In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells (MSCs) stimulated by electromagnetic field (EMF) with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes: Bmp1, Bmp7, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expressions of Bmp1, Bmp7 mRNA of the stimulated group were up-regulated (P<0.05) and those of Egf, Egfr were down-regulated (P<0.05). It was suggested that the gene expression profiles of osteogenesis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell adhesion etc.
Bone Marrow Cells/*cytology ; Cell Differentiation ; Down-Regulation ; Electromagnetic Fields ; Gene Expression Profiling ; Gene Expression Regulation ; Mesenchymal Stem Cells/*cytology ; Nucleic Acid Hybridization ; Oligonucleotide Probes/chemistry ; Osteogenesis/*genetics ; RNA, Complementary/metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To compare the timeliness, success and mortality rates between the modified carotid artery puncture method ( MCAPM) and standard suture method ( SSM) in the establishment of rat model of a middle cerebral ar?tery occlusion ( MCAO) . Methods Thirty?two male rats were randomly and equally assigned into MCAPM group and SSM group. The MCAO models were established by inserting a thread into the common carotid artery ( CCA) . 24 h after modeling, the rats of the two groups were evaluated with Bederson neurological scores, and the modeling success rate and mortality rate were analyzed. Results The suture insertion times, success rates and mortality rates of the MCAPM vs. SSM groups were (82?3 ±17?4) s versus (164?6 ± 22?0) s (P<0?01), 87?5% versus 68?75% (P>0?05), and 6?25% versus 18?75% (P>0?05). Conclusions MCAPM can be used to establish the rat model of MCAO due to its simplicity, mild wound and feasibility.

AIM:To compare the efficiency of fibrin glue to suture technique in pterygium surgery performed with limbal autograft under different methods of anesthesia.
METHODS: A prospective randomised clinical trial was carried out in 60 eyes of 55 patients operated for primary nasal pterygium, which were divided into two groups randomly: experimental group ( 30 eyes in 27 patients ) was under surface anesthesia ( oxybuprocaine ) and control group ( 30 eyes in 28 patients ) was under local anesthesia ( 20g/L lidocaine ). Autologous limbal graft taken from the superotemporal limbus was used to cover the sclera by a fibrin tissue adhesive after pterygium excision. Patients were followed up at least for 6mo. Time of operation, matching degree of graft and VAS score were mainly observed and recorded.
RESULTS: In experimental group the average surgery time was shorter (P=0. 008) and matching degree of graft ( 93%) was better than control group ( 83%) , the differences had statistical significance(P<0.05).
CONCLUSION: The surface anesthesia is enough when using fibrin glue for graft fixation in pterygium surgery, which will shorten surgery time and get better matching degree of graft.

Objective To evaluate the application of high-resolution ultrasonography in the diagnosis of nerve root lesion of cervical vertebra intervertebral foramen part. Methods Twenty-seven patients with root avulsion injury of brachial plexus intervertebral part and 8 patients with schwannoma of brachial plexus root were examined with high-resolution ultrasonography. Thirty healthy volunteers were involved for the sonographic imagings of normal brachial plexus intervertebral part. The ultrasound findings of the patients before operation were compared with surgical outcomes. Results The normal intervertebral part of brachial plexus were shown as hypoechoic tubular strucuture, which could be identified with cervical artery and transverse process used as landmarks for the intervertebral part of brachial plexus root. The detection ratios of normal intervertebral part for C5-C8 and outside part of braehial plexus were 100％. In 27 patients with root avulsion injury of intervertebral brachial plexus, the intervertebral nerve root were enlarged as hypoechoic area, with nerve dissected partly or disappearing, distal nerve parts outside the intervertebral foramen swelling, edema,or cerebrospinal fluid assembling as cystic masses. In 8 patients with schwannoma of brachial plexus root part, the spinal ganglia outside intervertebral foramen were shown as tumor-like hypoechoic mass, with blood supply shown inside. Conclusions High-resolution ultrasonography is of great value for early and convenient diagnosis of root avulsion injury and mass of brachial plexus intervertebral part, especially in community hospitals,which could provide important information for clinical therapy.

