This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

This study was to investigate the permeability and absorbability of capsaicin cubosome across abdominal skin of the SD rats in vitro. Diffusion of capsaicin cubosome and cream was performed with the modified Franz diffusion cell technique. The capsaicin cubosome showed no enhancement of skin permeation within 24 hours. However, the deposition amounts of capsaicin in the rat skin in the cubosome group was markedly higher than those in the commercial cream group (P < 0.01). Cubosome showed excellent characetristic of skin-targed which could be a good carrier for the local transdermal drug delivery system.
Administration, Cutaneous ; Animals ; Capsaicin ; administration & dosage ; chemistry ; Kinetics ; Male ; Particle Size ; Permeability ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; metabolism ; Skin Absorption

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

Objective:To investigate the mechanism of liver and lung injury in mouse septic models.Methods:Twenty-four male Kunming mice were subjected to cecal ligation and puncture(CLP)or sham operation.The permeability of microvasculature,water contents,activities of myeloperoxidase(MPO)and the apoptosis of microvascular endothelial cells in lung microvasculature and liver sinus were examined 3 h and 12 h after operation.Results:Both the liver and lung showed a significant increase in microvessel permeability at 12 h in CLP group compared with sham operation group.MPO activity and water content in CLP group were obviously higher than those in the sham operation group.The apoptosis of lung microvascular endothelial cells at 12 h in CLP group(5.03?0.92)% was significantly higher than that of control group(3.48?1.21)%(P

AlM:To analyze and compare the effect of femtosecond laser micro - incision corneal stromal lens excision ( SMlLE) and excimer laser in situ keratomileusis ( LASlK) in the treatment of myopia after operation, to explore the safety, operability and prediction of SMlLE.METHODS:ln this prospective clinical controlled study, 100 cases ( 200 eyes ) received SMlLE and 100 cases ( 200 eyes) undergone LASl in our hospital in the same period were selected. Uncorrected visual acuity, diopter, corrected visual acuity, slit lamp examination, intraocular pressure and corneal anterior segment OCT, corneal topography (Obscan ll) of two groups in 1d, 1wk, 1, 3, 6mo, 1a were compared. lndependent samples t test was used for data analysis.RESULTS:1) Postoperative slit lamp examination:after 1d in SMlLE group, there were less eyes had corneal layer between mild cloudy or edema; postoperative 1wk corneal layer disappeared, cornea became clear and transparent. 2 ) Postoperative vision recovery: 1d after operation, vision recovery in LASlK group was better than that in SMlLE group, the difference was statistically significant (P<0. 01), there were no significant differences at 1wk, 1, 3, 6mo, 1a after operation ( P>0. 05 ). 3 ) Obscan ll examination: graphics in the SMlLE group was more regular and placed in the center, no eccentric and irregular graphics, better than that in the LASlK group. 4) Anterior segment OCT examination:postoperative corneal flap in the SMlLE group was more uniform and accurate, but it was thin in the center and slightly thick the peripheral part in the LASlK groups. 5 ) Postoperative visual quality assessment used subjective questionnaire survey. The two groups had statistically significant difference on 4 points and 1 points (P<0. 05). Complains in the LASlK groups were more that that in the SMlLE group. While, no complain of the SMlLE group was higher than that of the LASlK group. Glare of postoperative patients with night vision and dark environment in the SMlLE group was better than that of the LASlK group.CONCLUSlON: SMlLE is safe, effective, stable and predictable for the correction of myopia.

The purpose of this study is to investigate the preparation of hydroxycamptothecine (HCPT)-loaded cubic crystal liquid embolic precursor solution, and evaluate its in vitro embolic efficiency. Phytantriol was used as cubic crystal liquid embolic material, and the optimal formulation was selected according to ternary phase diagram. Polarized light microscopy, differential scanning calorimetry, and small angle X-ray scattering (SAXS) were used to characterize the cubic crystal structure. High performance liquid chromatography and X-ray diffraction analysis were used to investigate the lactone ring of HCPT. In vitro dissolution was preliminary evaluated, and the simulation embolic model was constructed to evaluate the embolic efficiency of precursor solution. Meanwhile, the gelation time and adhesion force were investigated. The results showed that HCPT-loaded precursor solution for embolization had been successfully prepared with low viscosity which was injectable. The precursor solution could transform into Pn3m structure liquid crystal phase gel rapidly when contracting with excess water. The formed HPCT gel remained its lactone form as the same in precursor solution, and expressed the good ability to block the saline flow, and HCPT could keep sustained releasing drug over 30 days. The prepared drug-loaded embolic precursor solution showed a promising potential for vascular embolization and application in clinical treatment of tumor.
Antineoplastic Agents, Phytogenic ; chemistry ; Calorimetry, Differential Scanning ; Camptothecin ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; chemistry ; Fatty Alcohols ; chemistry ; Liquid Crystals ; Scattering, Small Angle ; Water ; X-Ray Diffraction

Hot-melt extrusion was applied to prepare mesoporous silica/ethylcellulose mini-matrix for sustained release, and fenofibrate was used as a model drug, ethylcellulose and xanthan gum were chosen as sustained-release agent and releasing moderator, respectively. This novel matrix obtained the controlled release ability by combining mesoporous silica drug delivery system and hot-melt extrusion technology. And mesoporous silica particle (SBA-15) was chosen as drug carrier to increase the dissolution rate of fenofibrate in this martix. Scanning electron microscope, transmission electron microscope, small angle X-ray powder diffraction and N2 adsorption-desorption were introduced to determine the particle morphology, particle size and pore structure of the synthesized SBA-15. The results showed that SBA-15 had a very high Brunauer-Emmett-Teller specific surface area, a narrow pore size distribution, large pore volume and a ordered two-dimensional hexagonal structure of p6mm symmetry. Differential scanning calorimetry and X-ray powder diffraction results demonstrated that fenofibrate dispersed in an amorphous state inside the pores of the mesoporous silica which contributed to the improvement in the dissolution rate. The drug release of mini-matrices was influenced by ethylcellulose viscosity grades and xanthan gum concentration, which increased with the increasing of xanthan gum concentration and decreasing of ethylcellulose viscosity. Mini-matrix containing 22% xanthan gum exhibited a good sustained release performance, and the drug release behavior followed the first-order kinetics.
Adsorption ; Calorimetry, Differential Scanning ; Cellulose ; analogs & derivatives ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Particle Size ; Porosity ; Powder Diffraction ; Powders ; Silicon Dioxide ; Solubility ; X-Ray Diffraction

BACKGROUNDIodine staining during endoscopy has been successfully used to detect early carcinomatous and precancerous lesions in the esophagus, cervix, and oral cavity. The objective of this study was to determine the diagnostic accuracy of fiberoptic ductoscopy (FDS) plus in vivo iodine staining for intraductal proliferative lesions of the breast.

METHODSWe performed periodic acid-Schiff (PAS) and in vitro iodine staining on 52 and 64 specimens of benign mammary hyperplasia, respectively, and 57 and 53 specimens of ductal carcinoma in situ (DCIS), respectively. Next, FDS was performed on 177 recurrent nipple discharge patients who were randomly divided into two groups. One group was iodine-staining group in which 92 patients were randomly selected to undergo iodine staining during FDS, and the remaining 85 were assigned to the control group. Biopsy specimens of suspicious lesions were obtained and subjected to histopathological examination.

RESULTSFollowing PAS staining, benign mammary hyperplasia lesions were positively stained, while negligible PAS positivity was observed in the DCIS lesions (P < 0.05). Following in vitro iodine staining, benign mammary hyperplasia specimens appeared dark brown, whereas DCIS samples appeared significantly lighter or unstained. Compared with the pathological examination results, FDS with iodine staining showed an agreement rate in the diagnosis of ductal intraepithelial neoplasia (DIN), sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and Youden index of 97.82%, 98.83%, 83.33%, 5.93, 0.014, and 0.8216, respectively; the corresponding values for FDS without iodine staining were 88.24%, 89.16%, 50.00%, 1.78, 0.217, and 0.3916, respectively.

CONCLUSIONFDS with iodine staining was superior to conventional FDS for the diagnosis of DIN and is valuable for breast cancer prevention.

Adult ; Aged ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma, Ductal, Breast ; diagnosis ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; diagnosis ; pathology ; Female ; Fiber Optic Technology ; Humans ; Hyperplasia ; Iodine ; Middle Aged ; Periodic Acid-Schiff Reaction ; Staining and Labeling

OBJECTIVESTo study the role of cyclooxygenase 2 (COX 2) and prostaglandin I2 (PGI2) in the development of portal hypertensive gastropathy (PHG).

METHODSForty Wistar rats were divided into surgery group (32) and control group (8). Partial portal vein ligation method was used to narrow the sectional area of portal vein to establish the experimental model of PHG in surgery group rats. Then they were divided into four groups (8 rats in each). The free pressure of portal vein was determined at the 1st, 2nd, 3rd, 4th weeks after the operation, and 8 rats were killed to observe the pathological change of gastric mucosa. The levels of 6-keto-PGF1 alpha, a stable metabolite of PGI2, were determined by radioimmunoassay in gastric mucosa homogenate and the blood of portal vein. The expression of COX 2 in gastric mucosa was determined by immunohistochemistry.

RESULTSThe free pressure of portal vein increased rapidly after partial portal vein ligation and maintained a high stable level after 1 week. They were (2.40+/-0.15) kPa, (2.38+/-0.17) kPa, (2.52+/-0.21) kPa, and (2.46+/-0.17) kPa at the 1st, 2nd, 3rd, and 4th weeks after partial portal vein ligation, while it was (0.90+/-0.16) kPa in control group (t>or=17.356, P<0.05). The gastric mucosa appeared pale, edema, hyperaemia, surface erosion, punctate hemorrhage and these lesions were more apparent with the time after the operation. The pathological examination showed that the gastric mucosa and submucosa thickened. The vessels of gastric mucosa and submucosa expanded and increased. There were lymphocytes and neutrophils infiltration around the vessels in the gastric mucosa and submucosa. The 6-keto-PGF1 alpha levels in gastric mucosa and the blood of portal vein increased rapidly and maintained a high level after partial portal vein ligation,which were higher than those in control group (104.52pg/ml+/-25.11pg/ml vs 73.62pg/ml+/- 20.33pg/ml, t=2.710, P<0.05; 180.21pg /ml+/-37.56pg /ml vs 142.11pg /ml+/-31.51pg /ml, t=2.198, P<0.05). The results of immunohistochemistry showed that the intensity and degree of the COX 2 staining in gastric tissue increased at the 1st, 2nd, 3rd, 4th weeks after partial portal vein ligation, while the COX 2 in control group rats was negative.

CONCLUSIONSThe expression of COX 2 and PGI2 in gastric tissue increased in portal hypertension. PGI2 as an inflammatory medium, damages the gastric mucosa by expanding vessels and other mechanisms in portal hypertension. It may be one of the important factors contributing to the development of PHG.

Animals ; Cyclooxygenase 2 ; Disease Models, Animal ; Epoprostenol ; physiology ; Hypertension, Portal ; complications ; pathology ; Isoenzymes ; physiology ; Male ; Prostaglandin-Endoperoxide Synthases ; physiology ; Rats ; Rats, Wistar ; Stomach Diseases ; etiology ; pathology

OBJECTIVETo evaluate the effect of Wulongdan on the learning and memory abilities of rats with chronic cerebral ischemia and explore the mechanisms.

METHODSMale SD Rat models of chronic cerebral ischemia were established by permanent ligation of the bilateral carotid arteries. Three weeks after the operation, the rats were randomly divided into sham-operated group, chronic cerebral ischemia group (model group), high-dose drug group, low-dose drug group and Yinxingye group and received the corresponding treatments on a daily basis for 5 consecutive weeks. Morris water maze was used to assess the learning and memory abilities of the rats, and Western blotting was carried out for detecting the expressions of NR1 and NR2B in the hippocampus and cerebral cortex.

RESULTSCompared with the model group, the rats in high-dose drug, low-dose drug and Yinxingye groups showed significantly shorter time of finding platform in Morris water maze test (P<0.05 or 0.01). The rats in the model group showed significantly lowered expressions of NR1 and NR2B of the cortex and hippocampus than those in the sham-operated group (P<0.01). In comparison with the model group, the high-dose Wulongdan group and Yinxingye group both showed significantly increase expressions of NR1 and NR2B (P<0.01), but their expression levels still remained significantly lower than those in the sham-operated group (P<0.01).

CONCLUSIONWulongdan can enhance the learning and memory abilities of rats with chronic cerebral ischemia, the mechanisms of which may involve increased expressions of NR1 and NR2B in the hippocampus and cortex.

Animals ; Brain Ischemia ; drug therapy ; psychology ; Cerebral Cortex ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; drug effects ; metabolism ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism