protein was ob-tained after the inducement by IPTG. The fusion protein was extracted and purified by SDS-PAGE,and the newpro-tein band of 35 300 was isolated as antigen, the antigen was injected into rabbits to produce polyclonal antibody a-gainst ADAM28 product.Results:The expression vector pGEX-4T-adam28 was constructed successfully,and GST-ADAM28 fusion protein was obtained. The rabbit serum containing polyclonal antibody against ADAM28 productwas obtained and the antibody was purified by salting out method. Western blot analysis displayed that the antibodyhad high specificity. ELISA analysis confirmed that the titer for the antibody reached 1∶16 000.Conclusion:Thepolyclonal antibody against ADAM28 product with high titer is successfully prepared,it may be used for further studyof the role and expression of ADAM28 during tooth development.

Objective:To investigate spatiotemporal expression of ADAM28 in mouse tooth germ development.Methods:Immunohistochemistry and image analysis technique were used to observe the expressions of ADAM28 at mouse tooth germ development stages.Results:Different expression levels of ADAM28 at tooth germ development stages were observed.At cap stage,ADAM28 was found strongly positive in oral epithelial,stellate reticulum cells of enamel organ,basement membrane,dental papilla cells and dental sac cells.At late bell stage,positive staining was found in ameloblasts,enamel matrix,epithelial root sheath and dental papilla cells.At crown and root development stage,positive staining for ADAM28 was detected in ameloblasts,odontoblasts,cementoblasts,epithelial root sheath,dental papilla cells and dental sac cells.Conclusion:ADAM28 participates in crown and root morphogenesis process ranging from bud stage to late bell stage and from matrix secretion to sclerous tissue formation.It might play an important role in early formation,proliferation and differentiation of odontogenic mesenchymal cells.

Objective:To investigate the temporal and spatial distrib ut ions of collagenⅠ and Ⅲ during mouse tooth germ development and their function s during tooth mineralization.Methods:Immunohistochemistry stain ing technique was used to test the expressions of collagen Ⅰ and Ⅲ during mous e tooth germ development. Results:collagen Ⅲ was positive in or al epithelial cells in bud stage,in oral epithelial cells and in stellate reticu lum cells in cap stage. During bell and differentiation stage,collagen Ⅲ was positive in oral epithelial cells, stellate reticulum cells, dental papilla cell s and dental sac cells. During P2-10 d(crown development stage), collagen Ⅲ was expressed possitively in ameloblasts,enamel matrix,odontoblasts,predentin, dental papilla cells,dental sac cells and pulp tissues. During P10-30 d(toot h root development stage),collagen Ⅲ was strongly positive in Hertwig's epithe lial root sheath, cementum, alveolar bone and periodontal ligament cells apart f rom above mentioned cell types. CollagenⅠ was not expressed in bud stage and wa s positive in oral epithelial cells,stellate reticulum cells in cap stage. Durin g bell and differentiation stage,collagen Ⅰ was positive in oral epithelial ce lls, stellate reticulum cells, dental papilla cells and dental sac cells.After P2 d (crown and root development stage), the distribution and expression of co llagen Ⅰ were similar to those of collagen Ⅲ.Conclusions:Coll agenⅠand Ⅲ are involved in tooth germ and tooth tissue development. But the fu nction of collagenⅢ is more extensive than that of collagenⅠ.

Objective To evaluate the etiology, clinicopathological characteristics, treatment and prognosis of VIN Ⅲ. Methods Thirty-five patients with VIN Ⅲ admitted in Cancer Hospital of Chinese Academy of Medical Sciences between 1993 and 2008 were analyzed retrospectively. Results All 35 patients attended to hospital with the symptoms of pruritus vulvae and lumps. Among of them, there were 28 (80%) cases whose lesions presented as multiple plaques, while 7 (20%) eases lesion were monofocal.Fifteen out of 35 eases had coexistent with human papillomavirus (HPV) related lesions, and there were the evidence of HPV in 22 patients whose pathologic sections revealed koilocytes. 34 cases (97%) except one received different types of surgical resections, including wide local resection (14/34), simple vulvectomy (16/34) and simple vulvectomy plus perianal skin resection (4/34), while there two cases shown positive cut-edges and one of them received radiotherapy postoperatively. Four out of the 34 cases recurred locally in 2, 4, 6 and 22 months and received surgical treatment again or laser therapy, respectively. The median follow-up period was 66 months ( range 1-166), and none of them were died of the disease itself except one case died of the concurrent cervical cancer. Among 26 cases received the investigation of the quality of life,there were nine cases (35%) suffered from sexual dysfunction after the operation, and one of them received vulvoplasty. Conclusions VIN Ⅲ has good prognosis. The development of VIN Ⅲ may be related to the infection of HPV, because most of them concurrent with HPV diseases. Its primary treatment is surgical resection, while affecting on the quality of life.

This paper presents some ideas and practices to realize the homogeneity quality management process within the Beijing Children Hospital Group,the largest specialized medical group in the country.This research aims at providing references and thoughts on homogeneity quality management for trans-regional medical unions in the country.

Objective To compare the differences in 4 sets of gross tumor volumes (GTVs) generated from FDG PET/CT in cervical cancer,and determine the optimal method for target volume delineation of cervical cancer.Methods Sixteen cervical cancer patients with 28 primary or metastatic lesions underwent FDG PET/CT.CT and PET images were coregistered,and transferred to Pinnacle therapy planning workstation.Four sets of GTVs were defined.The first set ( GTVvis ) was manually contoured using a visual method on PET images.The second set ( GTV40 ) was autocontoured using a threshold of 40％ of the maximum intensity level for PET images.The third set ( GTV2.5 ) used an autocontour of standardized uptake value (SUV) of 2.5 around the tumor.By phantom measurements we determined an algorithm ( threshold =(mean target concentration + 2.623)/1.975),GTVfunction was defined using this method as the fourth approach.The volumes of 4 sets of GTVs were compared with group t-test.ResultsThe average volumes of GTVvis,GTV2.5,GTV40,and GTVfunction were 63.41,53.20,41.33,and 61.84 cm3. There was no significant difference between GTVfunctoon,GTV2.5 and GTVvis ( t =1.05,0.91,P =0.305,0.37 ),but GTV40were smaller than GTVvis.The SUVmax and target to background value had no significant influence on the differences between GTV40 and GTVfunction or GTVvis ( t =0.00,- 0.34,0.92,0.35,P =1.000,0.746,0.374,0.737),but they had significant influence on the difference between GTV2.5 and GTVvis (t =- 3.87,3.16,P =0.002,0.016).ConclusionsGTVvis,GTV2.5 and GTVfunction all could be used for target delineation if the method can define the GTV.The difference between the GTVfunction and GTVvis was the smallest;GTV40 was smaller than GTVvis. GTV2.5 was significantly influenced by SUVmax and target to background value of the legions.

In recent years, antiangiogenic drugs based on VEGF and VEGFRs signaling pathway have been widely used in the treatment of malignant tumors. Apatinib is an orally bioavailable small-molecule antiangiogenic agent and can specifically inhibit the tyrosine kinase activity of VEGFR, thereby inhibiting tumor angiogenesis. Apatinib is the first-level recommendation of third-line treatment of gastric cancer in CSCO Guidelines for the Diagnosis and Treatment of Gastric Cancer published in 2018. Apatinib has been proved to be effective and safe in gastric, lung and breast cancers. In addition, the drug shows great potential in the treatment of a variety of solid tumors. This article reviews the recent progress on the mechanism, clinical efficacy, related efficacy predictors of apatinib and its combination with other antitumor drugs.

Objective To predict clinical chemotherapy sensitivity of primary ovarian cancer by jointing adenosine triphosphate(ATP) - tumor chemo-sensitivity assay(TCA) method in vitro and detection of drug resistance genes, provide reference for clinical treatment. Methods Forty-seven primary epithelial ovarian tumor samples were collected from the patients who received cytoreductive surgery. Viable ovarian cancer cells obtained from malignant tissue were tested for their sensitivity to carboplatin (CBP), cisplatin (DDP), paclitaxel(PTX) and CBP + PTX using ATP-TCA method in vitro; at same time, real-time quantitative PCR was used to analysis BRCA1 and ERCC1 mRNA relative expression in forty-six specimens (1 frozen tumor samples mRNA were not detected due to serious degradation). The relationship between ATP-TCA test results, clinical indicators, and the effectiveness of the joint prediction on clinical chemosensitivity by combining these two methods were statistically analyzed using chi-square test. Results (1)The results showns that three programs of DDP,CBP and PTX + CBP were significantly related with clinical results(P<0.05) in vitro, in which the compliance rate in PTX + CBP program was the highest 83%(39/47) ,and the predictive sensitivity, predictive specificity, positive predictive value, negative predictive value and predictive accurate rate were 90%,71%,84% and 80% ,respectively.PTX + CBP combined in vitro test results was also related with residual tumor size and neoadjuvant chemotherapy, which was more prone to drug resistance with residual tumor larger than 2 cm (P = 0. 023) and with neoadjuvant chemotherapy (P = 0.011). (2) BRCA1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was 0.673 ± 2.143 and - 1.436 ± 2.594 (P=0.008), ERCC1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was -0.529 ± 1.982 and - 3.188 ±2.601 (P =0.001). There were also significant correlation among the expression levels of BRCA1 ,ERCC1 mRNA and clinical efficacy (P<0.01). (3)ATP-TCA and detection of drug resistance genes combined to predict the clinical application of PTX + CBP resistance may occur in 8/9 cases. Conclusions ATP-TCA may be an ideal method of in vitro drug sensitivity testing method, which could effectively predict clinical chemotherapy sensitivity. Combination of the drug-resistant associated genes detection method and the ATP-TCA method can increase the predictive effectiveness of ovarian cancer chemosensitivity and guide individual chemotherapy of ovarian cancer.

OBJECTIVESTo investigate the risk factors for cervical cancer in the areas of high incidence, and provide evidence for current intervention of cervical cancer.

METHODSIn the areas of Xiangyuan County, Shanxi Provicne with high incidence of cervical cancer, 1 997 women were interviewed using a questionnaire, including baseline information, menstrual, marital and pregnancy histories, sexual behavior, health habits, contraception, medical history and family history of cancer, etc., after its screening with six kinds of methods. All subjects, including 84 cases with pathological diagnosis of greater than cINI, and 1 784 cases with pathological diagnosis of normal, were tested for high-risk HPV.

RESULTSThe overall rates of HPV infection were 20.8% (415/1 997) in high-risk subjects, 97.7% and 14.2% in the cases and control groups, respectively. Univariate analysis showed that risk factors with statistical significance included high-risk HPV infection, age at first sexual intercourse, history of pregnancy and abortion, the number of sexual partners and family history of cancer. Analysis with non-conditional logistic regression model revealed high-risk HPV infection, multiple sexual partners and family history of cancer associated obviously with occurrence of cervical cancer. In addition, there was significantly positive relationship between HPV infection, which increased with the number of sexual partners, and extramarital sexual activity both in males or females.

CONCLUSIONSThe main risk factor for cervical cancer in this region was high-risk HPV infection, which related to sexual behavior, hygienic habits during menstruation and puerperium. It was particularly important to detect and treat precancerous lesions and to implement behavior modification. In addition, further research on genetic susceptibility was suggested.

Abortion, Induced ; Adult ; Analysis of Variance ; China ; epidemiology ; Female ; Humans ; Interviews as Topic ; Multivariate Analysis ; Papillomaviridae ; Papillomavirus Infections ; epidemiology ; Risk Factors ; Sexual Behavior ; Sexual Partners ; Surveys and Questionnaires ; Tumor Virus Infections ; epidemiology ; Uterine Cervical Neoplasms ; epidemiology ; virology

Objective To investigate the expression of soluble vascular endothelial growth factor receptor 1 (sFlt1) in human umbilical cord-derived mesenchymal stem cells (HUCMSC) under the condition of inflammatory cytokine and co-culture with rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs).Methods ① HUCMSC were cultured alone or stimulated by Tumour necrosis factor (TNF)-αt,interferon (IFN)-γor the combination of TNF-α and IFN-γ.The sFh1 levels of each group were detected by enzyme-linked immunosorbent assay (ELISA) at 24 h,48 h and 72 h respectively.② RA-FLSs and HUCMSC were cocultured in a transwell system with both of them cultured alone as control for 72 hours.ELISA was used to detect sFlt1 levels in each group.T test was used for comparison between two groups,and ANOVA was used to compare multi-group variables.Results ① The sFlt1 levels of TNF-α + IFN-γ group were significantly higher than those of the control group at 24 h and 48 h (24 h (5.4±0.4) ng/ml vs (2.8±1.7) ng/ml,t=0.942,P=0.026;48 h (7.2±0.8) vs (4.3±1.0) ng/ml,t=4.285,P=0.005),while that at 72 h was not significantly different [(9.1±1.7) ng/ml vs (7.0±1.4) ng/ml,t=0.683,P=0.52].And no significant difference in sFlt1 levels between control group and TNF-α group or IFN-γ stimulation group were observed.② Compared with RA-FLSs,the expression of sFlt1 in HUCMSC was significantly higher (8.19±3.64) ng/ml vs (0.19±0.08) ng/ml,t=7.280,P＜0.01].After co-cultured with RA-FLSs and HUCMSC,the sFlt1 concentration of the co-culture group was significantly higher than that of HUCMSC group [(17.26±6.92) ng/ml vs (8.19±3.64) ng/ml,t=3.985,P=0.000 7].Conclusion The sFlt1 expression of HUCMSC is up-regulated by inflammatory cytokine TNF-α combined with IFN-γ.RA FLSs may promote the co-cultured HUCMSC to increase sFlt1 expression by secreting inflammatory factors.