Objective To establish human hepatic carcinoma HepG2 cell line in which the expression of exogenous gene could be controlled by doxycycline. Methods The expression plasmid pTet-on was transfected into hepG2 cells by lipofectamine 2000.The stably transfected positive clones were screened with G 418 and subcultured. The expression level of luciferase induced by doxycycline at a concentration of 1 ?g/ml in stably transfected cells was detected one by one by transient transfection of plasmid pTRE-luc into the stably transfected cells. Results The basal expression activity of luciferase in cells of clone No 6 was 87, and the activity was 16764 after induction with doxycycline. Conclusion The HepG2/Tet-on cell line was a regulatory human hepatoma cell line, which is useful for the study of carcinogenesis and gene therapy of hepatic carcinoma.

ObjectiveTo investigate the effect of dexmedetomidine on the median effectivetarget effectsite concentration (EC50) of remifentanil required for preventing body movement in response to skin incision made under propofol sedation.MethodsForty ASA Ⅰ or Ⅱ patients aged 20-50 yr weighing 45-58 kg scheduled for elective breast tumor excision were randomly allocated into 2 groups ( n =20 each):group remifentanil (group R) and group remifentanil + demedetomidine ( group RD).Sedation was induced with propofol TCI at target plasma concentration of 3.0 mg/L in both groups.In group RD dexmedetomidine 1.0 μg/kg was infused iv over 10 min before start of propofol TCI,while in group R equal volume of normal saline was infused instead of dexmedetomidine.Remifentanil TCI was started with target effect-site concentration set at 3.0 and 2.5 μg/L in groups R and RD respective at 13 min after beginning of propofol TCI.Skin incision (3 cm in length) was made when the target concentrations of propofol and remifentanil TCI were reached.Body movement was assessed by a nurse not involved in this study.EC50 and 95％ confidence interval (CI) of remifentanil were determined by up-and-down technique.The target effect-site concentration was increased or decreased by 20％ depending on the response of the previous patient to skin-incision.ResultsThe EC50 of remifentanil for preventing body movement in response to skin incision performed under propofol sedation was 1.7 μg/L (95％ CI 1.5-1.9 μg/L) and 2.5 μg/L (95％ CI 2.2-2.7μg/L) in groups RD and R respectively.The EC50 of remifentanil was significantly lower in group RD than in group R.ConclusionDexmedetomidine 1.0 μg/kg can decrease EC50 of remifentanil for preventing body movement in response to skin incision made under propofol sedation.

BACKGROUND: Biological fixation refers to the treatment of coarse or porous prosthetic surface. It is favorable to "bone ingrowth" prosthesis to achieve long-term stability. Porous tantalum is the latest scientific product that appears suitable to prosthetic surface owing to its porous feature and has been attracting a great deal attention.OBJECTIVE: To investigate the therapeutic efficacy of a porous tantalum uncemented acetabular cup in acetabular revision arthroplasty.DESIGN, TIME AND SETTING: A retrospective case analysis was performed at the Department of Orthopedics, Changhai Hospital, Second Military Medical University of Chinese PLA between April and November 2006.PARTICIPANTS/MATERIALS: Sixteen patients (16 hips), 7 males and 9 females, aged 54-81 years old, who received treatment at the Department of Orthopedics, Changhai Hospital, Second Military Medical University of Chinese PLA were included in this study. Of these patients, 2 suffered from femoral neck fracture, 6 from aseptic femoral head necrosis, and 8 from osteoarthritis. Trabecular metal cup (Zimmer, Warsaw, Indiana) provided a titanium alloy bottom layer and porous tantalum-coated surface, which was realized by technical combination of bone trabecular tantalum and titanium alloy. The prosthesis contained an ultrahigh modulus polyethylene lining.METHODS: All patients underwent acetabular revision with modular porous tantalum uncemented acetabular cup. Prior to replacement, acetabular defects and femoral prosthesis were evaluated. A posterolateral approach of hip joint was made. Following acatabular prosthesis removal, bone cement was cleared with osteotome, simultaneously scar tissue, granulation tissue, and fibrous limiting membrane in the acetabular fossa were erased with curette to avoid further damage to sclerotin. Bacteriological examination and susceptibility test with respect to intraarticular tissue fluid confirmed aseptic loosening. Antibiotics were used in each patient to prevent infection. Antibiotics and low molecular heparin were used afterwards to prevent infection and thromboembolism, respectively. Patients could not load within 4 weeks but could walk with the aid of two canes within subsequent 6 weeks. One week after surgery, plain X-ray films were taken in the lateral plane.MAIN OUTCOME MEASURES: Prior to and 6, 12, and 24 months after surgery, Harris hip score and University of California Los Angeles (UCLA) activity score were assessed. The position of acetabular prosthesis was observed through an anteropostedor pelvic radiograph.RESULTS: The average follow-up time was 24 months (range 18-25 months). Harris hip score significantly increased from 32.6 points (range 21-54 points) prior to revision to 87.5 points (range 56-90 points) after revision (P < 0.05). UCLA activity score significantly increased from 3.8 points (2-5 points) prior to revision to 8.2 points (range 6-9 points) after revision (P<0.05). The anteropostedor pelvic radiographs taken in the last follow-up displayed no phenomena regarding non-stability including prosthesis aversion and subsidence and revealed that the in-growth of sclerotin surrounding the bone trabecular tantalum acetabular cup. Postoperative complications were found in none of 16 patients. By the last follow-up, there had been 14 cases who did not complain of pains while walking and 2 cases that had slight pain and slight limp while walking. Deep venous thrombosis or nerve injury did not appear afterwards. No cases needed another acetabular revision arthroplasty.CONCLUSION: For cases of failed primary fixation of artificial acetabular cup, application of porous tantalum uncemented acetabular cup can produces favorable results in acetabular revision if no bone defects exist.

