Adult ; Barotrauma ; diagnosis ; surgery ; Hand Injuries ; diagnosis ; surgery ; Humans ; Male ; Middle Aged ; Paint

Cell Transformation, Neoplastic ; drug effects ; genetics ; Dimethyl Sulfoxide ; pharmacology ; Electroporation ; methods ; HL-60 Cells ; Humans ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection

OBJECTIVETo develop an HILIC method for determination of dencichine in Sanqi tablet and evaluate the quality of Sanqi tablet of different hatches from various manufactures in the market.

METHODThe chromatographic separation was conducted on a Thermo HILIC column (4.6 mm x 250 mm, 5 microm) kept at 25 degrees C with acetonitrile and 0.1% H3PO4 (60:40) as the mobile phase. The flow rate was set at 1 mL x min(-1) and the detection wavelength was set at 213 nm.

RESULTThe contents of dencichine in Sanqi tablet ranged from 1.60 to 4.31 mg x g(-1).

CONCLUSIONThe well established method was successfully applied to determine dencichine in Sanqi tablet. The results demonstrated that this method was simple, accurate and could be applied for quality control of Sanqi as well as its associated preparations.

Amino Acids, Diamino ; analysis ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Tablets

This review introduced the anti-tumor effects of non-steroid anti-inflammatory drugs (NSAIDs) and summarized their possible molecular mechanisms according to recent abroad literatures and our research results. Some evidence showed that the anti-tumor mechanisms of NSAIDs were different in various tumors.NSAIDs decreased the biosynthesis of PGE_2 and regulated the expressions of downstream correlated genes and proteins through restraining abnormal expression of COX-2 in certain neoplasms,which resulted in the inhibition of tumor angiogenesis and proliferation as well as induced apoptosis. But in other cancer cells, NSAIDs, as activators of peroxisome proliferator-activated receptor ? (PPAR?), induced COX-2 expression, promoted the biosynthesis of cyclopentenone prostaglandins (cyPGs). cyPGs further induced tumor cell apoptosis with PPAR? dependently or PPAR? independently. Since their special mechanisms of anti-proliferation and pro-apoptosis, NSAIDs revealed significant synergistic effects with other anti-tumor treatments.

To investigate the role of ion channels in the coupling responses of neutrophils to extracellular stimulus, it is necessary to study the membrane ion channel activities using patch-clamp technique. However, little has been known about the ion channel activities in neutrophils due to the difficulties in forming giga-seal with pipettes because of small diameter of neutrophils and the easily developed polarization. Some studies indicated that favorable results could be achieved through pretreatment at low temperature before electrophysiological recordings. But it remains unclear whether the pretreatment affects the membrane current and why the seal rate increases after low temperature pretreatment. The purpose of this study was to investigate the effects of 4 degrees C pretreatment on the membrane current and cell polarity in human neutrophils. In the experiments, human neutrophils were isolated from fresh peripheral blood of healthy volunteers and divided into two groups (room temperature group and 4 degrees C pretreatment group). Voltage-dependent K(+) (Kv) currents were recorded in whole-cell voltage-clamp mode and large-conductance Ca(2+)-activated K(+) (BK(Ca)) currents were recorded using inside-out patches. The results showed that 4 degrees C pretreatment significantly inhibited cell polarity (P<0.05), and it took more time for neutrophils to form a polarity-cycle [(534+/-32) s, n=20] compared with those at room temperature [(257+/-24) s, n=20]. Meanwhile, seal rate significantly increased in 4 degrees C pretreatment group (64%) compared with that in the room temperature group (27.5%). The seal rate and cell polarity rate during 0 approximately 1 min after 4 degrees C pretreatment were significantly different from those at room temperature, while no significant difference was found during 9 approximately 10 min between the two groups. Our results suggest that 4 degrees C pretreatment can inhibit cell polarity and increase seal rate, but has no effects on membrane currents. It is also suggested that 0 approximately 1 min after 4 degrees C pretreatment is a more suitable time for electrophysiological recording in neutrophils.
Cell Polarity ; Cold Temperature ; Humans ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Membrane Potentials ; Neutrophils ; physiology ; Potassium Channels, Voltage-Gated ; physiology

The way of "extracting-salting-chromatography" was used to purify the phycoerythrin and phycocyanin from Porphyra yezoensis in process scale-up.First,by comprehensive comparison of efficiency,the Sephadex G-25 was selected from four resins (Sephadex G-25、G-100、S-300 and CL-6B) as the best choice used in crude extract desalting of phycobiliprotein.Then the preparation process of phycobiliprotein was scaled-up with raw material(Porphyra yezoensis) increased from 1g to 20g,and finally to 400g.The results indicated that the yields of purified phycoerythrin and phycocyanin (absorption spectra purity above 3.2) increased during according to process scale-up,with 0.323% phycoerythrin and 0.148% phycocyanin obtained from 400g frozen Porphyra yezoensis blades respectively.It is no doubt that the process involved in the experiment is a potential way for large scale preparation of phycobiliproteins of high purity.

AIMTo validate the abundance of Interleukin 8 receptor beta (IL-8Rbeta) mRNA in single human neutrophil.

METHODSHuman neutrophils were isolated and purified from volunteers, total RNA was extracted and a regular RT-PCR aiming at IL-8Rbeta mRNA was performed to ascertain its expression profile in human neutrophils and optimize the reaction conditions for the following single-cell RT-PCR procedures. Subsequently, single neutrophil or the cellular content was harvested to conduct reverse transcription and two-round PCR with the same primer pairs used before. Serial dilution of single neutrophil cDNA pool was carried out at the same time with the exact two-round PCR followed. The specificity of this single-cell RT-PCR procedure was verified by the BamHI restriction endonuclease digestion on the final cDNA products.

RESULTSRegular RT-PCR indicated IL-8Rbeta mRNA expression in human neutrophils. While single-cell RT-PCR was sensitive enough to detect trace IL-8Rbeta mRNA as predicted cDNA product could be amplified from a 10 000 times diluted intracellular specimen from single neutrophil, which indicated an abundant expression of this mRNA in human neutrophil. Moreover, BamHI digestion on the final cDNA product clarified the specificity of this single-cell RT-PCR procedure.

CONCLUSIONThis simplified semi-quantitative single-cell RT-PCR procedure specifically confirmed that IL-8Rbeta mRNA was highly expressed in human neutrophil, which also provided the possibility of comparing mRNA abundance at single cell level.

Cells, Cultured ; Humans ; Neutrophils ; chemistry ; RNA, Messenger ; genetics ; Receptors, Interleukin-8B ; analysis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Single-Cell Analysis ; methods

This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Liliaceae ; chemistry ; Rhizome ; chemistry

AIM AND METHODSTo observe the effect of temperature on burst opening of voltage-dependent K+ channels(Kv) in hypothalamic neurons by cell-attached mode of patch-clamp technique.

RESULTSWith temperature raising, the number of burst opening increased, so did its average burst duration. B1 and B2 raised from 1.5 ms and 6.6 ms at 32 degrees C to 8.1 ms and 83.2 ms (P < 0.05) respectively while the open number of inter-burst, from 1-2 to 8 (P < 0.05) too. Instead of SB, CB displayed predominantly a kind of burst opening.

CONCLUSIONMore burst opening of Kv in hypothalamic neurons with temperature raising, this was benefited to the body temperature regulation of neurons on hypothalamus.

Animals ; Hypothalamus ; cytology ; Neurons ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; physiology ; Rats ; Rats, Sprague-Dawley ; Temperature

A modified protocol for quick and economic treatment of cultured human umbilical vein endothelial cells has been established, which result in high viability of the cells to allow better performance in patch-clamp studies. The electrophysiological properties of Ca (2+)-activated K(+) (KCa) channel of the cultured cells were investigated with a cell-attached configuration. With this modified treatment method, the cultured cells appear fusiform in shape under microscope, and KCa channel currents could be detected readily, suggesting their eligibility for patch-clamp studies.
Cells, Cultured ; Endothelial Cells ; cytology ; physiology ; Humans ; Membrane Potentials ; physiology ; Patch-Clamp Techniques ; methods ; Potassium Channels, Calcium-Activated ; physiology ; Reproducibility of Results