OBJECTIVETo explore the intervention of baicalin on signal transduction and activating transcription factor expression of ulcerative colitis (UC) patients.

METHODSRecruited were UC patients at Outpatient Department of Digestive Disease, Inpatient Department of Digestive Disease, Center for Digestive Endoscopy of College City Branch, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Southern Hospital affiliated to Southern Medical University from June 2010 to January 2011. They were assigned to the UC group (33 cases) and the diarrhea-predominant irritable bowel syndrome (IBS-D) group (30 cases). Another 30 healthy subjects were recruited as a healthy control group. Peripheral blood mononuclear cells (PBMCs) in vitro intervened by different concentrations baicalin were taken from UC patients. IL23R gene expressions in vitro intervened by different concentrations baicalin were detected using Q-PCR. Expressions of signal transducer and activator of transcription 4 (STAT4) , STAT6, phosphorylated-STAT4 (p-STAT4), and p-STAT6 were detected using Western blot. Serum levels of IFN-γ, IL-4, IL-6, and IL-10 were measured by ELISA. Effects of different concentrations baicalin on expressions of PBMCs, and levels of IFN-γ, IL-4, IL-10 of UC patients were also detected.

RESULTSCompared with the negative control group, 40 µmol baicalin obviously decreased IL23R gene expression of UC patients (P <0. 01). Compared with the healthy control group and the IBS-D group, p-STAT4/STAT4 ratios increased, p-STAT6/STAT6 ratios decreased, levels of IFN-γ, IL-4, IL-10 all increased in the US group (all P <0. 05). Compared with the negative control, 5 and 10 µmol baicalin groups, 20 and 40 moL baicalin obviously decreased p-STAT4/STAT4 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously increased p-STAT6/STAT6 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously lowered levels of IFN-γ and IL-4, and elevated IL-10 levels (all P <0. 05).

CONCLUSION40 µmoL baicalin could in vitro inhibit p-STAT4/STAT4 ratios, adjust p-STAT6/STAT6 ratios and related cytokines, thereby balancing the immunity and relieving inflammatory reactions of UC.

Activating Transcription Factors ; metabolism ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Blotting, Western ; Colitis, Ulcerative ; drug therapy ; metabolism ; Cytokines ; metabolism ; Flavonoids ; therapeutic use ; Humans ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; Leukocytes, Mononuclear ; Medicine, Chinese Traditional ; Phosphorylation ; STAT6 Transcription Factor ; metabolism ; Signal Transduction

Seven compounds were isolated from Onychium japonicum by macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified by NMR, MS and other spectroscopic methods as onychone A (1), quercetin (2), quercetin-3-O-α-L-rhamnoside (3), kaempferol-7-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnopyranoside (5), (-)-prunin (6), and norathyriol (7). Compound 1 is a novel macrocyclic flavonoid, and all the others are reported from this plant for the first time. In vitro cytotoxic activities of compounds 1-7 were evaluated by MTS testing with five cancer cell lines. Compound 7 exhibited weak cytotoxicity against tumor cell lines A549, SMMC-7721, and SW480.

A new class of dendrimer polylysine poly(ethylene glycol)-lipid was designed and synthesized. The cationic polymer liposomes were prepared by the lipid film-extrusion and post-insertion two methods with these dendrimer polylysine poly(ethylene glycol)-lipid and other lipids. The structural properties of obtained cationic polymer liposomes were studied by laser light scattering and fluorescence spectrometer. It was demonstrated that the nano sized liposomes with different density of surface cationic charges can be prepared by either lipid film-extrusion or post-insertion methods, but post-insertion process did not affect drug loading, did not influence drug loading capacity and did not induce liposomal morphology and particle size. The density of positive charge does not affect the size and distribution of different liposomes size and distribution. It was the better choice for manufacture because post-insertion method did not cause early release of drug and size changes. Cell binding experiments show that cationic polymer liposomes, especially dendrimer polymer liposomes had higher local charge density, and therefore have dramatic non specific cell targeting ability.
Animals ; Biological Transport ; Cations ; Cell Line ; Cricetinae ; Drug Delivery Systems ; Kinetics ; Lipids ; chemistry ; pharmacokinetics ; Liposomes ; chemical synthesis ; chemistry ; pharmacokinetics ; Molecular Structure ; Nanoparticles ; Particle Size ; Polyethylene Glycols ; chemistry ; pharmacokinetics ; Polymers ; chemistry ; pharmacokinetics

