Objective:To establish the pancreatic carcinoma models of rats with dimethybenzanthracene(DMBA) embedding method, to discuss the treatment effect of Da Huang Mu Dan Tang (DHMDT) in the rats with pancreatic carcinoma and the protective effects on the liver and kidney function, and to clarify the mechanisms. Methods:Sixty-five SD male rats were divided into blank control (n=10) and modeling group (n=55). DMBA was embedded in the capsule of pancreas to establish the pancreatic carcinoma models of rats. After 3 months, 50 successfully modeling rats were randomly divided into model, 5-FU (intravennous injection, 12 mg·kg^-1), low, middle and high doses of DHMDT (intragastric administration, 20,40 and 80 mg·kg^-1) groups. After 10 weeks of administration, the levels of alanine aminotransferase(ALT), aspartate transaminase(AST), urea nitrogen(BUN), creatinine(Cr) and total bilirubin(TBiL) in serum of the rats in various groups were measured; the weights and the inhibitory rates of the pancreatic tumor were detected; TUNEL method was used to analyze the apoptotic rates of tumor tissue; the expression levels of SST2R, Bax and Bcl-2 genes in pancreatic tumor tissue of the rats were detected by Q-PCR method. Results:Compared with model group, after treated for 10 weeks, the levels of ALT, AST, BUN, Cr and TBiL in serum of the rats in low, middle and high doses of DHMDT groups were significantly decreased (P<0.05 or P<0.01); the weights of tumor of the rats in middle and high doses of DHMDT groups were decreased (P<0.05 or P<0.01); the apoptotic rates of the cells in pancreatic tumor tissue of the rats in low, middle and high doses of DHMDT groups were increased (P<0.05 or P<0.01); the expression levels of SST2R gene in pancreatic tumor tissue of the rats in low, middle and high doses of DHMDT groups were significantly increased (P<0.05 or P<0.01); the expression levels of Bax gene in pancreatic tumor tissue of the rats in middle and high doses of DHMDT groups were significantly increased(P<0.05 or P<0.01), while the expression levels of Bcl-2 in pancreatic tumor tissue of the rats were significantly decreased (P<0.05). Conclusion:DHMDT has the treatment effect in the rats with pancreatic carcinoma and can promote the apoptosis of tumor cells;DHMDT can protect the liver and kidney function of the rats with pancreatic carcinoma.

Objective:To explore the improvement effect of rhein lysinate (RHL) on the insulin resistance in the KK/HlJ diabetes mellitus (DM) mice induced by streptozotocin (STZ), and to elucidate its mechanism. Methods:The KK/HlJ diabetic mouse models were made by intraperitoneal injection of STZ (50 mg·kg^-1) and fed with DM diet.A total of 48 mice were divided into normal control group, model group, low dose of RHL group (25 mg·kg^-1) and high dose of RHL group (50 mg·kg^-1)(n=12). All the mice were treated for 16 weeks. The levels of fasting glucose (FBG),total cholesterol(TC) and triglyceride(TG) of the mice were measured by glucose oxidase method. HE staining was used to observe the morphology of pancreas tissue of the mice. The insulin level in pancreas islet tissue was detected by immunohistochemistry method. The levels of insulin and of C reactive protein(CRP) in serum and tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in liver tissue of the mice were detected by enzyme linked immunosorbent assay (ELISA) method. The expression levels of glycogen synthesis related genes (PI3K, AKT and GSK-3β) phosphorylation were detected by Western blotting method. Results:Compared with model group, the levels of FBG, TG and TC of the mice in low and high doses of RHL groups were significantly decreased (P<0.05), but there was no significant difference in the insulin level (P>0.05). Compared with model group, the levels of CRP in serum and the levels of TNF-α and IL-6 in liver tissue of the mice in low and high doses of RHL groups were significantly decreased (P<0.01). The HE staining results showed that compared with model group, the islet morphology of the mice in low and high doses of RHL groups was partially restored, and the occasional inflammatory infiltration was observed.The immunohistochemical staining results showed that compared with model group, the brown granular substance in the islets of the mice in low dose of RHL group was significantly reduced, which was disappeared in high dose of RHL group. Compared with model group, the expression levels of glycogen synthesis related genes (PI3K, AKT and GST-3β)phosphorylation of the mice in low and high doses of RHL groups were increased (P<0.01). Conclusion:RHL has improvement effect on the insulin resistance in the KK/HlJ DM mice induced by STZ,and the mechanism may be related to promoting the glycogen synthesis.

Objective:To explore the effect of 14-3-3ε protein on the localization of Cdc25B protein during the meiotic resumption of mouse oocytes, and to pay foundation for the further study on the molecular mechanism of 14-3-3εprotein in regulating the development of mouse oocytes. Methods:The Kunming genealology female mice aged 3 weeks were used to obtain the germinal vesicle (GV)-stage oocytes after superovulation. The GV-stage oocytes were divided into non-injection group,control siRNA injection group and 14-3-3ε siRNA injection group.The pmax-FP-Red-HA-14-3-3ε expression vector was constructed. Indirect immunofluorescence was used to observe the colocalization of 14-3-3ε protein and Cdc25B protein in the mouse oocytes; direct immunofluorescence was used to observe the subcellular localization of 14-3-3ε protein and Cdc25B protein in the mouse oocytes;14-3-3ε siRNA was microinjected into the GV-stage oocytes;the morphology was observed under phase-contrast microscope; the germinal vesicle breakdown(GVDB) rates of the mouse oocytes were calculated;the expression level of 14-3-3ε protein and the relative expression level of Cdc2-pTyr15 protein were observed by Western blotting method;the matuation-promoting factor (MPF) activity in the oocytes was measured by autoradiography. Results:The indirect immunofluorescence and direct immunofluorescence results showed that the 14-3-3ε protein and wild Cdc25B protein were co-localized in the cytoplasm; Cdc25B was translocated from the cytoplasm to the nucleus shortly before GVBD. When the Ser321 of Cdc25B protein turned into Ala, the expression level of 14-3-3ε protein was decreased. None of the oocytes in non-injection group and control siRNA injection group were able to undergo GVBD until at least 24 h after injection,there was no significant differences in the rate of GVBD between non-injection group and control siRNA injection group (P>0.05);the GVBD rates of oocytes in 14-3-3ε siRNA injection group at 22 and 24 h after injection were significantly higher than those in non-injection group and control siRNA injection group(P<0.01);the rate of oocytes progressed to metaphaseⅡ(MⅡ) in 14-3-3ε siRNA injection group at 24 h after injection was significantly higher than those in non-injection group and control siRNA injection group(P<0.01). Conclusion:Ser321 might be involved in the process of regulating the subcellular localization of Cdc25B by 14-3-3ε protein in the meiotic resumption of mouse oocytes.

Objective:To investigate the inhibitory effect of T-2 toxin on proliferation of the human hepatocellular carcinoma HepG2 cells and its promotion effect of apoptosis. Methods:The HepG2 cells in the logarithmic phase were selected and divided into control group (without T-2 toxin) and experimental groups (given 0.25, 2.50, 25.00, 250.00 and 2 500.00 μg·L^-1T-2 toxin). After 24 h treatment, the morphology of cells was observed under inverted microscope; the inhibitory rate of proliferation of cells was determined by MTT assay; the cell cycle and apoptotic rate of cells were analyzed by flow cytometry; Hochest 33258 staining was used to observe the apoptotic morphology of cells; the activity of caspase-3 in HepG2 cells was detected. Results:After treated for 24 h, the inverted microscope observation results showed that the number of the cells in experimental groups was decreased significantly and the cells shrank and deformed. The MTT results showed that compared with control group, the inhibitory rates of proliferation of the cells in experimental groups were increased (P<0.01). The flow cytometry results showed that compared with control group, the percentage of the cells in SubG₁ phase in experimental group was significantly increased, and the apoptotic rates of the cells in experimental groups were significantly increased. The Hoechest 33258 staining results showed that the chromatin condensation was observed in the cells in experimental groups, and the nuclei were dense and stained. Compared with control group, the activities of intracellular caspase-3 of the cells in experimental groups were significantly increased (P<0.01). Conclusion:T-2 can inhibit the proliferation of human hepatocellular carcinoma HepG2 cells, and induce the apoptosis.

