Altitude ; Animals ; Blood Glucose ; metabolism ; Cholesterol ; blood ; Hypoxia ; metabolism ; Mice ; Weight Loss

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

To enhance the quality and efficiency of ozagrel by investigating the differences between the ozagrel polymorphs in bioavailability. Solid ozagrel in different polymorph forms were orally administered to SD rats. An HPLC method was established to determinate plasma level of ozagrel. The bioavailabilities of two polymorph forms were calculated and compared. The pharmacokinetic parameters of ozagrel, were as follows: Cmax was 32.72 ± 17.04 and 34.01 ± 19.13 mg · L(-1), respectively; AUC0-t was 61.14 ± 14.76 and 85.56 ± 18.08 mg · L(-1) · h, respectively; t½ was 1.53 ± 0.51 and 4.73 ± 3.00 h, respectively. There was no significant difference in pharmacokinetic parameters between form I and II polymorphs of ozagrel while the t½ of form II is longer, which indicates that the use of form II polymorph as pharmaceutical product may prolong the effective action time in clinics. This would help the polymorph quality control in drug production.
Animals ; Biological Availability ; Chromatography, High Pressure Liquid ; Methacrylates ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

The viral RNA was extracted from purified cymbidium mosa ic virus (CyMV) isolated from Dendrobium orchid cultivated in Hainan island. The gene of the movement protein (MP) was amplified by means of reverse transcripti on-polymerase chain reaction (RT-PCR), and cloned into pGEM-T easy vector. Se quence analysis showed that the gene fragment contained 3 open reading frames (O RFs) which may be encoding 14 kD、12 kD and 10 kD peptides. The nucleotide seque nce of the cloned gene fragment shared 97.8% homology with the MP genes of CyMV isolated from orchids cultivated in Hawaii and Singapore.

The present investigation was undertaken in order to select the surface-sterilization technique most efficient for eliminating epiphytes, to document endophytes of healthy tissues from Glaycyrrhiza inflat Bat. in Xinjiang. Surface sterilization with 5% commercial solution of sodium hypochlorite for 5 minutes was reaffirmed as adequate for removing epiphytes on licorice roots. From the 151 segments incubated, 149 bacterial isolates and 2 fungal isolates were obtained. From all the isolates, Bacterial isolates were identified by VITEK-AMS. Part of Bacteria were identified in 13 different genus. Fungal species were characterized as Penicillium sp. and Fusarium sp.with microscope.

Objective: To observe the clinical effect of Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method versus ordinary acupuncture on superficial fascia for treating neck type cervical spondylopathy in Kyrgyz. Methods:A total of 64 Kyrgyz patients conforming to the diagnostic criteria of neck type cervical spondylopathy were included. The patients were randomized into a Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture group and a conventional acupuncture group, with 32 cases in each group. Patients in the Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture group were treated with Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method to stimulate the superficial fascia, the stimulation sites were primarily located at bilateral sides of the cervical vertebra as well as the trigger points in shoulder-neck region; patients in the conventional acupuncture group were punctured at the same acupoints, with twirling reducing method, and the depth of insertion was determined by the treated region. Patients in both groups received treatment every day for a succession of 5 d as a course, with a 2-day interval between 2 courses, and the whole treatment lasted for 3 courses. After 3 courses of treatment, the McGill pain questionnaire (MPQ) and neck disability index (NDI) were measured to compare the clinical effect between the two groups. Results: After treatment, scores of MPQ and NDI scale dropped when compared with those before treatment, and the differences showed statistical significance (allP<0.05); scores of MPQ and NDI in the Jin Gou Diao Yu (gold-hook-fishing) acupuncture group were substantially lower than those in the conventional acupuncture group, and the differences showed statistical significance (allP<0.05). The total effective rate was 96.9％ in the Jin Gou Diao Yu (gold-hook-fishing) acupuncture group and the cured rate was 78.1％, which were higher than 81.3％ and 40.6％ in the conventional acupuncture group, and the differences showed statistical significance (bothP<0.05). Conclusion: Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method by stimulating superficial fascia to treat cervical spondylopathy in Kyrgyz can produce a better clinical effect than conventional acupuncture treatment, and is effective in improving pain and stiffness in patients, and thus is worth clinical popularization.