Objective To investigate the significance of 25 hydroxyvitamin D3 (25 (OH)D3) in patients with type 2 diabetes mellitus by comparing the differences of serum 25-hydroxyvitamin D levels between type 2 diabetes mellitus patients and healthy people.Methods One hundred and forty-seven patients with type 2 diabetes mellitus were selected as the T2DM group,and 135 healthy people were selected as the normal control (NC) group.The level of serum 25 (OH) D3 was detected by ELISA.Clinical data were recorded and biochemical indexes were also detected.Statistical analysis was performed.Results (1) Serum 25 (OH) D3 level in T2DM group was low,but there was no significant difference between two groups((23.10 ± 1.59) μg/L vs.(23.11 ± 1.65) μg/L; t =0.064,P =0.949),as well as between men and women in the 2 populations (T2DM men vs.women(23.22 ± 1.54) μg/L vs.(22.95 ± 1.65) μg/L; t =0.209,P ＞0.05; NC men vs women(23.03 ± 1.73) μg/L vs.(23.18 ± 1.59) μg/L; t =0.436,P ＞0.05) ; (2)The prevalence of vitamin D insufficient and deficiency in T2DM group was high,but it was not statistically significant between two groups (98.0％ vs.94.8 ％ and 2.0％ vs.5.2％ ; x2 =2.034,P =0.154) ; (3) Serum 25 (OH) D3 in T2DM group has a significant negative correlation with triglycerides (r =-0.218,P ＜ 0.01).Conclusion (1) Vitamin D level was low in both the healthy and T2DM populations in the southern region of China,the prevalence of vitamin D insufficient and deficiency was up to 90％.(2)Triglyceride is the only independent risk factor to T2DM population.

OBJECTIVETo evaluate the affection of crushing technology on quality. The dissolution of Pills of Six Herbs with Rehmunnia prepared by different crushing technology was determined by taking the dissolution of Paeonol as test marker.

METHODThe Pills was prepared with the fine powder which was crushed by normal crusher or super fine crusher. The rotatory-basket method was used, and the cumulative dissolution percentage was determined by UV.

RESULTStatistics indicated there was a significant difference in dissolution parameter (T50) between super fine crushing powder Pills and normal fine-crushing powder Pills (P < 0.01), and there was a difference in dissolution of different batches of Pills of Six Herbs with Rehmunnia prepared by the normal crush technique (P < 0.05).

CONCLUSIONThe determination of dissolution of Pills of Six Herbs with Rehmunnia is necessary. In order to improve the quality of drugs, we should adopt the technique of super fine crushing in the preparation procedure.

Acetophenones ; analysis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; Particle Size ; Plants, Medicinal ; chemistry ; Quality Control ; Rehmannia ; chemistry ; Solubility ; Technology, Pharmaceutical ; methods

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.
Enzyme Inhibitors ; pharmacology ; Free Radical Scavengers ; pharmacology ; High-Throughput Screening Assays ; Superoxides ; Uric Acid ; Xanthine ; Xanthine Oxidase ; antagonists & inhibitors

In order to improve the efficiency of drug screening on serotonin transporter (SERT) inhibitors, a high-throughput screening (HTS) model is established in RBL-2H3 cells. The RBL-2H3 cells are very similar to the serotonin genetic neuro, in modulation of post-receptor mechanisms and transduction pathway of SERT reactivated. Depending on a fluorescence substrate ASP+ used in detection method of inhibitor rates, it's convenient, quick, accurate and effective, not making the environmental biohazard compared with radioactive experiments. Furthermore, biological screening model combined with computer aided virtual screening technique describing high-throughput virtual screening (HTVS). Bayesian classification method and molecular fingerprint similarity were applied to virtual screening technique, for screening compounds in compound library. Some compounds have been found, and then validated further by biological screening model. Combination of HTS and HTVS improves the efficiency of screening SERT inhibitors.
Animals ; Bayes Theorem ; Cell Line ; Drug Evaluation, Preclinical ; High-Throughput Screening Assays ; Models, Biological ; Rats ; Serotonin Plasma Membrane Transport Proteins ; metabolism ; Serotonin Uptake Inhibitors ; pharmacology

Objective To study the influence of pulsed electromagnetic fields on the expression of type I collagen by bone marrow mesenchymal stem cells and it's mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro.The third passage cells were harvested and exposed to pulsed electromagnetic fields(PEMFs)at 15 Hz and 1 mT 8 h/d for 3 days.A semi-quantitative RT-PCR technique was used to measure the type I collagen mRNA expression;ELISA and immunohistochemitistry techniques were used to measure type I collagen expression.Inhibitors and promoters of the cAMP-dependent protein kinase A(cAMP-PKA)pathway were added.After the cAMP-PKA pathway had been inhibited or promoted,the effects of the PEMF on type I collagen expression were measured again using ELISA and immunohistoehemistry.Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type I collagen(P≤0.01)in comparison with the controls. The type I collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added,and raised when the promoter 8-Br-cAMP was added.Conclusion PEMFs at 15 Hz 1 mT can promote type I collagen expression of bone marrow mesenchymal stem cells.and the effect is correlated with the cAMP-PKA pathway.

Child ; Child, Preschool ; Female ; Gastrointestinal Hemorrhage ; diagnosis ; Gastrointestinal Tract ; diagnostic imaging ; Humans ; Infant ; Male ; Radionuclide Imaging ; Sodium Pertechnetate Tc 99m