The TSR1(thermosensitive rescued)gene associated with the secretion of proteins of the yeast Yarrowia lipolytica was site-directed mutagenesis,restricted and spliced to obtain a series of deletion mutants.These mutants would be useful for studing the function of different domains of TSR1 gene in yeast Yarrowia lipolytica

A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

Antimicrobial peptides are a class of small peptides with anti-extrogenous pathogen activities.They are derived from organism and possess antibacterial,antifungus,antiviruses and anticancer cell actions.In recent years,it’s found that some microbial pathogens are able to resist antimicrobial peptides.The constitutive and inducible mechanism of a pathogen resists a given peptide were reviewed in this paper.

Sixteen compounds have been isolated from the EtOAc fraction of 95% ethanolic extract of Sophora dunnii through silica gel, Sephadex LH-20 and semi-prerarative HPLC column chromatographies. Their structures were identified on the basis of NMR and MS spectra data as phaseollidin (1), L-maackiain (2), 2-(2',4'-dihidroxyphenyl)-5,6-methylenedioxy benzofuran (3), 8-demethyl-farrerol (4), liquiritigenin (5), genistein (6), 6-methylgenistein (7), 5-O-methyl genistein (8), 7,2',4'-trihydroxys-5-methoxy-isoflavanone (9), 7, 3', 4'-trihydroxy-isoflavanone (10), erythribyssin D (11), calycosin (12), trans-resveratrol (13), cis-resveratrol (14), stigmasterol (15), β-sitosterol (16). Among these, compounds 1-14 and 16 were isolated from this plant for the first time.
Chemical Fractionation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Sophora ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To evaluate the effect of gabapentin for treatment of neuropathic pain induced by bilateral sciatic nerve chronic constriction injury (bCCI) of rats.Methods Forty-eight female SPF Sprague-Dawley rats,weighing 180-200 g,aged 7-9 weeks,were randomly divided into 4 groups (n =12 each) using a random number table:control group (C group),sham operation group (S group),bCCI group,and bCCI + gabapentin group (G group).Gabapentin 100 mg/kg was intraperitoneally injected once at 15 min before operation and twice a day during 7-13 days after operation for 7 consecutive days.The bilateral mechanical paw withdrawal threshold (MWT),thermal paw withdrawal latency (TWL) and number of cold-stimulated paw withdrawal were measured before operation (baseline) and at 1,3,6,8,10,14 days after operation.Results Compared with C group,the bilateral MWT was significantly decreased,and the bilateral TWL was shortened in bCCI and G groups,and no significant change was found in MWT and TWL in S group.Compared with bCCI group,the bilateral MWT was significantly increased,the bilateral TWL was prolonged,and no significant change was found in the number of cold-stimulated paw withdrawal in G group.Conclusion Gabapentin can relieve thermal and mechanical hyperalgesia induced by bCCI,however,it exerts no effect on cold allodynia in rats.

This study was aimed to investigate the effect ofDanlou pills on prevent atherosclerosis from hypercholesterolemia rabbit and its relationship with inflammatory factors as well as PI3K/AKT signal pathways. A total of 24 Japanese male white rabbits were randomly divided into the control group (CL), model group (M) and Danlou group (DL), with 8 in each group. Normal diet was given to CL rabbits. High-fat diet was given to rabbits in other groups to establish the atherosclerosis model. Danlou pills (0.5 g·kg-1·d-1) were also given to DL rabbits. Rabbits were sacrificed after 9-week medication. The contents of blood lipid, TNF-α and IL-6 were detected. HE staining was used in the observation of histological changes in the aorta. Western blot was used to observe PI3K and p-AKT expression in the aorta. The results showed that compared with CL, the contents of TG, TC, LDL, IL-6 and TNF-α were significantly increased in M (P < 0.01); PI3K and p-AKT expression in the aorta were significantly decreased (P < 0.01). Compared with M, blood lipid, IL-6 and TNF-α were significantly reduced in DL (P < 0.05, orP < 0.01); PI3K and p-AKT expression were significantly increased (P < 0.01). It was concluded thatDanlou pills had prevention effects on atherosclerosis through reducing blood lipid and inflammatory factors. The action mechanism maybe related to the activation of PI3K/AKT signal pathways.

BACKGROUND: The surface electromyography can evaluate the effect of exercise by recording the parameters of muscle activities, and vibration exercise is the best supplement to traditional weight training. More research focuses on the effect of simple vibration training on the surface electromyography of lower limbs, but the influence of vibration combined with weight-bearing training is poorly understood.OBJECTIVE: To study the effect of vibration combined with load on the surface electromyography of lower limbs at the micro level of muscle working principle.METHODS: Eight healthy college students were recruited, and subjected to four different stimulations: vibration (45 Hz) combined with load (45% one repetition maximum (IRM)); vibration (45 Hz) combined with load (60% 1RM); vibration (50 Hz) combined with load (45% 1RM); vibration (50 Hz) combined with load (60% 1RM), followed by semi-squat exercise with the heel lifting, 10 times/minute, for 3 courses with more than 2 hours in between.RESULTS AND CONCLUSION: Different vibrations combined with loads made significant difference on the root mean square amplitude of the surface electromyography (P < 0.01), and there was a significantly increased root mean square amplitude in the vibration (50 Hz) combined with load (45% 1RM), especially at the medial gastrocnemius. The four kinds of stimulations made significant different effects on the surface electromyography of rectus femoris, tibialis anterior, and medial gastrocnemius, except semitendinosus (P ≤ 0.05). Moreover, the effect showed significant difference among different stimulations except vibration (50 Hz) combined with load (45% 1RM) (P ≤ 0.05). Compared with the other three stimulations, vibration (50 Hz) combined with load (45% 1RM) exerted better effect on the muscular activation. To conclude, different vibrations combined with loads exert different effects on the motor unit of same neuromuscular activity, and a suitable stimulation may produce better effect. Besides, the same stimulus for the motor unit of different neuromuscular activities produces different effects, which may match to the muscle nature.

OBJECTIVEGRP78 is a sensitive marker of endoplasmic reticulum stress. Caspase-12 is involved in apoptosis induced by endoplasmic reticulum stress. This study was designed to explore the changes of GRP78 and caspase-12 mRNA in neonatal rats with experimental hypoxic-ischemic white matter damage (WMD) and investigate the roles of endoplasmic reticulum stress in the WMD.

METHODSTwo-day-old rats were randomized to WMD and control groups (n=49 each). The pups were sacrificed at 0, 2, 4, 6, 12, 24 and 72 hrs after hypoxia-ischemia (HI). The light microscope was used to observe the brain pathological changes. Real time PCR was used to detect the expression of GRP78 mRNA and caspase-12 mRNA in the white matter tissue.

RESULTSThe expression of GRP78 mRNA began increasing 2 hrs after HI and peaked at 6 hrs in the WMD group, demonstrating significant differences at 2, 4, 6, 12, 24 and 72 hrs compared with the control group (P<0.05). The caspase-12 mRNA expression in the WMD group began increasing 6 hrs after HI and demonstrated significantly increased levels 6, 12 and 24 hrs after HI compared with those in the control group (P<0.05).

CONCLUSIONSGRP78 and caspase-12 mRNA expression increased significantly in neonatal rats with WMD. This suggests that endoplasmic reticulum stress may be induced following HI. Endoplasmic reticulum stress seems to be involved in the apoptosis of oligodendrocytes induced by HI in neonatal rats with WMD.

Animals ; Animals, Newborn ; Brain ; pathology ; Caspase 12 ; genetics ; Disease Models, Animal ; Heat-Shock Proteins ; genetics ; Hypoxia-Ischemia, Brain ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

BACKGROUND AND OBJECTIVEAs chemotherapy is generally used in the clinical treatment of cancer, the problem of multidrug resistance (MDR) of tumors against the chemotherapeutic agents becomes more and more serious. It has been the major cause for the failure of the chemotherapy. We detected the expressions of beta-catenin and tumor drug resistance related proteins, MRP2, P-gp, and Bcl-2, in esophageal squamous cell carcinoma (ESCC) to explore their function and correlation in the occurrence and development of MDR in ESCC.

METHODSWe used the tissue microarray technique, immunohistochemistry, and image analysis methods to detect the expressions of MRP2, P-gp, beta-catenin, and Bcl-2 proteins and analyze their relationships with clinical data in a ESCC tissue microarray consisting of 582 specimens of ESCC, 294 specimens of normal mucosa, 92 specimens of basal cell hyperplasia, and 87 specimens of dysplasia adjacent to cancer tissue.

RESULTSThe integral optical density (IOD) of MRP2 and Bcl-2, which was 195.7 +/- 175.9 x 10(3)) and 90.5 +/- 112.5 x 10(3)), respectively, was significantly higher in ESCC than in normal mucosa, which was 104.8 +/- 86.1 x103) and 25.2 +/- 46.6 x 10(3)), respectively (P < 0.01). The IOD of P-gp and beta-catenin, which was 57.7 +/- 75.5 x 10(3)) and 32.0 +/- 47.0 x 10(3)) respectively, was significantly lower in ESCC than in normal mucosa, which was 114.8 +/- 106.6 x 10(3)) and 46.1 +/- 35.7 x 10(3)), respectively (P < 0.01). According to the following order, well differentiated moderately differentiated poorly differentiated, the IOD of MRP2 increased in ESCC (P < 0.01). The IOD of beta-catenin was higher in poorly differentiated ESCC than in well or moderately differentiated ESCC (P < 0.01). The IOD of Bcl-2 was lower in well differentiated ESCC than in poorly and moderately differentiated ESCC (P < 0.01). The IOD of beta-catenin and Bcl-2 was higher in the ESCC of specimens with infiltration depths that were in membrane mucosa than those in the muscular layer or serous coat (P < 0.01). The IOD of Bcl-2 was significantly higher in cases with lymph node metastasis than in cases without (P < 0.01). Positive correlations which were respectively between the expressions of P-gp and MRP2, the expressions of P-gp and Bcl-2 were found (r = 0.288 and r = 0.253, respectively, P < 0.01).

CONCLUSIONSMRP2, P-gp, and Bcl-2 may be taken as prognostic factors for MDR of ESCC. beta-catenin may play an important role in carcinogenesis and progression of ESCC.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Multidrug Resistance-Associated Proteins ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Young Adult ; beta Catenin ; metabolism

OBJECTIVETo investigate the effect and mechanism (a selective cyclooxygenase-2 inhibitor) on invasive ability of human nasopharyngeal carcinoma (NPC) line CNE-2Z.

METHODSThe proliferation of NPC cells was examined by MTT assay. The invasive and migrating ability of NPC cells was detected with transwell chamber. E-cadherin protein expression was detected by immunocytochemistry and the expressions of Cox-2 and E-cadherin mRNA were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSMTT showed that celecoxib inhibited CNE-2Z proliferation in dose-dependent manner, the survival rate of cells treated with 25, 50, 100 µmol/L celecoxib (x(-) ± s) for 24 h was (94.75 ± 1.34)%, (91.77 ± 2.70)%, (64.54 ± 1.20)%, respectively, and the survival rate of cells treated for 48 h was (88.41 ± 1.28)%, (78.84 ± 1.56)%, (52.46 ± 2.25)%, respectively, the concentration of 50% inhibition concentration of a substance (IC50) was 100 µmol/L, the difference was statistically significant between different concentration groups in the same time-point (respectively, F were 462.204 and 1328.306, P < 0.01). Treated with different concentrations of celecoxib (0, 25, 50 µmol/L) for 24, the cell numbers (x(-) ± s) through PVPF by tumor invasion assay were (263.7 ± 13.5), (185.3 ± 8.7) and (144.0 ± 8.2), the difference was statistically significant between the experimental and control group (F = 102.089, P < 0.01). Immunocytochemistry showed that celecoxib significantly induced the increase of E-cadherin protein expression, also with a dose-dependence in 0 µmol/L, 25 µmol/L, 50 µmol/L group was (21.7 ± 2.6), (28.7 ± 2.4), (40.3 ± 1.3), and 50 µmol/L group increased significantly (F = 78.637, P < 0.01). RT-PCR showed that celecoxib reduced the expression of Cox-2 mRNA expression in 25, 50 µmol/L group decreased significantly compared with the control group (respectively, t were 23.950 and 36.651, P < 0.01), but it enhanced the expression of E-cadherin mRNA expression in 25, 50 µmol/L group was significantly higher (respectively, t were 35.829 and 81.497, P < 0.01).

CONCLUSIONCelecoxib can inhibits the invasive ability of NPC cell line CNE-2Z, which possibly relates with the upregulated expression of E-cadherin.

Apoptosis ; drug effects ; Cadherins ; genetics ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Celecoxib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Pyrazoles ; pharmacology ; Sulfonamides ; pharmacology