By exploring the different components of the lysis buffer and optimize the 2-DE conditions,established the best proteomics technical system for Populus euphratica's heteromorphic leaves,while take the heteromorphic leaves in the same blanche as the test materials to find differences between the protein expressions of the leaves.It showed that the lysis solution which containing 2mmol/L thiourea,7mmol/L urea,2% CHAPS,60mmol/L DTT and 0.2% IPG buffer could dissolve the protein better.Through tandem mass spectrum,the results show that heteromorphic leaves are different in photosynthesis and respiration.This research offered valuable informations for understanding the molecular mechanism during leaves development and elucidating the mechanism of the eco-adaptability of Populus euphratica.

Objective To investigate the expression of steroidogenic factor 1 ( SF-1 ) in eutopic endometrium,ovarian endometriosis (EM) and adenomyosis.Methods From Jan.2008 to Dec.2010,30 patients with both ovarian endometriotic cysts and adenomyosis underwent hysterectomy,cystectomy or adenxectomy in Peking University First Hospital.The combination of ovarian EM and denomyosis were confirmed in 17 cases.In the mean time,endometria form 10 patients with cervical intraepithelial neoplasm ( CIN ) Ⅲ underwent hysterectomy were selected as controls.The expression of SF-l in eutopic endometrium,ovarian EM and adenomyosis was detected by immunohistochemistry staining.Results No immunoreactivity of SF-l was observed glandular cell and stromal cell in eutopic endometrium of both groups.14/17 of positive rate of SF-l were observed in lesions of ovarian endometriosis.SF-l protein was not expressed in the endometrial glandular cells of ovarian EM and in adenomyosis foci.Conclusion The differences of SF-l expression in ovarian endometrioma and adenomyosis foci might mediate the development of the disease.

Animals ; Animals, Newborn ; Cerebral Cortex ; drug effects ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P＜0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P ＜ 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.

There is no specific treatment for post-polio syndrome. The common applied therapies include mediciation, exercise, cogni-tive behavioural therapy, physiotherapy, occupational therapy, Traditional Chinese Medicine, assistive technology, psychological and social factors adjustment, interdisciplinary comprehensive rehabilitation, and so on.

Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
3T3 Cells ; Animals ; Biocompatible Materials ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Durapatite ; chemistry ; Lysostaphin ; pharmacology ; Materials Testing ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Microscopy, Electron, Scanning ; X-Ray Diffraction

Yeast strains with improved ethanol yield are important for efficient bioconversion of lignocellulosic biomass for fuel ethanol.Candida shehatae CICC1766 was adapted to 4%(v/v)ethanol,and then subjected to UV mutagenesis.One respiration deficient mutant Rd-5 with improved xylose fermentation capability was selected.Protoplasts of Rd-5 were inactivated by UV treatment,followed by the PEG-mediated protoplast fusion with a Saccharomyces cerevisiae strain with good ethanol-fermenting capability.The xylose fermenting capability of the fusants was investigated,and the fusant F6 demonstrated good ethanol fermentation performance,producing 18.75g/L ethanol from 50g/L xylose with an ethanol yield of 0.375 or 73.4% of its theoretical value of 0.511.Comparing with its parent Candida shehatae strain,the ethanol yield of F6 was increased by 28%.

Cerebral ischemia is due to cerebral blood supply disorders caused by ischemia and hypoxia resulting in localized ischemic brain necrosis or brain softening of the disease, leading to irreversible brain damage and subsequent loss of neuronal function is a serious threat to human health One of several diseases. For patients with cerebral ischemia, often the lack of effective and extensive treatment. In addition, cerebral ischemia with morbidity, morbidity and mortality are characterized by high, so by the medical profession at home and abroad attention. As a traditional Chinese medicine, cordyceps sinensis (CS) is a complex of ergot fungus, which is parasitized on the larvae of the bat-moth family. The compound is composed of cordycepin, cordyceps polysaccharide, cordyceps sinensis peptides, ergosterol, mannitol, fatty acids and trace elements such as a variety of ingredients, with a wide range of pharmacological effects. Over the years, domestic and foreign scholars on the pharmacological effects of cordyceps sinensis were more comprehensive study of its prevention and treatment of cerebral ischemia is also deepening, found that cordyceps sinensis on cerebral ischemia with anti-inflammatory, reduce oxidative damage and neuronal ischemia damage, reduce neuronal apoptosis, improve memory cognition, reduce thrombosis, inhibit NO production, enhance mitochondrial energy supply, scavenging free radicals and other prevention and treatment. But no relevant review. In this paper, the domestic and foreign literatures on the prevention and treatment of cerebral ischemia by cordyceps sinensis were summarized, analyzed and summarized in order to provide useful information for the research and further development of cordyceps sinensis.

OBJECTIVETo observe the dynamic expression of calcium-sensing receptor(CaSR) in myocardium of diabetic rats.

METHODSThirty male Wistar rats were randomly divided into 3 groups including control, diabetic-4 week and diabetic-8 week groups(n = 10). The type 2 diabetes mellitus models were established by intraperitoneal injection of streptozotocin (STZ, 30 mg/kg) after high-fat and high-sugar diet for one month. The cardiac morphology was observed by electron microscope. Western blot analyzed the expression of CaSR, phospholamban (PLN), a calcium handling regulator, and Ca+-ATPase(SERCA) in cardiac tissues.

RESULTSCompared with control group, the expressions of CaSR and SERCA were decreased, while the expression of PLN was significantly increased in a time-dependent manner in diabetic groups. Meanwhile diabetic rats displayed abnormal cardiac structure.

CONCLUSIONThese results indicate that the CaSR expression of myocardium is reduced in the progression of DCM, and its potential mechanism may be related to the imnaired intracellular calcium homeostasis.

Animals ; Calcium-Binding Proteins ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; Diabetic Cardiomyopathies ; metabolism ; physiopathology ; Disease Progression ; Heart ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Receptors, Calcium-Sensing ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Streptozocin

OBJECTIVETo clone the p40 and p35 subunit cDNA of porcine IL-12(pIL-12) and construct the fusion gene of recombinant porcine single-chain interleukin-12 (pscIL-12).

METHODSThe total RNAs were extracted from porcine peripheral blood mononuclear cells (PBMCs) and porcine splenic lymphocytes for cloning pIL-12 p35 and p40 cDNA by RT-PCR. A hydrophobic polypeptide linker (Gly4Ser)3 was used for splicing two different gene fragments (pIL-12) p40+linker+p35 (pscIL-12) by recombinant PCR to construct pscIL-12 fusion gene. The pscIL-12 fusion gene was then inserted into pcDNA3.1(+) eukaryotic expression plasmid, and the resulted pcDNA3.1(+)-pscIL-12 was transfected into CHO-K1 cells via lipofectin. The expression of pscIL-12 mRNA in the transfected cells was identified by RT-PCR.

RESULTSThe sequence of the cloned porcine IL-12 p40 and p35 cDNA and the constructed pscIL-12 fusion gene were verified by PCR, restriction enzyme digestion and sequencing. The mRNA of pscIL-12 fusion gene was detected in the transfected CHO-K1 cells by RT-PCR.

CONCLUSIONThe constructed pcDNA3.1(+)-pscIL-12 eukaryotic expression plasmid allows expression of pscIL-12 in CHO-K1 cells, thus facilitating further study of the biological activity and adjuvant effect of pscIL-12 fusion protein.

Animals ; Base Sequence ; CHO Cells ; Cloning, Molecular ; Cricetinae ; DNA, Complementary ; genetics ; Interleukin-12 ; biosynthesis ; classification ; genetics ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Swine