Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

BACKGROUND:Staphylococcal infections and its biofilm formation can occur when orthopedic implants or wound is healing, and are regulated by bacterial population sensing mechanism. RNAIII inhibiting peptide intervenes the quorum-sensing system of staphylococcal and blocks the signal transduction among staphylococcal cells, and inhibits staphylococcal biofilm formation, and then prevents staphylococcal infections.
OBJECTIVE:To investigate the influence of RNAIII inhibiting peptide on the adhesion of staphylococcus epidermis to the Hela cells.
METHODS:The Hela cells were cultured in vitro. There were four groups in this study. In the blank group, saline with dimethyl sulfoxide was added in each wel . In the RNAIII inhibiting peptide group, dimethyl sulfoxide solution containing RNAIII inhibiting peptide was added. In the levofloxacin group, levofloxacin was added. In the combination group, the dose was in accordance with above methods. Using intergroup control method, the adhesion of staphylococcus epidermis to the Hela cells was compared under the effects of saline, RNAIII inhibiting peptide and levofloxacin and their combination.
RESULTS AND CONCLUSION:In the blank group, abundant bacterial adhered to Hela cells. The number of adhered bacteria was significantly lower in each medicine group than in the blank group (P<0.001). The spot count was significantly lower in the levofloxacin group than in the RNAIII inhibiting peptide group (P<0.05). In the combination group, the number of bacteria adhered to Hela cells was decreased (P<0.01). Results verified that RNAIII inhibiting peptide effectively suppressed the adhesion of staphylococcus epidermis to the host cells, and showed synergistic effects on antibiotics.

BACKGROUND: A new material of porous carbonated hydroxyapatite cement (PCHC) is discovered using foaming technique.The new material characterizes original solidification and forms porous structure.OBJECTIVE: To investigate the biomechanical effect of PCHC on repairing cancellous bone defect.METHODS: Among 30 New Land rabbits, 25 ones were considered as surgery group, whose bilateral condyles of femur was used to establish bone defect model (5.5 mm diameter and 12 mm depth). PCHC was implanted into the left side, which was considered as the experimental group, and carbonated hydroxyapatite cement (CHC) was implanted into the right side, which was considered as the control group. Another 5 rabbits were used as normal mechanical control group. Both PCHC and CHC were dip in simulated body fluid (SBF) to test mechanical intension. PCHC and CHC were then implanted into muscles of back in the surgery group. Rabbits Were-sacrificed after 2, 4, 8, 12, and 16 weeks postoperatively. Mechanical analysis was tested following intra-bone and intramuscular implantation, and compressive strength was then tested following dipping into SBF.RESULTS AND CONCLUSION: PCHC: Intra-bone mechanical strength was lower at 2 weeks, the lowest at 4 weeks, but then closed to intension of normal cancellated bone at 8 weeks, higher than normal cancellated bone at 12 weeks, and recovered to the level of normal cancellated bone at 16 weeks. CHC: Intra-bone strength was higher than that of PCHC at 2 weeks, decreased at 4 weeks, gradually increased at 8, 12, and 16 weeks, but still lower than intension of normal cancallated bone. Compressive strength of both PCHC and CHC was not changed following dipping in SBF; however, compressive strength was changed remarkably following intramuscular implantation. The results demonstrated that PCHC characterized by immobilization in situ and mechanical supporting. Thus it could be used for one kind of bone substitute material to repair the bone defect.

OBJECTIVE: To evaluate the histocompatibility of poly (lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide (PLGNRIP) sustained release microspheres.METHODS: The crude peptide comprising N to C-terminals was synthesized using Fmoc method. The crude synthetic RNAⅢ peptide was purified by reverse phase high performance liquid chromatography, followed by component harvesting according to ultraviolet absorption peak, and freeze-drying. PLGNRIP sustained release microspheres with a diameter of 50-70 pm were prepared using liquid-phase multiple emulsion method. The histocompatibility of PLGNRIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test, MTT cytotoxicity test, intramuscular implantation test, sensitivity test, and pyrogen test.RESULTS: Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass. MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%, with cytotoxicity grade 1, which indicated no cytotoxicity. Intramuscular implantation tests showed that at 4 weeks after implantation of RiP powder or PLGNRIP microscopes, no obviously congested, degenerated, or necrotic tissue was observed. All RIP powder and a part PLGNRIP microscopes were degraded. Fibroblasts accounted for a large proportion in all cells. NO inflammatory cell infiltration, involving neutrophits and multinucleated giant celts, was observed. Sensitivity test rasults displayed that the average primary irritation index was 0.38, 0.33, arid 0.31 in the eluent stock solution, 2% dinitoflruorobenzene, and physiological saline-administerd groups, respectively. Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5 ℃ and the sum of fervescence was under 1.3 ℃ .This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION: PLGNRIP sustained release microspheras exhibit good histocompatibility.

Objective To investigate the influences of survivin down-regulation on cell G2/M phase arrest,apeptosis and sensitivity to carbon ion irradiation. Methods Small interfering RNA (siRNA) targeting survivin mRNA was designed, in vitro chemo-synthesized and transfected into SMMC-7721 cells. Survivin mRNA expression in SMMC-7721 cells was measured by real-time PCR, and the apeptotic rates by Annexin-FTTC at 24 and 48 h after transfection. Cell G2/M phase arrest after transfection was assessed with flow eytometry as well. Cellular sensitivity to high-LET carbon ions was determined by means of colony-forming assay. Results The expressions of survivin at mRNA level were down-regulated to be 59% and 39% in relation to the non-treated cells at 24 and 48 h after siRNA transfeetion, respectively. G2/M phase arrest in SMMC-7721 cells at 24 h after transfection was observed while much more obvious at 48 h. The apeptotic rate of SMMC-7721 cells was 21.41 % at48 h after survivin siRNA transfection, which was significantly higher than that of the cells transfected with negative siRNA. Moreover, a decreased clonogenic survival in siRNA treated group was shown. Conclusion Down-regulation of survivin gene expression in SMMC-7721 cells by siRNA could effectively induce cell apeptosis and G2/M phase arrest, and enhance the cellular radiosensitivity to high-LET heavy ions.

OBJECTIVE: To evaluate blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide (PLGA/RIP) delayed release microspheres.METHODS: ① Preparation of PLGA/RIP microspheres: The solid-phase synthesis (Fmoc) method was used to synthesize RIP crude sample from C end to N end; the synthesized crude peptide was purified by the reverse phase high performance liquid chromatography. According to UV absorption peak, the components were collected and freeze-dried, to obtain RIP purifications. Then liquid-phase multiple emulsion method was used to prepare PLGA/RIP microspheres at the diameter of 50-70 μm. ② Preparation of eluent: The PLGA/RIP microsphere powders were eluted with sterile physiological saline at 37 ℃, to prepare 1 g/L eluent; then 0.5 g/L eluent was obtained adding equal volume of sterile physiological saline. The hemolysis test, blood clotting test, and platelet aggregation test were conducted to measure prothrombin time and activated partial thromboplastin time, to observe the influence on rabbit leucocytes, erythrocytes and thrombocytes, and to preliminarily evaluate the blood compatibility of PLGA/RIP microspheres. RESULTS: ①The haemolysis rates of eluent stock solution and 0.5 g/L eluents were 3.24% and 2.67% respectively, which were in coincidence with the criteria of medical biomaterials, less than 5%. ② The eluent stock solution and 0.5 g/L eluents of PLGA/RIP microspheres had no significant effect on rabbit clotting time, prothrombin time and activated partial thromboplastin time, the number of rabbit leucocytes, erythrocytes and thrombocytes, as well as platelet aggregation.CONCLUSION: PLGA/RIP delayed release microspheres have a good blood compatibility.

AIM: RNAⅢ inhibiting peptide (RIP) has been previously proved to possess good histocompatibility and safety for preventing and curing staphylococcal infection, and this study evaluated histocompatibility of polyaiticglycolic acid/RIP (PLGA/RIP) sustained release microsphere. METHODS: The experiment was performed in the Orthopedic Institute, Pharmacologic Research Institute and Animal Experimental Center of General Hospital of Chinese PLA from October 2005 to October 2007.①Preparation of PLGA/RIP: The solid-phase synthesis (Fmoc) method was used to synthesize RIP from C end to N end, then the synthesized peptide was purified by the reverse phase high performance liquid chromatography, and composition was collected by means of ultraviolet absorption peak. The purified RIP was obtained after freezing and drying. Liquid-phase multiple emulsion method was used to synthesize PLGA/RIP microsphere of 50-70 ?m diameter.②Acute general toxicity test was studied in PLGA/RIP. Effect of PLGA/RIP on the cell proliferation was detected with cytotoxicity test by MTT method. Intramuscular implanting test was used to observe the irritation reaction of muscles by implantation materials. Sensitivity test was used to observe the sensitization of PLGA/RIP. Changes of animal's body temperature were determined with pyrogen test. RESULTS: ①Acute general toxicity test: Neither toxicosis reaction nor animal death was found after animals were injected with 100% and 50% eluents of PLGA/RIP peritoneally. Animal's body weight was not changed significantly.②Cytotoxiciity test by MTT method: The average proliferation rate of cell in two kinds of eluents exceeded 85% and cytotoxicity was graded in 1 rank, indicating no cytotoxicity.③Intramuscular implanting test: At 4 weeks after RIP and PLGA/RIP were implanted into the animals, there was not obvious synathresis, denaturation or necrosis in tissues. No inflammatory cell infiltration occurred around the materials. There had been the fibrous capsules around the materials.④Sensitivity test: Average primary irritation index of three groups were 0.38, 0.33 and 0.31 respectively. There was no significant difference among three groups.⑤Pyrogen test: Fervescence of each animal in the experiment was under 0.5 ℃, confirming that the materials had no pyrogenic characteristics. This was in coincidence with evaluation criterion of pyrogen test. CONCLUSION: PLGA/RIP has good histocompatibility and safety, without general toxic reaction, cytotoxicity, immunological rejection, hypersensitive response or pyrogenic characteristics.

OBJECTIVE:To evaluate the histocompatibility of poly(lactic-co-glycolic acid)/RNA Ⅲ inhibiting peptide(PLGA/RIP)sustained release microspheres.METHODS:The crude peptide comprising N to C-terminals was synthesized using Fmoc method.The crude synthetic RNAIII peptide was purified by reverse phase high performance liquid chromatography,followed by component harvesting according to ultraviolet absorption peak,and freeze-drying.PLGA/RIP sustained release microspheres with a diameter of 50-70?m were prepared using liquid-phase multiple emulsion method.The histocompatibility of PLGA/RIP sustained release microscopes were preliminarily evaluated through the use of acute general toxicity test,MTT cytotoxicity test,intramuscular implantation test,sensitivity test,and pyrogen test.RESULTS:Acute general toxicity test results showed that all included animals survived and presented with no toxicosis reaction and obviously changed body mass.MTT cytotoxicity test results revealed that the average relative growth rate of cells from two eluents was over 85%,with cytotoxicity grade 1,which indicates no cytotoxicity.Intramuscular implantation tests showed that at 4 weeks after implantation of RIP powder or PLGA/RIP microscopes,no obviously congested,degenerated,or necrotic tissue was observed.All RIP powder and a part PLGA/RIP microscopes were degraded.Fibroblasts accounted for a large proportion in all cells.No inflammatory cell infiltration,involving neutrophils and multinucleated giant cells,was observed.Sensitivity test results displayed that the average primary irritation index was 0.38,0.33,and 0.31 in the eluent stock solution,2% dinitoflruorobenzene,and physiological saline-administerd groups,respectively.Pyrogen test results showed that fervescence of each rabbit in the experiment was under 0.5℃ and the sum of fervescence was under 1.3℃.This is in coincidence with evaluation criteria of pyrogen test.CONCLUSION:PLGA/RIP sustained release microspheres exhibit good histocompatibility.

OBJECTIVE To investigate the in vitro effect of 4 kinds of implant materials on the biofilm formation of Staphylococcus epidermidis.METHODS S.epidermidis was cultured,purified and identified.Susceptibility test was done for S.epidermidis and the ability to produce biofilm was proven.The test samples were made into wafer shape for titanium alloy,Co-Cr-Mo alloy,ultrahigh molecular weight polyethylene(UHMWPE)and home-made polymethyl methacrylate(PMMA).S.epidermidis was cultured with 4 kinds of test samples for 5 days respectively.Bacteria adhering on surfaces of 4 kinds of test samples were dissolved with trypsin,and then diluted into bacterial suspensions.Each bacterial suspension was inoculated quantitatively and CFU were counted.Biofilm on surfaces of 4 kinds of test samples prepared by vacuum drying method was observed with SEM.RESULTS The strain proven to be S.epidermidis,was resistant to semisynthetic penicillins,and could produce biofilm.CFU count showed that CFU were the most on the UHMWPE surface and the number of CFU were(24.96?1.459)?105.CFU were(17.44?1.883)?105 on the PMMP surface.(0.424?0.065)?105 CFU were discovered on the surfaces of titanium alloy and(0.382?0.075)?105 CFU on Co-Cr-Mo alloy.When each group compared with UHMWPE and domestic PMMA respectively,all P value was

OBJECTIVE To determine the feasibility of clindamycin-loaded calcium phosphate cement(CLCPC) as a local antibiotic delivery system.METHODS The initial setting time(tI) and the final setting time(tF) were measured for 0%,2% and 5% CLCPC according to ASTM C266-89 method.Clindamycin concentrations eluting from the samples of 2% and 5% CLCPC in PBS were analyzed by HPLC at different times.The bacteriostasis tests were done by plate diffusion method for 2% and 5% CLCPC and 2% Palacos R-40 bone cement(PMMP) samples,and the diameters of the bacteriostasis ring and bacteriostasis duration were observed.The setting product and crystal size of 0%,2% and 5% CLCPC were analyzed and observed by X-ray diffraction(XRD) and scanning electron microscopy(SEM).RESULTS The setting time could be shortened by adding clindamycin(tI,tF) of 2% and 5% CLCPC.Clindamycin was with burst-release from CLCPC within the intial 6-hour period and the release rate slowed down on 4th day.Clindamycin could still release until to the 42th day.The ring of 2% Palacos R-40 bone cement(PMMP) bacteriostasis was smaller than that of 2% CLCPC,and the ring of 2% CLCPC bacteriostasis was smaller than that of 5%CLCPC.The bacteriostasis still existed to the 42th day of test for 2% and 5%CLCPC and 2% Clindamycin-loaded Palacos R-40 bone cement(CLPMMP).From the 30th day,many bacterial colonies were seen in the culture media laying 2%CLPMMP sample.On the contrary,bacterial colony was not found in the media putting 2% and 5%CLCPC.XRD and SEM showed that clindamycin did′t have an influence on setting product,crystal size and structure of CPC.CONCLUSIONS Clindamycin-loaded calcium phosphate cement can be used as a local antibiotic delivery system.