OBJECTIVETo observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers, such as alkaline phosphatase, Cbf(alpha)l and type I collagen, and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head.

METHODSFifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs, then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood, cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group, model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro, and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week,the expressions of alkaline phosphatase, Cbf(alpha)l and type I collagen mRNA were detected.

RESULTSThe alkaline phosphatase activity was significantly decreased in BMSCs of the model group as compared with the blank group (P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly increased in BMSCs of each HSYA group as compared with the model group (P < 0.05 or P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also increased significantly (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chalcone ; analogs & derivatives ; chemistry ; pharmacology ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor alpha Subunits ; genetics ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rabbits

Abscess ; Cysts ; Female ; Humans ; Maxillary Sinus ; Tooth Diseases ; Young Adult

Aim To evaluate the effects of aspirin on the expression of inflammatory proteins induced by oxidized low-density lipoprotein(ox-LDL) in human vascular endothelial cells (HUVECs). Methods HUVECs were stimulated with different concentrations of ox-LDL. The expression of inflammatory proteins was detected by Western blot.Intracellular ROS generation was measured by flow cytometry using perexide-sensitive flurscent probe 2′, 7′-dichrofluorescein diacetate(DCFH-DA).Results ① Aspirin inhibited COX-2 expression induced by ox-LDL. Cells were preincubated with 2.5 mmol?L-1, 5 mmol?L-1 of aspirin or without any treatment for 30 min and then stimulated by 0.3 g?L-1 ox-LDL for 16 h, COX-2 expression was reduced by treating of aspirin.COX-2 expression was enhanced after the stimulation with ox-LDL, and aspirin inhibited the increasing.② Aspirin suppressed ICAM-1 expression induced by ox-LDL in HUVECs. ICAM-1 expression was increased by ox-LDL stimulation for 16 h, and aspirin significantly down-regulated the expression. Similar results were obtained by immunofluorescence.③ Aspirin partially reduced ROS production induced by ox-LDL in HUVECs. After stimulation with 0.3 g?L-1 ox-LDL for 16 h, the intracellular level of ROS was increased, however, aspirin failed to fully inhibit the phenomenon.Conclusion Nonsteroidal anti-inflammatory drugs (NSAID) aspirin significantly down-regulated the expression of COX-2 and ICAM-1 induced by ox-LDL.The results suggested that aspirin could reduce the inflammation responses mediated by ox-LDL on HUVECs in atherosclerosis.

Objective To optimize the therapeutic dose and time window of picroside Ⅱ in treating cerebral ischemic injury in rats by orthogonal test. Methods The forebrain ischemia models were established by bilateral common carotid artery occlusion (BCCAO) method. The successful models were randomly grouped according to orthogonal experimental design and treated by injecting picroside Ⅱintraperitoneally at different ischemic time with different doses. The concentrations of MDA, NO and H2O2 in serum and brain tissue were respectively determined by thiobarbituric acid assay, nitratase reductase assay and chemiluminescence immunoassay. Results The optimized composition of the therapeutic dose and time window of picroside Ⅱ in cerebral ischemic injury were ischemia 1.5 h with 10 mg/kg, 1.5 h with 20 mg/kg and 1.5 h with 10 mg/kg body weight according to the expressions of MDA, NO and H2O2 in serum, and ischemia 1.5 h with 10 mg/kg, 1.5 h with 20 mg/kg and 1.5 h with 20 mg/kg body weight according to the expressions of MDA, NO and H2O2 in brain tissue. Conclusion On the basis of the principle of lowest therapeutic dose with longest time window, the optimized composition of the therapeutic dose and time window in cerebral ischemic injury is injecting picroside Ⅱ intraperitoneally with 10-20 mg/kg body weight at ischemia 1.5 h.

ObjectiveTo investigate the inhibitory effect of 1-methyl-tryptophan (1-MT) on transplanted hepatocellular carcinoma in mice.MethodsHuman hepatocellular carcinoma subcutaneous tumor models were established in mice,and the mice were divided into hepG2 group,empty plasmid group,indoleamine 2,3-dioxygenase (IDO) saline group,IDO) 5-fluoropyrimidine (5-FU) group,IDO 1-MT group,and the group combining IDO 1-MT with 5-FU treatment (n=8 in each group).The tumor growth,tumor volume and pathological examination were observed and the expression of IDO in tumor tissues was determined by immunohistochemistry.ResultsCompared with hepG2 and the empty vector saline groups,IDO saline group had bigger tumor,faster growth,and the differences were statistically significant (P＜0.05).Compared with IDO saline group,5-FU group,1-MT group and combination treatment groups showed smaller tumor volume and weight,and the tumor inhibitory rates were 86.54％,79.95％,94.46％,respectively.There were significant differences between these groups (P＜0.05).However,there were no significant differences in tumor volumes between 5-FU group and 1-MT group (P＞0.05).HE pathological observation of ceils in each treatment group showed reduced density,increased necrotic area and significant decrease in peripheral blood alpha-fetoprotein (P＜0.05).ConclusionsIDO can promote the growth of liver cancer cells involved in immune escape.1-MT can inhibit the transplanted tumor growth in mice,and therefore may enhance the chemotherapeutic efficacy.

