Objective: To study the lethal effect of vitamin C carbon dots on oral squamous cell carcinoma KB cells, and to clarity its related mechanism.Methods: The KB cells were treated with different concentrations (5, 10, 20, 40 and 80 mg·L^-1) of vitamin C carbon dots in vitro as experimental groups, and 0 mg·L^-1 vitamin C carbon dots group was used as blank control group. MTT assay was used to detect the proliferation rates of KB cells in various groups; colony formation assay was used to detect the colony formation ability of KB cells; Western blotting was performed to detect the protein expression levels of autophagy related protein LC3 in KB cells in various groups; flow cytometry was used to detect the apoptotic rates of KB cells in various groups.Results: Compared with blank control group, the proliferation rates and colony formation abilities of KB cells in 20, 40 and 80 mg·L^-1 carbon dots groups were markedly decreased (P<0.01). Compared with blank control group, the protein expression level of LC3 Ⅱ in 40 mg·L^-1 carbon dots group was increased(P<0.05); the apoptotic rate of KB cells was markedly increased(P<0.01).Conclusion: Vitamin C carbon dots can kill the oral squamous cell carcinoma KB cells effectively, suppress the proliferation and impair the colony formation ability of KB cells, which is related to autophagy and apoptosis of KB cells.

Objective: To explore the effects of exosomes derived from bone-marrow endothelial progenitor cells(EPCs-Exos)on the angiogenesis and collagen deposition in vitro,and to illustrate the possible mechanism for EPCs-Exostoaccelerate the cutaneous deficiency repair.Methods: The endothelial progenitor cells (EPCs) were isolated, cultivated and identified; at the same time, the EPCs-Exos were also isolated and identified. The EPCs-Exos uptake in EPCswas observed and analyzed.16 rats were randomly divided into 4 groups, and then the models of skin defect were established, respectively. The equal volume of PBS and 50, 100, 150 mg·L^-1 EPCs-Exos were injected into the area around skin defect of the rats in 4 groups. The wound closed sizes on the 0, 3rd, 7th and 14th days were measured, and Masson's trichrome staining and CD31 immunofluorescence staining were performed on the 14th day for evaluating the tissue healing efficacy after EPCs-Exos treatment. In vitro,the mediums containing PBS and 50, 100, 150 mg·L^-1 EPCs-Exos were used to culture the human umbilical vein endothelial cells (HUVECs), respectively. Scratch test and tubule formation assay were used to detect the migration and capillary network formation of HUVECs. At the same time, Western blotting was used for analyzing the expression level of angiogenesis related gene vascular endothelial growth factor A (VEGFA) in HUVECs.Results: The primary EPCs were isolated and identified successfully, and EPCs-Exos were purified and characterized.The CD31 immunofluorescence staining and double staining of DiL-ac-LDL and FITC-UEA-I of EPCs were positive. The electron microscope results showed that EPCs-Exos were nearly spheroidal, with the diameter about 40-100 nm. For the models of rat skin injury treated by EPCs-Exos, with the increasing of injection doses, the sizes of skin defect scar were gradually reduced, the degrees of scar healings were gradually increased,and the differences between various groups were statistically significant (P< 0.05). EPCs-Exos promoted the collagen maturity of healing skin in a dose-dependent manner; on the 14th day, the effect in 150 mg·L^-1 EPCs-Exos group was the most significant. In vitro, EPCs-Exos promoted the migration and capillary network formation of HUVECs and increased the expression level of VEGFA; the migration rate,the net number of branches and the expression level of VEGFA in 150 mg·L^-1 EPCs-Exos group were significantly higher than those in 50 mg·L^-1 EPCs-Exos group and PBS group (P< 0.05).Conclusion: EPCs-Exos can promote the repair of traumatic skin defect of the rats by positively regulating the vascular endothelial cell function.

Objective: To modify Ganodermalucidum polysaccharides(GLP) with sulfate and observe the protective effect of Ganodermalucidum polysaccharide sulfate (GLPS) on the cerebral ischemia reperfusion injury in the rats,and to investigate its mechanism.Methods: GLP was modified by sulfation to obtain GLPS. A total of 100 SD rats were randomly divided into sham operation group, model group, GLP group (40mg·kg^-1·d^-1), GLPS group (40mg·kg^-1·d^-1) and nimodipine group (1mg·kg^-1·d^-1).The cerebral ischemia reperfusion models were established by middle cerebral artery occlusion method in the rats. The neurologic deficit score and the content of water in brain tissue of the rats with cerebral ischemia reperfusion injury were detected and the activities of superoxide dismutase(SOD) and the levels of malondialdehyde (MDA) were detected. The levels of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB),tumor necrosis factor-alpha (TNF-α), interleukin-1 (IL-1) and interleukin-6 (IL-6) in the brain tissue homogenate were detected by ELISA. Western blotting method was used to detect the protein expression levels of HSP70 and p-Akt in the brain tissue of the rats.Results: Compared with model group, the neurological function scores of the rats in GLP group and GLPS group were decreased(P<0.01),the water contents in brain tissue were decreased(P<0.05), the SOD activities were increased and the MDA levels were decreased(P<0.05), and the levels of NF-κB, TNF-α, IL-1 and IL-6 were decreased(P<0.05);the effect in GLPS group was significantly better than that in GLP group(P<0.05). The results of Western blotting method showed that the p-Akt protein expression levels in the brain tissue of the rats in GLP and GLPS groups were increased compared with model group (P<0.05); compared with model group, the HSP-70 protein expression level in the brain tissue of the rats in GLPS group was increased(P<0.01),but the effect in GLP group was not obvious.Conclusion: Sulfation can significantly improve the protective effect of GLP on the cerebral ischemia reperfusion injury in the rats and its mechanism may be related to regulating the HSP70/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.

Objective: To observe the effects of myricetin on autophagy and mitochondrial fission in the human ovarian cancer SKOV3 cells,and to explore its induction effect on autophagy and promoting effect on mitochondrial fission.Methods: The SKOV3 cells were cultured in vitro.0,20,40,and 60 g·L^-1 myricetin were used in control group and low, middle, high doses of myricetin groupsfor 12 h. The changes of mitochondrial membrane potential were detected by flow cytometry; the morphology of mitochondria was observed by MitoTracker Red; the expression levels of dynamin related protein1 (Drp1) and fission 1 (Fis1) in various groups were detected by Western blotting method; and the expression levels of autophagy related protein LC3 were detected by both Western blotting method and immunofluorescence method.Results: Compared with control group, the ratios of decreased mitochondria in different doses of myricetin groups were significantly increased(t=3.27, t=6.85, t=5.49,P<0.05).Compared with control group, the numbers of mitochondria in different doses of myricetin groups were increased, and the mitochondria looked more like gravel.Compared with control group,the expression levels of Drp1 in different doses of myricetin groups were significantly increased (t=4.35, t=3.28, t=6.17,P<0.05), and the expression levels of Fis1 were increased also(t=8.32, t=6.74, t=9.27).The immunofluorescence results showed that the expression levels of LC3 in different doses of myricetin groups were significantly increased with the increase of myricetin dose compared with control group. The Western blotting results showed that the ratios of LC3-Ⅱ/LC3-Ⅰ in middle and high doses of myricetin groups were significantly increased compared with control group(t=3.28, t=4.21,P<0.05).Conclusion: Myricetin can induce the autophagy of SKOV3 cells, and it can also promote the mitochondrial fission.

