The incidence of venous thromboembolism in patients with ovarian cancer is much higher than other gynecologic cancers. Approximate 20%of ovarian cancer patients have hypercoagulable status during different phases of their disease. Ovarian cancer itself can induce hypercoagulability, but meanwhile the over activated coagulation system may promote disease progression. Coagulation system disorder is one of the most important prognostic factors in ovarian cancer. Recently, hypercoagulability becomes a hot spot in the ovarian cancer research ifeld. This article reviews the mechanism of hypercoagulability, its clinical implication and correlated treatment in ovarian cancer patients.

BACKGROUND:In vitro isolation and purity technique of stem cells mostly depends on the identification of cell surface marker,such as monoclonal antibody adherent spreading method,flow cell sorting method and immunomagnetic beads sorting method,but the operation was complicated and the price was high.OBJECTIVE:To observe the biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells,which were isolated and cultured using the two-step method.DESIGN,TIME AND SETTING:The opening study was conducted at the Stem Cell Research Room of Xinjiang Medical University from March 2008 to March 2009.MATERIALS:Totally 10 amniotic fluid specimens were obtained from pregnant women who underwent prenatal diagnosis following 16-22 weeks of gestation or voluntarily induced abortion.With ultrasonic guidance,amniocentesis was performed to collect 20-40 mL amniotic fluid.METHODS:Human amniotic fluid-derived embryonic mesenchymal stem cells were isolated and cultured using the two-step method.Amniotic fluid was first centrifuged and incubated till spindle-shape cells were seen,with the presence of flbroblast-tike cell colonies.Supematant was moved to a new 25 cm~2 culture flask for further culture till spindle-shape fibroblast-like mesenchymal stem cell colonies.When 70% confluence,cells were digested,and incubated in α-MEM,supplemented with basic fibroblast growth factor,served as the first passage.MAIN OUTCOME MEASURES:Morphological changes in human amniotic fluid-derived embryonic mesenchymal stem cells of primary culture and subculture were measured.Karyotype,cycle,growth curve and colony formation ability of human amniotic fluid-derived embryonic mesenchymal stem cells were measured.Surface antigen and cytokine were examined using flow cytometry,immunofluorescence and RT-PCR.RESULTS:Human amniotic fluid-derived embryonic mesenchymal stem cells were successfully isolated and subcultured.During metaphase,primarily cultured amniotic fluid cells presented scattered spindle cells and flbroblast-like mesenchymal stem cell colonies every 7 days.Passaged cells completely adhered in 12 hours.Following 1 or 2 days of latent period,cells proliferated rapidly.About 90% confluence was observed following 6 or 7 days of culture.Cell arranged regularly,showing whirlpool-shape,radiated shape.Cells were spindle-shape,with unclear boundary.Chromosome karyotype of human amniotic fluid-derived embryonic mesenchymal stem cells was normal diploid.Growth curve showed "S" shape,but the two-step method reached a peak at (6.1±0.5) days,which was significantly rapid compared with the one-step method (7.2±0.6) days (P=0.035).Flow cytometry analyses showed that P3 cells at S phase took up (14±2.3)% using the two-step method,which was more than the one-step method (9.0±1.4)% (P=0.031).Low-density human amniotic fluid-derived embryonic mesenchymal stem cells were incubated for 7 days prior to cells formed scattered cell colonies.However,colony forming efficiency using the two-step method (15.0±2.3)% were significantly more than the one-step method (10.0±1.8)% (P=0.021).Flow cytometry results showed that human amniotic fluid-derived embryonic mesenchymal stem cells expressed CD44,CD29 and CD105,but were negatively for CD45,CD34,HLA-DR.Immunofluorescence suggested that Oct-4-positive cells were observed in amniotic fluid.However,the proportion of Oct-4-positive cells using two-step method (1.2±0.3)% was significantly greater than the one-step method (0.9±0.2)% (P=0.041).RT-PCR suggested that human amniotic fluid-derived embryonic mesenchymal stem cells obtained using the two methods expressed Oct-4.CONCLUSION:Human multipotent mesenchymal stem cells are present in human amniotic fluid.The two-step culture protocol could be a kind of high performance and simple protocol which may not interfere with the normal prenatal diagnosis procedure.