Objective:To investigate the effect of cartilage tissue engineering scaffold PVA/ι-CA on the biological behavior of the ATDC-5 cells,and to evaluate its feasibility on constructing tissue engineering cartilage. Methods:The polyvinyl alcohol(PVA) and carrageenan were used to make the composite scaffold material PVA/ι-CA according to a certain proportion by physical blending technology and repeated freezing thawing method,and the porosity and pore size of PVA/ι-CA were detected.The ATDC-5 cells were seeded into the composite scaffold and its growth was observed; the expressions of collagen type Ⅱ in the ATDC-5 cells were tested by immunohistochemical staining and immunofluorescence staining;the morphology of the ATDC-5 cells was confirmed by Toluidine blue staining. The growth and secretion of extracellular matrix of the ATDC-5 cells were observed under scanning electron microscope(SEM);the proliferative rates of ATDC-5 cells in composite scaffold materials in negative control group (added with DMEM culture media) and experimental group(added with DMEM contain scaffold) were determined by MTT assay. The composite scaffolds were implanted subcutaneously in the SD rats.The histocompatibility and vascularization in vivo of the composite scaffolds were evaluated. Results:The average porosity of cartilage tissue engineering scaffold PVA/ι-CA was (86.88±3.88)%,and the average pore size was 20-40 μm.The HE staining results showed that the ATDC-5 cells grew well with the polygon and plumpness morphology. All the samples were stained positive for collagen type Ⅱ by immunohistochemistry and immunofluorescence staining, which verified the normal phenotype of chondrocytes on the scaffolds. All the sample were stained positive for toluidine blue staining, which verified ECM deposition of the ATDC-5 cells on the scaffolds.The number of the positive cells was significantly increased with the prolongation of time. After cultured for 7 d,few of the ATDC-5 cells presented polygonal; after cultured for 14 d,the ATDC-5 cells distributed more densely,and contacted with each other on the scaffold; after cultured for 21-28 d,the ATDC-5 cells filled the interconnected pores of the scaffolds, synthesizing a significant amount of neo-formed ECM. The proliferation of ATDC-5 cells in PVA/ι-CA grew fast during 7-14 d,and it became slow during 21-28 d;the difference was not statistically significant compared with control group(P>0.05).The subcutaneous implantation results showed the inflammatory reactions were slight at the early stage and eviated gradually,there was an increasing angiogenesis at the late stage,and the degradation and absorption of the meterial were slight. Conclusion:PVA/ι-CA composite material will be an ideal material for the cartilage tissue engineering.

Objective:To study the influence of proteasome inhibitor lactacystin(LAC)and carboplatin in proliferation and apoptosis of the human ovarian cancer SKOV3 cells in vitro, and to clarify the mechanisms. Methods:The SKOV3 ovarian cancer cells were cultured in vitro; 0, 2.5, 5.0, 10.0 and 20.0 μmol·L^-1 LAC were used to intervent the SKOV3 cells for 48 h;5 μmol·L^-1 LAC was used to intervent the SKOV3 cells for 0, 24, 48, and 72 h; the SKOV3 cells were divided into control group (treated without medical intervention), LAC group (treated with 5 μmol·L^-1 LAC), carboplatin group(treated with 10, 20, 40 and 80 μmol·L^-1 carboplatin), LAC and carboplatin group(treated with 5 μmol·L^-1 LAC and 10, 20, 40, and 80 μmol·L^-1 carboplatin, respectively).MTT method and FCM were used to detect the inhibitory rates of proliferation and apoptotic rates of the SKOV3 cells in various groups. Results:The MTT test results showed that the proliferation of the SKOV3 cells were inhibited with the prolongation of time and increasing of LAC concentration;the half inhibitory concentration(IC₅₀) of LAC at 48 h was 5.36 μmol·L^-1; compared with carboplatin group, the inhibitory rates of proliferation of SKOV3 cells in LAC and carboplatin groups were significantly increased(P<0.05). The IC₅₀ of carboplatin was dropped from 58.08 μmol·L^-1 to 18.37 μmol·L^-1.The FCM results showed that with the prolongation of treated time of LAC, the apoptotic rates of SKOV3 cells were increased;compared with carboplatin group and LAC group, the apoptotic rate of cells in LAC and carboplatin group was increased(P<0.05). Conclusion:LAC can inhibit the proliferation of the ovarian cancer SKOV3 cells and induce the apoptosis, and LAC can enhance the inhibitory effect of proliferation of carboplatin on the ovarian cancer SKOV3 cells.

Objective:To investigate the lipid-lowering effect and antioxidant activity of polysaccharide from Schisandra Chinensis (SCP) in the rats with non-alcoholic fatty liver disease (NAFLD) induced by high-fat diet, and to provide a theoretical basis for the development and utilization of Schisandra Chinensis. Methods:A total of 32 male Wistar rats were selected. Sixteen from the 32 rats were randomly selected and divided into normal control group (intragastrical administration of water, combined with normal diet, n=8) and SCP group (intragastrical administration of 50 mg·kg^-1SCP, combined with normal diet, n=8). The remaining 16 rats were fed with high-fat diet for 4 weeks and the confirmed NAFLD rat models were set up. A total of 16 NAFLD rats were randomly divided into NAFLD group (intragastrical administration of water, combined with high-fat diet, n=8) and NAFLD+SCP group (intragastrical administration of 50 mg·kg^-1SCP, combined with high-fat diet, n=8). After treated for 12 weeks, the body weights of all the rats were weighed and the liver index was calculated. The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum of all the rats were determined. The levels of TC and TG in liver tissue of the rats were measured by enzymatic method.The malondialdehyde (MDA) levels and superoxide dismutase (SOD) activities in serum and liver tissue and the glutathione(GSH) levels in liver tissue of the rats were analyzed by TBA, xanthinoxidase and microscale enzyme methods, respectively. HE staining was used to observe the pathomorphology of liver tissue of the rats. Results:Compared with normal control group, the liver index of the rats in NAFLD group was increased (P < 0.01); the levels of TC, TG, LDL-C, ALT and AST in serum of the rats were increased(P < 0.01), the levels of TC and TG in liver tissue of the rats were increased (P < 0.01), the MDA level was increased(P < 0.01) and the SOD activity was decreased(P < 0.01), and the GSH levels in liver tissue and serum were decreased(P < 0.01). Compared with NAFLD group, the body weight and liver index, serum levels of TC, TG, LDL-C, ALT and AST of the rats in NAFLD+SCP group were decreased (P < 0.05), the levels of TC and TG in liver tissue were decreased(P < 0.01), the MDA level was decreased (P < 0.01),the SOD activities in serum and liver tissue were increased(P < 0.01), and the level of GSH in liver tissue was increased(P < 0.01). The HE staining results showed that the structure of hepatic lobules of the rats in NAFLD group was disordered and showed significant hepatic steatosis,and the hepatic steatosis in hepatic lobules of the rats in NAFLD+SCP group was significantly reduced. Conclusion:SCP has a regulation effect in the NAFLD rats induced by high-fat diet, and its mechanism may be related to the anti-oxidative stress.

Objective:To discuss the prokaryotic expression system and purification conditions of Toxoplasma gondii profilin-like protein (TgPRF), and to provide basis for the study on anti-tumor immuno-adjuvant. Methods:The coding region of TgPRF gene was amplified with a pair of specific primers which were designed according to the cDNA oftachyzoites of Toxoplasma gondii RH strain. The PCR products were cloned into the pET-28a(+) vector after double enzyme digestion. The recombinant pET28a(+)-TgPRF plasmid was transformed into E.coli DH5α cells. The positive clones were selected by the double restrictive enzyme digestion and sequencing. The correct pET28a (+) -TgPRF plasmid was transformed into E.coli BL21(DE3) and induced for 4 h by IPTG. The expression of recombinant TgPRF protein was analyzed by SDS-PAGE method; the expression of recombinant protein His-profilin was detected by Western blotting method. Results:The length of product of PCR was 492 bp. The recombinant plasmid pET28a-TgPRF was confirmed by double restriction enzyme digestion and sequencing. The SDS-PAGE results showed that the target protein was expressed in E.coli BL21 (DE3) in bacteria supernatant. The purified TgPRF protein was obtained by Ni-NTA agarose gel column chromatography with the purity>90%. The Western blotting results revealed that the recombinant TgPRF protein could be recognized by Anti-His antibody. Conclusion:The recombinant plasmid pET28a-TgPRF is successfully constructed, and the TgPRF protein is obtained with the soluble prokaryotic expression.

Objective:To observe the improvement of acidic polysaccharose of Schisandrae Chinensis (SCP-A) on the learning and memory functions of Alzheimer's disease (AD) model mice induced by Aβ_25-35, and to clarify the related mechanisms. Methods:A total of 75 male C57BL mice were randomly divided into control group (injected with saline in intracerebroventricular, given distilled water intragastrically), model group (injected with Aβ_25-35 intracerebroventricularly, given distilled water intragastrically), and 5, 10,20 mg·kg^-1 SCP-A groups (injected with Aβ_25-35 intracerebroventricularly, given SCP-A intragastrically)(n=15). The agents were administered once daily for 14 d, in which Aβ_25-35 was injected intracerebroventricularly on the 8th day. On the 15th day after administration, step-through test and Morris water maze test were used to detect the escape latency, number of errors,and time of finding platform, number of passing platforms,dwell time in target quadrant of the mice; Western blotting method was used to detected the levels of Tau,GSK-3β and their phosphorylated proteins in hippocampus tissue of the mice. Results:The step-through test results showed that compared with control group,the escape latency of the mice in model group was obviously shortened and the number of errors was significantly increased(P<0.01); compared with model group, the escape latencies of the mice in 10 and 20 mg·kg^-1 SCP-A groups were significantly prolonged (P<0.01), the number of errors was significantly decreased (P<0.05 or P<0.01). The Morris water maze test results showed that compared with control group, the time of finding platform of the mice in model group was significantly prolonged,the number of passing platform and dwell time in target quadrant were significantly reduced(P<0.01); compared with model group, the time of finding platform of the mice in 10 and 20 mg·kg^-1 SCP-A groups was significantly shortened(P<0.01),and the number of passing platform and the dwell time in target quadrant were significantly increased (P<0.01). The Western blotting results showed that compared with model group, the expression levels of phosphorylated protein Tau Ser199, Tau Ser396 and Tau Ser404 in hippocampus of the mice in 20 mg·kg^-1 SCP-A group were significantly decreased(P<0.05), and the expression level of GSK-3β was significantly decreased(P<0.01); the expression level of phosphorylated protein GSK-3β Tyr216 was significantly decreased, and the expression level of phosphorylated protein GSK-3β Ser9 was significantly increased (P<0.05). Conclusion:SCP-A has an anti-AD effect, which is related to regulating the activity of GSK-3β to reduce the level of phosphorylated protein Tau in the hippocampus tissue of the mice.

Objective:To detect the expressions of ovarian function-related proteins in the ovarian follicles in different development stages, and to explore their roles and significances in the regulation of follicular development and ovarian function. Methods:The ovarian tissue containing primordial, primary, secondary follicles, mature follicles,atretic follicles and corpus albicans of the woman in the childbearing age was selected.Immunohisto chemical analysis was used to detect the expressions of Glyoxal enzyme I (glyoxalase I), ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), heat shock protein 27 (HSP27), serum amyloid P (SAP) proteins in the ovarian follicles in different stages.The expression intensities of these proteins were compared. Results:The expression intensities of Glyoxalase I and UCH-L1 in the granulosa cells and theca cells of secondary follicles and marure follicles were strong, which were higher than those in the cytoplasm of primordial and primary follicles. The expression intensities of HSP27 in the cytoplasm of primordial and primary follicles were strong, which were higher than those in the granulosa cells and theca cells of secondary follicles and marure follicles.The expression intensities of Glyoxalase I and HSP27 in the atretic follicles were strong,which were higher than that in the growth follicles.The expressions of SAP were positive in the primordial, primary follicles, secondary and mature follicles and the expression intensities were not different. The expressions of four proteins in the corpus albicans did not express. Conclusion:The high expression of UCH-L1 and low expression of HSP27 are associated with the mature and development of follicles; the high expressions of Glyoxalase I and HSP27 are associated with the follicular atresia and ovarian failure.

Objective:To investigate the effect of pilose antler polypeptide combined with Schwann cells modified by glial cell line-derived neurotrophic factor(GDNF) gene on the proliferation of human bone marrow mesenchymal stem cells(BMSCs) in vitro. Methods:According to the conventional method,the bone marrow (10 mL) was extracted from the healthy volunteers and was inoculated into the culture flask. The primary cultured cells were completely fused. The BMSCs were harvested at the 3rd generation and the cells were adjusted to 5×10⁶ mL^-1. 4 μL GDNF gene modified Schwann cells was added into GDNF group, 4 μL (10 mg·L^-1) PAP combined with GDNF gene modified Schwann cells was added into combination group, and only same amount of medium was added into control group. The proliferative activities, cell nuclear antigen (PCNA) levels and apoptotic rates of BMSCs in various groups were detected by enzyme-linked immunosorbent assay, ELISA method and Annexin Ⅴ-FIFC/PI cell apoptosis detection kit,respectively. Results:After primary culture for 48 h, most of the cells adhered to the wall, and the morphology of the cells changed into polygonal shape and few of them showed spindle. The passaged cells showed spindle spindle,the cells were confirmed as BMSCs, and all of them were non-hematopoietic stem cells. Compared with control group, the proliferative activities and the PCNA level of the BMSCs in GDNF group and combination group were increased(P<0.05) and the apoptotic rates were decreased(P<0.05); compared with GDNF group, the proliferative activity and the PCNA level of the BMSCs in combination group were increased (P<0.05) and the apoptotic rate was decreased (P<0.05). Conclusion:PAP combined with Schwann cells modified by GDNF gene can promote the proliferation of human BMSCs in vitro.

Objective:To study the effects of Ginkgo Biloba extract(GBE) on the differentiation and bone resorptionof the osteoclasts, and to clarify their mechanisms. Methods:The RAW264.7 cells were cultured in vitro, then were treated with receptor activator for nuclear factor-κB ligand (RANKL) and different concentrations of GBE. The cells were divided into blank control group(0 μg·L^-1 RANKL), RANKL group(100 μg·L^-1 RANKL), RANKL+75 mg·L^-1 GBE group,and RANKL+150 mg·L^-1 GBE group.The morphology and number of osteoclasts were assessed with TRAP staining assay, and bone resorption of GBE was examined with bone resorption pits assay. Flow cytometry was applied to analyze the the apoptotic rate of RAW264.7 cells and the cell cycle;the expression levels of nuclear factor of activated T cells 1 (NFATc1), DC-STAMP, Casthepin K,matrix metalloprotein 9(MMP-9), Bcl-2, Bax,P27 and Cyclin-D1 in the RAW264.7 cells were analyzed by RT-PCR method. Results:Compared with blank control group, the number of osteoclasts in RANKL group were significantly increased(P<0.01); compared with RANKL group,the number of osteoclasts in RANKL+75 mg·L^-1 GBE and RANKL+150 mg·L^-1 GBE groups were significantly decreased (P<0.05). Compared with blank control group,the area of bone resorption pit of bone slice in RANKL group was significantly increased(P<0.05);compared with RANKL group, the areas of bone resorption pit of bone slice in RANKL+75 mg·L^-1 GBE and RANK+150 mg·L^-1 GBE groups were significantly decreased (P<0.05). Compared with blank control group, the apoptotic rates of the RAW264.7 cells in 75 and 150 mg·L^-1 GBE groups were increased, and the expression levels of Bcl-2 in the RAW264.7 cells were significantly decreased and the expression levels of Bax were significantly increased (P<0.05). Compared with RANKL group, the G₀-G₁ phase arrest of the RAW264.7 cells in RANKL+75 mg·L^-1 GBE and RANKL 150 mg·L^-1 GBE groups were shortened;the expression levels of P27 in the RAW264.7 cells were significantly decreased and the expression levels of Cyclin-D1 were significantly increased (P<0.05). Compared with blank control group, the expression levels of NFATc1, DC-STAMP, Casthepin K and MMP-9 in the RAW264.7 cells in RANKL group were significantly increased(P<0.05); compared with RANKL group, the expression levels of NFATc1, DC-STAMP, Casthepin K and MMP-9 in the RAW264.7 cells in RANKL+75 mg·L^-1 GBE and RANKL+150 mg·L^-1 GBE groups were significantly decreased (P<0.05). Conclusion:GBE could inhibit the differentiation and bone resorption of the osteoclasts, and their mechanisms may be related to promoting the apoptosis of RAW264.7 and shortening the G₀-G₁ phase of the RAW264.7 cells.

Objective:To investigate the effect of alteplase on the expressions of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells after thrombolysis in the rats with acute cerebral infarction and its mechanism, and to provide experimental evidence for its clinical application. Methods:The models of acute thrombosis of middle cerebral artery of the rats were established.Fifty-four SD rats were randomly divided into sham operation group, model group and alteplase group(n=18).The ultrastructure of brain endothelial cells of the rats was observed under transmission electron microscope.The expression levels of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells of the rats were detected by immunoflurorescence and Western blotting methods. Results:The transmission electron microscope results showed that the brain volume in the ischemic area of the rats in model group was significantly increased and the endothelial cells were swollen, some of the cortical and surrounding brain tissues had obvious boundaries,and the thickness of the basement membrane was uniform and the tightly connected structure was very loose and disappeared; the swelling condition of the capillary endothelial cells in infarcted area of the rats in alteplase group was significantly reduced and the thickness of the basement membrane was improved, and most of the tightly connected structures between the brain capillary endothelial cells and the endothelial cells were loose and the fracture was lost and the structure disappeared. The immunofluorescence results showed that the expressions of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells of the rats in alteplase group were significantly improved compared with model group. The Western blotting results showed the expression levels of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells of the rats in model group were significantly decreased compared with sham operation group(P<0.01);compared with model group,the expression levels of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells of the rats in alteplase group at different time points were increased(P<0.01). Conclusion:Alteplase can improve the structure of brain endothelial cells in the rats with acute cerebral infarction, and the mechanism may be related to decreasing the expressions of Claudin-1 and Claudin-5 proteins in the vascular endothelial cells of the rats induced by alteplase.

Objective:To investigate the effects of cerebral ischemic postconditioning (CIP) on the expressions of Cyclin D1 and CDK4 proteins in hippocampal neurons of the rats with global cerebral ischemia and reperfusion (I/R) based on Notch1 signaling pathway,and to explore the mechanisms. Methods:A total of 128 healthy male SD rats were randomly divided into sham operation group, I/R group (modified Pulsinelli four vessel occlusion method), CIP group(repeated 3 times of reperfusion or blocking blood vessel before complete reperfusion) and DAPT group(intraperitoneal injection of 10 mg·kg^-1·d^-1 DAPT 3 h before CIP),and there were 32 rats in each group. HE staining was used to observe the morphology of the neurons at 6, 24, 48 and 72 h after ischemia;immunohistochemical staining was used to observe the expressions of Cyclin D1, CDK4 and Notch1 in the hippocampus of the rats;Western blotting method was used to observe the expression levels of Cyclin D1, CDK4 and Notch1 proteins in the hippocampus of the rats at different time points. Results:The HE staining results showed that compared with sham operation group, the structure of neurons in hippocampus area of the rats in I/R group was damaged and the survival rate of neurons was significantly decreased(P<0.05).Compared with I/R group, the survival rates of neurons in the hippocampus area of the rats in CIP group were significantly increased at the corresponding time(P<0.05).Compared with CIP group, the survival rates of neurons in hippocampus area of the rats in DAPT group at the corresponding time were significantly decreased (P<0.05).The immunohistochemical staining results showed that compared with sham operation group,the number of cells with positive Notch1, Cyclin D1 and CDK4 in I/R group was increased (P<0.05).Compared with I/R group, the number of Cyclin D1 and CDK4 positive cells in CIP group was decreased (P<0.05), and the number of Notch1 positive cells of was significantly increased (P<0.05).Compared with CIP group, the number of Cyclin D1 and CDK4 positive cells in DAPT group was increased (P<0.05), and the number of Notch1 positive cells was significantly decreased(P<0.05).The Western blotting results showed that compared with sham operation group, the expression levels of Notch1, Cyclin D1 and CDK4 proteins in the hippocampus of the rats in I/R group were increased (P<0.05);compared with I/R group, the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in CIP group were significantly decreased (P<0.05), and the expression level of Notch1 protein in the hippocanpus of the rats was significantly increased (P<0.05);compared with CIP group, the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in DAPT group were increased (P<0.05), and the expression level of Notch1 protein was significantly decreased (P<0.05). Conclusion:CIP may play an important role in protecting the neurons by increasing the activity of Notch1 and inhibiting the expressions of Cyclin D1 and CDK4.

Objective:To investigate the promotion effect of 4,5,6,7-tetrabromobenzotriazole (TBB) on the apoptosis of human colon cancer SW480 cells, and to explore its possible mechanism. Methods:The human colon cancer SW480 cells at logarithmic growth phase were divided into control group (0 μmol·L^-1 TBB) and experiment group (1, 3, 10, 30, and 100 μmol·L^-1 TBB). The viability of cells was measured by MTT assay; the apoptotic rate of the SW480 cells and the level of intracellular reactive oxygen species (ROS) were analyzed by Annexin Ⅴ-FITC/PI double staining and flow cytometry. The expression levels of anti-apoptotic proteins p-Akt and Bcl-2, and pro-apoptotic proteins Bad, pro-caspase-9 and cleaved-caspase-3 were detected by Western blotting method. Results:The MTT results showed that the viabilities of SW480 cells in experiment group were decreased in a dose-dependent manner, which were lower than that in control group (P<0.05).The Annexin Ⅴ-FITC/PI double staining and flow cytometry results showed that the apoptotic rates of SW480 cells in experiment group (3, 6, 12, and 24 h)were significantly higher than that in control group (0 h) (P<0.05). The flow cytometry results showed the levels of ROS in SW480 cells after treated with TBB for 3, 6, 12 and 24 h were higher than that in 0 h group (P<0.05). The Western blotting results showed that the expression levels of anti-apoptotic proteins p-Akt and Bcl-2 in SW480 cells in experiment group (3, 6, 12, and 24 h) were decreased obviously, whereas the expression levels of the pro-apoptotic proteins Bad and cleaved-caspase-3 were increased and the expression level of pro-caspase-9 was decreased compared with those in control group (0 h) (P<0.05). Conclusion:TBB could inhibit the cell proliferation and induce the apoptosis of human colon cancer SW480 cells,and its mechanism may be related to the inhibition of the activity of Akt and the promotion of the level of intracellular ROS.

Objective:To investigate the improvement effects of duodenal-jejunal bypass (DJB) on the blood glucose homeostasis, insulin resistance and inflammation of the obese type 2 diabetic(T2DM) ZDF rats, and to discuss its possible mechanism. Methods:A total of 20 ZDF rats were randomly divided into DJB operation group and sham operation group (n=10). There were 8 rats survived in each group after operation. The level of blood glucose (FBG) was detected by Roche glucose meter at 1 week before operation, 2 weeks, 4 weeks and 6 weeks after operation; the fasting serum insulin level of the rats was measured by ELISA kit; the insulin sensitivity index (HOMA-ISI) and insulin resistance index (HOMA-IR) were calculated. The rats were executed 6 weeks after operation. HE staining was used to observe the morphology of the inflammatory cells in BP limb of the rats; the expression levels of AMPK and pAMPK in BP lamb of the rats were observed by immunohistochemical staining;the expression levels of interleukin 1β (IL-1β), interleukin 6(IL-6), tumor necrosis factor α (TNF-α), nuclear factor κB (NF-κB), and interleukin 10 (IL-10) mRNA of the rats were detected by QRT-PCR method. Results:From the 2nd week after operation,compared with before operation,the FBG levels of the rats in DJB operation group were decreased(t=3.798, P<0.05);compared with sham operation group,the FBG level of the rats in DJB operation group was decreased (t=3.205, P<0.05). Six weeks after operation, compared with sham operation group, the HOMA-IR of the rats in DJB operation group was significantly decreased (t=4.441, P<0.05) and the HOMA-ISI was significantly increased (t=-8.65, P<0.05).The HE staining results showed that compared with sham operation group, the morphology of the inflammatory cells in BP limb of the rats in DJB operation group was significantly improved.The QRT-PCR results showed that the expression levels of IL-1β, IL-6, TNF-α and NF-κB of the rats in DJB operation group was significantly decreased compared with sham operation group (P<0.05), while the expression level of IL-10 was significantly increased(P<0.05). The immunohistochemical test results showed that the expression levels of AMPK and pAMPK in BP lamb of the rats in DJB operation group were increased compared with sham operation group. Conclusion:DJB can significantly improve the blood glucose homeostasis and insulin resistance in the T2DM rats, and its mechanism may be related to the decreased expressions of inflammatory factors and the activation of AMPK molecules in BP lamb of the T2DM rats.

Objective:To explore the induction of oxaziridine on the apoptosis of human cervical cancer HeLa cells and its mechanism. Methods:The human cervical cancer HeLa cells were treated with 0,0.5,1.0,5.0 and 10.0 μmol·L^-1 oxaziridine for 24 h and used as control group and different concentrations of experiment groups. The morphology of HeLa cells was observed under inverted microscope; the surival rates and inhibitroy rates of proliferation of the HeLa cells in various groups were measured by MTT assay. The HeLa cells were treated with 0,0.5,1.0 and 5.0 μmol·L^-1 oxaziridine for 12 h, and Hoechst 33258 staining was used to detect the apoptotic morphology of the cells; Western blotting method was used to detect the relative expression levels of Bcl-2 and Bax proteins. Results:The MTT assay results showed that the survival rates of the HeLa cells in 0.5,1.0,5.0 and 10.0 μmol·L^-1 groups were decreased and the inhibitory rates of proliferation of the HeLa cells were increased(P<0.05 or P<0.01)compared with control group, and the half inhibitory concentration(IC₅₀)was 2.6 μmol·L^-1. The cells in control group(0 μmol·L^-1 oxaziridine group) were in good condition, and the volumes of the HeLa cells in 0.5, 1.0,5.0 and 10 μmol·L^-1 groups were significantly reduced and the intercellular junction also disappeared. The Hoechst 33258 staining results showed the enhanced staining in the cells treated with oxaziridine.The Western blotting results showed that compared with control group(0 μmol·L^-1 oxaziridine group), the relative expression levels of Bcl-2 protein in the HeLa cells in 0.5, 1.0 and 5.0 μmol·L^-1 oxaziridine groups were significantly decreased(P<0.05 or P<0.01), and the relative expression levels of Bax protein were significantly increased (P<0.05 or P<0.01). Conclusion:Oxaziridine can effectively inhibit the proliferation of HeLa cells in vitro and induce the apoptosis of HeLa cells.

Objective:To investigate the influencing factors of sulfated modification of Bupleurum chinense polysaccharides (BCP), and to elucidate the possible mechanism of improving the antioxidant ability of sulfated BCP (S-BCP). Methods:BCP was sulfated by chlorosulfonic acid-pyridine method. The degree of substitution (DS) of S-BCP was observed by adjusting the volume ratios of chlorosulfonic acid to pyridine(1:2,1:4, and 1:8). The structures of BCP and S-BCP were analyzed by infrared (IR) spectroscopy, the morphology of BCP and S-BCP were observed under scanning electron microscopy (SEM). The antioxidant model was established by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging. The experiment was divided into positive control group, BCP group and S-BCP group, and the scavenging rates of DPPH free radical in various groups were compared. Results:When the volume ratio of chlorosulfonic acid to pyridine was 1:4, the reaction time was 2 h and the reaction temperature was 60℃, the maximum sulfur content percentage of S-BCP was 18.62% and the DS was the highest (DS=2.32). Compared with BCP group,the scavenging rate of DPPH free radical of S-BCP was significantly increased (P<0.05). Conclusion:The volume ratio of chlorosulfonic acid to pyridine can affect the DS of S-BCP. The sulfated modification can increase the anti-oxidant capacity of BCP by changing its physic-chemical characters and spatial conformation.

Objective:To explore the efficacy and adverse effects of docetaxel combined with capecitabine on the basis of anthracycline in the treatment of breast cancer patient on the basis of anthracycline, and to provide evidence-based medicine evidence for the clinical application of capecitabine. Methods:PubMed, EMBase, CNKI, VIP and Wanfang database were used to search the randomized controlled trials in which the breast cancer patients were treated by docetaxel combined with capecitabine and docetaxel alone. The qualities of the included studies were assessed by the modified Jadad scale. The effective data were extracted and the pathologic complete remission (pCR), recurrence-free survival (RFS), disease-free survival (DFS) and safety were evaluated by RevMan 5.3 software. Results:A total of 10 articles were included, including 12 012 patients. The Meta-analysis results showed that there were no statistically significant differences in pCR (RR=1.04, 95%CI:0.91-1.18, P=0.59),RFS(RR=0.86, 95%CI:0.71-1.03,P=0.10) and DFS(RR=0.98, 95%CI:0.86-1.11,P=0.71) between docetaxel combined with capecitabine and docetaxel alone. The clinical safety analysis results showed that the patients treated with docetaxel combined with capecitabine were susceptible to hand-foot syndrome (OR=7.13, 95%CI:4.76-10.69, P<0.001) and stomatitis (OR=1.93, 95%CI:1.10-3.40, P=0.02). Conclusion:On the basis of anthracycline treatment, the efficacies of docetaxel combined with capecitabine and docetaxel alone in the treatment of the breast cancer patients are similar. The former is more likely to cause the hand-foot syndrome and stomatitis.

Objective:To explore the expression of Bcl related anti-apoptotic protein3(BAG3 protein) in colon cancer and its clinical significance, and to clarify the relationship between its expression and the occurrence and development of colon cancer. Methods:A total of 90 cases of colon cancer tissue and paracancerous tissue were selected, the expressions of BAG3 protein in different tissues were detected by tissue microarray and immunohistochemical SP method; Kaplan-Meier method, univariate analysis and multivariate analysis were used to investigate the relationship between the clinicopathological parameters and the prognosis of the patients with colon cancer. Results:The expression of BAG3 protein in the colon cancer tissue was high,and the positive expression rates of BAG3 protein in the colon cancer tissue and paracancerous tissue were 37.8% and 0.01%, respectively,and the difference was significant(P < 0.05).The expression of BAG3 protein was associated with gender and tumor size(P < 0.05),but there was no correlation between the expression of BAG3 protein in the colon cancer tissue and age,pathologic grade,TNM stage,and lymph node metastasis (P > 0.05).The Kaplan-Meier results showed that the expression of BAG3 protein in the colon cancer tissue was not related with the prognosis of the patients and the survival rates of the younger patients and the higher TNM staging patients were high. Conclusion:The expression of BAG3 protein in the colon cancer tissue is high, and its expression is associated with the tumor size; BAG3 protein may act as a tumor marker and therapeutic target for colon cancer.

Objective:To investigate the feasibility of detection of serum pepsinogen (PG), gastrin-17 (G-17) and Helicobacter pylori antibody (anti-HP) in the gastric cancer screening, and to elucidate its clinical value. Methods:A total of 208 patients with early gastric cancer were selected as observation group; at the same time, 208 healthy examinees were regarded as blank control group. The levels of PGⅠ, PGⅡ and G-17 of the subjects in two groups were detected by enzyme-linked immunosorbent assay and latex-enhanced immunoturbidimetry; the positive rates of anti-HP were detected by ¹³C urea breath test; the serum levels of PG Ⅰ, PG Ⅱ and PG Ⅰ/PG Ⅱ ratio of the anti-HP positive subjects (anti-HP positive group) and anti-HP negative subjects (anti-HP negative group) were compared and analyzed. Results:Compared with blank control group, the serum PGⅠ/PGⅡ ratio and PGⅠ level of the subjects in observation group were significantly decreased(P<0.01), and the levels of PGⅡ,G-17and the positive rate of anti-HP were increased(P<0.01).Compared with anti-HP negative group, the serum PGⅠ/PGⅡ ratio and PGⅠ level of the subjects in anti-HP positive group were significantly decreased(P<0.01).The levels of PGⅡ and G-17 were significantly increased (P<0.01). Conclusion:The detection of positive rate of anti-HP combined with the ratio of PGⅠ/PGⅡ has important clinical significance in gastric cancer screening.The levels of G-17 and PG Ⅱ in the epithelial neoplasia lesion tissue can be used as indicators of gastric precancerous lesions.

Objective:To investigate the expression of transmembrane 4 super family 1(TM4SF1) in breast cancer tissue,and to elucidate its clinical significance and explore the related molecular biological mechanisms. Methods:A total of 190 cases of human breast cancer, 110 cases of paracancerous tissue and 110 cases of normal breast tissue were collected. Immunohistochemistry was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue, paracancerous tissue, and normal breast tissue; Western blotting method was used to detect the expression levels of TM4SF1 in breast cancer tissue, paracancerous tissue, and normal breast tissue; RT-PCR method was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue, paracancerous tissue, and normal breast tissue. The positive expression rates of TM4SF1 in breast cancer tissue of the breast cancer patients with different clinicopathological features were detected. Results:The positive expression rate of TM4SF1 in the breast cancer tissue was significantly higher than those in paracancerous tissue and normal breast tissue (P < 0.05); there was no significant difference in the positive expression rates of TM4SF1 between paracancerous tissue and normal breast tissue (P=0.531);the expression of TM4SF1 was not correlated with age, but was closely correlated with tumor size, differentiation degree, lymph node metastasis and tumor stage (P < 0.05);the positive expression rate of TM4SF1 in basal like breast cancer tissue was higher than those in the other three types of tissues (P < 0.05). The results of Western blotting showed that the expression level of TM4SF1 in breast cancer tissue was higher than those in paracancerous tissue and normal breast tissue (P < 0.05),but there was no significant difference in the expression level of TM4SF1 between paracancerous tissue and normal breast tissue(P > 0.05). The results of RT-PCR showed that the expression level of TM4SF1 mRNA in breast cancer tissue was higher than those in the paracancerous tissue and normal breast tissue(P < 0.01);there was no significant difference in the expression level of TM4SF1 mRNA between paracancerous tissue and normal breast tissue(P > 0.05). Conclusion:The TM4SF1 is highly expressed in breast cancer tissue. TM4SF1 may affect the occurrence, development and distant metastasis of breast cancer through various mechanisms. TM4SF1 may be a potential target for the treatment of breast cancer.

Objective:To assess the changes of myocardial biomechanical parametres of left ventricle of the patients with non-Hodgkin lymphoma(NHL) after anthracycline(ANTH) treatment by using two dimension speckle tracking imaging(2D-STI),and to study the value of 2D-STI in evaluating and monitoring the early cardiac dysfunction of the NHL patients induced by ANTH treatment. Methods:A total of 37 hospital patients who were firstly diagnosed as NHL (ANTH chemotherapy group) and 20 healthy volunteers (normal control group) were selected. The global longitudinal strain(GLS), global radial strain(GRS), global circumferential strain (GCS) and left ventricle twist(LVtw) of the subjects in two groups were detected before chemotherapy and 1,2,3 weeks after chemotherapy with 2D-STI; the GLS×LVtw was calculated. Results:There were no significant differences in the general clinical parameters of the subjects between normal control group and ANTH chemotherapy group before chemotherapy (P>0.05);the values of GLS, GRS and GCS of the patients in ANTH chemotherapy group after ANTH chemotherapy were decreased than those in normal control group and before ANTH chemotherapy(P<0.05),and GLS had the most obvious change. The GLS,GRS,GCS and GLS×LVtw of the patients after 3 cycles of chemotherapy in ANTH chemotherapy group were decreased (P>0.05).Four patients meet the diagnostic criteria of cardiac toxicity in the couse of chemotherapy.The sensitivity was 94%, the specificity was 66.7%, and the maximal Youden index was 0.667 when the △ GLS×LVtw=-64.53%×° was used as the cut-off point in predicting the occurrence of myocardial toxicity. Conclusion:2D-STI can detect the early changes of biomechanical parameters of left ventricular myocardium with highly sensitive in predicting early myocardial toxicity and early cardiac dysfunction caused by ANTH chemotherapy. It may be an effective way to predict the early myocardial toxicity of ANTH chemotherapy in the future.

Objective:To explore the differences of leukocyte subtypes and thyroid function of the patients with different thyrotoxicosis diseases, and to clarify the practical significance of leukocyte subtypes and thyroid function tests in the differential diagnosis of Graves disease thyrotoxicosisand destructive thyrotoxicosis. Methods:A total of 33 patients with Graves disease thyrotoxicosis and 30 patients with destructive thyrotoxicosis confirmed by clinical and laboratory examination were selected;the levels of neutrophils (Ne), lymphocyte (Ly), basophils (Ba), eosinophil (Eo) and mononuclear cells (Mo), serum free thyroxine (FT4), three free iodine thyroid former glycine (FT3),and thyroid stimulating hormone (TSH) of the patients in two groups were analyzed and the receiver operator characteristic curve (ROC) was used to evaluate the values of the indicators with statastical significance in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyrotoxicosis. Results:The levels of serum Eo,FT4 and FT3, and the modified data Eo/Mo, Eo×FT3/Mo of the patients in Graves disease thyrotoxicosis group were significantly higher than those in destructive thyrotoxicosis group(P<0.05);the levels of TSH and Mo of the patients were lower than those in destructive thyrotoxicosis group (P<0.05).The ROC curve analysis results showed that the sensitivities and specificities of Eo, Eo/Mo, Eo×FT3/Mo in the differential diagnosis of two diseases were good, and the best diagnostic boundaries were 1.54, 0.34, and 3.94. Conclusion:Eo, Mo, TSH, FT3, FT4, Eo/Mo, and EoxFT3/Mo could be regarded as the basis in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyrotoxicosis, and the practical significances of Eo, Eo/Mo, and Eo×FT3/Mo in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyotoxicosis are bigger.

Objective:To detect the expressions of Krüppel-like factor 12 (KLF12) protein in the cancer tissue and serum of the patients with colorectal cancer, and to investigate their clinical values. Methods:A total of 120 colorectal cancer patients were selected as case group, and 100 normal cases were regarded as control group. The cancer tissue and adjacent mucosa tissue were collected. Immunohistochemical method was used to detect the expressions of KLF12, nm23, CyclinE1, matrix metalloproteinase-2 (MMP-2), and MMP-9 proteins, and the relationships among these proteins were evaluated. ELISA assay was applied to detect the serum KLF12 levels of the patients in case group before operation and 1 month after operation the levels of serum KLF12 of the subjects in control group. Results:The positive expression rates of KLF12, CyclinE1, MMP-2 and MMP-9 proteins in cancer tissue were higher than those in adjacent mucosa tissue(χ²=66.155, 52.795, 64.515, 52.632; P<0.001), while the positive expression rate of nm23 protein in cancer tissue was lower than that in adjacent mucosa tissue(χ²=13.019, P<0.001). The Spearman analysis results showed that the expression of KLF12 protein was positively correlated with the expressions of MMP-2 and MMP-9 proteins (r=0.396 3, P<0.001; r=0.326 4, P<0.001); the expression of KLF12 was negatively correlated with the expression of nm23 protein (r=-0.227 3,P=0.013). The level of serum KLF12 of the patients in case group before operation was higher than that in control group. The cut-off value of ROC curve of serum KLF12 protein expression level was 3.795 μg·L^-1, and the area under the ROC curve was 0.834. The sensitivity and specificity were 60.8% and 94.0%, respectively. After operation, the serum level of KLF12 protein in the patients in case group was decreased, which was still higher than that in control group(t=2.708, P=0.007). The expression level of KLF12 protein in cancer tissue was positively correlated with the level of serum KLF12 protein(r=0.406 9,P<0.001). According to degree of lymph node metastasis, and the serum level of KLF12 protein in N3 group was the highest, and the lowest serum level of KLF12 protein was found in N0 group(F=21.731, P<0.001). Conclusion:KLF12 protein might be involved in the invasion and metastasis of colorectal cancer, and the detection of serum level of KLF12 protein could be valuable for the judgement of disease and prediction of lymph node metastasis of the patients.

Objective:To detect the levels of serum insulin-like growth factor 1 (IGF-1), insulin-like growth factor binding protein 3 (IGFBP-3) and the plasma proteinsof the patients with diabetic foot (DF), and to investigate the therapeutic effect of recombinant human growth hormone (rhGH) in the DFpatients. Methods:The DF patients (DF group,n=40), diabetes mellitus patients (DM group,n=40) and healthy adults (normal control group,n=40) were selected. The patients in DF group were divided into rhGH treatment group and conventional treatment group according to the treatment willingness(n=20).The patients in rhGH treatment group recevied 0.4 IU·kg^-1·d^-1 rhGH injection subcutaneously for 15 d, the interval between two courses was 5 d,with a total of 3 courses of treatment. The patients in convention treatment group received subcutaneous injection of the same dose of saline.The general clinical data before and after treatment, the serum IGF-1 and IGFBP-3 levels,the constitution of disease grading of patients, the levels of plasma TP and TRF, FBG, and the efficiency rate and wound healing time of the subjects in DF group and normal control group were detected. Results:The levels of serum IGF-1 and IGFBP-3 of the patients in DF group were significantly lower than those in DM group and normal control group (P<0.01); the serum levels of IGF-1 and IGFBP-3 of the patients in DM group were significantly lower than those in normal control group (P<0.05).After treatment, the serum levels of IGF-1, IGFBP-3 and plasma TP,TRF of the patients in rhGH treatment group were significantly increased compared with before treatment (P<0.05),and they were higher than those in conventional treatment group after treatment (P<0.05).There was no significant difference in the FBG levels of the subjects between DM group and normal control group(P>0.05).The effective rate of the patients in rhGH treatment group was higher than that in conventional treatment group (P<0.05). Conclusion:The efficiency of DF patients can be improved after treated with rhGH, and it is suitable for clinical promotion.

Objective:To compare the detection results of histopathology and MRI, CT of liposarcoma of extremities, and to provide guide for increasing the accuracy rate of clinical diagnosis. Methods:The data of 43 cases of liposarcoma of the extremities were collected.MRI plain scan and enhanced CT were used in 18 patients before operation. Sixteen cases were diagnosed by CT plain scan and enhanced scan before operation. Nine cases were diagnosed by MRI and CT plain scan and enhanced scan before operation. All the patients underwent the operation.The intraoperative pathological examination was performed in the lesion tissue.The diagnosed results were assessed according to the classification(high differentiated liposarcoma, mucoid liposarcoma, toliposarcoma and mixed liposarcoma). Results:In all the 43 cases,the upper limb lesion was found in 13 patients and the lower limb lesion was found in 30 patients. The average maximal diameter of the lesions was(14.6±6.1)cm. The fat cells were found in all the patients,with the pathological characteristics of adipocytes and lipopolysaccharide mixture exists,and accompanied by deep-stained pleomorphic cells;the CT examination results showed that the low density shadow with clear boundary and linear separated shadow after CT enhanced; the MRI image showed short signal intensity on T1 weighted imaging and long signal intensity on T2 long signal and short time reversal recovery sequence imaging and other signals; the mass existed the cord-like, linear shadow; the lesion surrounding nerve and blood vessels were squeezed, but the bone structure was relatively complete. The CT and MRI results of well-differentiated lipsarcome showed the fat signal, and the enhanced results showed the mild strengthening. The CT and MRI results of mucinous liposarcoma showed the cystic density and signal,and the enhanced results showed the clound-like orlatticed mild and moderate strengthening; the CT and MRI results of mixed liposarcoma showed the mixed imaging findings of above two types. Conclusion:The pathological examination can provide the exact basis for the diagnosis of type of liposarcoma; CT and MRI imagings can provide basis for judging the size of the tumor; MRI enhanced scan has an important role in judging tumor border.Combination of three kinds of detection methods can provide the basis for the diagnosis of the disease.

