Preparation and in vitro evaluation of forsythoside A-loaded exosomes / 中国中药杂志

A drug delivery system of forsythoside A-loaded exosomes(FTA-Exos) with high biocompatibility and low immunogenicity was established to investigate its impact on the migration of human lung epithelial adenocarcinoma A549 cells. The exosomes from A549 cells were extracted and purified by ultra-high speed centrifugation and ultrafiltration. FTA-Exos were prepared by ultrasonic incubation, and characterized by particle size analysis, transmission electron microscopy, and Western blot assay. The uptake of FTA-Exos by A549 cells was observed under the laser confocal microscope, and the impact of FTA-Exos on the migration of A549 cells was investigated by cell scratch assay. The results showed that the average particle size of the prepared FTA-Exos was(138.90±2.37) nm, which increased slightly after drug loading. The PDI was 0.291±0.013, and the average potential was(-10.1±0.66) mV. The FTA-Exos were spheroidal in appearance as observed by transmission electron microscope, with an obvious saucer-like double-layer membrane. Western blot assay indicated that the specific proteins CD63 and Alix were both expressed in exosomes. The laser confocal microscopy suggested that FTA-Exos were taken up by A549 cells and stably maintained in the cell for 4-8 h, and the fluorescence was significantly enhanced at 4 h. The scratch assay showed that the inhibitory effect of FTA-Exos on the migration of A549 cells was significantly stronger than that of forsythoside A(P < 0.05). In conclusion, the drug delivery system of FTA-Exos established in this study had good stability, reliable preparation process, and potent inhibitory effect on the migration of A549 cells in vitro, which can provide an important reference for subsequent in-depth research and application.

Bioinformatics Analysis of Up-regulated Differential Genes in Rats post Spinal Cord Injury / 中国康复理论与实践

Objective:To investigate the differential expression and gene functions of up-regulated genes in rats with spinal cord injury. Methods:Female Sprague-Dawley rats' model of spinal cord injury was established with the modified Allen's method. Gene chip technology was used to detect the variation of differentially expressed genes in the spinal cord after spinal cord injury in rats. The differences in genes, functional localization and pathways were analyzed with gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway database. Results:The results of total RNA quality in spinal cord segment were qualified. Gene chip results showed that there were 1874 differentially up-regulated genes and 2348 differentially down-regulated genes. Bioinformatics was used to analyze differentially up-regulated genes in terms of biological processes, cellular components, and molecular functions. The differentially up-regulated genes were involving apoptosis, immune response, inflammation, etc., pathway analysis mainly showed the differentially up-regulated genes involved phosphoinositide 3-kinase protein kinase B signaling pathway and Toll-like receptor signaling pathways. Conclusion:Differentially up-regulated genes may be involved in secondary reactions following spinal cord injury, such as inflammation, immune response and hypoxia, and then further affect motor function and sensory function.

Effects of propofol on biological characteristics of colorectal cancer cells / 中国病理生理杂志

AIM:To investigate the effect of propofol on the viability,invasion ability and apoptosis of colorec-tal cancer cells.METHODS:Propofol at 10,25,50 and 100 μmol/L was used to treat LoVo cells for 72 h,and propofol at 100 μmol/L was used to treat the LoVo cells for 12,24,48 and 72 h.The cell viability was measured by CCK-8 assay. The invasion ability of the LoVo cells treated with propofol at 100 μmol/L for 72 h was detected by Transwell assay.The cell cycle distribution and cell apoptotic rate were analyzed by flow cytometry.The protein levels of matrix metalloproteinase (MMP)-2,MMP-9,cleaved caspase-3,Notch1 and hairy and enhancer of split 1(Hes1)were determined by Western blot.RESULTS:Propofol inhibited LoVo cell viability.The cell invasion ability,S stage cells,and the protein levels of MMP-2,MMP-9,Notch1 and Hes1 in propofol group were significantly lower than those in control group,and the apoptotic rate,G0/G1cells and the protein level of cleaved caspase-3 were significantly higher than those in control group(P<0.01).CONCLUSION:Propofol inhibits the viability and invasion ability of colorectal cancer LoVo cells,blocks cell cy-cle and induces apoptosis.The mechanism is related to down-regulation of Notch1 signaling pathway.

The disinfection efficiency observation of a compound disinfectant spray on dental impression and plaster / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The disinfection efficiency of a compound disinfectant spray with trichioro hydroxyl diphenyl ether on dental impression and plaster model, which have been contaminated by pathogens, were evaluated in this study.</p><p><b>METHODS</b>As experimental group, germ-free alginate impressions and plaster models were sprayed with the compound disinfectant of different density trichloro hydroxyl diphenyl ether or indophors for 5, 10 and is mm, after which were smeared with five tested pathogens, including Staphylococcus acre us, Escherichia cali, Saccharomyces albicans, Streptococcus mutans and black spore variants of Bacillus subtilis. The colonies were counted after sampling, inoculate and culture, which were used to deduce the killing logarithm value as the standard of the disinfecting efficiency.</p><p><b>RESULTS</b>the compound disinfectant spray with 3000 mg x L(-1) triebloro hydroxyl diphenyl ether was effective to all tested pathogens for 10 mm whatever on the impressions or the plaster models. The disinfectant spray with tame concentration was more effective on the alginate impression than on the plaster model in the same time (P = 0.000).</p><p><b>CONCLUSION</b>The compound disinfectant spray with trichioro hydroxyl diphenyl ether is an effective antiseptics for alginate impressions and plaster models.</p>

Combined slower plasma exchange and continuous veno-venous hemofiltration with a parallel circuit in the treatment of chronic severe viral hepatitis B patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the clinical effects of combined slower plasma exchange (PE) and continuous veno-venous hemofiltration (CVVH) with a parallel circuit in the treatment of chronic severe viral hepatitis B patients.</p><p><b>METHODS</b>104 patients with chronic severe viral hepatitis B were divided into three groups: 44 patients were treated with a parallel circuit of combined slower plasma exchange and continuous veno-venous hemofiltration (group A), 30 patients were treated with plasma exchange (group B), and 30 patients received routine treatment (group C). Efficacy of treatment and survival rate in three groups were investigated. The levels of cytokine, plasma sodium concentration and pH value were examined before and after artificial liver support system treatment.</p><p><b>RESULTS</b>In group A, 7 of 9 patients in coma regained normal consciousness, 6 of 9 patients with hepatorenal syndrome restored renal function, hyponatremia was improved, the balance of pH value was corrected, tumor necrosis factor (TNF)-alpha level was decreased, and the total survival rate was 56.82%. In group B, 2 of 7 patients in coma regained normal consciousness, 1 of 5 patients with hepatorenal syndrome restored renal function. Hyponatremia, pH value and TNF-alpha level were not changed; the total survival rate was 33.33%. Both IL-1 and IL-6 levels were significantly decreased after treatment in group A. IL-10 level was increased in both group A and group B. In group C, 1 of 6 patients regained normal consciousness from coma, none of them restored renal function, and the total survival rate was 16.67%.</p><p><b>CONCLUSIONS</b>Combined slower PE and CVVH with a parallel circuit is a new, safe and effective non-biological artificial liver in the treatment for chronic severe viral hepatitis B patients.</p>

Determination of soluble solid content and acidity of loquats based on FT-NIR spectroscopy / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The near infrared (NIR) spectroscopy technique has been applied in many fields because of its advantages of simple preparation, fast response, and non-destructiveness. We investigated the potential of NIR spectroscopy in diffuse reflectance mode for determining the soluble solid content (SSC) and acidity (pH) of intact loquats. Two cultivars of loquats (Dahongpao and Jiajiaozhong) harvested from two orchards (Tangxi and Chun'an, Zhejiang, China) were used for the measurement of NIR spectra between 800 and 2500 nm. A total of 400 loquats (100 samples of each cultivar from each orchard) were used in this study. Relationships between NIR spectra and SSC and acidity of loquats were evaluated using partial least square (PLS) method. Spectra preprocessing options included the first and second derivatives, multiple scatter correction (MSC), and the standard normal variate (SNV). Three separate spectral windows identified as full NIR (800approximately2500 nm), short NIR (800approximately1100 nm), and long NIR (1100approximately2500 nm) were studied in factorial combination with the preprocessing options. The models gave relatively good predictions of the SSC of loquats, with root mean square error of prediction (RMSEP) values of 1.21, 1.00, 0.965, and 1.16 degrees Brix for Tangxi-Dahongpao, Tangxi-Jiajiaozhong, Chun'an-Dahongpao, and Chun'an-Jiajiaozhong, respectively. The acidity prediction was not satisfactory, with the RMSEP of 0.382, 0.194, 0.388, and 0.361 for the above four loquats, respectively. The results indicate that NIR diffuse reflectance spectroscopy can be used to predict the SSC and acidity of loquat fruit.

Measurement of soluble solids content in watermelon by Vis/NIR diffuse transmittance technique / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Watermelon is a popular fruit in the world with soluble solids content (SSC) being one of the major characteristics used for assessing its quality. This study was aimed at obtaining a method for nondestructive SSC detection of watermelons by means of visible/near infrared (Vis/NIR) diffuse transmittance technique. Vis/NIR transmittance spectra of intact watermelons were acquired using a low-cost commercially available spectrometer operating over the range 350~1000 nm. Spectra data were analyzed by two multivariate calibration techniques: partial least squares (PLS) and principal component regression (PCR) methods. Two experiments were designed for two varieties of watermelons [Qilin (QL), Zaochunhongyu (ZC)], which have different skin thickness range and shape dimensions. The influences of different data preprocessing and spectra treatments were also investigated. Performance of different models was assessed in terms of root mean square errors of calibration (RMSEC), root mean square errors of prediction (RMSEP) and correlation coefficient (r) between the predicted and measured parameter values. Results showed that spectra data preprocessing influenced the performance of the calibration models. The first derivative spectra showed the best results with high correlation coefficient of determination [r=0.918 (QL); r=0.954 (ZC)], low RMSEP [0.65 degrees Brix (QL); 0.58 degrees Brix (ZC)], low RMSEC [0.48 degrees Brix (QL); 0.34 degrees Brix (ZC)] and small difference between the RMSEP and the RMSEC by PLS method. The nondestructive Vis/NIR measurements provided good estimates of SSC index of watermelon, and the predicted values were highly correlated with destructively measured values for SSC. The models based on smoothing spectra (Savitzky-Golay filter smoothing method) did not enhance the performance of calibration models obviously. The results indicated the feasibility of Vis/NIR diffuse transmittance spectral analysis for predicting watermelon SSC in a nondestructive way.

Application Fourier transform near infrared spectrometer in rapid estimation of soluble solids content of intact citrus fruits / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Nondestructive method of measuring soluble solids content (SSC) of citrus fruits was developed using Fourier transform near infrared reflectance (FT-NIR) measurements collected through optics fiber. The models describing the relationship between SSC and the NIR spectra of citrus fruits were developed and evaluated. Different spectra correction algorithms (standard normal variate (SNV), multiplicative signal correction (MSC)) were used in this study. The relationship between laboratory SSC and FT-NIR spectra of citrus fruits was analyzed via principle component regression (PCR) and partial least squares (PLS) regression method. Models based on the different spectral ranges were compared in this research. The first derivative and second derivative were applied to all spectra to reduce the effects of sample size, light scattering, instrument noise, etc. Different baseline correction methods were applied to improve the spectral data quality. Among them the second derivative method after baseline correction produced best noise removing capability and yielded optimal calibration models. A total of 170 NIR spectra were acquired; 135 NIR spectra were used to develop the calibration model; the remaining spectra were used to validate the model. The developed PLS model describing the relationship between SSC and NIR reflectance spectra could predict SSC of 35 samples with correlation coefficient of 0.995 and RMSEP of 0.79 degrees Brix.

Combined non-biological artificial liver in treatment of late stage chronic severe hepatitis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the clinical effects of combined non-biological artificial liver in the treatment of late stage chronic severe hepatitis and especially to observe their effects on hepatic encephalopathy, hepatorenal syndrome and disturbance of electrolytes.</p><p><b>METHODS</b>103 chronic severe hepatitis patients were treated with the same medical measures, including plasma exchange. Among them, 63 patients were also treated with combinations of non-biological artificial liver (treatment group), and the other 40 patients served as controls (control group). The efficacy of the treatments and survival rates of the two groups were compared.</p><p><b>RESULTS</b>In the treatment group, the rate of regaining normal consciousness was 72.7%, the rate of electrolyte disorder being rectified was 89.5%, the rate of restoring renal function was 66.7% and the total survival rate was 47.6%. In comparison, in the control group the rate of regaining normal consciousness was 16.7%, the rate of electrolyte disorder being rectified was 42.3%, none of their renal functions were restored and the total survival rate was 22.5%. The differences between the two groups were significant (chi2=6.56, P less than 0.05).</p><p><b>CONCLUSION</b>With other medical treatment, combined non-biological artificial liver can improve the survival rate of severe hepatitis patients.</p>

Integration between M3 muscarinic acetylcholine receptor and connexin 43 as antiarrhythmic targets in rat ventricular myocardium / 药学学报

<p><b>AIM</b>To optimize the method of investigating structural integration between proteins and study the integration between arrhythmia related proteins in molecular level.</p><p><b>METHODS</b>Immunostaining the normal ventricular myocytes was used to observe the distribution of connexin 43 and muscarinic acetylcholine receptor (mAChR). The five mAChR subtypes were precipitated using immunoprecipitation. Then, SDS-PAGE and Western blotting with the anti-connexin 43 antibody were performed to observe whether they were structurally integrated. Further, different concentrations of detergent were used to observe whether this relationship could be broken.</p><p><b>RESULTS</b>The five subtypes of mAChR existed in the cardiac myocyte of the rat, and all the five mAChR subtypes combined with connexin 43. In the normal rat ventricular myocyte membrane, connexin 43 and M3 receptor are co-located. When adding certain concentration of detergent to the membrane protein, the integration between M3 receptor and connexin 43 was broken, and the phosphorylated form of connexin 43 integrated with M3 receptor.</p><p><b>CONCLUSION</b>The results indicated that the structural integration between mAChR and phosphorylation of connexin 43 existed in rat ventricular myocardium, and this integration could be broken by certain concentration of detergent.</p>

Effects of lower fluence pulsed dye laser irradiation on production of collagen and the mRNA expression of collagen relative gene in cultured fibroblasts in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Lower fluence of 585-nm flashlamp-pumped pulsed dye laser has been successfully used as a nonablative technique in the treatment of wrinkles. The objective of this study was to evaluate the effect of the pulsed dye laser (585 nm) on the production of collagen and the mRNA expression of collagen related gene in fibroblasts in vitro.</p><p><b>METHODS</b>Cultured fibroblasts were treated with a 585-nm flashlamp-pumped pulsed dye laser (fluence 3 J/cm(2), 4 J/cm(2), spot size 7 mm, pulse duration 450 micros). The production of collagen and the mRNA expression of transforming growth factor (TGF)-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen alpha1, alpha2 in fibroblasts were investigated by colorimetry or real time polymerase chain reaction.</p><p><b>RESULTS</b>The production of collagen was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm(2) (P < 0.001). The mRNA expression of TGF-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and procollagen I was significantly up-regulated after treatment with a 585-nm flashlamp-pumped pulsed dye laser with a fluence of 3 J/cm(2) (P < 0.001). No significant difference of mRNA expression of SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen was found between controls and fibroblasts treated with pulsed dye laser with a fluence of 4 J/cm(2) (P > 0.05).</p><p><b>CONCLUSIONS</b>Lower fluence (3 J/cm(2)) pulsed dye laser increased the collagen production in fibroblasts by up-regulating TGF-beta1, SMAD2, SMAD3, SMAD4, SMAD7 and type I procollagen mRNA expression. These may be the reason it can be effectively used in the treatment of wrinkles.</p>

Clinical research of plasma exchange with continuous veno-venous hemofiltration in treating mid- and late-stage chronic severe viral hepatitis B patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy and study the mechanism of combining plasma exchange and continuous veno-venous hemofiltration in treating patients with chronic severe viral hepatitis B in their mid- and late stages.</p><p><b>METHODS</b>94 patients suffering from chronic severe viral hepatitis B were divided into three groups. 29 patients were treated with plasma exchange plus continuous veno-venous hemofiltration (group A). 31 patients were treated with plasma exchange alone (group B). 34 patients received routine treatment (group C). The efficacy of treatment and survival rate of the three groups was investigated. Before and after artificial liver support system treatment the levels of cytokine were examined.</p><p><b>RESULTS</b>In group A, hyponatremia improved, the levels of interleukin 8 (IL-8) obviously decreased, the level of IL-10 increased, 5 of the 10 patients in coma regained normal consciousness (50.0%) and their survival rate was 48.3%. In group B, hyponatremia did not change, the level of IL-8 and IL-10 did not change. 2 of 11 patients in coma regained normal consciousness (18.2%) while survival rate was 22.6%. In group C, 1 of 11 patients in coma regained normal consciousness (9.1%) while survival rate was 20.6%.</p><p><b>CONCLUSIONS</b>It shows that plasma exchange with continuous veno-venous hemofiltration in treating patients with mid- and late stage chronic severe viral hepatitis B can increase the survival rate. IL-8 can be significantly removed, IL-10 significantly increased. This combined therapy is easy to practice, and should be used as an artificial liver support system.</p>

Emodin stimulates glucose uptake by HepG2 hepatocyte through activation of PPAR? / 中华内分泌代谢杂志

Objective To construct PPAR?and PPAR?response element (PPRE)-controlled luciferase expression vectors,and to determine whether the traditional Chinese medicine emodin activates PPAR?and improves the glucose uptake by HepG2 hepatocytes.Methods (1) PPAR?and PPRE luciferase expression vectors were constructed and were applied to screen more than 20 ingredients of the traditional Chinese medicine. (2) HepG2 cells were incubated with emodin which can activate PPAR?and PPRE luciferase activity,and the PPAR?mRNA expression level was evaluated by RT-PCR/Southern blot.(3) PPAR?and glucose transporter 2 (Glut2) proteins were determined by Western blot analysis in HepG2 cells treated with emodin.(4) The glucose uptake rate was measured using 2-deoxy-[~3H]-D-glucose in HepG2 cells after treatment with emodin.Results (1) Emodin stimulated luciferase activity controlled by PPRE in dose-dependent manner at concentrations of 0.04 to 180?mol/L in COS-7 cells.The highest value was about 4 folds of control in the cells treated with 90?mol/L emodin (P