Objective To investigate the effects of adenovirus vectors for BTB and cap'n'collar protein homology 1 (Bach1) small interfering RNA (siRNA) on antioxidanffactors and fibrosis related cytokines in lung fibroblasts (MLF) in transforming growth factor (TGF)-β1 induced mouse.Methods Bach1 siRNA recombinant adenovirus vectors and blankadenovirus vectorwere constructed,then the MLF cells were incubated with TGF-β1 (5 ng/ml) for 24 h and infected with blankvector and successful constructed Bach1 siRNAs.The messenger RNA (mRNA) and protein expression of Bach1,heme oxygenase (HO)-1,glutathione peroxidase 1 (GPx1) and nicotinamide-adenine dinucleotide phosphate:quinone oxidoreductase-1 (NQO1) in cell supernatants were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Changes of fibrosis-related cytokines including TGF-β1 and interleukin (IL)-6 levels in cell supernatants were determined by enzyme-linked immunosorbent assay (ELISA).One-way analysis of variance (ANOVA) was performed for multiple group comparisons and LSD test was used to compare the two groups.Results Bach1 mRNA (2.127±0.089) and protein expression increased significantly after TGF-31 stimulation compared with blank group (1.000±0.067,t=-21.77,P＜0.01),as well as the expression of fibrosis-related cytokines TGF-β1 (52±6) and IL-6 (34±6) in cell supernatants increased significantly after TGF-β1 stimulation compared with blank group (26±4,t=-11.11,P＜0.01 and 20±5,t=-5.32,P＜0.01),but the mRNA expression of HO-1 and GPx1 (0.403±0.040 and 0.567±0.112) also the protein expression decreased significantly compared with mRNA (1.000±0.181,t=25.57,P＜0.01 and 1.000±0.212,t=6.68,P＜0.05) and protein expression in blank group.Follow the Bach1 siRNA treatment,Bach1 mRNA (0.153±0.015) and protein levels were significantly downregulated compared with mRNA (2.129±0.089 and 1.973±0.035,F=1835.95,P＜0.01) and protein expression of TGF-β1 and blank vector group,as well as TGF-β1 (26±3) and IL-6 (11±3) expression in cell supernatant were significantly inhibited compared with TGF-31 (52±6 and 34±6) and blankvector group (49±5 and 33±6) (F=22.25,P＜0.01 and F=28.38,P＜0.01).But the mRNA levels of HO-1 (3.303±0.294) and GPx1 (1.840±0.231) in MLF were promoted significantly compared with TGF-β1 (0.403±0.040 and 0.567±0.112) and blank vector group (0.353±0.057 and 0.667±0.090) (F=53.90,P＜0.01 and F=526.25,P＜0.01).Conclusion Silencing Bach1 rescues TGF-β1 induced reduction of antioxidants and increasethefibrosis in MLF cells.The study offers an experimental basis to explore pathogenesis of oxidative stress and antioxidant therapy for connective tissue disease related inter-stitial lung disease.

Objective To compare the clinical effects of modified single open-door laminoplasty combined with segmental lateral screw fixation with conventional single open-door combined with continuous screw fixation in the treatment of multilevel cervical spondylotic myelopathy (MCSM).Methods Retrospective analysis was used to analyze the fifty-five cases of MCSM who underwent posterior operation in Lianyungang Hospital Affiliated to Xuzhou Medical University from March 2009 to January 2017.Among them,26 cases in the modified group were treated by modified single open-door laminoplasty combined with segmental lateral screw fixation;29 cases in the traditional group were treated by conventional single open-door combined with continuous screw fixation.JOA score collected before the operation and at the last time of the follow-up,the cervical spine mobility,cervical curvature,cervical axial symptom,C5 nerve root palsy and the incidence rate of screw fracture in both groups were compared.Operation time and the intraoperative blood loss were recorded.Results 55 cases were all followed up.The cervical spine mobility at the last time of the follow-up in the modified group was significantly higher than that of the traditional group ((39.63°±5.98°) vs.(36.14°±6.05°),P=0.036).The cervical curvature in the modified group was higher than that of the traditional group ((19.83°±5.61°) vs.(16.02°±7.05°),P=0.030).The operation time and intraoperative blood loss in the modified group were significantly lower than those of the traditional group ((116.3±13.2) min vs.(128.7±14.3) min,(186.7±39.8) ml vs.(228.7±42.5) ml,P=0.002,0.000).In the modified group,the proportion of patients with postoperative cervical axial symptom was 15.38%,the rate of C5 nerve root palsy cases was 8.69%,compared with 48.27% and 34.48%,respectively in the control group,the difference between the two groups was statistically significant (P=0.011,0.022).In the traditional group,2 cases of screw fracture were found at the last follow-up,while there was no such case in the modified group.Conclusion The modified cervical single open-door laminoplasty combined with segmental lateral screw fixation can obtain good effect of nerve decompression,maintain cervical mobility and cervical curvature,reduce the occurrence of postoperative cervical axial symptom,C5 never root palsy,screw fracture and other complications,is a safe and effective surgical method.

Objective To study effect of celecoxib on Wnt/β-catenin signaling pathway and matrix metabolism of chondrocytes in osteoarthritis (OA).Methods Use a modified way of mechanical fixation to make rabbit OA model.Knee OA separated and extractd rabbit chondrocytes were and then culture in medium.The cultured chondroctyes were divided into the Dimethyl sulfoxide (DMSO) control group and the celecoxib group, which were given 1％ DMSO and 10 μmol/L celecoxib respectively.After 48 hours' intervention, the protein expression and gene expression of Wnt/β-catenin signaling pathway related factors, including β-catenin,matrix metalloproteinase (MMP)-3, MMP-13, collagen Ⅱ were detected.T-test was used for the comparison between the two groups.Results Compared with the control group, the levels of type Ⅱ collagen [(275±19) ng/ml vs (202±12) ng/ml, t=-6.21, P＜0.05] and proteoglycan [(58±12) ng/ml vs (30±3) ng/ml, t=-9.23, P＜0.05] significantly increased in celecoxib group (P＜0.05).Meanwhile, compared with the control group, the levels of β-catenin, MMP-3, MMP-13 mRNA and protein in celecoxib group were significantly reduced (P＜0.05).Conclusion In rabbit OA chondrocytes, celecoxib can inhibit Wnt/β-catenin signaling pathway, reduce downstream factors expression including MMPs, thereby increase the cartilage matrix type Ⅱ collagen and proteoglycan levels.

This study revealed that the content of intra and extracellular polysaccharides may be related to the cultural conditions.With decreasing NaHCO 3 or NaNO 3 concentration,the content of intracellular polysaccharide was reduced,Reduction of NaNO 3 concentration still caused the rise of extracellular polysaccharides.The K 2HPO 4 or NaCl concentration also had an effect on the production of both intracellular and extracellular polysaccharides.When PH value of media maintained at 8.0,there was the highest content of intra and extracellular polysaccharides.The same high content also occurred after culture of nine days.In addition the content of intracellular polysaccharide was higher under continuous light than under the light/dark circle.

AIM:To explore the fingerprint method for quality control of Danhong sterile powder for injection (Radix et Rhizoma salviae mihiorrhizae,Flos carthami).METHODS:The samples were determined by Agilent 1100 DAD-HPLC with SinoChrom ODS-BP column(250 mm×4.6 mm,5 μm),by gradient elution using methanol-0.1% phosphoric acid,as mobile phase 30 ℃ column temperature.flow rate of 1.0 mL/min,detection wavelength was set at 280 nm,and inject volume 20 μL.RESULTS:Its fingerprint exhibited better separation for each peak,revealing amount of finger information,Based on the ten batches of Danhong sterile powder for injection,according to the technical requirements of fingerprint on injection of Chinese traditional medicine.CONCLUSION:Proposed HPLC fingerprint can be used as the important evidences for the quality control of Danhong Injection.

AIM:To explore the fingerprint method for quality control of Danhong sterile powder for injection(Radix et Rhizoma salviae miltiorrhizae,Flos carthami).METHODS:The samples were determined by Agilent 1100 DAD-HPLC with SinoChrom ODS-BP column(250 mm?4.6 mm,5 ?m),by gradient elution using methanol-0.1% phosphoric acid,as mobile phase 30 ℃ column temperature,flow rate of 1.0 mL/min,detection wavelength was set at 280 nm,and inject volume 20 ?L.RESULTS:Its fingerprint exhibited better separation for each peak,revealing amount of finger information,Based on the ten batches of Danhong sterile powder for injection,according to the technical requirements of fingerprint on injection of Chinese traditional medicine.CONCLUSION:Proposed HPLC fingerprint can be used as the important evidences for the quality control of Danhong Injection.

Objective To identify the crucial gene implicated in rheumatoid arthritis (RA) pathogenesis by comparing microarray-based gene expression profiles of synovial fibroblast in arthritis patients and that in control.Methods The public datasets were obtained from NCBI GEO and EBI ArrayExpress.The qualified microarray-based gene expression profiles were integrated and normalized using the method implemented in GeneSpring software.Furthermore,the differentially expressed genes (DEGs) were identified using significance analysis of microarrays (SAM) method.The online tool DAVID and STRING were applied to conduct the enrichment analysis and gene product interaction analysis respectively.Results There were two datasets that were qualified and analyzed in the present study.A total of 336 significant DEGs were identified by comparing the whole-genome gene expression profiles from synovial fibroblast of RA patients and control group.Among these DEGs,261 were significantly downregulated and 75 upregulated.About 13.6％ of the downregulated genes were associated with extracellular matrix degradation.The COL9A3 and COL4A5,indispensable component of hyaline cartilage and basement membrane respectively,were significantly downregulated,as well as genes in WNT family,including WNT2,WNT11,and WNT16.In contrast,matrix metalloproteinase 13 (MMP13) was found to be significantly upregulated in RA patients.MMP13 is a matrix metallopeptidase that degrade extracellular matrix and hyaline cartilage,and it could possibly interact with other proteins to regulate morphogenesis.Conclusion Molecular mechanisms underlying RA pathogenesis were investigated by analyzing the public datasets.A few genes that associated with extracellular matrix degradation,construction and regulation,including MMP13,WNT2,WNT11,WNT16,COL9A3 and COL4A5,could be regarded as therapeutic targets in RA treatment.

Objective To explore the relationship between osteopontin (OPN),its downstream product thrombin-cleaved osteopontin (NTOPN) and radiographic changes in osteoarthritis (OA).Methods The levels of OPN and NTOPN in serum and synovial fluid were tested by enzyme-linked immuno sorbent assay (ELISA) kit in 22 OA patients and 21 health controls.The correlation between the above parameters and clinical data such as radiology changes of OA patients was analyzed.Results The serum level of OPN in the OA group was positively correlated with C reactive protein (CRP) (r =0.574,P =0.005).The serum level of OPN in the OA group was slightly lower than that in the healthy control group [(1.8 ± 0.5) μg/L vs (1.7 ± 1.1) μg/L,P ＞ 0.05].Similarly,the serum level of NTOPN in the OA group was significantly higher than that in control group [(21.1 ± 11.7) μg/L vs (14.7 ± 3.6) μg/L,P ＜ 0.05].Meanwhile,serum NTOPN level in female patients was significantly higher than that in male patients [(23.2 ± 11.80) μg/L vs (11.1 ± 4.9) μg/L,P ＜ 0.05].In synovial fluid of OA group,the level of OPN was (7.75 ± 2.33) μg/L,NTOPN was (0.91 ± 0.49) μg/L and the ratio of NTOPN/OPN was 0.12 ± 0.08.The level of OPN of synovial fluid in the OA group was positively correlated with CRP (r =0.574,P =0.005).NTOPN-to-OPN ratio of synovial fluid in the OA group was positively correlated with Kellgren-Lawrence grade (r =0.535,P =0.022).Conclusions The serum level of OPN is correlated with severity of inflammation in the OA patients.Meanwhile,NTOPN-to-OPN ratio of synovial fluid is correlated with OA severity.As a downstream bioactive product of OPN,NTOPN may play an important role in OA pathogenesis.Whether these parameters can be used as predictors of joint lesion severity and inflammatory status is worth further study.

In this study, image analysis technique (IAT) was used in the quantitative analysis of the essential pathological changes in the myocardium. The myocardial samples were taken from 46 patients of rheumatic heart disease, who had undergone valve replacement. Stepwise regression of the data demonstrated myocardial fibrosis and chronic arteriolopathy were remarkably correlated to the heart functions a Her the operation, especially myocardial fibrosis. Discriminants were obtained by stepwise discrimination analysis, and they may be used as predictors.

The distribution of copper-zinc superoxide dismutase (Cu-ZnSOD) in subcellular organelles in 8 cases of normal human myocardium was studied by using immunocytochemistry and immunoelectron microscopic technique and the contents of Cu-Zn SOD in myocardium were measured by radioimmunoassay. The results showed that Cu-ZnSOD was in most myocardial cells. The contents of Cu-ZnSOD in the ventricular myocardial cells were higher than those in the atrial myocardial cells. The Cu-ZnSOD labeled with immunogold particles were distributed throughout cytoplasm and nuclei, but less in mitochondria and a very little in lysosomes. No gold particle was found on the membranous structure of sarcoplasmic reticulum as well as Golgi complex. The results demonstrate that the myocardium was the tissue containing quite abundant Cu-Zn SOD.