Objective:To make the fixed partial dentures with different materials and the magnetic resonance imaging(MRI)examination of head and neck was carried out to investigate the effects of fixed partial dentures on MRI. Methods:A qualified volunteer was recruited,who was conventionally restored using fixed partial denture. Co-Cr alloy, Au-Pt alloy and Zirconia were chosen as the inner crown materials,respectively. Three kinds of fixed partial dentures were put into the mouth of patient, and 2 kinds of MRI scan sequences (T1-TSE and T2-TSE)were performed. The scanning images without restorations were used as controls. The change of shape, image signal and involving layers of artifacts of different materials in three groups were observed.The maximum diameter of artifact of three materials in coronal,sagittal and axial images were detected.The safeties of fixed partial dentures were observed. Results:The artifacts were found in all the restorations and showed the changes of strengthen and weaken of the signal and the changs of shapes of images.The range of artifacts was the largest in the involved central plane and varied in sagittal, coronal and axial images.The number of involving layers of artifacts in Co-Cr alloy group was significantly more than those in Au-Pt alloy and Zirconia groups; the artifacts were limited in the repaired lateral maxillofacial region without the cervical spine and head region involvement. The artifacts in Co-Cr alloy group involved or slightly involved in all adjacent tissues, and the artifacts in Au-Pt alloy and Zirconia groups only slightly affected the mandibular teeth, lingual muscles and maxillary crowns. The maximum diameter of artifacts in Co-Cr alloy group was larger than those in Au-Pt alloy group and Zirconia group (P<0.05). No displacement was found in the prosthesis, and the patient had no discomfort. Conclusion:The artifact of the fixed partial denture can be seen in MRI examination. Co-Cr alloy or zirconia can significantly reduce the effect of artifact. Fixed adhesive prosthesis in mouth does not affect the safety of MRI examination.

Objective:To investigate the effect of Chinese traditional medicine-Shuanghuangbu combined with Xipayi Mouth Rinse on the periodontal indexes of the peri-implantitis patients,and to detect the curative effect of Shuanghuangbu combined with Xipayi Mouth Rinse in the treatment of peri-implantitis. Methods:A total of 40 patients diagnosed as peri-implantitis were randomly divided into control group and Chinese traditional medicine group(n=20).All of the patients received supragingival scaling and subgingival scaling, and then the peri-implant pocket of the patients in Chinese traditional medicine group were washed with Xipayi Mouth Rinse, while the fine probe slided slowly along the inner wall of peri-implant pocket into the bottom and retained the Shuanghuangbu,and retreated gently and made the medicine overflow the gingival margin.The patients in control group were given 3% hydrogen peroxide and 0.9% sodium chloride injection washed by turns. The indexes of covering modified plaque index(mPLI), probing pocket depth (PPD),modified sulcular bleeding index(mSBI) of the patients in two groups at baseline and after treatment were detected; Whatman#1 was applied to aspirate the peri-implant sulcus fluid (PISF)which were weighed at baseline and after treatment,and the level of interleukin-1β(IL-1β) in PISF was analyzed by ELISA method. Results:No differences were observed in age,sex,periodontal indexes, weights of PISF and IL-1β levels of the patients in two groups before treatment (P>0.05). Compared with before treatment,the indexes including PPD, mSBI,mPLI,weight of PISF and IL-1β level in PISF of the patients in two groups were decreased after treatment(P < 0.05).After treatment, the differences in mSBI, mPLI, weight of PISF and IL-1β level in PISF of the patients between control group and Chinese traditional medicine group were significant(P < 0.05).Compared with control group,the PPD level of the patients in Chinese traditional medicine group was decreased,but the difference was not significant(P > 0.05). Conclusion:The curative effect of Chinese traditional medicine-Shuanghuangbu combined with Xipayi Mouth Rinse is obsious,and the treatment can improve the periodontal status and reduce the level of IL-1β in PISF.

Objective:To evaluate the efficacy and safety of oxycodone combined with dexmedetomidine in the postoperative analgesia in the patients underwent laparoscopic radical surgery of colon cancer. Methods:Sixty colon cancer patients underwent laparoscopic radical surgery and recevied patient-controlled intravenous analgesia after operation were randomly divided into control group and experiment group(n=30) using The "Envelope Method".The patients in control group were given 0.5 mg·kg^-1 oxycodone, and the patients in experiment group were given 0.5 mg·kg^-1 oxycodone and 2 μg·kg^-1 dexmedetomidine. The parameters of analgesic pump in two groups were set as follows:the continuous perfusion rate was 1 mL·h^-1,the single bolus was 1.5 mL, the lock-out time was 10 min,and the total volume was 100 mL. The VAS scores of pain at rest and activity pain of the patients in two groups were evaluated at exiting the post anesthesia care unit (T₁), and at 6 h (T₂), 12 h (T₃), 24 h (T₄) and 48 h (T₅) after operation. The dose of analgesic drugs used, times of emergent rescue analgesia and anus exhaust time after operation of the patients in two groups were recorded; the postoperative adverse responses,vital signs and Ramsay scores of the patients in two groups were observed. Results:The VAS scores of activity pain at T₂-T₅ and pain at rest at T₂-T₄ of the patients in experiment group(P_T2-T5=0.000;P_T2=0.018,P_T3=0.001,P_T4=0.010) were lower than those in control group;the doses of analgesic drugs used and times of emergent rescue analgesia of the patients in experiment group were significantly lower than those in control group(P=0.000,P=0.029);the anus exhaust time after operation was significantly earlier than that in control group(P=0.030);the cases of nausea of the patients in experiment group were significantly lower than that in control group(P=0.023),and there were no significantly differences in the other various adverse effects of the patients between two groups(P>0.05). Conclusion:Oxycodone combined with dexmedetomidine is effective and safe for the postoperative analgesia after laparoscopic radical surgery of colon cancer.

Objective:To evaluate the efficacy of intravitreal lucentis or conbercept injection combined with ahmed glaucoma valve(AGV) implantation on the treatment of neovascular glaucoma(NVG). Methods:A total of 68 NVG patients(68 eyes) were divided into three groups according to the pretreatment (the intravitreal injection of lucentis or conbercept or no intravitreal injection 3-7 d before AGV implantation):lucentis group(n=26,given intravitreal injection of 10 g·L^-1 lucentis), conbercept group(n=21,given intravitreal injection of 10 g·L^-1 conbercept)and control group (n=21,given medicine or puncture of anterior chamber to smoothen the IOP and received AGV implantation). The panretina photocoagnlation(PRP) was performed in the patients with clear ocular media before intravitreal injection or AGV implantation. For the patients with cloudy ocular media,the intravitreal injection of anti-VEGF drugs was performed first and the PRP was performed when fundus could be seen. Results:Compared with before operation,the mean IOP and the number of antiglaucoma medications of the patients used in three groups after operation were significantly decreased(P < 0.01). There were no significant differences in the successful rates of operation of the patients in three groups at different time points after operation(P>0.05). 1 month and 3 months after operation,the BCVA of the patients in lucentis and conbercept groups was significantly better than that in control group(P<0.05).There were no significant differences in the incidence of complications of the pateints between lucentis and conbercept groups at different time points after operation (P>0.05). Conclusion:Intravitreal injection of anti-VEGF drugs combined with AGV implantation is effective and safe in the treatment of NVG. In short term, it can improve the vision, relieve the pain and reduce the complications.

Objective:To evaluate the clinical effect of pereutaneous vertbroplasty(PVP) combined with implantation of iodine-125(¹²⁵I)radioactive particle in the treatment of vertebral metastasis,and to provide basis for the treatment of vertebral metastasis. Methods:A total of 69 patients with vertebral metastasis were divided into test group(n=32) and control group(n=37); the patients in test group were treated with PVP comined with implantation ¹²⁵I radioactive particle and the patients in control group were treated with PVP only. The heights of anterior and posterior vertebral bodies of the patients before and after treatment were detected by X-ray. The numerical rating scale(NRS)scores, pain relief rate and the incidence of surgical complications of the patients were recorded before operation and 1 d, 1 week, 1 month, 3 months, and 6 months after operation. Results:The operation was successfully performed in all the patients without local bleeding; there were no movement dysfunction and nerve compression phenomenon.There was no leakage of bone cement. All the ¹²⁵I radioactive particles located well and there was no particle obscission. The heights of vertebral bodies of the patients in two groups after operation were increased compared with before operation(P<0.05).The NRS scores of the patients in two groups s at 1 d, 1 week, 1 month, 3 months, 6 months after operation were significantly decreased compared with before operation(P<0.05);compared with control group,the NRS scores of the patients in test group at 1 d,1 week,1 month,3 months,6 months after operation were decreased(P<0.05).The incidence of pulmonary embolism or radiation myelitis complications was about 4.3% in 69 patients. Compared with control group, the difference in the incidence of complications of the patients in test group was not significant(P<0.05). Conclusion:PVP combined with ¹²⁵I radioactive particle implantation is a safe and effective method in the treatment of vertebral metastasis, which can relieve the pain of the patients obviously compared with PVP.

Objective:To observe the process and curative effect of12 cases of recurrent patellar dislocation treated by double bundle medial patellofemoral ligament (MPFL) reconstruction with anterior half peroneus longus and lateral retinacular release, and to investigate the etiology and treatment method of recurrent patellar dislocation. Methods:A total of 12 patients with recurrent patellar dislocation were enrolled in this study, the MPFL was reconstructed with the anterior half peroneus longus, patellar fixation with suture anchors was completed with 2 parallel 5.0 mm anchors which were spaced 1.0 cm apart at the anatomic insertion site of the native MPFL,the femoral side was secured with a interference screw, and the lateral retinaculum was released at the same time.The Lysholm score,IKDC score,congruence angle,J sign,grind test,and apprehension test of the patients before and after operation were detected. Results:The mean follow-up period was 16.4 months,and the Lysholm score of the patients before operation was lower than the last follow-up(t=9.03, P < 0.001); the IKDC score of the patients before operation was lower than the last follow-up (t=9.75, P < 0.001); the congruence angle of the patients before operation was larger than after operation (t=7.22,P < 0.001). All of the patients demonstrated the positive results before operation in Jsign,grind test,and apprehension test, and the negative results in J sign,grind test,and apprehension test after operation. No patient appeared pateela fracture and recurrence of patellar dislocation during the follow-up period. Conclusion:The curative effect of reconstruction of the MPFL with anterior half peroneus longus combined with lateral retinacular release is well in the treatment of recurrent patellar dislocation, which is suitable for clinical promotion.

Objective:To fabricate an all-ceramic crown which copies the morphology of preoperative tooth in one patient with only small occlusal dentin defects with completely digital design(CDD) technology, and to find a more precise and feasible solution to fix the tooth detect. Methods:The tooth treated with root canal therapy had the complete axial surface and small occlusal dentin defects. In order to keep the morphology of preoperative tooth, the digitized crown was designed with the information of preoperative tooth by oral scanning with CS3500 and using the "Biogeneric Copy" function of exocad design software, then the digitized information of the teeth was collected and transformed and the simulate zirconia crown was mode by 3D colloidal deposition technique. The patient's comfort and satisfaction were investigated by observing the morphology, the interproximal contacts, and the occlusion of crown. Results:The morphology of definitive restoration was similar to the preoperative tooth, which presented good interproximal contact and occlusion, and the patient was greatly satisfied without discomfort and foreign body sensation. Conclusion:A more accurate and personalized restoration approach can be implemented with the application of CDD to fabricate new ceramic crowns which copy the preoperative tooth.

Objective:To analyze the clinical data of 2 patients with primary intracranial chondrosarcoma, and to improve its climical awareness and management. Methods:One patient admitted to the hospital because of his left eye with oculomotor palsy,and he was considered as craniopharyngioma before operation and received radiotherapy after operation. The other patient admitted to the hospital for the vision loss,and he was considered as chordoma and received subtotal resection. Results:Both two patients were confirmed as chondrosarcoma by postoperative pathological examination. One patient's clinical symptoms relieved after operation, and there was no obvious progress from the begining of follow-up to now; the other patient died of severe intracranial infection on the 10th day after operation. Conclusion:Primary intracranial chondrosarcoma is easy to be misdiagnosed before operation and needs to be confirmed by postoperative pathology. Operation is the first choice for its treatment, and the postoperative radiotherapy can achieve the good results.

Objective:To discuss the pathogenesis, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of the patient with rituximab (RTX)-induced interstitial lung disease(ILD)(RTX-ILD). Methods:A male patient aged 19 years old was confirmed as classical Hodgkin's lymphoma more than 2 years ago, and received autologous hemopoietic stem cell transplantation(HSCT) when the disease recurred 1 year ago. Two months after HSCT, the maintenance chemotherapy was given every month with 375 mg·m^-2 RTX, but the mediastinal recurrence appeared during the period, then the mediastinal residual lesion radiotherapy was done for 10 times. However, 8 d after the third RTX chemotherapy, the patient experienced post-exercising tachypnea, fever, cough, chest congestion, and limb weakness. The chest CT scanning results suggested ground gloss opacity on both lungs, and blood gas analysis suggested a hypoxemia. Neither antibacterial nor antifungal treatment worked well. Afterwards, the sequential etiological examination showed the negative results, and the bronchoscope examination showed the normal results as well. Considering about the possibility of RTX-ILD, RTX was stopped immediately and turned to intravenous infusion of methylprednisolone 40 mg per day. Results:Five d later, the symptoms of the patients were improved, and the follow-up methylprednisolone was changed to oral and gradually reduced to discontinuation. Conclusion:RTX-ILD is relatively rare,and its clinical manifestations lack specificity. Hormonotherapy is the main method in treatment of RTX-ILD. The patients sensitive to hormonotherapy have better prognosis than those insensitive to hormonotherapy.

Objective:To investigate the diagnosis and prognosis of one patient with carcinoma ex pleomorphic adenoma (CXPA) composed of three malignant components in parotid gland, and to raise the clinicians' awareness of the disease. Methods:A patient was presented to hospital because of the mass in left parotid gland region for more than 30 years and the accompanied pain lasted for one month.After color ultrasonography,the left parotid gland tumor resection was performed. Results:The operation was successful.The postoperative pathological diagnosis results comfirmed as CXPA, it was composed of three malignant components, including non-specific adenocarcinoma, ductal carcinoma, and myoepithelial carcinoma. Conclusion:CXPA composed of three malignant components at the same time are extremely rare.CXPA is difficult to diagnose and its prognosis is poor.The clinicians are supposed to improve the understanding of CXPA.

Objective:To investigate the clinical application value of ultrasound in the diagnosis and differential diagnosis of Takayasu arteritis (TA). Methods:A total of 39 suspected TA patients were examined by ultrasonography, erythrocyte sedimentation rate(ESR), C reactive protein (CRP)and CT angiography(CTA), and the ultrasound examination results were compared and analyzed with the final chinical diagnosis results. Results:Of the 39 cases of suspected TA patients which had been examined by ultrasound, 33 cases were diagnosed as TA, 2 cases were diagnosed as atherosclerosis(AS), 2 cases were diagnosed as fibromuscular dysplasia (FMD) and 2 cases were diagnosed as thromboangiitis obliterans(TAO). Compared with final chinical diagnosis results, 1 FMD patient was misdiagnosed as TA, and the rest of the diagnostic results were consistent with the final chinical diagnosis results. Conclusion:Ultrasound examination can provide objective and accurate basis for the diagnosis and differential diagnosis of TA, and it is an effective method of imaging examination.

Objective:To explore the purification technology parameters for polyphenols from vitis amurensis seed extract with AB-8 macroporous resin and to establish the optimum purification process conditions, and to provide technical reference for further development and utilization of vitis amurensis seeds. Methods:The vitis amurensis seeds were regarded as raw material, the concentration of vitis amurensis seed polyphenols was detected,and the adsorption rate and desorption rate of polyphenols were calculated. The AB-8 macroporous resin was used to purify the polyphenols from vitis amurensis seed extract,and the purification technology parameters of vitis amurensis seed polyphenols were optimized by absorption-desorption experiment.The concentrations of polyphenol from vitis amurensis seed extract were compared between before and after purification,and the effect of the purification process was verified. Results:The AB-8 macroporous resin had good absorption and desorption properties to crude the extract of vitis amurensis seed polyphenols,and the adsorption rate and desorption rate were 90.48% and 71.42%.The optimum adsorption conditions and desorption conditions were as follows:the sample concentration was 10 g·L^-1, the sample flow rate was 3 mL·min^-1, the eluting concentration was 60%,the elution velocity was 2 mL·min^-1,and the eluting volume was 2-fold column volume. Under this condition,the concentration of polyphenols from vitis amurensis seed extract after purification was increased from 35.02% to 88.8%. Conclusion:AB-8 macroporous resin has better purification effect,and it is worthy of popularization and application.