Objective To compare the efficacy of ultrasound ablation and laparoscopic lesion resection with uterine artery liga-tion in treating adenomyosis .Methods By using the prospective ,non-randomized controlled clinical study method ,the adenomyosis patients with clinical symptoms and requiring reserved uterus in our hospital from March 2011 to June 2012 were treated by the ul-trasound ablation technique(ultrasound ablation group ,40 cases) and the laparoscopic lesion resection with uterine artery ligation (operation group ,38 cases) .The menstrual blood volume ,dysmenorrhea and treatment satisfaction in postoperative1 ,3 ,6 months were compared between the two groups .Results The menstrual blood volumes in postoperative 1 ,3 ,6 months in the two groups were decreased significantly(P<0 .05) ,the dysmenorrhea symptoms were improved significantly (P< 0 .05) ,the two groups all were satisfied with the treatment effect .The differences between the two groups had no statistical significance (P>0 .05) .Conclu-sion The two methods of the ultrasound ablation technique and the laparoscopic lesion resection with uterine artery occlusion for treating adenomyosis could significantly reduce the menstrual blood volume ,improve dysmenorrheal and obtain higher satisfaction . Both can achieve the same therapeutic effect .

ObjectiveTo prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening hybridomas by indirect-ELISA.The two mAbs were named 3C3 and 5A5,and identified as IgG1 (κ) and IgG2(κ),respectively.They could react with NMP from M5-90.Conclusions Results of identification show that two mAbs against rBP26 can be produced.The two mAbs can recognize natural BP26 protein,giving the experimental materials for further research on identification of its epitopes.

Objective Our previous study demonstrated that toll-like receptor 2(TLR2) can distinguish extraneous antigen and prevent the immunological response. This study was designed to detect the expression of toll-like receptor 2 (TLR2) mRNA in cornea and investigate the effect of steroid on TLR2 expression in rats cornea following allograft penetrating keratoplasty. Methods The penetrating keratoplasty models were established in SPF rats with the 108 SD rats as receiptors and 36 SPF Wistar rats as donors, and other 6 SPF SD rats worked as normal controls. The receiptor rats were divided randomly into autograft group, allograft group and steriod group. The clarity and neovascularization of corneas of experiment rats were examined under the slit-lamp microscope and the rejection index was calculated based on Holland criteria. Corneal histopathological examination was carried out by hemotoxylin and eosin staining under the light microscope, and real time-PCR was employed for the detect of TLR2 mRNA in the corneas at the fifth, seventh and ninth day after operation. The experimental animals were obtained from the Animal Experimental Center of Southern Medical University and the procedure followed the Statement of Association for Research in Vision and Ophthalmology. Results The rejection occurred in 7 days after operation in allograft group, and only mild edema, opacity and neovascularization of corneas were found at different degrees in 9 days after operation in autograft group and steriod group. Severe corneal edema, a lots of inflammatory cells infiltration and new vessels in stroma were seen in allograft group, and mild inflammatory response was found in autograft group and steriod group. Normal comeal structure was exhibited in normal control group under the light microscope. The fold differences of TLR2 mRNA expression in cornea after amplification was significantly different among three groups and different time points (F_(group) = 39. 46, P = 0. 00; F_(time) =35. 38, P = 0. 00 ; F_(interaction) = 45. 66, P =0. 00), and the evident enhance of TLR2 mRNA expression was revealed in allograft group compared with autograft group (P ＜ 0. 05) and declined in steriod group (P ＜ 0. 05). Conclusion Steriod may restrain the acute allograft rejection by down-regulating the expression of TLR2 in corneas and its signals transaction. This result suggests that steriod offer a protection from rejection of cornea after penetrating keratoplasty.

ObjectiveTo retrospectively analyze the clinical characteristics and outcomes of endometrial carcinoma patients aged 45 years and younger MethodsFifty-two cases of endometrial carcinoma aged 45 years and younger were treated in Peking Union Medical College Hospital They were further divided into group A (35 years of age and younger) and group B (older than 35 years) Clinical data of these patients were reviewed and the two groups were compared ResultsPatients aged 45 years and younger accounted for 12 7% of all the endometrial carcinoma cases About 50% of the patients were nulliparous, infertile or had irregular menstruation and endometrial hyperplasia, 29% were obese, 23% had polycystic ovaries Eighty-three percent of the patients were stage [ Int ernational Federation of Gynecology and Obstetris (FIGO),1988] Group A had mo re polysystic ovaries and atypical endometrial hyperplasia than group B (53% v s 9%, 59% vs 26% respectively, P

Objective To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell(DC) transfected with interleukin(IL)-12 gene in vitro. Methods Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor ? were used to generate DC from cord blood in vitro. IL-12 fusion gene was cloned into a eukaryotic vector-pcDNA3.1(+). DC were transfected with IL-12-pcDNA3.1(+) using lipofectamine transfection method and electric transfection method respectively. Then polyethylene glycol mediated the fusion of transfected DC and ovarian carcinoma cells, and 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test was carried out to observe the immune response. Results DC were generated in vitro from cord blood and expressed high levels of costimulatory(50%) and MHC Ⅱ molecules(99%). RT-PCR showed that IL-12-pcDNA3.1(+) had been successfully transfected into DC. RT-PCR and immunofluorescence analysis showed that DC were fused with ovarian carcinoma cells successfully. Then the fusion cells of ovarian carcinoma cells to transfected DC, and the fusions of ovarian carcinoma cells to DC were cocultured with peripheral blood mononuclear cell respectively. MTT test showed that both fusions could induce cytolytic T cell activity and lysis of ovarian carcinoma cells,and the former effect was stronger. Conclusion The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.

OBJECTIVETo gain insight into the role of Toll-like receptor 2 (TLR2) in graft rejection following penetrating keratoplasty, and investigate the expression of TLR2 mRNA in the corneal graft.

METHODSPenetrating keratoplasty was performed in 3 groups of rats for orthotopic autologous corneal transplantation (group A), allograft corneal transplantation (group B), or allograft corneal transplantation with hormone treatment (C). The transparency and neovascularization of the cornea were observed using a slit-lamp microscope and scored according to the rejection index, with normal cornea serving as the control. The corneal tissues were sampled at 5, 7, and 9 days after the transplantation for histopathological examination and detection of TLR2 mRNA expression using RT-PCR.

RESULTSWith the passage of time, edema, opacities and neovascularization of the corneal graft occurred after the operation in all the groups. Seven days after the operation, the rejection index of group B, but not that of groups A and C, met the diagnostic criteria for graft rejection with also support by histopathological evidence. The expression of TLR2 mRNA was detected in normal corneas and augmented in the corneal grafts in the 3 transplantation groups. TLR2 mRNA expression in group B was significantly higher than that of group A, and the expression in group C decreased significantly in comparison with that in group B (P<0.05).

CONCLUSIONAs the recognition receptors of native immune system, TLR2 in the rejected corneal grafts may recognize the allograft antigen and play a role in acute graft rejection after penetrating keratoplasty.

Animals ; Cornea ; metabolism ; Female ; Graft Rejection ; immunology ; Keratoplasty, Penetrating ; Postoperative Complications ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism

BACKGROUND AND OBJECTIVECombined hypoxic cytotoxic drugs and chemoradiotherapy is an important mean of oncotherapy, and Tirapazamine (TPZ) is one of the most remarkable drugs. It has been shown that TPZ has a synergistic effect with radiotherapy on tumor cells, but whether TPZ would down-regulate the expression of the hypoxia-induced genes has not been reported. This study was to investigate the hypoxia-induced mRNA expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and osteopontin (OPN) in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells and the radiosensitization of TPZ, a hypoxia-specific drug, on HNE-1 and CNE-1 cells in vitro.

METHODSThe IC50 values of TPZ for HNE-1 and CNE-1 cells were measured using MTT assay, and the mRNA expressions of HIF-1alpha and OPN in HNE-1 and CNE-1 cells was determined using RT-PCR under aerobic and hypoxic conditions, respectively. The survival rates of HNE-1 and CNE-1 cells treated with or without TPZ at IC10 in the presence or absence of oxygen for 6 h were determined using colony formation assay following exposure to 1-6 Gy of 60Co radiation. The dose-survival curves were plotted and the values of D0, Dq and SER were calculated as a single-hit multitarget model.

RESULTSThe IC50 values of TPZ were 34.81 μmol/L and 35.02 μmol/L in HNE-1 and CNE-1 cells under aerobic condition, and 30.20 μmol/L and 28.48 μmol/L under hypoxic condition, respectively. The expressions of HIF-1alpha and OPN mRNA were reduced by TPZ in HNE-1 cells, but not in CNE-1 cells under hypoxic condition. For the HNE-1 cells, the respective values of D0 and Dq were 0.89 Gy and 0.28 Gy following normoxic irradiation versus 1.47 Gy and 0.44 Gy following hypoxic irradiation. For the CNE-1 cells, the respective values of D0 and Dq were 0.72 Gy and 0.68 Gy following normoxic irradiation versus 0.95 Gy and 0.56 Gy following hypoxic irradiation. The values of D0 and Dq for HNE-1 and CNE-1 cells treated with TPZ under hypoxic condition following irradiation were 0.66 Gy, 0.21 Gy and 0.85 Gy, 0.79 Gy, respectively.

CONCLUSIONTPZ can down-regulate hypoxia-induced expression of HIF-1alpha and OPN mRNA of HNE-1 cells and radiosensitize the HNE-1 cells but not CNE-1 cells, and act as a hypoxia modifier.

Antineoplastic Agents ; pharmacology ; Cell Hypoxia ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Cobalt Radioisotopes ; Down-Regulation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Inhibitory Concentration 50 ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Osteopontin ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Radiation Tolerance ; drug effects ; Radiation-Sensitizing Agents ; pharmacology ; Triazines ; pharmacology

OBJECTIVETo compare the relation between the preoperative functional magnetic resonance imaging (fMRI) with blood oxygen level dependent (BOLD) technique and intraoperative motor evoked potential (MEP) monitoring for cortical mapping of primary motor cortex in patients with tumors near the central area. And to determine whether non-invasive preoperative fMRI can provide results equivalent to those achieved with the invasive neurosurgical "gold standard".

METHODSA prospective study of 16 patients with various pathological tumors of the central area was conducted. Preoperative fMRI scans using the BOLD contrast technique in each patient were performed. An activation scan was achieved by using a motor task paradigm, which consisted of simple flexion-extension finger movements and finger-to-thumb touching in a repeating pattern. The anatomical structure was delineated by the T(1)-weighted three-dimensional fast spoiled gradient recalled sequence (3D/FSPGR) immediately afterward. The BOLD images were overlaid on the T(1)-weighted 3D/FSPGR images, and then co-registered to the neuronavigation system. The fMRI activations were documented by using a neuronavigation system in sequence, and compared to standardized intraoperative MEP monitoring, which included direct cortical electrical stimulation (DCES) or transcranial cortical electrical stimulation (TCES) or their combination. The compound muscle action potentials of forearm flexor and hand muscle responses were recorded during either TCES or DCES. Two techniques were compared to determine the accuracy for cortical mapping of primary motor areas with fMRI.

RESULTSOverall, the intraoperative MEP monitoring showed good correlation with fMRI activation in 92.3% of cases. The coincidence rate, however, was 100.0% between TCES and fMRI, and 66.7% between DCES and fMRI respectively. There was no statistically difference between two cortical mapping techniques, chi-square test of paired comparison of enumeration data, P < 0.01.

CONCLUSIONBOLD fMRI was a high sensitive and reliable technique to locate the position of the primary motor areas and their spatial relation with adjacent tumor, especially for the presurgical planning in patients with central area brain tumor.

