Objective To determine the 95 ％ effective target plasma concentration (EC95) of remifentanil for tracheal tube tolerance during the recovery period from anesthesia following cervical spine surgery.Methods Thirty ASA Ⅰ or Ⅱ patients,aged 18-60 yr,weighing 50-80 kg,scheduled for elective cervical spine surgery under total intravenous anesthesia,were enrolled in this study.Anesthesia was induced with iv injection of propofol,sufentanil and rocuronium.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propofol and target-controlled infusion of remifentanil.The target plasma concentration (Cp) of remifentanil was set at 4-6 μg/L.BIS value was maintained at 40-60.Infusion of propofol was stopped at the end of surgery.Participants were allocated to a dose of remifentanil by 3-patient cohorts.Six Cps were selected from 1.0-3.5 μg/L before beginning and they were 1.0,1.5,2.0,2.5,3.0,3.5 μg/L.The Cp of remifentanil was 3.0 μg/L in the first cohort.After completion of the trial in each cohort,the posterior probability of each concentration was calculated according to the condition of sedation/analgesia and anterior probability of each concentration.The concentration with the posterior probability closest to 95 ％ was chosen as Cp in the next cohort.The concentration-probability curve was made according to the posterior probability of each concentration,and then EC95 and 95 ％ confidence interval of remifentanil were calculated.Results The EC95 and 95 ％ confidence interval of remifentanil were 2.77μg/L (2.65-2.83 μg/L) for tracheal tube tolerance during the recovery period from anesthesia following cervical spine surgery.Conclusion The EC95 of remifentanil for tracheal tube tolerance during the recovery period from anesthesia is 2.77 μg/L in patients undergoing cervical spine surgery.

Objective To study the phenylpropanoid constituents from the roots of Euphorbia hylonoma Hand-Mazz. Methods The chemical constituents were isolated and purified by silica gel and Sephadex LH-20 column chromatography,and the structures were elucidated on the basis of chemical properties and spectral data. Results Three phenylpropanoid constituents were isolated from the acetone extracts of the roots of Euphorbia hylonoma Hand-Mazz,which were hexadecyl-3-methoxy-4-hydroxybenzeneacrylate Ⅰ,ethyl brevifolincarboxylate Ⅱ and(+) 3′-angeloyl-4′-isovalerylcis-khellactone peuformosin Ⅲ. Conclusion The above compounds were isolated from Euphorbia hylonoma Hand-Mazz for the first time,and the compound Ⅲ was the first obtained from the Euphorbiaceae.

BACKGROUND:Choosing internal fixator implants with good strength and stiffness is the key to repair femoral neck combined with ipsilateral subtrochanteric fractures. OBJECTIVE:To compare the biomechanical properties of different implant fixation for femoral neck combined with ipsilateral subtrochanteric fractures. METHODS:Totaly 24 adult antiseptic cadaver specimens were used to produce fracture models with femoral neck fracture combined with 5 cm of ipsilateral subtrochanteri medical cortical defect, and were divided into femoral proximal locking plate group, lengthening proximal femur anti-rotation intramedulary nail group and lengthening proximal femoral nail group according to the random number table method. The results of axial compression test, torsion test and axial compression failure rest in three groups were compared. RESULTS AND CONCLUSION: The axial compressive stiffness and failure load in lengthening proximal femur anti-rotation intramedulary nail group were significantly greater than those in femoral proximal locking plate group and lengthening proximal femoral nail group, and those in lengthening proximal femoral nail group were significantly greater than those in femoral proximal locking plate group (P < 0.05). The torsional stiffness in femoral proximal locking plate group was significantly greater than that in lengthening proximal femur anti-rotation intramedulary nail group and lengthening proximal femoral nail group, and that in lengthening proximal femur anti-rotation intramedulary nail group was significantly greater than that in lengthening proximal femoral nail group (P < 0.05). The indexes of biomechanical properties of specimens at the 4thand 8th weeks after fixation in three groups were slightly increased compared with those in 0 week after surgery, but the difference was no statisticaly significant (P > 0.05). These results demonstrate that to a certain extent, compared with the femoral proximal locking plate and lengthening lengthening proximal femoral nail, lengthening proximal femur anti-rotation intramedulary nail fixation for repair of femoral neck combined with ipsilateral subtrochanteric fractures has more biomechanical advantages.

Autogenous odontogenic materials are a new, highly biocompatible option for jaw restoration. The inorganic component of autogenous teeth acts as a scaffold to maintain the volume and enable donor cell attachment and proliferation; the organic component contains various growth factors that promote bone reconstruction and repair. The composition of dentin is similar to that of bone, which can be a rationale for promoting bone reconstruction. Recent advances have been made in the field of autogenous odontogenic materials, and studies have confirmed their safety and feasibility after successful clinical application. Autogenous odontogenic materials have unique characteristics compared with other bone-repair materials, such as the conventional autogenous, allogeneic, xenogeneic, and alloplastic bone substitutes. To encourage further research into odontogenic bone grafts, we compared the composition, osteogenesis, and development of autogenous odontogenic materials with those of other bone grafts. In conclusion, odontogenic bone grafts should be classified as a novel bone substitute.

