Objective To establish a simple, cost-effective method to measure reactive oxygen metabolites ( ROMs) using automatic biochemical analyzer.Methods Serum samples were collected from 50 healthy individuals and 50 hospitalized patientsin Beijing Hospital from September 2013 to November 2013.By setting up and optimizing parameters on automatic biochemical analyzer, a new method of measuring ROMS was established with TBHP as calibrator.130 healthy non-smokers were recruited in Beijing Hospital from November 2013 to December 2013 to establish reference interval, including 73 males and 57 females.The average age was 68.8 ±12.5.According to CLSI documents, there was the establishment of the new method including precision, limit of blank ( LoB ) , limit of detection ( LoD ) , limit of quantitation (LoQ), linearity,interference testing, and reference interval.Results The intra assay imprecision was 1. 5%-4.1% and the total imprecision was 4.4%-9.9%.LoB was 0.85 mmol/L TBHP;LoD was 2. 7mmol/L TBHP;LoQ was 5.9mmol/L TBHP.The linearity was 5.9-600 mmol/L (R2 =0.995).When triglyceride≤28.58 mmol/L, hemoglobin≤173 g/L, vitamin C≤60 mg/dl and bilirubin≤73.8 μmol/L, the deviation was -7.6%, 4.6%, -98.9%, 19.1%respectively.The normal reference interval was 61.9 -88.1 mmol/L TBHP.Conclusions The newly-established method has good performances of precision,LOD and linearity, which can meet the clinical needs.The interference of triglyceride and hemoglobin is small, while vitamin C and bilirubin have stronger interference on measurement.( Chin J Lab Med,2015,38:163-167)

OBJECTIVETo observe the the expression of endoplasmic reticulum stress (ERS) related factors in deep tissue injury (DTI) at pressure ulcer rat and to investigate the ERS mechanism of DTI in muscle tissue and protective effect of 4-phenylbutyric acid (4-PBA) in local tissue.

METHODSFifty male SD rats were randomly devided into control group, model group, experimental group NS group and PBA group, the experimental groups were divided into 4 d, 7 d, 14 d and 21 d group according to the observation time (n = 5). Rats in the PBA group were administrated with gastric perfusion of 4-PBA after the modeling; the NS group was given normal saline of the same quantity. Using HE staining to observe morphologic character. The expression of glucose regulated protein 78 (GRP78), CHOP, Caspase 12 were detected by immunohistochernical staining. Cell apoptosis was detected by TUNEL assay.

RESULTSHE staining results showed that each group demonstrated compression injury compared with control group: cellular swelling, ompaction of nuclear, and apoptosis in muscle tissue. The new muscle fiber in 4-PBA group fused faster than those in NS group. The number of TUNEL positive cells peaked at 4 day after compression, then got decreased on day 7 in muscle tissue, apoptosis positive cells were diminished after 4-PBA treatment. The immunohistochemical staining results showed that the expression of protein GRP78, CHOP, Caspase 12 peakd 4 d after modeling and decreased gradually. The GRP78, CHOP, Caspase 12 protein expression were significantly higher than those of PBA group at all time points (P < 0.05).

CONCLUSIONCell apoptosis induced by endoplasmic reticulum stress took part in deep tissue injury resulting of pressure ulcer, which mechanism might be related to reducing apoptosis mediated by CHOP, Caspase 12.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Male ; Muscle, Skeletal ; pathology ; Phenylbutyrates ; pharmacology ; Pressure Ulcer ; physiopathology ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Transcription Factor CHOP ; metabolism

OBJECTIVETo observe effects of serum Bushen Huoxue prescription(Chinese characters) on classic Wnt/β-catenin signaling pathways of osteoblasts, and explore mechanism of Bushen Huoxue prescription (Chinese characters) for preventing osteoporosis.

METHODSTwenty health female rats were randomly divided into two groups, including Bushen Huoxue (Chinese characters) group and saline group,10 in each group. Bushen Huoxue (Chinese characters) group and Saline group were gavaged Bushen Huoxue and saline every day for 1 week. Bushenhuoxue containing serum and saline containing serum were got according to methods of serum preparation of drug-containing. The osteoblasts was cultured with neonatal rat skull according to Enzyme Consumer Law, and was identified by Wright-Giemsa staining (R-J) and alkaline phosphatase staining (ALP). The third generation of osteoblasts was divided into three groups, including saline group, normal group,Bushen Huoxue (Chinese characters) group. Each group were added to 15% appropriate medium. ALP activity of osteoblasts and osteoblasts proliferation rate were tested, mineralized nodules were observed, the expression of β-catenin, Runx2, Osx mRNA of osteoblasts were tested by RT-PCR.

RESULTSThere was blue granules in cytoplasm, cell nucleus was flint with 1 to 3 nucleoli showed by R-J staining, morphology of osteoblasts were cultured. ALP staining showed cytoplasm with purple granules, the results showed that the cultured cell was osteoblasts. The content of ALP in Bushen Huoxue (Chinese characters)group was (6.272±0.131) ,appreciation rate was (0.81? 0.172), and could significantly improve differentiation and proliferation activity of osteoblasts compared with Saline group (P< 0.01). There were four different size orange nodules, the Maximun nodule was 1.0 x 1.0 cm in Bushen Huoxue (Chinese characters) group after Alizarin red staining, the results showed Bushen Huoxue (Chinese characters)group could obviously improve mineralization of osteoblasts. The expression of mRNA of β-catenin, Runx2 and Osx in Bushen Huoxue (Chinese characters)group were (1.782±0.944), (1.935±0.994) and (1.610±0.811) by RT-PCR,it was significantly increased compared with saline group (P<0.01), but there was no difference between Bushen Huoxue (Chinese characters)group and normal group (P>0.05).

CONCLUSIONBushen Huoxue (Chinese characters)group could obviously promote differentiation, proliferation and mineralization of osteoblasts through activation of Wnt, β-catenin signaling pathway. It suggested that the mechanism of action of Bushen Huoxue (O'f f Il.t)particle clould prevent osteoporosis through the activation of Wnt, β-catenin signaling pathway.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteoporosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Wnt Proteins ; genetics ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; genetics ; metabolism

Chronic osteomyelitis is one of the most common disorder in clinic. In recent years due to diabetes, peripheral vascular disease and trauma induced disease increased, the prevalence rate increased. With the development of magnetic resonance imaging and CT imaging technology, it greatly improved the accuracy of clinical diagnosis of chronic osteomyclitis and ability to describe the infection characteristics, and provide a reliable basis for clinical treatment. The current research on chronic osteomyelitis mainly concentrated on the aspects of imaging applications and ways of using antibiotic optimization control inflammation, defect restoration and reconstruction of blood supply and treatment. But the best time to the antibiotic therapy and the use of program is still uncertain, for after debridement, bone grafting time and defect repair function of fast recovery still need further research.
Anti-Bacterial Agents ; therapeutic use ; Chronic Disease ; Humans ; Osteomyelitis ; diagnosis ; therapy

[ ABSTRACT] AIM:To explore the effects of hydrogen sulfide ( H2 S) on the myocardial fibrosis in a rat model of diabetes and its mechanism.METHODS:Single intraperitoneal injection of streptozotocin ( STZ) was utilized to establish a rat model of diabetes.Sodium hydrosulfide was used as an exogenous donor of hydrogen sulfide.Male SD rats were ran-domly divided into control group, STZ group, STZ+H2 S group and H2 S group.Eight weeks later, HE and VG staining methods were used to observe the collagen distribution and collagen volume fraction was measured by image analysis.The expression levels of type I collagen, PPARγand NF-κB in the cardiac tissues were determined by Western blotting.RE-SULTS:Compared with control group, collagen distribution and the expression levels of type I collagen and NF-κB in the cardiac tissues were markedly increased (P<0.05), while PPARγwas significantly decreased in STZ group (P<0.05), but these indexes were reversed significantly in STZ+H2S group (P<0.05).The expression levels of type I collagen, PPARγand NF-κB had no significant difference between H2 S group and control group.CONCLUSION:Hydrogen sulfide attenuates cardiac fibrosis in diabetic rats, and its mechanism may be related to PPARγ-NF-κB signaling pathway.

Objective To investigate the influences of UVA on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells induced by interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α). Methods HaCaT cells were cultured in the presence of IFN-7 and TNF-a and irradiated with UVA of 2, 4 and 8 J/cm~2, respectively; those cells receiving neither treatment with IFN-γ or TNF-α nor UVA irradiation served as the negative control, and those receiving only cytokine treatment but no irradiation as the positive control. After another 24-hour culture, enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein levels of CXCL11/I-TAC in the supernatant of HaCaT celb, real time PCR to measure the mRNA expression of CXCL11/I-TAC in these HaCaT cells. Results As far as the negative control HaCaT cells were concerned, there was a minor secretion of CXCL11/I-TAC protein and expression of CXCL11/I-TAC mRNA. After treatment with IFN-7 and TNF-a of 10 μg/L, the protein and mRNA expressions of CXCL11/ I-TAC were synergistically upregulated, whereas the induced secretion and expression of CXCL11/I-TAC by HaCaT cells were dose-dependently inhibited by UVA irradiation. Conclusions UVA irradiation inhibits the secretion and expression of CXCL11/I-TAC by HaCaT cells, which in turn suppresses the chemotaxis of Th1/ Tel cells in some degree.

