Objective　To investigate the expression of estrogen receptor β (ERβ ) mRNA and its significance in endometrial carcinoma. Methods　Thirty-six normal and 45 cancer tissue samples of endometrium were obtained, and the RNA was isolated from the tissues by the acid guanidium thiocyanate-phenol-chloroform extraction method. Reverse-transcription polymerase chain reaction (RT-PCR) was used to amplify the ERβ exon 5 wild type mRNA (ERβ WT) and splicing variant mRNA (ERβ E5SV). Electrophoresis of polymerase chain reaction product was performed and the ratio of ERβ E5SV/ERβ WT was calculated. The sequencing of ERβ E5SV and ERβ WT was carried out by express DNA sequencer. Results　ERβ E5SV and ERβ WT were both expressed in normal endometrium and endometrial cancer tissues. A region of 139 bp from base pair 812 to 950 (the hormone-binding domain) was found to be deleted in ERβ E5SV sequence as compared to ERβ WT. The quantity of ERβ E5SV was more than that of ERβ WT in every normal endometrium, and was less in every endometrial cancer, the ratio of ERβ E5SV/ERβ WT was 2.47±0.99 in normal endometrium, and 0.55±0.12 in endometrial cancer (P<0.000 1). The ratio of ERβ E5SV/ERβ WT decreased with the grade from 1 to 3 (P<0.01). Conclusion　Overexpression of ERβ WT relative to ERβ E5SV may correlate with the genesis of endometrial carcinoma.

Objective To investigate the expression of estrogen receptor ? (ER? ) mRNA and its significance in endometrial carcinoma. Methods Thirty six normal and 45 cancer tissue samples of endometrium were obtained, and the RNA was isolated from the tissues by the acid guanidium thiocyanate phenol chloroform extraction method. Reverse transcription polymerase chain reaction (RT PCR) was used to amplify the ER? exon 5 wild type mRNA (ER? WT) and splicing variant mRNA (ER? E5SV). Electrophoresis of polymerase chain reaction product was performed and the ratio of ER? E5SV/ER? WT was calculated. The sequencing of ER? E5SV and ER? WT was carried out by express DNA sequencer. Results ER? E5SV and ER? WT were both expressed in normal endometrium and endometrial cancer tissues. A region of 139 bp from base pair 812 to 950 (the hormone binding domain) was found to be deleted in ER? E5SV sequence as compared to ER? WT. The quantity of ER? E5SV was more than that of ER? WT in every normal endometrium, and was less in every endometrial cancer, the ratio of ER? E5SV/ER? WT was 2 47?0 99 in normal endometrium, and 0 55?0 12 in endometrial cancer ( P

ObjectiveTo study the associated factors with pelvic lymph node metastasis of endometrial carcinoma and the effect of pelvic lymphadenectomy on prognosis of the disease MethodsTotally 102 patients with endometrial carcinoma who underwent pelvic lymphadenectomy (90 patients) or lymph node biopsy (12 patients) in our hospital from Jan 1981 to Dec 2002 were recruited The relationship between various clinicopathologic factors and pelvic lymph node metastasis was analyzed Prognosis of ninety patients with pelvic lymphadenectomy was compared with 90 patients without pelvic lymphadenectomy (control group) in the same period The 5-year survival was calculated by life table method Results The incidence of pelvic lymph node metastasis increased in patients with low grade(46%),deep myometrium invasion(42%), cervical involvement(44%), positive peritoneal cytology(52%), adenexal metastasis(75%) and distant spread(100%) The 5-year survival was lower in patients with lymph node metastasis (37%) than that without lymph node metastasis (89%, P

Objective To study the expression and clinical significance of Notch3 and Notch intracellular domain (NICD) in ovarian carcinoma and the effects of N-[N-(3 ,5-difluorophenyl) acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis in OVCAR3, A2780 ovarian carcinoma cell lines. Methods Western blot was used to detect the expression of NICD in the tissues from 58 ovarian carcinomas patients and 21 normal ovarie, who were admitted in Peking University First Hospital from July 2006 to June 2009. Immunohistochemistry was also used to detect the expression of Notch3 in these tissues. The relationship with clinical features of ovarian carcinoma was also analyzed. Proliferation of OVCAR3 and A2780 ovarian cancer cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in OVCAR3 and A2780 cells incubated with DAPT was detected by western blot. Results (1)The expression level of NICD in ovarian carcinomas was significantly higher than that in normal ovarian tissues (1.64 ±0. 19 vs. 0.98 ±0.20;P <0.05). The NICD expression was higher in ovarian cancers with low grade or advanced stage than those in high-middle grade or early stage,respectively (1.90 ± 0. 22 vs. 1.25 ± 0. 21,1.80 ± 0. 21 vs. 1.21 ± 0. 15; all P < 0. 05). The Notch3 protein was stained positively in cytoplasm, nuclear and cell membrane. The expression of Notch3 was higher in ovarian carcinomas than that in normal ovaries [78% (45/58) vs. 24% (5/21); P < 0. 01]. While,there were no stasistical difference in different pathological types, stages, differentiation of ovarian carcinoma. There was no difference between the patients with adjuvant chemotherapy or not. (2)After OVCAR3 and A2780 cells incubated with DAPT 24, 48, 72 hours, NICD expression was significantly lower than that in control group (P < 0. 05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells were depended on the concentrations and times. Conclusions Notch3 and NICD may play a key role in the occurrence and progress of ovarian carcinoma. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of OVCAR3 and A2780 cells may be due to decreased the formation of NICD.

Objective To study the expression of Notch 1,Jagged1 and Notch intracellular domain (NICD) in epithelial ovarian carcinoma tissues and analyze the clinical significance.To explore the activity of γ-secretase in epithelial ovarian carcinoma cell line SKOV3 and the effect of N-[N-(3,5-dil uorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT),a γ-secretase inhibitor on the activity of γ-secretase in SKOV3.Methods Immunohistochemistry staining method was performed in 43 patients with epithelial ovarian carcinoma and 11 patients with benign epithelial ovarian tumor to detect the expression of Notch1,Jagged1 and NICD.The differences of expressionof Notch1,Jagged1 and NICD between malignant and benign ovarian tumors was compared and alsoanalyzed the correlation with clinicopathological parameters of ovarian carcinoma.Human serous ovarian cancer cell line SKOV3 and immortalized nontumorigenic ovarian epithelial cell line T29 were incubated in vitro.The activities of γ-secretase in SKOV3 and T29 with dimethyl sulfoxide (DMSO) and DAPT were detected respectively by Gal4VP16/UAS and dual luciferase reporter assay system.Results (1) The immunohistochemical composite scores (ICS) of Notch1 in epithelial ovarian carcinoma (6.7±2.2) were not significantly different with those in benign epithelial ovarian tumor (5.4± 2.7,P=0.153),while the ICS of Jagged 1 and NICD in epithelial ovarian carcinoma (5.3± 2.4,5.3± 2.3) were higher than those in benign epithelial ovarian tumor (1.6± 1.4,3.1± 1.7; all P＜0.01).The expression of Notch 1,Jagged 1 and N ICD had no correlation with patients' aged,history of carcinoma,ascites,the level of serum CA125,maximum length of ovarian tumor,Federation International of Gynecology and Obstetrics (FIGO) stage,grade and pathology subtypes (all P＞0.05).The hazard ratio between the high expression of Notch1,Jagged1,or NICD and the moderate to low expression of Notch1,Jagged1,or NICD,and Jagged1 were 0.771,1.648 and 1.316,respectively (all P＞0.05).The 5-year survival rate and median survival time between the high expression of Notch,Jagged 1 or NICD in subgroup and moderate to low expression in subgroup were of no difference (all P＞0.05).The activity of γ-secretase in SKOV3 was significantly higher than that in T29 [(12.2± 1.4)％,P=0.019].(2)After DAPT treated,the relative activity of γ-secretase in SKOV3 (50 μmol/L) was declined from (100.0±5.3)％ to (6.6±0.8)％ (P=0.001).Conclusions Jagged1 and NICD in Notch1 pathway may play a key role in the occurrence of ovarian carcinoma.The activity of γ-secretase in epithelial ovarian carcinoma was higher than that in ovarian epithelial cell which suggest that DAPT,γ-secretase inhibitor,may become the target of ovarian carcinoma treatment.

