BACKGROUND: Revascularization is necessary for tissue-engineered bone implantation by osteogenesis to effectively repair bone defect.OBJECTIVE: To evaluate marrow mesenchymal stem cells (MSCs) modified by bone morphogenetic protein 2 (BMP-2) in combination with polylactic acid/polycaprolactone (PLA/PCL) to repair rabbit radial bone defect during the vascularization, and to investigate the promotive effects of BMP-2 gene on the vascularization of bone graft.DESIGN: Randomized controlled animal study.SETTING: This study was performed in the Central Laboratory of China Medical University from January to December 2005.MATERIALS: PLA/PCL with 150-250 μm pore diameter and 90% interval porosity was provided by Changchun Applied Chemistry Institute, Chinese Academy of Science. Sixty 3-month-old New Zealand rabbits were selected in this study.METHODS: Sixty rabbits were randomly divided into four groups with 15 rabbits in each group. Subsequently, middle segments of bilateral radial bone were obtained to establish 1.5-cm bone defect models that were implanted with processed artificial bones. Adenovirus carrying BMP-2 (AD-BMP-2) group: Artificial bones were processed with transfected BMP-2 cells plus PLA/PCL; Control group: Artificial bones were processed with adenovirus carrying β-gal gene (Ad-Lacz) plus PLA/PCL; Non-transfection group: Artificial bones were processed with non-transfected cells plus PLA/PCL; PLA/PCL group: PLA/PCL alone for transplantation.MAIN OUTCOME MEASURES: Four, eight, and twelve weeks after surgery, X-ray was used to observe new bone formation; stereoscopic microscope to observe distribution of microvessels; haematoxylin-eosin staining to detect the relationship between microvessels and bone formation; transmission electron microscope to investigate the correlation between osteoblasts and vascular endothelial cells, detect vascular endothelial growth factor expression, and calculate the number of microvessels.RESULTS: Four postoperative weeks in the AD-BMP-2 group, numerous microvessels were observed; stent pore was full of cartilage calluses; active osteoblasts grew around microvessels; vascular endothelial growth factor expression and numbers of microvessels were higher and more than those in other groups. Eight postoperative weeks, osteoblasts gradually increased in the bone graft; microvessels circuitously expanded and connected each other; cartilage callus changed into trabecular bone. Twelve postoperative weeks, cortical bones were successive; medullary cavity recanalized; microvessels longitudinally arranged in order. Ability of bone formation in the control group and non-transfection group was weak, and vascular regeneration was slow; 12 postoperative weeks, bone defect was primarily repaired; microvessels were distributed along the pores of newborn bone trabecula. Newborn vessels were hard found in the PLA/PCL group at each time point. Twelve postoperative weeks, bone extremities sclerotized, and defect regions were fully filled by fiber tissues.CONCLUSION: Transfected BMP-2 gone by up-regulating vascular endothelial growth factor expression can indirectly induce vascularization of bone graft, promote survival of seed cells, and accelerate bone formation.

BACKGROUND: Tissue engineered bone constructed in vitro is a compound of cell and material; additionally, revascularization plays a key role in effectively repairing bone defect after transplantation of tissue engineered bone.OBJECTIVE: To evaluate the influence of antigen-extracted bovine cancellous bone stent combining with bone morphogenetic protein 2 (BMP-2) on vascularized reaction during reparative process of bone defect.DESIGN, TIME AND SETTING: Randomized grouping design and controlled animal study, which was performed at the Central Laboratory, China-Japan Friendship Hospital Affiliated to Jilin University.MATERIALS: Sixty rabbits of clean grade were selected in this study. Superior cancellous bone of bovine humerus was used to establish bovine cancellous bone stent. Recombinant adenovirus carrying human BMP-2 (Ad-BMP-2) and β -galactosidae gene (Ad-Lacz), and recombinant human BMP-2 (rhBMP-2) were graciously presented by Dr. Oliver and Pro. Gao, Department of Pathology of Jilin University.METHODS: Marrow mesenchymal stem cells were extracted from 60 rabbits, and then they were transfected with BMP-2 adenovirus vector to repair 1.5-cm defects of radial bone of both upper extremities by combining with bovine cancellous bone transplantation. Rabbits were randomly divided into 5 group with 12 in each group: Ad-BMP-2 transfected cells+bovine cancellous bone group, non-transfected cells+rh-BMP-2+bovine cancellous bone group, Ad-Lacz transfected cells +bovine cancellous bone group, non-transfected cells + bovine cancellous bone group, and bovine cancellous bone group. Transplanted bone was fixed by tightly suturing tunica muscularis and anadesma.MAIN OUTCOME MEASURES: New bone formation was observed by X ray at 4, 8, and 12 weeks after surgery; microvascular ink perfusion was used to observe vascular distribution; transmission electron microscope was used to observe osteoblasts and vascularization; hematoxylin-eosin (HE) staining, alcian blue staining, and VonKossa staining were used to observe correlation between microvessels and bone formation; immunohistochemical staining of vascular endothelial growth factor was used to detect gray value; immunohistochemical staining of CD34 was used to specifically label vascular endothelial cells for microvascular amount.RESULTS: Sixty rabbits were included into the final analysis. X-ray and immunohistochemical examinations demonstrated that bone formation and vascularization in the Ad-BMP-2 transfected cells+bovine cancellous bone group and non-transfected cells+rh-BMP-2+bovine cancellous bone group were superior to those in other three groups. After four weeks, microvascular ink perfusion indicated that a branch of small vessels was formed in pore of trabecular bone. Vascular density was higher in the peripheral domains but lower in the central regions. Transmission electron microscope suggested that a lot of osteoblasts and new vascular buds with active function were observed four weeks after surgery. Mature lamellar bone was formed 12 weeks after surgery, and structure of new vessels was complete. Detection of vascular endothelial growth factor expression and microvascular amount indicated that content of vascular endothelial growth factor in the Ad-BMP-2 transfected cells+bovine cancellous bone group was significantly higher than that in other four groups (P<0.01), and microvascular amount was also significantly higher than that in other four groups (P<0.01).CONCLUSION: BMP-2 gene can indirectly induce vascularization of transplanted bone through up-regulating vascular endothelial growth factor expression, which is superior to rh-BMP-2.

BACKGROUND:The incorporating of zeolites or porous fillers into polymer membranes can improve the pervaporation separation properties of membranes.But the effect mechanism of zeolites or porous fillers on membrane properties needs to be further studied.OBJECTIVE:The study was designed to prepare ZSM-5 zeolite incorporated chitosan membrane by solution blending method to investigate the effecIs of zeolite on membrane pervaporation properties.DESIGN:A controlled observation.SETTING:Dalian Institute of Chemical Physics,Chinese Academy of Sciences.MATERIALS:This study was performed at the Laboratory of New Type Membrane Separation Technology,Dalian Institute of Chemical Physics,Chinese Academy of Sciences in May 2006.Chitosan,with deacetylation degree of 75%-85%,was obtained from Sigma-Aldrich Chemical Company,USA;Dimethyl carbonate(DMC),with purity of 99%,was purchased from Fluka Chemical Company,USA:ZSM-5 zeolite was kindly supplied by Dalian Institute of Chemical Physics,Chinese Academy of Sciences,China;Methanol,with purity＞99.5%,were purchased from Shenyang Lianbang Chemical Company,China.METHODS:ZSM-5 zeolite incorporated chitosan membranes were prepared by solution blending method.properties.RESULTS:Scanning electron mieroscope demonstrated that zeolite was uniformly distributed in the membrane matrix and the membranes were free from possible defects.The separation selectivity of dimethyl carbonate(DMC)/methanol mixtures was dominated by solubility selectivity rathcr than diffusivity selectivity.Swelling degree increased and the permeation flux of the membranes increased significantly with the zeolite content increasing.From the temperature-dependent permcation values,the Arrhenius activation parameten were estimated.CONCLUSION:The pervaporation results indicated that the membranes incorporated with the ZSM-5 zeolite exhibited better separation properties for DMC/methanol mixtures comparing with homogeneous chitosan membranes.

Objective To investigate the effect of bFGF on the grafting ability (grafing efficiency)of CD34 + cord blood cells seeded in 5 FU injured human bone marrow stromal cells.Methods Using combined methods of bone marrow stromal cell culture system,light microscopy and electron microscopy,flow cytometry,we investigated the toxic effects of 5 FU on human bone marrow stromal cells (hBMSC) and the benefit of bFGF on dynamics of hBMSC as well as the ability of the cord blood stem cell to bind hBMSC.Results After 3 hours incubation with 5 FU,significant changes in hBMSC cytosolic and nuclear morphology were observed.Nuclear splitting and abnormal shapes of the pseudopodia were found under the electron microscope.Flow cytometry revealed that apoptosis appeared before G 0/G 1,and that cells in G 0/G 1and G 2/M phases were remarkably lower,while the cells in S phase were remarkably higher than the controls.Two hours after adding the cord blood hematopoietic progenitor cells,the group treated with bFGF(injury with intervention of bFGF group,IBG)showed higher proprtion of the adherence layer compared to the group without adding bFGF(injury group,IG)(29% versus 12%) Conclusion 5 FU causes DNA damage in hBMSC.It also impairs the binding ability of cord blood stem cell to hBMSC.bFGF is able to repair hBMSC damage and improve the binding of CD34 + cord blood hematopoietic progenitor cells onto hBMSC.

Objective To explore the expression of mismatch repair gene (MMR) (MLH1,MSH2,PMS2 and MSH6) in endometrioid adenocarcinoma,and to analyze its clinical pathological significance.Methods Inmunohistochemical EnVision method was used to detect the expressions of DNA mismatch repair proteins (MLH1,MSH2,PMS2 and MSH6) in 101 cases of endometrial adenocarcinoma.The phenotypes of tumor microsatellite instability (MSI) and microsatellite stable were determined,and its relationship with onset age,differentiation,depth of myometrial invasion,lymphatic metastasis and prognosis of tumor were analyzed.Results There were 67 cases of hysterectomy and 34 cases of endometrial biopsy in 101 cases of endometrial adenocarcinoma.52 cases of surgical specimens and 14 cases of biopsy specimens were included in 66 cases that had complete follow-up data.Rates of negative expression were as follows:MLH1 31.6 ％ (32/101),MSH228.7 ％ (29/101),PMS2 16.8 ％ (17/101),MSH6 8.9 ％ (9/101).MSI phenotype 53.5 ％ (54/101),MSS phenotype 46.5 ％ (47/101).Univariate analysis showed that prognoses of MSI-L&MSS group (34 cases) were better than those of MSI-H group (18 cases) in hysterectomy organization (P =0.041).The differences between depth of myometrial invasion,degree of differentiation,age and prognosis of endometrial adenocarcinoma were statistically significant (P ＜0.05).Cox multivariate analysis showed that differences between depth of myometrial invasion,age and prognosis of endometrial adenocarcinoma were statistically significant (P =0.034,P =0.009),while there was no significant difference between MSI-L&MSS group and MSI-H group in prognosis (P ＞ 0.05).Conclusions MSI is one of the molecular events that occur in the endometrioid adenocarcinoma.Age and depth of tumor invasion are independent prognostic factors of endometrioid adenocarcinoma.In view of inconsistency between univariate analysis and multivariate analysis,whether MSI can be used as deterministic accordance for endometrioid adenocarcinoma prognostic evaluation requires further verification.

OBJECTIVE To explore the method for sterilizing hands which is effective,convenient and without stimulation for medical personnel.METHODS A total of 150 nurses on duty were divided into three groups randomly.In the test groups 1and 2,towels with 0.2% peracetic acid and BANENG disinfectant fluid were used separately,but in the control group only soap with flowing water was used for hands sterilization.RESULTS The rate of bacterial removal was 98.82%,72.63% and 68.64%,respectively,among the 3 groups.The efficacy of BANENG disinfectant fluid was similar to 0.2% peracetic acid.Both of them showed significant differences compared with control group.CONCLUSIONS Both 0.2% Peracetic acid and BANENG disinfectant fluid have better disinfection efficacy,and BANENG disinfectant fluid is more easier in use.

Objective To explore clinical therapeutic effect of cattle encephalon glycoside and ignotin (EGI)on adjunc-tive therapy of severe hand-foot-and-mouth disease (HFMD).Methods 94 children with severe HFMD in a hospital were selected and divided into two groups according to the parity of patients’admission numbers,46 children in control group were given conventional therapy;48 children in trial group were given EGI adjunctive therapy based on conventional thera-py,therapeutic effect of two groups was compared.Results The total therapeutic effective rate in trial group was signifi-cantly higher than that of control group (95.83% vs 76.09%,χ2 =16.81,P <0.001 ).Neurological function score was not significantly different between two groups before therapy(P >0.05);after therapy,neurological function score in trial group was significantly lower than control group ([4.07±0.13]vs [5.59±0.18],χ2 =18.19,P <0.05).The recovery time of temperature,heart rate,blood pressure,and breathing,as well as length of stay in hospital in trial group were all significantly shorter than control group(P <0.05);incidence of complications in trial group was lower than control group (4.17%[n=2]vs 21.74%[n=10],χ2 =10.08,P <0.001 ).The proportion of turning into critically ill cases in trial group was significantly lower than control group (2.08% [n = 1 ]vs 13.04% [n = 6 ],χ2 = 8.94,P < 0.001 ). Conclusion EGI has remarkable efficacy in adjunctive therapy of severe HFMD,which can effectively improve clinical symptoms,and is worthy to be popularized and applied.

The technique of chimeric antigen receptor-engineered T cells (CAR-T) is combined single chain antibody and T-cell activation modification and has specific and durable killing effect on tumors.It had achieved stunning results on clinical trials of B lymphocytic leukemias,B lymphomas and solid tumors.Meanwhile it also had potential risk on the off target effects,cytokine release syndrome,graft versus host disease and so on.This paper briefly reviews the principal of CAR-T cellular mechanism,advantages and clinical applications.

OBJECTIVE To investigate the changes in and clinical significance of C-reactive protein(CRP) and endotoxin in patients who are firstly examined in fever clinic.METHODS To review and study the results of CRP and endotoxin from 81 patients(including 48 patients who were diagnosed as bacterial pneumonia and 33 patients who were diagnosed as a cold) in fever clinic of our hospital in 2005.RESULTS In 81 bacterial infection patients, the analysis indicated the levels of endotoxin between the pneumonia patients and the patients with a cold had significant difference.And the levels in the first group were significantly higher than in the latter one.The levels of CRP had no difference in two groups.CONCLUSIONS The result indicates plasma endotoxin detection helps to determine the early diagnosis of serious infection.It is also of significance to select antibacterial rationally and handle syndrome in time.In fever clinic,the levels of endotoxin may be a method to help diagnosis and treatment.CRP could be help to estimate the bacterial infection or virus infection,but it could not adjudge the seriousness in bacterial infection.

Objective To analyze the clinical value of serum and cerebrospinal fluid(CSF) procalcitonin in the diagnosis of adult central nervous system(CNS) infection to provide reference for its clinical diagnosis and treatment .Methods Sixty adult patients with CNS diseases admitted to our hospital from February to December 2015 were selected as the research subjects .All patients were suspected of CNS infection at initial visiting due to suspected CNS symptoms and signs ,and the CSF pathogenic examination and separation were performed to determine the types of infected pathogenic microorganisms .Meanwhile serum and CSF procalcito-nin was detected .The detection results of CSF pathogen and procalcitonin were summarized .The detection results of CSF pathogen served as the gold standard ,serum and CSF procalcitonin> 0 .5 ng/mL as infection positive .The diagnostic test fourfold table was adopted .Then the procalcitonin detection results and CSF pathogenic microorganism detection data were inputted .The consistency of serum and CSF procalcitonin for judging CNS infection was analyzed and calculated .The sensitivity ,specificity and accuracy of serum and CSF procalcitonin detection method for detecting pathogenic microorganisms were analyzed .Results Among 60 cases ,31 cases were bacterial infection ,22 cases were viral infection and no pathogenic bacterium was detected out in 7 cases ,the positive rate was 88 .34% .In the serum procalcitonin detection :49 cases were positive and 11 cases were negative ;in CFS procalcitonin detec-tion:37 cases were positive and 23 cases were negative .The Kappa coefficients of serum procalcitonin and CFS pathogenic detection results was 0 .769 ,indicating that serum procalcitonin and CFS pathogenic detection results had higher consistency ;the Kappa coef-ficients of CSF procalcitonin and CFS pathogenic detection results was 0 .315 ,indicating that CFS procalcitonin and CFS pathogenic detection results had poor consistency .The sensitivity ,accuracy and specificity of serum procalcitonin for diagnosing CNS infection were 96 .08% ,93 .33% and 77 .78% respectively ,which of CSF procalcitonin were 73 .33% ,45 .00% and 38 .46% respectively ;se-rum procalcitonin detection had a higher clinical value in the diagnosis of adult CNS infection ,while CSF procalcitonin detection had little value .Conclusion The serum procalcitonin detection method has higher sensitivity ,accuracy and specificity for the diagnosis of adult CNS infection ,possesses a better guidance role in selecting the treatment scheme and evaluating the curative effect ,and has higher clinical value in adult patients with CNS bacterial infection .