Objective:To explore the methods and ideas for developing pharmaceutical care in clinical practice. Methods: The pharmaceutical care performed by clinical pharmacists and the therapeutic scheme assisted by clinical pharmacists for one patient with adult purulent meningitis were analyzed retrospectively. Results and Conclusion:Through selection of anti-infective agents, treatment of adverse drug reactions and assessment of patients’ economic capacity,clinical pharmacists help provide reasonable medication to im-prove therapeutic efficacy, safety and economy.

Objective:To investigate the clinicopathological features of gastric adenocarcinoma of fundic gland type (GA-FG).Methods:A total of 12 patients, including 7 cases treated with endoscopic submucosal dissection (ESD), were diagnosed as having GA-FG in Nanjing Drum Tower Hospital from January 2018 to August 2019. Morphological changes were analyzed by reviewing endoscopic and pathological results. Patients were followed up after definitive diagnosis.Results:The clinical symptoms of patients with GA-FG were nonspecific. No Helicobacter pylori infection was identified. The lesions were found in the non-atrophic gastric mucosa of the upper 1/3 portion in 10 cases and middle 1/3 portion in 2 cases. Endoscopically, the most common features were whitish color (9 cases), and all lesions diameter≤1 cm. Their macroscopic types were classified as 0-Ⅰ (2 cases), 0-Ⅱa (9 cases) and 0-Ⅱc (1 case) respectively. All lesions had sharp boundary, with branching dilated blood vessels on the surface. Five in 7 cases who were treated with ESD showed submucosal invasion. Immunohistochemically, 9 cases were classified as the chief cell type , 3 as the mixed type, 11 MUC6 positive, 4 MUC5AC positive, 2 MUC2 positive, and 3 CD10 positive. P53 was detected in all 12 cases, and 9 cases had low Ki-67 staining index (<10%). The mean time of follow-up was 11 months, and 11 patients survived. Conclusion:GA-FG should be taken into consideration when the polyps are found in the upper part of the stomach, with whitish color, and branch dilated blood vessels on the surface. Excellent clinical outcomes can be achieved for GA-FG patients with ESD.

Objective To evaluate two methods of selecting region of interest (ROI)to measure the apparent diffusion coefficient (ADC)of breast lesions on diffusion-weighted imaging (DWI).Methods MRI findings (including plain MRI,dynamic contrast-en-hanced MRI and DWI)of 5 3 cases with breast mass confirmed by pathology were analyzed retrospectively.Two observers independ-ently measured the ADC of breast lesions by the method of fixed ROI with the size of 10 mm2 ±2 mm2 and the routine method.The mean ADCs of benign and malignant breast lesions were compared between two methods and two observers by using paired samples T-test.The values of ADC diagnosing breast cancer were compared between the two methods by using the receiver operating charac-teristic (ROC)curve.Results Among 62 lesions of 53 cases,33 lesions were malignant,and 29 lesions were benign.For the first observer,the mean ADCs of benign and malignant lesions were (1.647±0.359)×10-3 mm2/s and (1.136±0.227)×10-3 mm2/s measured by the fixed ROI method,respectively.The area under the ROC was 0.897.The mean ADCs of benign and malignant le-sions were (1.603±0.415)×10-3mm2/s and (1.251±0.237)×10-3 mm2/s measured by the routine method,respectively.The area under the ROC was 0.768.For another observer,the mean ADCs of benign and malignant lesions were (1.647±0.357)× 10-3 mm2/s and (1.130±0.250)×10-3 mm2/s measured by the fixed ROI method,respectively.The mean ADCs of benign and malignant lesions were (1.590± 0.390)×10-3 mm2/s and (1.250±0.237)×10-3 mm2/s measured by the routine method,respec-tively.There was no significant difference for the mean ADC of benign lesions between the two methods.However,there was sig-nificant difference for the mean ADC of malignant lesions between the two methods.The diagnostic value of ADC measured by the fixing method was better than that of the routine method (P=0.008).There were no significant differences for ADCs of benign and malignant lesions between the two observers.Conclusion DWI has an important value for differential diagnosis of benign and malignant breast lesions.The measurement of ADC by the fixed and smaller ROI is better than that by the routine ROI for dis-tinguishing malignant from benign breast lesions.

WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western ; Cloning, Molecular ; DNA-Binding Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; chemistry ; genetics ; metabolism ; Molecular Weight ; Plant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Salvia miltiorrhiza ; genetics

This study is to investigate the protective effect of longistyline A against corticosterone-induced neurotoxicity in PC12 cells. While PC12 cells were exposed to 100 micromol x L(-1) corticosterone for 48 h, cell survival rate was reduced and lactate dehydrogenase (LDH) release increased. In parallel, corticosterone caused significant elevations of DNA fragmentation, [Ca2+]i and caspase-3 activity. However, when the PC12 cells were incubated with longistyline A (4.0, 8.0 and 16.0 micromol x L(-1)) in the presence of 100 micromol x L(-1) corticosterone for 48 h, the effects were evidently alleviated, but dose-dependent manner was not obvious. In summary, longistyline A could generate a neuroprotective effect against corticosterone-induced neurotoxicity in PC12 cells possibly by decreasing [Ca2+]i and caspase-3 activity.
Animals ; Cajanus ; chemistry ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Corticosterone ; toxicity ; DNA Fragmentation ; drug effects ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Neuroprotective Agents ; isolation & purification ; pharmacology ; PC12 Cells ; Phenols ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats