Cell Culture Techniques ; Endothelial Cells ; cytology ; Humans ; Lymphangiogenesis ; physiology ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Neoplasms ; blood supply ; metabolism ; physiopathology ; Vascular Endothelial Growth Factor C ; metabolism ; physiology ; Vascular Endothelial Growth Factor Receptor-3 ; metabolism

Objective To establish a malignant transformed human fetal osteoblastic human cells from the immortalized cell line hFOB1.19 in order to explore the molecular mechanism in tumorigenesis of osteosarcoma. Methods hFOB1.19 cells were treated sequentially by an initiated factor, N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) and a promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). The features of malignancy of transformed cells were identified by cell morphology, DNA content analysis, colony forming frequency on soft agar and tumorigenetic test in nude mice. Results Continuous passaging after the treatments resulted in the formation of a few paramorph foci and exhibited in an extensively random orientation. The poly-ploid of DNA was 81.08% in experiment group, much higher than that in control group (55.03%). Compared with that of negative control cell, colony formation efficiency of transformed cells in semisolid agar showed a a significant increase. The transformed cells formed tumors subcutaneously in the nude mice, which were verified to be poorly differentiated osteosarcoma histopathological examination. Conclusion Mocking the process of malignant transformation of human cells, we establish malignant transformed immortalized human fetal osteoblastic human cells (hFOB1.19) model.

Background Congenital cataract is an important cause of blindness and amblyopia in children,and about 50％ of congenital cataract is hereditary.Objective The aim of this study was to determine the diseasecausing gene of one Hui congenital cataract pedigree by using exon combined target region capture sequencing chip of eye diseases.Methods This study was approved by Ethic Committee of Ningxia People's Hospital and followed Declaration of Helsinki.One Hui congenital cataract pedigree was recruited in Ningxia Eye Hospital in 2011.All the disease history of the members in this family were collected and recorded,and the eye examinations were performed.The peripheral blood specimens were collected from family members and 300 healthy individuals for the extraction of DNA.Exon combined target region capture sequencing chip of eye diseases was used to screen the candidate diseasecausing mutations,then PCR and direct sequencing were used to confirm the disease-causing mutations.Results This H ui family included 61 members of 6 generations,and 18 patients were diagnosed in serial 5 passages,conforming to autosomal dominant inheritance pattern.Among 18 cataract patients,7 individuals were associated with nystagmus and strabismus,and 4 patients had high myopia.Eight candidate pathogenetic mutations were detected by exon combined target region capture sequencing chip of eye diseases and bioinformatics method,with 5 mutations in noncoding regions and 3 in coding regions.The mutation P24T of CYRGD gene was confirmed as pathogenic mutation of this pedigree by using PCR and direct sequencing methods.These mutations co-segregated with affected members of the family,and the mutations were not found in the unaffected family members and 300 unrelated controls.Conclusions P24T of CYRGD gene mutation is confirmed as pathogenic mutation of this pedigree.Exon combined target region capture sequencing chip provides a new approach to detect disease-causing mutations of congenital cataract with diversity clinical phenotypes.

Objective To discuss the risk factors of C5 never palsy incidence after cervical open-door laminoplasty. Methods This study involved 254 patients with cervical spondylotic myelopathy, who underwent expansive open-door lam?inoplasty in our hospital between January 2009 and December 2012. Patients were divided into two groups according to the incidence of C5 palsy after operation. Group A included 20 patients with C5 palsy and group B included 234 cases without C5 palsy. The risk factors for postoperative C5 palsy were compared between two groups. The factors may affect the inci?dence of C5 palsy were analyze. Results All cases received follow-up with an average follow-up time (24.9±2.6) months. Values of cervical curvature index (CCI), lamina open angles and spinal cord shift were significantly greater in group A than those of group B (P<0.05). There were no significant differences in preoperative intramedullary high signal, ossification of posterior longitudinal ligament and intervertebral foramen stenosis between two groups (P>0.05). Results of Logistic regres?sion analysis of multiple factors showed that lamina open angles, the pathogenic factors of C5 palsy included the spinal cord shift and postoperative CCI [OR(95%CI):2.035(1.684-2.359), 3.542 (2.465-9.254) and 9.126(4.425-17.273). Conclusion The angle of open door too large, CCI and spinal cord shift are risk factors for postoperative C5 palsy.

