This study aims to establish a novel gene-activated matrix that mimics the structure and function of extracellular matrix (ECM-m-GAM). The structure, mechanical property and release profile were also characterized. Firstly, the liposome/DNA lipoplex (LPD) was modified with cell penetrating peptide TAT. The obtained TAT-LPD was then mixed with RGD grafting hyaluronic acid solution. After addition of the matrix metalloproteinase (MMPs) sensitive crosslinker (HS-MMP-SH), hyaluronic acid was crosslinked and TAT-LPD was encapsulated in the subsequently formed hydrogel. As a result, the cell adhesion factor RGD, MMPs sensitive substrate and the efficient gene transfer vector TAT-LPD were all integrated in the hyaluronic acid hydrogel, which was named as ECM-m-GAM. The release profile of DNA from ECM-m-GAM in different release medium was evaluated with PicoGreen kits. The results suggested that the mean diameter of the spherical TAT-LPD was (263.0 ±4.30) nm. TAT-LPD was successfully encapsulated in ECM-m-GAM, which had the typical porous network structure of hydrogels. The mechanical strength of GAM was enhanced with the increasing of hyaluronic acid content. When the content was 4%, the elastic modulus of GAM reached 1 600 Pa. The highly elastic GAM may be suitable for implantation and tissue regeneration. The DNA release showed significant MMPs sensitive property. Especially, the released DNA still existed in form of nanoparticles. Bone marrow mesenchymal stem cells (BMSCs) were successfully transfected with GAM and the green fluorescent protein was expressed. The results have laid a solid foundation for future study of the cell transfection and tissue regeneration.

This study was conducted to investigate the effects of Danlou (correspondence between prescription and syndrome) tablet and Shengmai capsule (non-correspondence between prescription and syndrome) on mini-swine phlegm-stasis syndrome of coronary heart disease (CHD). 24 mini-swines were randomly divided into normal control group, model group, Danlou tablet group (0.24 g·kg^-1) and Shengmai capsule group (0.14 g·kg^-1). Phlegm-stasis syndrome of coronary heart disease was established by high-fat feeding and coronary intervention balloon injury. After 8 weeks of administration, blood lipid levels and blood rheology was detected. Echocardiography was used to examine the changes in heart function, and the extent of infarction was determined by nitro blue tetrazolium (NBT) staining method. The main symptoms, accompanied symptoms, tongue and pulse signs of the coronary heart disease mini-swine with phlegm-stasis syndrome were observed according to the symptom-graded scoring method. The results showed that Danlou tablet decreased serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) (P < 0.05 or P < 0.01), improved the blood rheology (P < 0.01) and cardiac function, increased the left ventricular ejection fraction (EF) and fraction shortening (FS) value (P < 0.05 or P < 0.01), reduced the myocardial infarction area/ventricular area (P < 0.01), significantly lowered the scores of four diagnosis in traditional Chinese medicine (P < 0.05 or P < 0.01). Shengmai capsule improved the hemorheology indices (P < 0.05 or P < 0.01), EF and FS value (P < 0.05), reduced the tongue and pulse signs scores (P < 0.05). However, Shengmai capsule failed to show therapeutic effects on blood lipid metabolism, the myocardial infarction area and primary symptom and syndrome score. The results suggest that as a drug for the treatment of Qi and Yin deficiency syndrome of CHD, Danlou tablet has limited therapeutic effects on phlegm-stasis syndrome of CHD. Only by the prescription correspondence with syndrome, using drug for the treatment of phlegm-stasis syndrome of CHD to treat phlegm-stasis syndrome of CHD, the prescription has a comprehensive therapeutic effect.

This study was designed to investigate the effect of Huangqin Tang (HQT) on gut microbiota of ulcerative colitis (UC) rats, and to explore the relationship between Huangqin Tang and ulcerative colitis and gut microbiota. Fifteen male Wistar rats were randomly divided into control group, trinitrobenzene sulfonic acid (TNBS) group and TNBS + HQT group. The model of UC rats with cell immunoreactivity was made established using the compound method (TNBS and ethanol). After 10 days of administration, 15 fecal samples were collected and total DNA was extracted from the samples to get total DNA. The primers were designed on bacterial 16S rRNA V3-V4 region sequences and Illumina Miseq platform was used for high-throughput sequencing. It was found that the principal component analysis (PCA), the principal co-ordinates analysis (PCoA) and the non-metric multidimensional scale analysis (NMDS) based on the Beta diversity distance showed that there were significant differences in the composition of the gut microbiota among the three groups (P < 0.05). Compared with the control group, the Lactobacillus of the TNBS group was significantly decreased (P < 0.05), and the Lachnospiraceae, Desulfovibrio, Roseburia, Ruminococcaceae were significantly increased (P < 0.05). Compared with the TNBS group, the Lactobacillus in the TNBS + HQT group was significantly increased (P < 0.01), and the Alistipes was significantly decreased (P < 0.05). The study suggests that the Huangqin Tang plays a role in the treatment of ulcerative colitis partially through regulating the structure of the gut microbiota.

