The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Förster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy.
Animals ; Anti-Bacterial Agents ; chemistry ; metabolism ; pharmacology ; Binding Sites ; Cattle ; Humans ; Kinetics ; Lactoferrin ; chemistry ; metabolism ; Pefloxacin ; chemistry ; metabolism ; pharmacology ; Protein Binding ; Protein Conformation ; Serum Albumin ; chemistry ; metabolism ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

OBJECTIVE To study the in virto interaction of allicin combined with cefoperazone against clinical isolates of Pseudomonas aeruginosa.METHODS The protocol was designed by checkerboard method and the MICs of allicin combined with cefoperazone against the 17 strains of sensitive and 14 strains of drug-resistant P.aeruginosa were determined by broth dilution method,the FIC index was calculated according to MIC results.The combined effects were confirmed by FIC index.RESULTS The percentage of the FIC index less than 0.5,from 0.5 to 1,from 1 to 2,and more than 2 was 41.2-64.3% 35.7-41.2% 0-17.6%,and 0%,respectively.CONCLUSIONS Synergism and additivity of allicin combined with cefoperazone against P.aeruginosa are their main action,there are little autonomy and no antagonism.Allicin can significantly improve the antibacterial activity of cefoperazone against drug-resistant P.aeruginosa.

Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of Ca2+, Mg2+, K+, and HCO3 - in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.
ATP-Binding Cassette Transporters/*chemistry/*genetics/metabolism ; Adenosine Triphosphate/metabolism ; Amino Acid Sequence ; Animals ; Calcium/metabolism ; *Cloning, Molecular ; Cryptosporidiosis/parasitology ; Cryptosporidium/chemistry/genetics/*metabolism ; Humans ; Iron/metabolism ; Mice ; Molecular Sequence Data ; Protein Structure, Tertiary ; Protozoan Proteins/*chemistry/*genetics/metabolism ; Sequence Alignment

To address the throughput limitations of digital nucleic acid analysis,a tricolor combination-based droplet coding technique was developed to achieve multiplex digital nucleic acid analysis with flexible throughput expansibility.To improve the analysis efficiency,a machine learning-based method was further developed for automatic decoding of color-coded droplet array.The machine learning algorithm empowered the computer program to automatically extract the color-position-quantity information of the droplets.By correlating this color-position-quantity of droplets before and after nucleic acid amplification,the proportion of positive droplets for each target was rapidly determined.This droplet decoding strategy was applied to multiplex digital nucleic acid analysis.The experimental results demonstrated that this droplet decoding method was fast and accurate,with a decoding process completed within 2 min.Furthermore,the droplet identification accuracy exceeded 99%.Additionally,the obtained nucleic acid quantification results exhibited a good correlation(R2＞0.99)with those reported by a commercial digital PCR instrument.

OBJECTIVETo investigate the effect of bortezomib alone or combined with harringtonine (HT) or arsenic trioxide (As2O3) on the proliferation capacity and apoptosis of HL-60/ADM cell line and fresh cells from refractory/relapse acute leukemia patients.

METHODSHL-60/ADM cells or refractory/relapse leukemia cells were incubated with bortezomib at different doses alone and in combination with HT or As2O3. The proliferation capacity was observed by MTT assay, cell apoptosis by fluorescence microscopy and flow cytometry. Intracellular concentration of daunorubicin (DNR) was determined by flow cytometry.

RESULTSIn bortezomib-treated HL-60/ADM cells, the proliferation inhibition rate and apoptotic cells increased in a time- and dose-dependent manner. 40 nmol/L bortezomib could maximally inhibit the proliferation of HL-60/ADM cells at 48 hours. 15 micromol/L As2O3 or 752 nmol/L HT combined with different doses of bortezomib could inhibit proliferation and induce apoptosis of HL-60/ADM cells. The As2O3 plus bortezomib or HT plus bortezomib showed a greater anticancer efficacy than either of the drugs alone (P < 0.05, P < 0.01). Bortezomib (10 nmol/L) could markedly enhance the intracellular accumulation of DNR in HL-60/ADM cells (P < 0.05).

CONCLUSIONSBortezomib can inhibit proliferation and induce apoptosis of HL-60/ADM cells and fresh refractory/relapse acute leukemia cells, especially combined with HT or As2O3.

Adolescent ; Adult ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Child ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; HL-60 Cells ; Harringtonines ; pharmacology ; Humans ; Male ; Oxides ; pharmacology ; Pyrazines ; pharmacology ; Young Adult

OBJECTIVETo observe the effect of Kangxin Capsule (KXC) on the expression of nerve growth factor (NGF) as well as the morphology and amount of nerve synapse in the cortical parietal lobe and hippocampus CA, area of vascular dementia ( VD) model rats.

