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 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.

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.

Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.

Objective To examine characteristics of patients with blood urea nitrogen (BUN) levels higher and lower than the normal limit.Methods During January 2012 to January 2015,116 patients with upper gastrointestinal diseases were selected to study,according to the patient's blood urea nitrogen level,all the patients were divided into high BUN group and low BUN group,and there were 76 patients in the high BUN group,and 40 patients in low BUN group,compared the biochemical indices,gastrointestinal bleeding forrest grading and disease severity of the two groups,and univariate logistic regression analysis.Results The serum white blood cell count,blood urea nitrogen,creatinine and glycated hemoglobin levels in patients of high BUN group [(9 593±5 012)× 102/μl,368.1±162.3 mg/L,11.2±3.7 mg/L and 6.38±1.08％] were significantly higher than that of low BUN patients [(6 804 ± 2 087) × 102/μl,121.0 ± 39.3 mg/L,8.1 ± 3.2 mg/L and 5.51 ± 0.42 ％] (t =3.645～12.659,all P＜0.05),and the hemoglobin levels (87.3±35.1 g/L) of the patients in high BUN group was significantly lower than that of the low BUN patients (108.0 ± 31.2 g/L) (t=3.252,P=0.032).Logistic regression analysis showed the presence of hemoglobin and glycosylated hemoglobin levelst of wo groups of patients was significantly different (P＜0.05),and showed that showed the highest correlation with BUN.Gastrointestinal bleeding forrest hierarchical data of the two groups of patients showed no significant difference (P＞0.05).The proportion of patients with gastric ulcers of high BUN patients was significantly higher than that of the low BUN patients (x2 =39.655,P=0.000).Conclusion Patients with high expression of serum urea nitrogen had more severe upper gastrointestinal bleeding,and it is worthy of attention in the process of clinical diagnostic.

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 construct a lentiviral expression vector of peroxiredoxin2(PRDX2) RNA interference (RNAi) and to investigate the effect of siRNA of PRDX2 genes on the proliferation of human colonrectal cancer SW480 cell .Methods RNAi tar‐get sequences were designed and synthesized towards the PRDX2 gene sequences .The lentiviral vector pGC‐EGFP‐shPRDX2 was constructed and identified .The vector was transformed into SW480 cells ,and the transfection efficiency was evaluated by fluores‐cence microscopy .The expression of PRDX2 was detected with Quantitative real‐time PCR (qRT‐PCR) and Western blot in the transfected cells .Cell growth and colony forming ability were detected with MTT and plate cloning technique .Results PRDX2 gene lentiviral vector was successfully established and was proved by gene sequencing .The expression of PRDX2 in mRNA and pro‐tein was significantly reduced(P<0 .05) .The PRDX2 mRNA and protein expression in SW480 transfected with lentiviral were sig‐nificantly reduced (P< 0 .05) ,and the ability of growth and proliferation were significantly reduced(P< 0 .05) .Conclusion PRDX2 gene lentiviral vector could be a stable and reliable tool .The proliferation and growth of SW480 cells transfected by pGC‐EGFP‐shPRDX2 could be effectively suppressed ,which could facilitate further investigation of the roles of PRDX2 gene in the de‐velopment and progression of colorectal cancer .

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.