Autogenous odontogenic materials are a new, highly biocompatible option for jaw restoration. The inorganic component of autogenous teeth acts as a scaffold to maintain the volume and enable donor cell attachment and proliferation; the organic component contains various growth factors that promote bone reconstruction and repair. The composition of dentin is similar to that of bone, which can be a rationale for promoting bone reconstruction. Recent advances have been made in the field of autogenous odontogenic materials, and studies have confirmed their safety and feasibility after successful clinical application. Autogenous odontogenic materials have unique characteristics compared with other bone-repair materials, such as the conventional autogenous, allogeneic, xenogeneic, and alloplastic bone substitutes. To encourage further research into odontogenic bone grafts, we compared the composition, osteogenesis, and development of autogenous odontogenic materials with those of other bone grafts. In conclusion, odontogenic bone grafts should be classified as a novel bone substitute.

In order to develop clinical diagnostic tools for rapid detection of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.
Antigens, Viral ; immunology ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; SARS Virus ; genetics ; immunology ; Yeasts ; genetics

【Objective】 To construct the Rb luciferase reporter gene assay system and detect the activation ability of Rb gene for screening the targeted drugs. 【Methods】 The synthetic Rb gene sequence was annealed to form a double-stranded DNA structure and then inserted into the polyclonal site of pGL6-TA. The junction product was transformed into E.coli DH5α competent cells for expanded culture, and the constructed pGL6-Rb-Luc plasmid and pGL6-TA plasmid were transfected into HEK293 cells. The monoclonal cell line HEK293-Rb-Luc with stable expression was screened by G418, and the activation and inhibition of Rb in HEK293-Rb-Luc were tested by serum and CDK4/6 inhibitor Palbociclib. 【Results】 The sequence of Rb reaction elements in pGL6-Rb-Luc was completely correct. The recovery of serum culture significantly increased the luciferase activity in HEK293-Rb-Luc (P<0.001). Compared with 0 nmol/L, 25, 50, 75 and 100 nmol/L, CDK4/6 inhibitor Palbociclib made the inhibition rate of Rb activity rise to 6.90%, 40.23%, 50.57% and 52.07%, respectively (P<0.05). 【Conclusion】 The Rb luciferase reporter gene detection system HEK293-Rb-Luc was successfully constructed, which can effectively detect the activation level of Rb.

【Objective】 To clone the full-length of human kidney and brain protein (KIBRA) coding sequence in eukaryotic expression vector and provide a model for studying the biological function of KIBRA in breast cancer cells. 【Methods】 Total RNA of human breast cancer cell line MCF7 was extracted. After reverse transcription, the full length of KIBRA (NM_001161661.2) coding region was amplified by PCR, and cloned into eukaryotic expression vector pCMV-Blank. After identification, it was defined officially as pCMV-KIBRA. Then it was transfected into MCF7 cells, and the expression of KIBRA was detected by real-time PCR and Western blotting after 48 hours. The primary, secondary and tertiary structures and post-transcriptional modification sites of KIBRA were analyzed with bioinformatics software. 【Results】 Bacterial PCR, double enzyme digestion and DNA sequencing results showed that the correct sequence of KIBRA was inserted into the vector pCMV-KIBRA. The mRNA and protein expressions of KIBRA were significantly increased in MCF7 cells transfected with pCMV-KIBRA. Bioinformatics analysis showed that KIBRA was composed of 1119 amino acids. There were 52 phosphorylation sites, 1 acetylation site and 5 ubiquitination sites, and the protein structure was mainly α-helix and random coil. 【Conclusion】 The eukaryotic expression vector of full-length of human KIBRA coding sequence was successfully constructed and overexpressed in breast cancer cell line MCF7, which can lay a foundation for studying the biological function of KIBRA in breast cancer.

【Objective】 To construct the eukaryotic expression vector carrying the human wild-type p27 and lacking nuclear localization signal p27△NLS coding sequences， and the express them in HEK293T cells， which may contribute to investigating the different locations and roles of p27 in the cytoplasm and nucleus. 【Methods】 Total RNA was prepared from human breast cancer MCF7 cells， and cDNA was obtained by reverse transcription-polymerase chain reaction （RT-PCR）. After amplification of the p27 CDs and non-NLS fragments by PCR， full length p27WT （CDKN1B， NM_004064.5） and p27△NLS coding regions were obtained. PCR products were then subcloned into the eukaryotic expression vector pCMV-Blank. After identification with bacterial PCR， double restriction enzyme digestion and sequencing， they were defined officially as pCMV-p27WT and pCMV-p27△NLS， respectively. The recombinant plasmids were transfected into HEK293T cells by electroporation. After 48 h， the levels of p27 protein in the cytoplasm and nucleus were detected by Western blotting. 【Results】 The sequencing results showed that the sequences of p27WT and p27△NLS inserted into the plasmids were both correctly consistent with that of NM_004064.5. After transfection with pCMV-p27WT， total p27 protein expression was increased and distributed in both the cytoplasm and nucleus of HEK293T cells. After transfection with pCMV-p27△NLS， p27 protein was significantly increased and almost entirely localized in the cytoplasm of HEK293T cells. 【Conclusion】 The eukaryotic expression plasmids of human p27WT and p27△NLS coding sequences were successfully constructed and overexpressed in HEK293T cells. This research may lay a foundation for investigating the biological function of p27 in the cell cycle progression of tumor cells.