Poly (2-ethylacrylic acid) (PEAA) alkylamide derivatives were synthesized for constructing pH-sensitive liposomes by partially modification of carboxylic groups of PEAA with chemical reaction. These lipid derivatives of PEAA were synthesized by partially modification of carboxylic groups of PEAA with alkylamines. The acid-sensitive polymer associated liposomes were obtained by the method of polymer self-insertion in aqueous solutions through inserting hydrophobic lipid anchors of the polymer PEAA derivatives into the outer layer of vesicles. Factor effects on polymer insertion into liposomes were evaluated and the pH-sensitivity of the polymer associated liposomes was studied by calcein release assay. The PEAA-assoeiated-liposomes were prepared successfully by the methods of self-insertion. The PEAA-associated-liposomes are shown to be stable at neutral pH. (1) There was no correlate of anchor density of PEAA with length of the alkyl chain, but was positively correlated with the degree of PEAA modification. (2) Polymer insertion increased with initial ratio of polymer to lipid. (3) Unerting hydrophobic lipidr acidic conditions the associated polymer induces membrane disruption and fusion. (4) The PEAA-associated-liposomes shown pH-sensitive drug release property under acidic conditions. The anchored-poly (ethylacrylic acid) lipid derivatives can be useful in developing a potential pH sensitive drug delivery system.
Acrylates ; chemical synthesis ; pharmacokinetics ; Animals ; COS Cells ; Cercopithecus aethiops ; Drug Delivery Systems ; methods ; Fluoresceins ; metabolism ; Hydrogen-Ion Concentration ; Liposomes ; chemistry ; pharmacokinetics ; Particle Size ; Polyunsaturated Alkamides ; chemical synthesis ; pharmacokinetics

Objective To evaluate the pathophysiological mechanism of reduced seizure threshold following neonatal seizures by measuring the expression changes in N-methyl-D-aspartate (NMDA)receptor 1(NR1)and gamma-aminobutyric acid receptor A α1(GABAARα1)immunoreactivity which are the basic functional subunit in NMDA receptor or GABAA receptor family. Methods Seizures were induced by the inhalant flurothyl in 24 neonatal rats,starting from postnatal day 6.The 24 rats were subdivided into single seizure group (seizure lasted 30 min) and recurrent seizure group (seizures were induced for 30 min/d, totally 6 d). At day 7 and day 75 after the last seizure, brain homogenates were made. The expressions of NR1 and GABAARα1 proteins in hippocampus were examined by Westem blotting analysis. Results NR1 expression did not change significantly in single seizure group and recurrent seizure group at day 75, but was enhanced significantly in recurrent seizure group at day 7(P＜0.05).Meanwhile,polypeptide levels of GABAARα1 receptor subullit in the rat hippocanlpus decreased significantly in single seizure-treated rats at day 75 and recurrent seizure-treated rats at day 7 and 75 (P＜0.05). Conclusions Recurrent or single prolonged status epilepticus in neonatal rats might cause long-term modification on NR1 and GABAARα1 expressions in hippocampus at post-seizure 7 d or in adult, with excessive activation of excitatory neuronal circuits involving both reduction of GABAARα1 receptor and/or enhancement of NR1. This phenomenon might play a key role in long-term reduction of seizure threshold induced by early life status seizures.

OBJECTIVETo study the changes of platelet in May-Hegglin anomaly (MHA) and the molecular pathogenesis mechanism.

METHODSPeripheral blood was drawn from the MHA proband, her father and her uncle. Platelet count and morphology were examined by automatic blood cell counter and microscopy, respectively. The platelet membrane protein was examined by flow cytometry. Membrane antibodies were determined by ELISA. PCR was used to amplify the exons 25, 31 approximately 32, 38 and 40 of the MYH 9 gene in the MHA patient and her diseased father. Furthermore, PCR products were sequenced, a specific point mutation was identified and inclusions (Dohle's body) in the neutrophil was detected by indirect immunofluorescence technique.

RESULTSIt was proved that in MHA patients, platelet count was higher by cell counter than by microscope (P < 0.01). Giant platelet was 94% but platelet membrane proteins (CD41, CD61, CD42A, CD42b) were in normal range. Membrane antibodies was undetectable. An A5521G mutation (GAG-->AAG) in the exon 38 was found in the proband and her diseased father, resulting in a characteristic change of NMMHC-A1841 (Glutamic acid-->Arginine), which was not found in other members of the family and in normal controls. Spindle-like inclusions with fluorescence were clearly displayed in neutrophil cytoplasm.

CONCLUSIONThe molecular pathogenesis mechanism of May-Hegglin anomaly is the mutation in MYH 9 gene.

Adult ; Base Sequence ; Blood Platelets ; metabolism ; pathology ; DNA Mutational Analysis ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Granulocytes ; metabolism ; pathology ; Humans ; Inclusion Bodies ; metabolism ; pathology ; Male ; Molecular Motor Proteins ; genetics ; Mutation ; Myosin Heavy Chains ; genetics ; Pedigree ; Platelet Count ; Platelet Membrane Glycoproteins ; metabolism ; Thrombocytopenia ; blood ; genetics ; pathology

OBJECTIVETo identify the parental origin of methyl-CpG-binding protein 2 (MECP2) gene mutations in Chinese patients with Rett syndrome.

