The opening of laboratories in universities is one of important parts in teaching reform and it is necessary for bringing up high-qualified students.Combined with the practical teaching,we have a primary discussion with the problems of the laborato- ry opening and put forward some suggestions and measures.

To investigate the mechanism of agarwood formation in Aquilaria sinensis induced by Lasiodiplodia theobromae, the fermentation liquor of L. theobromae was analyzed qualitatively and quantitatively by gas chromatography-mass spectrometry (GC-MS). JAs were detected in the fermentation liquor. The effect of the fermentation liquor on the abundance of sesquiterpenes in the callus of A. sinensis was analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). And the fermentation liquor stimulated alpha-guaiene, alpha-humulene and delta-guaiene biosynthesis in calli. It was inferred that L. theobromae produced JAs, which resulted in a significant increase of sesquiterpenes in A. sinensis.
Ascomycota ; physiology ; Fermentation ; Sesquiterpenes ; metabolism ; Thymelaeaceae ; metabolism ; microbiology

BACKGROUNDDiagnosis and treatment for respiratory symptoms (RSs) of gastroesophageal reflux disease (GERD) is more difficult than that for common esophageal symptoms. The goal of this study was to evaluate the efficacy and safety of radiofrequency (RF) treatment on RSs of GERD in a preliminary 12-month follow-up observation.

METHODSFrom April 2006 to October 2008, 505 GERD patients with mainly respiratory presentations such as wheezing, chronic cough or hoarseness, were treated by endoscopic RF. A questionnaire was completed before and after treatment, using a six-point scale ranging from 0 to 5 to assess symptom severity and frequency. The symptom score was the sum of frequency and severity.

RESULTSSymptom scores were significantly improved at the end of the follow-up period. The mean heartburn score decreased from 5.31 to 1.79. The mean regurgitation score decreased from 5.02 to 1.64; mean cough score decreased from 6.77 to 2.85; mean wheezing score decreased from 7.83 to 3.07; and mean hoarseness score decreased from 5.13 to 1.81 (P < 0.01). No major complications or deaths occurred. Minor complications included temporary post-procedural retrosternal unease or pain (n = 106; 21.0%), mild fever (n = 86; 17.0%), transient nausea/vomiting (n = 97; 19.2%), and transient dysphagia (n = 42; 9.3%). Thirty-five (6.9%) patients had recurrence of symptoms. Endoscopic RF treatment was repeated in six patients, and laparoscopic fundoplication was performed in seven.

CONCLUSIONEndoscopic RF is an effective and safe means to treat RSs in patients with GERD.

Adult ; Aged ; Cough ; surgery ; Esophagogastric Junction ; physiopathology ; radiation effects ; Esophagoscopy ; methods ; Female ; Gastroesophageal Reflux ; physiopathology ; surgery ; Heartburn ; surgery ; Hoarseness ; surgery ; Humans ; Male ; Middle Aged ; Radio Waves ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the marginal microleakage of porcelain-fused-to-metal crown using four different cements.

METHODSSixteen porcelain-fused-to-metal crowns were built and randomly divided into 4 group, luted onto standard prepared human forward molars using four different cements (glass ionomer cement, resin-modified glass ionomer cement, PanaviaF, Super-Bond C&B adhesive luting system). After temperature cycling test, all the crowns were then submerged in 2% fuchsin for 24 h. The marginal microleakage at tooth cement interfaces was observed using light stereomicroscopy and evaluated in classification index. The marginal microleakage grade of 4 groups were analyzed by SPSS 13.0.

RESULTSThe PanaviaF demonstrated the least marginal microleakage, Super-Bond C&B adhesive luting system, resin-modified glass ionomer cement showed an intermediate level of marginal microleakage, glass ionomer cement was associated with severe marginal microleakage (total, Chi2 = 157.60, P < 0.01; among the different groups, P<0.05).

CONCLUSIONAdhesive resin luting system which is the first selection in clinical is better than glass ionomer cement and is good at porcelain-fused-to-metal crown.

Boron Compounds ; Cementation ; Crowns ; Dental Cements ; Dental Leakage ; Dental Marginal Adaptation ; Dental Porcelain ; Glass Ionomer Cements ; Humans ; Metals ; Methacrylates ; Methylmethacrylates ; Resin Cements

BACKGROUNDThe role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.

METHODSTo detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.

RESULTSIn 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).

CONCLUSIONSDICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.

Blotting, Western ; DEAD-box RNA Helicases ; analysis ; genetics ; physiology ; Endoribonucleases ; analysis ; genetics ; physiology ; Epigenesis, Genetic ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Ribonuclease III ; Stomach Neoplasms ; chemistry ; etiology ; genetics

S1 nuclease (from Aspergillus oryzae) is a specific enzyme to degrade single stranded DNA or RNA molecules. It has been reported to be able to convert superhelical circular DNA molecules into open circle or linear forms under certain conditions, but this function has not been well explored. In order to use the action of S1 nuclease to linearize circular DNA and develop a novel way of cloning microcircular DNAs, the pUC19 was used to investigate the relationship between the linearization efficiency of S1 nuclease and the amount of enzyme used. By this way the optimal conditions for linearization of circular DNAs by S1 nuclease would be determined. 0.3u to 17u S1 nuclease per 100ng pUC19 DNA was added into a 25 microL system, respectively, to perform the reaction. The effectiveness of enzyme digestion was realized by electrophoresis in a 1.2% agarose gel. The results showed that along with the increase in enzyme amount from 0.3u to 17u a gradual decrease in the superhelical form, a gradual increase in the linear form and then in the circular form was obvious. The conversion from superhelical form to linear and circular form was directly related to the enzyme amount used. A higher proportion of linear DNA molecules was achieved by using 5 to 17u S1 nuclease per 100ng DNA. Besides, electrophoretic mobility of the S1 nuclease-linearized pUC19 was the same as that of the linear form produced by restriction enzyme digestion. According to the result of phiX174 digested by S1 nuclease it has been proposed that the enzyme cleaves first randomly on one site of one strand, thus converting the superhelical molecules into open circle form, and then on the same site of the complementary strand to produce the linear form. Therefore, the S1 nuclease-linearized DNA molecules are intact in the sense of their length and can be used for cloning. The plasmid-like DNA pC3 from cucumber mitochondria is a double stranded circular DNA molecule with about 550bp and the smallest known plasmid-like DNA in eukaryotic mitochondria. Many attempts have been made to linearize the molecule by using restriction enzymes but failed. Therefore, S1 nuclease was used to linearize pC3 based on the results obtained with pUC19. The linearized pC3 DNA molecules formed a very sharp band in a 2.5% agarose gel after electrophoresis. They were then recovered from the gel, added an "A" tail and ligated with T-vector. After transformation into E. coli JM109 cells, the positive clones were, screened by the blue-white selection. The insert was then cut using restriction enzymes EcoRI and Pst I. The result of electrophoresis shows that the electrophoretic mobility of the insert is just the same as that predicted. A 32 P-labled probe was synthesized using pC3 as the template and Southern blot analysis was carried out. The result shows that the inserted DNA is hybridized to the probe, which indicates that the cloned DNA fragment is from pC3. The sequence information of the insert shows that the plasmid-like DNA pC3 was 537bp in length. The nucleotide sequence was deposited in the GenBank (the accession number is AF522195).
Blotting, Southern ; Cloning, Molecular ; methods ; DNA, Circular ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Single-Strand Specific DNA and RNA Endonucleases ; genetics ; metabolism

Adenoma, Sweat Gland ; metabolism ; Diagnosis, Differential ; Humans ; Keratin-14 ; metabolism ; Keratin-17 ; metabolism ; Keratin-18 ; metabolism ; Keratin-7 ; metabolism ; Keratins ; metabolism ; Papilloma ; metabolism ; Sebaceous Gland Neoplasms ; metabolism ; Skin Neoplasms ; metabolism ; Sweat Gland Neoplasms ; metabolism

Bacillus are well known antibiotic producers. In this study,dozens of Bacillus strains from different sources were screened. Among them,a strain with strong antifungal activity was found. With 16S rDNA test and Biolog assay,this strain was identified to be Bacillus amyloliquefaciens. The fermentation conditions were optimized in small conical flasks. After ammonium sulfate salting out,dialysis,freezing vacuum dehydration,the crude protein extracts were obtained. The thermal stability,pH stability,protease stability,ion stability and antifungal spectrum of this protein were studied further. Scanning electronic microscope was also used to explore the antifungal mechanism.

Objective To study the association between the levels of scrum resistin, visfatin and insulin resistance as well as β-cell dysfunction in the first-degree relatives (FDR) of type 2 diabetes mellitus (T2DM), and to investigate the role of these adipocytokins in pathogenesis of T2DM. Methods Serum levels of resistin, visfatin as well as fasting true insulin (FTI), proinsulin (FPI) levels were measured in 71 patients with newly diagnosed T2DM. 55 subjects with IGT/IFG and 174 NGT from first-degree relatives of T2DM, and 114 subjects of NGT without T2DM family history served as control group (NC). Insulin resistance was assessed by the homeostasis model assessment (HOMA-IR) and β-cell function was evaluated by HOMA-β and fasting PI-to-TI ratio (FPI/TI). Lipid profile, liver function and kidney function were also tested. Anthropometrical parameters such as body mass index (BMI), waist circmference and blood pressure were also recorded and life style and food intake spectrum investigated. Results (1)There were no significant differences of serum resistin levels among the four groups (P ＞ 0.05 ). The serum resistin level was not correlated with HomA-IR, HomA-β and obesity markers (P＞0.05). (2) The serum visfatin levels of DM group, IGT/IFG and NGT group were lower than the NC group (P＜0.05). There were no significant difference among DM group, IGT/IFG group and NGT. The serum visfatin level was not correlated with HOMA-IR and obesity markers (P＞0.05) , but negatively correlated with fasting blood glucose, 2 h postprandial blood glucose and blood pressure (P＜0.05). Conclusion The adipokine profile in FDRs of T2DM had distinctively altered before the development of impaired glucose tolerance. Serum levels of visfatin, showed a favorable effect on glucose metabolism also had a significant decrease on serum levels in the early stage of T2DM.

italic>Schisandra chinensis is a traditional Chinese medicine with the functions of reinforcing deficiency, strengthening, and inducing astringency, appliable to treat the chronic cough and deficiency in breath, palpitation, and insomnia, etc. A hybrid mass spectrometry scanning strategy (high-definition data-independent/data-dependent acquisition, HDDIDDA), enabling the ion mobility separation and alternating data-independent acquisition/data-dependent acquisition, was established, which, in combination with in-house library-driven automatic peak annotation workflows facilitated by the UNIFI software, was utilized to systematically characterize the multi-classes of chemical components from S. chinensis. The use of an HSS T3 column (100 mm × 2.1 mm, 1.8 μm), 0.1% formic acid in H₂O-acetonitrile as the mobile phase running at the flow rate of 0.3 mL·min^-1, and column temperature at 35 ℃, could enable good separation of the S. chinensis components within 42 min. HDDIDDA scan in both the positive and negative ion modes was employed for data acquisition. Based on the automatic peak annotation, reference standards comparison, MS² data interpretation, and literature analysis, we were able to identify or tentatively characterize 105 compounds in the S. chinensis decoction, involving 56 terpenoids, 42 lignans, five glycosides, one organic acid, and one flavonoid. HDDIDDA scanning can improve the coverage of data acquisition and improve the accuracy of identification, while CCS prediction analysis provides the possibility to distinguish isomers by the ion mobility technology. The results provide reference for the intelligent material basis research of TCM.