Objective To investigate the effect of dexmedetomidine on bispectral index (BIS) value at loss of consciousness (LOC) caused by propofol given by target-controlled infusion (TCI).Methods One hundred and twenty ASA Ⅰ or Ⅱ patients,aged 20-50 yr,weighing 41-68 kg,scheduled for general surgery were randomly divided into 3 groups (n =40 each):propofol group (group P),dexmedetomidine 0.5 μg/kg + propofol group (group D1P) and dexmedetomidine 1.0 μg/kg + propofol group (group D2P).The patients in each group were randomly assigned into 5 subgroups ( n =8 each):groups P0-4 receiving TCI of propofol with the target effect-site concentration (Ce) set at0,1,2,3 and 4 mg/L respectively.Groups D1P0-4 received iv infusion of dexmedetomidine 0.5 μg/kg at a rate of0.05μg·kg-1 ·min-1 and TCI of propofol with the target Ce set at 0,1,2,3 and 4 mg/L respectively at 5 min after the end of dexmedetomidine infusion.Groups D2 P0-4 received iv infusion of dexmedetomidine 1.0 μg/kg at a rate of 0.1μg· kg- 1· min- 1 and TCI of propofol with the target Ce set at 0,1,2,3 and 4 mg/L respectively at 5 min after the end of dexmedetomidine infusion.Three minutes after TCI of propofol was started,OAA/S score and BIS value were recorded.The OAMS score ≤ 2 was defined as LOC.The EC50 and 95％ confidence interval of propofol for LOC and BIS50 and 95％ confidence interval at LOC were calculated by Probit analysis.Prediction probability (Pk) of BIS value at LOC was calculated using Smith method.Results Compared with group P,EC50 was significantly decreased,BIS50 was significantly increased ( P ＜ 0.05 or 0.01 ),and no significant change was found in Pk in groups D2 P and D1 P ( P ＞ 0.05).There were no significant differences in EC50,BIS50 and Pk between groups D2 P and D1P ( P ＞ 0.05).Conclusion BIS value can accurately predict the level of consciousness during anesthesia with dexmedetomidine and TCI of propofol,but BIS value is increased at LOC.

Objective To explore the related risk facts of pulm onary throm boem bolism (PTE ) and analyze the relation betw een PTE and the traum a or m edical behavior by investigating the cases of PTE . Methods Thirty-three cases w ere selected from Institute of Forensic Science (IFS) from 2000 to 2014. Results In 33 cases, 16 decedents w ere m ale, 17 decedents w ere fem ale;different degrees of dyspnea, chest tight-ness and syncope sym ptom s w ere the clinical m anifestation of the deceased; the throm bus w as mainly distributed in the left and right pulm onary arteries. The main source of em bolism w as the deep vein of low er lim b and the left probability w as higher. Traum a, lim ited position, operation and cardiovascular disease show ed high-risk factors of PTE; D-D im er test, hem olytic test and com puter tom ography pul-m onary angiography w ere the diagnostic tools for PTE . In som e cases, traum a and m edical m alpractice could be involved in the cause of death. Conclusion N on-typical clinical sym ptom s present in the m ost cases caused by PTE , and these cases alw ays show m any high-risk factors. The relation betw een PTE and injury or m edical behavior should be considered carefully in the forensic pathological practice.

