[Objective]To treat the femoral neck fracture using cannulated screw and BMP release system in animals,and to evaluate the possibility of treating femoral neck fracture and provide experimental basis for clinical application.[Method]Eighteen mongeral were used in this study.The model of bilateral femoral neck dislocation fracture was established.The control side was fixed using cannulated screw,and the experiment side was fixed with cannulated screw and injectable BMP release system.At 4,8,12 weeks,6 animals were sacrificed at one time point respectively.Results were obtained through histology,radiography,scintimetry and gross observation.[Result]The fracture line was vague at four weeks,and at eight weeks the fracture line almost disappeared.It was healed completely at twelve weeks.Radiological study showed that the healing of the fracture in experimental side was better than that of the control side.There was the same result of observation in histology.[Conclusion]The cannulated screw combined with BMP used in the experiment is effective and feasible.It may not only provide strong internal fixation but also infuse growth factor into site of fracture.It would accelerate the reconstructing of the vascular supply to the femoral head after the fracture and promote the restoration of bone.

Adenoma ; Adult ; Ear Neoplasms ; Ear, Middle ; pathology ; Female ; Humans

Objective To investigate the effects of iodine excess on spleen cell viability,lactate dehydrogenase (LDH) leakage,mitochondrial superoxide production and peroxiredoxin (Prx)3 expression in methallothionein Ⅰ / Ⅱ knockout (MT-Ⅰ / Ⅱ KO)mice.Methods Spleen cell suspensions were prepared from six to eight-week old and healthy male MT-Ⅰ / Ⅱ KO mice and wild type (WT) mice; the cell number was adjusted to 5 × 107/L and the cells were plated in 96-well plates (100 μl each well); the cells were exposed to various concentrations of KI (0,10-4,10-3,10-2 mol/L) and 10-3 mol/L H2O2,respectively,for two hours,and control group did not give KI nor H2O2.Cell viability was assayed by methyl thiazolyl tetrazolium (MTT) colorimetric method.Cell damage was detected by chemical colorimetric method.Mitochondrial superoxide production in the spleen cells was measured by flow cytometry.Western blotting technology was used to investigate the expression of Prx3.Results In both MT-Ⅰ/Ⅱ KO and WT mice,the differences of cell viability,LDH leakage,mitochondrial superoxideproduction and the expression of Prx3 of spleen cells among the treatment groups were statistically significant (F =357.92,71.03,130.36,10.36,179.58,26.92,187.43,and 7.16,all P ＜ 0.05).Compared to the control group [(100.00 ± 2.00)％,(100.00 ± 1.63)％,(3 202.22 ± 85.63),(3 161.51 ± 144.49)U/L,43.82 ± 1.56,38.60 ± 2.81,0.61 ± 0.09,0.50 ± 0.08],cell viability of 10-4,10-3,10-2 mol/L KI treatment and 10-3 mol/L H2O2 groups [(80.77 ± 1.86)％,(89.89 ± 2.90)％,(76.08 ± 1.92)％,(87.66 ± 1.74),(73.26 ± 1.86)％,(84.30 ± 2.23)％,(66.22 ± 1.71)％,(70.80 ± 1.49)％] was decreased (all P ＜ 0.05); LDH leakage [(3 880.00 ± 190.62),(3 431.17 ± 170.45),(4 178.33 ± 170.43),(3 598.63 ± 189.09),(4 388.61 ± 123.79),(3 863.72 ± 195.64),(4 615.28 ± 196.17),(4 148.12 ± 195.81)U/L] was increased significantly (all P＜ 0.05); and mitochondrial superoxide production in the spleen cells (53.83 ± 3.22,47.03 ± 1.60,58.92 ± 4.00,50.48 ± 2.59,72.72 ± 2.14,68.53 ± 2.97,80.76 ± 4.11,75.26 ± 3.41) was increased significantly (all P ＜ 0.05); Prx3 expressions in 10-3、10-2 mol/LKI and 10-3 mol/L H2O2 treatment groups (0.82 ± 0.12,0.65 ± 0.12,0.96 ± 0.15,0.73 ± 0.16,1.04 ± 0.13,0.85 ± 0.16) significantly increased (all P ＜ 0.05),the differences of Prx3 expressions between 104 mol/L KI groups (0.73 ± 0.15,0.55 ± 0.09),and control groups were not statistically significant (all P ＞ 0.05).In 104,10-3,10-2 mol/L KI and 10-3 mol/L H2O2 treatment groups,cell viability of MT-Ⅰ/Ⅱ KO mice spleen was lower than that of WT mice (t =6.47,10.93,9.30 and 4.96,all P ＜ 0.05); LDH leakage was higher than that of WT mice (t =4.30,5.58,5.56 and 4.13,all P ＜ 0.05); mitochondria superoxide production was higher than that of WT mice (t =4.64,4.33,2.80 and 2.52,all P ＜ 0.05); Prx3 expression was higher than that of WT mice (t =2.54,2.37,2.59 and 2.27,all P ＜ 0.05).Conclusions KI may decline the cell viability,increase the leakage of LDH and increase the production of mitochondrial superoxide production and Prx 3 expression,which are much more significant in MT-Ⅰ /Ⅱ KO mice,suggesting that MT Ⅰ /Ⅱ has some antioxidative effect in high concentration of iodide induced oxidative stress in the spleen.

BACKGROUND: Traditional methods of repairing bone defect such as autograft and allograft have some disadvantages that are hard to deal with, gene treatment may be a new approach. OBJECTIVE: To investigate the biological properties of cultured human osteoblasts transfected with vascular endothelial growth factor (VEGF) gene. DESIGN, TIME AND SETTING: A controlled experiment based on cytology was carried out in the Scientific Experiment Centre of Liaoning Medical College from May 2005 to May 2006. MATERIALS: Human lilac born block was harvested from a cervical spondylosis patient who required lilac bone graft with his informed consent of this patient. Plasmid pCDI/VEGF_(121) was given as gift from Professor Ma, Peking Unviersity Human Disease Genomics Research Center. Competent Escherichia coU was given as gift from Professor Liu, Liaoning Medical College. METHODS: Human osteoblasts were isolated and cultured in vitro. There were a VEGF transfection group and a control group in the experiement. Using cation liposome, the pCDINEGF_(121) eukaryotic expression plasmis was induced into human osteoblasts.MAIN OUTCOME MEASURES: At 1, 3, 5, 7 days following passage culture, the expression of VEGF in human osteoblasts was detected. Its effects on the call proliferation, the secretion of osteocalcin and alkaline phosphatase were investigated.RESULTS: After the plasmid pCDI-VEGF_(121) was transferred into human osteoblasts 3 and 7 days, VEGF mRNA expression was detectable by RT-CPR method. The call number of transfection group was larger than that of control group (P < 0.05 or P < 0.01).When the cells were cultured for 3 days, the positive rate of alkaline phosphatase in the transfection group was increased compared with control group (P < 0.01 ); the secretion of osteocalcin in the transfection group was higher than that of control group (P < 0.05 or P < 0.01 ).CONCLUSION: VEGF gene transfection can improve the proliferation and biological function of human osteoblasts cultured in vitro.

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 explore the expressions of survivin and Ki-67 in breast cancer tissues after neo-adjuvant chemotherapy and their clinical significance.Methods The expressions of survivin and Ki-67 in 60 cases of breast cancer with neo-adjuvant chemotherapy(NACT) with CTF regimen and 60 cases without NACT were detected by SABC immunohistochemical technique.ResultsThe positive rates of survivin(36.7%)and Ki-67(38.3%) in neo-adjuvant chemotherapy group were significantly lower than those in control group(71.7%,61.7%)(P0.05).ConclusionsThe therapeutic effect of neo-adjuvant chemotherapy with CTF regimen can increase the remission rate of breast cancer.Neo-adjuvant chemotherapy could inhibit the expression of survivin protein,and thus further inhibit proliferation of tumor cell.

Objective To detect the mRNA and protein expression of downstream quinine nicotinamide adenine dinucleotide (NADH) dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) induced by blood nuclearrelated factor E2 (Nrf2) in the peripheral blood of those exposed to arsenic in the endemic area of coal arsenic poisoning in Guizhou Province,and to discuss its role in the process of occurrence and development of liver injury due to coal arsenic poisoning.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey sites,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group based on physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.Peripheral blood samples were collected from all subjects.And mRNA expression of NQO1 and HO-1 were detected by RT-qPCR.Content of NQQ1 and HO-1 were detected by enzyme linked immunosorbent assay (ELISA).Results (①)Results of mRNA expression of NQ01 and HO-1:the relative expression level of mRNA of NQO1 and HO-1 in peripheral blood went up gradually as the degree of liver injury of population exposed to arsenic increased (F =5.548,10.961,all P ＜ 0.05);thereinto,the relative expression level of mRNA of NQO1 of mild hepatosis group (median:0.918) was higher than that of the control group (0.576,P ＜ 0.05),the relative expression level of mRNA of NQO1 of moderately severe hepatosis group (1.243) was higher than those of control group,non-patient group (0.653),non-apparent hepatosis group (0.636) and mild hepatosis group (all P ＜ 0.05);the relative expression level of mRNA of HO-1 of non-patient group (1.059),non-apparent hepatosis group (1.225),mild hepatosis group (1.553) and moderately severe hepatosis group (1.604) were higher than that of control group (0.767,all P ＜ 0.05);the relative expression level of mRNA of HO-1 of mild hepatosis group was higher than that of non-patient group (P ＜ 0.05);the relative expression level of mRNA of HO-1 of moderately severe hepatosis group was higher than those of non-patient group and non-apparent hepatosis group (all P ＜ 0.05).②)Results of protein expression of NQO-1 and HO-1:the level of protein expression of NQO1 and HO-1 in serum went up gradually as the degree of liver injury of population exposed to arsenic increased (F =19.685,17.725,all P ＜ 0.05).Thereinto,the protein expression of NQO1 and HO-1 of non-apparent hepatosis group,mild hepatosis group and moderately severe hepatosis group [NQO1:(6.272 ± 0.744),(6.336 ± 0.628),(6.714 ± 0.540) U/L;HO-1:(45.150 ± 4.813),(45.283 ± 5.049),(48.610 ± 5.365) U/L] were higher than those of control group and non-patient group [NQO1:(5.550 ± 0.730),(5.750 ± 0.427) U/L;HO-1:(39.856 ± 5.249),(42.375 ± 3.014) U/L,all P ＜ 0.05],the protein expression of NQO1 and HO-1 of moderately severe hepatosis group were higher than those of non-apparent hepatosis group and mild hepatosis group (all P ＜ 0.05).Conclusion The expression of mRNA and protein of NQO1 and HO-1 is closely related to the occurrence and development of liver injury due to arsenic exposure in coal arsenic poisoning areas.

Objective To define the indications and evaluate the results of various surgical treatment options in patients with nonparasitic hepatic cysts. Methods The clinical data of 284 patients with nonparasitic hepatic cysts treated in our hospital from January 1995 to December 2005 were retrospectively analyzed. Results Percutaneous aspiration and sclerotherapy were performed in 161 cases and complications occurred in 9 (5.59%), recurrence in 53 (32.92%) and the mortality was 0%.Open surgery was conducted in 71 cases and the complications occurred in 16 (22.54%), recurrence in 8 cases (11.27%) and the mortality was 2.82%. Laparoscopic surgery was employed in 52 cases and complications occurred in 7 ( 13.46%), recurrence in 6 (11.54%) and the mortality was 1.92%.Conclusion There is currently no general agreement in the literature concerning when nonparasitic hepatic cysts should be treated. Laparoscopic surgery was more favorable than other therapeutic options.However, we should choose individually suitable methods according to different clinical symptom of patients.

Objective To study Chinese medicine and Western medicine preventing the anaphylaxis of ionic-type radiography agent,to solve the anaphylaxis in brain enhancment CT.Methods Eight hundred and forty cases have been diagnosed in brain enhancement CT at our hospital,from Fed.1995 to May 2000.Results Chinese medicine,ginger,pinellia and hormone medicine were all useful in preventing the anaphylaxis of ionic-type radiography agent in brain enhancement CT.Conclusion Chinese medicine:ginger and pinellia could prevent the anaphylaxis of ionic-type radiography agent.

Aim Vascular endothelial cell growth inhibitor(VEGI) is a recently discovered novel member of the TNF superfamily,which is expressed predominantly in endothelial cells.As an endothelial cell-specific negative regulator of angiogenesis,the relationship between structure and function of VEGI is not understood at present.Methods In order to explore the functional key amino acids of VEGI,four mutants of VEGI(E45→R,G47→A,Y111→F,Y111→T) were construced by site-directed mutagenesis,and recombinant proteins were generated from E.coli.Four mutant proteins behaved similar to the wild type VEGI in various physico-chemical assays.The proliferation of HUVEC and chick choriallantic membrane assay were performed to study the activity of four mutants.Results The mutant E45→R significantly decreased the biological activity,and the mutant G47→A caused a slight drop on activity,but the mutants Y111→F,Y111→T almost completely abolished biological activity.Conclusion It suggests that Y111 is an important residue in biological activity,which may play a direct role in receptor recognition.Moreover,the tyrosine ring and hydroxy group of the amino acid are important determinant of biological activity.Additionally,E45 also plays an important role in biological activity of VEGI.