Objective: To investigate the influence of arsenic troxide(As₂O₃) and γ-secretase inhibitor MW167 used alone or in combination in the drug resistance of HepG/ADM cells,and to clarify its mechanisms. Methods: The HepG₂/ADM cells were treated with different concentrations of As₂O₃(0.25,0.50,1.00,2.00,4.00,8.00 mg·L^-1) and MW167(10,20,40 μmol·L^-1) for 24,48 and 72 h,and MTT method was used to detect the optical density (A) values,then the inhibitory rates of proliferation were calculated and the drug concentration and treatment time were confirmed.The HepG₂/ADM cells were divided into blank group,As₂O₃ group, MW167 group and combination group; the non-cytotoxic doses of As₂O₃ and MW167 according to the results of MTT were chosen and the HepG₂/ADM cells were intervened for 48 h; the apoptotic rates were detected by FCM; RT-PCR and Western blotting methods were employed to detect the mRNA and protein expression levels of Notch-1,Hes-1,MDR-1 (P-gp),Bcl-2 and Bax in the cells in various groups. Results: At the same concentration,the inhibitory rates of proliferation of HepG₂/ADM cells were enhanced with the prolongation of the treatment time of As₂O₃ and MW167(P<0.05 or P<0.01); at the same time point,the inhibitory rates of proliferation of As₂O₃ and MW167 in the HepG₂/ADM cells were increased with the increasing of drug concentration (P<0.05); the non-toxic doses of As₂O₃ and MW167 were 0.25 mg·L^-1 and 10 μmol·L^-1 and the intervention time was 48 h.After treatment for 48 h,compared with blank group,the apoptotic rates in various intervention groups were increased (P<0.05),especially in combination group (P<0.01); the expression levels of Notch-1,Hes-1,MDR-1 (P-gp),Bcl-2 mRNA and protein in various intervention groups were lower than those in blank group (P<0.05),especially in combination group (P<0.01);compared with blank group,the expression levels of Bax mRNA and protein in various intervention groups were increased (P<0.05),especially in combination group (P<0.01). Conclusion: As₂O₃ can reverse the drug resistance of HepG₂/ADM cells,its mechanism may be related to inhibition of cell proliferation,promotion of apoptosis,down-regulation of the expression levels of Notch-1,Hes-1,MDR (P-gp),Bcl-2 and up-regulation of the expression levels of Bax mRNA and protein.

Objective: To explore the effects of chronic restraint stress (CRS) on the abilities of spatial learning and memory and the levels of excitatory amino acids in the hippocampal dentate gyrus (DG) in the old rats,and to investigate the neurochemical mechanism of CRS in affecting the spatial learning and memory abilities. Methods: Sixteen male SD rats (18 months old) were randomly divided into control group (n=8) and CRS group (n=8),and the rats in CRS group received CRS 2 h every day for 30 d.And then the spatial learning and memory abilities of rats were measured by Morris water maze(MWM) test,and the extracellular levels of excitatory amino acids including asparate (Asp) and glutamate (Glu) in the DG were simultaneously determined by in vivo microdialysis and HPLC.The levels of corticosterone (CORT) and epinephrine (EPI) in serum of the rats were examined by ELISA assay. Results: In CRS group,the escape latencies on the 2nd-4th days were significantly increased and the percentage of time spent in target quadrant on the 5th day was markedly decreased in MWM test compared with control group (P<0.05).Compared with before training,the extracelluar level of Asp in the DG in control group was significantly increased on the 2nd day in MWM test;compared with control group,the extracelluar level of Asp in the DG in CRS group was significantly decreased on the 3rd day in MWM test(P<0.05).Compared with before training,the Glu levels in the DG in MWM test in both control and CRS groups were markedly increased (P<0.05),but there was no significant difference between two groups (P>0.05).Compared with control group,the levels of CORT and EPI in the serum of the rats in CRS group were significantly increased (P<0.05). Conclusion: CRS impairs the spatial learning and memory abilities in the old rats,which may be related to the decrease of Asp level in the hippicampal DG of the rats.

Objective: To explore the inhibitory effect of Shiquandabu decoction on the spontaneous len tumor of the SV40 T antigen transgenic (TG) mice,and to clarify its molecular mechanism. Methods: The SV40 T antigen TG mice were randomly divided into control group (n=39) and drug treatment group (n=25).The mice in control group were fed normally,while the mice in drug treatment group were fed with Shiquandabu decoction at the 3rd week after birth,the survival time of mice was recorded.Three mice in control group and drug treatment group were randomly chosen to collect the blood from the tail vein and the amino acid levels were measured respectively 8 and 15 weeks after Shiquandabu decoction administration.Then the mice were sacrificed and the liver tissue was collected.Gene chip hybridization was used to detect the differences in the expressions of ribosomal function related genes in liver tissue of the mice in two groups and the related signal pathway was explored. Results: The survival analysis demonstrated that the survival rate of TG mice in drug treatment was higher than that in control group (P<0.05).Compared with control group,the serum levels of alanine,valine,leucine,isoleucine,threonine,methionine,proline,tyrosine,lysine,sarcosine,citrulline,ornithine and hydroxylysine of the mice in drug treatment group 8 weeks after administration of Shiquandabu decoction were increased (P<0.05); and the serum levels of cystathionine,taurine,methylhistidine,anserine and ethanolamine were decreased (P<0.05).Fifteen weeks after administration,compare with control group,the serum levels of threonine and citrulline of the mice in drug teeatment group were increased (P<0.05),but the serum levels of other amino acids had no significant difference (P>0.05).The canonical analysis showed that thirteen genes involved in ribosomal function from 9 083 genes in liver tissue in drug treatment group had the changes compared with control group(P<0.05). Conclusion: Shiquandabu decoction can effectively prolong the lifetime of the TG mice by improving the levels of serum amino acids and promoting the liver ribosomal protein synthesis.

Objective: To study the inhibitory effects of andrographolide(Andro) on the proliferation,migration and clone formation ability of renal cell carcinoma(RCC) cells and the induction on the apoptosis,and to clarify their related mechanisms. Methods: The RCC cells were treated with different concentrations (5,10,20,40 and 80 μmol·L^-1) of Andro as experimental groups,and 0 μmol·L^-1 Andro group was used as blank control group,MTS assay was used to detect the proliferation rates of RCC cells in various groups.The RCC cells were treated with different concentrations (0.50,1.25 and 2.50 μmol·L^-1) of Andro as experimental groups,and 0 μmol·L^-1 Andro group was used as blank control group.Clonogenic assays was used to detect the colony formation ability of RCC cells in various groups. The RCC cells were treated with different concentrations (10,20 and 40 μmol·L^-1) of Andro as experimental groups,and 0 μmol·L^-1 Andro group was used as blank control group.Wound healing assay was used to detect the migration ability of RCC cells in various groups.Flow cytometry was used to detect the apoptotic rates of RCC cells in various groups.Western blotting was performed to detect the expression levels of apoptosis related proteins in RCC cells in various groups. Results: Compared with blank control group,the proliferation rates of RCC cells in 10,20,40 and 80 μmol·L^-1 Andro groups were markedly decreased(P<0.05 or P<0.01).Compared with blank control group,the colony formation rates of RCC cells in 0.50 and 1.25 μmol·L^-1 Andro groups were markedly decreased(P<0.05 or P<0.01).Compared with blank control group,the scratch healing rates of RCC cells in 10,20 and 40 μmol·L^-1 Andro groups were markedly decreased (P<0.01),and the apoptotic rates of RCC cells in 20 and 40 μmol·L^-1 Andro groups were markedly increased (P<0.01).Compared with blank control group,the expression level of γ-H2AX protein in 40 μmol·L^-1 Andro group was markedly increased(P<0.01),the expression level of Caspase-8 protein was decreased (P<0.05), and the expression level of cleaved Caspase-8 protein was markedly increased(P<0.01). Conclusion: Andro can effectively suppress the proliferation,migration and colony formation ability of RCC cells and induce the apoptosis of RCC cells.The mechanism of apoptosis might be related to inducing the DNA damage and the apoptotic pathways induced by JNK/H2AX and Caspase-8.