METHODSSingle nucleotide polymorphisms (SNPs) in intron 3 of the MECP2 gene were analyzed by PCR and sequencing in 115 patients with Rett syndrome. Then sequencing of the SNP region was performed for the fathers of the patients who had at least one SNP, to determine which allele was from the father. Then allele-specific PCR was performed and the products were sequenced to see whether the allele from father or mother harbored the mutation.

RESULTSSeventy-six of the 115 patients had at least one SNP. Three hot SNPs were found in these patients. They were: IVS3+22C >G, IVS3+266C >T and IVS3+683C>T. Among the 76 cases, 73 had a paternal origin of MECP2 mutations, and the other 3 had a maternal origin. There were multiple types of MECP2 mutation of the paternal origin, including 4 frame shift, 2 deletion and 67 point (56C >T, 6C >G, 2A >G, 2G >T and 1A >T) mutations. The mutation types of the 3 patients with maternal origin included 2 frame shift and 1 point (C >T) mutation.

CONCLUSIONIn Chinese RTT patients, the MECP2 mutations are mostly of paternal origin.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Fathers ; Female ; Humans ; Male ; Methyl-CpG-Binding Protein 2 ; genetics ; Mothers ; Mutation ; genetics ; Parents ; Polymorphism, Single Nucleotide ; Rett Syndrome ; genetics

Objective To describe a simple and efficient method to obtain large quantities of higllly purified Schwann cells(SCs).Methods SCs were isolated from the sciatic and brachial nerves of 3-to 5-day-old newborn SD rats by collagenase digestion.The isolated SCs were plated at the density of3x105/mL for primary culture,several rounds oftrypsin digestion were performed within 72 h to purify SCs. Results Compared with the purification using 1.25 g/L trypsin digestion in serial differential detachment procedures, Our protocol allowed easier and more efficient separation of the SCs from the fibroblasts.Immunocytochemical staining showed that the purity of the SCs exceeded 95%.Conclusion The purification protocol of the SCs we established can be easily carried out and yields well reproducible results to obtain large quantities ofhighly purified SCs for transplantation studies.

Objective To analyze the feasibility of local LINGO-1 polyclonal antibody administration for treatment of spinal cord injury in adult rats. Methods Twenty-four adult female SD rats were randomized into sham-operated group, rabbit IgG group and LINGO-1 antibody group. In the latter two groups, partial transaction of the T9 segment of the spinal cord was performed to completely sever the dorsal eorticospinal tract, followed immediately by administration of rabbit IgG and anti-LINGO polyclonal antibody via a mini-osmotic pump, respectively. At 3 and 28 days after the operation, the T8～10 segments of the spinal cord were harvested to prepare cryosections, and immunofluorescence staining was used to analyze the penetration of LINGO-1 polyclonal antibody into the spinal cord tissue and its specific binding to LINGO-1 molecules. Results In LINGO-1 antibody group, the presence of rabbit antibodies was detected at the injured sites of the spinal cord at 3 and 28 days after the operation. The mean immunofluorescence density was significantly lower in L1NGO-1 antibody group than in rabbit IgG group at 3 days after the operation (P<0.05). In rabbit IgG group, the mean immunofluorescence density for LINGO-1 in the crysections pre-treated with LINGO-1 polyclonal antibody was significantly lower than that in sections pre-treated with rabbit IgG(P<0.05). Conclusion Locally administered LINGO-1 polyclonal antibody can penetrate into the injured sites in the spinal cord in a wide time window and recognizes LINGO-1 molecule specifically, suggesting the feasibility of passive immunotherapy for spinal cord injury.

Objective To probe the technical processes of the suboccipital retrosigmoidal craniotomy with replacement of bone flap and deep muscles incision and suture and its clinical application. Methods The data of 40 patients undergoing suboccipital craniotomy to the cerehello-pontine angle lesions, deep muscles incision and suture in our hospital from May, 2003 to May, 2005 were analyzed retrospectively. Results All the procedures of craniotomy including removal of the bone flap, deep muscles incision and suture were accomplished safely with an average operation time of 40-60 min. Two patients appeared post-operative subcutaneous hydrops without CSF leakage and the majority can move their head freely sixth day after the operation. Conclusion Suboccipital retrosigmoidal craniotomy with replacement of bone flap, and deep muscles incision and suture, being a safe and feasible method, may decrease the happening of postoperative CSF leakage and encephalocele, and accelerate the recover of head movement.