Objective: Toscreenthe proteins interacting with the human cytomegalovirus(HCMV)UL132 protein from the human fetus brain cDNA library by using Yeast Two-Hybrid System, and to elucidate the possible mechanism of UL132 protein in congenital cytomegalovirus infection.Methods: The HCMV UL132 fragment was amplified by polymerase chain reaction,the amplified HCMV UL132 fragment and expression vector pGBKT7 were digested and purified,and the HCMV UL132 fragment was linked to the vector pGBKT7.The pGBKT7-UL132 was constructed and transformed to yeast AH109, then the Human Fetal Brain DNA Library DNA was transformed into AH109 yeast. Using HCMV UL132 as abait, a human fetus brain cDNA was screened and the proteins interacting with UL132 protein were searched, the positive clone was sequenced and analyzed by bioinformatics methods.Results: The bait expression vector pGBKT7-UL132 was successfully constructed. The results of double enzyme digestion showed that there were two visible bands of 800 and 7 000 bp, respectively.After transformation of library plasmid, the transformation efficiency was calculated, and the transformation efficiency was 6.6×10³ cfu·μg^-1. There were 95 blue clones by X -gal coloration reactionsequencing and there were 10 clones interacting with the protein encoded by UL141 protein.The BLAST analysis showed that 7 of them were highly homologous with CAML.Conclusion: CAML might be one interaction protein with HCMV UL132 in Human Fetus Brain cDNA Library,suggesting that the interaction may be associated with the invasion and proliferation of the HCMV.

Objective: To construct lentiviral vector which can overexpression miR-137and produce lentivirus by lentivirus packaging system, and to explore its infection efficiency and expression in HEK293T cells.Methods: miR-137 sequence was chemically synthesized and cloned into lentiviral vector GV209, and the recombinant plasmid containing human miR-137 was obtained and identified. Then miR-137 recombinant plasmid together with two helper plasmids were transfected into HEK293T cells using Lipofectamine 2000.After the HEK293T cells were infected in multiplicity of infection(MOI) 40 for 48 h, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the expression level of miR-137 was detected by fluorescence quantitative PCR.Results: The sequencing results showed that the inserted gene sequence was completely consistent with the published human miR-137 gene sequence in GenBank. The GFP was observed in the HEK293T cells infected with miR-137 overexpression lentivirus under fluorescence microscope. The fluorescence quantitative PCR results showed that the expression level of miR-137 in the cells infected with overexpression lentivirus was 12.74 times higher than that in the control cells.Conclusion: The lentivirus containing miR-137 gene is successful packaged, and it could efficiently infect the HEK293T cells.

Objective: To determine the changes of the main components after compatibility of compound Mailuqishen(MLQS), and to explore its anti-tumor effect in the H22 tumor-bearing mice.Methods: The contents of ginsenoside and amino acid in MLQS were detected by HPLC, and the content of polysaccharide was detected by phenol-sulfuric acid. A total of 144 female Kunming mice were randomly divided into normal control group, model group, positive control group, low dose of MLQS group, medium dose of MLQS group, high dose of MLQS group, ginseng geng group, ginseng group and antler plate group (n=16). Except normal control group, the mice in the other eight groups were used to establish the H22 tumor-bearing mouse models, then the mice were treated with drugs at next day. The tumor weights, inhibitory rates of tumor, spleen and thymus indexes of the H22 tumor-bearing mice were detected 10 d after administration. The morphological changes of tumor and spleen tissue were examined by HE staining, and the apoptotic rates of H22 tumor cells were tested by flow cytometry.Results: As calculation with the ginseng and antler plate single herb, the contents of ginsenosides, polysaccharides and amino acids in MLQS were significantly higher than those of single herbs (P<0.05). Compared with model group, the inhibitory rates of tumor in various administration groups were significantly increased(P<0.01); the spleen indexes and thymus indexes of the mice in different doses of MLQS groups were significantly increased(P<0.01); the apoptotic rates of tumor cells were markedly increased(P<0.05). Compared with model group, the tumor tissue of the mice in various administration groups was destroyed, the cells were sparse and irregular, and the tumor presented necrotic lesions; the morphology of spleen tissue was normal with discernible fringe, and the lymphocytes arranged densely.Conclusion: The contents of ginsenosides, polysaccharides, and amino acids in compound MLQS are significantly increased compared with those of single herbs, and its anti-tumor effect is stronger than the single herbs.

Objective: To detect the expressions of urea transporter B (UT-B) in tumor tissue of bladder cancer patients and normal urothelium tissue, and to investigate its clinical significance of expression in bladder cancer tissue.Methods: Fifty-two paraffin-embedded specimens of bladder cancer patients and 15 normal urothelium specimens were selected. The expression levels of UT-B protein were detected by immunohistochemistry method.The difference of expression of UT-B in bladder cancer tissue and normal urothelium tissue and its relationship with the clinicopathological parameters were analyzed.Results: The expression rate of UT-B protein in bladder cancer tissue was 44.2%, while it was significantly higher in control group (93.3%), and the difference between two groups was significant (P=0.001). The positive expression rates of UT-B in bladder cancer tissue were significantly decreased with the increasing of histological grades, and there were significant differences between different grade groups (P=0.010). The positive expression rate of UT-B in muscle-invasive stage (Ta+T1) was significantly lower than that in non-muscle-invasive stage (T2+T3) (P=0.014).Conclusion: The reduction or lack of UT-B expression may promote the incidence, progression and invasiveness of bladder cancer.

Objective: To study the expression levels of forkhead transcription factor(FoxO1) mRNA and protein in the insulin resistance (IR) HepG2 cells model (HepG2/IR) and IR reversal HepG2 cells model (HepG2/IR-PH), and to explore its mechanism in IR.Methods: The HepG2/IR was induced with different doses of insulin (1×10^-10, 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6 and 1×10^-5 mol·L^-1) for different time(24, 36 and 48 h)in the HepG2 cells.The cells in control group were not treated with insulin. The glucose levels in supernant were determined by glucose oxidase method, and the glucose consumption in HepG2 in various groups were calculated to confirm the optimum induction conditions of HepG2/IR. The HepG2/IR-PH was induced with different doses of pioglitazone hydrochloride (PH) (0.156, 0.313, 0.625, 1.250, 2.500, 5.000, 10.000 and 20.000 mmol·L^-1) in the HepG2 cells, and control group was set up at the same time. The proliferation activities of cells were observed by MTT assay to confirm the optimum reversal concentration of PH. The FoxO1 mRNA and protein expression levels were detected by Real-time PCR and Western blotting methods.Results: The glucose consumption decreased by 45.84% in HepG2/IR after treated with 1×10^-7 mol·L^-1 insulin for 36 h, and there was significant difference compared with control group(P<0.05), the proliferation activity of cells had no change in the HepG2/IR-PH after treated with 1.25 mmol·L^-1 PH for 24 h(P>0.05).Compared with control group, the expression levels of FoxO1 mRNA and protein in HepG2/IR were significantly increased(P<0.01); compared with control group, the expression levels of FoxO1 mRNA and protein in HepG2/IR-PH had no significant differences(P>0.05).Conclusion: The IR of HepG2/IR is associated with the FoxO1 mRNA expression. The detection of FoxO1 mRNA seems to be an indicator to evaluate the efficacy of insulin sensitizer, and inhibiting the expression of FoxO1 mRNA may be developed as a potential therapy for type 2 diabetes.

Objective: To transfect the tongue squamous cell carcinoma Tca8113 cells with NICD, and to investigate the effects of Notch 1 on Tca8113 cells and the expression of epidermal growth factor receptor (EGFR) in tongue squamous cell carcinoma.Methods: The Notch plasmid pRAMIC-IRES2-EGFP encoding exogenous and control plasmid pIRES2-EGFP were transiently transfected into the Tca8113 cells by Lipofectamine^TM 2000. In the experiment, target plasmid pRAMIC-IRES2-EGFP transfection group, control plasmid pIRES2-EGFP transfection group and non-transfection group were set up. RT-PCR, Western blotting and immunocytochemical method were used to observe the mRNA and protein expression levels of Notch 1 and EGFR. The proliferation activities of Tca8113 cells were detected by MTT assay,and the survival rates were calculated.Results: Compared with non-transfection group and control plasmid pIRES2-EGFP transfection group, the mRNA and protein expression levels of Notch 1 in Tca8113 cells in pRAMIC-IRES2-EGFP group was significantly increased(P<0.05), the mRNA and protein expression levels of EGFR in Tca8113 cells were significantly decreased (P<0.05),and the proliferation rate of Tca8113 cells was significantly decreased(P<0.05) after transfection. There were no significant differences in the expression levels of Notch 1 and EGFR mRNA and protein and the proliferation rates in Tca8113 cells between non-transfection group and pRAMIC-IRES2-EGFP group (P>0.05).Conclusion: The effect of Notch 1 on the proliferation of Tca8113 cells may be related to down-ragulatoin of expression level of EGFR.

Objective: To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods: HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus. The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method. The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group); the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×10⁹ pfu·mL^-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%. The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.

Objective: To observe the inhibitory effect of ischemic postconditioning (I-postC) on the apoptosis of renal cells after limb ischemia reperfusion(LIR) in the rats, and to investigate the possible mechanisms.Methods: Thirty SD rats were randomly divided control group, ischemia-reperfusion group(IR group) and I-postC group(n=10).4 h ischemia and 4 h reperfusion with the rubber band in two hind limbs of the rats were performed to establish the models. In control group, the rubber band around the limb was loose and the blood flow was not blocked. As for I-postC group, before perfusion, 5 min ischemia and 5 min reperfusion were performed in the rats and repeated 3 times named I-post C. The levels of blood creatinine (Cr), blood urea nitrogen(BUN) and C-reactive protein (CRP) in plasma of the rats in various groups were measured by automatic biochemistry analyzer. The expressions of Bcl-2 protein and Bax protein in renal were detected by immunohistochemical method,and its quantitative results were observed with automatic image analysis system and the ratio of Bcl-2/Bax was calculated.The apoptotic cells in kidney tissue were determined by terminal-deoxynucleotidy1 transferase-mediated d-UTP nick end labeling (TUNEL) under laser scanning confocal microscope(LSCM).The ultrastructures of kidney tissue were observed under electron microscope.Results: Compared with control group, the levels of Cr,BUN and CRP in plasma of the rats in IR group and I-postC group were increased(P<0.05 or P<0.01);compared with IR group, the levels of Cr, BUN and CRP in plasma of the rats in I-postC group were decreased(P<0.01).Compared with control group,the expression levels Bax and Bcl-2 in kidney tissue of the rats in IR group and I-postC group were significantly increased (P<0.05 or P<0.01),and the ratios of Bcl-2/Bax were reduced(P<0.05);compared with IR group,the expression level of Bax in kidney tissue of the rats in I-postC group was decreased (P<0.05),the expression level of Bcl-2 was increased(P<0.01),and the ratio of Bcl-2/Bax was increased(P<0.05).Under laser confocal microscope,the number of apoptotic cells in kidney tissue of the rats in IR group was increased compared with control group;the number of apoptotic cells in I-postC group was decreased compared with IR group.Under transmission electron microscope,the changes in IR group were found as follows:renal proximal convoluted tubule epithelial cell nucleus vacuoles,increased lysosome and dense particle deposition, some mitochondria crest fracture or fuzzy; irregular and fusion, glomerular podocyte protuberance,mitochondrial cristae fracture and reducetion with vacuoles, rough endoplasmic reticulum expansion; the damage levels of renal tubular epithelial cells and glomerulus in I-postC group were improved compared with IR group.Conclusion: Limb ischemia reperfusion can induce the apoptosis of renal cells, I-postC can inhibit the apoptosis of renal cells,and it would be helpful to improve the kidney function.

Objective: To study the influence of rutin in the morphology of renal tissue of the diabetic mice induced by streptozotocin(STZ), and to clarify the effect of rutin on the kidney tissue damage.Methods: Twelve mice of the total 70 Kunming mice were used as normal group, the other were used to estabish type 1 diabetes mouse models by intraperitoneally injected with STZ (62.5 mg·kg^-1), once daily for 5 d. The successfully established model mice were randomly divided into model group,low dose (50mg·kg^-1)of rutin group, high dose (100 mg·kg^-1) of rutin group and irbesartan group (45 mg·kg^-1). The mice in model group and normal group were given the carboxy methyl cellulose(CMC) and the other mice were given drugs by intragastric administration once daily for 8 weeks accordingly. The weight and blood glucose of the each mouse were determined.Full automatic biochemical analyzer was used to detect the levels of serum creatinine (Cre) and blood urine nitrogen (BUN) of the mice, and the kidney index was calculated. The morphology of renal tissue was observed by HE staining, Masson staining and electron microscope.Results: After injection of STZ,the model success rate was up to 98%.Compared with normal group, there was no significant difference in the weight of the mice in other groups before administration(P>0.05).After administration of rutin, the weights of the mice in model group, low dose of rutin group, high dose of rutin group and irbesartan group were significantly decreased (P<0.05). Compared with model group, the levels of blood glucose of the mice in low and high doses of rutin groups were significantly decreased(P<0.05);the levels of Cre and BUN were significantly reduced (P<0.05);the kidney index of the mice in high dose of rutin group was significantly decreased (P<0.05). Compared with normal group,the kidney tissue of the mice in model group was seriously damaged;glomerular was weaked, the kidney tissue fibrosis was serious, glomerular basement membrane was diffusely thickened and foot process was coalesced or overgrow. Compared with model group,the degree of injury of the mice in low and high doses of rutin groups were significantly improved, especially in high dose of rutin group.Conclusion: Rutin can improve the renal function of diabetic mice induced by STZ and reduce the degree of renal tissue damage in the diabetic mice

Objective: To establish rat diabetes and eye disease models by injection of STZ and explore the therapeutic effect of panaxnotoginseng polysaccharides (PNP) on the diabetes and eye diseases of the model rats,and to clarify their mechanisms.Methods: Seventy SD male rats were randomly divided into blank control (n=10) and model groups (n=60), and the rats in model group were fed with high fat diet. 2 weeks later, the rats in model group were intraperitoneally injected with 35 mg·kg^-1 STZ to establish the models. And 3 d later, the rats were treated with fasting and water deprivation for 12 h,the fasting blood glucose (FBG)was tested, and the models were assessed to be successful as the FBG>11.1 mmol·L^-1. The rats with hyperglycemia were selected and divided into model, melbine(150 mg·kg^-1), and low, middle and high doses (75,150 and 300 mg·kg^-1) of PNP groups. After orally administration for 5 and 8 weeks, the FBG levels of rats were recorded. And 8 weeks later, the sugar tolerance, hepatic glycogen levels,serum glutathione(GSH) and nitric oxide(NO) levels of the rats were tested. The rat retinas were removed to analyze the expression levels of vascular endothelial growth factor(VEGF) and inducible nitric oxide synthase(iNOS) by using Q-PCR. The pathological changes of retinas were observed by HE staining method.Results: Compared with model group,the FBG level in middle dose of PNP group was decreased 5 weeks after treatment(P<0.05). Eight weeks later, compared with model group, the levels of FBG, sugar tolerance and hepatic glycogen in different doses of PNP groups were all decreased (P<0.05 or P<0.01). Compared with model group, the level of serum GSH in high dose of PNP group was remarkably increased(P<0.01), and the NO level was significantly decreased (P<0.05 or P<0.01). Compared with model group, the expression levels of VEGF and iNOS in high dose of PNP group were significantly decreased (P<0.05). The results of HE staining showed that the neurodeatrophia of the rats in low and middle doses of PNP groups were improved; and the vascular proliferation and neurodeatrophia of the rats in high dose of PNP group were significantly improved.Conclusion: PNP could decrease the blood sugar, increase the levels of GSH and NO, and up-regulate the gene expression levels of VEGF and iNOS, resulting the treatment of diabetes and its related retinopathy.

Objective: To study the influence of gallic acid (GA) on the migration of human gastric carcinoma SGC-7901 cells, and to explore its mechanism.Methods: The human gastric carcinoma SGC-7901 cells were cultured and divided into control group and 3.125, 6.250, 12.500, 25.000, 50.000,100.000 mg·L^-1 GA groups. The inhibitory rates of proliferation of SGC-7901 cells in various groups were examined by MTT assay; the migration abilities of SGC-7901 cells in various groups were measured with scratch assay; the expression levels of vascular of endothelial growth factor (VEGF) in various groups were detected by immunocytochemistry.Results: Compared with control group, the inhibitory rates of proliferation of SGC-7901 cells in different doses of GA groups were significantly increased in a dose-dependent manner(F=59.451,P<0.01). Compared with control group, the wound healing rates in different doses of GA groups were significantly decreased (P<0.01). Compared with control group, the expression levels of VEGF protein in 12.500 and 25.000 mg·L^-1 GA groups were decreased (P<0.05).Conclusion: GA could inhibit the proliferation and migration of SGC-7901 cells through down-regulating the expression levels of VEGF protein.

Objective: To explore the changes of chemotherapy sensitivity of breast cancer MCF-7/ADM cells afterHiwi gene targeting silence, and to illuminate the role of Hiwi gene in resistant mechanism of breast cancer MCF-7/ADM cells.Methods: The MCF-7/ADM cells were transfected with Hiwi control,siRNA1 Hiwi gene and siRNA2 Hiwi gene as Hiwi control group, Hiwi siRNA1 group and Hiwi siRNA2 group.The expression levels of Hiwi gene mRNA and protein in breast cancer MCF-7/ADM cells were detected by Real-time PCR and Western blotting methods to judge the transfection rates in various groups.After transfection,the breast cancer MCF-7/ADM cells invarious groups were treated with the different concentrations(0,0.1,0.5 and 1.0 mg·L^-1) of adriamycin, and the cell resistant sensitivities were detected by MTT method.Results: Compared with control group(0 mg·L^-1 adriamycin), the expression levels of Hiwi gene mRNA and protein inMCF-7/ADM cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased (P<0.01).Compared with control group, the survival rates of Cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased after treated with different concentrations of adriamycin,and the survival rates in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased(P<0.01);they were (48.15±6.28)% and (41.73±6.17)% when the concentration of adriamycin was 1 mg·L^-1,and the sensitivities to the adriamycinwere obviously enhanced(P<0.01).Conclusion: The target silencing Hiwi genes in MCF-7/ADM cells is efficient.The sensitivity of MCF-7/ADM cells to adriamycin restores after Hiwi gene silencing.

Objective: To investigate the relationship between ciaH, eno, pykF genes and fluoride resistance through determining the differential expressions of ciaH, eno and pykF genes of fluoride-resistant Streptococcus mutans cultivated in fluoride environment.Methods: The cultured Streptococcus mutans and their fluoride-resistant strains were divided into UA (Streptococcus mutans subcultured in BHI without NaF), FR (fluoride-resistant Streptococcus mutans subcultured in BHI without NaF) and FFR (fluoride-resistant Streptococcus mutans subcultured in BHI containing 1 g·L^-1 NaF) groups. After 11 h (logarithmic phase) and 20 h (platform stage) cultivation, the expression levels of ciaH, eno and pykF mRNA were detected by RT-PCR method.Results: Compared with FR group, the expression levels of ciaH, eno and pykF mRNA in FFR group were increased both in the logarithmic phase and the platform stage(P<0.05 or P<0.01).Compared with UA group, the expression levels of eno and pykF mRNA in FR group were decreased both in the logarithmic phase and the platform stage(P<0.01), whereas the expression level of ciaH mRNA had no significant difference in the logarithmic phase (P>0.05), but it was increased in the platform stage (P<0.01).Conclusion: Fluoride can increase the expression levels of ciaH, eno and pykF genes in fluoride-resistant Streptococcus mutans, indicating that these genes are related to the production of fluoride resistance.

Objective: To investigate the changes of expression levels of TLR9 and IL-6 in the periodontal tissue during the experimental tooth movement of the rats, and the effecst of the MT01 on the expression of TLR9,TRAF6 and IL-6 in periodontal tissue,and to clarify its related mechanisms.Methods: Thirty Wistar rats were randomly divided into MT01 intervention group(n=6) and non-MT01 group(n=24). Force of 0.49 N was applied to move the upper first molars mesially. The rats in Non-MT01 intervention group were sacrificed on the days 3,7,14 and 21, and the rats in MT01 intervention group were all sacrificed on the day 7. RT-qPCR was used to detect the expression levels of TLR9,TRAF6 and IL-6 mRNA in maxillary first molar alveolar bone in each group.Results: The expression levels of IL-6 and TLR9 mRNA in loaded side were higher than those in control side(P<0.05), and reached the maximum levels on the day 7(P<0.01); with the interpose of MT01, the expression levels of TLR9 and TRAF6 mRNA were lower than control side(P<0.01).Conclusion: MT01 could down-regulate the expression levels of TLR9 and TRAF6 during orthodontic tooth movement and eventually resists the inflammation during the tooth movement.

Objective: To investigate the clinical and laboratory characteristics of lupus nephritis(LN) patients by detecting the anti-nucleosome antibodies, anti-C1q antibodies and anti-double stranded antibodies(anti-ds DNA), and to clarify the risk factors of LN in the patients with systemic lupus erythematosus(SLE),and the significance of three kinds of antibodies in diagnosis of LN.Methods: A total of 120 SLE patients were selected and divided into LN group(n=60) and non-LN group(n=60).The ANAS data of 120 patients were retrospectively analyzed,the levels of anti-C1q antibodies were measured.The clinical symptoms and laboratory data of the patients with positive anti-dsDNA, -nucleosome and -C1q antibodies (3-pos group)and negative three kinds of antibodies(non 3-pos group) were analyzed in LN group.Results: The positive rate of anti-C1q antibody of the patients in LN group (40.00%) was higher than that in non-LN group (21.67%) (χ²=4.728, P=0.03).The positive rate of anti-dsDNA antibody in LN group was 66.67%, and it was 46.67% in non-LN group; the positive rates of the patients had significant difference between two groups (χ²=4.887, P=0.027). The positive rate of anti-nucleosome antibody in LN group was 58.33%, and it was 40.00% in non-LN group; the positive rates of the patients had significant difference between two groups (χ²=4.034, P=0.045).The positive rates of U1-snRNP, SmD1 and other antibodies Jo-1, SSA/Ro60kD, SSA/Ro52kD, SSB, ScL-70, CENP-B,and P0 had no significant differences between two groups(P>0.05). The levels of C3 and C4 and hemoglobinin of the patients in 3-pos group were higher than those innon 3-pos group (P<0.05);the age,the levels of immunoglobulin protein and C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR), white blood cell (WBC) and platelet had no statistically significant differences between 3-pos and non 3-pos groups(P>0.05).The clinical symptoms were not statistically significant in 3-pos and non 3-pos groups (P>0.05).Conclusion: The anti-nucleosome, anti-C1q and anti-dsDNA antibodies are the risk factors of SLE complicated with LN;the positive antibodies can improve the diagnostic rate of LN.The 3-pos patients have more severe damage in complements and blood system with higher renal disease activities.

