Objective To construct pcDNA 3.1(+)/HPV 18 E 6 fusion gene and a single-codon mutation pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R fusion gene in eukaryotic expression vector and study the effects on proliferation and apoptosis of cervical carcinoma cell line Hela. Method HPV 18 E6 gene sequence and the single-point mutation HPV 18 E6 F49R or HPV 18 E6 F127R were amplified from total RNA of Hela cell line by reverse transcription- polymerase chain reaction ( RT- PCR), then the gene sequences were respectively inserted into pcDNA 3.1(+) vector to reconstruct recombinant plasmids which were transfected transiently into Hela cells. MTT and RT-PCR were used to test the expression levels of HPV 18 E6 and the growth of HeLa cells after transfected about 48 h. The proliferation and apoptosis of Hela cells were detected respectively by cell counting and AO/EB fluorescent vital staining. Results The pcDNA 3.1(+)/HPV 18 E6, pcDNA 3.1(+)/E6 F49R and pcDNA 3.1(+)/E6 F127R eukaryotic expression vectors were successfully constructed. The gene of HPV 18 E6 was discriminably detected in the HeLa cells which were transfected with the recombinant plasmids. After several days, the proliferation of Hela cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R plasmid were obviously inhibited and the apoptotic rates were significantly increased, then the proliferation of cells transfected with pcDNA(+)/HPV 18 E6 was rather increased slightly, and we could observe the phenomena of early apoptosis and the formation of thekaryopyknosis by fluorescent microscope in the cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R. Conclusion The eukaryotic expressing vectors encoding HPV 18 E6 F49R and HPV 18 E6 F127R provide fundamental basis for the further study on HPV 18 E6 mechanism as well as prevention and treatment of uterine cancer.

Objective To evaluate the effect of antler polypeptides from Cervus elaphus yarkandensis on MC3T3-E1 cells. Methods The concentration of antler polypeptides of Cervus elaphus yarkandensis was measured by BCA protein assay kit. MC3T3-E1 cells were cultured and analyzed by BCIP/NBT chromogenic alkaline phosphatase (ALP) staining kit. After being induced to form mineralized knot, the cells were stained by using Alizarin red staining. Three different concentrations (10, 1.0, 0.1 μg/mL) of antler polypeptides were analyzed by MTT method and micronutrients enzymes standard method to determine the effect of cell proliferation and ALP synthesis. Results The concentration of antler polypeptides was 0.07 mg/mL. The results of in vitro cell activity analysis showed that the positive rate of ALP was 90%and the mineralization knot was stained red. Compared with the control group, the different concentrations of antler polypeptides all showed the function of cell proliferation and the effect was dose-dependent after 3 d and 7 d. Compared with the control group, at the 3 d, three groups of antler polypeptides promoted synthesis and secretion of ALP (P<0.01) and the results showed a dose-dependent effect. Conclusion Antler polypeptides could obviously promote the proliferation of MC3T3-E1 cells and secretion of ALP, which indicated that antler polypeptides have certain effect on osteoporosis.

Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by a new PRGD/PDLLA/VPA composite conduit.Methods In this study from February,2012 to March,2014,PRGD/PDLLA/VPA nerve conduits were tested in the rat sciatic nerve transection model.At different periods after operation,its ability to promote nerve regeneration was evaluated by sciatic functional index(SFI),electrophysiology (CMAPs,NCVs) and histologic assessment.Forty rats were randomly divided into 4 groups (n =10),group A:PRGD/PDLLA/VPA,group B:PDLLA/VPA,group C:PRGD/VPA and group D:autograft.Results At 12 weeks after surgery,the SFI value of group A (-45 ± 3.19)and group D (-42 ± 3.01)were significantly higher than those of group B(-79 ± 3.06) and group C(-72 ± 2.07)(P ＜ 0.05);The CMAPs of group A (24.89 ± 5.01) and group D (25.39 ± 5.63) were significantly higher than those of group B(14.88 ± 3.11) and C(15.00 ± 5.54);the NCVs of group A (31.42 ± 2.43) and group D (31.50 ± 2.16) were significantly higher than those of group B (20.11 ± 2.39) and group C(21.00 ± 2.13)(P ＜ 0.05).At 12 weeks after surgery,the numbers of regenerated nerve in the tube of group A (258 ± 6.18) and D(259 ± 5.59) were significantly higher than those of group B (231 ± 5.00) and group C(230 ± 5.07)(P ＜ 0.05).There was no significant difference between groups A and D(P ＞ 0.05).Conclusion These results illustrated that this new PRGD/PDLLA/VPA conduit could significantly facilitate the regeneration of short nerve defect and recovery of motor nerve,which provides a new thought for treatment of peripheral nerve injury.

Objective To observe the improvement of postoperative pulmonary function and oxygen partial pressure during general anesthesia for open abdominal surgery with lung protective ventilation strategies and alveolar recruitment maneuvers. Methods Seventy patients who underwent selective open abdominal surgery were selected, and they were divided into standard ventilation group (tidal volume 8 ml/kg) and protective ventilation group (tidal volume 6 ml/kg, 5 cmH2O positive end-expiratory pressure, and alveolar recruitment maneuvers, 1 cmH2O=0.098 kPa) according to the random digits table method with 35 cases each. The airway pressure, blood pressure, pulse oxygen saturation (SpO2), end-tidal partial pressure of carbon dioxide (PETCO2) and adverse reactions were observed. The SpO2, partial pressure of O2 (PaO2) and pulmonary function before surgery and 1, 3, 5 d after surgery were measured. Results The respiratory rate, airway pressure and PETCO2 levels in protective ventilation group were significantly higher than those in standard ventilation group: (12.3 ± 2.1) times/min vs. (10.2 ± 1.0) times/min, (15.1 ± 2.8) cmH2O vs. (13.5 ± 2.3) cmH2O, (34.6 ± 2.1) mmHg (1 mmHg=0.133 kPa) vs. (32.1 ± 1.4) mmHg, and there were statistical differences (P<0.05). The SpO2 in 2 groups was maintained at 0.99. There was no statistical difference in the incidence of postoperative complications between 2 groups (P>0.05). The SpO2 and PaO2 levels at 1, 3 d after surgery in protective ventilation group were significantly higher than those in standard ventilation group:0.951 ± 0.018 vs. 0.936 ± 0.016 and 0.964 ± 0.018 vs. 0.949 ± 0.018, (74.8 ± 6.8) mmHg vs. (65.0 ± 6.2) mmHg and (79.6 ± 6.0) mmHg vs. (70.6 ± 5.3) mmHg, and there were statistical differences (P<0.05). The forced expiratory volume in 1 s (FEV1), percentage of the estimated value of FEV1, forced vital capacity (FVC) and percentage of the estimated value of FVC at 1, 3 and 5 d after surgery in protective ventilation group were significantly higher than those in standard ventilation group, the FEV1/FVC at 1 d after surgery was significantly higher than that in standard ventilation group, and there were statistical differences (P<0.05). Conclusions The lung protective ventilation strategy and alveolar recruitment maneuvers can improve the postoperative pulmonary function and oxygen partial pressure during general anesthesia for abdominal surgery. Low vital volume, appropriate positive end-expiratory pressure and recruitment maneuvers can protect the lung in general anesthesia patients.

Objective To improve the organization and management of the hospital,optimize the medical processes,improve medical quality and reduce medical costs.Methods A method of using multi-agent technology to hospital complex system modeling and simulation was introduced.The theory of complex system and its modeling method were simply introduced and the complex system characteristic and the medical processes of hospital were analyzed.Results The technology and fundation of multi-agent modeling were summarized.The steps of modeling and simulation for complex system of hospital were presented.The swarm simulation platform and the detail design of agent model in hospital were also presented.Conclusion It is proposed that the simulation research of hospital should use the methodology of multi-agent-based modeling and simulation,which is the research method of complex system.

Objective To study the inhibition of anti-PDGF on proliferation of cultured human retinal pigment epithelial cells(hRPE) in vitro.Methods hRPE were cultivated and were exposed to different concentrations of anti-PDGF(0,1?10-6,5?10-6,1?10-5,5?10-5 and 1?10-4 mg?L-1) respectively .Growth curves were measured with cell counting and the vitalities of cells were examined by percentage of vital cells and total cells.Using MTT staining colorimetric to measure the inhibitory rate.The changes of cell cycle of hRPE were collected and their growth were detected with FCM analysis and the morphological changes of cells were observed by light microscope and electron microscope.Results Anti-PDGF of 1?10-6mg?L-1 stimulated hRPE proliferation slightly.AntiPDGF at dosages ranging from 5?10-6mg?L-1 to 1?10-4mg?L-1 inhibited cell proliferation effectively in a dose-dependent and time-dependent manner(P

Animals ; Carbon Tetrachloride ; toxicity ; Dinoprostone ; blood ; Ginsenosides ; pharmacology ; therapeutic use ; Liver ; ultrastructure ; Liver Cirrhosis, Experimental ; drug therapy ; enzymology ; immunology ; Male ; Panax ; Phospholipases A ; biosynthesis ; genetics ; Phospholipases A2 ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; biosynthesis

OBJECTIVETo investigate the differences between clinicopathological features and prognosis of alpha-fetoprotein (AFP) negative (AFP < 20 ng/ml) and positive (AFP ≥ 20 ng/ml) hepatocellular carcinoma (HCC) patients.

METHODSClinicopathological data of 142 AFP-negative and 109 AFP-positive HCC patients who underwent RO radical hepatectomy in the Cancer Hospital of Chinese Academy of Medical Sciences between January 2006 and December 2011 were retrospectively reviewed and analyzed in this study.

RESULTSCompared with the AFP-negative patients, a higher female to male sex ratio, the later Barcelona Clinic Liver Cancer ( BCLC) stage, more liver capsule invasion and poorer Edmondson-Steiner grade were in the AFP-positive cases (P < 0.05 for all). Furthermore, the 1-, 3-, and 5- year overall survival rates were 94.4%, 82.4% and 61.0% in the AFP-negative group and 87.2%, 61.1% and 40.2%, respectively, in the AFP-positive group (P < 0.001). The multivariate analysis with Cox's proportional hazards model showed that AFP status, tumor size and Edmondson-Steiner grade are independent risk factors for survival of all the patients (P < 0.05) , and large tumor and Edmondson-Steiner grades III/IV are independent risk factors for worse survival in AFP-negative patients (P < 0.05). However, large tumor diameter was proved to be an independent risk factor leading to poor prognosis of AFP-positive cases (P < 0.05).

CONCLUSIONHigh levels of AFP indicate that the tumors are more malignant and with unfavorable prognosis.

Asian Continental Ancestry Group ; Carcinoma, Hepatocellular ; chemistry ; mortality ; pathology ; surgery ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; chemistry ; mortality ; pathology ; surgery ; Male ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Survival Rate ; alpha-Fetoproteins ; analysis

OBJECTIVETo study the effects of monocyte-macrophages (THP-1) in malignant transformation of human bronchial epithelial cells (BEAS-2B) cells induced by coal tar pitch (CTP) and the expression of TNF-α in the process of the cell malignant transformation.

METHODSBEAS-2B cells and THP-1 Cells were divided into four groups: coal tar pitch (CTP) group, benzo(a)pyrene [B(a)P] group, dimethyl sulfoxide (DMSO) group, BEAS-2B and THP-1 co-culture (co-culture group) group. Carcinogenesis model was established. The soft agar colony formation, chromosome aberrations and cell cycle tests were used to detect the cellular malignant transformation. The ELISA assay was utilized to measure the levels of TNF-α in the supernatant of CTP group and co-culture group.

RESULTSThe chromosome number abnormalities could be observed in early stage of the experiment (the 10th generation cells), which showed the increased ratio of aneuploid to polyploid, and the decreased number of diploid. The colony formation rate of co-culture group (the 20th generation cells) was 17.63‰ ± 0.97‰, which was significantly higher than that (13.94‰ ± 0.84‰) of CTP group and that (12.96‰ ± 1.62‰) of B(a)P group (P < 0.05). The proportion of S phase cells in the co-culture group was 44.49% ± 0.68%, which was significantly higher than that (38.19% ± 1.26%) of CTP group and that (36.41% ± 1.19%) of B(a)P group (P < 0.05). The TNF-α level in the co-culture group was significantly higher than that in CTP group (P = 0.001).

CONCLUSIONMonocyte-Macrophages can accelerate the malignant transformation of BEAS-2B cells induced by CTP and increase the expression level of TNF-α.

Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; Coal Tar ; toxicity ; Coculture Techniques ; Epithelial Cells ; cytology ; drug effects ; pathology ; Humans ; Macrophages ; cytology ; Monocytes ; cytology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo evaluate the endogenous vitamin D level and its correlation with bone mineral density (BMD) in children under 7 years old.

METHODSTotally 6 838 children who visited the Growth and Development Clinic due to "growth retardation, night terrors, hyperhidrosis, and dysphoria" were enrolled in the study. Serum 25-hydroxyvitamin D [25(OH)D] level was measured by chemiluminescence, whereas individual BMD was measured by quantitative ultrasound.

RESULTSAmong all subjects, serum 25(OH)D level was 34 ± 14 ng/mL, and the Z value of BMD was -0.49 ± 0.54. With increasing age, serum 25(OH)D level and BMD decreased gradually (P<0.01), and the detection rates for vitamin D deficiency and low BMD increased gradually (P<0.01). Compared with those with sufficient vitamin D, children with vitamin D deficiency had a significantly lower BMD (P<0.01) and a significantly higher detection rate for low BMD (P<0.01). 25-(OH)D level showed a positive linear correlation with BMD in children with vitamin D deficiency (P<0.01).

CONCLUSIONSPreschool and school-age children have severer vitamin D deficiency than infants. Vitamin D level may be correlated with BMD within a certain range.

Bone Density ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Vitamin D ; analogs & derivatives ; blood ; Vitamin D Deficiency ; epidemiology