Synaptosomes were isolated from pig brain by homogenization, differential centrifugation and sucrose gradient centrifugation. After synaptosome lysis in hypoosmotic buffer, the plasma membrane vesicles were collected. Following the solubilization of plasma membrane vesicles in Triton X-100, the solubilized protein was applied to calmodulin affinity chromatography colurnn, and the delipidated plasma membrane Ca2 + -ATPase was purified to nearly homogeneity. The novel feature of this purification is the use of large affinity column and heavy washing to facilitate the purified Ca2+ -ATPase with higher activity and protein yield. The specific activity of the purified Ca2+ -ATPase was recovered to a maximum of 3.32 μmol· mg-1· min-1 after incubation with asolectin. Silver staining of SDS-PAGE revealed a single protein band around Mr 140 000, showing the purity was over 90 %. Different Ca2 + concentrations dramatically affect the specific activity of Ca2 + -ATPase.

The Na+/H+ antiporter in vacuolar membranes transports Na+ from the cytoplasm to vacuoles using a pH gradient generated by proton pumps, which could reduce Na+ toxicity. It is uncertain that whether the woody plants have the same mechanism. Through differential centrifugation and sucrose density gradient centrifugation, tonoplast vesicles were isolated from Populus tremula calli broken by blender. After establishing pH gradient by V-ATPase, Na+ could dissipate the pH gradient, which indicates that there is Na+/H+ antiporter in the tonoplast vesicles from Populus tremula calli (Km=11.4 mmol/L). Amiloride could inhibit the Na+/H+ antiporter activity. The antiporter could transport Na+ and K+, the affinity for Na+ is higher. Salt stress decreased Km and Vmax.

Objective To appraise facial skin's quality conveniently and correctly.Methods The quality appraisal model for facial skin image was established based on improved BP nerve network algorithm,and the model was trained with the intrinsic pattern of the training sample,then the appraisal result was output after inputting indexes on the skin.Results For 23 sets of samples,the appraisal accuracy reached 95%.Conclusion The network model,with a high accuracy and a low error rate,is applicable to facial skin image quality appraisal.

It is generally believed that sphingomyelin- and cholesterol-enriched microdomains can be isolated as detergent-resistant membranes (DRMs) from plasma membrane and organelle membranes. Here the isolation of microdomains from porcine cardiac sarcoplasmic reticulum (SR) membranes is described. These SR-derived detergent-resistant membranes (SR-DRMs) enriched in sphingomyelin and cholesterol have a low buoyant density. Both ganglioside GM1 and caveolin-3 in the SR, known as a marker protein of caveolae at plasma membrane are present in the SR-DRMs. It was demonstrated that significant amount of SERCAs together with caveolin-3 associates with SR-DRMs, and are fully functional. The results suggested that SR membranes like other biomembranes,such as the plasma membrane, contained microdomains.

Four methods were compared to purify Fo from porcine h ear t mitochondria. The best results were obtained by the following method: after re moving F1-ATPase with NaBr incubation from submitochondrial FoF1-ATPase, Fo was solubilized with CHAPS and purified by sucrose density gradient centri fugation. SDS-PAGE with silver staining showed about 85% purity of the isolated Fo and 9 different subunits including b, OSCP, d, a, e, F6, IF1, A6L and c. The purified Fo was then incorporated into asolectin liposomes, the reconst ituted Fo showed higher H+ translocation activity and after Fo was reconst ituted with F1-ATPase, the resulted FoF1-ATPase complex exhibited high A TP hydrolysis activity and high sensitivity to oligomycin. The results provide e vidence for successful purification, reconstitution of Fo with high H+ trans location activity and its relationship with phospholipids.

3D reconstruction from medical images is widely used in diagnose, radiotherapy surgery programming and medical research. This paper deals with the framework and key techniques of 3D reconstruction system of medical images. Methods of automatic and manual segmentation of medical images are designed. Data structure of the system is proposed. The system data are encapsulated in object-oriented paradigm and the hierarchical structure of classes is presented for reconstructing 3D geometry models. The 3D surface geometry model is constructed by using MT algorithm from the tissues extracted by automatic or manual segmentation. An edge collapse algorithm of mesh simplification is proposed and implemented, and the surface model constructed by MT algorithm is simplified by using this algorithm, After reduction of 90 percent triangles, the simplified model still maintains the original feature. It can speed up surface rendering of the models.
Algorithms ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional

OBJECTIVETo investigate and compare the efficacy and safety of two kinds of Qinbai ointment prepared conventionally and finely (C-QBO and F-QBO) in treating psoriasis in the active stage of blood-heat syndrome type, and to observe their influences on vaginal epithelial cell mitosis, also on keratinization of caudal flakes in mice.

METHODS

CLINICAL OBSERVATIONAdopting randomized, single-blinded, controlled design, 93 patients administered orally with Liangxue Huoxue Decoction were randomized into three groups treated externally with C-QBO, F-QBO and white vaseline respectively, applied twice a day for 8 weeks. The safety was checked and the changes of modified psoriasis area severity index (PASI), as well as the conditions of skin lesions (size, erythema, infiltration, squama, and itching) and symptom improving time were compared. Experimental study 1: Mice were randomized into the model group, the C-QBO group, the F-QBO group and the blank group, 7 in each group. Their vaginal epithelial cell mitosis indices were compared after 3 days of treatment with corresponding remedies. Experimental study 2: Mice were randomized into the C-QBO group, the F-QBO group and the blank group, 7 in each group. After the mice had been treated with corresponding remedies for 28 days, the granular layer formation in their caudal scales was compared.

RESULTS

CLINICAL OBSERVATIONThe markedly effective rate was 63.3% (25/30) in the C-QBO group, 66.7% (20/30) in the F-QBO group, and 36.7% (11/30) in the control group. No statistical difference was showen between the two QBO treated groups (P > 0.05), but that in both of them was significantly different from that in the control group (P < 0.05). PASI scores lowered after treatment in all the three groups (P < 0.01), but the improvement in the two QBO groups was better than that in the control group (P < 0.05); the conditions of skin lesion were improved in all groups, but the improvements were more significant in the two QBO groups in terms of squama, infiltration and itching (P < 0.05), in aspects of improving time on erythema, infiltration and itching, especially the itching, F-QBO was superior to the C-QBO. Experimental study 1: The mitosis index in both QBO groups was lower than that in the blank group and the model group (P < 0.01). Experimental study 2: Number of scales with granular layer formation was higher in the two QBO groups than in the blank group (P < 0.01).

CONCLUSIONC-QBO and F-QBO can effectively relieve the skin lesion of psoriasis patients in the active stage of blood-heat type, and they could also promote the formation of epithelial granular layer in the caudal scales of mice.

Adult ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred Strains ; Middle Aged ; Ointments ; Phytotherapy ; Psoriasis ; diagnosis ; drug therapy ; Treatment Outcome

BACKGROUND:Resveratrol is a natural antioxidant extracted from plants.Its mechanism of improving exercise-induced fatigue mainly focuses on the protective effect against oxidative stress and inflammation.In this study,the protective mechanism of resveratrol on exercise-induced fatigue was mainly discussed from the perspective of gluconeogenesis. OBJECTIVE:To investigate the effect of resveratrol on gluconeogenesis in exercise-induced fatigue rats. METHODS:After 1 week of adaptive training,male Sprague-Dawley rats were randomly divided into 4 groups with 12 rats in each group:blank control group,resveratrol group,exercise group,resveratrol + exercise group.Weight-bearing swimming training was used to simulate long-term medium-high intensity exercise.After swimming with a weight of 5%for 1 hour every day,50 mg/kg resveratrol solution or the same volume of dimethyl sulfoxide solvent were given orally,6 days a week,for a total of 6 weeks.Samples were collected 24 hours after the last exercise,and the levels of urea nitrogen,creatine kinase,blood glucose,liver glycogen and lactic acid and pyruvate in liver tissue were detected by the kit.The activity of phosphoenolpyruvate carboxykinase was detected by microassay,and the activity of glucose-6-phosphatase was detected by enzyme-linked immunosorbent assay.Real-time fluorescence quantitative PCR was used to detect the gene expression of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α. RESULTS AND CONCLUSION:In the exercise group,plasma urea nitrogen and creatine kinase levels of rats were significantly increased(both P<0.05),liver lactate and pyruvate levels and lactate/pyruvate ratio were significantly increased(all P<0.01),and blood glucose and liver glycogen contents were significantly decreased(both P<0.01).Resveratrol supplementation could effectively improve the above conditions.Exercise significantly decreased the activities of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase(P<0.01,P<0.05),and resveratrol supplementation significantly increased the activity of phosphoenolpyruvate carboxykinase in liver tissue(P<0.01).The mRNA expression levels of silent information regulator 1,cAMP-response element binding protein and peroxisome proliferator-activated receptor-γ coactivator-1α in liver tissue of the exercise group were significantly decreased(all P<0.01),while resveratrol supplementation could significantly increase the gene expression levels of this pathway.To conclude,resveratrol can relieve exercise-induced fatigue caused by long-term medium-high intensity exercise,and its mechanism may be related to up-regulating the gluconeogenesis regulatory pathway,improving rate-limiting enzyme activity,promoting liver gluconeogenesis,and increasing blood glucose and liver glycogen levels.

Macroautophagy (referred to as autophagy hereafter) is a major intracellular lysosomal degradation pathway that is responsible for the degradation of misfolded/damaged proteins and organelles. Previous studies showed that autophagy protects against acetaminophen (APAP)-induced injury (AILI) via selective removal of damaged mitochondria and APAP protein adducts. The lysosome is a critical organelle sitting at the end stage of autophagy for autophagic degradation via fusion with autophagosomes. In the present study, we showed that transcription factor EB (TFEB), a master transcription factor for lysosomal biogenesis, was impaired by APAP resulting in decreased lysosomal biogenesis in mouse livers. Genetic loss-of and gain-of function of hepatic TFEB exacerbated or protected against AILI, respectively. Mechanistically, overexpression of TFEB increased clearance of APAP protein adducts and mitochondria biogenesis as well as SQSTM1/p62-dependent non-canonical nuclear factor erythroid 2-related factor 2 (NRF2) activation to protect against AILI. We also performed an unbiased cell-based imaging high-throughput chemical screening on TFEB and identified a group of TFEB agonists. Among these agonists, salinomycin, an anticoccidial and antibacterial agent, activated TFEB and protected against AILI in mice. In conclusion, genetic and pharmacological activating TFEB may be a promising approach for protecting against AILI.