Objective:To'synthesize BODIPY-FL-labeled phenylephrine(BODIPY-FL-PE)and deter-mine its biological activity.Methods:Condensation of BODIPY-FL(green fluorescence dye)and phe-nylephrine(?_1-adrenoceptor agonist)was performed by adding dicyclohexylcarbodiimide(DCC)in thepresence of absolute tetrahydrofuran(THF).The reaction occurred in absolutely oxygen and water condi-tion at room temperature.The crude product was separated and purified by thin-layer chromatography(TLC).The structure of BODIPY-FL-PE was characterized by TLC and mass spectrometry(MS).Itspharmabiological activity was determined by Western blot.Results:BODIPY-FL-PE,the target mole-cule,was synthesized and its structure was identified by using ultra-violet spectrometry(UV)and MS.The result of Western blot indicated that ?_1-adrenoceptor(?_1-AR)induced ERK phosphorylation wasconfirmed in both BODIPY-FL-PE and PE treated groups.Conclusion:The synthesized BODIPY-FL-PEhas pharmacological activity that could activate ?_1-AR.Visualization of AR behaviors could be achievedby tracing the trajectories of BODIPY-FL-PE labeled AR.It might be a promising tool for investigatingdynamic behaviors of AR in living cells.

·AIM: To investigate whether Erigeron Breviscapus (vant) Hand-Mazz (EBHM) EBHM has neuroprotective effect against N-methyl-D-aspartate (NMDA)-induced neuron death in retinal ganglion cell layer (RGCL).· METHODS: 60 healthy SD rats were randomly divided into four groups. 6 animals were normal control group (group A). The others were divided as group B (EBHM group), group C (normal saline+NMDA group) and group D (EBHM + NMDA group). Each group had 18 rats.10nmol NMDA was intravitreally injected to induce partial damage of the neurons in RGCL in the right eyes of Groups C and D. Same volume PBS was intravitreally injected into the left eyes as self-control. Groups B and D were pre-treated intraperitoneally with 6g/L EBHM solution at a dose of 150mg/kg body weight/day seven days before and after NMDA treatment. Group C were administrated intraperitoneally with 9g/L normal saline at the same time of EBHM injection. Rats were sacrificed at 4,7,14d after NMDA treatment. Flat whole retinas were stained with 5g/L cresyl violet and neuron counting in RGCL from both eyes were observed. Each subgroup had 6 rats.· RESULTS: There was no significant difference of neuron counting in RGCL between the right eye and the left eye in group A (P=0.200). There was no significant difference between normal control group and EBHM group either in the right eyes or in the left eyes at 4, 7 and 14 d respectively after intravitreal injection of 10nmol NMDA in group C and group D. (P=0.636, P=0.193). Neuron counting of RGCL in group C and D was significantly decreased in the NMDA-treated eyes at 4, 7 and 14d after intravitreal injection (P＜0.001). There was no significant difference between self-control eyes group and normai control group(P＞0.05). However, neuron counting was significantly higher in the EBHM+NMDA group than normal saline +NMDA group at 14days after intravitreal injection (P=0.044), but was lowered than normal control group (P＜0.05).· CONCLUSION: EBHM has no effect on neuron counting of RGCL when administered alone in normal rats.The results indicates that EBHM plays a partial protective role in NMDA-induced neuron loss in RGCL in the rats.

Objective To analyze the main clinical features of human infection with H7N9 avian influenza complicated by acute respiratory distress syndrome (ARDS).Methods A retrospective analysis of complete clinical data of 9 cases of human infection with H7N9 avian influenza complicated by ARDS admitted from March 2013 to December 2014 admitted to Department of Critical Care Medicine of the Affiliated Drum Tower Hospital of Nanjing University Medical School was conducted.Results Nine patients' mean age was (46.3±12.3) years, male accounting for 66.7% (6/9). The main clinical features: ① In the duration of acute phase, high fever, cough, hemoptysis sputum, reduction of white blood cell count (WBC) and platelet count (PLT), and myocardial enzyme elevation were the features of the disease. Chest CT showed pulmonary consolidation and ground-glass like shadows. ② On admission, all their oxygenation indexes (PaO2/FiO2) were less than 200 mmHg (1 mmHg = 0.133 kPa) of which 66.7% (6/9) was less than 100 mmHg, mean (98.9±62.8) mmHg; 55.6% (5/9) required invasive mechanical ventilatory support; 77.8% (7/9) combined with shock, and hemodynamic monitoring showed peripheral vascular resistance was decreased. ③ There were secondary bloodstream infection in 5 cases and lung infection in 4 cases, accounting for 77.8% (7/9). ④ In 22.2% (2/9) patients, the virus relapsed after the anti-virus therapy was stopped for 7 days, then immediately antiviral treatment was used again, it was still effective.Conclusion Human infection with H7N9 avian influenza complicated by ARDS has typical clinical symptoms, laboratory tests and imaging features, often associated with distributive shock, and at the late stage, secondary pulmonary or blood infection and the virus resurgence may occur.

Objective To observe the effects of Jianpi Huoxue Prescription on the subcutaneous transplantation of tumor cell apoptosis and angiogenesis; To discuss its possible mechanism of action. Methods SMMC-7721 hepatoma mice models were established. 60 tumor bearing male mice were randomly divided into blank group, model group, 5-Fu group, Jianpi Huoxue Prescription high-, medium- and low-dosage groups, with 10 rats in each group. Model group and Jianpi Huoxue Prescription high-, medium- and low-dosage groups were given medicine with relevant concentrations for gavage, once a day. 5-Fu group was given 5-Fu for intraperitoneal injection every other day. The weight, tumor weight and tumor inhibitory rate of each group were compared two weeks later. The protein expressions of VEGF and VEGFR-2 were detected by immunohistochemistry, microvessel density (MVD) were calculated, and the expressions of Survivin and Caspase-3 mRNA were detected by RT-qPCR. Results Compared with model group, protein expressions of VEGF and VEGFR-2, MVD, Caspase-3 and Survivin mRNA decreased in Jianpi Huoxue Prescription high-, medium- and low-dosage groups, and the most obvious differences were in 5-Fu group and Jianpi Huoxue Prescription high-dosage group (P<0.05). Conclusion Jianpi Huoxue Prescription can inhibit tumor growth by inhibiting angiogenesis and inducing apoptosis.

Adult ; Delivery of Health Care ; Female ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Occupational Health

The trace chemical components in functional Monascus rice were studied to explore the potential bioactive substances. MCI column, Sephadex LH-20 gel, and preparative liquid chromatography were used to purified the ethyl acetate extract from functional Monascus rice. Two novel pyridine Monascus pigments were isolated and identified, named monascopyridine G (1) and monascopyridine H (2), respectively based on extensive mass spectrometry (MS), infrared radiation (IR), and nuclear magnetic resonance (NMR) analysis. The molecular docking experiments between compounds 1 and 2 and peroxisome proliferators-activated receptor-gamma (PPARγ) showed that they exhibited obvious binding force with the receptor protein. Besides, the biosynthetic pathways of the two compounds were proposed, which provide a valuable reference for the selective production of these potential bioactive substances.

Liver Diseases ; Natural Killer T-Cells

Animals ; Fatty Liver ; etiology ; metabolism ; Humans ; Liver ; metabolism ; PPAR alpha ; physiology

OBJECTIVETo extract RNA from Pinellia ternata and lay a foundation for studying the formation mechanism of P. ternata.

METHODBy modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds, a simple and convenient method for extraction of total RNA from the tubers, stems and leaves of P. ternate containing abundant polyphenols and polysaccharides was established. High concentrated p-mercaptoethanol was added in the RNA extracted buffer to remove polyphenols, phenol and chloroform were used to eliminate proteins, and isopropanol and sodium acetate were used to precipitate polysaccharides.

RESULTThe A260/A230 value of RNA extracted with improved method were all over 2.0 and the values of A260/A280 were between 1.7 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good.

CONCLUSIONThe results showed that RNA obtained from the tubers, stems and leaves of P. ternate with this method had high purity and quality and could be used in molecular biological research, as DDRT-PCR and reverse Northern blotting analysis directly. This method is simple, economic, stable performance, and has a good repeatability as well as is suitable for extracting total RNA of medicinal plants with high concentrations of phenolics and polysaccharides.

Blotting, Northern ; Pinellia ; genetics ; Plant Leaves ; genetics ; Plant Stems ; genetics ; Plant Tubers ; genetics ; Polymerase Chain Reaction ; RNA, Plant ; isolation & purification

Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.