Objective: To systematically review the efficacy and safety of dexmedetomidine and midazolam in procedural sedation.Methods: PubMed,EMBase,Cochrane Library,CNKI,CBM and WanFang databases were retrieved to collect the randomized controlled trials (RCT) about comparion of efficacy and safety between dexmedetomidine and midazolam in procedural sedation up to March, 2017. Based on the inclusion criteria, the data extraction and quality evaluation were performed, and then the systematic evaluation was carried out.The outcome measures for efficacy were the satisfaction scores and pain scores of the patients and clinicians;the outcome measures for safety comparison were hypotension, hypoxia, and circulatory and respiratory complications.Results: There were 14 RCT satisfied the inclusion criteria including 949 patients. Compared with midazolam group, the incidence of pain, delirium, and analgesia of the patients in dexmedetomidine group had significant differences (P<0.05); but the incidence of respiratory depression, low blood pressure had no significant differences (P>0.05).Conclusion: When the adult patients are sedated, dexmedetomidine can be used as an ideal alternative to midazolam sedation.

Objective: To detect the expression and clinical significance of β-tubulin Ⅲ in cancer tissue of the patients with colon adenocarcinoma,and to explore its clinical significance.Methods: A total of 111 colon adenocarcinoma tissue samples were obtained. According to the location of β-tubulin Ⅲ positive cells, all patients were divided into front group (n=72) and non-front group (n=39). The positive expression rate of β-tubulin Ⅲ in the patients with colon adenocarcinoma was detected with immunohistochemistry. The correlations among the expression of β-tubulin Ⅲ and gender,age,tumor differentiation, clinical stage, lymph node metastasis, recurrence and death were analyzed.Results: The expression levels of β-tubulin Ⅲ had no significant differences between the patients with different gerder,age,lymph node metastasis,clinical stages,death and recurrence.The positive expression rates of β-tubulin in cancer tissue of the patients had significant difference between front and non-front groups (χ²=8.76, P=0.01). Lowly-to-moderately differentiated tissue was more common in front group, and highly-differentiated tissue was more common in non-front group(χ²=6.88, P=0.03). There were significant differences in the expression levels of β-tubulin Ⅲ between cancer tissues with different differentiation degrees (χ²=5.74, P=0.04). In non-front group, lymph node metastasis was closely correlated with the expression of β-tubulin Ⅲ (χ²=6.02,P=0.05). The resultsof immunohistochemical staining showed that the β-tubulin Ⅲ positive-expressing cells were colored brown-yellow. The number of cells with positive β-tubulin Ⅲ expression was significantly increased in highly differentiated tissue compared with low-differentiated tissue.Conclusion: The expression of β-tubulin Ⅲ is closely related to tumor differentiation in colon adenocarcinoma tissue. The highly differentiated colon adenocarcinoma tissue is more common in non-front group in which the expression of β-tubulin Ⅲ is related to lymph node metastasis.

Objective: To observe the influence of the CD96 and CD71 expressions in the surface of leukemia stem cells (LSC) in the therapeutic effects and prognosis of the children with acute leukemia,and to clarify the relationships between the molecular biological characteristics of LSC in the children with leukemia and their therapeutic effects and prognosis.Methods: Eighty children with acute leukemia were selected as the subjects. Among them, 39 cases were acute lymphoblastic leukemia (ALL) and 41 cases were acute myeloid leukemia (AML). The CD96 and CD71 expressions on the surface of LSC were detected with flow cytometry; the therapeutic effects of the first cycle chemotherapy, the survival rate of 5-year, the incidence of infection after chemotherapy, the recurrence rate after chemotherapy, and the incidence of extramedullary infiltration of the children were observed and compared.Results: The CD96 expression on the surface of LSC was positive in 38 cases (47.5%) and the CD71 expression on the surface of LSC was positive in 45 cases (56.3%);the difference of positive expression rates of CD96 and CD71 was not significant (χ²=1.227, P=0.268). The positive expression rates of CD96 and CD71 on the surface of LSC of the children with AML were significantly higher than those in the children with ALL, and the differences were statistically significant (χ²=22.225, χ²=34.028, P<0.01). The distribution of therapeutic effects and the clinical efficiency of the first cycle chemotherap, in the children with negative CD96 expression on the surface of LSC were superior to those with positive CD96 expression on the surface of LSC; the distribution of therapeutic effects and the clinical efficiency of the first cycle chemotherapy in the children with negative CD71 expression on the surface of LSC were superior than those with positive CD71 expression on the surface of LSC; the differences between two groups were statistically significant (χ²=11.323, χ²=16.589, P<0.05; U=2.939, U=2.291, P<0.05). The survival rate of 5-year in the children with negative CD96 expression on the surface of LSC was higher than those with positive CD96 expression; the incidence of infection after chemotherapy,the recurrence rate after chemotherapy and the incidence of extramedullary infiltration were lower than those with positive CD96 expression. The incidence of infection after chemotherapy and the recurrence rate after chemotherapy in the children with negative CD71 expression on the surface of LSC were lower than those with positive CD71 expression,and the differences between two groups were statistically significant (χ²=5.051,χ²=13.288, P<0.05).Conclusion: The expressions of CD96 and CD71 on the surface of LSC in the children with acute leukemia has relationship with the subtypes of the disease and the therapeutic effects of the first cycle chemotherapy, which can be used as markers for the diagnosis and evaluation of therapeutic effects. The expression level of CD96 is related to the prognosis of the patients, which can be used as an indicator for predicting the prognosis of patients.

Objective: To investigate the role of interleukin-10 (IL-10) and IL-12 in the differential diagnosis of tuberculous and malignant pleural effusion through measuring the serum and pleural effusion IL-10 and IL-12 levels of tuberculous and malignant pleural effusion patients.Methods: Forty-eight inpatients with exudative pleural effusion who didn't receive any treatment before were selected. According to the pathogeny, the patients were divided into tuberculous pleural effusion group (n=25) and malignant pleural effusion group (n=23). The levels of IL-10 and IL-12 in peripheral blood and pleural effusion of the patients in two groups were detected by Cytometric Bead Array (CBA). The differences in IL-10, IL-12 levels and IL-12/IL-10 ratio were compared between two groups. The ROC curves of the above indexes, cast-off and adenosine deaminase (ADA) were used to compare the effects in differential diagnosis of tuberculous and malignant pleural effusion.Results: There were no statistical differences in the levels of serum IL-10, IL-12 and IL-12/IL-10 ratios of the patients between tuberculous and malignant pleural effusion groups (P>0.05),and the IL-12 level in pleural effusion and the IL-12/IL-10 ratio of the patients in malignant pleural effusion group were lower than those in tuberculous pleural effusion group (P<0.005).The area under the ROC curve for the levels of IL-12 in pleural effusion (0.984) was higher than those for the ratio of IL-12/IL-10, cast-off and ADA (0.744, 0.804 and 0.911, respectively) in the differential diagnosis of tuberculous and malignant pleural effusion.Conclusion: The detection of IL-12 levels in pleural effusion is helpful for the differential diagnosis of tuberculous and malignant pleural effusion, and the pleural effusion is easy to obtain, therefore the detection of IL-12 in pleural effusion is worth to be widely applied in clinic.

