To construct a cDNA library of gerbil′s cochlea, the mRNA was separated from the cochlea of gerbil and the first strand cDNA was synthesized through reverse transcription by a modified oligo(dt) primer. The double strand cDNA was amplified by Ld pcR. After cDNA size fractionation , the ds cDNA was ligated in the ? TripIEx2 vector . The cDNA library was identified with special primers of prestin genes of cochlea by PCR .The results showed that the titer of library was 1.8?10 6 pfu and the percentage of recombinant clones was 80% . Prestin gene was contained in the library , the size of the fragment was 863bp . The results suggest that the established cDNA library has a high titer, recombinant percentage and large insert fragments of genes . The study lays the foundation of molecular biological study of the cochlea.

Objective To investigate the risk factors of aneurysmal subarachnoid hemorrhage (aneuryismal sub?arachnoid hemorrhage, aSAH) vasospasm (cerebral vasospasm, CVS) and provide the basis for the clinical prevention and treatment of CVS. Methods A retrospective analysis of clinical data was conducted on 255 cases aSAH patients receiving treatment between March 2012 and March 2014 in First Affiliated Hospital of Xinjiang Medical University Department of Neurosurgery, s treated. The clinical data included admission age, gender, ethnicity, history of hypertension, smoking his?tory, arterial tumor site, improved Fisher grading, admission Hunt-Hess grade, the dosage of Nimodipine, dehydrating agent, white blood cell count, blood glucose, blood lipids, blood calcium levels, platelet count. Univariate analysis and multivariate Logistic retrospective analysis were used to analyze the association between above-mentioned factors and the occurrence of CVS. Results A total of 73 cases developed CVS after aSAH and incidence rate of CVS was 28.6%. Uni?variate analysis showed that there were significant differences between patients with and without CVS in history of hyper? tension, smoking history, improved Fisher grade, admission Hunt-Hess grade, small doses of nimodipine, white blood cell count and blood glucose (P<0.05). The Logistic regression analysis showed that the history of hypertension, smoking history, improved Fisher grade, admission Hunt-Hess grade, a small dose of Nimodipine and white blood cell count were risk factors of CVS after aSAH (P<0.05). Conclusions the History of hypertension, smoking history, improved high Fish?er grade, high admission Hunt-Hess grade are independent risk factors of CVS after aSAH. A small dose of Nimodipine is a protective factor while increase in white blood cell count is a risk predictor, which should be controlled by enhancing clinical prevention.

Objective To explore the technique and effect of selected three-field lymphadenectomy by left thoracotomy in treatment of thoracic middle or lower section esophageal squamous carcinoma. Methods From Jun. 2005 to Mar. 2009, 213 patients with thoracic middle or lower section of esophageal carcinoma received esophagectomy, bilateral mediastinal lymphadenectomy and pleural membrane resection.Group 1 -5, 7 - 12a, 16al, and 19 were performed to dissect abdominal lymph node and extended thoracic and abdominal lymphadenectomy and only lymph node extraction of mesoesophagus in neck field. Results 14197 lymphatic nodes(LN) were detected in 213 case. The average number of resected LN was 66. 65 ±24. 73. The metastatic lymph node was detected in 105 cases. The metastatic rate was 49.05% (105/213).There were 423 metastatic lymph nodes. The lymph nodes metastasis was 2. 97% (423/14197) of all dissected lymphatic nodes. No remnant carcinoma in the upper and lower cutting edge was found in pathological examination. The operation time ranged from 2. 92 ～ 4. 67 ( 3. 37 ± 0. 42) hours. Blood transfusion during perioperative period was 0 ～ 6u ( 1.08 ± 0. 93 ) u. Perioperative plasma transfusion was 0 ～ 1400( 103.77 ± 184. 89) ml. The hospital-time was 14 ～ 39 ( 17.64 ±4. 12) days. There were no anastomotic leakage and recurrent laryngeal nerve injury. One case died from respiratory failure, the mortality was 0. 04% ( 1/213). Conclusion Surgical approach in the management of left thoracotomy in the sixth intercostals could extend resection of chest-field lymph node dissection, decrease neck field lymph node dissection. Abdomen-field lymph node dissection reached selected D3. The selected lymphadenectomy procedure had the advantages of small traumas and few complications.

INDENA SPA Company in Italy is a multi-national company that produces and sells plant extracts. Based on its own re- search advantages in the field of Ginkgo biloba preparation, the company protects its own products market effectively through building patent portfolio around the patents of its opponent. Based on the multi-angle analysis for patent portfolio of G. biloba preparation from the aspects of application time, legal status, technical development route, and patent portfolio layout, this article provides technical reference on research and development of G. biloba preparation, and the author suggest that Chinese applicants learn techniques and layout experiences of other patents fully to enhance the level of research and patent protection level.
Chemistry, Pharmaceutical ; economics ; legislation & jurisprudence ; Ginkgo biloba ; chemistry ; Italy ; Patents as Topic ; Plant Extracts ; chemistry ; economics ; isolation & purification

Objective To investigate the exact protocol eliciting the hippocampal CA1 long-term depression (LTD) of rats in vivo and the effect of amyloid β-protein (Aβ) on the LTD.Method By applying test stimulation to Schaffer collateral in hippocampal CA1 region in rats,recorded the in vivo field excitatory postsynaptic potentials (fEPSPs) ;further,observed the induction of LTD with different low frequency stimulation (LFS) and investigated the effect of Aβ25-35 on the LTD.Results Prolonged LFS (1,5 and 10 Hz) but not paired-pulse stimulus (PPS) effectively elicited the LTD in the hippocampal CA1 region,with significantly decreased amplitude of fEPSPs after LFS ; 1 Hz 900 pulses group induced a stronger LTD,being (63.7 ± 3.8) ％ at 120 min post-LFS,lower (P ＜ 0.05) than (75.1 ± 3.2) ％ in 600 pulses group ; different frequencies (1,5 and 10 Hz) of LFS with same pulses induced similar degree of LTD,the amplitude of fEPSPs were (63.7 ± 3.8) ％,(61.2 ± 3.6) ％ and (59.8 ± 3.9) ％ respectively,without significant differences between any two groups (P ＞ 0.05) ; after applying 12.5 nmol and 25 nmol Aβ25-35,the amplitude of fEPSPs decreased to (63.2 ± 3.8) ％ and (46.8 ± 3.9) ％,respectively,and lower and than that in control ((73.9 ± 3.0) ％,P ＜ 0.05).Conclusion Prolonged LFS effectively induced in vivo hippocampal LTD of rats,which provides an important electrophysiological protocol for the study of synaptic plasticity; Aβ25-35 injection dont affect the baseline synaptic transmission,but dose-dependently enhance the in vivo hippocampal LTD of rats,indicating that Aβ-induced LTD facilitation may be involved the early impairment of learning and memory in Alzheimer's disease.

Animals ; Female ; Gangliosides ; pharmacology ; Hippocampus ; enzymology ; physiopathology ; Lead ; toxicity ; Learning ; drug effects ; Male ; Memory ; drug effects ; Nitric Oxide Synthase ; biosynthesis ; Random Allocation ; Rats ; Rats, Sprague-Dawley

AIM: To study the changes of plasma atrial natriuretic peptide(ANP), endothelin-1 (ET-1) and serum cardiac troponin I(cTnI) levels and the effects of ANP and ET-1 on myocardium in crush injury rats. METHODS: Crush injury was produced in SD rats,the serum levels of cardiac enzyme and cardiac troponin I,plasma levels of ANP and ET-1 were studied by automated biochemical analyzer,automated chemiluminescence assay and radioimmunoassay. RESULTS: The plasma ET-1 level was much lower ,the levels of plasma ANP and serum cardiac enzyme and cTnI were much higher after crush injury than those of the control group( P

AIM: To explore the role of activation and apoptosis of splenic lymphocytes in the development of viral myocarditis (VMC). METHODS: Apotosis and MHC II antigen of splenic lymphocytes were detected in the VMC group (VMC, death, 8 cases) and control group (non-cardiac death, 4 cases) with TUNEL and immunohistochemistry methods.RESULTS: Compared with the control group, the increased expression of MHC II antigen and apoptosis were found in the splenic lymphocytes in the VMC group. CONCLUSION: These results suggest that the abnormality of the apoptosis/activation of splenic lymphocytes may be involved in the pathogenesis of VMC.

It has been well known that apoptosis induction and cell cycle arrest are typical biological effects observed in cancer cells after proteasome inhibition. TH2 is a new natural xanthone analogue isolated from the resin of Garcinia hurburyi tree. Here, the cell growth inhibition of TH2 on human hepatocellular carcinoma cell line (Bel-7402) was evaluated in vitro using SRB assay. The treatment of 10 ?mol/L TH2 reduced the surviving fraction from 86% (12 h) to 17.2% (48 h). To assess whether TH2 induce apoptosis, the appearance of sub-G1 peak, a specific fraction for apoptosis was detected by flow cytometry analysis. Progressive increase in the percentage of apoptotic population was observed in a dose-and time-dependent manner. Furthermore, a cleavage of poly (ADP-ribose) polymerase (PARP), a marker of early apoptosis, was observed clearly when the cells exposed to 10 ?mol/L of TH2 for 24 h by immunoblotting analysis. In vitro activities of 20 S proteasome purified from human erythrocytes on fluorogenic peptide substrates revealed that TH2 inhibited the trypsin-like, chymotrypsin-like and peptidylglutamyl peptide hydrolyzing activities in dose-dependent manner. Moreover, the turnover of tumor suppressor p53, a sign of deregulation of cell cycle progression and apoptosis induction by classical proteasome inhibitors, was disrupted in Bel-7402 cells. All these data indicate that TH2 had inhibitory effect on the proliferation of Bel-7402 cells and induction of apoptosis, which might be related to its inhibition of proteasome.

Objective To investigate the protective effects of 1-hydroxy-2, 3, 5-trimethoxyxanthone (QGS) on acute lung injury of mice induced by ip lipopolysaccharide (LPS). Methods Mice were pretreated with QGS for 7 d. Murine models of acute lung injury were duplicated by injection of LPS 20 mg/kg intraperitoneally. In 12 h, the lung weight index was observed and the NO level in the bronchoalveolar lavage fluid (BALF) was measured with kits. The lung was also assessd for the expression of I-?B, inducible nitric oxide synthase (iNOS), and cyclooxygenase-Ⅱ (COX-2) using Western blotting analysis. Lung pathological changes were also observed by HE in each group. Results The lung weight index of injury lung in mice induced by LPS was decreased in 500 mg/kg QGS group (P