Direct-PCR technology was using a 15 minutes heat-lysis step instead of DNA extraction to get DNA templates with small amount of plant materials followed by sensitive PCR process to amplify target genes. In order to facilitate DNA barcoding in medicinal herb identification with Direct-PCR, we collected different tissues from 80 medicinal plants as material to amplify the ITS fragments. Through optimizing the PCR reaction, ITS of 80 plant samples was all successfully amplified. PCR products were sequenced and to do Blast analysis. These results suggest that Direct-PCR would improve the efficiency of DNA barcoding in the application of medicinal herb molecular authentication.

This study was designed to investigate the metabolites of 5F-AMB by human liver microsomes model in vitro. 5F-AMB was added in the reaction mixture to simulate the metabolic process in human hepatocytes in vivo, and then to determine the reaction points and pathways of metabolism by ultra performance liquid chromatography (UPLC) coupled to high resolution mass spectrum (HR-MS). 5F-AMB generated 9 metabolites in total in the human liver microsomes model. Ester hydrolysis, combination of ester hydrolysis and oxidative defluorination, combination of ester hydrolysis and hydroxylation on pentyl chain moiety and combination of ester hydrolysis and hydroxylation on indazole ring moiety reactions were its main metabolic pathways. The method is fast and efficient so that the ester hydrolysis, combination of ester hydrolysis and oxidative defluorination, combination of ester hydrolysis and hydroxylation on pentyl chain moiety and combination of ester hydrolysis and hydroxylation on indazole ring moiety metabolites of 5F-AMB can be used as the suitable and potential biomarkers in the urine samples.

Legumain, a kind of asparaginyl endopeptidase, is overexpressed in highly metastatic and highly aggressive tumor, which can undergo an enzymatic hydrolysis of substrates. We proposed a legumain-responsive functional gold nanoparticle (GNP) drug delivery system (GNPs-A & C), which was consist of Ala-Ala-Asn-Cys-Lys (AK) modified GNPs (GNPs-AK) and 2-cyano-6-aminobenzothiazole (CABT) modified GNPs (GNPs-CABT). In the circulation system, the GNPs-A & C could passively target to the tumor site through the enhanced permeability and retention (EPR) effect. Then the overexpressed legumain specifically cleave the peptide to exposure the 1, 2-thiolamino group, which could take place click reaction with the cyano group of CABT, leading to the aggregation of two GNPs, these aggregates of GNPs with increased size were more likely to retain within tumor site. In vivo fluorescent imaging demonstrated GNPs-A & C could acquire an enhanced accumulation in legumain-overexpressed C6 tumor. Importantly, after tethering DOX, the GNPs-DOX-A & C showed an excellent anti-tumor effect with reduced cardiotoxicity.

To investigated the chemical constituents of the roots of Psidium guajava, we isolated seven compounds by silica gel column chromatography.These include oleanane derivatives, 2 α, 3β, 6β, 23-tetrahydroxylurs-12, 20(30)-dien-28-oic acid β-D-glucopyranoside (1), 2α, 3β, 6β, 23-tetrahydroxylurs-12, 18-dien-28-oic acid β-D-glucopyranoside (2), 2α, 3β, 23-trihydroxylurs-12, 18-dien-28-oic acid β-D-glucopyranoside (3), nigaichigoside F1(4), asiaticoside C (5), 2α, 3β, 6β, 19α, 23-pentahydroxylurs-12, 18-dien-28-oic acid β-Dglucopyranoside (6) and 2α, 3β, 19α, 23-tetrahydroxylurs-12-en-28-oic acid (7).Their structures were elucidated on the basis of spectral analysis with reference to the published data.Compound 1 is brand new, compounds 2-6 were first isolated from this plant.The new compound was evaluated for their cytotoxic activity against human hepatoma Bel 7402 in vitro.The results were expressed as the ratio of inhibiting Bel 7402 cells growth by comparing to untreated cells.The new compound (concentration:25 μmol·L^-1) showed the ratio values of 52.5%.

Due to the characteristics of propofol of high time-varying, and complex compartment model, the traditional method of nonlinear mixed effects modeling (NONMEM) has miscellaneous of variables and plenty of artificial factors in the estimation of propofol. This study was aimed to build a propofol prediction model based on the differential evolution (DE) algorithm and grey model. DE was used to optimize the pa-rameter of multi-variable grey model (MGM) and to build a model of prediction of the plasma concentration of propofol based on the grey model. It was compared with the results of NONMEM algorithm. In conclusion, the median performance error (MDPE) of DE-MGM was -4.6%, while the result of NONMEM is -12.13%. The median absolute performance error (MDAPE) of GA-BP neural network is 13.19%, while that of NONMEM is 23.12%. The experimental results suggest that the new method is suitable to determine the short half-life of anesthesia drug propofol with higher accuracy.

The study is designed to evaluate the protective effect of xanthan gum (XG) injection on cartilage injury in the rabbit osteoarthritis (OA) model induced by anterior cruciate ligament transection (ACLT), and to explore the effect of XG on the expression of caspase-3 and Bax protein in OA cartilage. Sixty male New Zealand white rabbits were randomly divided into 6 groups (n=10) according to random number table method, and one group was selected randomly as the normal control group (control) while the other 5 groups of right knee were used to establish the OA model with ACLT, which were then divided into model group (model), XG-0.6 mg·kg^-1, XG-1.2 mg·kg^-1, XG-2.4 mg·kg^-1 treatment group and sodium hyaluronate (SH-1.2 mg·kg^-1) treatment group according to drug intervention. The knee joint temperature and knee joint width of each group were measured in the course of treatment. After treatment, the macroscopic morphology of rabbit joints in each group was observed. The pathological morphology of articular cartilage of rabbits in each group was observed using HE staining. The expression of Bax and cleaved caspase-3 in the cartilage of rabbits were detected by Western blot. The result shows that XG inhibited the increase in knee joint temperature and knee width caused by OA in a dose-dependent manner. XG improved the morphological abnormalities and tissue injuries of the femoral condyle and tibial plateau caused by OA. Western blot result shows that, compared with the control group, the levels of Bax and cleaved caspase-3 in knee cartilage cells of model group and XG-0.6 mg·kg^-1 group were significantly increased (P < 0.01). However, no significant difference was observed in the levels of Bax and cleaved caspase-3 between the model group and the XG-0.6 mg·kg^-1 group (P>0.05). These two groups are significantly higher than those of XG-1.2 mg·kg^-1 and XG-2.4 mg·kg^-1 (P < 0.01) groups. Meanwhile, no significant difference was observed in the level of cleaved caspase-3 between the knee cartilage in XG-2.4 mg·kg^-1 and XG-1.2 mg·kg^-1 group (P>0.05). The level of Bax in knee cartilage in XG-2.4 mg·kg^-1 group was lower than that of XG-1.2 mg·kg^-1 group (P < 0.05). In conclusion, XG effectively protected cartilage damage in OA, and inhibited the expression of Bax and caspase-3 protein in OA cartilage.

To investigate the effects of metformin on pancreatic β-cell function and its possible mechanism, high fat diet-induced type 2 diabetic C57BL/6J mice were divided into two groups according to fasting blood glucose (FBG), glucose decreasing rate at 40 min of insulin tolerance test, triglycerides (TG), cholesterol (CHO) and body weight (BW). The C57 mice were gavaged with water or metformin for 58 days. β-Cell function was evaluated by oral glucose tolerance test and hyperglycemic clamp. Genes and proteins related to pancreas proliferation, lipid metabolism and endoplasmic reticulum stress were investigated. Compared with the model group, metformin group exhibited a reduction in the body weight (P < 0.01), plasma TG and CHO (P < 0.05), and the area under the curve (AUC) (P < 0.05) of glucose tolerance test. The glucose infusion rate during clamp was improved (P < 0.05) and the fasting insulin level was decreased in the metformin group (P < 0.05). Metformin significantly upregulated the gene expression of pancreatic and duodenal homeobox 1 (Pdx-1, P < 0.01) and liver X receptor β (Lxr-β, P < 0.01). Western blot results showed that, the protein expression of PDX-1 was significantly upregulated (P < 0.01). Endoplasmic reticulum stress related protein of activating transcription factor 4 (ATF4, P < 0.001) and C/EBP homologous protein (CHOP, P < 0.05) were also down-regulated. These results suggest that metformin could improve the insulin secretion function of type 2 diabetic C57BL/6J mice. The mechanism of the action may rely on its improvement of pancreas cell proliferation, lipid metabolism and amelioration of endoplasmic reticulum stress.