Background Epithelial-mesenchymal transition (EMT) is a major event in the pathogenesis of posterior capsular opacification (PCO),and the expressions of α-smooth muscle actin (α-SMA) is the marker of EMT.Previous studies showed that breviscapine plays an important role in anti-fibrosis and suppression EMT,however,the mechanism of its effect on EMT in LECs is unclear.Objective Present study was to investigate the effect of breviscapine on the expression of α-SMA and fibronectin (FN) in human LECs induced by transforming growth factor-β2 (TGF-β2).Methods Human LECs strain,HLE-B3,was cultured and passaged in DMEM containing 10％ fetal bovine serum.Different concentrations of breviscapine (6.75,12.75,25.00,50.00 and 100.00 mg/L)were added into the medium for 24,48 and 72 hours respectively,and then cell cunting kit-8 (CCK-8) was used to evaluate the half maximal inhibitory concentration (IC50) of breviscapine to HLE-B3.In addition,HLE-B3 was subcultured at the density of 1 × 106/hole and divided into 4 groups.The cells were in free-serum medium as the normal control;the cells were exposed in 10 μg/L TGF-β2 as TGF-β2 group;while in the breviscapine group,10 mg/L of breviscapine was added into the culture medium and another group was the combination of 10 mg/L breviscapine and 10 μg/L of TGF-β2 treatment.Real-time PCR and Western blot were used to detect the mRNA expressions of α-SMA and FN as well as their protein in HLE-B3 72 hours after cultured.Results The inhibitory rate of breviscapine to HLE-B3 proliferation was gradually elevated with the increase of concentration of breviscapine,showing a significant inhibition of the cell proliferation among the different groups and at various time points (F =292.851,P=0.000;F =65.037,P=0.000).IC50 of HLE-B3 at 72 hours was 22 mg/L,and therefore,in rest of experiment 10 mg/L of breviscapine was used which was 1/2 only half of the IC50.α-SMA and FN were expressed in cultured normal HLE-B3.The expressing level of α-SMA mRNA and FN mRNA in the HLE-B3 was significantly different among the normal group,TGF-β22 group,breviscapine treatment group and combination group as well as at various time points (F =105.490,P =0.000 ; F =1041.414,P =0.000).Similarly,the protein expressions of α-SM A and FN in the HLE-B3 was significantly different among the four groups and different time points (F=136.872,P=0.000;F=119.820,P=0.000).The expression levels of α-SMA and FN mRNA and their proteins in HLE-B3 were remarkably increased in the 10 μg/L TGF-β2 group compared with the normal control group (at all P=0.000),and those in the combination group were obviously declined in comparison to the TGF-β2 group (P =0.001,0.001,0.001,0.010).No significant difference was found in the expressions of α-SMA and FN in the HLE-B3 between the breviscapine group and normal control group in both transcriptional level and protein level (P =0.551,0.292,0.551,0.360).Conclusions 10 mg/L breviscapine can arrest the proliferation and EMT of human LECs.This result suggests that using breviscapine may be a potential prophylactic approach in the prevention of PCO.

Reactive oxygen species (ROS) is a kind of molecules derived by oxygen in the metabolic process of aerobic cells, which mainly includes superoxide, hydroxyl radicals, alkoxyl, hydrogen peroxide, hypochlorous acid, ozone, etc. They can destroy the structure and function of cells through the damage of biological macromolecules such as DNA, proteins and the lipid peroxidation. ROS also can regulate the proliferation, differentiation and apoptosis of cells through several signaling pathways and participate in fibrogenesis of many organs including hepatic and pulmonary fibrosis. Recent study shows that ROS might have an important effect on the forming of myelofibrosis. Consequently, ROS plays a significant role in the fibrogenesis of tissues and organs. In this review, the relevance between ROS and common tissues and organs fibrosis is summarized.
Animals ; Bone Marrow ; pathology ; Bone Marrow Diseases ; metabolism ; pathology ; Fibrosis ; Humans ; Liver ; pathology ; Liver Cirrhosis ; metabolism ; pathology ; Lung ; pathology ; Pulmonary Fibrosis ; metabolism ; pathology ; Reactive Oxygen Species