Objective: To investigate the efficacy and the clinical value of magnetic resonance elastography(MRE) in diagnosis of hepatic fibrosiswith Meta-analysis, and to provide basis for clinical treatment of hepatic fibrosis.Methods: The studies published before February 2, 2017 about MRE and staging of hepatic fibrosis in Chinese or English were retrived in the databases including PubMed, EMBase, Web of Science, Cochrane Library,CNKI, CBMDisc,VIP, Wanfang data, and supplemented by manual retrieval for relevant literatures. The inclusion and exclusion criterions were used to select and extract the literatures.The literatures qualitie were valuated based on QUADAS-2 tool.The sensitivity(SEN), specificity (SPE), diagnostic odds ratio (DOR), positive likelihood ratio (+LR), negative likelihood ratio (-LR) on the groups of F0 vs F1-F4,F0-F1 vs F2-F4,F0-F2 vs F3-F4, F0-F3 vs F4 and heterogeneity were combined and tested with Stata software respectively.HSROC and AUROC were also implemented.Results: A total of 1 332 studies were searched, and 22 were included. 21 of them were in English and 1 in Chinese. The results of Meta analysis showed that the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0 vs F1-F4 group were 88.8%(85.0-91.7),95.9%(91.5-98.0),21.435(10.215-44.979),0.117(0.086-0.159),183.187(72.533-462.650) and 0.96(0.94-0.98),respectively;the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0-F1 vs F2-F4 group were 93.3%(89.2%-35.9%), 94.1%(90.2%-96.5%),15.839(9.344-26.848),0.072(0.044-0.117),221.224(100.980-484.648) and 0.98(0.96-0.99),respectively; the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0-F2 vs F3-F4 group were 92.9%(88.9%-95.5%),94.6%(91.2%-96.8%),17.348(10.496-28.671),0.075(0.048-0.119),230.434(111.482-476.317)0.98(0.96-0.99), respectively;the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0-F3 vs F4 group were 97.7%(93.0%-99.3%),93.2%(90.3%-95.2%),14.337(9.910-20.742),0.025(0.008-0.075),580.405(144.871-2325.307) and 0.98(0.96-0.99),respectively.Conclusion: MRE,as a new and noninvasive imaging method, has high diagnostic value in all stages of hepatic fibrosis, which can provide a reliable reference for clinical precise treatment of hepatic fibrosis.

Objective: To establish the quantitative relationshipequationof the crantiofacial vertical points in the skeletal classⅡ malocclusion patients with various vertical types by using genetic algorithms method,and to express the measured values in the patients with different gender with the same formula.Methods: A total of 155 skeletal class Ⅱ malocclusion patients without treatment,aged from 10 to 18 years old,were selected and divided into high-angle group(n=50),average-angle group(n=58),low-angle group(n=47);5 samples were randomly selected in each group as the test samples,the rest as the experimental sample.The cephalometic radiographs were performed and measured.The relevant influencing factors of craniofacial structure were ensured.The genetic algorithm was used to optimize the equation parameters to obtain the correlation equation.The error between the predicted value and the measured value was compared.Results: The various parameters had no significant differences between different gender in high-angle,average-angle and low-angle groups(P>0.05);then the men and the women with same type were combined,most of the indicators had statistically significant differences between three groups (P<0.05).The correlation analysis results showed that there was a positive correlation between age and Ans-U1(r=0.470),there was a positive correlation between N-Me and Ans-Me(r=0.964);for Ans-U1,there was a positive correlation with Ans-Me(r=0.805)and negative correlation with facial angle;there was a positive correlation between N-Go and N-Me (r=0.926);for L1-Me,there was a positive correlation with Ans-Me(r=0.898)and negative correlation with the angle of Go(r=-0.468)in high-angle group. In average-angle group,there was a positive correlation between age and N-Me (r=0.531);for Ans-U1,there was a positive correlation with Ans-Me(r=0.878)and negative correlation with the facial angle(r=-0.262);for Ans-Me,there was a positive correlation with N-Me(r=0.920), negative correlation with N-Ans(r=-0.560)and negative correlation with Ar-Go(r=-0.652);for N-Go,there was a positive correlation with S-Go(r=0.867), positive correlation with N-Ans(r=0.252)and positive correlation with L1-Me(r=0.754).For S-Ar,there was a positive correlation with S-Go(r=0.671), negative correlation with Ar-Go(r=-0.250),and positive correlation with L1-Me(r=0.552).In low-angle group,for age,there was a positive correlation with S-Go(r=0.602), negative correlation with the angle of Go(r=-0.346),and positive correlation with L1-Me(r=0.576);for N-Me,there was a positive correlation with Ans-Me(r=0.869),and positive correlation with N-Go(r=0.859),and negative correlation with the facial angle(r=-0.177);for N-Ans,there was a positive correlation with N-Me(r=0.605) and negative correlation with Ans-U1(r=-0.113);for Ans-Me,there was a positive correlation with N-Me(r=0.869),positive correlation with the facial angle(r=0.070),and positive correlation with Ans-U1(r=0.785);for N-Go,there was a positive correlation with N-Me(r=0.859)and positive correlation with S-Go(r=0.829).The quantitative relationship equations of the crantiofacial vertical points in skeletal class Ⅱmalocclusion patients with various vertical types in each group were established by using genetic algorithms.In high-angle group:Age=5.883 6+0.269×Ans-U1,N-M=22.026 6+1.494 5×Ans-Me,Ans-U1=34.959 4+0.454 5×Ans-Me-0.409 7×Facial angle,N-Go=-4.588 2+0.472 4×N-Me,L1-Me=-12.590 5+0.5322×Ans-Me+0.124 3×∠Go.In average-angle group:Age=-2.944 1+0.146 8×N-Me,Ans-U1=18.917+0.476 4×Ans-Me-0.230 2×Facial angle,Ans-Me=-0.620 5+1.014 5×N-Me-0.974 1×N-Ans-0.057 6×Ar-Go,N-Go=1.631 1+0.897 8×S-Go+ 0.919 7×N-S+0.168 8×L1-Me,S-Ar=-1.823 1+ 0.845 3×S-Go-0.867 0×Ar-Go+0.202 4×L1-Me.In low-angle group:Age=11.740 6+0.152 7×S-Go-0.169 9×∠Go+0.252 5×L1-Me,N-Me=61.153 0+0.964 3×Ans-Me+0.628 6×N-Go-0.689 2×Facialangle,N-Ans=-4.949 2+1.065 8×N-Me-2.316 5×Ans-U1,Ans-Me=-25.180 0+0.418 4×N-Me+0.280 3×Facial angle+0.477 6×Ans-U1,N-Go=8.684 2+0.409 9×N-Me+0.403 3×S-Go.There was no significant difference between the predicted values of equation established with genetic algorithms and the measured data (P>0.05).Conclusion: The quantitative relationship equation of the crantiofacial vertical points in the skeletal class Ⅱmalocclusion patients with various vertical types established with genetic algorithms may show the vertical quantitative relationship and predict the growth to a certain degree.

