Haemophilia is caused by lack of coagulation factor VIII or IX in patients′blood with inadequate hemostasis.Currently recombinant coagulation factor VII(rFVII)produced in different cells is used against clini-cal bleeding of haemophilia patients.To enhance the production and activity of rFVII;some eukaryotic cells such as baby hamster kidney(BHK);Chinese hamster ovary(CHO);insect cell and fish embryo;were used to express rFVII.Meanwhile;the effect of functional gene on the activity of rFVII and the limitation of rFVII production caused by post-translational modification were investigated by different methods.The role of rFVII in hemostasis;synthesis of rFVII in different eukaryotic cells and impact on production of post-translational modification are reviewed in this article.

Objective To explore the preparation of liposomal clodronate and investigate its inducing effects on the apoptosis of peritoneal macrophages in rats after severe acute pancreatitis (SAP).Methods Liposomal clodronate was prepared by means of thin film. SAP rat model was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. The peritoneal macrophages were obtained from SAP rats. After exposure to different doses of liposomal clodronate (50, 100,150 μl), the PM proliferation was determined by MTT colourimetry. The apoptosis of PM was measured by flow cytometry and agarose gel electrophoresis, respectively. Results The prepared liposomal clodronate had a suitable encapsulation efficiency of clodronate (5.8%) with an average size of 200 nm. The spherical shape of liposome was confirmed by transmission electron microscope. Exposed to liposomal clodronate of different doses resulted in a obvious growth depression (P＜0.01). The apoptotic rate of the PM was (10.32±0.34) %, (18.16±0.49)% and (29.87±0.35)% in three different dose groups and the difference was marked (P＜0.01). 1.2% of agarose gel electrophoresis of DNA extracted from apoptotic macrophages induced by liposomal clodronate showed clearer and characteristic ladder following the liposomal clodronate concentration. Conclusion Liposomal clodronate has a definite effect on peritoneal macrophages in SAP rats.

Objective To establish a model of cervical spondylosis in rats. Methods Sixty SD rats (four months old) were ran-domly divided into control, muscle imbalance and posterior column instability group, ten rats in each group. Then the degeneration of X-ray film and motion function were evaluated by oblique board test. Results After analyzed the cervical films of control groups, the nature curve still existed, vertebral clearance had no abnormal. No ostensis spurs or ossifies in Luschka joint and joint processes were found. But in mus-cle imbalance and posterior column instability groups, the nature curve was disappeared or stiff, vertebral clearance were stenosis, osteosis spur formed, ossify or sclerosis emerged in Lnschka joint and joint process. Compared with control group in oblique board test, the muscle force decreased in muscle imbalance and posterior column instability group. There were no statistic significant difference between 2 and 4 months of control group, and also no difference in muscle imbalance and posterior column instability group, but there were significant differ-ence in 2 and 4 months of muscle imbalance and posterior column group. Conclusion Not only cervical posterior column instability but also muscle imbalance could result in cervical spendylosis, both muscle force balance and posterior column were important factors in maintaining spine stability.

Objective To investigate the protective effects of lipsomal clodronate on renal injury in rats with severe acute pancreatifis and the assessment of renal injury. Method Totally 48 rats were randomly divided into three group:normal control group (C);SAP group, in which rats were treated with pure liposomal (P);treatment group, in which SAP rats were treated with liposomal clodronate disodium(T). The SAP model of rat was induced by injection of 5 % sodium taurochohte beneath the pancreatic membrane. Rats of normal control group received isovolumetric injections of 0.9% physiological saline solution instead of sodium taurocholate. Blood samples were collected to measure AMS,BUN,Cr,IL-6 and IL-12 at 2 hors, 6 hours after SAP. At the same time, the samples of pancreatic and renal tissues were taken for observing the pathological changes. Results Compared with controlgroup, serious renal and pancreatic damages were found in group P, and the AMS, BUN, Cr levels elevated signifi-candy (P < 0.01). Compared with group P,the renal and pancreatic damages were attenuated in group T, and the levels of Cr and AMS decreased significantly (P < 0.01), and the IL-6, IL-12 were decreased at 2 hours and 6 hours (P < 0.01). The BUN decreased significantly at6 hours (P < 0.05). Conciusions Excessive release of inflammatory mediator play an important role in renal injury in SAP. Lipsomal clodronate disodium can alleviate the damage of pancreas and kidney.