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

The theoretical basis of the alcohol precipitation process control was provided, the alcohol precipitation was optimized and the relationship equation was got. The monod glycoside, loganin and paeoniflorin were used as the evaluation indexes to determine the impact factors of alcohol precipitation techniques of Liuwei Dihuang decoction by the Plackett-Burman experimental design and the levels of non-significant factors were identified. Then, Box-Behnken response surface methodology was used to research and discuss the critical process parameters influence the effect of alcohol precipitation and draw interaction between key process parameters and the correlation equation with index components. Through the establishment and solving the quadratic regression model of composite score, the optimum preparation conditions of alcohol precipitation techniques of liuwei were as follows: stirring speed was 580 r x min(-1), standing time was 17 hours, alcohol concentration was 34%, the density of Liuwei Dihuang decoction was 1.13. The response surface methodology for optimized alcohol precipitation techniques of Liuwei Dihuang decoction is. reasonable and feasible.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry

Objective:To explore the clinical efficacy of sinew-regulating and bone-setting manipulation combined with functional exercise to treat mild to moderate rotator cuff injury (RCI).Methods:Forty patients with mild to moderate RCI were randomly divided into a treatment group and a control group,with 20 cases in each group.The treatment group was given sinew-regulating and bone-setting manipulation combined with functional exercise,and the control group was given the same manipulation as the treatment group.The therapeutic effect on the functional improvement was assessed after treatment according to pain threshold,shoulder range of motion (ROM) including flexion,abduction,internal rotation and external rotation,and the University of California at Los Angeles (UCLA) shoulder rating score.Results:Compared with the same group before treatment,the pain threshold,shoulder flexion,abduction,internal and external rotations increased after the treatment in the two groups (all P＜0.05).The UCLA shoulder rating score increased (both P＜0.05).The improvement in the treatment group is significantly better than that in the control group with a statistical significance (P＜0.05).The total effective rate was 95.0％ in the treatment group,was higher than 65.0％ in the control group (P＜0.05).Conclusion:The sinew-regulating and bone-setting manipulation combined with functional exercise relieve the pain of patients with mild to moderate RCI,increase the motion of shoulder joints,and improve the quality of life of patients.The curative effect is better than the treatment of simple sinew-regulating and bone-setting manipulation.

OBJECTIVEIn aid of MR imaging diagnosis of ankylosing spondylitis (AS), MRI findings of sacroiliac (SI) joints in volunteers were compared with those in patients with AS, with special reference to the fat findings.

METHODS18 normal volunteers and 52 AS patients underwent the MRI examinations with the same protocols, i.e., SE T1WI, FSE T2WI, GE T2*WI sequences were taken using a 0.3T imager. All the images were observed and analyzed by two radiologists.

RESULTSSymmetric or patchy distribution fat component either in sacrum and/or ilium was detected in 52.8% of SI joints of volunteers. Thin zone distribution of fat was seen in 35.6% of SI joints of AS patients, which was adjacent to the erosion and sclerosis. In another 25.0% of SI joints of AS patients, the distribution of fat component was found similar to that of the normal volunteers. Cartilage changes were detected in 85.6% of the SI joints of AS patients. In addition, bone erosion, sclerosis and subchondral bone marrow edema were also detected.

CONCLUSIONSubchondral fat infiltration associated with erosion and sclerosis should be considered as sacroiliitis. The knowledge of the characteristics of fat component could be helpful for evaluating sacroiliac joints.

Adipose Tissue ; pathology ; Adolescent ; Adult ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Sacroiliac Joint ; pathology ; Spondylitis, Ankylosing ; diagnosis ; pathology

OBJECTIVETo compare MR imaging with radiographs in detection of the erosions of the wrists in patients with rheumatoid arthritis (RA).

METHODSIn 50 wrists of 25 patients with RA, radiographs of AP projection and MR imaging examination were performed within 1 week. The MRI sequences were SE T1-weighted image, FSE T2-weighted image, and GE T2* weighted image in coronal plane. Gd-DTPA enhanced SE T1-weighted image with same parameters of pre-contrast administration SE T1WI was performed in 25 wrists, which showed prominent lesions in each 25 patients.

RESULTSIn 50 wrists of 25 patients with RA, bone erosions were found by MR images compared to radiographs in 46 versus 14 wrists, respectively. There were 170 sites of carpal erosions found by MR images, while radiographs found only 23 sites of carpal erosions. Among them triquetrum and lunate were the most frequently involved sites. Significant difference (P < 0.001) was statistically found between conventional radiography and MR imaging in detection of erosions of the wrists of RA. MR imaging also permitted demonstrating synovitis and joint effusion in the wrists. Gd-DTPA-enhanced images allowed distinguishing pannus changes from joint effusion.

CONCLUSIONMRI is superior to conventional radiography with respect to detection of bone erosion in wrists. For those whose conventional radiographs show suspected RA, MRI examination should be advised for the early diagnosis.

Adult ; Aged ; Arthritis, Rheumatoid ; diagnosis ; diagnostic imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Wrist Joint ; pathology

To determine the optimum process conditions for dry granulating technique of Qibai Pingfei granule, granule excipient type, rolling wheel speed and pressure and feeding speed were studied. Taking shaping rate at a time, moisture absorption and dissolubility as index, the type and amount of granule excipient were determined. In addition, taking shaping rate at a time as index, parameters of rolling wheel speed and pressure and feeding speed were researched through single factor test and response surface methodology. The optimum parameters were as follows: lactose as excipient, dry extract powder to excipient at 1:2, rolling wheel speed and pressure at 10.9 Hz and 6.4 MPa and feeding speed at 7.2 Hz. After validation of three batches pilot-scale production, the optimum processing parameters for dry granulating technique of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
Drugs, Chinese Herbal ; Powders ; Technology, Pharmaceutical ; methods

OBJECTIVETo detect the expression of RANKL and OPG protein in the giant cell lesions of jaw and to study the mechanism of this lesion.

METHODSRANKL and OPG were detected by immunohistochemistry (SP) in 24 paraffin-embedded and 2 frozen specimens of central giant cell lesion of jaw.

RESULTSRANKL signals were strongly positive in the vascular epithelial cells. They also could be found in fibrous stroma, bone matrix, and stromal spindle cells, even in some cytomembrane of multinucleated giant cells. OPG was detected in multinucleated giant cells and a fraction of round mononuclear cells.

CONCLUSIONSActive vascular epithelial cells are contributed to the formation of multinucleated giant cells through regulating RANKL, and RANKL could play its role by paracrine and autocrine, which might be inhibited by OPG.

Giant Cells ; metabolism ; pathology ; Humans ; Jaw Diseases ; metabolism ; pathology ; Osteoclasts ; metabolism ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism

Objective To investigate the potential of adult mesenchymal stem cells (MSCs) derived from human bone marrow to undergo cardiomyogenic differentiation after exposure to 5-azacytidine (5-aza) in vitro. Methods A small bone marrow aspirate was taken from the iliac crest of human volunteers, and hMSCs were isolated by 1.073g/mL Percoll and propagated in the right cell culturing medium as previously described. The phenotypes of hMSCs were characterized with the use of flow cytometry. The hMSCs were cultured in cell culture medium (as control) and medium mixed with 5-aza for cellular differentiation. We examined by immunohistochemistry at 21 days the inducement of desmin, cardiac-specific cardiac troponin I (cTnI), GATA 4 and connexin-43 respectively. Results The hMSCs are fibroblast-like morphology and express CD44+ CD29+ CD90+ / CD34- CD45- CD31- CD11a. After 5-aza treatment, 20-30% hMSCs connected with adjoining cells and coalesced into myotube structures after 14days. Twenty-one days after 5-aza treatment, immunofluorescence showed that some cells expressed desmin,GATA4, cTnI and connexin-43 in 5,10 μmol/L 5-aza groups, but no cardiac specific protein was found in neither 3μmol/L 5-aza group nor in the control group. The ratio of cTnI positively stained cells in 10 μmol/L group was higher than that in 5 μmol/L group (65.3 ± 4.7% vs 48.2 ± 5.4%, P ＜ 0.05). Electron microscopy revealed that myofilaments were formed. The induced cells expressed cardiac-myosin heavy chain (MyHC) gene by reverse transcription-polymerase chain reaction (RT-PCR). Conclusions Theses findings suggest that hMSCs from adult bone marrow can be differentiated into cardiac-like muscle cells with 5-aza inducement in vitro and the differentiation is in line with the 5-aza concentration. (J Geriatr Cardiol 2004;1(2) :101-107. )