Objective: To investigate the changes of ultrastructure of myocardial cells and the expression levels of tumor necrosis factor α(TNF-α) in myocardium tissue of the diabetic rabbits during early myocardial ischemia and their mechanisms,and to provide the theoretical basis for studying the pathogenesis of diabetic myocardial ischemia injury. Methods: A total of 42 healthy male New Zealand white rabbits were chosen.The method of intravenous injection of alloxan in ear vein was used to establish the diabetic models in 26 rabbits among 42 rabbits; the method of partial ligation of left anterior descending coronary artery by thoracotomy was used to establish the diabetic myocardial ischemia models in 22 model rabbits with diabetes.Atotal of 16 (modeling succesfully) were divided into diabetic myocardial ischemia 7 d group and 28 d group;other 16 rabbits were used as sham operation groups at the relevant time points(7 and 28 d),and there were 8 rabbits in each group.The ultrastructure of myocardial cells was observed by transmission electron microscope; the apoptosis index (AI) of myocardial cells in ischemic myocardium tissue of the rabbits was detected by TUNEL method; the expression levels of TNF-α in ischemic myocardium tissue of the rabbits were detected by real-time RT-PCR method. Results: In sham operation groups,the nuclei of myocardial cells was bigger,the nuclear chromatin was in uniform distribution,the myofibril ordered arrangement,the sarcomere was neat;the mitochondria between myofibril were rich,and their structures were clear.For the rabbits in diabetic myocardial ischemia group,their early-stage cardiocyte nuclei were irregular in shape,myofibrils was loose in structure,the part of myofibril was broken,and the mitochondrial hyperplasia between the myocardial cells was obvious.Compared with sham operation groups,the AI of myocardial cells and the expression levels of TNF-α in the ischemic myocardial cells of the rabbits in diabetic myocardial ischemia 7 d and 28 d groups were significantly increased(P<0.05),and the indexes mentioned above in diabetic myocardial ischemia 7 d group was higher than those in diabetic myocardial ischemia 28 d group(P<0.05). Conclusion: The destruction of ultrasturctue and obvious apoptosis of the myocardial cells of the rabbits occur in the early stage of diabetic myocardial ischemia.The increasing of expression level of TNF-α may involve in the myocardial ischemic injury.

Objective: To investigate the effects of telmisartan on the expressions of activin A and its receptors in non-infarction area of left ventricle in the heart failure rats after myocardial infarction,and to clarify the mechanism of improvement effect of telmisartan on myocardial collagen deposition. Methods: A total of 27 rats were divided into sham operationgroup(n=9), model group(n=20) and telmistartan group(n=10).The left anterior descending coronary arteries of the rats in model group and telmisartan group were ligated,and the left anterior descending coronary arteries of the rats in sham operation group were not ligated.After 5 weeks,there were 6 surviving rats in each group.The rats in sham operation group were treated with 0.5%CMC(10 mL·kg^-1),the rats in model group were treated with 0.5%CMC (10 mL·kg^-1),and the rats in telmisartan group were treated with telmisartan (30 mg·kg^-1). All the rats were treated with medicines by intragastric administration for 4 weeks.The whole cardiac hypertrophy index and left ventricular hypertrophy index of the rats were measured;the mRNA expression levels of activin A,ActRⅡA,ActRⅡB, ColⅠ,and ColⅢ in the non-infarction area of the rats were detected by RT-PCR and the ratio of ColⅠ/ColⅢ was calculated;the expression intensities of activin A protein in the myocardium tissue in non-infarction area of the rats in various groups were observed by immunohistochemical staining. Results: Compared with sham operation group,the whole cardiac hypertrophy index and left ventricular hypertrophy index of the rats in model group were significantly increased (P<0.01);the mRNA expression levels of activin A,ActRⅡA and ActRⅡB,ColⅠ,ColⅢ and the ratio of ColⅠ/ColⅢ were also increased (P<0.01).The immunohistochemical staining results showed that the expression level of activin A protein in noninfarction area of left ventricle of the rats in sham operation group was lower,and the expression level of activin A protein in non-infarction area of left ventricle of the rats in model group was higher. Conclusion: Telmisartan may improve the myocardial collagen deposition by inhibiting the expressions of activin A and its receptors during heart failure.

Objective: To investigate the effect of human growth hormone releasing hormone receptor splice variant type 1 (GHRHR SV1) on the proliferation of human liver cancer HepG2 cells, and to clarify the proliferation effect of GHRHR SV1 on the human cancer cells. Methods: The GHRHR SV1 plasmids were transfected into the human HepG2 cells to construct the HepG2-SV1 cell line. HepG2 group(HepG2 cells),HepG2-empty group(HepG2-pcDNA3.0 cell line) and HepG2-SV1 group(HepG2-SV1 cells) were set up. PCR and Western blotting methods were used to identify the HepG2-SV1 cell line;CCK-8 method was used to detect prolifernation rate of cells;colony formation assay was used to detect the colony formation rate of cells;cell wound healing assay was used to evaluate the migration rate of cells. Results: The PCR and Western blotting results showed the HepG2-SV1 cell line expressed GHRHR SV1 steadily.The CCK-8 results showed that the proliferation rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The colony formation assay results showed that the colony formation rate of HepG2-SV1 cells in HepG2-SV1 group was 3.5 times higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The cell wound scratch assay results showed that the migration rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05). Conclusion: GHRHR SV1 could increase the proliferation of HepG2 cells.

Objective: To explore the effect of sub-inhibitory concentration of imipenem on the bio-activities of methicillin resistant Staphylococcus aureus(MRSA)and illuminate the inhibitory effects of carbapenem antibioty on the activity of MRSA and their mechanisms,and to provide the basis for using the carbapenem antibiotics in the treatment of MRSA infection. Methods: Five ST239 type of MRSA clinical isolates were selected and were co-cultured with 1/10 and 1/2 minimal inhibitory concentration (MIC) of imipenem for 1.5,6.0 and 12.0 h,which were divided into control group(no imipenem),1/10MIC group(1/10MIC of imipenem),and 1/2MIC group(1/2MIC of imipenem).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes fibronectin A(fnbA),staphylococcal protein A(spa),α-hemolysin(Hla),leukocidin D(lek-D),leukocidin E(lek-E), and enterotoxin A in various groups;Spectrophotometry was used to detect the proliferation activity of MRSA strains in various groups in vitro. Results: After co-culture for 6.0 and 12.0 h,the proliferation activities of 5 trains of MRSA in 1/2MIC group in vitro were significantly decreased compared with control group (P<0.01).The relative mRNA expression levels of 6 virulence-related genes of 5 strains of MRSA in 1/10MIC and 1/2MIC groups were significantly decreased compared with control group(P<0.01).After co-culture for 12.0 h,the mRNA expressions of all the tested virulence-related genes were not found. Conclusion: The sub-inhibitory concentration of imipenem shows obviously inhibitory effect on the mRNA expressions of multiple virulence-related genes of ST 239 type of MRSA strains,and higher concentration of imipenem can suppress the proliferation of MRSA strains in vitro.Imipenem shows the potential value in the treatment of the severe MRSA infected patients.

Objective: To study the inhibitory effect of salidroside on the proliferation of fibroblast-like synoviocytes with rheumatoid arthritis in human(HFLS-RA) induced by tomor necrossi factor-α(TNF-α),and to clarify the molecular mechanism of its control effect on rheumatoid arthritis(RA). Methods: The HFLS-RA were cultured in vitro,then treated with TNF-α and different concentrations of salidroside.The cells were divided into normal control group(0 μg·L^-1TNF-α),model control group(10.0 μg·L^-1TNF-α)and 12.5,25.0,50.0,and 100.0 μmol·L^-1 salidroside groups(10.0 μg·L^-1TNF-α+salidroside).The proliferation activity was detected by MTT mehthod;the expression levels of β-catenin,matrix metalloproteinase-7(MMP-7),and Cyclin-D1 in supernatant of the cells were detected by ELISA method;the expression level of β-catenin protein in cells was detected by Western blotting method. Results: Compared with normal control group,the proliferation activity of the HFLS-RA in model control group was significantly increased (P<0.05);compared with model control group,the proliferation activities of the HFLS-RA in 12.5 and 25.0 μmol·L^-1 salidroside groups were decreased but there were no significant differences(P>0.05),and the proliferation activities of the HFLS-RA in 50.0 and 100.0 μmol·L^-1 salidroside groups were significantly decreased (P<0.05).Compared with normal control group,the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in model control group were increased(P<0.05).Compared with model control group,the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in 12.5 and 25.0 μmol·L^-1 salidroside groups were decreased,but there were no significant differences(P>0.05);the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in 50.0 and 100.0 μmol·L^-1 salidroside groups were decreased(P<0.05).The Western blotting results showed that the expression level of β-catenin protein in the cell in model control group was higher than that in normal control group(P<0.01);the expression levels of β-catenin protein in the cells in 12.5 and 25.0 μmol·L^-1 salidroside groups were lower than that in model control group,but there were no significant differences(P>0.05);the expression levels of β-catenin protein in the cells in 50.0 and 100.0 μmol·L^-1 salidroside groups were lower than that in model control group(P<0.05). Conclusion: Salidroside could inhibit the proliferation of HFLS-RA,and its control effect might be related to the regulation of Wnt/β-catenin single pathway.

Objective: To investigate the effect of hypoxia on the expression of forkhead box P3 (FOXP3) in human oral squamous cell carcinoma (OSCC) cells, and to clarify its possible epigenetic mechanism. Methods: Two kinds of OSCC cell lines, FaDu and OECM-1, were cultured under normoxic or hypoxic conditions for 18 h. The relative expression levels of FOXP3 mRNA and protein in the cells were detected by Real-time RT-PCR and Western blotting method. The histone modification levels on the FOXP3 gene promoter, including acetylation of histone 3 lysine 4 (H3K4ac), trimethylation of histone 3 lysine 4 (H3K4me3) and lysine 27 (H3K27me3), were analyzed by Chromatin Immunocipitation (ChIP) and quantitative PCR (ChIP-qPCR). The relative expression levels of histone deacetylase 3 (HDAC3) mRNA and inhibitory rates of FOXP3 mRNA expression in the HDAC3-knockdown FaDu cells were investigated by Real-time qPCR and ChIP-qPCR. Results: Compared with normoxic condition, the relative expression levels of FOXP3 mRNA in FaDu and OECM-1 cells under hypoxic condition were decreased by 65.6% and 75.7% (P<0.01). The Western blotting results indicated that compared with normoxic condition, the expression levels of FOXP3 protein in FaDu and OECM-1 cells under hypoxic condition were decreased. The ChIP experiment results showed that compared with normoxic condition, the levels of H3K4ac and H3K4me3 on FOXP3 gene promoter in FaDu cells were decreased under hypoxic condition (P<0.01), while the H3K27me3 level was not changed.In HDAC3-knockdown FaDu cells, compared with control cells, the inhibitory rates of the expressions of H3K4ac and H3K4me3 on FOXP3 gene promoter under hypoxia condition were decreased (P<0.05), so did expressions the FOXP3 mRNA expression (P<0.05). Conclusion: Hypoxia could suppress the expression of FOXP3 by HDAC3-mediated down-regulation of H3K4ac on FOXP3 gene promoter in the human OSCC cells.

Objective: To investigate the effect of Superfine Cordyceps militarisv Powder on the immune functions of the mice,and to provide basis for improving the utilizable ratio of Cordyceps militaris. Methods: A total of 40 mice were randomly divided into negative control group,low dose (0.166 5 g·kg^-1),middle dose (0.333 3 g·kg^-1),and high dose (0.999 9 g·kg^-1) of Superfine Cordyceps militaris Powder groups,and there were 10 mice in each group. The mice in different doses of Superfine Cordyceps militaris Powder groups were administered with the Superfine Cordyceps militaris Powder for 30 d by intragastric infusion respectively,whereas,the mice in negative control group were administered with the same volume of water for 30 d by intragastric infusion.The body weights,the spleen indexes and thymus indexes of the mice in various groups were measured; the lymphocyte transformation abilities of the mice in various groups were observed by lymphocyte transformation experiment;the levels of delayed-type hypersensitivity reaction of the mice were examined with toe incrassation; the number of antibody forming cells,phagocytic rate and phagocytic index of chicken erythrocytes phagocytized by peritoneal macrophages of the mice were detected. Results: Compared with negative control group,the body weights,the spleen indexes and thymus indexes of the mice in different doses of Superfine Cordyceps militaris Powder groups had no significant differences (P>0.05).The lymphocyte transformation abilities of the mice in middle and high doses of Superfine Cordyceps militarisPowder groups were higher than that in negative control group(P<0.05). The levels of delayed-type hypersensitivity reaction of the mice in middle and high doses of Superfine Cordyceps militaris Powder groups were higher than those in low dose of Superfine Cordyceps militaris powder group and negative control group(P<0.05).The numbers of antibody forming cells of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05).The phagocytic rates and phagocytic indexes of chicken erythrocytes phagocytized by peritoneal macrophage of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05). Conclusion: Superfine Cordyceps militaris Powder can strengthen the immune functions of the mice,and the recommended doses are 0.333 3 and 0.999 9 g·kg^-1.

Objective: To study the effects of compound schisandra extracts (CSE) (schisandra,astragalus,acanthopanax,and rhodiola)on the exhaustive swimming time and the levels of blood urea nitrogen(BUN),serum lactic acid(LD),liver glycogen and muscle glycogene,superoxide dismutase(SOD)activity,and malonaldehyde(MDA) level in the mice and to charify its anti-fatigue effect and the mechanism. Methods: Eighty male ICR mice were randomly divided into blank control group, 50 mg·kg^-1 CSE group,100 mg·kg^-1 CSE group,and 200 mg·kg^-1CSE group;there were 20 mice in each group. The mice were administered orally for 30 d.Then 10 mice were randomly selected for exhaustive swimming test in each group and the exhaustive swimming time of the mice was recorded.The remaining 10 mice in each group were used for 90 min swimming,then all the mice were sacrificed and the blood and tissue samples were taken for the measurement of the levels of BUN,LD,liver glycogen and muscle glycogen,the SOD activity and MDA level;the total inhibitory rate of oxidation of CSE in vitro was determined by linoleic acid-ferric thiocyanate method. Results: Compared with blank control group,the exhaustive swimming time of the mice in 50,100,and 200 mg·kg^-1 CSE groups were significantly increased (P<0.01); the levels of BUN and LD of the mice in 100 and 200 mg·kg^-1 CSE groups were significantly decreased (P<0.05 or P<0.01);the levels of liver glycogen and muscle glycogen of the mice in 100 and 200 mg·kg^-1 groups were significantly increased(P<0.05 or P<0.01); whereas the SOD activities were significantly increased and the levels of MDA were decreased(P<0.05 or P<0.01).Compared with 100 mg·kg^-1 CSE group,the levels of serum BUN and LD of the mice in 200 mg·kg^-1 CSE group were decreased (P<0.01), and the levels of liver glycogen and muscle glycogen were increased(P<0.05).The total inhibitory rate of oxidation of 5 g·L^-1CSE was 76.94%. Conclusion: CSE has an anti-fatigue effect and the mechanism may be related to anti-oxidation effect.

Objective: To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein. Methods: A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured; the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry. Results: Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P<0.05 or P<0.01);the indexes metioned above of the mice in MUC1-MBP+ R848 were higher than those in MUC1-MBP+BCG group;the IL-4 levels had no significant difference(P>0.05).Compared with control group,the proprotions of CD3⁺T,CD4⁺T,and CD8⁺ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01). Conclusion: Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.

Objective: To investigate the effect of the extract of Schisandra chinensis on the matrix metalloproteinases(MMPs) system in kidney tissue of the diabetic rats,and to explore its protective effect on the kidney tissue from the matrix degradation perspective. Methods: STZ was used to establish rat models of diabetes mellitus. A total of 45 diabetic rats were randomly divided into model group,extract of Schisandra chinensis group and Benazepril group,and there were 15 rats in each group.Another 15 rats were selected and used as normal control group.12 weeks after administration,the routine blood and urine biochemical indexes,the histological changes,blood glucose (BG), blood urea nitrogen(BUN),serum creatinine(Scr),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterin(LDL-C),total cholesterol(T-CHO),and triglyceride(TG) levels, excretion rates of albuminuria and proteinuria of the rats in various groups were detected; the expression amounts of fibronectin (FN),type Ⅳ collagen (Col Ⅳ),and matrix metalloproteinase inhibitor metalloproteinase-2 (TIMP-2) in kidney tissue of the rats were detected by immunohistochemical method; the activity of matrix metalloproteinase-2 (MMP-2) was detected by zymography. Results: Compared with model group,the glomeruli matrix accumulation of the rats in extract of Schisandra chinensis group was significantly improved,the excretion rate of albuminuria,LDL-C level and serum MDA level were decreased(P<0.05),the activities of CAT(P<0.01)and SOD(P<0.05)in kidney tissue were increased,and the level of MDA in kidney tissue was decreased(P<0.05). The immunohistochemistry results showed that compared with model group,the expression amounts of FN,Col Ⅳ,and TIMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group were significantly decreased.The zymography results showed that compared with model group,the activity of MMP-2 in kidney tissue of the rats in extract of Schisandra chinensis group was significantly increased(P<0.05). Conclusion: Extract of Schisandra chinensis has protective effect on the kidney tissue of the diabetic rats induced by STZ,and the mechanism may be related to the inhibition of oxidative stress and the improvement of MMP-2 activity as well as the inhibition of TIMP-2 expression which could improve the matrix degradation.

Objective: To explore the effects of voriconazole on the proliferation and morphology of Acanthamoeba cultivated in vitro,and to clarify the killing effects of voriconazole against the trophozoites and cysts of Acanthamoeba. Methods: The Acanthamoeba polyphaga at logarithmic phase were selected and divided into control group and experiment groups(2.5 and 25.0 mg·L^-1).The Acanthamoeba in each group was collected at 24,48,72, and 96 h after drug administration,respectively.Then the concentrations of Acanthamoeba were calculated and the proliferation curves were drawn;inverted microscope was used to observe the morphology,activity and adherence of Acanthamoeba;the ultrastructures of Acanthamoeba were observed under electron microscope. Results: Compared with control group,the numbers of Acanthamoeba polyphaga in experiment groups were significantly decreased(P<0.01).The morphology of Acanthamoeba changed significantly under inverted microscope,and the shape of Acanthamoeba transformed from the trophozoites with irregular spiny filopodia to circular cysts.Even a large number of cell debris was observed.Different degrees of damage and even necrosis of Acanthamoeba in experiment groups were found under electron microscope. Conclusion: Certain concentration of voriconazole can effectively inhibit the proliferation of Acanthamoeba and change the morphology and ultrastructure and kill the trophozoites and cysts of Acanthamoeba cultivated in vitro.

Objective: To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism. Methods: The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells; the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method; the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits. Results: The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation, the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation, the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation. Conclusion: ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

Objective: To prepare the velvet antler polypeptide-collagen/chitosan composite materials, and to investigate its promotive effect on cicatrization of mandibular defect and possible mechanism. Methods: The collagen and chitosan solution were mixed.The composite material was prepared by glutaraldehyde crosslinking method.The microstructure of the composite material was observed by transmission electron microscope (SEM).The unilateral mandibular defect models of 36 rabbits were established.The rabbits were divided into experiment and control groups,and each group was divided into 4-,8-and 12-week subgroups, and there were 6 rabbits in each sub group.The rabbits in experiment group were implanted with velvet antler polypeptide-collagen/chitosan composite materials and the rabbits in control group were treated.4,8 and 12 weeks after operation,the histology of bone defect and peripheral nerve reconstruction of the rabbit models were detected by CT; the expression of vascular endothelial growth factor (VEGF) in bone tissue of the rabbits was detected by immunohistochemistry;the ultrastructure of bone defect was observed by SEM. Results: The structure of composite materials had layered folds and the inner diameter of the stent became larger and mainly dominated by sheet structure,which was the ideal structure of biological materials.4 weeks after operation,the new bone was formatted in experiment group,most of the new bone like-tissue materials were degraded,and the VEGF expression showed an increasing trend; 8 weeks after operation,the trabecular bone in the bone defect of the rabbits in experiment group was increased obviously and the expression of VEGF was decreased.12 weeks after operation,the new bone formation and the density in experiment group was consistent with the normal tissue,and the expression level of VEGF returned to normal.At each the point after operation,the degree of bone defect healing and bone formation rate in experiment group were obviously prior to control group. Conclusion: Velvet antler polypeptide-collagen/chitosan composite material has the promotive effect on the fracture healing of mandibular defect of the rabbits and its possible mechanism may be related to promoting the expression of VEGF.

Objective: To explore the influence of total alkaloids of Corydalis Ochotensis(TAOCO) on the behavior and pathomorphology of brain tissue of the rats with Alzheimer's disease(AD) induced by β-amyloid protein 25-35(Aβ_25-35),and to clarify its therapeutic effects on the AD rats. Methods: The Wistar rats were divided into mormal control group(treated with 0.5 mL·100 g^-1 distilled water)(n=9),model group(treated with 0.5 mL·100 g^-1 distilled water)(n=9),positive drug group(treated with 1.75 mg·kg^-1 donepezil hydrochloride)(n=9), and low,middle and high doses (treated with 2.0,4.0 and 8.0 mg·kg^-1) of TAOCO groups(n=8,n=9,n=9).The rat AD models were made by injecting Aβ_25-35 into hippocampus.On the 14th day after operation,the rats were administered for 7 d.Morris water maze test was used to detect the spatial learning and memory ability of the rats;dark avoidance task was used to observe the passive avoidance ability of the rats; the pathomorphology of the cerebral cortex and hippocampus of the rats were detected. Results: The Morris water maze test results showed that compared with model group,the latency to platform of the rats in low dose of TAOCO group was decreased on the 4th and 5th days(P<0.05); the latency and swimming distance to reach the platform of the rats in middle dose of TAOCO group were decreased on the 5th day(P<0.05),and the starting angle of the rats was reduced on the 3th day(P<0.05); the latencies to reach the platform of the rats in high dose of TAOCO group were decreased on the 2nd and 5th days(P<0.05),and the starting angle of the rats was decreased on the 6th day(P<0.01). Compared with model group,on the 7th day,the time of staying on platform,distance of staying on platform,time of staying on platform/total time,distance of staying on platform/total distance of the rats in different doses of TAOCO groups were all increased significantly(P<0.05) within 1.5 min.Compared with model group,the times of crossing platform and time of staying in effective area of the rats in low and high doses of TAOCO groups were significantly increased(P<0.05).The dark avoidance task results showed that compared with model group,the error latencies and the error times of the rats in different doses of TAOCO groups had no significant differences on the 2nd day(P>0.05).Compared with model group,there was no obvious improvement of the cerebral cortex and hippocampus injury of the rats in low and middle doses of TAOCO groups.In high dose of TAOCO group,the cerebral cortex and hippocampus injury of the rats were significantly improved. Conclusion: TAOCO can improve the learning and memory function of the AD rats and reduce the pathological injury of brain tissue of AD rats.

Objective: To observe the effects of Shen Hong Bu Xue Granule(SHBXG) on the expression levels of erythropoietin(EPO)mRNA in kidney tissue and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA in bone marrow tissue of the mice with blood deficiency, and to investigate the protective effect of SHBXG on the blood deficiency mice and its mechanism. Methods: The mouse model of blood deficiency was established with acetylphenylhydrazine(APH) and cytoxan(CTX).A total of 60 mice were divided into blank control group,model group,Compound E-jiao Slurry group and low,middle,high doses of SHBXG groups(n=10).The serum,kidney and bone marrow tissues from all the mice were collected after 14 d consecutive administration.The levels of red blood cells(RBC),hemoglobin level(HGB),hematocrit(HCT),leukocytes(WBC),and platelet(PLT) in blood of the mice were detected by automatic blood analyzer;the levels of serum EPO and GM-CSF of the mice in various groups mice were detected by ELISA method;the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice were detected by RT-PCR method. Results: The results of automatic blood analyzer showed that the levels of RBC,HGB,HCT,and PLT of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group (P<0.05 or P<0.01);the levels of WBC of mice in Compound E-jiao Slurry group and high dose of SHBXG group were increased significantly compared with model group(P<0.05).The ELISA results showed that the levels of serum EPO and GM-CSF of the mice in Compound E-jiao Slurry group and different doses of SHBXG groups were increased significantly compared with model group(P<0.05).The PC-PCR results showed that the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice in Compound E-jiao Slurry group and middle and high doses of SHBXG groups were increased significantly compared with model group(P<0.05 or P<0.01). Conclusion: SHBXG could improve the blood deficiency symptom in the mice with blood deficiency,and its mechanism may be related to increasing the expression levels of EPO mRNA in kidney tissue and GM-CSF mRNA in bone marrow tissue of the mice.

Objective: To study the biotransformation rule of the chemical components of total ginsenosides from ginseng stems and leaves tranformed by submerged fermentation of ganoderma lucidum, and to lay the foundation for further development and research of the medicinal fungus of ginseng stems and leaves. Methods: The total ginsenosides from ginseng stems and leaves were fermented by ganoderma lucidum,the changes of chemical components before and after fermentation were measured by thin-layer chromatography;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were detected by ultraviolet spectrophotometry;the levels of ginsenoside Rb₁,Rg₃,Rd,CK,Re,Rg, and Rh₁ before and after fermentation were measured by high performance liquid chromatography.The human hepatocellular carcinoma SMMC-7721 cells were divided into blank control group,low,medium and high doses of total ginsenosides from ginseng stems and leaves groups and medicinal fungus of ginseng stems and leaves groups.The dosages of total ginsenosides from ginseng stems and leaves and medicinal fungus of ginseng stems and leaves were 5,10 and 20 mg·L^-1,respectively.The inhibitory rate(IR) of the cells was determined by MTT assay. Results: The results of thin-layer chromatography showed that the saponins before and after fermentation occured mutual transformation;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were 749.98 and 602.26 mg·g^-1,respectively;the levels of panaxadiol saponins Rb₁,Rg₃,Rd and CK before fermentation were 24.54,2.21,87.22 and 0 mg·g^-1,and the levels of panaxatriol saponins Re,Rg and Rh₁ were 151.34,77.02,and 3.06 mg·g^-1.After fermentation,the level of Rb₁ was decreased by 61.45%,the levels of Rg₃,Rd and CK were increased by 238.91%,34.43% and 268.00%.The levels of Re and Rg₁ were decreased by 63.75% and 64.41%,and the level of Rh₁ was increased by 408.88%;the IR of the SMMC-7721 cells in medium dose of medicinal fungus of ginseng stems and leaves group was higher than that in medium dose of total ginsenosides from ginseng stems and leaves group(P<0.05);the IR of the SMMC-7721 cells in high dose of medicinal fungus of ginseng stems and leaves group was higher than that in high dose of total ginsenosides from ginseng stems and leaves group(P<0.01). Conclusion: The total ginsenosides from ginseng stems and leaves transformed by the submerged fermentation of ganoderma lucidum can significantly change its chemical components,which have stronger anti-proliferation activity in the SMMC-7721 cells.

Objective: To investigate the effects of different doses and infusion methods of recombinant mouse interleukin-18(rmIL-18) on the survival time and tumor diameter of the mice with hepatocellular carcinoma, and to elucidate the rational application of rmIL-18 in vivo. Methods: A total of 60 Babl/C mice were randomly divided into 5 μg rmIL-18 intraperitoneal injection group, 0.5 μg rmIL-18 intraperitoneal injection group, 0.5 μg rmIL-18 tumor injection group, cytotoxic T lymphocyte(CTL) intraperitoneal injection group, CTL tumor injection group and saline control group;there were 10 mice in each group. From the 10th day of inoculation, the mice in different rmIL-18 groups were injected with the corresponding doses and methods. The mice in different CTL groups were injected with tumor-specific CTL (1×10⁶/mouse) by intraperitoneal and intratumoral injection. The mice in saline control group were injected with an equal volume (100 μL) of saline, the injections were performed 10 times. The diameters of mice were measured weekly and the survival time was recorded. Results: Compared with 5 μg rmIL-18 intraperitoneal injection group, 0.5 μg rmIL-18 intraperitoneal injection group and saline control group, the tumor growth rate of the mice in 0.5 μg rmIL-18 tumor injection group was decreased (P<0.01)and the survival rate of the mice was increased (P<0.01); compared with 0.5 μg rmIL-18 intraperitoneal injection group, the tumor growth rate and the survival rate of the mice in CTL intraperitoneal injection group were decreased (P<0.01);compared with 0.5 μg rmIL-18 tumor injection group,the tumor growth rate and the survival rate of the mice in CTL tumor injection group were decreased (P<0.01). Conclusion: The best way for rmIL-18 anti-tumor effect is tumor injection and the effect has a dose-dependent manner.

Objective: To establish the alcoholic fatty liver(AFL) animal models,and to explore the protective effect of Zhi Zi Da Huang Tang (ZZDHT) on the rats with AFL and its dosage. Methods: A total of 54 SD rats were divided into normal control group (n=10) and model control group (n=44).The rats in model control group were given alcohol by lavage (50% ethanol solution 6.0 mL·kg^-1) combined with high-fat feed to establish the rat models of AFL.After 4 weeks,the rats in model control group were randomly divided into model group(treated with water),simvastatin group (10 mg·kg^-1), low and high doses (5.0 and 10.0 g·kg^-1) of ZZDHT groups,and there were 10 rats in each group.Every 2 weeks during the process,the body weights and levels of serum total cholesterol (TC), triglyceride (TG), aspertate aminotransferase (AST), and alanine aminotransferase (ALT) were detected.After 4 weeks,the body weights and liver weights of the rats were detected; the levels of TC,TG,AST,ALT,alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum and liver tissue of the rats of were detected;the levels of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and interleukin-1β(IL-1β) in serum of the rats were detected; the morphology of liver tissue was observed by HE staining and the pathological examination was performed. Results: Compared with normal control group,the levels of serum TC,AST,ALT, TG,TNF-α,IL-6,and IL-1β of the rats in model group 4 weeks after administration were significantly increased (P<0.05 or P<0.01).Compared with model group, the levels of serum TC,TG,AST,and ALT of the rats in low and high doses of ZZDHT groups 4 weeks after administration were decreased (P<0.05 or P<0.01); the levels of serum TNF-α and IL-6,IL-1β of the rats in high dose of ZZDHT group were decreased (P<0.01);the levels of serum TNF-α and IL-1β of the rats in low dose of ZZDHT group were decreased (P<0.01); the levels of serum ADH and ALDH in liver tissue of the rats in low and high doses of ZZDHT groups and simvastatin group were decreased (P<0.05 or P<0.01).The HE staining result showed that compared with model group, the pathological conditions of the liver tissue of the AFL rats in ZZDHT groups were significantly improved. Conclusion: ZZDHT can significantly improve the liver injury caused by high fat diet combined with alcohol and fat liver lesions.

Objective: To investigate the promotive effect of 4,5,6,7-tetrabromobenzotriazole (TBB) on the apoptosis of the human cervical cancer HeLa cells,and to clarify its effect mechanism in related signaling pathways. Methods: The human cervical cancer HeLa cells at logarithmic growth phase were divided into control group(without TBB) and experiment group(with TBB).MTT assay was used to detect the survival rate of the HeLa cells; the morphology of HeLa cells was observed under inverted microscope; Annexin Ⅴ-FITC/PI double staining and flow cytometry(FCM) were used to determine the apoptotic rates of the HeLa cells; the expression levels of apoptosis-related proteins p-Akt,Akt,Bcl-2,Bax,cleaved-caspase-3,and Pro-caspase-3 were detected by Western blotting method. Results: The MTT results showed that the survival rates of the HeLa cells in experiment group were significantly decreased(P<0.01) in a dose-dependent manner compared with control group.The apoptotic morphology of the HeLa cells in experiment group were found as cell shrinkage and karyopyknosis under inverted microscope. The Annexin Ⅴ-FITC/PI double staining and FCM results showed that the apoptotic rates of the HeLa cells in experiment group (3,6,12 and 24 h) were higher than that in control group (P<0.05 or P<0.01). The Western blotting results showed that compared with control group,the expression levels of the anti-apoptotic proteins p-Akt and Bcl-2 in HeLa cells in experiment group were decreased obviously,whereas the expression levels of the pro-apoptotic proteins Bax and cleaved-caspase-3 were increased and the expression levels of Pro-caspase-3 were decreased. Conclusion: TBB may promote the apoptosis of human cervical cancer HeLa cells by inhibiting the Akt signaling pathway.

Objective: To observe the effect of Toudingyikezhu extract on the morphology of liver tissue and endoplasmic reticulum stress(ERS) of the rats with nonalcoholic fatty liver disease(NAFLD),and to clarify the possible mechanism of Toudingyikezhu of intervention of NAFLD. Methods: The NAFLD rat models were established by intraperitoneal injection of carbon tetrachloride solution combined with high-fat diet.A total 60 SD rats were randomly divided into control group,model group,low,middle and high doses of Toudingyikezhu extract groups,phosphatidylcholine group(n=10).The doses of Toudingyikezhu extracts were 0.50,1.00 and 2.00 g·mL^-1,and the dose of phosphatidylcholine was 0.20 g·kg^-1, 4 weeks after administration,the body weights and liver wet weights were detected,and the liver indexes were calculated.The texture,elasticity,color and morphology of liver tissue of the rats were observed by HE staning.The fatty degeneration,inflammation and necrosis scores of liver tissue of the rats were calculated.The levels of serum tumor necrosis factor -α(TNF-α),interleukin-1 (IL-1) and interleukin-6(IL-6) of the rats were detected by ELISA method. Immunohistochemical SP method was used to determine the expression levels of GRP78 protein, and PCR method was used to determine the expression levels of GRP78 mRNA. Results: Compared with control group,the morphology of liver tissue of the rats in model group were abnormal under light microscope;the liver index, fatty degeneration,inflammation and necrosis scores of liver tissue of the rats in model group were increased(P<0.05),and the expression levels of GRP78 protein and mRNA were decreased(P<0.01). Compared with model group, the liver tissue pathology of the rats had different degrees of improvement; the fatty degeneration,inflammation and necrosis scores of liver tissue and the serum TNF-α,IL-1, IL-6 levels of the rats in different doses of Toudingyikezhu extract groups were decreased(P<0.05 or P<0.01);the expression levels of GRP78 protein and mRNA were decreased (P <0.05 or P<0.01). Conclusion: Toudingyikezhu extract may inhibit or block the ERS,reduce the synthesis of lipids and accelerate its decomposition in order to resist the injury of liver cells of the NAFLD rats by down-regulating the GRP78 expression.

Objective: To explore the protective effect of ellagic acid on the acute liver injury induced by carbon tetrachloride (CCl₄) of the mice, and to explore its possible mechanism. Methods: A total of 50 mice were randomly divided into normal control group, model group, low, medium and high doses of ellagic acid groups (n=10). The mice in normal control group and model group were treated with 1% sodium carboxymethyl cellulose solvent by intragastic administration, and the mice in ellagic acid groups were treated with 160, 320,and 480 mg·kg^-1 ellagic acid by intragastic administration, respectively. After 14 d, the mice in model group and different doses of ellagic acid groups were intraperitoneally injected with 10 mL·kg^-1 0.1% CCl₄, while the mice in normal control group were intraperitoneally injected with 10 mL·kg^-1plant oil. 16 h later, all the mice were sacrificed and the body weights and the liver indexes of the mice were detected;the serum levels of alanine transaminase(ALT), aspartate transaminase(AST) were detected; the activities of superoxide dismutase(SOD) and levels of GSH-Px, malonalde hyde(MDA) and catalase(CAT) in liver tissue of the mice were detected. Results: There were no significant differences of the body weights of the mice between each group before and after treatment(P>0.05). Compared with normal control group, the liver indexes and the levels of serum ALT and AST of the mice in model group and different doses of ellagic acid groups were significantly increased (P<0.05).Compared with model group, the liver indexes of the mice in different doses of ellagic acid groups were decreased(P<0.05);the serum levels of ALT and AST of the mice in high dose of ellagic acid group were significantly decreased (P<0.05), while the CAT level in liver homogenate was significantly increased (P<0.05); the levels of GSH-Px in liver homogenate of the mice in medium and high doses of ellagic acid groups were significantly increased (P<0.05); the activities of SOD in liver homogenate of the mice in different doses of ellagic acid groups were significantly increased (P<0.05), and the MDA levels were significantly decreased (P<0.05). Conclusion: The ellagic acid has the protective effect on acute chemical liver injury in the mice induced by CCl₄ and the most effective dose is 480 mg·kg^-1;its mechanism may be related to the anti-oxidation.

Objective: To explore the effects of KISS1 gene transfected by lentivirus on the proliferation, invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms. Methods: The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected. The lentiviral vectors were builted and transfected the KISS1 gene, and the cells were divided into control group (treated with PBS), empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells; Transwell chambers method was used to detect the invasion and migration abilities of the cells. Results: Among LoVo, SW620, SW480, HCT-116, and HT29 cells, the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells, so they were chosen as the research carrier. After transfected with lentiviral vectors, the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene, and the MOI was over 80%. Compared with control group and empty vector group, the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P<0.05); the proliferation abilities of the cells in over-expression group were decreased (P<0.05); the invasion ability and migration ability of the cells in over-expression group were decreased (P<0.05). But the differences of proliferation ability,invasion ability and migration ability of the cells between control group and empty vector group were not statistically significant (P>0.05). Conclusion: The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation, invasion and migration abilities of the colorectal cancer cells, and the mechanism may be related to the expression of KISS1 protein.

Objective: To investigate the preventive and protective effects of Schisandrin B in the mice of Alzheimer's disease (AD), and to clarify its mechanism. Methods: Fifty Balb/c mice were randomly divided into blank group, model group, pasitive control group, low dose of Schisandrin B group(0.1 g·kg^-1)and high dose of Schisandrin B group(0.5 g·kg^-1);there were 10 mice in each group. Step-through test was conducted after last administration to detect the latencies and number of errors of the mice in various groups, and the brain tissue was taken. Congo red staining was to detect the morphology changes of cells and neuronal amyloidosis in brain tissue of the mice. The levels of ROS in brain tissue of the mice were tested by Flow Cytometry. The contents of MDA, the levels of LDH, and the activities of CAT, GSH-Px and SOD in brain tissue of the mice were tested by biochemical method. Western blotting method was used to detect the expression levels of signaling pathway proteins Nrf2 and Keap1 in brain tissue of the mice. Results: Compared with model group, the latencies of the mice in low and high dose of Schisandrin B groups were increased (P<0.01) and the number of errors in step-through tests was decreased (P<0.05 or P<0.01).The Congo red staining results showed that compared with model group,the neuronal amyloidosis in brain tissue of the mice in Schisandrin B groups was decreased significantly. Compared with model group, the levels of ROS, LDH and the contents of MDA in brain tissue of the mice in low and high doses of Schisandrin B groups were decreased (P<0.05 or P<0.01), and the activities of CAT, SOD and GSH-Px were increased (P<0.01). Compared with low dose of Schisandrin B group, the content of MDA and the activities of SOD and GSH-Px in brain tissue of the mice in high dose of Schisandrin B group were increased (P<0.001). Compared with model group, the expression level of Nrf2 protein in brain tissue of the mice in low dose of Schisandrin B group was increased (P<0.01), while the expression level of Nrf2 protein in brain tissue of the mice in high dose of Schisandrin B group was decreased (P<0.01); the expression levels of Keap1 protein in brain tissue of the mice in low and high doses of Schisandrin B groups was decreased (P<0.01). Conclusion: Schisandrin B could decrease the level of peroxidation in brain tissue of the mice and reduce the oxidative damage and improve the memory function of the AD mice. The mechanism is related to the activation of Nrf2 signaling pathway which improve the activity of antioxidant enzymes.

Objective: To evaluate the effects of several kinds of surface treatment methods on the bond strength of zirconia prosthesis, and to provide references for improving the bond strength of zirconia prosthesis. Methods: Sixty zirconia blocks(10 mm×10 mm×2 mm) were divided into 6 groups according to the surface treatmentmethods:sandblasting, silica coating, Z-PRIME Plus, sandblasting+Z-PRIME Plus, sandblasting+silica coating,and silica coating+Z-PRIME Plus groups.The zirconia-resin specimens were fabricated using Scotchbond^TM Universal; shear bond test was performed to detect the shear bond strength after treated with water storage(37℃) for 24 h by universal mechanical testing mechine; the fracture types were observed by stereoscopic microscope.One fracture specimen was randomly chosen from each group,and the morphology of the specimen was examined under scanning electron microscope. Results: The shear bond strength of the specimen in Z-PRIME Plus group was higher than those in the other groups(P<0.05). The adhesive failure was predominantly observed in sandblasting,silica coating,sandblasting+silica coating and silica coating+Z-PRIME Plus groups.There were 4 cases of mixed failure in Z-PRIME Plus group and there were 8 cases of mixed failure in sandblasting+Z-PRIME group.No cohesive failure was observed in all the groups.The scanning electron microscope results showed that the fracture surface occurred on the resin-zirconia interface of the specimens in sandblasting group, Z-PRIME Plus group, and sandblasting+Z-PRIME Plus group.The silica-zirconia fracture of specimens occurred in silica coating group and silica coating+Z-PRIME Plus group.The silica-resin interface fracture of specimens partially occurred in sandblasting+silica coating group. Conclusion: The application of Z-PRIME Plus can significantly improve the bond strength between zirconia and resin.

Objective: To use polylactic-co-glycolic acid(PLGA) as vector material to prepare the compound total flavonoids of Rhizoma Drynariae(TFRD) and total flavonoids of Chrysanthemum(TFC) sustained release microcapsuleTF-PLGA microcapsules,and to investigate the the best preparation technique of TF-PLGA microcapsules and their sustained release characteristics in vitro. Methods: The TF-PLGA microcapsules were prepared with TFRD,TFC, and PLGA by emulsifying-solvent evaporation technique under certain conditions.With the encapsulation efficiency(EE) as the evaluation indicator,the optimal formulation was verified by single factor experiment and orthogonal design;the general morphology,the particle size and distribution of the microcapsules were observed by light microscope(LM) and scanning electron microscope(SEM);the cumulative drug release rate of TF-PLGA microcapsules was detected by constant temperature commotion method in vitro and the release curves of the TF-PLGA were drawn. Results: The optimal prescription was as follows:the concentration of PLGA was 140 g·L^-1,oil phase volume was 1.4 mL,emulsifying speed was 900 r·min^-1,emulsifying time was 5 min,the average EE was(83.89±2.30)%,and the average drug loading rate(DL) was(5.90±0.07)%.The LM and SEM resluts showed that the TF-PLGA microcapsules presented as round ball,the average particle size was(44.34±14.68)μm, and the distribution was relatively narrow.The drug release in vitro results showed that the initial drug release rate(24 h)was about 40%,and the cumulative drug release rate was over 90% after 50 d. Conclusion: The TF-PLGA sustained release microcapslue has better drug-loaded and sustained-release effects with simple preparation technique and better repeatability.

Objective: To discuss the relationship between the geographic environment and blood urea nitrogen(BUN) reference values of the healthy people, and to explore the distributional rule of BUN reference values of the healthy people, and to provide the scientific foundation for establishing the BUN reference value standards of different areas. Methods: A total of 23 geographic factors and 33521 BUN reference values of healthy adults measured by 403 medical facilities from 23 provinces, 4 municipalities and 5 autonomous regions were collected. The spatial autocorrelation analysis was used to determine the spatial autocorrelation of the sample data; the correlation analysis was used to detect the factors which correlated significantly with the BUN reference values;the multiple linear regression, principle component analysis and ridge regression analysis were respectively used to construct the predicted models; the paired-sample t test was used to choose the optimal model; the distribution map of BUN reference values was built by geostatistic analysis. Results: There were 5 geographic factors, latitude(X₂), altitude(X₃), annual mean temperature(X₅), annual mean relative humidity(X₆) and annual precipitation(X₇), correlated significantly with the BUN reference values. The regression equation of optimal model was ?=5.112+0.000 127 1X₂+0.000 094 61X₃-0.000 140 5X₅-0.000 136 8X₆-0.000 139 1X₇±0.531 0; the distribution map of predicted values of the BUN reference values was obtained. Conclusion: The overall trend of BUN reference values is low in the east and high in the west. The BUN reference value is negatively associated with the altitude. If the geographic data of a certain region could be obtained, the BUN reference value of this region will be predicted.

Objective: To detect the neutrophil extracellular traps (NETs) formation in the peripheral blood of the patients with recurrent respiratory tract infection, and to evaluate the effect of sulbactam sodium/cefoperazone sodium on the formation of NETs. Methods: A total of 36 patients with recurrent respiratory tract infection (case group) and 30 healthy volunteers (healthy control group) were selected. The NETs formation of subjects in two groups was detected by confocal microscope and scanning electron microscope (SEM). According to the appearance of neutrophils, the formation of NETs was classified as grade Ⅰ, Ⅱ and Ⅲ,the number of NETs formation cells of subjects in two groups was calculated. The formation of NETs of the patients in case group were detected before and after treated with sulbactam sodium/cefoperazone sodium. Results: The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group was more than that in healthy control group (P<0.05); while the number of NETs formation cells of grade Ⅲ of the patients in case group was less than that in healthy control group (P<0.05). The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group were significantly decreased (P<0.05), while the number of NETs formation cells of grade Ⅲ was significantly increased (P<0.05) after treated with sulbactam sodium/cefoperazone sodium. Conclusion: A lot of NETs with high antibacterial function can be formed in the patients with recurrent lower respiratory tract infection, and sulbactam sodium/cefoperazone sodium can inhibit the formation of NETs.

Objective: To detect the interleukin-6 (IL-6) levels and expressions of applipoprotein E (ApoE) in the Alzheimer's disease (AD) patients with different body-mass indexes (BMI), and to explore the diagnotic value of ¹¹Pittsburgh comound-B (¹¹C-PIB) combined with ¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) PET/CT imaging in AD. Methods: A total of 58 AD patients were divided into four groups according to their BMI:low BMI group(BMI <18.5 kg·m^-2,n=18),normal BMI group(18.5 kg·m^-2 ≤ BMI <24.9 kg·m^-2,n=13),high BMI group(24.9 kg·m^-2 ≤ BMI <29.9 kg·m^-2,n=12)and obese group(BMI ≥ 29.9 kg·m^-2,n=15). All the patients underwent PET/CT imaging (¹¹C-PIB and ¹⁸F-FDG). The sensitivity,specificity and accuracy rate, the expression rates of ApoE (ε2, ε3, and ε4) and the levels of serum IL-6 were detected. The relationship between BMI and the expression rates of ApoE and the serum levels of IL-6 were analyzed by Spearman analysis. Results: The sensitivity, specificity, and accuracy rate of the patients in low BMI group diagnosed by ¹¹C-PIB and ¹⁸F-FDG PET/CT were 87.5%,80.0%, and 84.6%, which were higher than those diagnosed by ¹¹C-PIB (55.6%,50.0%, and 53.8%) or ¹⁸F-FDG (42.9%,50.0%, and 46.2%)alone (P<0.05). The serum levels of IL-6 and BMI of the AD patients had a negative correlation(r=-0.407,P=0.002).The expression rate of ApoE ε4 allelic gene(60.3%) of the AD patients was higher than those of ε2(18.9%) and ε3 allelic genes(20.7%), but there was no correlation between the BMI and the expression rates of different ApoE allelic genes of the AD patients(r=-0.028,P=0.833). Conclusion: ¹¹C-PIB and ¹⁸F-FDG PET/CT has a high diagnotic value in the AD patients. ¹¹C-PIB and ¹⁸F-FDG combinated with serum IL-6 level and BMI could diagnose and evaluate AD more exactly.

Objective: To investigate the application of Coronary Artery Disease-Reporting and Data System (CAD-RADS) in the diagnosis of coronary atherosclerotic heart disease(CAD) and its risk factors, and to clarify the effective strength of different risk factors in the diagnosis of CAD by using CAD-RADS. Methods: All the data of 266 patients, who were initially suspected with CAD and underwent CT angiography, were collected and diagnosed by using CAD-RADS and were divided into CAD group(n=174) and non-CAD group(n=69). The informations of age, gender, hypertension, hyperlipemia, diabetes, smoking, serum uric acid (UA) levels, and plasma fibrinogen (FIB) levels of the patients in two groups were analyzed;single factor analysis and multivariate Logstic regression analysis were used to analyze the risk factors of CAD diagnosed by CAD-RADS. Results: Compared with non-CAD group,the ratios of male, hypertension, diabetes,smoking of the patients in CAD group were increased (P<0.05),and the age and the level of UA of the patients in CAD group were also increased (P<0.05). The Logistic regression analysis results showed that age and diabetes were the independent risk factors for CAD diagnosed by CAD-RADS. Conclusion: There are many independent risk factors for CAD diagnosed by CAD-RADS, and age and diabetes are the most correlated risk factors for CAD.

Objective: To explore the necessity of the preventive use of antibiotics and the effects of age and operation time on the efficacy of inguinal hernia repair without tension, and to elucidate the clinical significance of the preventive application of prophylactic antibiotics in inguinal hernia repair without tension. Methods: A total of 228 patients with inguinal hernia repair without tension were selected, amomg them 42 cases with high infection factors were treated with antibiotics (treated group),and 186 cases were not treated with antibiotics(untreated group) during the preoperative period.The prophylactic antibiotics were given 30 min before surgery, and the conventional dose was not used more than 48 h after surgery. All the cases were treated with artificial repair materials for the procedure of inguinal hernialrepair without tension.The age, highest body temperature, white blood cell count, operation time, hospitalization time, and postoperative body temperature of all the 228 cases were recorded and analyzed statistically. Results: The preoperative and postoperative white blood cell counts had significant differences between the patients<60 years and the patients ≥ 60 years in untreated group (P<0.05);there were no significant differences in the operation time, hospitalization time and body temperature between the patients<60 years and the patients ≥ 60 years in untreated group (P>0.05).Compared with the patients with the operation time>90 min, the white blood cell count and hospitalization time of the patients with the operation time ≤ 90 min were increased (P<0.05).There were no significant differences of the body temperature and age between the patients with the operation time >90 min and the patients with the operation time ≤ 90 min (P>0.05). The white blood cell count, operation time,hospitalization time and postoperative body temperature of the patients between treated group and untreated group had no significant differences (P>0.05). Conclusion: The use of antibiotics in the high-risk patients and non-use of antibiotics in the majority of elective inguinal hernia repair without tension can ensure the safe and performability of the patients.

Objective: To investigate the feasibility,moderate-and short-term efficacies of laparoscopic surgery in the resection of the gastric gastrointestinal stromal tumors(GIST) with the tumor diameter more than 5 cm. Methods: The clinicopathological data of 26 gastric GIST patients with the diameter more than 5 cm underwent laparoscopic complete resection were retrospectively analyzed. The postoperative recovery, complications and moderate-and short-term prognosis of the patients were observed. Results: All the patients underwent laparoscopic surgery resection without open operation.According to the location and growth types of tumors,the total gastrectomy was performed in 1 case, the approximal gastrectomy was performed in 2 cases,the distal gastrectomy was performed in 2 cases,and the gastric local resection was performed in the rest 20 cases.The mean diameter of tumors was (5.94±1.28) cm,the mean operational time was (84.23±27.02) min,and the mean blood loss was (72.38±34.24) mL.The postoperative recovery time of bowel function was (2.77±0.65)d and the mean length of postoperative hospitalization time was (7.04±2.24) d.There was 1 case of intraperitoneal hemorrhage and 1 case of anastomotic stenosis;they recovered after conservative therapy.The postoperative pathology results(according to the modified NIH standard) indicated the number of moderate-risk patients was 18(69.23%),and the number of high-risk patients was 8(30.77%).A total of 12 cases accepted the adjuvant therapy with imatinib.There was 1 case of hepatic multimetastasis,and there was no local recurrence or death in the other patients. Conclusion: The moderate-and short-term efficacies of laparoscopic surgery in the gastric GIST patients with the tumor diameter more than 5 cm is safe and feasible.

Objective: To explore the feasibility and clinical efficacy of stratified suture of lateral pharyngeal wall(SSLPW) combined with soft palate radiofrequency coblation in the treatment of the patients with severe obstructive sleep apnea hypopnea syndrome(OSAHS). Methods: The clinical data of 21 severe OSAHS patients underwent lateral pharyngoplasty(LP) combined with soft palate radiofrequency coblation (LP group) and 39 severe OSAHS patients underwent SSLPW combined with soft palate radiofrequency coblation (SSLPW group) were selected.The apnea hypopnea index(AHI),lowest SaO₂(LSaO₂),Epworth sleepiness scale(ESS) scores,and related postoperative complications of the patients in two groups were analyzed before and after operation.The successful rates of operation of the patients in two were compared. Results: In LP group, 1 patient was cured (5%), 19 patients were improved markedly (90%), 1 patient was effective (5%), and no patient was invalid; the successful rate was 95.2%(20/21).In SSLPW group, 2 patients were cured (5.1%), 33 patients were improved markedly (84.6%), 4 patients were effective (10.3%), and no patients was invalid; the successful rate was 89.7%(35/39); there was no statistical difference in the successful rate of operation of the patients between two groups (P>0.05). There was statistically significant improvementof the subjective symptoms. Allthe patients returned to normal subjective swallowing functions without nasal pharyngeal reflux and dysphagia in one month follow-up after operation.Compared with before operation,the AHI and ESS scores of the patients in two groups after operation were decreased(P<0.05);the LSaO₂ scores were increased(P<0.05). Conclusion: The AHI, LSaO₂ and ESS scores are significantly improved in the OSAHS patients after treated with SSLPW combined with soft palate radiofrequency coblation. The method is a valid option for the severe OSAHS patients.

Objective: To study the influence of prophylactic bilateral salpingectomy during hysterectomy in ovarian function of the premenopausal women with benign uterine disease and its preventive effect on the pelvic diseases(malignant/benign pelvic lesions or pelvic inflammatory disease). Methods: A total of 100 patients with benign uterine disease underwent hysterectomy were collected and divided into two groups,including 50 patients underwent simultaneous hysterectomy and prophylactic bilateral salpingectomy (prevention group) and 50 patients underwent only hysterectomy (control group).The operative time,blood loss, evacuation time,and hospitalization time of the patients in two groups were detected; the antral follicle count, levels of serum estradiol(E2),follicle stimulating hormone(FSH) and luteinizing hormone(LH)) and the incidence of perimenopausal symptoms of the patients in two groups before operation and six months, 1 year after operation were compared; the incidence of pelvic disease of the patients after operation were compared. Results: The operative time,blood loss,evacuation time, and hospitalization time of the patients between two groups were not significantly different(P>0.05).Compared with before operation,the antral follicle count and the E2 levels of the patients 6 months and 1 year after operation in two groups were decreased, and the FSH and LH levels of the patients were increased (P<0.01).There were no significant differences in the levels of FSH,LH,E2 and the antral follicle count of the patients between two groups 6 months and 1 year after operation(P>0.05).The incidence of perimenopausal symptoms of the patients in two groups 6 months and 1 year after operation showed no significant difference(P>0.05).The incidence of ovarian malignant tumor and ovarian benign tumor of the patients in two groups 6 months and 1 year after operation had no significantly differences (P>0.05). The incidence of pelvic inflammatory disease of the patients in control group was higher than that in prevention group(P<0.05). There were 2 patients diagnosed as tubal carcinoma in control group 1 year after operation, and the pathological findings showed the atypical cells in 2 patients in prevention group. Conclusion: Prophylactic bilateral salpingectomy during hysterectomy does not damage the ovarian function and can reduce the incidence of pelvic malignant/benign tumor and pelvic inflammatory disease.

Objective: To discuss the clinical characteristics,diagnosis and treatment experience of a case of lateral ectopic thyroid. Methods: The patient with lateral ectopic thyroid was diagnosed by thyroid color ultrasound and single-photon emission computed tomography(SPECT)-CT examination. The right thyroidectomy and the thyroid tissue in cervical section Ⅳ of the patient were removed by received the operation.The pathological examination after operation was performed.The daily usage of euthyrox after operation was 50 μg. Results: The patient recovered well,who accepted 10 months follow-up with normal life and stable condition,and had no recurrence and metastasis. Conclusion: The mechanism of lateral ectopic thyroidy is not clear yet,the surgical treatment can result in good effectiveness and better prognosis.

Objective: To analyze the clinical materials of 2 patients with lung cancer complicatedwithsyndrome of inappropriate secretion of antidiuretic hormone (SIADH),and to improve the clinical awareness and management. Methods: One patient was treated with lung cancer chemotherapy alone for 7 courses. The other patient received the strictly limited for liquid intake and tolvaptan by oral against hyponatremia without chemotherapy. Results: With the relapsing and advancing of lung cancer, the hyponatremia and its relevant symptoms of the first patient were poorly relieved. While the level of serum sodium of the second patient returned to the normal level 3 d after administration of tolvaptan. Conclusion: About one-third of the patients with hyponatremia have SIADH. The etiological treatment and symptomatic therapy of hyponatremia should be given to the patients in clinical treatment.

Objective: To discuss the screening results and clinical characteristics of children of Miao and Dong nationalities with mediterranean anemia in ethnic minority areas of Qiandongnan State of Guizhou Province, and to clarify the differences of the mediterranean anemia among different minorities. Methods: A total of 1 623 children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State were selected by multistage stratified random sampling method; quantitative analysis of HbA2 and HbF was used to screen the selected children with mediterranean anemia initially; phenol chloroform extraction method was applied to extract the DNA from the children with mediterranean anemia; ASO/RDB-PCR reverse dot blot hybridization method was used to analyze the gene characteristics of the children with mediterranean anemia. Results: A total of 1 623 children of Miao and Dong nationalities were selected as the subjects. Among 938 children with Miao nationality, there were 18 children with positive α-mediterranean anemia and 36 children with positive β-mediterranean anemia, and the positive detection rate was 1.92%. Among 685 children with Tong nationality, there were 13 children with positive α-mediterranean anemia and 24 children with positive β-mediterranean anemia, and the positive detection rate was 3.50%. The detection rates of composite of α-and β-mediterranean anemia in the children of Miao nationality and Tong nationality were 1.49% and 4.61%.There was no significant difference in the detection rates of different kinds of mealiterranean anemia between two nationalities (P<0.05). The major gene mutations in α-mediterranean anemia were——SEA/-αα and -α3.7, and the major gene mutations in β-mediterranean anemia were CD17/N and CD14-15/N, while the major gene types of the composite of α-and β-mediterranean anemia were——SEA/β41-42 and——SEA/β17. There was no difference in the positive rates of major gene types of different kinds of mediter ranean anemia between two nationalities(P<0.05). Conclusion: There is no difference in the positive rate of children of Miao and Dong nationalities with mediterranean anemia in minority areas of Qiandongnan State. CD17/N,——SEA/-αα and——SEA/β41-42 are the major gene types of α-, β-, and αβ-mediterranean anemia, respectively.