Objective To investigate the protective effect of lipsomal clodronate against hepatic injury in rats with acute necrotizing pancreatitis (ANP). Methods 48 SD rats were randomly divided into control group, ANP group and lipsomal clodronate group, respectively. The models of ANP were established by injection of sodium taurocholate into the pancreatic capsule. Lipsomal clodronate was prepared by means of thin film. Blank liposomes and clodronate-containing liposomes was injected via caudal vein in ANP group and lipsomal clodronate group, respectively. The rats were sacrificed at 2, 6 h after ANP induction, the serum levels of ALT, AST and AMS, IL-6,IL-12 were measured, and pathologic changes of liver and pancreas were observed. Results At 6 h, serum level of ALT was (73 ± 11) U/L, (257 ± 33) U/L and (184 ± 29) U/L in control group, ANP group and lipsomal clodronate group, respectively;serum levels of AST were (190 ± 32)U/L, (590 ± 70)U/L and (430±52)U/L, respectively;serum levels of AMS were (814±80)U/L, (5031 ± 471) U/L and (2843 ± 236) U/L, respectively, serum levels of IL-6 were (26.7 ± 5.7) pmol/L, (218.0 ±4.7)pmol/L and (112.3 ± 8. O) pmol/L, respectively;serum levels of IL-12 were (4. 2 ± 1.0) pmol/L,(309.5 ± 8.5) pmol/L and (153.7 ± 6.3) pmol/L. The values in ANP group and lipsomal clodronate group were significantly higher than those in control group, while the values in lipsomal clodronate group were significantly lower than those in ANP group (P < 0. 01). Pathologic changes of liver and pancreas were significantly attenuated in lipsomal clodronate group. Conclusions Intravenous liposomal clodronate could exert protective effects on the hepatic injury in rats with ANP.

Objective To investigate the apoptosis of Kupffer cell (KC) induced by lipsomal clodronate in rat with acute necrotizing pancreatitis (ANP). Methods Lipsomal clodronate was prepared by means of thin film, the model of ANP was established by injection of 5% sodium taurocholate of 4 ml/kg into the pancreatic capsule. The Kupffer cells were obtained from ANP rat. After exposure to different doses of lipsomal clodronate (0, 50, 100, 150 μl) , then the proliferation and apoptosis of KC was measured by MTT, flow cytometry and agarose gel electrophoresis of DNA. Results The prepared lipsomal clodronate had an average size of 100～200 nm, the spherical shape of liposome was uniform and confirmed by transmission electron microscope. When exposed to different concentration of lipsomal clodronate for 24 h, the growth suppression rate was 17. 4% , 24. 2% and 31. 1% , respectively, while the apoptosis rate of the KC was (14. 12 ±0.37)% , (18.74±0.43)% and (27.51 ±0.39)%, respectively; the difference was statistically significantly (P<0. 01) , the DNA of KC began degradation and gradually showed clear and characteristic ladder. Conclusions Lipsomal clodronate could induce apoptosis and suppress the growth of Kupffer cells in ANP rats.

Objective To investigate the protective effect of clodronate SPIO liposomes on liver injury of rats with severe acute pancreatitis(SAP)and the role of MRI in evaluating the extent of liver injury.Methods Superparamagnetic Fe3O4 nanoparticles were prepared by chemical coprecipitation.Clodronate-SPIO-containing liposomes was prepared by the thin-film method.SAP models were prepared by a uniform injection of sodium taurocholate(2 ml/kg body weight)into the subcapsular space of the pancreas.SD rats were randomly divided into control group,SAP plus SPIO group, and clodronate-SPIO-containing liposome group.Six hours after SAP models were available,T2-weighted MRI scanning(in the same plane)of the liver of rats in each group were performed.At the end of the scanning,blood samples were taken from the supcrior mesenteric vein to measure the contents of serum ALT and AST.Meanwhile, The pathological changes in the liver and pancreas were observed.Results Transmission electron microscopic examination showed that liposomes had a uniform size.No changes in the pancreas of rats in control group were noted.The pathological changes in the pancreas and liver of rats in SAP plus clodronate-SPIO-containing liposome group were significantly milder than those in SAP plus SPIO liposome group.The contents of serum ALT and AST in rats in SAP plus SPIO liposome group were significantly higher than those in control group(P＜0.01), while the contents of serum ALT and AST in rats in SAP plus clodronate-SPIO-containing group were significantly lower than those in SAP plus SPIO liposome group(P＜0.01).The MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus clodronate-SPIO-containing liposome group was significantly lower than that in control group.The significant changes in the MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus Clodronate-SPIO liposome group were noted(P＜0.01).Conclusion Clodronate-containing liposomes have protective effects against liver injury in SAP rats and SPIO can be used as a tracer for MRI examination.

This study was focus on investigating the anti-liver fibrosis effects of insulin-like growth factor-1 (IGF-1) in vitro.The effects of IGF-1 on human liver L-02 cell viability and cell cycle were observed.CC14-induced L-02 cell injury was set up to detect the anti-apoptotic activity of insulin-like growth factor-1 (IGF-1).Transforming growth factor β1 (TGF-β1) induced hepatic stellate cell line (HSC-T6) were used as a liver fibrosis model in vitro to analyze the effects of IGF-1 on the expression of liver fibrosis proteins and intracellular TGF-β1/Smad signaling pathway in HSC-T6 cells.The results showed that IGF-1 could relieve the growth inhibition effects of TGF-β1 on L-02 cells,increase the viability of L-02 cells injured by CCl4,decrease the expression of liver fibrosis proteins,and inhibit the TGF-β1/Smad signaling pathway by inhibiting the phosphorylation of Smad3.Our study suggested that IGF-1 exerted anti-liver fibrosis effects by stimulating L-02 cells proliferation,reducing cell damage and inhibiting ECM accumulation via interfering TGF-β1/Smad signaling pathway.

This paper introduces the preamplifier for the microcomputer-detecting system of 3-Channel ECG signals and its technical specifications. The practical experiments and clinical applications show that its performances conforms with all the requirements of the system.
Amplifiers, Electronic ; Electrocardiography ; instrumentation ; Equipment Design ; Heart Diseases ; diagnosis ; Humans ; Image Enhancement ; instrumentation ; Microcomputers ; Signal Processing, Computer-Assisted

OBJECTIVE: To observe the effect of warm needle moxibustion (WNM) on knee-joint swelling and expression of sirtuin 1 (SIRT 1, a class Ⅲ histone/protein deacetylase) and nuclear factor κB (NF-κB) in synovial tissue in rats with rheumatoid arthritis (RA). METHODS: Fifty SD rats were randomly divided into five groups:normal control, model, SIRT 1-inhibitor, WNM, and SIRT 1-inhibitor plus WNM (inhibitor+WNM), n=10 rats in each group. The RA model was established by subcutaneous injection of bovine type Ⅱ collagen at the sole, once daily for 21 days. The WNM was applied to bilateral "Zusanli" (ST 36), "Shenshu" (BL 23) and "Xuanzhong" (GB 39) for 15 min, once daily for 21 days, beginning on the 1st day of modeling. For rats of SIRT 1-inhibitor group and inhibitor+WNM group, nicotinamide solution (10 mmol·kg-1·d-1) was injected intraperitoneally on the 1st, 7th, 14th and 21th days of modeling. The swelling volume of the affected knee-joint was measured by water drainage method. The contents of IL-1 β, IL-6 and IL-8 in the serum were measured by radioimmunoassay. The expression of SIRT 1 and NF-κB p 65 proteins in the synovial tissue of knee-joint were detected by immunoblotting. RESULTS: After modeling, the knee-joint volume, serum IL-1 β, IL-6 and IL-8 contents, and the expression of NF-κB p 65 protein in synovial tissue of knee-joint were considerably increased relevant to the normal control group (P<0.01), while the expression of synovial SIRT 1 protein was significantly down-regulated (P<0.01). Following WNM intervention, the volume of the knee-joint, serum IL-1 β, IL-6 and IL-8 contents and synovial NF-κB p 65 expression were significantly decreased relevant to the model group (P<0.05), and the down-regulated expression of SIRT 1 protein was notably suppressed (P<0.05). No significant difference was found between the WNM and inhibitor+WNM groups in the knee-joint swelling degree (P<0.05). The contents of serum IL-1 β, IL-6 and IL-8, and the expression of synovial NF-κB p 65 protein of the WNM group were significantly lower than those in the inhibitor+WNM group (P<0.05), while the expression level of SIRT 1 protein was evidently higher than that in the inhibitor+WNM group (P<0.05), suggesting the effects of WNM in down-regulating serum inflammatory cytokines and NF-κB p 65 expression, and in up-regulating SIRT 1 expression after administration of SIRT 1 inhibitor. CONCLUSIONS: Warm needle moxibustion can relieve inflammatory reactions of RA rats, which may be associated to its effect in modulating SIRT 1/NF-κB p 65 signaling in the synovial tissue.