Objective To investigate the expression of aquaporin-4 (AQP4) in contused brain and contralateral side and correlation of AQP4 level with brain edema.Methods A total of 70 health adult SD rats were divided into sham operation group (n =10) and brain contusion group (n =60) according to the random number table.Except for no impact in sham operation group,the remaining operations were the same in both groups.Ten rats in brain contusion group were separately sacrificed at 1,6,24,48 and 72 hours,and 7 days.Pathological changes in brain tissues of rats were detected and immunohistochemistry staining and Western-blot were used to measure AQP-4 expression.Results There were no abnormalities of brain tissue structure in sham operation group with AQP4 expressed mainly in glial cells and vascular endothelial cells.With respect to the injured side in brain contusion group,mixed brain edema that consisted of angioedema and cellular edema occurred along with tissue necrosis,massive inflammatory cell infiltration,and microgial proliferation.AQP4 was apparently expressed in gliocytes and vascular endothelial cells,which showed a fall at 1 hour,the lowest at 6 hours,the peak at 24 hours,slow fall at 48 hours,a second peak at 72 hours,and then a fall until 7 days.Level of AQP4 other than that at 1 hour had significant difference compared with sham operation group (P ＜ 0.01).Whereas the contralateral side in brain contusion group showed no pathological abnormalities at 1 hour,but cellular edema emerged at 6 hours followed by mixed brain edema mainly characterized as cellular edema at 24 hours,continued cellular edema but alleviated angioedema at 48 and 72 hours,and alleviated cellular edema at day 7.AQP4 was up-regulated in gliocytes and vascular endothelial cells,which decreased at 1 hour (1.313 ± 0.01),minimized at 6 hours (0.922 ± 0.03),peaked at 24 hours (2.848 ± 0.020),fell at 48 hours,peaked again at 72 hours (2.662 ± 0.02) and fell to almost normal level at day 7.By contrast with sham operation group,level of AQP4 had significant difference other than that at 1 hour and 7 days (P ＜ 0.01).Conclusions After brain contusion,angioedema and subsequent cellular edema emerge in the contused side.Pathological changes are delayed in non-contused side with cellular edema ahead of angioedema.Level of AQP4 is closely related with traumatic brain edema.

OBJECTIVEThis study aims to test the accuracy and precision of iWitness photogrammetry for measuring the facial tissues of mannequin head.

METHODSUnder ideal circumstances, the 3D landmark coordinates were repeatedly obtained from a mannequin head using iWitness photogrammetric system with different parameters, to examine the precision of this system. The differences between the 3D data and their true distance values of mannequin head were computed.

RESULTSOperator error of 3D system in non-zoom and zoom status were 0.20 mm and 0.09 mm, and the difference was significant (P 0.05). Image captured error of 3D system was 0.283 mm, and there was no significant difference compared with the same group of images (P>0.05). Error of 3D systen with recalibration was 0.251 mm, and the difference was not statistically significant compared with image captured error (P>0.05). Good congruence was observed between means derived from the 3D photos and direct anthropometry, with difference ranging from -0.4 mm to +0.4 mm.

CONCLUSIONThis study provides further evidence of the high reliability of iWitness photogrammetry for several craniofacial measurements, including landmarks and inter-landmark distances. The evaluated system can be recommended for the evaluation and documentation of the facial surface.

BACKGROUND:Our previous studies have shown that strontium-doped calcium polyphosphate containing low-dose strontium appears to have a significant effect on angiogenesis-related behaviors of monocultured umbilical vein endothelial cells and osteoblasts.
OBJECTIVE:To investigate the effect of strontium-doped calcium polyphosphate on angiogenesis-related behaviors of umbilical vein endothelial cells and osteoblasts co-cultured, including celladhesion, spreading, proliferation, as wel as the protein secretion of vascular endothelial growth factor and basic fibroblast growth factor from co-culture system in vitro.
METHODS:Human umbilical vein endothelial cells and osteoblastic cells (MG63) were utilized in this study. cells from passage 3 were used for preparation of the cel-scaffold constructs. After placed in 24-wel plate at a ratio of 2:1, human umbilical vein endothelial cells and MG63 cells were seeded onto strontium-doped calcium
polyphosphate, calcium polyphosphate and hydroxyapatite scaffolds and co-cultured for 7 days. The vascular endothelial growth factor and basic fibroblast growth factor protein levels were determined through a double ligand enzyme-linked immunosorbent assay. The colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay was performed to quantify the effect of scaffolds on cellproliferation.
RESULTS AND CONCLUSION:Compared with those on calcium polyphosphate and hydroxyapatite scaffolds, cells on strontium-doped calcium polyphosphate scaffolds attached and spread better with a significantly improved cellproliferation. More importantly, the vascular endothelial growth factor and basic fibroblast growth factor expressions were significantly higher in the strontium-doped calcium polyphosphate group than the other two groups (P<0.05), indicating strontium-doped calcium polyphosphate can up-regulate levels of vascular endothelial growth factor and basic fibroblast growth factor proteins.

AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .

OBJECTIVE To investigate the molecular mechanisms of resveratrol( Res)in renal interstitial fibrosis(RlF)in rats with unilateral ureteral obstruction(UUO). METHODS Forty-eight Spra-gur-Dawley rats were randomly divided into UUO( normal saline,n = 16),UUO with Res treatment (Res,20 mg·kg-1 ,n=16),and sham-operation(sham,n=16)models. The kidneys were excised on the 7th and 14th day. The deposition of collagen fiber in the kidney was detected with HE and Masson staining. The levels of sonic hedgehog(SHH,an inducer of SHH pathway)in kidney tissues were deter-mined by ELlSA. lmmunohistochemical analysis was performed to evaluate the protein expression of SHH signaling-related molecules,including SHH,smoothened(Smo),patched-1(Ptch1),and Gli1, proliferating cell nuclear antigen(PCNA)and matrix component typeⅢ collagen. The mRNA expression levels of Smo,Ptch1 and Gli1 were detected by real-time RT-PCR. RESULTS The degree of RlF observed with HE and Masson staining was obviously increased in UUO kidneys,but decreased in Res-treated kidneys. Enhanced expression levels of typeⅢ collagen and PCNA in UUO rats were suppressed by Res treatment(P﹤0.05). Res administration decreased the expression levels of SHH,Smo,and Gli1 (P﹤0.05),but increased the expression of Ptch1(P﹤0.05),suggesting that Res inhibit the obstruction-induced activation of SHH signaling. CONCLUSION Res can attenuate RlF in UUO rats,and the possi-ble mechanism is that Res down-regulates the activity of SHH signaling and inhibits cellular proliferation, resulting in inhibition of matrix accumulation in renal interstitium of UUO rats.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Objective:A method that is based on microfluidic cell chip technology was developed for the first time to analyze CD14+monocyte myeloperoxidase (MPO) expression in myelomonocytic leukemia (M4) patients. CD14+monocyte MPO expression in M4 patients was preliminarily discussed. Methods:a. The chip was prepared by using polydimethylsiloxane as the host material and by secondary foam molding. b. A total of 48 clinically diagnosed M4 patients and 52 patients with normal myelogram were included as the test and control groups, respectively. c. A method based on the microfluidic cell chip approach was established to detect CD14+mono-cytes and to determine the positive rate and degree of MPO expression in the cells. d. The microfluidic cell chip technique was used to compare CD14+monocyte MPO expression in M4 patients with that in the control. Results:a. The designed microfluidic single cell analysis chip allowed the entry of granulocytes into the corresponding microfluidic channels. Thus, blood cells were separated. Numer-ous ghost corpuscles surrounded the separated white blood cells (WBCs). WBC morphology did not show obvious changes. b. The posi-tive rate of MPO expression and the activity of CD14+monocytes in the bone marrow of M4 patients were significantly higher than those in the bone marrow of the control (P<0.05). Conclusion:A method based on microfluidic single cell technology was developed for the first time to analyze the MPO expression in CD14+monocytes. CD14+monocyte MPO activity in M4 patients was significantly higher than in the control. CD14+monocyte MPO activity can be used as an auxiliary examination marker for clinical diagnosis.

Broth microdilution method was hired to measure the minimum inhibitory concentrations (MICs) of luteolin against Trueperella pyogenes,Escherichia coli,Salmonella and Streptococcus in order to evaluate the antimicrobial activity of luteolin in vitro.Meanwhile,the bacteria growth curves in medium containing sub-inhibitory concentration of luteolin were measured in this test.The results indicated that Tureperella pyogenes was the most sensitive to luteolin with a MIC of 0.078 g/L than that of these strains;and Salmonella was also sensitive to luteolin (MIC:1.25 g/L).However,The inhibitory effect of luteolin on Escherichia coli and Streptococcus is relatively weak,and shared the same MIC with 2.5 g/L.Luteolin showed inhibitory effects on the growth curves of all the strains in this test at sub-inhibitory concentration,and the inhibitory effects on the growth curves increased with the concentration of luteolin(P＜0.05).

Objective To investigate the correlation between serum amyloid A (SAA) and disease activity (DAS28) in patients with rheumatoid arthritis (RA). Methods Forty-four patients with RA, 35 patients with systemic lupus erythe-matosus (SLE), 18 patients with osteoarthritis (OA) and 30 healthy controls (HC) were enrolled in this study. The levels of SAA were measured by ELISA. Erythrocyte sedimentation rate (ESR) was measured by the Westergren method. The value of serum C reactive protein (CRP) was examined by immunonephelometry assay. The correlation between SAA and DAS 28, ESR and CRP was assessed, respectively. Results The SAA levels were significantly higher in RA group than those of SLE, OA, and HC groups (P<0.05). The serum ESR and CRP levels were both higher in RA group than those of OA and HC groups (P>0.05), but there was no significant difference between RA group and SLE group. There was positive correlation between SAA and DAS28, ESR, and CRP levels (rs=0.790, P<0.001;rs=0.674, P<0.001;rs=0.679, P=0.004), respective-ly. Conclusion SAA may be a new serological marker to assess disease activity in RA.

Objective To investigate the clinical characteristics and methods of diagnosis and treatment of granular lymphocytic leukemia (LGLL).Methods Clinical data of 3 patients with LGLL were retrospectively analyzed and relevant literature was reviewed.Results 3 patients were all onset with lymphocytosis,whose conditions progressed slowly.The diagnosis of 2 patients was T-LGLL with immunological characteristics of CD3+ CD4 CD8+ CD56-CD57+.The other patient' s diagnosis was NK-LGLL,whose immunological characteristic was CD3-CD4-CD8-CD56+ CD57-.Two of them didn' t need any treatment.One of them was treated with cyclosporine because of agranulocytosis and recurrent infection.Conclusions LGLL is a group of heterogeneous diseases,which clinical characteristic and prognosis are different.Flow cytometric immunopheotype,TCR Vβ analysis and TCR gene rearrangement are helpful to diagnosis.