Objective: To construct the eukaryotic expression plasmid carrying hTERT-P2A-EGFP, and to explore its expression and transfection efficiency in the HEK293FT cells. Methods: The recombinant plasmid was constructed by using pBABE-puro-hTERT and pRRLSIN-cPPT-MSCV-EGFP plasmids. The hTERT,P2A,and EGFP genes were obtained using pBABE-puro-hTERT as template by PCR. And the correct hTERT was inserted into pRRLSIN-cPPT-MSCV-EGFP vector. Then the recombinant plasmid containing hTERT-P2A-EGFP gene was obtained and identified. The HEK293FT cells were transfected by the recombinant plasmid, and the expression of green fluorescence protein(GFP) was observed by fluorescence microscope. Results: The PCR results showed that the fragments of hTERT, P2A, and EGFP were 3 400, 110 and 720 bp. And the length of gene fragment(hTERT-P2A-EGFP)was 4 300 bp by enzyme digestion. The results of sequencing showed that the 1 547 site of the target gene was mutated. Using site-directed mutagenesis, the 1 547 site was successfully mutated. And the target gene sequence was completely identical with the sequence published in GenBank. The recombinant plasmid was transfected into the HEK293FT cells, and GFP was observed in the cells.The results of flow cytometry showed that the transfection efficiency of recombinant plasmid was 44.8%. Conclusion: The recombinant plasmid carrying hTERT-P2A-EGFP gene is successfully constructed, and it can be used for cell transfection.

Objective: To study the inhibitory effect of retinoblastoma 94(Rb94) gene combined with radiotherapy ionizing radiation on the growth of esophageal carcinoma cells of tumor-bearing nude mice, and to clarify the synergistic effect of Rb94 gene and radiotherapy in inhibiting the growth of tumor cells. Methods: The models of tumor-bearing BALB/c-nu nude mice were built by inoculating the K150 cells. The model mice were divided into five groups:blank control(no any treatment), Ad-LacZ(control adenovirus including LacZ gene but not Rb94 gene, Ad-LacZ was transfered into tumor xenograft on 0, 3, 7 d separately), Ad-Rb94(tumor xenograft was transfected with Ad-Rb94 on 0, 3, 7 d separately), radiation (tumor xenograft was irradiated with 4 Gy γ-radiation on 1, 4, 8 d separately) and Ad-Rb94 combined with radiation(combination group, tumor xenograft was irradiated with 4 Gy γ-radiation after transfected with Ad-Rb94) groups. The volumes and the weights of esophageal carcinoma and the inhibitory rates of tumor growth of the mice in various groups were detected. The expression levels of ABL and JNK kinase in tumor tissue of the mice in various groups were measured,and the pathological changes of tumor tissue were investigated. Results: The speeds of tumor growth of the nude mice in Ad-RB94, radiation, and combination groups were slower than that in control group. The volume of esophageal carcinoma in combination group at day 15 after treatment was markedly smaller than those in Ad-RB94 and radiation groups,and there were significant differences compared with control group and Ad-LacZ group (F=26.7,23.8;P<0.01). The tumor weight of the nude mice in combination group was the lightest at the end of treatment; the inhibitory rate of tumor growth in combination group reached 81.16% and was significantly higher than those in Ad-Rb94 group(57.84%)and radiation group(38.20%)(P<0.01). The expression levels of ALB and JNK kinase in tumor tissue of the mice in combination group was markedly higher than those in control group(P<0.01). Compared with other groups, the tumor cells in combination group had fewer karyokinesis and lower level of nuclei hyperchromasia. Conclusion: Rb94 gene combined with radiotherapy shows synergistic effect in inhibiting the growth of tumor of tumor-bearing nude mice.

Objective: To investigate the relationship between visfatin and insulin resistance (IR)of type 2 diabetes mellitus(T2DM) through the classic insulin signaling pathway phosphatidy inositol-3 kinase (PI3K). Methods: The human T2DM preadipocyte cells were recoveried, extended and differentiated. The visfatin overexpression vectors were built, transformated, cultivated and extracted. The fat cells were transfected with different overexpression levels (0.0, 1.0, 2.5 and 5.0 μg); 0.0 μg group was used as control group,and the remaining three groups as observation groups. The mRNA expression levels of visfatin, insulin receptor substrate-1(IRS-1), insulin receptor substrate-2(IRS-2) and PI3K (P85α) were detected by Q-PCR. The protein expression levels of visfatin, IRS-1, IRS-2, PI3K (P85α), IRS-1 and IRS-2 phosphorylation levels were measured by Western blotting method.The glucose uptake rates of fat cells were determined by [³H]-2-deoxidation-D-glucose uptake assay. Results: The expression levels of visfatin mRNA and protein in various groups were increased with the increase of transfection concentration gradient (P<0.01); and the constructed overexpression vector of visfatin was effective. With the increase of visfatin expression, the mRNA and protein expression levels of IRS-1 and PI3K (P85α) and IRS-1 phosphorylation degrees in various groups were increased (P<0.01), but the IRS-2 mRNA and protein expression levels had no obvious changes(P>0.05).The glucose uptake rates of fat cells were elevated with the increasing of visfatin expression (P<0.05). Conclusion: The visfatin overexpression of fat cells in vitro can increase the expression levels of IRS-1 and PI3K.

Objective: To study the effects of SiO₂ nanoparticles on the organs of mice in vivo after intratracheal instillation, and to provide the basis for safety evaluation of SiO₂ nanoparticles. Methods: Forty female BALB/c mice aged 6-8 weeks were randomly divided into control group (saline), low dose of SiO₂ group (7 mg·kg^-1), middle dose of SiO₂ group (21 mg·kg^-1), and high dose of SiO₂ group (35 mg·kg^-1). 1 and 15 d after five times of non-exposed intratracheal instilation infection (once every 3 d), the mice were sacrificed and the left lungs,the right kidneys, livers, hearts and spleens were collected and embedded in paraffin. The morphology of tissue sections was observed under light microscope after hematoxylin-eosin (HE) staining. The eyeball blood was obtained and the biochemical indicators of liver and kidndy functions were detected. Results: Compared with control group, there were alveolar interval thickening, inflammatory cell infiltration, and a small amount of small arterial thrombosis in the lungs; granulomatous inflammatory cell infiltration and a small amount of focal necrosis of liver cells in the livers; red pulp enlargement, hyperemia, and more visibly scattered megakaryocytes in the spleens in SiO₂ nanoparticles groups in a dose-dependent manner, especially in middle and high doses of SiO₂ groups. After 15 d of injection, the damages alleviated with the prolongation of time. There was some inflammatory cell infiltration in the kidney tissue of the mice in SiO₂ nanoparticle groups. The biochemical indicator detection results showed that alanine aminotransferase(ALT) and aspartate transaminase(AST) levels in SiO₂ nanoparticles groups varied, suggesting the liver cell damages were at different degrees; the changes of urea nitrogen(BUN) and creatinine(Cr) levels in SiO₂ nanoparticle groups remindered the kidney function alteration, but there were no obvious dose- and time- dependent effects. Conclusion: Intratracheal instillation of SiO₂ nanoparticles can influence the major organs of the mice and mainly displays in the inflammation and injuries in the lung, liver, and spleen.

Objective: To investigate the effects of ursolic acid (UA) on the proliferation and differentiation of osteoclasts (OC), and to explore its role in orthodontic force-induced root resorption and its relationship with OC. Methods: The mononuclear/macrophage cells RAW264.7 were induced to the OC.Tacrolimus acid phosphatase (TRAP) staining and bone resorption lacunae observation were used to identify the induction. CCK-8 method was used to select the appropriate concentration of UA for RAW264.7 cell-free biotoxicity and to observe its effect on the proliferation and differentiation of RAW264.7.In experimental groups, UA with gradient concentrations (1.0,2.5,5.0,10.0,20.0and 40.0 μmol·L^-1)were added.UA was not added in control group. Results: The TRAP staining and bone resorption lacunae observation showed that after the RAW264.7 cells were induced for 5 d, the TRAP staining positive cells were found; the resorption lacunae were rounded,and oval, etc,the bottom wall was coarser,and the boundary was clear,which indicated that the RAW264.7 cells were successfully differentiated into the osteoclasts.The CCK-8 detection results showed that high concentration of UA (> 10.0 μmol·L^-1) significantly inhibited the proliferation of OC;the appropriate concentration of UA (5.0 μmol·L^-1) was in the biological safety concentration range and could inhibit the OC proliferation; low concentration of UA (<2.5 μmol·L^-1) had no effect. Conclusion: RANKL can induce the differentiation and maturation of RAW264.7 cells. UA is correlated with the proliferation and differentiation of OC; UA has inhibitory effect on OC at the appropriate concentration (5.0 μmol·L^-1) in a time-dependent manner.