Objective: To optimize the parameters of the equation of sagittal craniofacial structures with different classes of malocclusion using genetic algorithms(GAS), and to explore the rules.Methods: Atotal of 240 patients with average angle malocclusion aged 8-18 years old were divided into three groups:Angle Class Ⅰ(n=79), Angle Class Ⅱ(n=76)and Angle Class Ⅲ(n=85) groups.In each group 10 cases were randomly selected as the test samples, the rest as the experimental samples.The cephalometric analysis was performed on all the patients' cephalograms, and the results of Ba-N,Ba-A,Ba-S,S-Ptm,Ptm-A,Ba-Ar,Ar-Go,Go-PoG,Ba-PoG and N-S-Ar were analyzed by two independent samples t-test and One-Way ANOVA. The relevant influencing factors of craniofacial structures were found.The parameters of the equation was optimized to obtain the relevant equations using GAS.The predicted values of the optimized equation were compared with the measured values.Results: There were no significant differences in sex between Angle Class Ⅰ, Class Ⅱ and Class Ⅲ groups(P> 0.05);when the men and women with the same type were combined,the Ba-A,Ptm-A,Ar-Go,and Ba-PoG had statistically significant differences between Angle Class Ⅰ, Class Ⅱ, and Class Ⅲ groups (P<0.05).The correlation analysis results showed that in Angle Class Ⅰgroup:Ba-A was positively correlated with Ba-N (r=0.683),Ptm-A was positively correlated with Go-PoG (r=0.738), Ar-Go was positively correlated with Ba-PoG (r=0.833), and negatively correlated with Go-PoG (r=-0.560) and Ba-PoG was positively correlated with Go-PoG (r=0.669); in Angle class Ⅱ group,Ba-A was positively correlated with Ba-PoG and Ba-N(r=0.884,r=0.883), Ptm-A was positively correlated with Ba-A (r=0.742),Ar-Go was positively correlated with Ba-PoG (r=0.401)and negatively correlated with Go-PoG (r=-0.317) and Ba-PoG was positively correlated with Ba-A and Go-PoG(r=0.883,r=0.488);in Angle Class Ⅲ group,Ba-A was positively correlated with Ba-N and Ba-PoG(r=0.891,r=0.829),Ptm-A was positively correlated with Ba-A (r=0.807)and negatively correlated with Ba-S (r=-0.404),Ar-Go was positively correlated with S-Ptm (r=0.548) and Ba-PoG was positively correlated with Ba-A (r=0.829).The equation of sagittal craniofacial structure with different occlusal classes was established by GAS.In Angle Class Ⅰgroup:Ba-A(mm)=10.963 9+0.859 8×Ba-N,Ptm-A(mm)=6.897 6+0.557 0×Go-PoG,Ar-Go(mm)=-2.548 2+ 0.511 8×Ba-PoG-0.5272×Go-PoG,Ba-PoG(mm)=17.515 6+1.021 3×GO-POG;in Angle Class Ⅱ group:Ba-A(mm)=-2.121 3+0.567 6×Ba-PoG+0.513 2×Ba-N,Ptm-A(mm)=13.788 7+0.349 4×Ba-A,Ar-Go(mm)=2.447 7+0.368 8×Ba-PoG-0.427 9×Go-PoG,Ba-PoG(mm)=-7.140 2+0.751 3×Ba-A+0.295 4×Go-PoG;in Angle Class Ⅲgroup:Ba-A(mm)=3.281 0+0.545 3×Ba-N+0.394 4×Ba-PoG,Ptm-A(mm)=3.535 8+0.63 1×Ba-A-0.614 2×Ba-S,Ar-Go(mm)=-9.002 1+1.004 3×S-Ptm,Ba-PoG(mm)=-2.091 2+1.057 5×Ba-A.There were no significant differences between the predicted values of GAS and the measured data (P> 0.05), and the error was small.Conclusion: The optimal relation equation of craniofacial structure of sagittal malocclusion is established by GAS with the quantitative regularity.

Objective: To investigate the relationship between primary resection and KRAS gene mutation in the mild symptomatic patients with stage Ⅳ colorectal cancer,and to clarify its significance of prognosis.Methods: The clinical data of 46 mild symptomatic patients with stage Ⅳ colorectal cancer 2010 to December 2010 were collected. All the patients received primary resection.The KRAS gene mutation in the patients was detected by direct sequencing and the patients were followed up for 5 years.The influence of primary resection and KRAS gene mutation in prognosis of the patients with stage Ⅳ colorectal cancer was analyzed, and the clinical pathological features which might influence the prognosis were analyzed by survival analysis.Results: In 46 patients with colorectal cancer, KRAS gene mutation was found in 20 cases, the mutation rate was 43.4%, and most mutation was found at Codon 12. The KRAS mutation had relationship with the tumor site and multiple metastasis (P<0.05).The Kaplan-Meier survival curve and univariate analysis results showed that the median survival time of the patients with wild type of KRAS gene was 58.4 months, the median survival time of the patients with mutant type of KRAS gene was 42.2 months, and there was no significant difference (P>0.05).The median survival time of right colon cancer patients was 34.2 months, the median survival time of left colon cancer patients was 58.3 months, and there was sigificant difference (P<0.05).The cancer metastases including liver, lung and multiple metastasis were closely related to the poor prognosis of the colorectal cancer patients(P<0.05).The median survival time of the patients with stage Ⅳ colorectal cancer was 39.6 months after operation.Conclusion: After primary resection of the mild symptomatic patients with stage Ⅳ colorectal cancer,the median survival time of the patients with colorectal cancer in left colon site and right colon site were prolonged. Right colon cancer has more poorer prognosis than left colon cancer. KRAS gene mutation is associated with the tumor site and the multiple metastasis. The location of metastasis affect the prognosis.

Objective: To explore the curative effect and toxic and side effects of Xihuang Capsule in the patients with advanced esophageal cancer treated with chemoradiotherapy, and to clarify its mechanism.Methods: The patients with advanced esophagealcancer without treatment were selected. The patients treated with Xihuang Capsule combined with concurrent chemoradiotherapy were defined as combination treatment group(n=27), and then other 27 patients were randomly selected from 87 patients treated with concurrent chemoradiotherapy alone as chemoradiotherapy group. The observation time was 90 d from the begining of chemotherapy. The differences in KPS, degree of dysphagia, incidence, occurrence time and degrees of acute radiation esophagitis of the patients in two groups were compared, and local control of tumor was analyzed.Results: After treatment, the KPS scores of the pateints in two groups were decreased, and the KPS score in combined treatment group was significantly higher than that in chemoradiotherapy group (P<0.05). The grade of dysphagia of the patients in combined treatment group was lower than that in chemoradiotherapy group during the late period of treatment(P<0.05). The incidence of acute radiation esophagitis, the occurence time of acute radiation esophagitis of the patients in combination group was lower than that in chemoradiotherapy group (P<0.05);the occurrence time of acute radiation esophagitis was later than that in chemoradiotherapy group (P<0.05); but there were no significant differences in the grades of acute radiation esophagitis and tumor local control between two groups(P>0.05).Conclusion: Xihuang Capsule can improve the quality of life, reduce dysphagia degree and the incidence of acute radiation esophagitis,and delay the occurrence time;it can be used as an effective auxiliary treatment of advanced esophageal cancer.

Objective: To observe the clinical application of flexible endoscope assisted by general versus pillow-under-shoulder supine position in nasotracheal intubation of the patients with difficult airway, and to explore the influence of intubation position in the intubation effect.Methods: A total of 168 patients with difficult airway who underwent nasotracheal intubation and oromaxillofacial surgery under general anesthesia were randomly divided into general supine position (control group) and pillow-under-shoulder supine position (experimental group) with 84 cases in each group. The first-time and the total success rate of intubation, the intubation time, and the rate of direct glottis exposure of the patients in two groups were recorded. The mean arterial pressure(MAP), heart rate (HR), and complications of intubation of the patients in two groups before induction, before tracheal intubation, during intubation, 1 and 5 min after intubation, were also recorded.Results: The first-time success rate of intubation in experimental group (94.0%, 79/84) was significantly higher than that in control group (71.4%, 60/84) (P<0.01); the total success rate of intubation (98.8%, 83/84) had no difference compared with control group (97.6%, 82/84) (P>0.05); the intubation time (57 s±12 s) was significantly shorter than that in control group (146 s±29 s) (P<0.01); the rate of direct glottis exposure (47.6%, 40/84) when the flexible endoscope passed through the posterior nasal apertures was obviously higher than that in control group (15.5%, 13/84) (P<0.01).The values of MAP and HR of the patients in two groups before and during intubation were decreased significantly compared with the baselines (P<0.01), and came back to the baselines 1 and 5 min after intubation (P>0.05). There were no significant differences in the MAP and HR between different time points (P>0.05). The incidence rates of complications including pharyngalgia, hoarseness and epistaxis had no differences between two groups (P>0.05).Conclusion: Flexible endoscope assisted by pillow-under-shoulder in nasotracheal intubation has a higher intubation success rate, shorter intubation time and it is a superior procedure for the patients with difficult airway.