METHODSThe model rats of VD made by photochemical reaction technique were randomly divided into five groups: the model group (MG), the high-dose, middle-dose and low-dose KXC groups (HDG, MDG and LDG), and the Western medicine hydergin control group (WMG). They were treated respectively with distilled water, high, middle and low dosage of KXC suspended liquid, and hydergin for a month. Besides, a blank group consisting of normal (non-model) rats was set up for control (CG). The ultrastructure of nerve synapse in the cortical parietal lobe and hippocampus CA1 area of the rats were observed and its density estimated. The condition of NGF positive neurons in the above-mentioned two regions were also observed by immunohistochemical stain.

RESULTSAll the KXC or hydergin treated groups demonstrated a normal amount of nerve synapse with integral structure in the cortical parietal lobe and hippocampus CA, area, which approached that in the CG and was superior to that in the MG. Also, the NGF positive neuron in all the treated groups was much more than that in MG with significant difference ( P<0.01), approaching to that in the CG.

CONCLUSIONKXC could elevate the expression of NGF in the cortical parietal lobe and hippocampus CA, area, preserve the number and morphology of synapse, thus to protect the function of nerve system from ischemic injury.

Animals ; Capsules ; Dementia, Vascular ; drug therapy ; metabolism ; pathology ; Disease Models, Animal ; Female ; Hippocampus ; chemistry ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Microscopy, Electron ; Nerve Growth Factors ; analysis ; Neurons ; ultrastructure ; Parietal Lobe ; chemistry ; Rats ; Rats, Wistar ; Synapses ; ultrastructure

OBJECTIVETo assess the antitumor efficacy and adverse effects of bortezomib either used alone or in combination with arsenic trioxide for transplanted tumor in nude mice.

METHODSNude mice bearing HL-60 cell xenografts were randomized into 4 groups to receive treatment with normal saline, bortezomib, arsenic trioxide, bortezomib plus arsenic trioxide. The tumor growth inhibition and general condition of the nude mice were observed, and in situ TUNEL assay and immunohistochemistry were performed on the transplanted tumors.

RESULTSBortezomib alone and in combination with arsenic trioxide could both inhibit the growth of the transplanted tumors, prolong the survival of the nude mice, and induce cell apoptosis and growth inhibition of the HL-60 cells in vivo, and the combined administration exhibited even better effects. The administration was well tolerated with causing manifest vital organ damages in the mice.

CONCLUSIONBortezomib in combination with arsenic trioxide has significant antitumor effect in nude mice bearing HL-60 cell xenografts possibly by inducing HL-60 cell apoptosis and growth inhibition without producing no significant adverse effects.

Animals ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Proliferation ; Disease Models, Animal ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; physiopathology ; Male ; Mice ; Mice, Nude ; Oxides ; pharmacology ; Pyrazines ; pharmacology ; Random Allocation ; Xenograft Model Antitumor Assays

A portable ECG monitor is introduced in the paper, which has a temporary intravenous and transesophageal fixable rate pacing function. During the PSVT attack, the tachyarrythmia can be stopped by the transesophageal cardiac pacing while the ECG signals are monitored. The instrument has some advantages such as small size, low price and good practicality. It is worth while introducing and popularizing it to all hospitals.
Electrocardiography, Ambulatory ; instrumentation ; Equipment Design ; Heart Block ; diagnosis ; therapy ; Heart Rate ; Humans ; Monitoring, Ambulatory ; instrumentation ; Pacemaker, Artificial ; classification ; Signal Processing, Computer-Assisted ; Software ; Tachycardia ; diagnosis ; therapy

BACKGROUND: Although Becker's nevus (BN) is a relatively common disease, the systematic studies of clinicopathological and immunohistochemical results are poorly reported. OBJECTIVE: To investigate the clinicopathological features and immunohistochemical alterations of keratinocyte proliferation, melanocyte density, smooth muscle hyperplasia and nerve fiber distribution in BN. METHODS: Clinical and pathological data were collected in 60 newly-diagnosed BN cases. Immunohistochemical stain of Ki-67, Melan-A, keratin 15, smooth muscle actin and protein gene product 9.5 was performed in 21 cases. RESULTS: The median diagnostic and onset age was 17 and 12 years, respectively. Skin lesions usually appeared on the upper trunk and upper limbs. The pathological features included the rete ridge elongation and fusion and basal hyperpigmentation. Epidermal Ki-67, Melan-A and keratin 15 expression and dermal nerve fiber length were significantly higher in lesional and perilesional skin than in normal skin (p<0.05~0.01), while smooth muscle actin expression was upregulated only in skin lesion (p<0.05). CONCLUSION: Although the clinical diagnosis of BN is often straightforward, histopathology is helpful to differentiate from other pigmentary disorders. The hyperproliferation of keratinocytes, melanocytes, arrector pili muscle and dermal nerve fibers could be involved in the pathogenesis of BN.
Actins ; Age of Onset ; Diagnosis ; Hyperpigmentation ; Hyperplasia* ; Keratin-15 ; Keratinocytes* ; MART-1 Antigen ; Melanocytes* ; Muscle, Smooth* ; Nerve Fibers* ; Nevus* ; Skin ; Upper Extremity