Objective:To investigate the expression of Th1 chemokine CXCL9,CXCL10,CXCL11,Th2 chemokine CCL22 and their receptors in the lesions of bullous pemphigoid (BP).Methods:Immunohistochemical assay was performed to detect the expression of CXCL9,CXCL10,CXCL11,CCL22 and their receptors CXCR3 and CCR4 in BP lesions and normal control skin.Results:CXCL9,CXCL10,CXCL11,CCL22,CXCR3 and CCR4 were overexpressed in BP lesions than those in normal control skin (P＜0.01).The positive rates of CXCL9,CXCL10,CXCL11 and CXCR3 in BP lesions were 50%(15/30),46.7%(14/30),46.7%(14/30) and 53.3%(16/30),respectively.The positive rates of CCL22 and CCR4 were 66.7% (20/30) and 56.7% (17/30).Conclusion:The overexpression of Th1 chemokine CXCL9,CXCL10,CXCL11,Th2 chemokine CCL22 and their receptors may play important roles in the pathogenesis of BP.

Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

ObjectiveTo evaluate the diagnostic performance of CTCA in predicting myocardial perfusion defects through comparative analysis between MPI defects and severity of coronary stenosis on CTCA.MethodsFour hundred and seventy-eight patients who underwent CTCA and 99Tcm-MIBI MPI in the same period were analyzed retrospectively.According to the severity of coronary stenosis judged by visual evaluation of the vessel diameter,the patients were divided into five groups:no stenosis,mild stenosis,moderate stenosis,severe stenosis and total occlusion.MPI results were classified as negative or positive for perfusion defects,and the prevalence of perfusion defects in every group was calculated per-patient and per-vessel basis.The cut-off of stenotic severity for predicting myocardial perfusion defects was designated as 50％ or 75％,with MPI as standard reference.True positive,true negative,false positive and false negative statistics were thus determined separately on patient and vessel basis.The diagnostic performance for CTCA were calculated and compared.Pearson Chi-square and its partition tests or Fisher exact test were used to compare ordinal variables.ResultsFifty-eight patients showed myocardial perfusion defects.Either by patientbased or vessel-based analysis,the prevalence of myocardial perfusion defects showed an increasing trend with greater coronary artery stenosis in each group,and there were statistical differences among them (x2 =116.62 and 483.83,both P ＜ 0.05).On patient-based analysis,sensitivity ( SN),specificity ( SP),positive predictive value( PPV),negative predictive value(NPV) and accuracy (AC) for CTCA predicting myocardial perfusion defects were 62.1 ％ ( 36/58 ) and 34.5％ ( 20/58 ) (x2 =8.84,P ＜ 0.05 ),84.5％(355/420) and 97.1％ (408/420) (x2 =40.16,P ＜0.05),35.6％ (30/101) and 62.5％ (20/32) (x2 =7.19,P＜0.05),94.2％ (355/377) and 91.5％ (408/446) (x2 =2.18,P ＞0.05),81.8％ (391/478)and 89.5 ％ (428/478) (x2 =11.66,P ＜ 0.05 ) when the cutoff was set to 50％ and 75％,respectively.On vessel-based analysis,the SN,SP,PPV,NPV and AC for CTCA predicting myocardial perfusion defects were 58.8％ (40/68) and 30.9％ (21/68) (x2 =10.73,P ＜ 0.05),95.9％ (1768/1844) and 99.0％ (1826/1844) (x2 =36.72,P ＜ 0.05 ),34.5％ (40/116) and 53.8％ (21/39) (x2 =4.59,P ＜0.05 ),98.4％ (1768/1796) and 97.5％ ( 1826/1873 ) (x2 =4.14,P ＜ 0.05 ),94.6％ ( 1808/1912 ) and 96.6％ ( 1847/1912 ) (x2 =10.31,P ＜ 0.05 ),respectively.ConclusionsThe prevalence of myocardial perfusion defects correlates positively with the severity of coronary stenosis seen on CTCA.CTCA may predict perfusion defects with high SP and NPV.However,the PPV of CTCA in predicting myocardial perfusion defects is poor when the stenosis cut-off is set at 50％.It is significantly improved when the cutoff value is set at 75 ％.

Objective: To investigate the impact and its possible mechanism of hydrogen sulfide (H2S) on myocardial collagen remodeling in experimental rats with diabetic mellitus (DM).
Methods: Rat’s DM model was established by intraperitoneal injection of STZ at 40 mg/kg. A total of 40 SD rats were randomly divided into 4 groups:Control group, DM group, DM+NaHS group, in which NaHS worked as exogenous donor of H2S and NaHS control group. n=10 in each group, all animals were treated for 8 weeks. The cardiac collagen deposition was observed by Masson staining, protein expressions of cardiac collagen types I, III, IV and transforming growth factorβ1 (TGF-β1), connective tissue growth factor (CTGF) were examined by Western blot analysis.
Results: Compared with Control group, DM group showed increased protein expressions of cardiac collagen types I and III, up-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups. Compared with DM group, DM+NaHS group presented reduced cardiac collagen expression, decreased expression of collagen types I and III, down-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups.
Conclusion: H2S may improve the myocardial collagen remodeling in experimental DM rats, the mechanism might be related to the down-regulation of TGF-β1 and CTGF expression.