Adolescent ; Adult ; Brain Neoplasms ; pathology ; physiopathology ; surgery ; Child ; Evoked Potentials, Motor ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; Motor Cortex ; pathology ; physiology ; Neuronavigation ; Oximetry ; Prospective Studies ; Transcranial Magnetic Stimulation ; methods

OBJECTIVETo evaluate the impact of lymphadenectomy on the relapse and survival of malignant ovarian germ cell tumor (OGCT).

METHODSThe clinical data of 102 OGCT cases treated in Peking Union Medical College Hospital from June 1980 to June 2003 were analyzed retrospectively. All the data about lymphadenectomy during primary and secondary surgery were collected, and other factors related to prognosis were also collected at the same time. Chi-squared test was applied in the univariate analysis related to relapse of disease. Cox model was applied in multivariate analysis related to relapse and survival of disease.

RESULTSPelvic and paraaortic lymph node metastasis was not significantly related to prognosis in primary and secondary treated patients. Lymphadenectomy showed no significant impact on disease relapse and survival. In the primary treatment, International Federation of Gynecology and Obstetrics (FIGO) staging, chemotherapy regimen, residual tumor and lymphadenectomy were the significant factors related to the relapse. After being stratified for the chemotherapy regimen, lymphadenectomy was not significantly related to the relapse in bleomycin +etoposide +cisplatin or cisplatin +vincristine +bleomycin regimen group, and lymphadenectomy could prevent relapse in no chemotherapy or other chemotherapy regimen group. In relapsed patients, only residual tumor was significantly related to survival time after relapse.

CONCLUSIONSPelvic lymph node metastasis is not the significant risk factor related to prognosis. Lymphadenectomy may have a beneficial effect on survival, although such effect is not significant. Although lymphadenectomy provides important information for prognosis, they provide little benefit to those patients already requiring chemotherapy based on the original operative findings. Lymphadenectomy should be performed to primary or relapsed patients by an expert surgical team.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Combined Modality Therapy ; Female ; Germinoma ; mortality ; pathology ; surgery ; therapy ; Humans ; Lymph Node Excision ; methods ; Lymphatic Metastasis ; Neoplasm Recurrence, Local ; mortality ; pathology ; surgery ; therapy ; Neoplasm Staging ; Ovarian Neoplasms ; mortality ; pathology ; surgery ; therapy ; Prognosis ; Retroperitoneal Space ; Retrospective Studies

OBJECTIVETo compare the clinical characteristics of three subcategories of laparoscopic hysterectomy: total laparoscopic hysterectomy (TLH) and two subcategories of laparoscopic-assisted vaginal hysterectomy (LAVH): LAVHs and LAVHb.

METHODSWe retrospectively analyzed the clinical data of 393 patients underwent laparoscopic hysterectomy, including TLH (n=178), LAVHa (n=177), and LAVHb (n=38), in our hospital from September 2002 to September 2005.

RESULTSMyoma and adenomyosis of uterus were the most common diseases in this study, accounting for 66.9%, 38.4%, and 52.6% in TLH group, LAVHa group, and LAVHb group, respectively. The mean surgery duration and blood loss were not significantly different between TLH group and LAVHa group (P > 0.05), but were significantly less in TLH group than in LAVHb group (P < 0.05). The bulk of uterus in TLH group was significantly bigger than in other two groups (P < 0.05). The incidence of major complications in the TLH group (9. 0%) was lower than in LAVHa group (14.1%) and in LAVHb group (18.4%), but without statistical significance. Conclusion Compared with LAVH, TLH is feasible to deal with bigger uterus with less blood loss and shorter surgery duration and without more frequent complications.

Endometriosis ; surgery ; Female ; Humans ; Hysterectomy ; adverse effects ; methods ; Hysterectomy, Vaginal ; adverse effects ; methods ; Laparoscopy ; adverse effects ; methods ; Myoma ; surgery ; Retrospective Studies ; Uterine Neoplasms ; surgery