Objective: To study the expression characteristics of Dickkopf1 (DKK1) in different time and space during tooth development of the postnatal mice, and to provide the theoretical basis for clarifying the mechanism of Wnt signaling pathway in regulating the tooth development. Methods: The postnatal Kunming mice at days 0.5, 6.5, 12.5, 18.5, 24.5, and 30.5 respectively after birth were selected and divided into various groups by time,three in each group.The mice in each group were sacrificed and the paraffin sections of mandibular bone including the first molar were prepared at the thickness of 5 μm, followed by HE staining and immunohistochemical staining in order to detect the expressions of DKK1 in tooth tissue and periodontal tissue. Results: At 0.5 d after birth, the mandibular first molar tooth germ was in the bell stage. At 6.5 d the enamel development of mandibular first molar was almost completed, and the epithelium root sheath extended to the root direction. At 12.5 d the dentin development of crown was completed, with the root formatted about 1/3. At 18.5 d the root had formatted about 2/3. At 24.5 d the root had reached the full length. At 30.5 d the apical foramen was narrow, and the root development was basically completed. There was no DKK1 expression at 0.5 d, but it expressed in the odontoblasts and predentin at 6.5 d. From days 12.5 to 30.5,the expressions of DKK1 were positive in periodontal ligament, alveolar bone, and cellular cementum as odontoblasts, which were gradually increased with the prolongation of time. However, no expression of DKK1 was detected in the pulp. Conclusion: DKK1 shows regular expressions at different tooth developmental stages after birth, suggesting its potential role in the growth of dentin and periodontal tissues.

Objective: To explore the influence of fermented red ginseng extract (FRGE) in the proliferation of rat glomerular mesangial cells (GMCs) and the degradation of extracellular matrix(ECM)under high sugar stimulation, and to clarify the prevention and treatment effects of FRGE on diabetic nephropathy ( DN) and the possible mechanism. Methods: The rat GMCs were cultured and divided into normal concentration of D-glucose (NG) group, high concentration of D-glucose (HG) group and high concentration of D-glucose plus different concentrations (3.75, 7.50, 15.00 mg·L^-1) of FRGE groups. The proliferation rates of rat GMCs were detected with MTT,and the type Ⅳcollagen(Col Ⅳ) levels in supernatants of the GMCs were detected by ELISA. The protein expressions levels of matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) were detected with Western blotting method. Results: Compared with NG group, the proliferation rate of GMCs in HG group was increased(P<0.01), the Col Ⅳ level was increased(P<0.01),the MMP-2 expression level was decreased, and the TIMP-2 expression level was up-regulated(P<0.01). Compared with HG group, the proliferation rates of GMCs in various FRGE groups were decreased(P<0.01), the Col Ⅳ levels were decreased(P<0.01),the expression levels of TIMP-2 were reduced(P<0.01),and the expression levels of MMP-2 were increased(P<0.01). Conclusion: FRGE can inhibit the proliferation of rat GMCs induced by high sugar and promote the ECM degradation to delay the occurrence and development of DN.

Objective: To investigate the influence of multilayer alginate chitosan sustained-release microspheres loading vascular endothelial growth factor(VEGF) and vancomycin in the proliferation and osteogenic differentiation of human plaacenta-derived mesen chymal stem cells(HPMSCs), and to provide theoretical basis for its clinical application in the repair of bone defect. Methods: The microspheres were prepared based on the previous research and HPMSCs were co-cultured with drug (VEGF/vancomycin)-loaded microspheres (drug-loaded microspheres+HPMSCs group), non-drug loaded microspheres (microspheres+HPMSCs group) and without any microspheres (HPMSCs group). Then the proliferation rate of HPMSCs was identified by CCK-8 kit. The osteogenic differentiation potential of HPMSCs was detected by Alizarin red staining and alkaline phosphatase (ALP) kit when the HPMSCs had been co-cultured with drug loaded microspheres in osteogenic medium (HPMSCs+ drug-loaded microspheres+induction group), non-drug loaded microspheres in osteogenic medium (HPMSCs+microspheres+induction group), without any microspheres in osteogenic medium (HPMSCs+induction group) and without any micropheres in normal medium (HPMSCs+PBS group) for 21 d. Results: Compared with HPMSCs group,the proliferation rates of HPMSCs in drug-loaded microspheres+HPMSCs and microspheres+HPMSCs groups had no significant changes (P>0.05).The calcium deposition in HPMSCs+drug-loaded microspheres+induction group was more than those in microspheress+HPMSCs+indution group and HPMSCs+induction group after Aalizarin red staining; the ALP activity in drug-loaded microspheres+HPMSCs+indution group was higher than those in microspheres+HPMSCs+indution group and HPMSCs+induction group (P<0.05),and the ALP activity in microspheres+HPMSCs+induction group was higher than that in HPMSCs+induction group(P<0.05). Conclusion: The sustained-release microspheres loading VEGF and vancomycin have no significant effect on the proliferation activity of HPMSCs and the microspheres could stimulate the osteogenic differentiation of HPMSCs.

Objective: To study the influence of metformin and paclitaxel in the proliferation and apoptosis of human ovarian cancer SKOV3 cells in vitro, and to clarify the synergistic effect of metformin and paclitaxel. Methods: The ovarian cancer SKOV3 cells were divided into blank control group, different concentrations (0.01, 0.50, 1.00, 5.00, 10.00mmol·L^-1) of metformin groups and combined treatment groups(metformin combined with paclitaxel with different concentrations). The inhibitory rates of proliferation of SKOV3 cells after treated with different concentrations of metformin were detected by MTT method. The apoptotic rates of SKOV3 cells after treated with different concentrations of metformin were measured by flow cytometry. The inhibitory rates of proliferation of SKOV3 cells after treated with metformin and paclitaxel were determined by MTT method. Results: The MTT results showed that the inhibitory rates of proliferation of SKOV3 cells in different concentrations of metformin groups were increased in concentration-and time- dependent manner; there were significant differences compared with blank control group (P<0.05). The flow cytometry results showed that the apoptotic rates of SKOV3 cells in different concentrations of metformin groups were increased; compared with control group, with the increasing of concentrations of metformin, the apoptotic rates of SKOV3 cells in experimental groups 48 h after treatment were increased significantly (P<0.05); the percentages of SKOV3 cells in G₀/G₁ phase were decreased with the increasing of metformin concentrations(P<0.05) and the percentages of SKOV3 cells in G₂/M phase were increased(P<0.05). The MTT results showed that the inhibitory rate of proliferation of SKOV3 cells in 1.00 mmol·L^-1 metformin+paclitaxel group was higher than that in 0.50 mmol·L^-1 metformin+paclitaxel group was higher than that in 0.50 mmol·L^-1 metformin+paclitaxel group(P<0.05),and the inhibitory rates of proliferation of SKOV3 cells in combined treatment groups were higher than those in paclitaxel alone groups(P<0.05). Conclusion: Metformin inhibits the proliferation of ovarian cancer SKOV3 cells through the induction of apoptosis. Metformin can enhance the cell proliferation inhibition of paclitaxel on ovarian cancer SKOV3 cells. The combination of metformin and paclitaxel has a synergistic reaction on SKOV3 cells.

Objective: To investigate the influence of magnetic Fe₃O₄ nanoparticles in the expressions of Caveolin-1 and Clathrin Heavy Chain proteins in organ tissues of the rats, and to clarify its mechanism. Methods: Twenty-four Wistar rats were randomly divided into control group, and low,medium and high doses of magnetic Fe₃O₄ nanoparticle groups by weights. 24 h after tail vein injection of different doses of magnetic Fe₃O₄ nanoparticles, the organ tissues were obtained. Western blotting method was used to detect the expressions of Caveolin-1 and Clathrin Heavy Chain proteins in the main organ tissues of the rats. Real-time fluorescent quantitative PCR was used to measure the expressions of Caveolin-1 and Clathrin Heavy Chain mRNA. Results: Compared with control group,the expressions levels Clathrin Heavy Chain protein and mRNA in liver and spleen tissues of the rats in medium and high doses groups were significantly increased (P<0.05).The expression level of Clathrin Heavy Chain mRNA in kidney tissue in high dose group was higher than those in other groups(P<0.05). The Caveolin-1 expression levels in the organ tissues of rats had no statistical differences between various groups (P>0.05).Compared with control group, the Caveolin-1 mRNA expression levels in liver, spleen, and lung tissues of the rats in low,medium and high doses groups were significantly increased (P<0.05);there were no statistical differences in the expression levels in kidney tissue of the rats between various groups(P>0.05). Conclusion: Magnetic Fe₃O₄ nanoparticles could enhance the expressions of Clathrin Heavy Chain in the liver, spleen, and lung tissues of the rats. Endocytosis of Clathrin Heavy Chain protein is one way for magnetic Fe₃O₄ nanoparticles into the liver, lung, spleen cells of the rats.

