Objective To observe the effect of ethanol extract and ethyl acetate extract of Capparis Spinosa on the thickness of dermis,synthesis of collagen type Ⅰ,type Ⅲ,and expression of transforming growth factor-β1 in mouse models of scleroderma.Methods Mouse models of scleroderma were established through local injection of bleomycin on the back once a day for 4 weeks.After confirmation of model establishment,72 mouse models were equally and randomly divided into three groups.Two groups received topical treatment with ethanol extract of Capparis Spinosa and ethyl acetate extract of Capparis Spinosa,respectively,no treatment was given to the rest of the control group.After 2-,4-,6-week treatment,8 mice were sacrificed and tissue samples were obtained from the back,and subiected to the measurement of dermal thickness by HE staining,as well as to the analysis of expression of collagen type Ⅰ,collagen type Ⅲ and transforming growth factor-β1 by immunohistochemical staining.Results On week 2,4,6,the thickness of dermis was 23.22,24.94,19.97 μm respectively in mice treated with ethanol extract of Capparis Spinosa,27.66.26.15,22.13 μm respectively in those treated with ethyl acetate extract of Capparis Spinosa.Compared with the mouse models without treatment,the thickness of dermis significantly decreased(F=12.99,P＜0.01),the expression of collagen type Ⅰ(F=7.47,P＜0.01)and transforming growth factor-β1(F=11.76,P＜0.01)were also inhibited in those receiving treatment.However,the expression of collagen type Ⅲ was not affected obviously by the treatment.Conclusion The ethanol extract and ethyl acetate extract of Capparis Spinosa have the effect against skin fibrosis.

ObjectiveTo study the state of oxidative injury induced by sodium arsenite(NaAsO2) in SV-40-immortalized normal uroepithelial (SV-HUC-1 ) cells.Methods SV-HUC-1 cells were exposed to different concentrations of NaAsO2[0(control),1,2,4,8,10 μmol/L] for 24 h,intracellular reactive oxygen species (ROS) was determined by flow cytometry,and the content ofintracellular nitrotyrosine(NT) and the 8-Hydroxydeoxyguanosine (8-OHdG) levels of cell culture medium were detected by enzyme linked immunosorbent assay (ELISA).Results After 24 h treatment,ROS levels(81.76 ± 4.91,95.23 ± 2.17,126.61 ± 17.95,126.74 ± 27.77,114.18 ± 9.65) of SV-HUC-1 cells in the 1,2,4,8,10 μmol/L NaAsO2 exposure groups were significantly higher than those of the control group (69.84 ± 1.28,P ＜ 0.05 or ＜ 0.01 ),ROS levels and exposure dose were positively correlated significantly(r =0.818,P＜ 0.01); the content of NT in the 10 μmol/L NaAsO2 exposure group[(919.66 ± 206.33) μg/L] was significantly higher than that in the control group[ (238.19 ± 38.28)μg/L,P ＜ 0.01 ],NT content and dye concentrations of arsenic also had dose-response relationship (r =0.617,P ＜ 0.01); after 24 h the cells were treated with arsenic,no significant difference of 8-OHdG content in the culture medium was observed(F =2.127,P ＞ 0.05 ).ConclusionNaAsO2 can cause SV-HUC-1 cell oxidative damage.

OBJECTIVETo analyze the data of angle variation on traction based on a finite element model of complete cervical spine with straight physiological curvature, and try to give experimental reference and suggestion in treating cervical spondylosis.

METHODSA 43-year-old female patient with straight cervical spine was chosen and the CT scan data were collected. By using specially designed modeling system, a high quality finite element model of complete cervical spine with straight physiological curvature is generated,which included ligament and muscle according to anatomy. After the model was confirmed, traction was loaded with angle 0 degree, anterior 5 degrees, 10 degrees, 15 degrees, 20 degrees, 25 degrees, to observe the data of distance change on between adjacent intervertebral foramen, processus articularis, uncovertedral joint, intervertebral discs, and stress of anulus fibrosus and nucleus pulposus.

RESULTSWhen the angle was 0 degrees-15 degrees, the distance between intervertebral foramen, Luschka joint and processus articularis posterioris was enlarged, the tensile stress was adequate and compressive stress was small. It met the clinical requests.

CONCLUSION0 degree-15 degrees anterior position is suggested for the treatment of cervical spondylosis.

Adult ; Biomechanical Phenomena ; Cervical Vertebrae ; anatomy & histology ; surgery ; Female ; Finite Element Analysis ; Humans ; Spondylosis ; surgery ; Traction ; methods

OBJECTIVETo compare the clinical effects between hip anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery and posterior K-L approach combined with quadratus femoris bone flap transplantation for the treatment of femoral neck fracture of Garden III-IV in young and middle-aged patients.

METHODSFrom January 2004 to January 2011,46 patients with femoral neck fractures were treated by two kinds of operation. Among them, 20 cases were treated with anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery, included 12 males and 8 females with an average age of (32.1 ± 7.3) years old, involved 12 cases of Garden III and 8 cases of Garden IV. The other 26 cases were treated with posterior K-L approach combined with quadratus femoris bone flap transplantation, included 20 males and 6 females with an average age of (37.8 ± 6.9) years old, involved 16 cases of Garden III and 10 cases of Garden IV. The index of hospitalization (hospitalization time, total cost, operative time, intraoperative blood loss, postoperative complications), the quality index of operation (fracture reduction, position of internal fixation, fracture healing time, nonunion and femoral head necrosis) of two groups were observed and compared. Hip joint function were evaluated by Harris score.

RESULTSAll patients were followed up from 28 to 41 months with an average of 36 months. The intraoperative blood loss of group S-P (92.3 ± 10.4) ml was less than that of group K-L (132.4 ± 11.2) ml, there was significant difference between two groups (P < 0.05). The operation time of group S-P (81.4 ± 9.2) min was more than that of group K-L (67.1 ± 4.5) min, the difference was statistically significant (P < 0.05). One case in group S-P and 9 cases in group K-L appeared postoperative complications, there was significant difference between two groups (P < 0.05). The fracture healing time of S-P group (83.5 ± 7.3) d was shorter than that of group K-L (103.2 ± 12.6) d, there was significant difference between two groups (P < 0.05). At 30 months after operation, there were significant difference in Harris scoring between two groups (P < 0.05).

CONCLUSIONAnterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery for treatment of femoral neck fracture of Garden III-IV of young and middle-aged patients, it has characteristics in clear anatomic and easy to operate. As compared with K-L approach, S-P approach can better reserve residual blood supply of femoral neck. While combining with the iliac bone flap transplantation with deep circumflex iliac artery, it could better reconstruct the blood supply of femoral neck to promote fracture healing.

Adult ; Case-Control Studies ; Female ; Femoral Neck Fractures ; physiopathology ; surgery ; Fracture Healing ; Humans ; Iliac Artery ; Male ; Middle Aged ; Surgical Flaps ; transplantation

There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure

Objective To construct a human phage single chain-antibody library,and to sieve out the antibody ScFv against lung cancer from the library.Methods Total RNA was abstracted from lymph node tissue of the lung cancer,and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined by a DNA linker by SOE-PCR to form the single chain variable fragment (ScFv) gene. ScFv gene was coloned into the phage vector pCANTAB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression.Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning,the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis.Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.

ABSTRACT:Objective To investigate the effect and mechanism of Hedgehog signaling pathway on the invasion of breast cancer cells in vitro.Methods The SHH,SMO and Gli-1 expression levels of breast cancer cell line MDA-231 and normal mammary epithelial cell line MCF-10A were detected by Western blot at protein level and by Real-time RT-PCR at mRNA level.Next,shRNA vector was transfected into the MDA-2 3 1 cells with highly expressed SMO,and the stable transfected cells were selected by G4 1 8 .Western blot and Real-time RT-PCR were performed to observe the inhibitory effect of RNAi on SMO expression.MTT assay was used to assess the influence of SMO siRNA on cell proliferation.Transwell assay was applied to observe cell invasion ability.The expressions of Gli-1,Snail,MMP-9,E-cadherin and Vimentin protein were determined by Western blot.Results Compared with those of normal mammary epithelial cell line MCF-10A,the expressions of SHH,SMO and Gli-1 were significantly increased.The invasion of MDA-2 3 1 cells was inhibited significantly after SMO silencing.Additionally, the protein expressions of Gli-1 , Snail, MMP-9 and Vimentin were obviously inhibited, and E-cadherin was significantly increased.Conclusion Mutative activation of Hedgehog signaling pathway in breast cancer cells promotes cell invasion probably through induction of epithelial-mesenchymal transition of the tumor cells.

Objective To investigate the inhibitory effect of matrine on inflammation by regulating Th1/Th2 bal-ance in asthmatic rats and the underlying mechanism related to SOCS3.Methods Ovalbumin-sensitized rats were established as asthma model, Animals randomly divided into four groups, as follows: control ( without any treatment) , model group, treatment group A ( low-dose matrine treated asthma rats ) and treatment group B ( high-dose matrine treated asthma rats) .The eosinophil counting, goblet cells percentage, inflammatory cell in-filtration in rat lung were analyzed and scored by morphological examination .IL-4 and IFN-γlevel in BALF were determined by ELISA and IFN-γ/IL-4 ratio was further calculated.Furthermore, the expression of SOCS3 in mRNA and protein level were detected by qRT-PCR and Western blot, respectively.Results Eosinophil count and percentage, goblet cell percentage and inflammatory cell infiltration score were significantly lower than that in treatment group A and B as compare to model group ( P<0.05 ) .The group A exhibited a lower IFN-γlevel and a higher IL-4 level ( P<0.05 ) .IFN-γlevel in treatment group A and B were higher while IL-4 level were lower as compare to model group.Meanwhile, SOCS3 mRNA level in rat lung tissue was elevated in model group.Ma-trine treatment decreased SOCS3 expression in group A and B .Similar trend was found in SOCS3 protein level. Conclusions Matrine may exhibit antiinflammatory effect by inhibiting SOCS3 expression and correcting Th1/Th2 balance in asthmatic rats.

Objective To investigate the dose-effect relationship of mRNA level of sensitive mitochondrial genes in human lymphoblastoid cells induced by ionizing radiation.Methods Seven human sensitive genes,including ND3,Cyt b,COX Ⅰ,COX Ⅱ,COX Ⅲ,ATPase6 and ATPase8 were chosen.Changes of mRNA level of these genes were detected by RT-PCR and Real-Time PCR at 24 h after irradiation in human lymphoblastoid cells,which were exposed to 0 - 15 Gy of 60 Co γ-rays.Results The expression of these 7 genes at mRNA level was up-regulated 24 h after irradiation in human lymphoblastoid cells.The level of gene expression of COX Ⅰ,which belongs to complex Ⅳ of mitochondrial respiratory chain,was most obvious,and the peak occurred after irradiation of 8 Gy,which was 13 times of the control group.A good dose-effect relationship was showed for COX Ⅲ gene expression at dose range of 3 -10 Gy as well as 3 - 15 Gy for other 3 genes including ND3,ATPase6 and ATPase8.Conclusions Gene expression levels of COX Ⅲ,ND3,ATPase6 and ATPase8 24h post-irradiation at certain irradiation dose range could be used for radiation damage biomarkers.

Objective To construct a human phage single chain-antibody library, and to sieve out the antibody ScFv against lung cancer from the library. Methods Total RNA was abstracted from lymph node tissue of the lung cancer, and was used to amplify V_H and V_L gene by RT-PCR. V_H and V_L were joined by a DNA linker by SOE-PCR to form the single chain variable fragment ( ScFv) gene. ScFv gene was coloned into the phage vector pCANT-AB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.