ObjectiveTo establish the human periodontal ligament fibroblasts(HPDLFs)that express BMP2 and observe their biologicl characterization. MethodsA phagemid expression vector for BMP2 (pBK-B2) was transfected into HPDLFs by using LipofectAMINE. The BMP2 expression was determined by the immunohistochemical ABC method. The alkaline phosphatase (ALP) activity, osteocalcin (OC) production and capacity of mineralization were measured in the transfected cells. ResultsBMP2 protein was expressed in HPDLFs after gene transfection. The BMP2 gene transfected cells showed prominently elevated ALP activity, OC production and increase in mineralized nodules. ConclusionThe results indicate that BMP2 is expressed in HPDLFs and is involved in inducing differ- entiation of HPDLFs into osteoblast-like cells.

Forty cases of type Ⅱ diabetes mellitus were treated by puncturing point Neiguan (PC 6), and the effect on their cardiac vegetative nerve functions were observed at 20 min, 40 min and 60 min after acupuncture respectively. The findings showed that all heart rate variables improved remarkably at the three time periods after acupuncture treatment, with significant differences (P＜0.01); but there was no significant difference in the curative effects among the three time periods.

Objective To explore the behavior characteristics in children with benign epilepsy combined with centro-temporal spikes(BECTS).Methods Eighty-two children with BECTS aged 2.5-3.0(2.65?2.31)years old,51 male,31 female,who were free of mental retardation assessed with Gesell developmental schedules,untreated with antiepileptic drugs,and were investigated 15 days after the latest seizure.Eighty-two healthy children with sex and age matched to the cases,53 male,29 female,aged 2.5-3.0(2.6?0.4)years old.The behavior characteristics of infants in BECTS group and control group were assessed with CBCL,including 6 behavior factors which were sleep problem,social flinches,depression,physical aspect,attacking,act of sabotage and the infants-middle school student social ability scale.Results The total scores of behavior characteristics and the scores of depression,sleep problem,attacking and act of sabotage in BECTS group were all higher than those in control group,the differences were statistically significant.However,scores of social flinches,and physical aspect in BECTS group had no significant differences compared with those of control group.There were no significant difference of social adaptive component between the BECTS and control group.Conclusions Children with BECTS have behavior disorders to some extent,but their social adaptive capacity are the same as normal children.

ObjectiveTo investigate influence of sedation and analgesia on stress reaction of post-cardiac surgery in infants with congenital heart disease.MethodsForty children with congenital heart disease were randomly divided into 2 groups after cardiac surgery.The analgesia group was given 0.5-2.0 ?g/(kg?h) fentanyl intravenous infusion in 20 children undergoing cardiovascular surgery.The control group was given 5-8 mg/(kg?dose) lbuprofen orally.Midaiolam 0.01-0.20 mg/(kg?h) was used in 2 groups for sedation by intravenous infusion or 0.05-0.10 mg/(kg?dose)by intravenous push intermittently.The effects and adverse effects of sedation and analgesia were observed on 2,8,24,48 h after surgery in each group.The levels of cortisol,growth hormone,insulin and blood glucose were measured,respectively.ResultsThere were significant differences in Ramsay,Comfort value on 2,8,24 h(Pa

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

Objective To compare the effect of electro-acupuncture(EA) and warm-needle moxibustion(WNM) on the foot three-dimensional gait time-space parameters and surface electromyography of post-stroke patients with flaccid paralysis. Methods A total of 61 patients were randomly divided into EA group (N=30) and WNM group(N=31). Both groups were given post-stroke conventional treatment, and additionally EA group received EA while WNM group was given WNM. Before treatment and after 2 treatment courses, we detected the foot three-dimensional gait time-space parameters of pace race, stance phase, swing phase, bilateral stance phase, and step length, and surface electromyography parameters of root mean square(RMS), integrated electromyogram (iEMG) and co-contraction ratio(CR) of tibialis anterior muscle and gastrocnemius at the state of maximal isometric voluntary contraction(MIVC). Results(1) After treatment, the foot three-dimensional gait time-space parameters of the two groups were improved in various degrees (P<0.05 or P<0.01 compared with those before treatment) , and the improvement of EA group was superior to that of WNM group (P < 0.05). (2) After treatment, the RMS and iEMG of the gastrocnemius and tibialis anterior muscle as well as CR of dorsal extensor at MIVC state were improved in various degrees (P < 0.05 or P < 0.01 compared with those before treatment) , and the improvement of EA group was superior to that of WNM group(P < 0.05 or P < 0.01). Conclusion WNM exerts better effect on improving the parameters of three-dimensional gait time-space parameters and surface electromyography of post-stroke patients with flaccid paralysis than EA.

AlM:To evaluate the expression of transcriptional factor lslet-1 in retina in experimental retinal neovascularization induced by oxygen.
METHODS: The murine retinal neovascularization were induced by hyperoxia exposure. The morphological observation of retinal neovascularization was performed using angiography by fluorescein dextran injection under the fluorescence microscope, and the new blood vessels were quantified after 5d in room air (17-day-old) by counting the vascular epithelial cell nuclei protruding into viteous cavity using HE stain. Realtime PCR and Western blot were used to examine retinal lslet-1 level in postnatal 7,12, 14,17 and 26d respectively.
RESULTS: A lots of new blood vessels were demonstrated in the mouse retina in hyperoxic group by fluorescein angiography and histological method. Moreover, no significant difference was found in retinal lslet-1 level in postnatal 7d between hyperoxic group and control group, but was significantly higher in postnatal 12, 14 and 17d mice compared with control mice. However, mice at postnatal 26d, expression of lslet-1 in retina decreased to normal level.
CONCLUSlON: ln processing mouse model of retinal neovascularization, sustained hypoxia retinal tissue induce retinal neovascularization by increas the expression of transcription factor lslet-1.

Acclimatization ; Altitude ; Electroencephalography ; Humans

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

This study is to explore new lead compounds by inhibition of Pin1 for anticancer therapy using temperature sensitive mutants. As Pin1 is conserved from yeast to human, we established a high-throughput screening method for Pin1 inhibitors, which employed yeast assay. This method led to the identification of one potent hits, 8-11. In vitro, 8-11 inhibited purified Pin1 enzyme activity with IC50 of (10.40 +/- 1.68) micromol x L(-1), induced G1 phase arrest and apoptosis, showed inhibitory effects on a series of cancer cell proliferation, reduced Cyclin D1 expression, was defined as reciprocally matched for protein-ligand complex in virtual docking analysis and reduced cell migration ability. In vivo, we could observe reduction of tumor volume after treatment with 8-11 in xenograft mice compared with vehicle DMSO treatment. Altogether, these results provide for the first time the involvement of 8-11 in the anticancer activity against Pin1.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Drug Screening Assays, Antitumor ; methods ; G1 Phase ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; NIMA-Interacting Peptidylprolyl Isomerase ; Neoplasms ; pathology ; Peptidylprolyl Isomerase ; antagonists & inhibitors ; Temperature ; Xenograft Model Antitumor Assays ; Yeasts