Fourteen compounds were isolated from Dalbergia odoriferae and purified by repeated column chromatography on silica and sephadex LH-20 gel and structurally identified by spectral analysis. These compounds were identified as 4, 9-dimethoxy-3-hydroxypterocarpan (1), medicarpin (2), 2', 4', 5-trihydroxy-7-methoxyisoflavone (3), 2', 3', 7-trihydroxy-4'-methoxyisoflavan (4), formononetin (5), 3, 8-dihydroxy-9-methoxypterocarpan (6), koparin (7), 3-hydroxy-9-methoxypterocarp-6a-ene (8), 2'-hydroxyformononetin (9), stevenin (10), 2', 7-dihydroxy-4', 5'-dimethoxyisoflavone (11), lyoniresinol (12), 2, 4-dihydroxy-5-methoxy-benzophenone (13) and neokhriol A (14). Compounds 1, 3, 4, 6, 8, 12 and 14 were isolated from this plant for the first time. Antibacterial activity assay showed that compound 4 had inhibitory effect on Ralstonia solanacearum.
Anisoles ; chemistry ; isolation & purification ; pharmacology ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Benzophenones ; chemistry ; isolation & purification ; pharmacology ; Chromatography ; methods ; Dalbergia ; chemistry ; Dextrans ; Gels ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; Microbial Sensitivity Tests ; Naphthalenes ; chemistry ; isolation & purification ; pharmacology ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Pterocarpans ; chemistry ; isolation & purification ; pharmacology ; Ralstonia solanacearum ; drug effects ; growth & development ; Silica Gel

Objective To investigate the killing effect of polymethylmethacrylate (PMMA) on spinal metastasis of transplanted VX2 carcinoma in experimental rabbit models. Methods Spinal metastasis of transplanted VX2 carcinoma model was successfully established in 18 rabbits. The experimental rabbits were randomly and equally divided into three groups with 6 rabbits in each group. Under CT guidance , PMMA or saline was injected into the center of VX2 tumor; in group A 0.3 ml of PMMA was used, in group B 0.1 ml of PMMA was used and in group C (control group) 0.3 ml saline was used. Twenty-four hours after the injection, the animals were sacrificed. Four tissue samples were obtained from the sites at 1 mm , 5 mm, 10 mm and 15 mm away from the PMMA mass in each rabbit of group A and group B , while four tissue samples were collected from different four sites from the tumor ’s center to border in each rabbit of group C. TdT-mediated dUTP nick-end labeling (TUNEL) method was used to determine the tumor cell apoptosis rate. Results After successful establishment of rabbit model, injection of PMMA was performed in sixteen among the eighteen rabbits. Technical success rates were 83.3% in both group A and B, and the success rate was 100% in group C. The difference in technical success rate was not significant. The mean tumor cell apoptosis rates of spinal VX2 carcinoma at 1 mm, 5 mm and 10 mm away from the PMMA mass in group A were (65.75±18.81)%, (50.00±14.24)% and(14.95±8.98)% respectively. The mean apoptosis rate in the control group was (9.79 ±5.24)%; the differences between the group A and the control group were statistically significant (P<0.05). The mean tumor cell apoptosis rate of spinal VX2 carcinoma at 15 mm away from the PMMA mass in group A was (10.30 ±8.13)%, which was not significantly different with that of the control group. The mean tumor cell apoptosis rates of spinal VX2 carcinoma at 1 mm and 5 mm away from the PMMA mass in group B were (49.20±15.57)% and(17.75±9.28)% respectively, which was significantly different with that of the control group(P<0.05); the mean tumor cell apoptosis rates at 10 mm and 15 mm away from the PMMA mass in group B were not significantly different with those of the control group. Statistically significant differences in the mean tumor cell apoptosis rates determined at 1 mm, 5 mm and 10 mm away from the PMMA mass existed between group A and group B(P<0.001). Conclusion PMMA can promote the apoptosis of tumor cells, properly increasing the injected amount of PMMA can enlarge the extent of tumor cell apoptosis.