Objective To investigate the changes in cerebral oxygen metabolism during liver transplantation in patients with liver cirrhoses.Methods Sixteen ASAⅢorⅣpatients with liver cirrhoses(14 male,2 female)aged 25-67 yrs,weighing 45-80 kg undergoing liver transplantation were studied.Radial artery was cannulated for direct BP monitoring and blood sampling.Swan-Ganz catheter was placed in pulmonary artery (PA)via right internal jugular vein(IJV)for cardiac output(CO)monitoring and sampling mixed venous blood. Left IJV was cannulated and the catheter was advanced cephalad until jugular bulb for blood sampling.Anesthesia was induced with midazolam,fentanyl,propefol and vecuronium and maintained with isoflurane inhalation and intermittentⅣboluses of fentanyl and vecuronium.The patients were mechanically ventilated after tracheal intubation.PaCO_2 was maintained between 30-45 mm Hg.Blood samples were taken from radial artery,pulmonary artery and jugular bulb simultaneously for blood gas analysis before operation(T_0,baseline),10 min before anhepatic phase(T_1)20 min after onset of anhepatic phase(T_2),30 min after graft reperfusion(T_3)and at the end of operation(T_4).Oxygen delivery(DO_2),oxygen consumption(VO_2),oxygen content of jugular bulb blood (CjvO_2),cerebral arterial-venous oxygen content differences(Ca-jvO_2)cerebral oxygen extraction ratio(CERO_2) and CBF/CMRO_2 were calculated.Results The mean duration of operation was(364?51)min and the mean intraoperative blood loss was(1340?430)ml.CO was significantly increased before anhepatic phase(T_1), during neohepatic phase(T_3)and at the end of operation(T_4)but decreased during anhepatc phase(T_2)as compared with the baseline value at T_0.Hb,CaO_2,Ca-jvO_2 and CERO_2 were all decreased while SjvO_2 and CBF/ CMRO_2 were increased during operation;DO_2,VO_2 and CjvO_2 were decreased during anhepatic phase;DO_2 was increased during other phases;VO_2 was increased at the end of operation as compared with the baseline(T_0)(P＜0.05 or 0.01).Conclusion There is no cerebral oxygen deficiency during liver transplantation in patients with liver cirrhoses.

Objective To study the effect of trachea intubation and mechanical ventilation on the prognosis and discharge rate of patients with successful cardiac-pulmonary resuscitation.Method The clinical data of 389 patients,who were admitted from January 2005 to February 2007,were retrospectively analyzed.The relation between trachea intubation time and discharge rate was studied.According to the time from cardiac arrest to finishing trachea intubation,patients were divided into group A (within 3 minutes,n=209) and group B (over 3 minutes,n=143);according to the time from reaching emergency medicine department to finishing trachea intubation,the rest patients were divided into group C (within 5 minutes,n=9) and group D (over 5 minutes, n=38) minutes.The discharge rate was calculated between groups.The software of SPSS 11.0 was used for statistical analysis.Results The successful rate was 9.75% (389/3988),and 59 patients were discharged, with discharge rate 1.48% (59/3988).The discharge rate of group A was 19.62% (41/209),and was significantly higher than that of group B 6.99 % (10/143) (P

Objective To study the expression of anticentromere antibody (ACA)on multiple diseases to help the clinical diag-nosis and treatment.Methods The clinical and laboratory data of 129 cases with positive ACA were retrospectively ana-lyzed.Results ①116 females and 13 males were found among 129 cases with positive ACA,and serology titer of ACA in fe-male patients was higher than male and more compound antibody were shown in female patients,which was significantly dif-ferent compared with males (χ2=6.28,P=0.01;χ2=6.85,P=0.003).②Among 118 cases which had detailed clinical in-formation,there were 65 patients suffered from autoimmune diseases (55.08%)and 53 cases suffered from non-autoimmune diseases (44.92%).Autoimmune disease cases showed more compound and high serology titer,there were significant differ-ence between the two groups (χ2=21.97,P<0.001;χ2=11.44,P=0.001).Conclusion Though ACA was rarely found,it was shown in autoimmune and non-autoimmune diseases and can be not too severe or fetal and even lead to multi-organ fail-ure.So it should be taken seriously.

Objective: To denvolop HPLC an method for the determination of the content of three saponins in Zhixiao suppositories. Methods: Analysis carriedout was performed on a Kromasil C18 column (4.6 mm×250 mm., 5μm)at 23℃. The mobile phases were methanol and 0.2% phosphoricacid solution in a gradient elution. The flow rate was set at 1.0 mL·min-1 and the ultraviolet detector was operated at 203 nm. Results: The linear ranges of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 were 0.368～2.76μg,0.676～5.07 μg and 0.812～6.09μg,respectively. The average recoveries of the three saponins were 98.53%, 99.20% and 99.35%, respectively. The RSD (n=5) were 2.11%, 1.88% and 2.38%, respectively. Conclusion: The separation is a good and the results are accurate and reproducible. It can be used for the quality control of Zhixiao suppositories.

OBJECTIVETo investigate the changes of CD4 + CD25 + Foxp3 + regulatory T cells in the peripheral blood mononuclear cells (PBMC) and their association with insulin resistance in different stages of prostate cancer (PCa).

METHODSUsing flow cytometry, we counted the CD4+ CD25 + Foxp3 + regulatory T cells in the PBMCs of 62 PCa patients (5 cases of TNM stage I, 16 cases of stage II, 21 cases of stage III, and 20 cases of stage IV) and 42 normal healthy controls, and calculated their proportion in the CD4+ T-lymphocytes. We determined the levels of fast blood glucose (FBG) and fast insulin (FINS) for the insulin resistance index (HOMA-IR), obtained the serum IGF-1 level by ELISA, and analyzed the relationship of the count and proportion of CD4+ CD25+ Foxp3+ regulatory T cells with insulin resistance by comparison between the PCa patients and normal healthy controls.

RESULTSCompared with the control group, the PCa patients showed significantly increased HOMA-IR (3.68 ± 1.42 vs 6.68 ± 1.66), decreased level of serum IGF-1 ([164.56 ± 30.58] vs [96.39 ± 21.21] ng/ml), and elevated count ([1.99 ± 0.78 ] x 10(7) vs [3.55 ± 0.29] x 10(7)) and proportion ([5.33 ± 0.65] vs [13.88 ± 0.96]%) of CD4 + CD25 + Foxp3 regulatory T cells in the PBMCs. The TNM stage was correlated positively with the count and percentage of CD4 + CD25+ Foxp3 + regulatory T cells and HOMA-IR, but negatively with the level of serum IGF-1. Meanwhile, the count and percentage of CD4 + CD25 + Foxp3 + regulatory T cells were found to have a positive correlation with HOMA-IR (r = 0.722 and 0.689, P < 0.01) but a negative correlation with the level of serum IGF-1 (r = -0.747 and -0.896, P < 0.01).

CONCLUSIONThe count and proportion of CD4+ CD25 + Foxp3 + regulatory T cells in the peripheral blood and insulin resistance increase with the elevated stage of PCa. CD4 + CD25 + Foxp3 + regulatory T cells may be involved in the occurrence and progression of PCa by regulating insulin resistance.

Aged ; CD8-Positive T-Lymphocytes ; Case-Control Studies ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Hyperinsulinism ; Insulin ; blood ; Insulin Resistance ; Insulin-Like Growth Factor I ; metabolism ; Leukocytes, Mononuclear ; Lymphocyte Count ; Male ; Prostatic Neoplasms ; blood ; immunology ; pathology ; T-Lymphocytes, Regulatory

Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Carcinoma, Merkel Cell ; metabolism ; pathology ; Carcinoma, Small Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymphoma ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism