OBJECTIVE To understand the feature of nosocomial infection in cerebral palsy and effective contermeasure s. METHODS Prospective and retrospective studies on 1272 hospitalized patients in recent two years were conducted. RESULTS Incidence rates of hospital infection that were calculated by the number of the patients and the cases were 9.28% and 9.75%.Two major infection sites were the respiratory and the gastrointestinal tracts.The primary disease was respiratory infection(47.46%) with increase in younger group and seasonaly change. CONCLUSIONS Cerebral palsy children are susceptible to the infection.Doctors should pay attention to the risk factors to prevent the occurrence of the infection.

Aim To investigate the effect of paeoniflorin(PF)on TLR2/4 pathway in AGEs-induced RAW264.7 macrophages.Methods RAW264.7 macrophages were incubated at different time points in AGEs stimulation,as well as different concentrations of PF,to optimize experimental conditions.RAW264.7 macrophages were randomly divided into five groups: control group(DMEM),bull serum albumin(BSA)group(200 mg·L-1 BSA),AGEs group(200 mg·L-1 AGEs),paeoniflorin group(200 mg·L-1 AGEs+10-5 mol·L-1 PF)and TLR2/4 inhibitor group(200 mg·L-1 AGEs+30 mg·L-1 OxPAPC).The expression of Toll-like receptor 2(TLR2),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),p-IRAK1,TIR-domain containing adaptor protein-inducing IFN-β(TRIF),interferon regulatory factor 3(IRF3),p-IRF3,NF-κB p-p65,NF-κB p65,inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),interleukin-l β(IL-1β)and monocyte chemotactic protein-1(MCP-1)were measured by Western blot.Real-time PCR was used to detect the expression of TLR2 and TLR4 mRNA,while TNF-α,IL-1β and MCP-1 levels in cell supernatant were measured by ELISA.Results Compared with control group,AGEs significantly increased the expression of TLR2,TLR4,MyD88,p-IRAK1,TRIF,IRF3,p-IRF3,NF-κB p-p65,NF-κB p65,iNOS,TNF-α,IL-1β and MCP-1 proteins(P<0.01),as well as TLR2 and TLR4 mRNA(P<0.01).TNF-α,IL-1β and MCP-1 contents were also elevated in cell supernatant(P<0.01).The effects induced by AGEs were decreased significantly in PF and TLR2/4 inhibitor group(P<0.01).Conclusion PF plays an anti-inflammatory effect via inhibiting TLR2/4 pathway on macrophages,which may provide a new theoretical basis for the treatment of diabetic nephropathy.

A simple and reliable high performance liquid chromatography (HPLC) method has been developed for the simultaneous quantification of five major bioactive components in ‘Shu-Jin-Zhi-Tong' capsules (SJZTC), for the purposes of quality control of this commonly prescribed traditional Chinese medicine. Under the optimum conditions, excellent separation was achieved, and the assay was fully validated in terms of linearity, precision, repeatability, stability and accuracy. The validated method was applied successfully to the determination of the five compounds in SJZTC samples from different production batches. The HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of SJZTC.

Objective To measure and analyze Varian Clinac 21EX accelerator induced radioactivity,and to provide specific recommendations and ways of radiation protection for radiotherapy technicians.Methods To simulate the working environment of radiotherapy technician,and to detect induced radioactivity variation of Varian Clinac 21EX accelerator that induced by 15 MV X-rays under different conditions of beam field area,dose,time,distance and by high energy electron beam (12,16,20 MeV)at the different time.Results The induced radioactivity level was not influenced by different beam field area,and was increased with increasing dose (r =0.930,P ＜ 0.05),decreased with time increase (r =-0.84,P ＜ 0.05),decreased with distance increase(r =-0.975,P ＜ 0.05).The induced radioactivity attenuation levels of the different doses at the same time are different,and have the common characteristic that the induced radioactivity attenuation rate is faster in the initial times.The induced radioactivity levels of high energy electron beams were significantly lower than those of high-energy X-rays.Conclusions During radiotherapy positioning,it is necessary to take measures to protect against the induced radioactivity when high-energy rays with energy greater than 10 MeV will be used.The radiotherapy technician should take the different time and energy interval steps for the different ray type and energy and dose in order to meet the radiation protection principle of optimization.

Objective To investigate the utility of combined test of neutrophil CD64 and C-reactive protein(CRP)for early diag-nosis of burn sepsis.Methods A prospective study was conducted in 76 hospitalized burn patients,including three groups:burn sepsis(n=27),local infection(n=31)and controls without bacterial infection (n=18)based on their diagnosis.Blood samples were collected within 24-48 h after hospital admission for analysis of neutrophil CD64,CRP,white blood cell count (WBC), neutrophils percentage (N)by flow cytometry,and bacterial culture.These parameters were re-evaluated for the patients in sepsis group 7 and 14 days after antibiotic therapy.Results Neutrophil CD64,CRP,WBC and N were significantly higher in sepsis patients than the corresponding values in the patients with local infection or control patients (P < 0.01 ).Neutrophil CD64 and CRP in the patients with local infection were significantly higher than those in the control patients without bacterial infection (P < 0.01).Neutrophil CD64 was positively correlated to CRP.Neutrophil CD64 and CRP decreased 7 days after an-tibiotic therapy,but the difference was not statistically significant compared to the pretreatment levels.Neutrophil CD64 and CRP decreased significantly 14 days after antibiotic therapy compared to the pretreatment levels (P < 0.01),but still higher than the levels in the control group.The receiver operating characteristic (ROC)curve of CD64 + CRP combined test,CD64, CRP,WBC,N for detecting bacterial infection showed an area under curve (AUC)of 0.952,0.923,0.904,0.756 and 0.728,respectively.Conclusions Neutrophil CD64 is useful in early diagnosis of burn sepsis.The expression of neutrophil CD64 does not have significant difference between gram-positive and gram-negative bacterial infections.Combined test of neutrophil CD64 and CRP can improve the efficiency for diagnosis of burn sepsis.

Purpose To investigate the clinical characteristics and Oxford classification of IgA nephropathy patients with hyperurice-mia. Methods 151 IgA nephropathy patients confirmed by renal biopsy in 2013 were analyzed retrospectively. The patients were di-vided into the two groups:IgA nephropathy patients with or without hyperuricemia. Morphological changes were evaluated with Oxford classification scoring system and Lee’s grades. A comparative analysis of clinical manifestations and pathological injuries was performed between the two groups. Results Incidence of hyperuricemia in IgA nephropathy patients was 48. 3% and was more common in young men. Hypertension was associated with hyperuricemia. Oxford classification of IgA nephropathy patients with hyperuricemia was pre-dominant M1E0S1T0 and Lee’s grades presented with grade Ⅲ. The outstanding histopathologic features with higher plasma uric acid levels indicated higher tubulointerstitial chronicity, higher glomerular sclerosis ratio, accompanied by a decline in glomerular filtration rate. There was no significant difference of vascular lesions. Conclusions The prevalence of hyperuricemia in IgA nephropathy pa-tients is high. Oxford classification shows IgA nephropathy with hyperuricemia are associated with more severe tubulointerstitial lesions and lower GFR.

Objective To explore the relationship of X-ray exposure parameters and false-node rate during image-guidance treatment with CyberKnife spine tracking.Methods Using spine tracking planning on a chest phantom,several combinations of X-ray exposure parameters were used to locate.The false-node ratio and the surface absorbed dose were investigated and the radiation dose was optimized.Results The false-node ratio and surface absorbed radiation dose decreased when the X-ray exposure parameters increased until they saturated.In the range of ≤5.0％ false-node rate,the surface absorbed radiation dose was 0.11,0.26 mGy,and 0.31-0.46 mGy,when the false-node rate was 2.77％,1.07％and 1.0％,respectively.Conclusions In image-guided treatment of CyberKnife,the radiation dose would be optimized,and the patient's radiation dose would be reduced greatly,which is important to protect the patients.

10.3969/j.issn.2095-4344.2013.24.015

BACKGROUND:In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE:To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS:BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5). bFGF-and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire bal oon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION:BMSCs were successful y transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5%of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P<0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair.

Objective To investigate the role and mechanism of macrophage activation induced by exosomes from high glucose-treated renal tubular epithelial cells.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normal glucose control group (5.5 mmol/L D-glucose) or high glucose group (30.0 mmol/L D-glucose) for 48 h were collected and ultracentrifuged to harvest exosomes.Exosomes were identified by transmission electron microscope and Western blotting.(2) Exosomes were labeled with the green lipophilic fluorescent dye PKH67 and cultured with THP-1 macrophage to investigate whether HK2-derived exosomes could be internalized by THP-1 macrophage.Observing the morphology microscopically and detecting the chemotaxis function of THP-1 macrophages in Transwell chamber after co-cultured with exosomes.The expression of inducible nitric oxide synthase (iNOS),tumor necrosis factor-α (TNF-α),Interleukin-1β (IL-1β),monocyte chemoattractant protein-1 (MCP-1) in cells and supernatants were separately detected by quantitative Real-time PCR (qRT-PCR) and enzyme linked immunosorbent assay (ELISA),and the expression of p-c-Jun NH2-terminal kinase (p-JNK),mitogen-activated protein kinase p-p38 (p-p38MAPK) and nuclear factor κB p65 (NF-κB p65) in THP-1 macrophages were detected by Western blot.Results (1) Vesicles that harvested by ultracentrifugation ranged in size from 30 nm to 100 nm and expressed exosomal marker CD63,TSG101 but absence of calnexin which is a marker of endoplasmic reticulum,suggesting that the exosomes were not contaminated with cells.(2) Results from laser scanning confocal microscope showed that each group of exosomes can be internalized by THP-1 macrophages.Compared with normal glucose exosomes group,high glucose exosomes had increased the expression of iNOS,TNF-α,IL-1β and MCP-1 in THP-1 macrophages (all P ＜ 0.01),moreover,p-JNK,p-p38 MAPK and NF-κB p65 proteins level also increased significantly (all P ＜ 0.01).Conclusions Exosomes from high glucose-treated HK2 cells can induce THP-1 macrophage activation and functional changes through MAPK/NF-κB pathway.