Object\ To develop a convenient and practical method for the identification of Carapax Trionycis Methods\ Based on the sequence variations of 12S rRNA gene between Pelodiscus sinensis and other softshell turtles, a pair of allele specific primers was designed to distinguish P. sinensis from other species of Trionychidae. DNA were extracted and anplified and Carapax Trionycis could be identified accurately by polymerase chain reaction (PCR) using the primers Results\ Ten samples of turtle shell from different sources were indentified by the allele specific PCR with the primers The result indicated that three samples were substitutes of Carapax Trionycis, consilient with the result from DNA sequence analysis The mitochondrial 12S rRNA gene fragment of P. maculatus and a faked imitation had also been sequenced Conclusion\ The primers could be used as key components in Carapax Trionycis identification kit

0.05).Meanwhile,the percentage of elongation increased following each recasting(P

Objective To study prognosis and grading of somatosensory evoked potential(SEP)in long-term unconscious patients after severe traumatic brain injury(TBI).Methods Five prognostic factors including age,sex,injury mechanism,history of temporal craniotomy and SEP grading were selected and analyzed in 47 patients after severe TBI with a duration of unconsciousness longer than two weeks.The prognosis was judged by Glasgow Outcome Scale.Results Prognosis was closely associated with SEP grading(P=0.024).The accuracy of SEP in assessing the prognosis was 91.5%.About 95%-100% of patients with SEP at grade Ⅲ-Ⅲ ended up with severe disability,persistent vegetative state or death.However,43.75% of patients with SEP at grade Ⅰ had good prognoses.Conclusions The SEP grading can objectively and accurately evaluate patients' prognosis and demonstrate the brain function.

Twenty-fine samples of stomach and 25 samples of adenocarcinoma stomach were employed for this study. The former have been collected from the patients of duodenal ulcer. Specimens were taken from the large curvature and pylorus. In the adenocarcinoma of stomach, materials were taken from the cancer focus and the cancer margins, fixed in 10% formalin-GaCl_2 Solution, embedded in paraffin and cut at 5? in thickness. Then they were examined with defferent reactions of histochemistry; PAS, Alcian blue (AB), Toludine blue (TB) and Barnett's method. The surface of gastric mucosa obtained from patients of duodenal ulcer after subtotalgastrectomy was covered with a substance which consists of neutral and acid mucopolysaccharide and protein. The epithelial cells of the mucosa contains large amount of neutral muco-polysaccharide. The cells of the neck of gastric gland contain quite a bit of acid muco-polysaccharide, but the cells of the deep segments of the glands do not show any of it. The cells of adenocarcinoma of stomach contain a different deal material mixture which consists of acid muco-polusaccharide and protein. The surface of the cavity in the well-differenlialed cancer glands was covered by a positive substance in TB, AB method, which forms a thin layer of membrance. An evenly distributed substance in the cavity of carcinoma gland was shown with positive reactions of PAS, AB, TB and protein. The degenerated cells show a positive reactions of PAS and protein. The cells of low-differentiated cancer were surrounded with a superficial layer of the acid mucopolysaccharide. The interstitia in the invaded margines often showed clear, distinct positive PAS and TB reactions. It indicates that it cortains more chondroitin sulfuric acid and the hyaline acid. In the superficial layers of mucosa of the cancer tissue, the epithelial cellswere often to have less positive substance than that of the membrane in the parts of cancer margine, it means a lesser amount of neutral-polysaccharide. But the acid muco-polysaccharide in the surface of cancer and its metaplastic margin clearly increases in amount than the metaplastic area far away from the cancer focus. This report discussed the significance of the increased muco-polysaccharide in the tissue of the gastric cancer.

AIM　It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS　DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS　Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION　The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market.

Objective To explore the effect of fibroblast growth factor receptor(FGFR)inhibitor BGJ398 on the vasculogenic mimicry(VM)formation of glioma cells. Methods The phosphor-FGFR(pFGFR)was de-tected by Western blot,the expressions of MMP2 and MMP14 were detected by Western blot and immunocytochem-istry;the VM formation of U87MG and U251MG was tested by tube formation assay;subcutaneously implanted tu-mor model in nude mouse was established and tumor sections were CD34/PAS double-stained to detect the forma-tion of VM in vivo. Results Western blot showed that pFGFR in the experimental groups decreased significantly (P < 0.05);western blot and immunohistochemical staining showed that the expression of MMP2 and MMP14 in the experimental groups decreased significantly compared to the control group. In the tube formation assay ,the tube formation of U87MG and U251MG cells were restrained. In the subcutaneously implanted tumor model ,the VM number of the experimental group(13.85 ± 3.96)was significantly lower than that in the control group(26.40 ± 5.06,P < 0.05). Conclusions In vivo and in vitro experiments confirmed that BGJ398 can inhibit the activa-tion of FGFR,and inhibit the VM formation of glioma cells. These indicate FGFR signaling pathway is involved in the formation of VM.

BACKGROUND: Dopamine is closely associated with occurrence of epilepsy and transmission in central nerval system, and its various functions are determined by specific receptors.OBJECTIVE: To establish temporal epilepsy model so as to probe into the influences of SCH23390, the antagonist of dopamine D1 receptors and haloperidol, the antagonist of dopamine D2 receptors injected in substantia nigra on temporal epileptic seizure induced by kainic acid and on electroencephalic activityDESIGN: Randomized controlled verified experiment.SETTING: Neurology Medicine Institute of Zhujiang Hospital Affiliated to Southern Medical University.MATERIALS: The experiment was performed in General Military Neurology Medicine Institute of Zhujiang Hospital Affiliated to First Military University of Chinese PLA from August to December 2004, in which, 30SD adult male rats were employed, massed varied from 250 to 300 g.METHODS: ① 30 rats were randomized into physiological saline (control) group (6 rats), kainic acid (KA) group (6 rats) and experimental group (18 rats). The experimental group was divided into 3 subgroups, named the antagonist of dopamine D1 receptors, SCH23390 + kainic acid group (D1 +KA group), the antagonist of dopamine D2 receptors,haloperidol + kainic acid group (D2+KA group) and physiological saline + kainic acid group (PS + KA group), 6 rats in each. In the control, physi ological saline 2 μL was injected in the right cerebral ventricle unilaterally. In KA group, kainic acid 2 μL was injected in the right ventricle. In each of experimental group, SCH23390, the antagonist of dopamine D1 re ceptors, haloperidol, the antagonist of dopamine D2 receptors and physio logical saline 1 μL for each was injected in substantia nigra on the right side successively and simultaneously, kainic acid 2 μL was injected in the right ventricle. ② Observed items: alters of EEG on the 0.5th 1st, 2nd, 6th and 24th hours after medication in each experimental group (compared with EEG of non-epileptic behavior, appearance of sharp wave, spike wave,sharp (spike) slow comprehensive wave and multi-spike slow wave determines epileptic activity) and changes in animal behaviors (0 grade: normal; Ⅰ grade: wet dog-like trembling, paroxysmal facial spasm, like winking,beard moving, rhythmic chawing; Ⅱ grade: rhythmic nodding; Ⅲ grade:paroxysmal spasm of anterior limbs; Ⅳ grade: paroxysmal spasm of bilateral anterior limbs when standing; Ⅴ grade: falling down, loss of balance and convulsion of four limbs). Cerebral hippocampal neural cell apoptosis was observed and the rats were sacrificed on the 5' day of medication. Cerebral hippocampal section was prepared and determined after in situ end labeling staining.MAIN OUTCOME MEAUSRES: ① Changes in behavior in rats before and after epilepsy and electroencephalogram (EEG) alters. ② Results of cerebra hippocampal neural cell apoptosis.RESULTS: Thirty rats entered result analysis. ① Epilepsy seizure: In the control group, there was no epilepsy attacked. In KA group, all of rats ap pear seizure, which attacked 10 minutes after KA injected in brain ventricle, reached the peak in 1 hour and stopped in 3 to 6 hours. ② EEG record: In the control group, there was not epileptic activity manifestations,like sharp wave, spike wave, spike slow comprehensive wave, etc. In KA group, epileptic wave presented in 10 minutes after injection, the seizure developed to the peak in about 1 hour, the wave amplitude was decreased in 3 to 6 hours, presenting paroxysmal slow and spike slow waves and no epileptic wave appeared after 12 hours. ③ Neuronal apoptosis: In the control group, few neural cell apoptosis was visible in hippocampus after injection.In KA group, neural cell apoptosis was visible obviously in hippocampus in 5 days after injection (P =0.00). With SCH23390, the antagonist of dopamine D1 receptors, hippocampal cell apoptosis was not reduced remarkably (P ＞0.05) and with haloperidol, the antagonist of dopamine D2 receptors injected in substantia nigra, hippocampal cell apoptosis was aggravated (P =0.00).CONCLUSION: Injection of SCH23390, the antagonist of dopamine D1 receptors in substantia nigra cannot block kainic acid inducing epilepsy and epileptic electroencephalic activity is not weakened remarkably. Injection of haloperidol,the antagonist of dopamine D2 receptors enhances epileptic electroencephalic activity in kainic acid induced epilepsy and increases cell apoptosis remarkably in cerebral hippocampal CA3 area.It is to explain that it is dopamine D2 acceptor that is involved in regulation of temporal epilepsy in substantia nigra rather than D1 acceptor.

BACKGROUND: Bone marrow mesenchymal cells, multiple-potential non-hematopoiefic stem cells adhering to the wall in vitro culture, can be induced to proliferate and differentiate towards neurons and glia cells.OBJECTIVE: To investigate the growth state of cat bone marrow mesenchymal cells in vitro culture, as well as the capability to differentiate towards neural stem cells.DESIGN: A randomized sampling study.SETTING: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: This study was carried out at the Central Laboratory of the General Military Neurological Research Institute, Zhujiang Hospital, Southern Medical University between January and December 2002. Twenty healthy home-raised cats, aged 1.0 - 2.0 years and weighing 2. 5 - 4.0 kg, male and female in half, were provided by the Animal Center of the First Military Medical University of Chinese PLA.INTERVENTIONS: Bone marrows were randomly aspirated from the left or right hindlimbs in order to separate bone marrow mesenchymal cells, then the bone marrow mesenchymal cells single cell suspension was co-cultured with neural stem cell culture media in vitro so as to induce differentiation to neural stem cells with tretinoin. CK2 type inverted optical microscope(Olympus,Japan) was used to observe the growth of bone marrow mesenchymal cells in vitro culture, as well as 4, 12, 24, 48 hours of induction upon eliminating or not eliminating the wall-adhering cells. Bone marrow mesenchymal cells in stem cell stage were identified under Olympus optical microscope with modified immunohistochemical staining.MAIN OUTCOME MEASURES: The growth state and the immunocytochemical staining of living bone marrow mesenchymal cells exposed to experimental intervention were observed under the Olympus inverted optical microscope.RESULTS: Data from the 20 cats were analyzed without loss. Reversed microscopic observation revealed that cat bone marrow mesenchymal cells becrame larger when cultured in vitro, which were rich in plasmic granules with prominence projecting, adhering to the wall and forming cell clones. These cells were then successively cultured, and imnunohistochemical staining analysis suggested that the passaged bone marrow mesenchymal cells could express neural stem cells-specific antigen Nestin and differentiate towards glia-like cells and neuron-like cells.CONCLUSION: Cat bone marrow mesenchymal cells possess the characteristics of stem cells; they can be amplified into cell clones and induced to express the property of neural glia cells and neuron-like cells under proper condition.

Objective To investigate the regulation of swimming motility by H-NS in Vibrio parahaemolyticus(VP). Methods VP was inoculated into the semi-solid swimming agar plate containing 1% Oxoid tryptone, 2% NaCl, 0.5%Difco Noble Agar, and 0.1% arabinose followed by incubation at 37℃ for 4.5 h before the diameters of bacterial lawns were measured.Total RNAs were extracted from the wild-type (WT) strains and the hns null mutant (Δhns), and the quantitative real-time( RT)-PCR( qRT-PCR) was carried out to calculate the transcriptional variation of flaA between WT andΔhns strains.The entire promoter DNA region of flaA was amplified and cloned into the lacZ fusion vector pHRP309 containing a promoterless lacZ gene. The recombinant lacZ reporter plasmid was transformed into WT and Δhns, respectively, to measure the β-galactosidase activities in cellular extracts using the β-galactosidase enzyme assay system. Results and Conclusion The phenotype results showed that swimming motility of VP was enhanced by H-NS.The qRT-PCR and LacZ fusion results indicated that the transcription of flaA was positively regulated by H-NS.Collectively, H-NS promotes the swimming motility of VP, at least partly, by activating the transcription of flaA.

Objective To optimize the preparation of nanoparticles encapsulating antisense oligodeoxynucleotides in a-butyleyanoacrylate carrier (ASODN in NP) and investigate their stability. Methods ASODN in NP were prepared by interfacial polymerization of butyleyanoacrylate (BCA). The formulation and technology of the prepared NP was optimized by using orthogonal design based on the single-factor experiment. The morphology of NP was examined by transmission electron microscope; The size and size distribution of NP were determined by Malvern laser granularity equipment;The encapsulation efficiency and drug loading were determined by HPLC; The ability of protecting oligodeoxynucleotides from serum was investigated on a 20% polyacrylamide-7 Murea sequencing gel (PAGE). Results The nanoparticles in the optimal conditions were of regular spherical surface and discrete. The average size was 97.1 nm,the average encapsulation efficiency and drug loading of ASODN in NP were 96.7% and 10.1% respectively; The oligonucleotides were more efficiently protected from degradation by nucleases than by oligonucleotides adsorbed into nanospheres.Conclusion ASODN in NP has good stability,encapsulation efficiency,drug loading and great potential for ASODN delivery.