Objective To analyze and compare the Eucommia ulmoides leaves from different producing areas by IR;To establish a TLC method for identification of Eucommia ulmoides leaves;To provide a reference for its quality control. Methods The characteristic absorption peaks were identified and the positions and intensities of Eucommia ulmoides leaves from 6 producing areas were compared by IR and second derivative spectrum, including clustering analysis; Separation and identification of the active components in Eucommia ulmoides leaves were conducted by TLC. Results There a difference exited in the characteristic absorption peaks in peakshapes, positions and intensities. And the samples from 6 producing areas could be divided into one category. Results found 6 batches of sample spots of consistent by TLC, but between samples of different producing areas trace components spots existed differences. Conclusion The two methods are direct, simple, fast and convenient, which can provide a reliable evidence for identification and quality control of the Eucommia ulmoides leaves.

Aim: To establish the HPLC and LC-MC methods for the quality analysis of the different species of Radix Paeoniae Alba cultivated in Bozhou. Methods: The LC-MS method was applied to analyze the chemical composition of the cultivated species, the HPLC method was applied to determine the contents of paeoniforin, ben-zoic acid and paeonivayin in crude drugs and processing products. Results: 10 characteristic peaks were identified by LC-MS with reference standards. The significant difference was observed in the chemical compositions of 3 cultivated species. The significant difference was observed in the chemical compositions of 3 cultivated species. The content of paeoniflorin was increased significantly after traditional processing. The results indicated the 4-year-old pubang and 5-year-old xiantiao are the best cultivated species. Conclusion: The methods are simple, rapid and precise to assess and control the quality of different species of Radix Paeoniae Alba cultivated in Bozhou.

OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.

[ ABSTRACT] AIM: To investigate the role of anterior thalamic nucleus in trigeminal nerve electrostimulation ( TNS)-induced effects on seizures and hippocampal damage.METHODS: The rats were intraperitoneally injected with pilocarpine to induce chronic epilepsy, and then received sham treatment, TNS treatment and TNS treatment after stereotac-tic lesion to the anterior thalamic nucleus.The TNS treatment lasted for 1 month in each group.Another injection of pilo-carpine was conducted to induce seizures, and the severity and duration of seizures were quantitatively evaluated.TUNEL, Fluoro-Jade B ( FJB) and Nissl staining were applied to determine in situ apoptosis, neuronal degeneration and neuron loss in the hippocampal CA1 area respectively.RESULTS:Compared with TNS group, TNS treatment after stereotactic lesion to the anterior thalamic nucleus significantly increased the severity and duration of seizures (P<0.05), and the numbers of TUNEL positive cells, FJB positive cells and lost neurons in the hippocampal CA1 area (P<0.01).CONCLUSION:Anterior thalamic nucleus plays a role in TNS-induced reduction of seizures and hippocampal damage.The mechanisms might be due to the chronically activation of the cortex through anterior thalamic nucleus pathway induced by TNS, resulting in the down-regulation of neuronal excitatory susceptibility.

Objective: To explore the role of angiotensin receptor type I (AT1)-calcineurin (CaN) signaling pathway in transient outward potassium ion channel (Ito) remolding in hypertrophic atrial myocytes of neonatal rats. Methods: 1 day old neonatal rats’ atrial myocytes were isolated and the cells were divided into 4 groups:①Control group, normal cells were cultured for 24 h,②Stretching group, the cells were cultured for 24 h with mechanical stretching to induce hypertrophy,③Telmisartan group, the cells were treated by telmisartan at 1 μmol/L for 1 h, then cultured for 24 h and ④Cyclosporin-A (CsA) group, the cells were treated by CsA at 0.25 μg/ml for 1 h, then cultured for 24 h. The ratios of protein/DNA in myocytes were compared between Control group and Stretching group, cell hypertrophy was deifned by mRNA expression of atrial natriuretic peptide (ANP). Ito changes were detected by whole-cell patch clamping technique, proteins expressions of Kv4.3 and CaN A subunit were examined by Western blot analysis. Results: Compared with Control group, Stretching group showed obviously decreased Ito density and Kv4.3 protein expression, while increased CaN A protein expression; Compared with Stretching group, the above effects were reduced in Telmisartan group and CsA group. Conclusion: AT1-CaN signaling pathway was involved in the regulation of Ito channel remodeling in hypertrophic atrial myocytes of neonatal rats.

Objective To explore the diagnostic significance of Xpert MTB/RIF in cerebrospinal fluid,and evaluate the application for early diagnosis of tuberculous meningitis(TBM).Methods Sixty cases of TBM and 30 cases of non-TBM patients were selected as our subjects.Xpert MTB/RIF and modified Ziehl-Neelsen stain were performed in cerebrospinal fluid.The detection rate of the system and the resistance of the patients were analyzed.Results Eleven cases were diagnosed as the positive cases in 60 cases with TBM,and 0 case was diagnosed as TBM in control group.Sensitivity and specificity of Xpert with TBM were 18.33％ and 100％,respectively.The difference of the two groups was statistically significant (P =0.014).The positive rate of definite group was 23.68％(9 cases),18.18％(2 cases) in probable group and 0％ in possible group,and the difference of the three groups was statistically significant(x2 =3.070,P＞0.05).The resistance rate was 36.36％ (4/11).Sensitivity of the modified Ziehl-Neelsen staín was 63.33％ (38/60).Eleven cases were detected positive by Xpert MTB/RIF,9 cases were positive with modified acid fast staining,and the positive rate was 18.33％,and the difference of the two methods was statistically significant (P =0.000).Conclusion Xpert MTB/RIF test is simple and rapid diagnostic method.The combination of Xpert MTB/RIF and modified ZiehlNeelsen stain will improve the efficiency of the early diagnosis of TBM.

Objective Toevaluatea modified Ziehl-Neelsen(Z-N) stain in the diagnosis of tuberculous meningitis. Methods Cerebrospinal fluid specimens from 35 patients were stained by using the modified Ziehl-Neelsen staining. Re-sults The positive rate was 94.29% in 35 patients with tuberculous meningitisand the intracellular acid-fast bacilli was detected in 53.40%of all specimens. One case was stained positive in 15 patients with non-tuberculous meningitis. Con-clusion The modified Ziehl-Neelsen stain not only significantly improves the detection rates of tuberculous meningitisbut alsois able to identify intracellular M.tuberculosisin cerebrospinal fluidspecimen.Thus, the modified Z-N stain can be a convenient tool for diagnosing tuberculous meningitis.

Objective To explore the best treatment strategy of pyogenic brain abscesses.Methods Clinical data of 45 patients(34 males and 11 females,ages ranged from 7 to 76 years,averaged 42.6 years) with brain abscess treated from January 1999 to March 2008 were retrospectively analyzed.Among the 45 patients,there were 40 with single brain abscess and 5 with multiple brain abscess;43 with unilocular abscess and 2 with multilocular abscess.19 patients accepted the conservative treatment with the third generation cephalosporin,16 were treated with stereotactic puncture of abscess,8 treated with stereotactic puncture and drainage,and microsurgical excision was performed in 5 patients.Results Of the 19 patients treated with conservative method,16 were cured,no recurrence was found during the follow-up period(from 6 months to 5 years) in 13 patients,and another 3 were deteriorated and accepted stereotactic operation.All the stereotactic operations were carried successfully out in 24 patients(including 3 cases after expectant treatment failure) and only one patient needed another stereotactic aspiration two weeks later because of abscess recurrence after the initial operation.Epileptic seizure was found in one patient at the end of stereotactic procedure.Total removal of brain abscess was performed by microsurgical craniotomy in 5 patients.At the time of discharge,clinical symptoms disappeared or were improved remarkably,and CT or MRI re-examination disclosed the disappearance or obviously diminution of abscess in all patients.No abscess recurred during the follow-up period(from 4 months to 3 years,average 14 months) in 22 patients.Conclusion A set of treatment strategy of pyogenic brain abscesses has been proposed and stereotactic operation seems to be the most appropriate surgical choice.

ObjectiveTo investigate the clinical value of endoscopic esophageal submucosal tunnel resection of gastric fundus-cardiac tumors originating from muscularis propria.Methods Clinical date of 18 patients with gastric fundus-cardiac submucosal tumors originating from muscularis propria who underwent endoscopic esophageal submucosal tunnel resection from January 2011 to December 2011 were retrospectively collected and analyzed.ResultsAll lesions were successfully and completely resected in 18 patients,with sizes ranging from 0.7 cm to 7.2 cm,mean (2.43 ± 1.91 ) cm.Pneumoretroperitoneum,pneumomediastinum and pneumohypoderma occured during the procedure in 2 cases,but spontaneously resolved in 3 days.Fever with increased WBC within 24 h after the procedure occurred in one patient,and was cured in two days with antibiotics.There were no severe complications including bleeding,perforation or death.All patients could have liquid diet 3 days later after the operation.Follow-up endoscopy at 1 week after the operation showed a healing of esophageal incision.ConclusionEndoscopic esophageal submucosal tunnel resection is a safe and effective method for gastric fundus-cardiac submucosal tumors originating from the muscularis propria,lessening the difficulty of traditional endoscopic resection.

Objective To investigate the effects of Saccharomyces boulardii ( S. boulardii) on the col-onization of Helicobacter suis ( H. suis) in stomach and the formation of gastric mucosa associated lymphoid tissue (MALT) during H. suis infection. Methods Sixty C57BL/6 wild type mice were randomly divided into six groups. The mice in group A and group B were respectively given sterile distilled water and S. boulardii twice by gavage and then infected with H. suis for one week. The mice in group C and group E were given sterile phos-phate buffer saline by gavage for one week and then respectively given sterile distilled water and S. boulardii by gavage twice a week for 12 weeks. The mice in group D and group F were infected with H. suis for one week and then respectively given sterile distilled water and S. boulardii by gavage twice a week for 12 weeks. Serum and gastric tissue samples were collected from each mouse. Results The bacterial loads of H. suis in the stomachs of mice in group B were significantly lower than those in group A. No significant differences in the levels of se-creted IgA( sIgA) in serum and gastric tissue samples and the expression of IFN-γat mRNA level in gastric mu-cosa samples were found between the two groups. The expression of H. suis 16S RNA and the formation of gastric lymphoid follicles were detected in mice in groups D and F. The levels of sIgA in serum and gastric tissue sam-ples and the expression of IFN-γ and CXCL13 at mRNA level in gastric mucosa samples increased significantly in groups D and F as compared with groups C and E. Compared with the mice in group D, the bacterial loads of H. suis in stomachs, the numbers of MALT per unit length of gastric mucosa and the expression of IFN-γ and CXCL13 at mRNA level in gastric mucosa decreased significantly in mice from group F, but the levels of sIgA in serum and gastric tissue samples increased significantly. Conclusion S. boulardii could inhibit the colonization of H. suis in stomach and suppress the formation of gastric MALT during H. suis infection.