To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.
Cellulase ; metabolism ; Drug Compounding ; methods ; Flavonoids ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Temperature ; Time Factors

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Morphogenetic Protein 4 ; metabolism ; Carrier Proteins ; pharmacology ; Liver ; metabolism ; Liver Regeneration ; Male ; Rats ; Rats, Wistar

AIM:To investigate the effect of phosphatidylinostiol 3-kinase ( PI3K) inhibitor GDC-0032 on cell viability, cell cycle and DNA damage in human glioma U 251 cells.METHODS:The cell viability was analyzed by MTT assay.The cell cycle distribution of U251 cells was examined by flow cytometry .The protein expression was examined by Western blot.The expression and localization of γ-H2AX were determined by laser-scanning confocal microscopy .RE-SULTS:GDC-0032 inhibited the cell viability in a dose-dependent manner .U251 cells showed G 1 phase arrest accompa-nied with upregulation of p 27 protein after exposure to GDC-0032, while the expression of cell division cycle protein 2 (Cdc2) was inhibited.GDC-0032 treatment increased the formation of γ-H2AX foci and histone H2AX phosphorylation in the U251 cells.In addition, GDC-0032 upregulated the phosphorylation levels of mitogen-activated protein kinases , inclu-ding extracellular signal-regulated kinase ( ERK) and c-Jun N-terminal kinase ( JNK) , in the glioma cells , while the ex-pression of survivin was inhibited .CONCLUSION:GDC-0032 inhibits U251 cell growth by inhibition of cell viability and cell cycle progression.GDC-0032 also induces DNA damage of U251 cells.The anticancer activity of GDC-0032 is associ-ated with increasing the activity of ERK and JNK and downregulating the expression of survivin .

OBJECTIVETo study the curative effect of Zhiyang Pingfu Liquid (ZPL) in treating epidermal growth factor receptor inhibitors (EGFRIs) associated adverse reactions of the skin.

METHODSAll 54 patients with pathologically confirmed malignant tumor had EGFRIs induced adverse reactions of the skin to various degrees. ZPL was externally applied for them all, once or twice per day, 14 days consisting of one therapeutic course. Changes of adverse skin reactions, time for symptoms relief, adverse skin reaction types suitable for ZPL were observed before and after treatment.

RESULTSEGFRIs associated skin adverse reactions were improved to various degrees after they used ZPL. The shortest symptoms relief time was 1 day while the longest was 12 days, with an average of 6.93 days and the median time 7 days. Compared with before treatment, itching, rash/scaling, acne/acneform eruptions were obviously improved (P < 0.05).

CONCLUSIONZPL could alleviate EGFRls associated adverse skin reactions, especially showed better effect on itching, rash/scaling, acne/acneform eruptions.

Antineoplastic Agents ; adverse effects ; Biomedical Research ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Exanthema ; chemically induced ; Humans ; Neoplasms ; drug therapy ; Pruritus ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Skin ; drug effects ; Skin Diseases ; drug therapy

Protein disulfide isomerase-related protein A (PRPA) was highly expressed (about 34%) in Escherichia coli by inserting the whole PRPA cDNA into the vector pET23b. After expression, the purified protein was acquired through ammonium fractional precipitation and Bio-Rex 70 chromatography. PRPA shows low disulfide isomerase activity (only about 1/250 of that of hPDI), decreases the reactivation yield of denatured and reduced lysozyme either in redox and non-redox Hepes buffer or redox PBS buffer and facilitates the aggregation of denatured and reduced lysozyme. Fluorescence spectra of PRPA indicate that PRPA has more hydrophobic groups at surface than that of hPDI, and which can be used to explain why PRPA has anti-chaperone activity during the refolding of denatured and reduced lysozyme.
Cloning, Molecular ; Fungal Proteins ; chemistry ; genetics ; isolation & purification ; Muramidase ; chemistry ; Plasmids ; Protein Folding ; Recombinant Proteins ; biosynthesis ; isolation & purification ; Spectrometry, Fluorescence

OBJECTIVETo investigate the effects of rabbit limbs ischemia/reperfusion on myocardial necrosis and apoptosis in vivo.

METHODSThirty-six healthy new zealand rabbits were randomly divided into 3 groups: (1) Sham group; (2) I/R(Ischemia/reperfusion) group; (3) RPostC (remote postconditioning) group. The activity of blood serum creatine kinase (CK) and lactate dehydrogenase (LDH) were measured at baseline, the end of ischemia after 60 min and 120 min of reperfusion respectively. The extent of myocardial ischemia and the scope of myocardial infarction were assessed by evans blue and Triphenyl tetrazolium chloride (TTC). The myocardial cell's apoptosis at the area of myocardial ischemia was estimated by Tunel. Protein expression of caspase-3, Bcl-2 and Bax in myocardial ischemic area were analyzed with the method of immunohistochemistry.

RESULTSCompared with I/R group, the myocardial infarct size and the CK activity were significantly reduced in RPostC group. The Tunel positive index of RPostC group in ischemic myocardium was significantly lower than that in I/R group (21.79% +/- 1.07% vs 35.81% +/- 1.10%, P < 0.05). Caspase-3 positive cells index was calculated with randomly selected five regions in each slide and then the positive cells in per hundred cells were calculated. The RPostC group of caspase-3 positive cells was significantly lower than that in I/ R group(25.03% +/- 1.16% as 39% +/- 2.43%, P < 0.05). Compared with the sham group, the Bax protein expression index and the Bcl-2 protein expression index of I/R group and RPostC group were increased. The Bax/Bcl-2 ratio of RPostC group decreased, while it was increased in I/R. Compared with the I/R group, the Bax protein expression and Bax/Bcl-2 ratio of RPostC group significantly reduced, but the expression index of Bcl-2 ratio was significantly increased, the differences were statistically significant.

CONCLUSIONLimbs ischemia/postconditioning could significantly reduce necrosis and apoptosis of ischemia/reperfusion myocardium. The mechanism of reducing the myocardial cell apoptosis may have relation to inhibiting the activation of pro-apoptotic gene caspase-3 and increased expression of Bcl-2.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Creatine Kinase ; blood ; Ischemic Postconditioning ; L-Lactate Dehydrogenase ; blood ; Lower Extremity ; Male ; Muscle, Skeletal ; blood supply ; Myocardial Reperfusion Injury ; pathology ; Necrosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the effect of transcatheter arterial chemoembolization(TACE) using As_2O_3 and Lipiodol on the growth and metastasis of the implanted hepatic tumor in rabbits and the correlation of metastasis with angiogenesis of the residual tumor.Methods Forty-eight New Zealand white rabbits were randomly divided into four groups and VX_2 carcinoma was implanted in the left lobes of the livers.Two weeks later,a catheter was inserted into the hepatic artery and infusion was performed via the hepatic artery using physiological saline(group A),Lipiodol(group B),ADM-Lipiodol(group C),and As_2O_3-Lipiodol(group D),respectively.One week after the treatment,the value of microvessel density (MVD)of tumors(samples got by biopsy)was examined by immunohistochemistry.Three weeks after the treatment,the volume and necrotic area of the implanted tumor were measured.The metastases in the liver, lungs and other organs were recorded.Results One week after the treatment,the value of MVD of the tumorswas(21.8?5.3),(23.4?3.9),(22.4?4.5),and(14.3?3.4)/400 power LM(F= 11.246,P=0.000).Three weeks after the treatment,the mean volume of the implanted tumor was (35.5?7.1),(21.2?8.3),(20.7?9.1),and(11.8?3.7)cm~3(F=21.203,P=0.0000)in groups A,B,C and D,respectively.There was significant difference between group D and group B(q= 4.398,P

Objective To analyze the amount and tendency of methyl methacrylate(MMA) released from autopolymerized denture base polymer (self-curing resin) during processing using time-of-flight mass spectrometer(TOF-MS). Methods Self-curing resin was mixed in the container using a ratio of 2 g of powder to 1 g of liquid in accordance with the manufacturer's instructions for 40 s as a specimen. The amount of MMA released from the specimen was continuously monitored and simultaneously recorded every minute by TOF-MS since immediately after mixing. A total of five specimens were monitored. Results The amount of MMA increased dramatically at 11 min[(45.2 ±3.5) mg/L] after mixing, and reached the highest level at 13 min[(228.9 ±22.6) mg/L], then become stable at 23 min[(8.8 ±2. 3) mg/L] after mixing. Conclusions The releasing tendency of MMA could be analyzed accurately with continuously monitoring during processing. The amount of MMA released from self-curing resin changed rapidly and the processing was complicated and changeful.

Objective To develop the finite element model of six-year-old child occupant lower extremity with higher biofidelity and validate the model of knee joints,as well as analyze the biomechanical responses of growth plate under frontal impact load and injury mechanisms of the knee joint by using this model.Methods The sixyear-old child occupant lower extremity with growth plate was modeled based on children's anatomy and CT images,and corresponding material properties of the lower extremity model were assigned.The model was validated according to biomechanical experiments by Kerrigan et aL and Haut et aL and then was used to analyze the injury results of growth plate with different material properties.Results The model validation was qualified by comparing the curves from the experimental and simulation results.The growth plates at knee regions could change injury patterns of the child occupant lower extremity fracture.The material properties of growth plate could affect threshold of axial damage of the femur as well as relative position of the fracture.Conclusions The validated model can be used for related study and application on biomechanical responses and injury mechanisms of sixyear-old child occupant lower extremities.

Objective: To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro. Methods: Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion. Results: ①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients' T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05). Conclusion: Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.
Antigens, CD19 ; Cell Proliferation ; Humans ; Leukocytes, Mononuclear ; Nivolumab/pharmacology* ; Programmed Cell Death 1 Receptor ; Receptors, Antigen, T-Cell ; Receptors, Chimeric Antigen ; T-Lymphocytes