Objective To study the expression and clinical significance of Notch intracellular domain (NICD) in cervical cancer and the effects of N-[N-(3,5-difluorophenyl)acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a γ-secretase inhibitor on the proliferation and apoptosis of cervical cancer cell lines. Methods Western blot was used to detect the expression of NICD in the tissues of 40 cervical cancers and 21 normal cervix and its relationship with clinical features of cervical cancer was also analyzed. Proliferation of SiHa and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in SiHa and HeLa cells incubated with DAPT was detected by western blot. Results The expression level of NICD in cervical cancers was significantly higher than that of normal cervical tissues (1.237±0.353 vs 0.938±0.105, P<0.05). The NICD expression was higher in cervical cancers with high grade,lymph node involvement and parametrial invasion than that with low-middle grade (1.496±0.540 vs 1.150±0.216), without lymph node involvement (1.419±0.532 vs 1.159±0.210) and no parametrial invasion (1.718±0.710 vs 1.183±0.258), respectively (all P<0.05). The expression of NICD in cervical adenocarcinoma was higher than that of squamous cell cancer (1.463±0.395 vs 1.162±0.187, P<0.05). After SiHa and HeLa cells were incubated with DAPT, NICD expression was significantly lower than that in control (P<0.05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells was depended on its concentrations and times. Conclusions NICD may play a key role in the occurrence and progress of cervical cancer. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells may be due to decreased the formation of NICD.

Objective To discuss the surgical treatment for huge hepatoblastoma in children,and the technique of hepatectomy without blockade of the blood supply to the remained liver lobes.Methods We reviewed 12 cases of huge hepatoblastorna who had been operated from July 2001 to January 2009 in our hospital.The mean age of the children was 3.2 years(range,11 months to 12 years).The diameter of the tumor was from 10 to 23 cm.3～7 cycles of chemotherapy was routinely administrated before operation.When the tumor reduced to a certain size that radical resection could be performed safely,regular hepatectomy was conducted.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes was performed in every patient.Results The operations were successfully accomplished in all the 12 children.5 cases received right trihepatectomy (segment Ⅳ,Ⅴ～Ⅷ),4 cases received right hemihepatectomy(segment Ⅴ～Ⅷ),and the other 3 cases received Ⅳ,Ⅶ,Ⅷ segmentectomy,right Ⅴ,Ⅵ segmentectomy,and left hemihepatectomy respectively.The intraoperative hemodynamic parameters were stable,and there was no perioperative mortality.Postoperative chemotherapy wag routinely administrated.The follow-up period varied from 2 to 92 months.11 children survived and were disease free,among those 6 children have survived for more than 5 years.One child had brain and lung metastasis 5 months post operation,and died 7 months post operation. Conclusion Preoperative chemotherapy administrated to children with huge hepatoblastoma can reduce the tumor size and render tumor reseetable.Hepatoblastoma resection without blocking the blood supply to the remained liver lobes is a safe and feasible surgical technique.

Objective To determine the optimal procedure for preparation of the SPIO nanoparticles modified by dextran polylysine,and to evaluate the quality of this product.Methods The optimal preparation procedure of four factors (solution pH value,dextran concentration,reaction temperature and stirring rate) affecting SPIO hydrodynamic size were obtained through orthogonal experiments (3 levels).SPIO nanoparticles were synthesized under an optimum procedure.The shape and hydrodynamic size were detected by transmission electron microscope (TEM) and Malvern Zetasizer respectively.The content of Fe was detected by atomic absorption spectrophotometer,while chemical structures of SPIO nanoparticles were characterized and analyzed by infrared spectroscopy (IR) method.X-ray powder diffraction method was used to identify the ingredients,and the magnetic parameters were measured by vibrating sample magnetometer.Furthermore,experiments with ovarian cancer cells were performed to primarily validate the magnetic property of SPIO nanoparticles.Results The results of the orthogonal experiments showed that the optimum preparation procedure was as follows:dextran concentration of 10 mg/ml,pH 10,reaction temperature of 80 ℃ and stirring rate of 600 r/min.The TEM results showed the SPIO nanoparticles were in spherical shape,homogeneously distributed and uniform in size,and the mean diameter was 7.0 nm.The content of Fe was (12.36±0.08) g/L.The IR results clearly showed that the Fe3O4 was coated by dextran successfully.The Xray powder diffraction method showed that the sample contained Fe3O4 and the magnetism parameters indicated that the sample had superparamagnetism.The experiments with ovarian cancer cells demonstrated that SPIO nanoparticles could enter into the cells and then the cells had certain magnetic properties.Conclusions The SPIO nanoparticles synthesized under the optimal procedure are stable,small in size,with good dispersion and are feasible to enter into cells for rendering certain magnetic properties.This study has provided a good foundation and potential for further research.

Objective To evaluate the clinical application of perforator flap in extended radical vulvectomy of vulvar carcinoma.Methods Retrospectively,twelve cases of vulva carcinoma were treated by radical extensive excision,and the defects were repaired with perforator flap.Results All the flaps were survived and healed with first intention except one infection.The wound infection patient was treated with change of the dressing and antibiotics.The reconstructed vulvae were plump and elastic.It appeared like the normal vulvae and there was no contraction of the vagina.Conclusions Vulvar reconstruction with the perforator flap after the radical vulvectomy could make the patients recover easily,which produces almost normal appearance and function of the vulvae,reduces the time of wound healing,the patient could get the next therapy more quickly and the quality of life improving.It has wide clinical application value.

Objective To investigate the change of the neutrophil apoptosis and neutrophil respiratory burst in the patients and the effect of ulinastatin on the apoptosis of neutrophil and respiratory burst of neutrophil during cardiopulmonary bypass (CPB). Methods Sixty-two patients undergoing valve replacement with CPB were randomly divided into two groups: ulinastatin group (U group, 31 cases) and control group (C group, 31 cases). In U group patients received ulinastatin after induction of anesthesia. In C group patients received equal volume of normal saline, instead of ulinastatin. Arterial blood was obtained before operation (T1), 30 min after the start of CPB (T2), 30 min after the termination of CPB (T3). The apoptosis of neutrophil and respiratory burst of neutrophil were measured by flow cytometer. The level of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by kit. Results In C group, compared with T1 [(66.57±5.93)%], the rate of the apoptosis of neutrophil was significantly decreased at T2[(55.37±3.51)%] and T3 [(48.92±4.21)%] (P<0.05). And in U group, compared with T1 [(73.57±7.94)%], the rate of the apoptosis of neutrophil was significantly decreased at T2 [(68.34±4.92)% ] and T3 [(62.13±4.76)%] (P<0.05), And it reached to the minimum at T3. The rate of the neutrephil apeptosis was significantly lower in C group than that in U group (P<0.05). The respiratory burst of neutrophil increased significantly after the start of CPB and reached to the peak at T3[C group (1422.50±89.75) MCF,U group (1156.52±93.20) MCF]. The respiratory burst of neutrophil in U group was significandy lower than that in C group at T2 and T3 (P<0.05). The vitality of SOD decreased significantly after the start of operation in the two groups (P<0.05). The level of MDA increased significantly after the start of operation in the two groups, and reached to the peak at T3. The vitality of SOD in C group was significantly lower than that in U group at T3 (P<0.05). The level of MDA in C group was significantly higher than that in U group at T3 (P<0.05). Conclusions The rate of neutrophil apoptosis decreased and the respiratory burst of neutrophil increased during CPB. By improving the apoptosis of neutrophil and reducing the respiratory burst of neutrophil, ulinastatin can inhibit inflammatory reaction during CPB. Meanwhile, ulinastatin can improve the vitality of SOD and reduce the level of MDA. In conclusion, ulinastatin has a significant protective effect during CPB.