Objective To investigate the mechanism by which HBx facilitates the proliferation of hepatoma cells. Methods The expression of plasmid pHA-HBx encoding full length of HBx was transfected into HepG2 cell lines and the transformed cells were identified by RT-PCR. The nude mouse model of transplanted human hepatoma HBx-transfected HepG2 cells was established. The expression of VEGF both in HepG2 cell-formed tumors and HBx-transfected cell-formed tumors in nude mice was examined immunohistochemically. Results RT-PCR showed that HBx gene was integrated into the genome DNA of HepG2 cells and amplified; The rate of tumor formation in nude mice both of HepG2 cells and HBxtransfected HepG2 cells was 100%; The growth speed of HBx-transfected tumors was much faster than that in control group; The average volume of HBx-transfected tumors and non-transfected tumors was 2. 86?0. 34 cm3 and 2.48?0.22 cm3 respectively after 8 weeks(t=1.905 ,P

Objective To investigate the effects of HBx on proliferation and apoptosis of human hepatocellular carcinoma cell line HepGz in vitro and in vivo. Methods The expression plasmids of pHA-HBx encoding full length of HBx gene was transfected into HepG2 cells with Lipofectamine 2000. The proliferation activity of transformed cells was measured by calculating the growth curve,population doubling time and by the 3H-TdR incorporation rate assay. The nude mouse model of HepG2 expressing HBX gene was established and the apoptosis rate of tumor cells was detected with TUNEL assay. Results RT-PCR indicated that the HBx gene was integrated into the genome DNA of HepG2 cells and transcripted. The growth curve and population doubling time manifested that the proliferation activity of transformed cells was much higher than that of control group cells. The apoptosis rate of tumor cells expressing HBx gene was much lower than that of control group cells. ConclusionHBx facilitates the proliferation activity of hepatoma cells in vitro and in vivo and inhibits the apoptosis of hepatoma cells induced by adriamycin in nude mice.

ObjectiveControl study on the clinical efficiency and costs of fast track surgery(FTS) and traditional method was carried out in colorectal cancer patients. Methodsone hundred colorectal cancer patients were randomLy selected, 50 cases treated with conventional therapy as control group and 50 cases treated with FTS programme. The postoperative initial venting time, the incidence of complications, the hospital stay and cost index were compared between the two groups. ResultsThe postoperative initial venting time was in advance and postoperative stay was obviously shortened in FTS group. Hospitalization expenditure in FTS group was lower than that in control group. Patients recovered quickly, the result was satisfactory. The complication was not significantly different between the two groups (P ＞ 0.05 ) . ConclusionsFTS treatment can accelerate postoperative rehabilitation and elevate clinical efficiency in colorectal cancer patients during operation period. FTS treatment is a safe and effective method.

BACKGROUND: Fibrous scaffolds prepared by electrospinning possess similar structure to extracellular matrix, and exhibit convenience in introducing bioactive ceramics into polymer matrices, which have distinct superiority in making bone tissue scaffolds materials.OBJECTIVE: To evaluate the bioactivity of gas-jet/electrospun nanosized hydroxyapatite (nHAP)/poly (3-hydroxybutyrate) (PHB) scaffolds for being used as bone tissue engineering scaffolds.DESIGN, TIME AND SETTING: The in vitro cytology experiment was performed at the State Key Laboratoty of Oral Diseases,Sichuan University between March 2008 and April 2009.MATERIALS: The gas-jet/electrospun PHB and 10% nHAP/PHB scaffolds were prepared.METHODS: Rats bone marrow-derived mescenchymal stem cells were incubated on the nHAP/PHB scaffolds were served as experimental group, incubated on the PHB scaffolds were as control group, and cells incubated on cell culture plate were consided to be blank control group.MAIN OUTCOME MEASURES: mRNA transcript expression of bone-related markers, including alkaline phosphatase, type I collagen, and osteocalcin, were quantified utilizing reverse transcription-polymerase chain reaction (RT-PCR) at day 14 after culture.RESULTS: After 14 days, the bone-related markers were expressed in three groups, and had higher transcript levels in the cells cultured on the nHAP/PHB scaffolds than those on the PHB scaffolds and cell culture plate.CONCLUSION: The gas-jet/electrospun nHAP/PHB scaffolds present good bioactivity in vitro and have the potential to be used in bone tissue engineering.

Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases (Erk1/2) were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ ( 68. 04 ± 2. 57 ) % vs ( 53.41 ±1.01)%] phase and increased the apoptotic cell fraction[ (19. 18 ±2.20)% vs (5.63 ±0.55)%, P <0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2(0.28 ±0.05 vs 0.81 ±0.07, P <0.05). Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.