Objective: To treat the mixed and deeper infantile hemangiomawith "ladder dosage"propranolol, and to explore its efficacy and safety.Methods: A total of 98 infants with hemangioma were treated by ladder treatment of propranolol. Before treatment,comprehensive assessments of electrocardiogram(ECG),heart color ultrasound, blood glucose,liver function,kidney function,myocardial enzymes and blood routine were conducted. After excluding contraindications,the dose of propranolol incrementally doubled from 0.5 mg·kg^-1·d^-1 to 4.0 mg·kg^-1·d^-1. Propranolol was taken 3 times a day. Before and after medication for 1 and 2 h,ECG was monitored. The changes of tumor size,texture,color and other changes or an onset of adverse reactions were dynamicly observed.The infants were visited every month.The efficacy was evaluated using Achauer system.Results: After medication,98 cases had different degrees of color changes or tumor consistency softening. After the dosage of propranolol was increased to 4.0 mg·kg^-1·d^-1,the change of tumor was the fastest. According to the 4-grade method, there were 84 cases(85.71%) as gradeⅣ (excellent),2 cases (2.04%) as grade Ⅲ (good),4 cases (4.08%) as gradeⅡ (medium)and 8 cases (8.16%) as gradeⅠ (poor).The curative effect of mixed hemangioma was better than that of deeper hemangioma(P<0.05). The recovery time of 74 cases of hemangiomas was 6 months. The major adverse reactions were heart rate decline(5/98,5.10%),drowsiness(3/98,3.06%),diarrhea(7/98,7.14%),loss of appetite (1/98,1.02%), and convulsions (2/98,2.04%).After treatment,all adverse reactions disappeared. Two months after drug withdrawal there were 4 cases of recurrence,and they were continously treated with propranolol.Conclusion: The efficacy of oral "ladder dosage" propranolol in treatment of mixed and deeper infantile hemangioma is increased significantly and there are no significant adverse reactions.

Objective: To summarize the therapeutic effect of free deep inferior epigastric perforator (DIEP) flap and anterolateral thigh perforator (ALTP) flap in repairing skin and soft tissue defects in chest wall tumor resection.Methods: One old male patient with malignant fibrous histiocytoma of chest wall in China-Japan Union Hospital received operation,and the tumor was extendedly resected.The remaining chest wall defects were 10 cm×10 cm and 18 cm×14 cm,respectively.Free DIEP and ALTP flaps were used to repair the defects,and vascular pedicle was anastomosed with internal mammary artery and thoracic lateral artery,respectively.Results: After operation,the donor area was sutured directly,and two flaps survived with good appearance. The skin of ALTP flap was flatter than that of DIEP flap,and the texture and color of skin were close to those around chest wall.Conclusion: Free DIEP and ALTP flaps are ideal choice for repairing skin and soft tissue defects after resection in the male patient with chest wall tumor.

Objective: To report one case of renal clear cell carcinoma transferred to the nasal cavity and sinuses treated by low temperature coblation assisted nasal malignant tumor resection under nasal endoscopye, and to review the associated literatures.Methods: Under nasal endoscope, the tumor was removed along the lateral wall of the nasal cavity with bipolar electrocautery and coblation. The vaseline gauze was used to stop bleeding, and the tumor was completely removed.Results: The intraoperative frozen section showed a capillary hemangioma. The patient was diagnosed as metastatic renal cell carcinoma to the nasal cavity and sinuses by the results of pathology and immunohistochemical staining. The postoperative recovery was uneventful and the nasal packing was removed 3 d after operation.Conclusion: Renal clear cell carcinoma transferred to the nasal cavity and sinuses is rare; under nasal endoscope, surgical resection is a good procedure to control metastasis.

Objective: To investigate the clinical manifestations and distribution characteristics of dry eye patients with ocular discomfort symptoms in Jilin Province, and to analyze the relationship between the risk factors associated with dry eye and its severity.Methods: The secondary or tertiary hospitals in Jilin Province were randomly selected and used as screening bases from July 2014 to August 2015. 1 173 people initiative to the hospital for eye examinations after publicity were selected. Questionnaire was used to collect the subjective symptoms of dry eye. The breakup time(BUT) of tear film, corneal fluorescein staining, meibomian gland and tear secretion were examined and the detection rate and risk factors of dry eye of the dry eye patients with different clinical characteristics were analyzed.Results: A total of 1 122 people was actually surveyed, 896 individuals were diagnosed as dry eye, and the prevalence rate was 79.8%. The prevalence rate of the females was higher than of the males(χ²=4.070,P<0.05).The prevalence of dry eye between different ages was statistically significant(χ²=61.547,P<0.05).The multivariate Logistic regression analysis results showed that the age ≥ 40 years (40-49years,OR=6.313,95% CI:3.498-11.393; 50-59 years,OR=6.919,95% CI:3.876-12.351; 60-69 years,OR=5.175,95% CI:2.650-10.104; over 70 years,OR=9.508,95% CI:3.608-25.061), moderate grade of meibomian gland dysfunction(MGD) (OR=2.123,95% CI:1.186-3.803), and the patients with rheumatoid arthritis (OR=2.186,95% CI:1.098-4.353) and eye surgery (OR=3.692,95% CI:1.204-11.323) were the risk factors for dry eye. While the occupation of farmer was a protective factor for dry eye (OR=0.351, 95% CI:0.135-0.917).Conclusion: Age, occupation, MGD grade, rheumatoid arthritis and eye surgery history affect the occurrence of dry eye to a certain extent. So enough attention and appropriate health guidance should be given to reduce the incidence of dry eye.

Objective: To prepare the multilayer alginate chitosan microspheres loading vascular endothelial growth factor (VEGF) and vancomycin (VAN), and to study in vitro release characteristics.Methods: The microspheres were prepared by emulsion cross-linking and self-assembly techniques. The effects of sodium alginate concentration, calcium chloride concentration, oil/water ratio and span80 concentration on the entrapment efficiency(EE) and drug loading(DL) of VEGFand VAN were investigated by orthogonal experimental design to optimize the preparation process. The surface morphology and particle size of microspheres were observed by scanning electron microscope (SEM). Self-assembly was detected by Fourier transform infrared spectroscope (FTIR). The EE, DL and in vitro release of VEGF and VAN were detected by ELISA double antibody sandwich method and ultraviolet spectrophotometry,and the cumulative release curve was drawn.Results: The prepared microspheres were yellowish brown powder. The SEM results showed that the microspheres were spherical, the surface was smoothy, and the dispersity was better. The average particle size was about 50 μm. Sodium alginate concentration of 1.0g·mL^-1, CaCl₂ concentration of 8g·mL^-1, oil to water ratio of 3:1, and span80 concentration of 2% were the best formula. The EE of VEGF and VAN were 49.63% and 16.67%, respectively. In vitro, the cumulative release last 16.5 d and 12.5 d respectively and the amount reached up to 95%.Conclusion: The multilayer alginate chitosan microspheres loading VEGF and VAN present several advantages, such as smaller particle size, higher EE and better controlled release.