Objective: To construct the eukaryotic expression vectorsof human Aurora A(AURKA), and to detect the expression and purification of AURKA in Pichia pastoris X33 and MCF-7 cells. Methods: The AURKA was amplified by polymerase chain reaction (PCR). The genes were digested with XhoⅠ and XbaⅠ, and respectively ligated into pPICZαA and pcDNA3.1(+) to construct the recombinant plasmids pPICZαA- AURKA and pcDNA3.1(+)-AURKA which were transfected into Pichia pastoris X33 and MCF-7 cells by Gene Pulser Xcell^TM Electroporation System. The expressions of AURKA protein were detected by SDS-PAGE and Western blotting methods after screening in Pichia pastoris X33 and MCF-7 cells, respectively. The purification of proteins was determined using Ni-NTA column. The activity of AURKA was analyzed by autoradiography. The proliferation of MCF-7 cells after transfected with AURKA was detected by cell counting method. Results: The 1 212 bp genes were amplified from the pPICZαA-AURKA and pcDNA3.1(+)-AURKA by PCR, which indicated that two eukaryotic expression vectors were successfully constructed. The relative molecular mass of recombinant AURKA protein was approximately 50 000. The recombinant AURKA phosphorylated the histone H3 by autoradiography, which manifested that it had activity. Compared with the cells transfected with empty plasmid and WT MCF-7 cells, the number of alive MCF-7 cells was significantly increased after transfected with AURKA for 24 and 48 h. Conclusion: AURKA is successfully expressed and purified in Pichia pastoris X33 and MCF-7 cells. The over-expression of AURKA can promote the proliferation of MCF-7 cells

Objective: To investigate the influence of P27RF-Rho mRNA gene silencing in the drug sensitivity of 5-fluorouracil(5-Fu)to the liver cancer SMMC cell line,and to provide theoretical basis for the treatment of advanced liver cancer. Methods: The P27RF-Rho RNAi vector was constructed and the P27RF-Rho gene silencing lentivirus were used to infect the SMMC7721 cells. Western blotting method was used to detect the gene silencing effect. The SMMC7721 cells were divided into Scramble-siRNA group, 5-Fu group, P27RF-Rho siRNA group and P27RF-Rho siRNA + 5-Fu group. Western blotting was used to detect the transfection efficiency of RNAi. MTT method was used to detect the cell growth in various groups. Scratching test was used to detect the migration ability of cells in various groups. Transwell experiment were used to detect the invasion ability of cells in various groups. The expressions of P27 and RhoC protein were detected by Western blotting method. Results: P27RF-Rho RNAi lentiviral vector was successfully constructed. The Western blotting results showed that the expression of P27RF-Rho protein in P27RF-Rho siRNA group was decreased compared with 5-Fu group and Scramble-siRNA group(P<0.05). Compared with other three groups, the growth speed of the cells in P27RF-Rho siRNA + 5-Fu group was significantly decreased(P<0.05). The migration ability of the cells in P27RF-Rho siRNA + 5-Fu group was significantly lower than those in other three groups (P<0.01);the average number of cells passing through the Transwell microporous membrane was significantly less than those in other three groups (P<0.01).The Western blotting analysis results showed that the expression level of P27 protein in the cells in P27RF-Rho siRNA + 5-Fu group was significantly higher than those in other three groups(P<0.05);the expression level of RhoC protein was significantly lower than those in other three groups(P<0.05). Conclusion: P27RF-Rho gene silencing can significantly enhance the drug sensitivity of 5-Fu to SMMC7721 cells.

Objective: To observe the repair effect of nano hydroxyapatite/magnesium(nHA-Mg)porous composite material modified by poly lactic acid/glycolic acid copolymer(PLGA) on the jaw bone defect of rabbits,and to elucidate the mechanisms preliminary. Methods: The mandibular defect models of 10 mm×5 mm×1 mm in the 18rabbits were established.Nine rabbits were selected and implanted with nHA-Mg composite materials modified by PLGA in the left as experimental group and the right as blank control group;the other nine were implanted with nHA-Mg in the left as positive control group and the right as blank control group.The rabbits were sacrificed at 4, 8, 12 weeks (respectively 3 of experimental group and positive control group every time)and the mandibular defect areas were intercepted and observed by imaging and histological examination;the sizes of the newborn trabecula area and residual materials in experimental group and positive control group were compared. Results: Compared with positive control group and blank control group, the percentage of newborn trabecula area of the rabbits in experimental group was increased (P<0.05),and there was no statistic difference between positive control group and blank control group (P>0.05).The imaging results showed that the new bone formation can be observed in experimental group and better than positive control group and blank control group.The paraffin section results indicated that there were visible vascular tissue and newborn trabecula,the osteoblasts gathered around the bone trabecula; a lot of pits were located in the bone trabecula, and the pits contained osteocytes in experimental group.As the prolongation of time,thickened newborn trabecula, dense arrangement and trend of converting to lamellar bone were observed in experimental group.The hard tissue section results showed that the remaining amount of materials in experimental group was more than that in positive control group. Conclusion: nHA-Mg porous composite materials modified by PLGA can effectively reduce the rate of degradation in the body, promote osteogenesis and guide the bone regeneration.

Objective: To investigate the synergistic killer effect of natural killer cells(NK cells) combined with tamoxifen(TAM) on breast cancer cells(BCC)through the experiment in vitro,and to explore its mechanism. Methods: Three kinds of BCC with different receptor expression levels were selected for the experiment. Blank control group, different concentrations of TAM groups and different time groups were setup.MTT assay was used to detect the inhibitory rates of proliferation of cells,and the final experiment concentration of 5 μmol·L^-1 was determined.The cells were divided into natural-release group,largest-release group,TAM group,NK cells group, and combined-experimental group(BCC+NK cells+TAM),and the synergistic killer effect of NK cells combined with TAM in different effector-target ratios were detected with Calcein-AM release assay.In ELISA assay the cells were divided into blank control group (NK cells),NK cells+TAM group, NK cells+BCC group and combined-experimental group,and the levels of IFN-γ and TNF-α in the NK cells in various groups were measured.In flow cytometry detection the cells were divided into blank control group (NK cells),NK cells+TAM group,NK cells+ BCC group, and combined-experimental group,and the expression levels of NKp46,CD158a,CD158b,CD158b2,and CD158e were determined;while the cells were divided into blank control group (BCC),BCC+TAM group,BCC + NK cells group, and combined-experimental group,and the expression levels of the MICA,ULBP1 and ULBP2 were detected. Results: The MTT assay results showed that the inhibitory rates of proliferation of 3 kinds of BCC had obvious time- and concentration-dependence (P<0.05).The Calcein-AM release assay results showed that the killing-rates of BCC in TAM groups were increased with the increase effector-target ratios of compared with NK cells group;and the killing-rate in combined experimental group was obviously higher than those in NK cells and TAM groups(P<0.05).The ELISA assay results showed that the levels of TNF-α and IFN-γ of NK cells in various experimental groups with BCC or not were increased compared with blank control group(P<0.05 or P<0.01);the levels were significantly increased when combined with TAM (P<0.05).The flow cytometry results showed that the NKp46 expression levels in various experimental groups were elevated compared with blank control group(P<0.05);the expression levels of CD158a, CD158b,CD158b2, and CD158e were significantly decreased(P<0.05);the expression levels MICA,ULBP1, and ULBP2 in BCC were significantly increased (P<0.05). Conclusion: The NK cells combined with TAM has the synergistic killer effect on the BCC in vitro.The synergetic mechanism may be as follows:TAM could increase the secretion of TNF-α and IFN-γ of NK cells to enhance their cytotoxicity;TAM also could up-regulate the expression levels of activating receptors and activating ligands,and down-regulate the expression levels of inhibitory receptors to increase the killing ability of NK cells.

Objective: To explore the effect of tectorigenin on myocardial fibrosis(MF) in the rats and clarify the related mechanism,and to provide reference for its clinical application. Methods: Sixty Wistar rats were randomly divided into normal control group,model group,positive drug (capropril) control group, and low,middle,high doses of tectorigenin groups(n=10).Except normal control group, the rats in other groups were used to construct MF models by subcutaneous injection of 5 mg·kg ^-1·d ^-1 isoproterenol (Iso) for 7 d.The rats in tectorigenin groups and captopril group were intragastricly administrated with different doses of tectorigenin (25,50,100 mg·kg^-1·d^-1)and captopril(10 mg·kg ^-1·d ^-1) from the second day after modeling for consecutive 28 d.Bl-420E+ biological function experiment system was used to detect the heart function; Heart mass index (HMI) and left ventricular mass index (LVMI) were measured after experiment.UV detection was used to measure the levels of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) in myocardial tissue.Microplate reader was used to measure the activities of lactic dehydrogenase(LDH) and creatine kinase(CK) and the levels of nitric oxide (NO)in serum.ELISA were used to detect the levels of collagen typeⅠ(ColⅠ) and collagen type Ⅲ (Col Ⅲ) in myocardium tissue of the rats.The pathological changes of myocardium tissue of the rats in various groups were observed by HE staining. Results: Compared with normal control group,the HR of rats in model groups was increased,and the left ventricular systolic pressure (LVSP) was decreased(P<0.01); the HMI and LVMI were increased(P<0.05), the levels of MDA in left ventricular myocardial tissue was increased(P<0.01), and the activity of SOD was decreased,the levels of serum ColⅠ,Col Ⅲ and the activities of LDH, CK were also increased(P<0.01);the level of NO in serum was decreased(P<0.01).Compared with model groups, the HR were decreased,LVSP were increased, and HMI and LVMI of the rats in different doses of tectorigenin groups were decreased in a dose-dependent manner; the levels of MDA were reduced;the activities of SOD were increased in myocardium tissue,and the CK activities and the ColⅠ and ColⅢ levels were decreased(P<0.05 or P<0.01);the LDH activities in middle and high doses of tectorgenin groups were decreased(P<0.01); and the levels of NO in serum in different doses of tectorigenin groups were significantlyincreased(P<0.05 or P<0.01). Conclusion: Tectorigenin could inhibit the MF induced by Iso in the rats, and its mechanism may be related to antioxidation,scavenging free radical and inhibition of collagen synthesis.

Objective: To observe the protective effect of deproteinized extract of calf blood (DECB) on the kidney injury ofthe diabetic rats, and to discuss its mechanism preliminarily. Methods: The male Wistar rats were intraperitoneally injected with STZ (65 mg·kg^-1) to establish the diabetes models, then the model rats were randomly divided into model (M)group, and metformin (MMet, 105 mg·kg^-1) group, low dose of combined administration (ML,105 mg·k^-1 metformin+94.5 mg·kg^-1,DECB) group, medium dose of combined administration(MM, 105 mg·kg^-1 metformin+ 189 mg·kg^-1 DECB) group, high dose of combined administration(MH, 105mg·kg^-1 metformin +378 mg·kg^-1,DECB) group, anotherten Wistar rats were selected as normal control(NC)group.The rats were intragastrically administed with metformin and intraperitoneally injected with DECB, once a day, total of 8 weeks.The rats in NC group and M group were given normal saline solution.The weights and blood glucose levels of the rats in various groups were determined;the levels of serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), blood urea nitrogen (BUN), urinary albumin (UAlb), urine creatinine (UCR), serum creatinine (SCr), total cholesterol (TC), triglyceride (TG), uric acid (UA), glutathione (GSH), and malondialdehyde (MDA) were detected; the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined.The pathological changes of kidney tissue of the rats in various groups were detected by HE staining. Results: Compared with NC group, the weight of the rats in M group was reduced(P<0.05),and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG,and MDA were increased(P<0.05);the levels of HDL-C and GSH were obviously reduced(P<0.05),and the activities of SOD adn GSH-Px were obviously reduced(P<0.05).Compared with M group,the weights of the rats in MM and MH groups were increased (P<0.05), and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG, and MDA were decreased (P< 0.05);the levels of HDL-C and GSH were increased (P<0.05),and the activities of SOD and GSH-Px were increased (P<0.05).The pathological observation of kidney tissue showed that the rats in M group had obvios kidney tissue lesions with glomerular congestion and renal tubular edema compared with NC group; compared with M group,the pathological changes of the kidney tissue of the rats in drug administration groups were significantly improved, the renal tubular vacuoles were reduced, and the interstitial hyperplasia was not obvious. Conclusion: DECB combined with metformin can reduce the blood glucose level, regulate blood lipid, improve the pathological changes of kidney tissue in the diabetic rats, reduce the renal damage, and enhance the antioxidant capacity of the body.

Objective: To evaluate the clinical efficacy of treatment of sudden deafness using hyperbaric oxygenation, and to provide guidance for its treatment. Methods: The studies about sudden deafness treated with hyperbaric oxygenation combined with drug were retrieved in Cochrane Library, PubMed, MEDLINE, Wanfang Database and CNKI up to July 2016. RavMan 5.3 provided by Cochrane was used for Meta-analysis. Results: Thirty-nine studies which were randomized controlled trials (RCTs)were included, a total of 4 599 cases,of which 2 418 cases received hyperbaric oxygenation therapy combined with drug treatment as treatment group and 2 181 cases received drug treatment alone as control group. A total of 36 studies about the comparison of clinical efficacy between treatment group and control group were included in the Meta-analysis, the result showed that treatment group had a significant clinical efficacy than control group(RR:1.26,95%CI:1.22-1.30,P< 0.0001). A total of 6 studies about the improvement of the pure tone average(PTA)in the two groups were included in the Meta-analysis, the PTA gain was significant better in hyperbaric oxygenation group(WMD:11.8,95%CI:4.77-18.83,P< 0.000 1).A total of 11 studies about the stage of using hyperbaric oxygenation therapy were included in the Meta-analysis, the result suggested that the early use of hyperbaric oxygenation therapy had better effect than the late use of hyperbaric oxygenation (RR:1.42,95%CI:1.31-1.53,P< 0.000 01). Conclusion: The clinical efficacy of hyperbaric oxygenation combined with drug therapy is better than drug therapy alone, the PTA gain is significantly improved and the introduction of hyperbaric oxygenation therapy at the earlier stage shows better results.

Objective: To evaluate the relationship between the phthalate ester exposure and the population obesity with Meta-analysis, and to provide a new idea for prevention and control of obesity. Methods: A comprehensive search was performed in English databases (Pubmed, Web of Science, The Cochrane Library, Elsevier Science Direct and OVID) and Chinese databases (Sinomed database, CNKI database, VIP database, Wanfang database).The studies about the relationship between phthalate ester exposure and the population obesity were retriveded.The Chinese and English studies were selected according to the inclusion criteria and exclusion criteria.Meta-analysis was performed using RevMan 5.3 software. Results: Six studies were finally obtained, involving 1259 samples.The Meta-analysis results showed that the monobutyl phthalate (MBP) level in urine of the obesity population was increased 4.1 times compared with the normal population (95%CI:1.43-6.76); while the combined effect values of dibutyl phthalate (DBP), di-2-ethylhexyl phthalate (DEHP) and diethyl phthalate (DEP) level in serum of the population in two groups were 1.17 (95%CI:0.64-1.69), 0.80 (95%CI:0.13-1.48), and 0.72 (95%CI:-0.19-1.63);the combined effect values of monoethylhexyl phthalate (MEHP) and monoethyl phthalate (MEP) levels in urine were 1.75 (95%CI:-0.45-3.96) and 2.75 (95%CI:0.36-5.15); there were no significant differences (P >0.05). Conclusion: The elevated MBP levels in the urine may be a risk factor for obesity in the population, suggesting that MBP may contribute to obesity.

Objective: To clarify the significance of the fractional anisotropy(FA) value and apparent diffusion coefficient(ADC) value of diffusion tensor imaging(DTI) applied in the glioma grading with Meta-analysis, and to provide a reference for selecting the treatment program and judging the prognosis of glioma. Methods: Computer search was performed in PubMed database, EMBASE database,CBM, CNKI, Wanfang database, and VIP database.The relevant neurosurgery magazines and neuroimaging magazines, the proceedings of academic meeting were retrieved by hand and the references of related reviews were retrospectively retrieved.Then the case-series studies according to inclusion and exclusion criteria were screened,and the qualities of included studies were evaluated.Meta-analysis was carried out by Review Manager 5.2 software. Results: Nine studies were included.The FA value of low-grade glioma(LGG) in tumor centre was lower than that of high-grade glioma(HGG) (χ²=11.94,df=8,P=0.15;I²=33%,Z=2.78,P=0.005;WMD_FA=-0.02,95%CI:-0.03--0.01),and the ADC value of LGG in tumor centre was higher than that of HGG (χ²=5.26,df=4,P=0.26;I²=24%,Z=2.19,P=0.03;WMD_ADC=0.15,95%CI:0.02-0.28). Conclusion: When the FA value in tumor centre of the glioma is increasd,the ADC value is decreased,and the grade of glioma is increasd. The FA value and ADC value of DTI can help determine the grade of gliomas.

Objective: To compare the effectiveness and safety of Dengzhanhuasu and Danshen Injection in the treatment of the patients with angina pectoris of coronary heart disease with Meta-analysis, and to provide basis for their application in clinic. Methods: Computer search was performed using Cochrane Central Register of Controlled Trial,Ovid-medline,EMBASE,PubMed,CNKI,Wanfang,VIP and CBM,and the literatures about the effectiveness and safety of Dengzhanhuasu and Dansheng Injection in the treatment of the patients with angina pectonis of coronary heart disease were obtained. According to the principle of selection and elimination, the literatures were selected, and the qualities of literatures were evaluated. The related data were extracted from the literatures.RevMan 5.3 software was used to extract the data for Meta-analysis. Results: A total of 167 literatures were retrieved, and 8 randomlized-controlled studies were selected according to the selection criteria. The evaluation results of the effectiveness of angina pectoris according to symptom showed that χ²=2.62,df=7,P=0.92,Z=6.85 (P<0.000 01), 95%CI:2.73-6.10. The evaluation of the effectiveness of angina pectoris according to electrocardiogram,χ²=6.37,df=7,P=0.50,Z=6.37(P=0.000 01),95%CI:1.94-3.49.The results of Meta-analysis showed that there were significant differences in the relief of symptoms and improvement of the electrocardiogram efficacy between Dengzhanhuasu and Danshen Injection in the treatment of angina pectoris of coronary heart disease (P<0.05). Conclusion: Dengzhanhuasu has better effectiveness in the treatment of the patients with unstable angina than Dansheng Injection and they have the same safety.

Objective: To explore the expressions of miRNAs related to accelerating senescence in serum of the patients with amnestic mild cognitive impairment (aMCI), and to clarify their effects in the pathogenesis of aMIC. Methods: The levels of miRNAs related to accelerating senescence (miR-132, miR-193b, miR-130b, miR-20a, miR-296, miR-329 and miR-206) were measured in the serum of the patients with aMCI (aMCI group,n=66) and healthy controls(control group,n=76) using quantitative real-time PCR(qRT-PCR). The genes targeted by the altered miRNAs were predicted by TargetScan 6.0. DAVID was used to analyze the function of miRNA target genes. The serum levels of brain derived neurotrophic factor (BDNF) and silent in formation regulator 1(SIRT1) were measured by enzyme-linked immunosorbent assay (ELISA) method. Results: The expression levels of miR-206 and miR-132 in serum of the patients in aMCI group were significantly higher than those in control group (P<0.05). BDNF and SIRT1 were both target genes of miR-206 and miR-132. The levels of BDNF (29.50 μg·L^-1± 3.13 μg·L^-1) and SIRT1 (1.86 μg·L^-1± 0.25 μg·L^-1) in serum of the patients in aMCI group were both obviously lower than those in control group (BDNF:32.29 μg·L^-1±3.66 μg·L^-1; SIRT1:2.10 μg·L^-1± 0.29 μg·L^-1, P<0.05). Conclusion: The expression levels of miR-206 and miR-132 in serum of the aMCI patients are significantly up-regulated. Both of them might be involved in the pathogenesis of aMCI through inhibiting the BDNF and SIRT1 expressions.

Objective: To investigate the wear resistance of four low-shrinkage commercial composite resins and two traditional composite resins in vitro and discuss the relative influencing factors, and to illustrate the inner relationship of the wear resistance of resins and material composition. Methods: Four low-shrinkage commercial composite resins including CLEARFIL MAJESTY Posterior(CMP), Filtek LS(LS), Admira(AD), Kalore(KA) and two traditional composite resins including Filtek Z350XT (Z350), Solitaire2(S2) were chosen. The bar-shaped specimens were fabricated and mounted in a UMT-2 wear testing machine and abraded with the two-body media (distilled water) with a Si3N4 ball as antagonist.The maximum wear depth was determined after 14 400 cycles. The friction coefficient was determined during the test. The worn surfaces were examined with SEM. Results: CMP showed the lowest maximum wear depth and KA presented the highest maximum wear depth. The maximum wear depth ranked as follows:CMP< S2≈LS< Z350≈AD< KA(P<0.05). The wearing surfaces of Z350, S2, AD and KA were flatter and smoother, showing attrition wear and adhesive wear, whereas the wearing surfaces of CMP and LS were rougher, showing fatigue wear. Conclusion: The wear resistance of composite resins correlated to their type of filler, filler content and filler-resin matrix bonding.

Objective: To analyze the influencing factors associated with the malignant thyroid neoplasms and the level Ⅵ lymph node metastasis of papillary thyroid cancer(PTC),and to clarify the risk factors of the malignant thyroid neoplasms and level Ⅵ lymph node metastasis of the patients with PTC. Methods: A retrospective analysis on the materials of 99 PTC patients with routine thyroid nodular thyroid cancer resection and level Ⅵ lymph node dissection and 22 nodular goiter patients with thyroid lobectomy were performed.The PTC patients were divided into lymph node metastasis group (48 cases) and non-lymph node metastasis group (51 cases) based on the pathologic results. The associations between the clinical factors such as gender, age, tumor size, calcification, border and malignant thyroid neoplasms and the level Ⅵ lymph node metastasis of PTC were analyzed using Pearson χ² test. Results: Gender (P=0.959, OR =0.972)had no significant correlation with malignant thyroid neoplasms,but age (P<0.05, OR= 0.341), calcification (P< 0.001, OR = 0.115) and unclear border (P< 0.001, OR = 18.947) were associated with malignant thyroid neoplasms. Gender(P=0.379, OR = 1.523), calcification(P=0.064, OR =2.649) and unclear border(P=0.536, OR =0.727)were not apparently correlated with level Ⅵ lymph node metastasis of PTC, but age (P< 0.01,OR = 3.498) and tumor size (P< 0.001, OR = 0.177) were associated with level Ⅵ lymph node metastasis in the PTC patients. Co-existence of calcification and unclear border(P=0.189, OR=1.781)had no correlation with level Ⅵ lymph node metastasis in the PTC patients. Conclusion: Age is not only a risk factor for malignant thyroid neoplasms, but also a factor associated with level Ⅵ lymph node metastasis in the PTC patients. Calcification and unclear border are also the risk factors for malignant thyroid neoplasms, but they can not determine the central compartment lymph node metastasis in the PTC patients. In addition, tumor size can be prompted to predict the level Ⅵ lymph node metastasis in the PTC patients.

Objective: To investigate the expression of fibrillin-1 (FBN-1) in the atrium tissue of the patients with rheumatic heart valve disease complicated with atrial fibrillation(AF), and to explore its relationship with atrial fibrosis in the patients with valvular atrial fibrillation. Methods: Eighty-four consecutive patients with rheumatic heart valve disease underwent cardiac surgery were enrolled in this study. The patients were divided into AF group(n=39) and sinus rhythm group(SR group, n=45). The clinical data of patients were collected before operation. The right atrium tissue (0.3-0.5 mm³) was disserted during operation. The degrees of right atrial fibrosis of the patients in two groups were observed by Masson staining. Western blotting method was used to measure the protein expressions of FBN-1 in atrium tissue of the patients in two groups. Results: There were no significant differences in the gender ratio, age, blood pressure, blood biochemical indicators and other aspects of medical history between two groups(P>0.05); the diameters of left and right atrium of the patients in AF group were significantly larger than those in SR group(P<0.05). The Masson staining results showed that there was obvious fibrosis in AF group, and the collagen volume fraction and collagen level in AF group were significantly higher than those in SR group (P<0.05). The expression level of FBN-1 in right atrium tissue in AF group was obviously higher than that in SR group(P<0.05). The expression level of FBN-1 protein in right atrium tissue of the patients with valvular atrial fibrillation was positively correlated with the collagen level(r=0.544,P=0.021). Conclusion: There is obvious atrium fibrosis in the patients with valvular atrial fibrillation; it is closely related to the up-regulation of the expression of FBN-1 gene.

Objective: To probe the therapeutic effects of Butylphthalide Injection in the elderly patients with acute cerebral infarction (ACI) and its influence in cerebral hemodynamics and cerebral vascular reserve (CVR),and to clarify the pharmacological action mechanism of butylphthalide in treatment of ACI. Methods: A total of 100 cases of elderly patients with ACI were selected as the subjects and divided into observation group and control group according to the serial number on admission.Fifty cases were included in each group. The patients in control group were treated with the conventional treatment, while the patients in observation group were treated with Butylphthalide Injection on the basis of the conventional treatment. The National Institute of Health Stroke Scale (NIHSS) score, the brain hemodynamics indexes of the peak velocity (Vp), the mean velocity (Vm) and the differences of the velocity (DVp, DVm) as well as pulsatility index (PI), CVR of bilateral middle cerebral artery (MCA)of the patients in two groups were observed and compared.The therapeutic effects of the patients in two groups were evaluated and compared. Results: The NIHSS score of the patients in observation group after treatment was significantly lower than that in control group (t=15.420, P<0.05). The therapeutic effects and the clinical efficiency of the patients in observation group were significantly better than those in control group (U=2.225, χ²=5.005, P<0.05).The Vp and Vm of the patients in observation group after treatment were significantly higher than those in control group(t=10.819,t=7.259, P<0.05) and the DVp and DVm were significantly lower than those in control group (t=16.438,t=19.055, P<0.05). The CVR of the patients in observation group after treatment was significantly higher than that in control group(t=6.884, P<0.05)and the PI was significantly lower than that in control group (t=4.979, P<0.05). Conclusion: Butylphthalide Injection can effectively correct the abnormality of brain hemodynamics in the ACI patients, enhance the ability of body in maintaining the stability of cerebral vascular perfusion, improve the neurological symptoms in the patients with ACI, and improve the therapeutic effects.

Objective: To detect the interleukin-22(IL-22) levels in peripheral blood of the asthma patients and analyze the relationship between the IL-22 levels and IgE,pulmonary function,fractional exhaled nitric oxide(FENO),and to investigate the role of IL-22 in the pathogenesis of bronchial asthma. Methods: Seventy-eight patients with asthma were selected as experiment group. The asthma patients were divided into increased IgE group (n=46)and normal IgE group(n=32)according to the IgE levels.The asthma patients were also divided into acute attack stage and clinical control stage according to patients' clinical symptoms.In addition, twenty healthy controls were chosen as healthy control group. The plasma IL-22 levels, IgE,forced expiratory volume in one second (FEV1) and FENO levels of the subjects in various groups were detected and compared between various groups. The correlations between the plasma IL-22 levels and IgE, FEV1,FENO were analyzed. Results: The plasma IL-22 level and FENO level of the patients in asthma group were higher than those in healthy control group(P<0.05).The IL-22 level and FENO level of the patients in clinical control stage were lower than those in acute attack stage,but they were still higher than those in healthy control group(P<0.05).The plasma IL-22 level of the patients in increased IgE group was higher than that in normal IgE group(P<0.05). There were significant negative correlations between the IL-22 levels and FEV1 in the asthma patients in acute attack group and clinical control group(r=-0.365 3,P<0.05;r=-0.267 2,P<0.05),but there was no significant correlation between the IL-22 levels and FENO(r=0.011 9,P>0.05;r=0.059 8,P>0.05). Conclusion: The increased expression of IL-22 in peripheral blood of the patients with asthma may participate in the pathogenesis of asthma,and may aggravate the deterioration of pulmonary function of the asthma patients.

Objective: To detect the expressions of p16INK4a protein in the cervical lesion tissues of the Mongolian patients, and to explore the relationship between its expression and the occurrence and development of cervical cancer in the Mongolian patients. Methods: A total of 100 cases of paraffin sections of cervical cancer, cervical intraepithelial neoplasia(CIN),chronic cervicitis and uterine leiomyoma were divided into 25 cases of cervical cancer, 35 cases of CIN, 20 cases of chronic cervicitis, and 20 cases of uterine leiomyoma groups. The expressions of p16INK4a protein in different cervical tissues were detected by immunohistochemical SP method. Results: The positive rates of p16INK4a protern in cervical cancer, CIN, chronic cervicitis and uterine leiomyoma tissnes were 100.0%, 74.3%, 25.0%,and 10.0%, respectively.The results of K-WH rank sum test for multiple sample comparisons showed that the positive expression rate of p16INK4a protein in cervical cancer tissue was significantly higher than those in CIN, chronic cervicitis and uterine leiomyoma tissues(P<0.05). Conclusion: p16INK4a protein can be used as a indicator to screen the Mongolian patients with early cervical cancer.

Objective: To observe the influence of smoking in the levels of blood lipids and disease activity in the male patients with rheumatoid arthritis(RA), and to provide economical and effective intervention for the RA patients. Methods: A total of 112 male patients with RA were selected.The fasting blood samples from the ulnar vein of all patients were obtained in the morning. The level of C-reaction protein (CRP), erythrocyte sedimentation rate (ESR), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and other indicators were measured. The smoking status (including years of smoking, number of cigarettes smoked per day), tender joint count (TJC), swollen joint count (SJC) and other clinical data were collected and 28-joint disease activity score (DAS28) was calculated. The patients were divided into smoking group (87 cases) and non-smoking group (25 cases). According to the smoking index (SI)(years of smoking multiplied by number of cigarettes smoked per day), the patients were divided into group A (less than 400) (34 cases), group B (between 400 to 800) (37 cases) and group C (more than 800) (16 cases). Results: The levels of TC and TG of the patients in smoking group were significantly higher than those in non-smoking group(P<0.01), the HDL-C level was lower(P<0.01), and there was no significant difference in the LDL-C level between two groups(P>0.05). With the increase of SI, the levels of TC and TG were increased(P<0.05), and the HDL-C levels were decreased(P<0.05);the LDL-C levels were increased, but there was no significant difference between various groups(P>0.05). The levels of CRP and ESR in smoking group were higher than those in non-smoking group (P<0.05). The TJC, SJC, and DAS28 of the patients in smoking group were significantly higher than those in non-smoking group(P<0.01). With the increase of SI, the TJC, SJC, and DAS28 were increased(P<0.05), and the CRP and ESR levels showed an increasing trend. Conclusion: Smoking may cause the abnormal blood lipid levels in the patients with RA and the increased disease activity. To quit or reduce smoking may improve blood lipid metabolism in the patients with RA and has a positive effect on reducing the risk of RA.

Objective: To observe the influence of methylprednisolone in the respiratory mechanical parameters in the elderly patients undergoing major surgeries with general anesthesia intubation after mechanical ventilation,and to investigate whether methylprednisolone can improve the respiratory system degenerative diseases and benefit the mechanical ventilation. Methods: Sixty patients undergoing elective line laparotomy were divided into experimental group and conrol group (n=30) according to their wishes. The patients in experimental group receieved intravenous injection of methylprednisolone 1 mL (40 mg) after endotracheal intubation,and the patients in control group receieved 1 mL intravenous saline water injection.The airway peak pressure (Ppeak), airway platform (Pplat), lung compliance (Compl), and airway resistance (Raw) of the patients in two groups were recorded at the time of immediately before administration and 10,20,30,40 min after administration. The arterial blood oxygen partial pressure(PaO₂),arterial blood CO₂ partial pressure(PaCO₂),CO₂ partial pressure at the end of breathe out(PetCO₂) of the patients were detected at the time of immediately before administration and 20 min after administration. Results: Compared with before administration, the respiratory mechanics parameters of the patients in control group 10,20,30,40 min after administration had no statistically significant differences (P>0.05); the Comple of the patients in experimental group was singnificantly increased (P<0.05), and the Ppeak, Pplat,and Raw were significantly decreased (P<0.05).Compared with control group, the Compl of the patients in experiment group 10,20,30,and 40 min after administration were significantly increased (P<0.05),and the Ppeak, Pplat, and Raw were decreased (P<0.05). Compared with before administration,the PaO₂,PaCO₂,and PetCO₂ of the patients in control group 20 min after administration had no significant differences(P>0.05);the PaCO₂ and PetCO₂ of the patients in experimental group 20 min after administration were decreased(P<0.05),and the PaO₂ was increased(P<0.05).Compared with control group,the PaCO₂ and PetCO₂ of the patients in experimental group 20 min after administration were decreased(P<0.05),and the PaO₂ was increased(P<0.05). Conclusion: Methylprednisolone can obviously reduce PaCO₂,Peak, and Pplat and increase PaO₂ and Compl; methylprednisolone has possibility in improving the adverse effects of degenerative disease of respiratory system on general anesthesia in the elderly patients.

Objective: To explore the dose distribution characteristics between RapidArc and five-field intensity modulated radiotherapy(5F-IMRT) plans in the treatment of locally advanced pancreatic carcinoma,and to provide reference for selecting the appropriate radiation technique in clinic. Methods: Ten patients with locally advanced pancreatic carcinoma were selected.The patients were scanned by simulation CT, and the targets and organs at risk were contoured. RapidArc plan and 5F-IMRT plan were designed respectively. The treatment time and the differences of dose distribution in the targets and organs at risk of RapidArc and 5F-IMRT plans were compared. Results: The conformal index (CI) of RapidArc plan was superior than that of 5F-IMRT plan(P=0.01).The homogeneity index (HI) of targets were similar between two plans (P>0.05). RapidArc plan decreased the maximum dose (D_max) of the spinal cord(P=0.005); RapidArc plan decreased the mean dose(D_mean) of stomach(P=0.019); 5F-IMRT plan decreased the V20 of kidney(P=0.043);RapidArc plan decreased the mean dose(D_mean) of small intestine(P=0.011).The small intestine V10 and V20 of RapidArc plan were lower than those of 5F-IMRT plan(P=0.015,P=0.14); the monitor unit (MU) of RapidArc plan was significantly lower than that of 5F-IMRT plan,with a 18% reduction from the MU level of the latter one,and the treatment time was reduced by 70.3%. RapidArc plan had the smaller doses at liver and kidney compared with 5F-IMRT plan. Conclusion: For the patients with locally advanced pancreatic carcinoma,RapidArc plan has higher CI. RapidArc plan shows the advantages in the protection of organs. Compared with 5F-IMRT,RapidArc plan has less treatment time and significantly improves the curative efficiency.In clinic, the RapidArc plan is recommended.

Objective: To analyze the clinical characteristics of the patients with hyperthyroidism complicated with acute cerebral infarction, and to evaluate the prognosis and safety of intravenous thrombolysis treatment in the patients with hyperthyrodism complicated with acute cerebral infarctron within time window for thrombolysis. Methods: The clinical materials of patients with acute cerebral infarction treated with intravenous tissue plasminogen activator (rt-pA)were retrospectively analyzed.All patients were divided into hyperthyroidism group (n=41) and non-hyperthyroidismin group (n=160) according to whether complicated with hyperthyroidism.The general imformation,the area of cerebral infarction and intracranial hemorrhage of the patients in two groups were analyzed. Results: The NHISS score of the patients in hyperthyroidism group was higher than that in non-hyperthyroidism group for the first time(P<0.05 ).The NHISS scores 7 d after treatment and the 90 d modified Rankin scale (mRS) scores had no statistical differences between two groups (P>0.05).The proportions of the different infarction sizes had significant differences between two groups (P<0.05).The patients with large area cerebral infarction in hyperthyroidism group accounted for 51.2%, and it accounted for 27.5% in non-hyperthyroidism group.The incidence of asymptomatic intracranial hemorrhage of the patients in hyperthyroidism group was higher than that in non-hyperthyroidismin group (P< 0.05),but there was no statistical difference in the incidence of intracerebral hemorrhage (ICH) and subarachnoid hemorrhage (SHA) between two groups (P > 0.05). Conclusion: The patients with acute cerebral infarction complicated with hyperthyroidism could benefit from intravenous thrombolysis, without the increasing of incidence of spontaneous intracranial hemorrhage (sICH ) and SHA.

Objective: To study the changes of oral health indexes in adolescent orthodontic patients under different brushing time, and to provide basis for the orthodontists to give effective dental health promotion in clinic. Methods: A total of 123 adolescent orthodontic patients were included, and they were divided into 1-3min, 3-5 min, 5-7 min and 7-10 min groups based on the brushing time recorded in the period of orthodontic treatment. The relationship between different brushing time and oral health indexes was evaluated 1,3,and 6 months after experiment, such as sulcus bleeding index(SBI),plaque index(PLI), debris index-simplified(DI-S) and enamel demineralization index(EDI). Results: Compared with 1-3 min and 3-5 min groups, the SBI, PLI, and DI-S of the patients in 5-7 min group were significantly reduced(P<0.05). There was no significant differences of SBI, PLI, DI-S, and EDI between 5-7 min group and 7-10 min group (P>0.05). Conclusion: Brushing teeth more than 5 min can improve the oral hygiene significantly and the best time is 5-7 min;the orthodontists should require the patients to brush teeth at least for 5 min.

Objective: To discuss the clinical effects of implantation of radioative ¹²⁵I seeds by the way of percutaneous puncture and laparotomy under the guidance of ultrasound in the treatment of locally advanced pancreatic cancer, and to provide the basis for choosing surgical methods in treating advanced pancreatic cancer. Methods: The clinical materials of 73 patients with advanced pancreatic cancer were collected, including 42 patients who underwent implantation of radioactive ¹²⁵I seeds by percutaneous puncture(group A) and 31 patients who underwent impantation of radioactive ¹²⁵I seeds by laparotomy(group B). The pain relief, local control of tumor, postoperative survival time and complications of the patients were compared between two groups. Results: The rates of pain relief of the patients in group A and group B were 91.89% and 86.40%,and there was no significant difference(P=0.815).The local control rates of the patients in group A and group B were 71.43% and 77.42%,and there was no significant difference(P=0.564).The medium survival time of the patients in group A and group B were 11 months and 12 months; the one-year survival rates were 36.9% and 35.8%, and there was no significant difference(P=0.664). Seven patients in group A got fever; in group B, six patients got fever, two got calf muscle venous thrombosis, one got gastric retention, one got bilioentric anastomosis, one got abdominal distension and one got intestinal obstruction in the early stage after operation. The incidence rates of complications of the patients in two groups were 16.67% and 38.71%, and there was significant difference(P=0.034). Conclusion: Percutaneous implantation of radioactive ¹²⁵I particles guided by ultrasound causes less complications in the treatment of locally advanced pancreatic cancer. Moreover, the percutaneous way reaches the same effect as the intraoperative way does on the pain relief, local control of tumor and survival time prolonged.

Objective: To compare the efficacy and adverse reactions between intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid and simple intralesional betamethasone injection and topical hyaluronic acid in treatment of keloid. Methods: A total of 58 patients with keloid were divided into two groups:intralesional betamethasone injection combined with botulinum toxin type A and topical hyaluronic acid group (combined group, n=28) and intralesional betamethasone injection combined with topical hyaluronic acid group (control group, n=30). In both two groups, the intralesional betamethasone injection was given once every 4 weeks for 3 times totally, and topical hyaluronic acid was applied every day. In combined group, an injection of botulinum toxin type A was given around the lesion after the first intralesional injection of betamethasone. The parameters for therapeutic efficacy and adverse reactions of the patients in two groups were recorded, and the clinical pictures were taken before and after each treatment, and they were analyzed and compared. Results: After 3 treatments, compared with control group, the lesion appearance in combined group was improved. In control group, the VAS value was reduced at 1 month, but was increased gradually at 2 and 3 months. In combined group, the VAS value was reduced gradually in 3 months. There was a significant difference in VAS value at 2 and 3 months between two groups (P<0.05). In combined group, the thickness of keloids were reduced gradually in 3 months, while the reduction was not obvious in control group (P<0.05). The recurrence of pain, itching and lesion height were observed 2 weeks after each treatment in control group, but there was no recurrence in combined group. The incidence rate of adverse reactions was 26.7% in control group, which had no significant difference compared with combined group(25.0%) (P>0.05). Conclusion: Compared with control group, the combined group shows better efficiency in the treatment of keloid and there is no increase of adverse reactions. The therapy is worthy of clinical application.

Objective: To investigate the curative effect and safety of Stoppa approach combined with iliac fossa approach in the treatment of the patients with coxafossa anterior column or coxafossa dual column bone fracture, and to provide a scientific evidence for its application in the treatment of the patients with coxafossa anterior column or coxafossa dual column bone fracture. Methods: Sixty-three patients with coxafossa anterior column or coxafossa dual column bone fracture were selected; among them 32 cases were treated with stoppa approach combined iliac fossa approach(Stoppa group),and 31 patients were treated with ilioinguinal approach(control group).The intraoperative blood loss volume,the operaton time,the reset quality of fracture and hip joint function after operation of the patients in two groups were evaluated. Results: The intraoperative blood loss volume of the patients in Stoppa group was lower than that in control group, and the difference was statistically significant (P<0.05). The operation time of the patients in Stoppa group was shorter than that in control group, and the difference was statistically significant (P<0.05).The reset quality of fracture of the patients in stoppa group was higher than that in control group, and the difference was statistically significant (P<0.05).The hip joint function of the patients in Stoppa group was higher than that in control group, and the difference was statistically significant (P<0.05). Conclusion: Stoppa approach combined with iliac fossa approach is an effective method in the treatment of the patients with coxafossa anterior column or coxafossa dual column bone fracture with sufficient exposure and tiny tissue damage.Stoppa approach combined with iliac fossa approach in the treatment of the patients with coxafossa anterior column or coxafossa dual column bone fracture has the advantages of shorter operative time,less blood loss, higher rate of anatomic reduction,and good joint function.

Objective: To analyze the clinical effect and influencing factors of closed reduction and percutaneous cannulated screw fixation in the treatment of femoral neck fracture in the adolescents, and to provide evidence for surgical treatment of femoral neck fracture in the adolescents. Methods: The clinical effect and influencing factors of 36 cases of femoral neck fracture treated by closed reduction and percutaneous cannulated screw fixation were analyzed. The time of operation, the time of fracture healing evaluated with X-ray image, the evaluation on the function after operation by Harris score of hip joint,and the occurrence of complications of the patients were observed. Results: All the cases were followed up for 6-24 months, average 16 months. One case of all patients had avascular necrosis of the femoral head, and accounted for 2.78% of all the patients;2 cases of all fractures were nonunion, and 5.56%.The remaining 33 cases were completely healed.The HHS-harris hip score results showed that 26 were excellent, 6 cases were good, and 2 cases were poor;the excellent and good rate was 88.89%. Conclusion: Closed reduction and percutaneous cannulated screw fixation in the treatment of adolescent femoral neck fracture is an effective program of operation;timing of operation, good reduction, and rigid internal fixation can decrease the femoral head necrosis and the incidence of complications.

Objective: To collect the clinical materials of primary liver cancer patients treated by gamma knife and analyze the treatment methods as well as short-term efficacy,and to provide a reference for the clinical treatment of primary liver cancer patients. Methods: A total of 633 patients with inoperable primary liver cancer were treated by body gamma knife.According to the TNM staging method of Union for International Cancer Control(UICC), there were 351 cases with clear TNM staging.Among them, there were 251 cases (71.5%) at T3 stage and 57 cases(16.2%) at T4 stage.The prescription dose of 200-600 cGy each time to the 40%-85% dose line covering the planned target volume(PTV), this program was performed 5 times per week, and the number of treatment ranged from 2 to 13.The biochemical and imaging changes were observed 2-3 months after treatment to evaluate the short-term efficacy. Results: During the treatment, 229(36.2%)patients had adverse reactions,100 (15.8%) patients appeared the reduced white blood cells, and 137 (21.6%) patients appeared the reduced platelets.On discharge from the hospital, 601 patients were improved, 22 patients had no obvious change, 5 cases were worse, and 5 cases died.The proportion of improved patients who received the cumulative dose between 3 000 cGy to 4 000 cGy was higher than those who received the cumulative dose less than 3 000 cGy(P<0.05).After treatment, the curative effects were evaluated in 45 patients during two and three months, including PR 77.8%(35/45) and SD 22.2%(10/45), and the total effective rate was 77.8%(35/45).No statistical differences in effective rates were found between different tumor diameter, single dose, and cumulative dose groups(P>0.05). Conclusion: The proportion of adverse reactions in the primary liver cancer patients treated with body gamma knife is relatively low and the short-term efficacy is ideal.Body gamma knife treatment is a safe and effective treatment method for the primary liver cancer patients.

Objective: To investigate the application of lateral upper arm free flap in the repair of postoperative soft tissue defects after operation in the patients with tongue cancer, and to evaluate its survival rate and repair effect. Methods: Ten patients with tongue cancer, having a desire to repair soft tissue defect after tongue cancer operation, and their systemic state could tolerate the long time operation, were selected to repair tongue cancer postoperative defects with lateral upper arm flaps. Among them, 7 cases were male, 3 cases were female; aged 45-67 years old, average 52.1 years old.Radical resection of tongue cancer was primarily finished, then the flaps were designed and prepared to repair the defects of tongue according to the location and size of defects.After the operation, the survival rate of the flap was observed, and the effect of repair was evaluated according to the shape, size, volume and movement of the tongue. Results: The lateral upper arm flaps were stable and simple to prepare, the donor sites were concealed, and the upper limbs were not abnormal.All of flaps survived, and all of wounds healed primarily.The shape, size, and texture of tongues were satisfied, and the movements of tongue were not significantly restricted.After repair, the voice, mastication and swallowing function of the patients were good without obvious influence. Conclusion: The survival rate of lateral flap of upper arm is high, and the tongue shape and function were good after tongue defect repair.It is an ideal free flap for repairing the tongue tissue defect of tongue cancer after operation.

Objective: To report one case of mushroom poisoning complicated with multiple organ disfunction syndrome treated by sequential blood purification,and to explore the treatment method and principle of mushroom poisoning complicated with multiple organ disfunction syndrome. Methods: Hemoperfusion was performed in the patient,once every other day,combined with sequential blood purification of hemoperfusion and hemodialysis, and supplemented by hormone therapy and anti-infection. Results: The patient was out of danger,and the vital signs were stable;liver function,kidney function and blood routine were recovered. Conclusion: Early application of hemoperfusion combined with sequential blood purification can effectively treat the mushroom poisoning complicated with multiple organ disfunction syndrome and thrombotic microangiopathy,and improve the prognosis.

Objective: To observe the clinical effect of TOMY self-ligation appliance technique in the treatment of anterior crossbite and moderate crowding, and to explore its clinical application. Methods: A patient, manifested as anterior crossbite and moderate dentition crowing, diagnosed as Angle class Ⅲ subclass malocclusion, MaoⅡ1+Ⅰ1 malocclusion, high angle,and facial asymmetry, was selected. This patient was treated with TOMY self-ligation appliance technique without tooth extraction. The length and width of the dental arch, the width of the alveolar bone and the width of the basal arch were measured before and after treatment; the lateral radiographs of the patient were taken too. Results: After treatment,the upper and lower dentition arranged in neat rows, reached a neutral occlusal relationship;SNA, SNB and ANB had no obvious changes; the lip inclination of anterior teeth was increased by 10 degrees, and reached the normal value;the upper and lower dental arch width, alveolar bone width, basal bone arch width and dental arch length were lower than before treatment. Conclusion: For the patient with anterior crossbite and moderate dental crowding treated with non-extraction, the TOMY self-ligation technique can be considered to use, by increasing the arch width and length to improve the anterior crossbite and moderate crowding, in order to achieve the effective treatment.

Objective: To explore the etiology and treatment of one case of bilateral mandibular second molar impaction with paradental cyst, and to provide a reference for its diagnosis and treatment. Methods: Root canal treatment of the left mandibular first molar of the patient was performed before operation.The left mandibular second molar of the patient was removed; the residual dental follicle, the granulation tissue and the cyst wall were stroken off under local anesthesia.The diamond ball was used to polish the wound cavity and sharp bone edge, and to mill the distal apical part of left mandibular first molar.The tissue removed during the procedure was used for the pathological examination. Results: The X-ray image showed that the bilateral mandibular second molar was impacted with the left mandibular first molar root's absorption, and there was a clear round-like density reduction zone around the second molar crown.The pathologic result was paradental cyst. Conclusion: Dental impaction complicated with paradental cyst could occur in other tooth position except for the third molar.Its diagnosis should be combined with the clinical manifestations, the pathologic manifestations and the medical imaging.Multidisciplinary consultation is in favor of its diagnosis and treatment.

Objective: To investigate the clinical features of placental thrombosis complicated with fetal growth restriction(FGR),and to analyze its diagnosis and treatment methods. Methods: Combined with reviewing the relevant literatures, the clinical data of a case of placental thrombosis complicated with FGR was retrospectively analyzed. The patient with 32 1/7 weeks of gestation was hospitalized due to placental blood sinus found one month ago; at the same time FGR was found by ultrasound examination. The patient was intravenously given nutritional support treatment such as amino acid and glucose.At the same time, the patient was continuously given low-flow oxygen. Results: The patient received cesarean section at 35 2/7 weeks of gestation and a baby girl with 1 280 g weight and 32 cm length was gained;many blood sinus in the maternal surface of placenta were seen with the largest diameter of 3-4cm; the placenta was hypertrophic, weighted 540 g.After operation,the newborn was transferred to Department of Neonatology and followed up for 1 month.1 month later, the infant could eat by herself, other physical examinations were finished without any obvious abnormal findings and the newbron discharged from hospital after recovery. Conclusion: Placental thrombosis complicated with FGR is very common in clinic and this disease severely endangers the neonatal health. Early diagnosis and reasonable treatment can improve the pregnancy outcomes.

Objective: To evaluate the left ventricular segmental strains,global strains and systolic function in the patients with acute myocardial infarction (AMI) after percutaneous coronary intervention(PCI)with three-dimensional speckle tracking echocardiography(3D-STI),and to clarify the characteristics of early myocardial strain changes after PCI and their clinical values. Methods: Thirty patients received PCI for left anterior descending branch disease were enrolled as case group; at the same period, 30 healthy middleaged and elderly people were selected as control group.After two-dimensional ultrasound data were obtained regularly, the myocardial motion and systolic functions of the patients in two groups were detected with 3D-STI, and all segmental strain, global longitudinal strain(GLS), global radial strain(GRS), global circumferential strain (GCS)and global area strain(GAS) values of left ventricle and left ventricular ejection fraction (LVEF) values were collected. Results: Compared with control group, the global strain values of the patients in case group were lower than those in control group; 9/16 longitudimal strain(LS), 8/16 circumferential strain(CS), 5/16 radial strain(RS) and 11/16 area strain(AS) were significantly lower than those in control group(P<0.05).The decrease of strain value was mainly concentrated in the anterior segment of the anterior wall, the anterior segment of the anterior wall, anterior apical segment, anterior septal basal segment, anterior septal segment, posterior septal basal segment, posterior septal segment, and posterior septal segment.The GLS,GAS and GCS in case group were negatively correlated with LVEF(r=-0.819,r=-0.897,r=-0.807,P<0.0001);the GRS was postively correlated with LVEF(r=0.862,P<0.0001). Conclusion: 3D-STI can quantitatively analyze the segmental wall motion of left ventricle and could apply the parameters to evaluate the therapeutic efficacy after PCI.

Objective: To evaluate the reliability and validity of the adaptation OSSIQ in measuring the life quality of the populations with different types of subjective skin. Methods: A stratified samplings strategy was adopted to select 140 college students, workers, personnel of enterprises and institutions aged 18-60 years old.The reliability and validity of adaptation OSSIQ were evealuated using internal consistency validity, split half reliability, and construct validity. Results: The adaptation OSSIQ had a Cronbach's α efficient of 0.924 and 0.902. Split half reliability, the Spearman-Brown before and after skin care were 0.877 and the Guttman Split-Half coefficients were 0.876 and 0.874 respectively.Contentvalidity, the Item-scale correlation coefficients exceed 0.5 (P<0.001) before and after skin care.Satisfactory (100%) convergent validity and discriminant validity were found.The contract validity was checked by factor analysis; after quartimax rotation, the researcher extracted the factors by the default principle of eigenvalue>1.Each factor consisted of the items with factor loading≥0.6;the load factor value belonged to high level (≥0.6) by factor analysis. Conclusion: Adaptation OSSIQ has good reliability and validity for the evaluation on life quality of the populations with different types of subjective skin.It can be used to evaluate the influence of different types of skin in the life quality.

Objective: To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform/PEG method, and to provide basis for obtaining the batch of IgY. Methods: The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed. Results: The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity. Conclusion: The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.