Objective To evaluate the effect of temporal resolution of breast dynamic contrast-enhancing (DCE)-MRI on pharmacokinetic parameters and diagnostic performance in benign and malignant breast lesions.Methods Retrospective review was performed on 26 benign and 29 malignant breast lesions which were proven pathologically by surgery or biopsy.Dynamic contrast enhanced breast MRI using a new volume-interpolated-breath-hold examination sequence combining parallel acquisition, Dixon fat separation and time-resolved imaging with interleaved stochastic trajectories (CDT-VIBE) was performed in all patients.The original time resolution of this sequence was 12 s and based on which, dynamic sequences with different temporal resolutions of 24 s, 36 s, 48 s and 60 s were simulated by a sample-removing method and were used for the calculation of pharmacokinetic parameters, including volume transfer constant (Ktrans), constant flux rate (Kep), volume of extravascular extracellular space (Ve) and area under the curve at initial 60 s (iAUC), to observe their changes with different temporal resolution.The repeated measurement of analysis of variance was used to explore significant changes in pharmacokinetics parameters with different temporal resolution.To evaluate the diagnostic efficiency of parameters with different temporal resolution, ROC analysis was performed in accordance with pathological findings.Results With decrease of temporal resolution from 12 to 60 s, there was a significant increase in Ktrans and Kep of benign lesions [Ktrans: (0.147±0.084)/min to (0.170 ± 0.085)/min, Kep: (0.321± 0.176)/min to (0.433± 0.175)/min] and steady decrease in Ktrans and Kep of malignant lesions [Ktrans: (0.373±0.210)/min to (0.259± 0.122)/min, Kep: (0.929 ±0.402)/min to (0.581 ± 0.143)/min];the changes of Ve were less significant and irregular;the values of iAUC decreased both in benign [(9.192± 4.660) to (7.388± 3.065)] and malignant [(20.221±9.876) to (12.850±5.194)] lesions.The changes in all parameters with different time resolution were statistically significant in benign and malignant lesions (all P＜0.05).By pair-wise comparison between different time resolution, the changes of K among 12 s, 24 s and 36 s (all P＜0.05), the changes of Kep between 12 s and others (all P＜0.01), the changes of Ve between 24 s and others (all P＜0.01), and the changes among all pairs of iAUC were significant (all P＜0.05) except for 12 s and 24 s (P=1.000).The best AUC value of Ktrans and Kep between benign and malignant lesions were achieved with 12 s dynamic sequences (0.887, 0.939), and the best AUC value of iAUC was with 24 s sequences (0.877).The AUC values of Ve were between 0.511 to 0.601, which was lower than that of other three parameters.Conclusion A 24 s or higher temporal resolution of DCE-MRI should be able to provide consistent differentiation of pharmacokinetic parameters in benign and malignant breast lesions.

Objective To describe the general characteristics of patients with acute chest pain in order to analyze factors associated with patients’utilization of emergency medical services (EMS).Methods A total of 747 eligible patients with acute chest pain admitted to emergency department of Qilu Hospital were consecutively enrolled from October 2014 to April 2015.Clinical data including demographic features, mode of arrival,past medical history,risk factors,symptoms and signs were collected prospectively by using standardized case report form.Univariate and multivariate analyses were carried out to investigate the association between the decision to use EMS and related factors including demographic features,past medical history,risk factors,symptoms and signs.Results Of the total 747 eligible patients,414 (55.4%)were male ,and the mean age was (57.2 ± 15.8)year;333 (44.6%)were female,and the mean age was (61.7 ±14.9)year.Of them,171 (22.9%)patients used EMS,and 143 chest pain patients with more than 75 years old were more inclined to use EMS (P <0.01),whereas 152 patients in 65 -75 years age group accounted for the lowest proportion of using EMS.Men were more inclined to use EMS than women (P <0.05),and 483 patients with typical chest pain used more EMS than patients with atypical chest pain (P <0.05);Of them,356 patients with a history of hypertension and 54 patients with a history of cerebral infarction were more inclined to use EMS (P <0.05 and P <0.01,respectively).Multivariate logistic regression analysis showed that male,older than 75 years,history of cerebral infarction were independent factors associated with EMS use (P <0.05).Conclusions This study indicated that only less than one-third of emergency department visits with acute chest pain decide to use EMS when symptoms occurred. Factors including male,older than 75 years,and a history of cerebral infarction were associated with more use of EMS.In order to promote patient asking for EMS timely,more work should be done.

Objective To compare the local control and survival rates of hyperfractionated radiotherapy plus concurrent chemotherapy with hyperfractionated radiotherapy alone in the treatment of stage Ⅲ-Ⅳ nasopharyngeal carcinoma (NPC).Methods Between December 1992 and December 1995, 150 NPC patients were randomized into hyperfractionated radiotherapy plus concurrent chemotherapy (R+C) and hyperfractionated radiotherapy alone (R alone) groups. Radiotherapy were similar in the two groups: 1.2 Gy/f, twice a day. Chemotherapy was given to R+C patients before and during the course of radiotherapy. Results The overall 5-year survival (OS), disease-free survival and distant metastasis-free survival rates were 57.3%, 55.9% and 55.9% . The 5-year survival rates of the R+C and R alone groups were 64.0% and 50.7%, with the difference statistically significant (P=0.037). One patient in the R+C group and 5 patients in the R alone group developed nasopharyngeal recurrence and the corresponding 5-year local control rates were 98.7% and 93.4%. The acute mucosal reaction in the R+C patients was severer than that of the R alone, but well tolerated and did not develop any severe complications. Conclusions Hyperfractionated radiotherapy plus concurrent chemotherapy can improve the local control and survival in patients with stage Ⅲ-Ⅳ nasopharyngeal carcinoma with well tolerated mucosal reactions. Chemotherapy gives greater benefit on the survival of stage Ⅳ patients.

