New observations on the effect of camellia oil on fatty liver disease in rats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Camellia oil has become an important plant oil in China in recent years, but its effects on non-alcoholic fatty liver disease (NAFLD) have not been documented. In this study, the effects of camellia oil, soybean oil, and olive oil on NAFLD were evaluated by analyzing the fatty acid profiles of the plant oils, the serum lipids and lipoproteins of rats fed different oils, and by cytological and ultrastructural observation of the rats' hepatocytes. Analysis of fatty acid profiles showed that the polyunsaturated fatty acid (PUFA) n-6/n-3 ratio was 33.33 in camellia oil, 12.50 in olive oil, and 7.69 in soybean oil. Analyses of serum lipids and lipoproteins of rats showed that the levels of total cholesterol and low-density lipoprotein cholesterol in a camellia oil-fed group (COFG) were lower than those in an olive oil-fed group (OOFG) and higher than those in a soybean oil-fed group (SOFG). However, only the difference in total cholesterol between the COFG and SOFG was statistically significant. Cytological observation showed that the degree of lipid droplet (LD) accumulation in the hepatocytes in the COFG was lower than that in the OOFG, but higher than that in the SOFG. Ultrastructural analysis revealed that the size and number of the LDs in the hepatocytes of rats fed each of the three types of oil were related to the degree of damage to organelles, including the positions of nuclei and the integrity of mitochondria and endoplasmic reticulum. The results revealed that the effect of camellia oil on NAFLD in rats was greater than that of soybean oil, but less than that of olive oil. Although the overall trend was that among the three oil diets, those with a lower n-6/n-3 ratio were associated with a lower risk of NAFLD, and the effect of camellia oil on NAFLD was not entirely related to the n-6/n-3 ratio and may have involved other factors. This provides new insights into the effect of oil diets on NAFLD.

New observations on the effect of camellia oil on fatty liver disease in rats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Camellia oil has become an important plant oil in China in recent years, but its effects on non-alcoholic fatty liver disease (NAFLD) have not been documented. In this study, the effects of camellia oil, soybean oil, and olive oil on NAFLD were evaluated by analyzing the fatty acid profiles of the plant oils, the serum lipids and lipoproteins of rats fed different oils, and by cytological and ultrastructural observation of the rats’ hepatocytes. Analysis of fatty acid profiles showed that the polyunsaturated fatty acid (PUFA) n-6/n-3 ratio was 33.33 in camellia oil, 12.50 in olive oil, and 7.69 in soybean oil. Analyses of serum lipids and lipoproteins of rats showed that the levels of total cholesterol and low-density lipoprotein cholesterol in a camellia oil-fed group (COFG) were lower than those in an olive oil-fed group (OOFG) and higher than those in a soybean oil-fed group (SOFG). However, only the difference in total cholesterol between the COFG and SOFG was statistically significant. Cytological observation showed that the degree of lipid droplet (LD) accumulation in the hepatocytes in the COFG was lower than that in the OOFG, but higher than that in the SOFG. Ultrastructural analysis revealed that the size and number of the LDs in the hepatocytes of rats fed each of the three types of oil were related to the degree of damage to organelles, including the positions of nuclei and the integrity of mitochondria and endoplasmic reticulum. The results revealed that the effect of camellia oil on NAFLD in rats was greater than that of soybean oil, but less than that of olive oil. Although the overall trend was that among the three oil diets, those with a lower n-6/n-3 ratio were associated with a lower risk of NAFLD, and the effect of camellia oil on NAFLD was not entirely related to the n-6/n-3 ratio and may have involved other factors. This provides new insights into the effect of oil diets on NAFLD.

Study on transdermal permeation of ethanol extracts from roots of Gentiana crassicaulis in vitro and in vivo / 中草药

Objective: To establish a determination method for kinds of chemical components of iridoids in the roots of Gentiana crassicaulis in transdermal absorption liquid, and research its transdermal permeability, so as to provide scientific basis for transdermal delivery system, clinical medication and reform of the traditional forms of G. crassicaulis. Methods: Based on the results of the previous investigation, in this paper, using the roots of G. crassicaulis as the research subject, a certain concentration solution of G. crassicaulis extract was prepared by the alcohol extraction method. Three kinds of common penetration enhancers, azone, borneol and propylene glycol were used. The effects of single penetration enhancer and dual compound penetration enhancers on the transdermal penetration of five kinds of chemical components of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside in vitro and the three kinds of chemical components of gentiopicroside, loganic acid and swertimarin in vivo were quantitatively studied by the method of HPLC to investigate the transdermal permeability of G. crassicaulis extract in mice skin model. Results: According to the experimental results, compared to the control group, penetration enhancers significantly increased the absorption of five chemical components of G. crassicaulis in vitro. Transdermal absorption rates (J) of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside were 12.306 0, 1.248 8, 4.187 5, 153.030 0 and 5.012 6 μg/(cm2∙h), respectively. The transdermal enhancer effects of A (5% azone), B (5% borneol), C (5% propylene glycol), A + B (2.5% azone and 2.5% borneol), A + C (2.5% azone and 2.5% propylene glycol), A + C (2.5% borneol and 2.5% propylene glycol) were 9.73, 2.57, 13.94, 15.92 and 8.08 times faster than the control group, respectively. Among these, the group of A + C had a marked osmotic enhancer effect in vitro. In comparison with the control group, the in vivo percutaneous penetration test indicated that the dual compound penetration enhancers of 2.5% azone and 2.5% propylene glycol had a marked effect for the permeability enhancement. Conclusion: This study showed azone and propylene glycol significantly promoted the percutaneous penetration effect of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside of G. crassicaulis, and this study laid a foundation for the quality control of percutaneous drug delivery preparation of G. crassicaulis.

Anti-senescence effect and molecular mechanism of the major royal jelly proteins on human embryonic lung fibroblast (HFL-I) cell line / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Royal jelly (RJ) from honeybee has been widely used as a health promotion supplement. The major royal jelly proteins (MRJPs) have been identified as the functional component of RJ. However, the question of whether MRJPs have anti-senescence activity for human cells remains. Human embryonic lung fibroblast (HFL-I) cells were cultured in media containing no MRJPs (A), MRJPs at 0.1 mg/ml (B), 0.2 mg/ml (C), or 0.3 mg/ml (D), or bovine serum albumin (BSA) at 0.2 mg/ml (E). The mean population doubling levels of cells in media B, C, D, and E were increased by 12.4%, 31.2%, 24.0%, and 10.4%, respectively, compared with that in medium A. The cells in medium C also exhibited the highest relative proliferation activity, the lowest senescence, and the longest telomeres. Moreover, MRJPs up-regulated the expression of superoxide dismutase-1 (SOD1) and down-regulated the expression of mammalian target of rapamycin (MTOR), catenin beta like-1 (CTNNB1), and tumor protein p53 (TP53). Raman spectra analysis showed that there were two unique bands related to DNA synthesis materials, amide carbonyl group vibrations and aromatic hydrogens. These results suggest that MRJPs possess anti-senescence activity for the HFL-I cell line, and provide new knowledge illustrating the molecular mechanism of MRJPs as anti-senescence factors.

Initial study on preparation and properties of porous Bletilla striata polysaccharides gum / 中草药

Obiective Guided by the thought of unification of drug and adjuvant, Bletilla striata polysaccharides gum (BSPG) was got from traditional Chinese medicine of Bletilla striata. Methods In this research, the porous BSPG was prepared and its functional properties in solid and gel states were characterized by using infrared-assisted extraction, ethanol fractionation precipitation, freeze drying, and texture analyzer. BSPG40, BSPG60, and BSPG80 were fractionated by repeated precipitation method, using 40%, 60%,and 80% ethyl alcohol as precipitant and these polysaccharides content were determined with Phenylhydrate-Sulfuric acid. Octadecanol as the adjunct and the porous BSPG as the material, porous and non-porous tablets were prepared by direct compression method. The capabilities of floating, water absorption, expansibility, and the degradation performance were measured in artificial gastric juice to give an initial research on the feasibility that the porous BSPG can be used as adjuvant. Results It showed that the total polysaccharide content of BSPG, BSPG40, BSPG60, and BSPG80 was 64.15%, 69.33%, 57.64%, and 42.83%, respectively and the yield of BSPG40, BSPG60 and BSPG80 account for 83.25%, 12.16%, and 4.49%. BSPG80 is too little to get and exhibits typical “weak gel” properties, so properties of BSPG, BSPG40, BSPG60 were tested. By freeze drying, all samples were loose and porous. Hardness increased as the concentrations increased, and at equal concentrations, the hardness of BSPG, BSPG40, and BSPG60 gradually increased but filling power gradually decreased as the hardness increased. Elasticity of BSPG, BSPG40, and BSPG60 is better at the concentration of 15, 20, and 20 mg/mL respectively. While gel strength and adhesion of BSPG, BSPG40, and BSPG60 was gradually weaken. This study’s preliminary experiments indicated that tablets of porous BSPG had good ability of floating, water absorption, expansibility and the degradation performance. Conclusion The experiment results showed that different concentration of ethanol fractionation precipitation exercises a great influence on the texture properties and at the same time, different concentration of BSPG which obtained by the same concentration of ethanol fractionation precipitated has different texture properties and low concentrations of ethanol fractionation precipitated had better performance of hardness, filling power, gel strength, adhesion and played a better role in pharmacy. Besides, porous BSPG is a potential excipient of floating to prolong the gastric retention time. Therefore, this study provided important theoretical evidence and made great significance to future exploration and development of the new excipient of porous BSPG.

Research on quality control method for Juhuang Oral Liquid / 中草药

Objective: To establish the quality control method for Juhuang Oral Liquid (JOL) based on the qualitative and quantitative analysis methods. Methods: TLC was used to identify Lycii Fructus and Polygonati Rhizoma in JOL. Phenol-sulfuric acid method was used for the detemination of total polysaccharide, HPLC was used to simultaneously determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, coffeic acid, galuteolin, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid. The Odyssil C18 (250 mm × 4.6 mm, 5 μm) column was used. The mobile phase was acetonitrile-0.05% phosphoric acid solution in gradient elution, at the flow rate of 1.0 mL/min, the detection wavelength was set at 325 nm. Results: The identified characteristics of Lycii Fructus and Polygonati Rhizoma by TLC were distinct and the spots were clear, without interference and easy to recognize. When phenol-sulfuric acid method was used to determinate the total polysaccharide content, the regression equation was A=0.057 8 C + 0.032 5 (r=0.999 1, n=8). The linear with its absorbance in the range of 26.15-196.13 mg/mL, the average recoveries was 99.82%. The average content of the total polysaccharide in JOL from six batches is 157.74-166.49 mg/mL. When RP-HPLC was used to determine the eight chemical components, the compounds showed a good linearity(r ≥ 0.999 8) in the determination range. The average recoveries were between 99.18%-101.06% with RSDs between 0.79%-1.91%. The average content from the six batches showed neochlorogenic acid 102.9-142.6 μg/mL, chlorogenic acid 488.6-563.8 μg/mL, cryptochlorogenic acid 58.9-71.6 μg/mL, coffeic acid 240.7-326.1 μg/mL, galuteolin 228.6-302.7 μg/mL, 3,4-dicaffeoylquinic acid 398.0-485.4 μg/mL, 3,5-dicaffeoylquinic acid 184.1-203.1 μg/mL, and 4,5-dicaffeoylquinic acid 476.2-561.8 μg/mL. Conclusion: TLC identification method is simple. The determination method of total polysaccharide and chemical components is simple, accurate, and with a good repeatability. It can be used for the quality control of JOL.

Content determination of twelve major components in Tibetan medicine Zuozhu Daxi by UPLC / 中国中药杂志

A quantitative analytical method of ultra-high performance liquid chromatography (UPLC) was developed for simultaneously determining twelve components in Tibetan medicine Zuozhu Daxi. SIMPCA 12.0 software was used a principal component analysis PCA) and partial small squares analysis (PLSD-DA) on the twelve components in 10 batches from four pharmaceutical factories. Acquity UPLC BEH C15 column (2.1 mm x 100 mm, 1.7 µm) was adopted at the column temperature of 35 °C and eluted with acetonitrile (A) -0.05% phosphate acid solution (B) as the mobile phase with a flow rate of 0. 3 mL · min(-1). The injection volume was 1 µL. The detection wavelengths were set at 210 nm for alantolactone, isoalantolactone and oleanolic; 260 nm for trychnine and brucine; 288 nm for protopine; 306 nm for protopine, resveratrol and piperine; 370 nm for quercetin and isorhamnetin. The results showed a good separation among index components, with a good linearity relationship (R2 = 0.999 6) within the selected concentration range. The average sample recovery rates ranged between 99.44%-101.8%, with RSD between 0.37%-1.7%, indicating the method is rapid and accurate with a good repeatability and stability. The PCA and PLSD-DA analysis on the sample determination results revealed a great difference among samples from different pharmaceutical factories. The twelve components included in this study contributed significantly to the quantitative determination of intrinsic quality of Zuozhu Daxi. The UPLC established for to the quantitative determination of the twelve components can provide scientific basis for the comprehensive quality evaluation of Zuozhu Daxi.

Distribution and efficiency of recombinant adenovirus mediated human stem cell leukemia gene transfer in mice with interstitial cells of Cajal loss / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the construction of the green fluorescent protein (GFP) labeled recombinant adenovirus containing human stem cell leukemia (hSCL) and its distribution and efficiency in mice with interstitial cells of Cajal (ICC) loss.</p><p><b>METHODS</b>The recombinant adenovirus Ad-GFP/SCL was constructed by Ad-Easy system based on the homologous recombination in bacteria, then 1.6 x 10(9) PFU of recombinant adenoviruses were injected into Balb/c mice with ICC loss via the tail vein. In vivo distribution and efficiency of recombinant adenoviruses mediated hSCL were observed by GFP under the fluorescent microscope at different phases. The expression of SCL gene was measured by RT-PCR method. The damages were observed in different organs by HE staining.</p><p><b>RESULTS</b>The recombinant adenovirus containing hSCL was quickly constructed by homologous recombination in bacteria using Ad-Easy system. Under the fluorescent microscope, GFP was revealed in heart, lung, liver, spleen, kidney, small intestine and large intestine of mice with ICC loss at different phases. No obvious damages were observed in various visceral organs by HE staining. RT-PCR showed SCL mRNA expression in various visceral organs at different levels.</p><p><b>CONCLUSIONS</b>Construction of adenovirus vector by the homologous recombination in bacteria is an efficient and time saving method, and a high titer of adenovirus is able to mediate the safe and stable expression of SCL gene in mice with ICC loss. This finding will make further gene therapy in mice with STC possible.</p>