Objective: To study the effect of isoflurane anesthesia on the hippocampal neuroapoptosis in the zinc-deficient APP/PS1 transgenic mice,and to clarify its related mechanisms. Methods: The eight-month or nine-month old APP/PS1 transgenic mice were randomly assigned to four groups (n=24):zinc adequate(ZA) group,the mice were given a standard diet for 2 months; zinc adequate+ isoflurane(ZA+Iso) group,the mice were given a standard diet for 2 months then exposed to 1.4% isoflurane for 2 h; zinc-deficient(ZD) group,the mice were given zinc deficient diet for 1 month; zinc deficient with isoflurane(ZD+Iso)group,the mice were given zinc deficient diet for 1 month and then exposed to 1.4% isoflurane for 2 h. The hippocampus tissue of the mice were obtained 24 h after anesthesia. The immunofluorescence double staining was performed to measure the apoptotic rates of hippocampal neurons. The Western blotting method was used to detect the expression levels of Aβ and Cleaved caspase-3 and the ratio of Bax/Bcl-2. Results: Compared with ZA group,the apoptotic rate of neurons,the ratio of Bax/Bcl-2 and the expression level of Cleaved caspase-3 of the mice in ZA+Iso group were increased 6 h after isoflurane exposure(P<0.05 or P<0.01);but no differences were found in the apoptotic rate of neurons and the expression levels of total Aβ,Aβ40 and Aβ42 24 h after isoflurane exposure(P>0.05).Compared with ZA group,the apoptotic rates of neurons,the ratios of Bax/Bcl-2,the expression levels of Aβ42 and Cleaved caspase-3 of the mice in ZD group and ZD+Iso group were increased 6 and 24 h after isoflurane exposure(P<0.05 or P<0.01).Compared with ZD group,the apoptotic rate of neurons,the expression levels of Aβ42,Bax/Bcl-2 and Cleaved caspase-3 and the ratio of Bax/Bcl-2 of the mice in ZD+Iso group were elevated significantly 6 and 24 h after isoflurane exposure(P<0.05 or P<0.01). Conclusion: 1.4% isoflurane exposure for 2 h can induce the transient elevation of hippocampal neuroapoptosis in the ten-month old APP/PS1 transgenic mice.1.4% isoflurane exposure for 2 h can significantly aggravate the hippocampal neuroapoptosis in the zinc-deficient APP/PS1 transgenic mice,it is probably associated with the hippocampal Aβ aggregation,activation of Bax and Caspase-3 and inhibition of Bcl-2.

Objective: To explore the effects of the activating circulation to remove blood stasis through observingthe influence of Inonotus hispidus on hemorheology in the rat models with blood stasis due to cold syndrome.. Methods: The rats were randomly divided into normal control group, blood stasis model group, positive control group, low and high doses ofInonotus hispidus(S1-S6) sample groups; there were 15 groups,and 12 rats were in each group. Except for normal control group, the rats in other groups were injected adrenaline hydrochloride and immersing in ice water to build the blood stasis due to cold syndrome rat models.The indexes of hemorheoloy of the rats were detected,and the HPLC spectrum-effect relationship was analyzed. Results: The yellow fruiting bodies(S3-S6) of Inonotus hispidus which were parasitic on the different trees significantly improved the hemorheological indexes in the rat models.Compared with model group, the blood viscosity and whole blood viscosity of the rats in high dose of yellow fruiting body of Inonotus hispidus group were decreased(P<0.05).The black fruiting body of Inonotus hispidus showed the similar effect to the yellow fruiting body, but the hemorheological indexes of the rats in black fruiting body of Inonotus hispidus group had no significant changes compared with model group(P>0.05). The results of the spectrum-effect showed that the corresponding components of 11 chromatographic peaks were closely correlated with the changes of plasma viscosity, and the correlation coefficient > 0.8.Through identification,four compounds of them were named phellibaumin A,phelligridin C(cis),phelligridin C(trans),phelligridin D and 3'4'-dihydroxy-5-[(11-hydroxyphenyl)-6,7-vinyl]-3,5-dioxa-fluoren-5-one. Conclusion: The yellow fruiting bodies of Inonotus hispidus which are parasitic on the different trees can decrease the blood viscosity and whole blood viscosity and have the effects of activating the circulation to remove blood stasis.

Objective: To investigate the effects of Exendin-4 (Ex-4) on the expressions of lipid metabolism related genes in the human liver cancer HepG₂ cells with insulin resistance (IR), and to elucidate the effect of Ex-4 in improvement of IR. Methods: The HepG₂ cells in logarithmic growth phase were induced into IR model with high concentration of insulin, then divided into control group (HepG₂ cells), IR group (HepG₂ cells were treated with insulin,HepG₂-IR cells), and Ex-4 group (HepG₂-IR cells were treated with Ex-4). Glucose oxidase (GOD-POD) kit was used to detect the consumption of glucose.The cell morphology and intracellular lipid drip formation were observed by Oil red O staining. The triglyceride (TG) level in cells was detected by kit; qRT-PCR was used to detect the mRNA expression levels of acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), sterol regulatory element-binding protein-1c (SREBP-1c) and apolipoprotein B100 (apoB100). Results: Compared with control group(HepG₂ cells), the glucose consumption in the HepG₂-IR cells in IR group was significantly decreased (P<0.01). Compared with IR group, the glucose consumption in the HepG₂-IR cells in Ex-4 group was increased (P<0.05). The Oil O red staining results showed that compared with control group, the fat percentage in the HepG₂-IR cells in IR group was increased(P<0.05); compared with IR group, the fat percentage in Ex-4 group was decreased (P<0.05). Compared with control group, the level of TG in the cells in IR group was significantly increased (P<0.01); compared with IR group, the level of TG in the cells in Ex-4 group was significantly decreased (P<0.05).The qT-PCR results showed that compared with control group, the expression levels of ACC FAS and SREBP-1c mRNA in the cells in IR group were increased (P<0.01), and the expression level of apoB100 mRNA was decreased (P<0.05); compared with IR group, the expression levels of ACC, FAS and SREBP-1c mRNA in the cells in Ex-4 group were decreased (P<0.05),and the expression level of apoB100 mRNA was increased (P<0.01). Conclusion: Ex-4 can regulate the expressions of lipid metabolism related genes in the HepG₂ cells and improve IR.

Objective: To study the expressions of cell polarity related proteins CDC42 and PAR3 during tooth germ development in the mice, and to discuss their possible roles during tooth development in the mice. Methods: The whole heads were obtained from the mouse embryo on the days 13.5,14.5,16.5 and 18.5 (E13.5, E14.5, E16.5 and E18.5) and the mice on the postnatal days 1 (PN1) and 5(PN5). The tissues were fixed in paraformaldehyde, decalcified, dehydrated, embedded in paraffin, and sectioned.The histology of tooth germ was observed by HE staining. The expressions of CDC42 and PAR3 during tooth germ development were detected by immunohistochemistry staining. Results: The HE staining results showed that E13.5, E14.5, E16.5 and E18.5 were the bud stage,the cap stage,the early and the late bell stage of tooth germ development, respectively;the tooth germ of PN1 mice showed the matured odontoblasts and ameloblasts;the tooth germ of PN5 mice showed the completed tooth crown development. The immunohistochemistry staining results showed that CDC42 expressed in the tooth germ of the mice at E13.5, E14.5 and E16.5;the CDC42 expression at E 18.5 was reduced compared with E13.5,E14.5 and E16.5; CDC42 mainly expressed in the odontoblasts and ameloblasts of the tooth germ of the mice at PN1 and PN5; PAR3 weakly expressed in the tooth germ of the mice at E13.5 and E14.5,and it was increased at E16.5 and E18.5. At PN1 and PN5, the expressions of PAR3 were decreased compared with E18.5. Conclusion: CDC42 and PAR3 participat in the mouse tooth development; during the early stage of tooth germ development, they may be involved in the proliferation and migration of mouse dental germ; during the late stage,CD42 and PAR3 may be involved in the differentiation of the odontoblasts and the ameloblasts, especially in the establishment and maintenance of cell polarity.

Objective: To compare the water absorption values, solubility values and monomer release amounts of six kinds of light curing composite resins and explore the influencing factors, and to elucidate their relationships with the material compositions and the immersing solvent. Methods: The water absorption and solubility values of six kinds of composite resins, including FiltekTM Z250(Z250), FiltekTM Z350 XT(Z350), Aelite LS Posterior (ALS), CLEARFIL MAJESTY Posterior(CMP), Neofil Nano(NN), and Tetric N-Ceram (TNC), were measured according to ISO 4049-2009. The concentrations of eluted Bis-GMA and TEGDMA were tested by liquid chromatography/mass spectrometry after the specimens were immersed in 75% ethanol solution for different time (24h, 7 d, 1 month, and 3 months). Results: The water absorption values of six kinds of composite resins were in ascending order of ALS < CMP < TNC ≈Z250 < NN < Z350,there were significant differences between groups(P<0.05);the solubility values were Z350 < NN≈ALS≈CMP≈Z250 P<0.05). The release amounts of Bis-GMA were increased slowly with the prolongation of time (P<0.05), while the values of ALS and CMP at 3 months were decreased (P<0.05); the descending order of Bis-GMA release amounts at 3 months was NN > TNC > Z250 > Z350 > ALS > CMP (P<0.05). The released TEGDMA was not found in NN and TNC. The release amounts of TEGDMA of the other four resins were increased slowly with the prolongation of time (P<0.05), and the release amount of CMP at 3 months was decreased (P<0.05); the order of amount at 3 months was ALS > CMP > Z250 > Z350 (P<0.05). Conclusion: The water absorption values, solubility values and monomer release amounts of different composite resins are different, and they are related to their organic matrix compositions and the immersing solvent.

Objective: To treat the hepatocellular carcinoma stem cells silenced by Chk-1 withcaffeine combined with 4 Gy X-ray irradiaition,and to explore their synergistic killing effects on the hepatocellular carcinoma stem cells by detecting the cell proliferation,cell cycle and apoptosis. Methods: The lentivirus silencing Chk-1 was transfected into the 293T cells;after the HepG₂ cells were infected by the lentivirus,the Chk-1 protein expression was detected by Western blotting to confirm the silencing efficency,and non-target control was established,and HepG₂-Chk-1 and HepG₂-control were named.The cells highly expressed CD133 were cultured by suspension culture method,S-HepG₂-Chk-1 and S-HepG₂-control were named respectively.After the cells were treated by caffeine,they were irradiated by 4 Gy X-ray;the proliferation activity was measured by MTT,and the cell cycle distribution and the apoptotic rate were measured by PI single staining and AnnexinⅤ-FITC double staining using FCM,respectively.For proliferation,cell cycle and apoptosis assays,there were control,caffeine,4 Gy and caffeine+4 Gy groups. Results: The Western blotting results showed that after the HepG₂ cells were infected by lentivirus,the Chk-1 protein expression was significantly decreased,but it was not obvious in non-target control group,it demonstrated that the cell models HepG₂-Chk-1 and HepG₂-control were obtained successfully.After the HepG₂-Chk-1 and HepG₂-control cells were cultivated by suspension,the CD133 protein expression were increased,it demonstrated that there were high proportion of CD133⁺ cells,they were hepatocellular carcinoma stem cells.Compared with control group,the proliferation activities in caffeine group and 4 Gy group were significantly decreased (P<0.05 or P<0.01),the percentages of cells at G₂/M phage and the apoptotic rates were significantly increased (P<0.05 or P<0.01),and the percentage of cells at S phage in caffeine group was significantly increased (P<0.05);the percentage of cells at G₀/G₁ phage in 4 Gy group was increased (P<0.05 or P<0.01),the effect was more stronger in caffeine+4 Gy group.Compared with S-HepG₂-control,the proliferation activities of S-HepG₂-Chk-1 cells in caffeine group and 4 Gy group were decreased,the apoptotic rates were increased (P<0.05 or P<0.01),and the percentage of cells at G₁/M phage was significantly decreased (P<0.05 or P<0.01). Conclusion: The hepatoccellular carcinoma stem cells silenced by Chk-1 with positive CD133 are obtained successfully; Caffeine combined with X-ray irradiation can inhibit the cell proliferation and induce the apoptosis,and enhance the G₂/M arrest,and both of them have synergistic effects.

Objective: To explore the inhibitory effect of Tum-5 gene on the tumor growth and its influence in the angiogenesis of hepatocellular carcinoma (HCC),and to illustrate its mechanism involved in anti-tumorigenesis. Methods: The human HepG₂ cells were selected in in vitro experiment and treated with different multiplicity of infection(MOI) (0,1,5,10,25 and 50) of pLXSN and pLXSN-Tum-5 virus particles for 72 h. The proliferation rates of cells in various groups were detected by MTT method.In vivo,the H22 tumor-bearing mouse models were established.The mice were divided into saline group,pLXSN group and pLXSN-Tum-5 group (n=5).The mice in saline group were intratumorally injected with normal saline,and the mice in pLXSN group and pLXSN-Tum-5 group were intratumorally injected with virus particles (MOI=5),5 times every other day.The volumes and weights of transplanted tumor and the weights of mice in various groups were measured.The CD31 expressions in transplanted tumor tissue were detected by immunohistochemical method and the microvessel density (MVD) was calculated. Results: Compared with pLXSN group,the proliferation rates of cells in pLXSN-Tum-5 group after infected with different MOI of virus particles were not significantly different (P> 0.05).The volumes and weights of transplanted tumor of the mice in pLXSN-Tum-5 group were significantly smaller than those in pLXSN group and saline group after intratumoral injection (P< 0.05 or P< 0.01).The immunohistochemical results showed that there was irregular angiogenesis in each group. Compared with saline group and pLXSN group,the mean value of MVD of the transplanted tumor tissue of the mice in pLXSN-Tum-5 group was significantly decreased (P< 0.05). Conclusion: Tum-5 can exert its antitumor activity by inhibiting the formation of neovascularization in HCC.

Objective: To study the expression of glioma-associated oncogene 1 (Gli1) in gastric carcinoma tissue,and to explore the effects of Gli1 on the proliferation and migration abilities of gastric carcinoma cells. Methods: A total of 95 cases of human gastric carcinoma tissue and paracancerous tissue were collected.Immunohistochemistry was used to detect the expression levels of Gli1 protein in gastric carcinoma tissue and paracancerous tissue; Real-time PCR method was used to detect the expression levels of Gli1 mRNA in gastric carcinoma tissue and paracancerous tissue.The human gastric cancer cell lines MKN28,BGC823,SGC7901 and immortalized gastric epithelial cells GES-1 were cultured,GES-1 as a reference,and the expression levels of Gli1 mRNA in cell lines were detected by RT-qPCR.The Gli1-siRNA and con-siRNA were transfected into the gastric carcinoma cell line BGC823,and control group,con-siRNA and Gil1-siRNA group were set up.The expression levels of Gli1 mRNA in the cells in various groups were detected by RT-qPCR; CCK8 was used to detect the proliferation of cells; Transwell migration assay was used to detect the cell migration ability.Western blotting was used to detect the expression levels of the P27,matrix metalloproteinase-2 (MMP-2),and MMP-9 proteins in the cells in various groups. Results: The positive expression rates of Gli1 mRNA and protein in gastric carcinoma tissue were significantly higher than those in paracancerous tissue (t=27.606,P<0.01; χ²=54.782,P<0.01). There were significantly differences in the expression levels of Gli1 mRNA between GES-1,MKN28,SGC7901,and BGC823 cell lines (F=86.341,P<0.01). The expression level of Gli1 mRNA in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups(F=48.322,P<0.01). The proliferation rate of gastric carcinoma cells in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups(F=54.428,P<0.01).The results of Transwell migration assay showed that the number of migration cells in Gli1-siRNA group was significantly lower than those in control and con-siRNA groups(F=257.788,P<0.01). The Western blotting results showed that the expression levels of P27,MMP-2,and MMP-9 proteins in Gli1-siRNA group had no significant differences compared with control group (P>0.05).Compared with con-siRNA group,the expression level of P27 in the cells in Gli1-siRNA group was significantly increased(t=-3.776,P=0.020), and the expression levels of MMP-2 and MMP-9 proteins were significantly dereased(P=3.497,P=0.025;t=5.487,P=0.005). Conclusion: The expression level of Gli1 in gastric carcinoma tissue is higher than that in paracancerous tissue,and the inhibition of the expression of Gli1 gene could inhibit the cell proliferation and migration.

Objective: To explore the therapeutic effect of cistanche deserticola ethanol extraction (CDE) in the rats with osteoporosis induced by ovariectomy,and to clarify its mechanism. Methods: A total of 72 female SD rats were randomly divided into control,model,estradiol (0.3 g·kg^-1),low, middle and high doses(0.5,1.0 and 2.0 mg·kg^-1)of CDE groups,and 12 rats in each group.The rat ovaries were removed by operation in the other groups to establish the osteoporosis models,while the same size of fat of the rats in control group was excised.5 d later,the rats in estradiol group were hypodermicly injected with estradiol,and the rats in CDE groups were orally administrated with corresponding doses of CDE.The same volume of diluted water was given to the rats in control and model groups.After 20 weeks,the density of right femur of the rat was detected with the method of ratio of weight and size; the mechanics index of left femur of the rat was analyzed by bone granulometer; the levels of serum alkaline phosphatase (ALP)and osteocalcin were measured by ELISA; the levels of serum Ca²⁺ and P were detected by automatic biochemical analyzer; the morphology of uterus tissue was observed by HE staining. Results: Compared with model group,the densities of right thighbone of the rats in estradiol group and low,middle and high doses of CDE groups were significantly increased (P<0.05 or P<0.01); there were no differences between low,middle and high doses of CDE groups (P>0.05).Compared with model group and low dose of CDE group,the maximum bending forces,the maximun strains,and the bone rigid coefficients in middle and high doses of CDE groups were remarkably increased (P<0.05),while the levels of ALP were decreased (P<0.05).Compared with model group,and low and middle doses of CDE groups,the level of osteocalcin in high dose of CDE group was significantly decreased(P<0.05).Compared with model group,the levels of Ca²⁺ in serum of the rats in low, middle and high doses of CDE groups were significantly increased (P<0.05); there were no significant differences between low,middle and high doses of CDE groups (P>0.05).Compared with model group,the morphology of uterus tissue of the rats in low,middle and high doses of CDE groups was improved at different degrees. Conclusion: CDE has therapeutic effect on osteoporosis through increasing the levels of serum ALP,osteocalcin and Ca²⁺ of the rats.

Objective: To detect the changes of the proliferation,cell cycle and apotopsis of liver cancer HepG₂ cells after treated with Xiaoaiping(XAP) combined with X-raysradiation,and to clarify their anti-tumor mechanisms. Methods: The human liver cancer HepG₂ cells at logarithmic phase were selected,and divided into control,XAP,2 Gy radiation and XAP + 2 Gy radiation groups.The HepG₂ cells were treated with 75 mg·L^-1 final concentration of XAP,and they were irradiated with 2 Gy X-rays 12 h later.The cell proliferation activity was detected with CCK8 kits,the cell cycle and apoptotic rates were measured by FCM with PI single staining and Annexin Ⅴ-FITC double staining,and the caspase-3 protein expression was measured by Western blotting method. Results: After the cells were treated with XAP,the proliferation activities in XAP,2 Gy radiation and XAP+2 Gy radiation groups were significantly decreased compared with control group at 12,24 and 48 h (P<0.05 or P<0.01); the percentages of cells at S and G₂/M phases in XAP,2 Gy radiation and XAP + 2 Gy radiatin groups were significantly increased compared with control group (P<0.05 or P<0.01); the apoptotic rates were significantly increased compared with control group (P<0.05 or P<0.01),especially in XAP + 2 Gy radiation group.The Western blotting results showed that caspase-3 was cleaved into 17 000 and 19 000 fragments,and its expression was also increased compared with control group. Conclusion: XAP combined with X-rays radiation could inhibit the proliferation,and induce the G₂/M arrest and the apoptosis of liver cancer HepG₂ cells; the combination of them shows a synergistic anti-tumor effect.

Objective: To investigate the antianxiety effect of schisandrin B (SchB) in the ICR mice,and to elucidate its related mechanisms. Methods: The ICR mice were divided into blank control group (administered with distilled water intragastrically), 2.5 mg·kg^-1 SchB group,5 mg·kg^-1 SchB group,and 10·mg·kg^-1 SchB group (administered with SchB intragastrically).After the successive administration for 7 d,cross maze test and zero maze test were conducted in all the mice,and then the peripheral blood and brain tissue samples of the mice in blank control group and 10 mg·kg^-1 SchB group were taken.The levels of γ-aminobutyric acid (GABA) and glutamic acid (Glu) in the peripheral blood and brain tissue of the mice were detected by ELISA,and the ratio of GABA/Glu was calculated.The expression levels of the α1 subunit of the γ-aminobutyric acid (GABAARα1) and the γ2 subunit of the γ-aminobutyric acid (GABAARγ2) in the brain tissue of the mice were detected by Western blotting method. Results: In the elevated plus maze test,the percentages of number of open arm entries in 5.0 and 10.0 mg·kg^-1SchB groups were significantly increased compared with blank control group(P<0.05 or P=0.01),and the percentages of time of open arm entries were significantly increased (P=0.05 or P<0.01).In the elevated zero maze test,the percentages of number of open arm entries of the mice in 5.0 and 10.0 mg·kg^-1SchB groups were significantly increased compared with blank control group(P<0.01),and the percentages of number of entering open arm probe were significantly increased (P<0.01).Compared with blank control group,the GABA levels in the peripheral blood and brain tissue of the mice in 10.0 mg·kg^-1 SchB group,were significantly increased(P<0.01),the Glu levels in the peripheral blood and brain tissue were significantly reduced(P<0.01),the ratios of GABA/Glu were significantly increased(P<0.01),and the expression levels of GABAARα1 and GABAARγ2 in the brain tissue were significantly increased (P<0.01). Conclusion: SchB has an antianxiety effect in the mice,and the effect may be related to its regulating the levels of GABA and Glu in the peripheral blood and brain tissue and the expression levels of GABAA Rα1 and GABAA Rγ2 in the brain tissue.

Objective: To explore the effect of gomisin A (GA) in combination with carboplatin (CBP)on the apoptosis of ovarian cancer Skov3 cells,and to illustrate the synergistic antitumor effect of GA. Methods: The ovarian cancer Skov3 cells were treated with GA combined with CBP.MTT assay was used to detectthe inhibitory rates of proliferation and the appropriate concentrations of GA and CBP were selected according to the results.The Skov3 cells were divided into control group,GA (0.04 μmol·L^-1) group,CBP (16 mg·L^-1)group and GA(0.04 μmol·L^-1)+CBP(16 mg·L^-1) group. After 48 h treatment,the cell morphology was observed by microscope,the apoptotic rate was measured by flow cytometry, the apoptotic index(AI) was observed by TUNEL staining,the mitochondrial membrane potential was detected by JC-1 staining,and the mRNA and protein expression levels of apoptosis-related genes (Bax,caspase-3,Stat3) were determined by RT-PCR and Western blotting method. Results: The inhibitory rate of proliferation of Skov3 cells in GA + CBP group(52.1%) was significantly higher than those in GA and CBP groups(P<0.01).The concentrations of GA and CBP for the best dose ratio were 0.04 μmol·L^-1 and 16 mg·L^-1,respectively. Compared with control group,the cell refractivities were decreased,the cells retracted,and part of cells were suspended in GA group and CBP groups;there were more suspended cells and cell retraction phenomenon was more prominent in GA + CBP group.The results of TUNEL staining and flow cytometry showed that the AI and apoptotic rate of cells in GA + CBP group were significantly increased compared with control group,GA and CBP group(P<0.05 or P<0.01);the JC-1 staining results suggested that the mitochondrial membrane potential of Skov3 cells in GA + CBP group was decreased(P<0.05 or P<0.01).The RT-PCR and Western blotting results showed that the expression levels of Bax and caspase-3 mRNA and protein were up-regulated(P<0.05) and the expression levels of Bcl-2 and Stat3 mRNA and protein in GA + CBP group were down-regulated compared with control,GA and CBP groups(P<0.05). Conclusion: GA can enhance the ability of CBP to induce the apoptosis of human ovarian cancer Skov3 cells,and its mechanisms may be related to the up-regulation of the Bax and Caspase-3 expressions and the down-reguation of the Bcl-2 expression;GA can synergize the induction of CBP on apoptosis.