Objective To isolate enterovirus 71 from a death children,and analyze whether the neurovirulence was related to the variation of nucleotide and amino acid. Methods Enterovirus 71 was isolated from throat swabs which were colleted from Shandong Linyi People's Hospital. The full length genome was sequenced by amplification with RT-PCR and sequencing of 9 overlapped gene fragments covering full length of the genomes. The nucleotide and amino acid sequenced was aligned by BLAST, Bioedit and MEGA 4. Results A strain of enterovirus 71 was isolated and named as SDLY107. The full length was 7405 bp. The results of homology analysis of overall nucleotide sequence showed that strain Fuyang. Anhui. P. R. C/17.08/2 had highest homology (98.6%)with strain SDLY107, and the homology was 80.0% between strain SDLY107 with prototype strain BrCr/70,and 86. 5% between strain SDLY107 with nerve strain MS/87. Phylogenetic analysis showed that the phylogeny was close between SDLY107 with some isolated strains from Chinese Mainland, such as Beijing, Henan, Guangxi, Sbenzhen, Lanzhou, Fuyang, Chongqing and Zhejiang strains, which was clustered for C4 subtype. The results of amino acid sequence analysis showed that there were 2 mutations, E947D and K1873R, for strain SDLY107. Conclusion SDLY107 belonged to C4 subtype, amino acid mutations E947D and K1873R of which may be relevant to the pathogenicity of EV71.

To construct the lentiviral vector containing Peroxiredoxin 2(Prdx2) gene and the colorectal cancer cell line stably transduced with Prdx 2-containing vector , so as to provide a useful tool for studying the role of Prdx 2 in colorectal cancer.Methods: Prdx2 was amplified by PCR and inserted into lentiviral expression vector Ubi-MCS-EGFP-IRES-Puromycin (GV218) to generate Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector.The inserted Prdx2 gene was verified by double enzyme digestion and DNA sequencing.Subsequently ,lentiviruses were produced and transduced into SW 480 cells.EGFP expression was examined under fluorescence microscopy ,the expression of Prdx2 was detected with qRT-PCR and Western blot.Cell growth and colony forming ability were detected with MTT and plate cloning technique.Results: The lentiviral Prdx2 expression vector was successful construc-ted.Overexpression of Prdx2 was verified in SW480 cells with LV-Prdx2 vector.Prdx2 promoted SW480 cell growth and colony forming ability(P<0.05).Conclusion:Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector is successfully constructed,and the SW480/LV-Prdx2 cell line with stable transduction of Prdx2 containing vector is established.Overexpression Prdx2 can significantly promote the proliferation of colorectal cancer SW 480 cells.

Objective Gambogic acid ( GA) can suppress the growth of multiple tumor cells , including gastric carcinoma , hepatoma , hematologic neoplasms and breast carcinoma , but there have been few reports about its effect on urologic neoplasms .This study was to investigate the possible mechanisms of GA inducing bladder cancer cell apoptosis and cell cycle arrest . Methods We cultured human bladder cancer BIU8-7 cell lines in vitor and treated the cells in the logarithmic growth phase with isotonic saline solu-tion (negative control)or GA at the concentrations of 1.0, 2.0, and 3.0μmol/L, respectively.We determined the expression of the Caspase-3 protein in the tumor tissue using the immunohistochemical S-P method and detected GA-induced apoptosis of the bladder cancer cells and cell cycle changes by flow cytometry . Results The expressions of the Caspase-3 protein were 4.28 ±1.86, 5.03 ± 0.78, and 6.47 ±1.31 in the 1.0, 2.0, and 3.0μmol/L GA groups, respectively, significantly higher than 2.13 ±1.27 in the nega-tive control (P<0.05).Flow cytometry showed a gradual decrease of the cells in the G 0/G1 phase and a gradual increase in the G2/M phase , but no obvious change in the S phase . Conclusion Gambogic acid can promote the apoptosis , arrest the cell cycle , and in-hibit the proliferation of bladder cancer cells by increasing the expression of the Caspase -3 protein.