Objective: To obtain the conditionally replicative adenovirus with triple-targeting Smac overexpression using gene recombination technology,and to explore its effects on the apoptosis and cell cycle progression of MDA-MB-231 cells. Methods: The triple-targeting Smac overexpression vector pShuttle-Egr1-Smac-HRE-hTERT-E1A-E1Bp-E1B55K was constructed by gene recombination technology,which was recombined with the skeleton vector pAdEasy in the BJ5183 bacteria (AdEasy-1+) to obtain the conditionally replicative adenovirus CRAd.pE-Smac.After the MDA-MB-231 cells were infected with CRAd.pE-Smac,the cancer cells were mimiced into hypoxic status with chemical reagent CoCl₂,then control group,CRAd.pE-Smac group,hypoxia group and CRAd.pE-Smac+hypoxia group were set up;the cells were irradiated with 4 Gy X-rays,and each group was divided into non-irradiation group and irradiation group. The Smac protein expression was detected by Western blotting assay,the apoptotic rates and the percentages of cells at different phases were detected by flow cytometry. Results: The Western blotting results showed that the Smac protein expressions were increased after infection of CRAd.pE-Smac, hypoxia and 4 Gy irradiation,especially in CRAd.pE-Smac +hypoxia+4 Gy irradiation group.The FCM results showed that the apoptotic rates in CARd.pE-Smac,hypoxia,CARd.pE-Smac + hypoxia group were increased compared with control group (P<0.05 or P<0.01),and the apoptotic rates of cells irradiated with 4 Gy were significantly increased compared with the unirradiated cells (P<0.05 or P<0.01),especially in CRAd.pE-Smac + hypoxia + 4 Gy irradiation group; the percentages of the cells at S and G₂/M phases in irradiation groups were significantly increased (P<0.05 or P<0.01),which had the similar regularity with the apoptotic change. Conclusion: After the MDA-MB-231 cells are infected with the conditionally replicative adenovirus CRAd.pE-Smac and treated with hypoxia and irradiation,the triple-targeting Smac overexpression can be achieved,and it has the role of promoting the cancer cell apoptosis and inducing the G₂/M arrest.

Objective: To discuss the protective effects of extract from fermented buckwheat flower and leaf (EFBFL) on the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and to elucidate their possible action mechanisms from the protein expression levels of peroxisome proliferator-activated receptor γ(PPARγ) and nuclear factor-κB (NF-κB) in kidney tissue. Methods: The 8weeks old male db/db mice were selected and the littermate db/m mice were used as normal control group;the db/db mice were randomly divided into model group,low dose of EFBFL group and high dose of EFBFL group,10 mice in each group.The mice in low and high doses of EFBFL groups were intragastrically given 50 and 100 mg·kg^-1 EFBFL,and the mice in normal control group and model group were intragastrically given the equal amount of distilled water once daily for 8 weeks accordingly.The levels of fasting blood-glucose (FBG) of the mice in various groups were tested respectively before and after administration.Full automatic biochemical analyzer was used to detect the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) of the mice,and the kidney index was calculated.The morphology of kidney tissue was observed by HE staining and Masson staining,and immunohistochemical method and Western blotting method were used to detect the expression levels of PPARγ and NF-κB protein in kidney tissue. Results: Compared with model group,the levels of FBG and the levels of serum SCr and BUN in EFBFL groups were significantly decreased(P<0.05 or P<0.01),and the kidney indexes were increased(P<0.01).Compared with normal control group,the kidney glomerular of the mice in model group was weaked,glomerular basement membrane was diffusely thickened,the foot process was coalesced or disappeared,and the kidney tissue fibrosis was serious; compared with the model group,the above performance of the mice in EFBFL groups were improved in different degrees,especially in high dose of EFBFL group. Compared with model group,the expression levels of PPARγ in kidney tissue of the mice in EFBFL groups were increased(P<0.01) and the expression levels of NF-κB in kidney tissue of the mice were decreased(P<0.01). Conclusion: EFBFL can improve the kidney injury in the spontaneously obese type Ⅱ diabetic db/db mice,and its possible mechanism may be related to lowering the level of blood glucose,up-regulating the expression of PPARγ and down-regulating the expression of NF-κB in the kidney tissue of the mice.

Objective: To study the effect of vascular endothelial growth factor 165b (VEGF165b) on the biological characteristics of human hepatocellular carcinoma HepG₂ cells,and to explore its mechanism. Methods: The HepG₂ cells were divided into blank group (treated with transfection reagent),control group(transfected with pcDNA3.0 expression vector)and pcDNA-VEGF165b group(transfected with pcNDA-VEGF165b).MTT assay was used to detect the survival rate of HepG₂ cells;RT-PCR and Western blotting method were used to detect the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG₂ cells;Transwell chamber assay was used to measure the migration ability of HepG₂ cells. Results: Compared with blank group,there was no significant changes in the expression levels of VEGF165b and VEGF165 mRNA and protein in the HepG₂ cells in control group(P>0.05).Compared with blank group,the expression levels of VEGF165b mRNA and protein in the HepG₂ cells in PcDNA-VEGF165b group were significantly increased (P<0.05),and the expression levels of VEGF165 mRNA and protein were significantly decreased(P<0.05).There was no significant difference in the survival rates between blank group and control group (P>0.05).The survival rate of HepG₂ cells in PcDNA-VEGF165b group was lower than those in blank group and control group,but the differences were not statistically significant (P>0.05).The cell migration experiment results showed that there was no significant difference in the migration rate between blank group and control group (P<0.05).Compared with blank group and control group,the migration rate of HepG₂ cells in PcDNA-VEGF165b group was significantly reduced(P<0.05). Conclusion: VEGF165b can inhibit the expressions of VEGF165 mRNA and protein,and VEGF165b has no effect on the proliferation of hepatocellular carcinoma cells; but it can reduce the migration of hepatocellular carcinoma cells.

Objective: To study the effect of miR-200c on the migration and proliferation abilities of breast cancer MDA-MB-231 and BT-549 cells,and to clarify the mechanism of miR-200c in inhibiting the epithelial-mesenchymal transiton (EMT) of triple negative breast cancer. Methods: The human triple negative breast cancer cell lines (MDA-MB-231 and BT-549) were chosen in this study.The cells were transiently transfected with miR-200c mimics and Lipo2000(experimental group),miR-200c negative control and Lipo2000(negative control group),and Lipo2000 alone (reagent control group); at the same time, blank control group was set up.The expression levels of vimentin and β-catenin mRNA and protein were detected by RT-PCR and Western blotting method.The proliferation rates and migration abilities of MDA-MB-231 cells and BT-549 cells after transfection of miR-200c were analyzed by CCK8 assay and wound healing assay. Results: The RT-PCR and Western blotting showed that the expression levels of vimentin and β-catenin mRNA and proteins in experimental group were decreased,and the differences were statistically significant compared with blank control group, negative control group and reagent control group(P<0.05).The CCK8 results showed that the proliferation rates of the cells in experimental group were lower than those in negative control group and reagent control group(P<0.05).The wound healing assay results showed that the recovery rate of scratch width in experimental group was lower than those in negative control group and reagent control group (P<0.05). Conclusion: miR-200c might inhibit EMT in triple negative breast cancer by regulating the expressions of β-catenin and vimentin mRNA and proteins in MDA-MB-231 and BT-549 cells and decreasing the abilities of migration and proliferation of triple negative breast cancer cells.

Objective: To investigate the protective effect of Angelica polysaccharide on the glaucoma-induced retinal neuronal injury,and to elucidate its mechanism. Methods: Eighty-four SD rats were randomly divided into control group,model group,positive drug group,low,middle and high doses of Angelica polysaccharide groups. The model of rat glaucoma was established by sclera sclerotherapy.The intraocular pressures (IOP) of rats in each group were measured.The superoxide dismutase (SOD) activities,malondialdehyde (MDA) contents and nitric oxide (NO) levels in retina tissue of the rats were measured by colorimetric assay.The expression levels of Caspase-3 mRNA and protein in retina tissue of the rats were observed by RT-PCR and Western blotting method. Results: Compared with control group,the IOP of the rats in model group at each time point were significantly increased (P<0.05); compared with model group,the IOP of the rats in different doses of Angelia polysaccharide groups were significantly decreased (P<0.05);compared with low dose of Angelica polysaccharide group,the IOP of the rats in high dose of Angelica polysaccharide group was significant decteased(P<0.05).The levels of MDA and NO in the retina tissue of the rats in model group were higher than those in control group (P<0.05),but the SOD activities were decreased (P<0.05); compared with model group,the levels of MDA and NO in retinal tissue of the rats in different doses of Angelica polysaccharide group were significantly decreased (P<0.05),and the SOD activities were significantly increased (P<0.05);compared with low dose of Angelica polysaccharide group,the above indexed had significant differences in high dose of Angelica polysaccharide group(P<0.05).Compared with model group,the expression levels of Caspase-3 mRNA and protein in different doses of Angelica polysaccharide groups were significantly decreased (P<0.05);compared with low dose of Angelica polysaccharide group,the expression levels of Caspase-3 mRNA and protein were significantly decreased(P<0.05). Conclusion: Angelica polysaccharide has protective effect on the retinal tissue of glaucoma model rats,which can decrease the levels of MDA and NO in retina tissue and increase the activity of SOD and decrease the expression levels of Caspase-3 mRNA and protein in retina tissue.It may be one of the mechanisms of polysaccharide protecting in retinal tissue nerve cells.

Objective: To study the effects of ganoderma lucidum A on the proliferation and apoptosis of human inflammatory breast cancer SUM149 cells in vitro,and to clarify their mechanisms. Methods: The human inflammatory breast cancer SUM149 cells were divided into control group and different concentrations (0.1,0.5,1.0,5.0 and 10.0 μmol·L^-1) of ganoderma lucidum A groups; four holes were set up,and the samples were taken at 24,48 and 72 h after culture.MTT assay was used to detect the inhibitory rate of proliferation of SUM149 cells.Flow cytometry was used to detect the apoptotic rate and cell cycle of SUM149 cells.The expression levels of Ki67 and Livin proteins in SUM149 cells were detected by immunocytochemical staining. Results: Compared with 0.1 μmol·L^-1 ganoderma lucidum A group,the inhibitory rates of proliferation of SUM149 cells in other concentrations of ganoderma lucidum A groups were significantly increased detectde by MTT method (P<0.05 or P<0.01) in a concentration-and time-dependent manner.The flow cytometry results showed that the apoptotic rates of SUM149 cells in different conventrations of ganoderma lucidum A groups were significantly higher than that in control group (P<0.05 or P<0.01) in a concentration-and time-dependent manner;at the same time,the cell cycle changed significantly.With the increase of ganoderma lucidum A of the concentration of ganoderma lucidum A,the pencentages of cells at G₁ phase in different concentrations of ganoderma lucidum A groups were increased(P<0.05),and the percentages of cells at S phase were decreased compared with control group(P<0.05 or P<0.01).The expression levels of Ki67 and Livin proteins in different concentrations of ganoderma lucidum A groups were decreased compared with control group (P<0.05). Conclusion: Ganoderma lucidum A can inhibit the proliferation of human inflammatory breast cancer SUM149 cells through induction of apoptosis.

Objective: To observe the development of dentinal microcracks after root canal preparation with there kinds of reciprocating nickel-titaninm instruments using an in situ cadaver model by means of micro-computed tomography (Micro CT),and to provide the reference for their clinical application. Methods: A total of 15 mandible bone specimens having at least the anterior teeth (n=60)with single root were selected and there was no dentina microcrack, then they were randomly divided into 4 groups (n=15) according to the preparation protocol:Wave One,Reciproc,One File and ProTaper Universal groups,and ProTaper Universal was used as control group.The root canals were prepared up to 25# instruments in Wave One,Reciproc and One File groups and F2 instruments in ProTaper Universal groups. After the preparation procedures, the specimens were scanned by Micro CT again,and the number of dentinal microcracks were investigated and analyzed statistically. Results: Compared with ProTaper Universal group,the incidence rates in Wave One, Reciproc and One File groups were significantly reduced(P<0.05).There were no statistical differences in the incidence rates of dentinal microcracks between Wave One, Reciproc and One File groups(P>0.05). Conclusion: Root canal preparation can cause the dentinal microcracks,so it is important to choose the types of instruments.Root canals prepared with Reciproc,Wave One and One File produce less dentinal microcracks,which can decrease the risk of vertical root fracture.

Objective: To investigate the relationships between the levels of serum interleukin-17(IL-17),interleukin-33(IL-33) and the concentration of fractional exhaled nitric oxide (FeNO) in the patients with chronic cough,and to further evaluate their effects in the pathogenesis of chronic cough. Methods: A total of 160 patients diagnosed with chronic cough for more than 8 weeks were chosenand used as chronic cough group.At the same time,60 healthy controlsreceived physical examination were selected as healthy control group.The levels of serum IL-17 and IL-33 of the subjects were examined and pulmonary function test,FeNO concentration test and comprehensive allergen test were performed.All data were analyzed by GraphPad Prism statistical software.The levels of serum IL-17 and IL-33 were compared between chronic cough group and healthy control groups.The correlations between the levels of serum IL-17 or IL-33 and lung function or FeNO in the patients with chronic cough were further analyzed. Results: The serum IL-17 and IL-33 levels of the patients in chronic cough groups were higher than those in healthy control group (P<0.05).The serum IL-17 and IL-33 levels in the patients with chronic cough showed a significant negative correlations with the percentage of the first second force expiratory volume to the estimated value(FEV1%) of the patients(r=-0.624 5,r=-0.672 2),and the level of serum IL-33 in the patients with chronic cough was positively correlated with the concentration of FeNO;the higher the concentration of FeNO,the higher the level of serum IL-33 (r_s=0.758,P<0.05).The FeNO concentration geometric mean of the patients with the serum total IgE<100 IU·mL^-1 (33 ppb) was significantly lower than those with the serum total IgE>200 IU·mL^-1 (78 ppb) and IgE 100-200 IU·mL^-1 (69 ppb) among the chronic cough patients (P<0.01),but there was no significant difference between the later two groups(P=0.082 4). Conclusion: The serum IL-17 and IL-33 may play an important role in the pathogenesis of chronic cough as proinflammatory factors.The levels of serum IL-17 or IL-33 have negative correlations with the pulmonary ventilation function in the patients with chronic cough.The increasing of serum IL-33 level may predict the formation of eosinophilic airway inflammation and reflect the severity of eosinophilic airway inflammation.

Objective: To observe the effect of distraction before discectomy(DBD) in the distraction of the height of anterior cervical space, and to explore its feasibility in restricting the over distraction. Methods: A total of 31 patients with cervical spondylotic myelopathy were treated with anterior cervical discectomy and fusion(ACDF). During surgery, the intervertebral space was distracted before discectomy, the procedure was defined as DBD technique. Before surgery,the distance from the arch top of inferior endplate of upper vertebrae of the index level to the midpoint of superior endplate of lower vertebrae was measured(H₀).The same method was used to measure the adjacent proximal and distal intervertebral space heights(H_p and H_d), and the mean value of Hp and H_d,H, was regarded as a referential height that the index intervertebral space should be restored. During operation, the intervertebral space heights before(H₁) and after(H₂) discectomy with application of DBD were measured with the aforementioned method. The pre-and post-operation index segment heights AB and A'B' were measured respectively. The change of index segment height was defined as ΔH(A'B'-AB),the index intervertebral space height after operation was defined as H₃(H₀+ΔH). The patients were divided into neck pain group and non-neck pain group according to their post-operative neck pain VAS scores.The radiographic data of the patients in two groups was analyzed and compared with statistical methods. Results: The post-operative neck pain incidence rate was 19.35%. During operation, after DBD, the intervertebral space height change ΔH₁ was (1.19±0.51)mm. In non-neck pain group,the difference between H₁ (6.95 mm±0.84 mm) and H(6.98 mm±0.70 mm) was not significant(P=0.80), and the difference between H₂ (7.31 mm±0.90 mm) and H₁(6.95 mm±0.84 mm) was significant(P<0.01).In neck pain group, the difference between H₂ (8.33 mm±1.39 mm) and H₁ (7.87 mm±1.35 mm) was significant(P<0.01).In neck pain group,the ΔH was (3.04±0.42)mm;in non-neck pain group,the ΔH was (1.70±0.51)mm;the difference between two groups was significant(P<0.01). In non-neck pain group, H₁,H₂ and H₃ had good consistency with H. Conclusion: DBD can effectively control the distraction height of the index intervertebral space within 2 mm, also convenient to let the index intervertebral space be similar with the adjacent segment and easy to follow.

Objective: To investigate the clinical efficacy of creatine phosphate sodium combined with ribavirin in the treatment of infantile viral myocarditis,and to elucidate its mechanism of the effects on myocardial enzyme levels in the children. Methods: A total of 96 children with viral myocarditis were selected; according to the random number grouping method,they were divided into observation group and control group (n=48).The myocardial enzymes and cardiac troponin I(cTnI) of the patients in two groups were detected,and anti-infection,supplementation of electrolytes,and nutritional support for myocardial treatment were performed; then intravenous infusion therapy of ribavirin was used.On this basis,the patients in observation group were given intravenous infusion of creatine phosphate sodium for 14 d.After treatment,the total effective rate of treatment of the patients in two groups,the levels of myocardial enzymes and cTnI and electrocardiogram were compared before and after treatment. Results: The total effective rate of the patients in observation group was 87.50% (42/48),and it was 70.83% (34/48) in control group;there was significant difference (χ²=4.04,P=0.04).After treatment,the levels of lactate dehydrogenase (LDH),hydroxybutyrate dehydrogenase (HBDH),aspartate transaminase (AST),creatine phosphokinase (CPK),creatine kinase isoenzyme-MB (CK-MB) and CTnI of the patients in two groups after treatment were lower than before treatment (P<0.05).The indicators mentioned above of patients in observation group were lower than those in control group(P<0.05).The total improvement rate of electrocardiogram in the observation group (89.58%) was significantly higher than that in control group (72.92%),and the difference was statistically significant (χ²=4.38,P=0.04). Conclusion: The total effective rate of creatine phosphate sodium combined with ribavirin in the treatment of infantile viral myocarditis is higher,and they can significantly reduce the levels of myocardial enzymes and improve the cardiac function; it is worth to apply in the clinical treatment.

Objective: To analyze the association of the genetic variations of rs2383206 and rs2383207 in 9p21 region with the coronary heart disease (CHD) in the Chinese Han population,and to explore whether chromosome 9p21 is a susceptibility region for CHD. Methods: Case-control study was conducted.A total of 580 CHD patients were selected as case group,and 539 cases of non-cardiovascular disease patients or normal people with matched age and sex were selected as control group.The rs2383206 and rs2383207 loci of the subjects were genotyped with Sequenom MassARRAY time of flight mass spectrometer (TOF). Results: The smoking,waist-to-hip ratio (WHR),hypertension,diabetes mellitus,systolic blood pressure (SBP),diastolic blood pressure (DBP) and total cholesterol (TC) of the subjects in two groups were statistically different (P<0.05).Compared with control group,the ratios of patients with smoking,hypertension and diabetes mellitus of the patients in case group were increased (P<0.05); the WHR,SBP,DBP and TC level were also increased (P<0.05).There was no significant difference in the genotypic distribution of rs2383206 between case group and control group (χ²=4.623,P>0.05),while the genotypic distribution of rs2383207 was statistically different (χ²=8.936,P<0.05);the distribution frequency of AA genotype in case group (8.3%) was significantly lower than that in control group (13.6%)(P<0.05). Conclusion: Smoking,WHR,hypertension,diabetes mellitus,SBP,DBP and TC may be the risk factors for CHD; the AA genotype of 9p21 rs2383207 loci may be the protective genotype of CHD.

Objective: To explore the expression of CC chemokine receptor 6 (CCR6) in human adenoid cystic carcinoma tissue of salivary glands,and to clarify the mechanism of CCR6 in the nerve invasion of human adenoid cystic carcinoma. Methods: Immumohistochemical method(SP) was performed to detect the expression of CCR6 in 30 cases of cancer tissue of the patients with adenoid cystic carcinoma (including 18 cases in nerve invasion group and 12 cases in non-nerve invasion group) and 22 cases of normal salivary gland tissue. The relationship between the positive expression rate of CCR6 and the clinicopathological features of the patients were analyzed. Results: CCR6 was positively expressed in 22 cases among 30 cases of salivary adenoid cystic carcinoma tissue,and the positive expression rate was 73.33%.The immunohistochemical staining showed that CCR6 mainly expressed in the cell membrane and cytoplasm of cancer cells. All of the 22 cases of normal salivary gland tissue were negative to CCR6 expression;the difference of the positive expression rate of CCR6 between two groups was statistically significant(P<0.05). The positive expression rate in nerve invasion group and non-nerve invasion group were 94.44%(17/18) and 41.67%(5/12),respectively; there was statistically significant between two groups(P<0.05). The positive rates of CCR6 in the patients with salivary gland adenoid cystic carcinoma with different ages,sex and pathological types had no statistically significant differences(P>0.05). Conclusion: CCR6 plays an important role in the development of human salivary adenoid cystic carcinoma and is closely related to the nerve invasion.

Objective: To compare the clinical efficacy and safety between etanercept and infliximab in the treatment of adolescent ankylosing spondylitis (AS),and to provide reference for guiding clinical medication. Methods: Sixty male adolescents with AS from our Hospital were used as the subjects and randomly divided into etanercept group and infliximab group (n=30).All patients in the 24 weeks before treatment and during the treatment were not taken the disease-modifying anti-rheumatic drugs (DMARDS).The pillow-wall distance,finger-ground distance,Sch ber test,AS activity index (BASDIA),AS functional Index (BASFI),C-reactive protein (CRP) levels,erythrocyte sedimentation rate (ESR),MRI performance and other indicators of the patients in two groups were measured and evaluated 12 and 24 weeks after treatment,and statistical analysis was performed to assess the clinical efficacies of the two drugs and the adverse effects were recorded. Results: Etanercept and infliximab had good effects on the AS 24 weeks after treatment,and the pain of low back and lumbosacral of the patients in two groups were significantly reduced or disappeared,and the total clinical efficacies in two groups were 96% and 93%, respectively (P>0.05).There were significant improvements in the indexes of morning stiffness,finger-ground distance,pillow-wall distance,Sch ber test,BASDAI scores,BASFI scores,ESR and CRP levels of the patients in two groups 12 and 24 weeks after treatment compared with before treatment(P<0.05).During treatment,1 case of erythema was found in etanercept group;2 cases of moderate infection,2 cases of digestive system reaction, and 1 case of pruritus were found in infliximab group;the differences in the incidence rates of adverse effects between two groups were statistically significant (P<0.05). Conclusion: Etanercept and infliximab have good therapeutic effects in the adolescent patients with AS.They can improve the joint symptoms of hip joint involved in AS;the short-term efficacies of the patients in two groups have no significant difference,but the incidence rate of adverse effects of etanercept is slightly less than that of infliximab.

Objective: To repaire the posterior tooth defects in the elderly people with CEREC chairside CAD/CAM restorations, and to evaluate the clinical curative effect 1 year after treatment. Methods: A total of 52 posterior teeth from 48 aged patients were selected. Among them, 24 cases were onlays, 18 cases were inlays, and 10 cases were crowns. The inlays/onlays/crowns were designed and manufactured by CEREC chairside CAD/CAM system, and then bonded with resin cement. After 12 months, the patients were followed up. The teeth with restorations were assessed by using the modified USPHS criteria including restorations, tooth, and periodontal. At the same time, the patient's satisfaction was evaluated. Results: In all of the 52 restorations, the success rates of inlay, onlay and crown were 94.4%, 91.7% and 80.0%, respectively; among them, one inlay was fractured, one onlay did not close the edge, one onlay's edge integrity was mild defect, one teeth was fractured, one crown's adjacent relationship was not close, and one crown appeared papillitis. Conclusion: The restorations manufactured by CEREC chairside CAD/CAM system have a good short-term clinical curative effect. It is an effective way to repair the posterior tooth defects in the elderly people.

Objective: To explore the influence of gallnut extract as a root canal irrigants in the curative effect of chronic periapical periodontitis,and to clarify its mechanism. Methods: A total of 192 patients diagnosed with chronic periapical periodontitis were selected and randomly divided into control group (distilled water),hydrogen peroxide group,sodium hypochlorite group and gallnut extract group (n=48).In the process of root canal treatment,four irrigants were used respectively,then the curative effects 1 week,6 months,12 months after treatment were observed and compared. Results: After root filling treatment,the cold and heat stimulation of the tooth was insensitive in clinical examination,percussion(-),loose degree ≤ Ⅰ°,no swollen gums,bleeding on probing(-),no fistula,normal function of the exercise of chewing,decreased or disappeared transmission area in periapical bone in X-ray photo,and those metioned above were considered effective treatment.After 6 months of treatment,the effective rate in gallnut group was 89.4%,and there were no statistically significant differences compared with hydrogen peroxide group (91.1%) and sodium hypochlorite group(91.5%) (P>0.05),but there was significant difference compared with control group (67.4%) (P<0.05).After root filling treatment for 12 months,the effective rate in gallnut group was 83.7% and there were no statistically significant differences compared with hydrogen peroxide group(83.3%) and sodium hypochlorite group(87.8%) (P>0.05),but the ratio of tooth from improvement to recovery in gallnut group was higher than those in hydrogen peroxide group and sodium hypochlorite group. Conclusion: Gallnut extract as a root canal irrigants has good antibacterial and cleaning effect,and can achieve the aim of curing the chronic root periapical inflammation and preventing the infection.

Objective: To investigate the clinical features,diagnosis and treatments of primary breast diffuse large B-cell lymphoma(PB-DLBCL),and to provide the basis for its clinical treatmemt. Methods: Three patients with PB-DLBCL were examined by imaging,pathology and bone marrow cytology. The curative effect was observed,while the related literatures were also reviewed. Results: Three patients with PB-DLBCL were female and had the clinical manifestations (painless breast mass) and their imaging findings were similar to breast cancer. the 3 patients,one patient underwent excisional biopsy in the assessment of breast nodules,followed by chemotherapy with 4 cycles of R-CHOP +4 cycles of CHOP+ 1 prophylactic intrathecal injection treatment,no recurrence was found in 1 year's follow-up;the other 2 patients were diagnosed by core needle biopsy,receiving 2 cycles of CHOP and 1 cycle of CHOP, respectively,getting very good partial remission (VGPR) and complete remission (CR);they still received chemotherapy,and the process was smooth. Conclusion: Core needle biospy along with immunohistochemical method combined with clinical data is an effective technique for the diagnosis in the patients with when unilateral painless mass as the first symptom and highly suspected as PBL.The elementary role of chemotherapy of R-CHOP/CHOP is preferred for the patients.Central nervous system (CNS) prophylaxis may improve the prognosis in the early treatment.

Objective: To discuss the therapeutic effect of related periodontal surgical treatment and aesthetic re-repair in the patient with residual root and crown of anterior teeth induced by unfitting prosthesis and to clarify the curative effect of periodontal surgical treatment and aesthetic re-repair in the anterior teeth area,and to provide guidance for making the program of clinical treatment. Methods: Two years after repair of the anterior teeth of a patient,the restorations loosed;on the one hand the patient had a toothache of chewing food accompanying gingival bleeding,on the other hand the upper labial frenum was too high,furthermore the upper left incisor apical area showed low density images.After the consultation of periodontics,endodontics,prosthodontics and other disciplines doctors, anterior teeth root canal re-treatment and periodontal basic treatment were adopt as well as crown lengthening combined with labial freneetomy to restore the normal biological width,additionally apply not only guided bone regeneration(GBR) but also platelet rich fibrin(PRF) to repair the defects of apical bone.One month later,provisional restorations were put into use and then three months later permanent repairs were conducted. Results: The corresponding gingival and gingival papilla shape recoverd perfectly and the height of gingiva coordinated with that of adjacent teeth,in the meantime the upper labial frenum attached normally and there was not obvious space between upper incisors;the morphology and function examination of restoration in maxillary anterior region were both well.The follow-up X-ray showed that the bone density of the defect area was the same as that of the peripheral bone without obvious boundaries. Conclusion: The aesthetic repair of thin biotype referring to anterior area,especially in the complicated re-treatment cases,the intervention of the appropriate periodontal surgical treatment is not only beneficial to obtain the obvious short-term effect,but also positive to stabilize long-term effects.

Objective: To investigate the diagnosis and treatment of one patient with pneumocephalus caused by congenital mastoid dysplasia,and to clarify the clinical features,diagnostic methods and treatment strategies of intracranial accumulation of pneumocephalus. Methods: The patient with ineffective right upper limb activity accompanied stupid speech for 12 h was admitted to hospital. After admission, the head CT and MRI examination were performed again,and the patient was diagnosed as pneumocephalus.The paitent scheduled for stoma repair, neurotrophic treatment,infection prevention and other symptomatic treatments were performed after operation;the patient was instructed avoid cough forcefully,blowing nose,defecating and sneezing to increase the intracranial pressure. Results: Due to congenital dysplasia of mastoid wall, local thinning bones and intense swimming choking cough of the patient destroyed the intracranial pressure balance to form pneumocephalus,the patient scheduled for stoma and damaged dura repair;when discharged from hospital, the patient's right upper limb muscle strength and language function returned to normal;the head CT results showed that pneumocephalus disappeared completely. Conclusion: Pneumocephalus is common in clinic,and its reason is diversiform; it should be combined with the patient's history and imaging findings to explore the causes,the most reasonable treatment measures should be performed in order to relieve the patient's symptoms of increased intracranial pressure,and promote the recovery of neural function.

Objective: To observe the effect of bilateral thoracoscopic surgery for invasive thymoma complicated with myasthenia gravis, and to explore feasibility of surgical procedure. Methods: Bilateral thoracoscopic extended thymectomy was performed in one patient with invasive thymoma complicated with myasthenia gravis,with complete resection of thymus and anterior mediastinal adipose tissue through bilateral approach,and the curative effect was observed. Results: Bilateral thoracoscopic thymectomy for invasive thymoma had radical effects with rapid recovery after surgery,avoiding large surgical trauma and more complications in open surgery and flawed in removing contralateral fat and ectopic thymus with unilateral thoracoscope. Conclusion: Bilateral thoracoscopic extended thymectomy has radical effects for invasive thymoma complicated with myasthenia gravis,confirming its feasible surgical procedure.

Objective: To investigate the clinical characteristics of patient with giant ovarian mature cystic teratoma with contralateral ovarian endometriosis cysts, and to analyze the diagnostic experience and treatment process. Methods: The chinical materials of one case of giant hepatic mature cystic teratoma with contralateral ovarian endometriosis cyst was analyzed retrospectively, and the related literatures were reviewed. Results: The patient was admitted to the hospital for peritoneal giant mass in a physical examination. A large abdominal cysts was seen by gynecologic ultrasound. In the left ovary, a 61 mm×48 mm cystic light mass was seen, and the viscous liquid was found in it.The abdominal CT showed a huge lump of mixed density in the right abdomen. The areas of density of fat, calcification density and density of soft tissue were found in the abdomen. The liver was pressured by the mass.The right ovarian tumor size was about 40cm×35cm×30cm, up to the diaphragm, down to the pelvic tumor;the posterior wall of tumor adhered the left lobe of liver and the liver was squeezed to the upper right.The size of the left ovarian endometriosis cyst was about 7cm×6cm×6cm. The operation was to remove the right ovarian tumor and left ovarian endometriosis cyst, and to reconstruct bilateral ovaries. The pathological diagnosis was right ovarian mature cystic teratoma and left ovarian endometriosis cyst. After 2 years of follow-up, no recurrence of the tumor was found.The patient was conceived with in vitro fertili zation frozen embryo transfer(IVF-ET). Conclusion: The huge mature teratoma combined with contralateral ovarian endometrial cyst, which adhereds to the liver, is rare. Surgical treatment will gain good prognosis. IVF-ET should be early done when the patient is going to be infertile after surgery.

Objective: To explore the correct test ways of the ductus venosus(DV) blood flow spectrum of the normal fetal and the rule of changes of the normal fetal DV blood flow spectrum along with gestational week,and to establish the reference ranges of the parameters of blood flow. Methods: A total of 320 cases of single fetal pregnant women within 11-40 weeks were selected as the objects.Using color doppler echocardiography,the ventricular systolic peak velocity(SV),atrial systolic trough(aV),pulse index(PI) and resistance index(RI) were measured,and the ratio of ventricular systolic peak velocity trough ratio(S/a) from ultrasonic standard section was calculated.All of these were analysed statistically. Results: The doppler spectrum of normal fetal DV blood flow showed the same three-phase waves,SV was increased with the increase of gestational age(F=27.00,P=0.000),and aV was elevated with the increase of gestational age(F=389.81,P=0.000),while PI(F=65.41,P=0.000),RI(F=58.82,P=0.000) and S/a ratio(F=47.79,P=0.000) were decreased with the increase of gestational age. Conclusion: The performance of color Doppler flow imaging of normal fetal DV showed that the blood flow peak speed is increased along with the increase of gestational age and PI,RI and S/a are reduced with the increase of gestational age,and RI has the largest decline,which has important diagnosis value.