Abstract: Objective　To investigate the distribution and drug resistance characteristics of pathogenic bacteria in patients with neutropenic acute leukemia (AL) and bloodstream infections (BSI). Methods　The clinical data of 258 neutropenic acute leukemia patients with bloodstream infections, who admitted to Shengjing Hospital of China Medical University from January 2016 to December 2021, were collected and analyzed for pathogenic bacteria and drug resistance. Results A total of 268 strains of pathogenic bacteria were isolated from 258 patients, including 180 strains of gram-negative bacteria (67.16%), 61 strains of gram-positive bacteria (22.76%), and 27 strains of fungi (10.07%). Gram-negative bacteria were mainly Klebsiella pneumoniae (53/268, 19.78%), Escherichia coli (49/268, 18.28%) and Pseudomonas aeruginosa (41/268, 15.30%). Gram-positive bacteria were mainly coagulase negative Staphylococcus (31/268, 11.57%) and Staphylococcus aureus(17/268, 6.34%). The main fungi were Candida tropicalis (25/268, 9.33%). Escherichia coli (33/268, 12.31%) was the most common pathogen isolated from acute myeloid leukemia (AML), followed by Pseudomonas aeruginosa (25/268, 9.33%), coagulase-negative Staphylococcus (18/268, 6.72%) and Candida tropicalis (18/268, 6.72%). Klebsiella pneumoniae (35/268, 13.06%) was the most common pathogen isolated from acute lymphoblastic leukemia (ALL),followed by Pseudomonas aeruginosa (15/268, 5.60%) and Escherichia coli (14/268, 5.22%). The resistance of Gram-negative bacteria to piperacillin/tazobactam, cefoperazone/sulbactam, imipenem, meropenem, ertapenem, amikacin, cefoxitin, amoxicillin/clavulanic acid was low. Gram-positive bacteria were sensitive to linezolid and vancomycin. Candida was sensitive to flucytosine, amphotericin B and itraconazole. Conclusions　In patients with granulosa after AL chemotherapy combined with BSI, the pathogenic bacteria isolated from AML are diverse, and the pathogenic bacteria isolated from ALL are mainly gram-negative bacteria. Pathogenic bacteria have different degrees of drug resistance to commonly used antibacterial drugs, so it is important to strengthen the monitoring of the distribution of pathogenic bacteria and the change of drug resistance and rational use of antibacterial drugs to minimize the death of patients.

Experiments were made on 6 adult cats. Brainstem neurons that project to the phrenic nucleus were retrogradely labeled by microinjection of WGA-HRP into phrenic nucleus. The brainstem sections were doubly processed for HRP histochemical staining with the TMB-Co-DAB method, and GABA immunohistochemical staining with anti-GABA primary antibody and FITC conjugated immunofluorescent secondary antibody. HRP-FITC double-labeled neurons were observed in the pontine K(o)lliker-Fuse nucleus and areas around the retrofacial nucleus (B(o)tzinger complex). Double-labeled neurons were also observed in the raphe nucleus, the lateral paragigantocellular nucleus, and the vestibular nucleus. The results show that brainstem GABAergic neurons in these structures, especially those in the K(o)lliker-Fuse nucleus and the B(o)t.C, send axonal projections to the phrenic nucleus.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the beta-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.
Animals ; Cells, Cultured ; Chickens ; Cloning, Molecular ; methods ; Fallopian Tubes ; metabolism ; Female ; Organ Specificity ; Ovalbumin ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; Transfection ; methods ; Women

Based on the 2 x 2 contingency table, by using multi-species relevance (variance ratio, VR), chi2-test, Ochiai index, Dice index, Jaccard index, t-test of v/x and F-test of Morisita, s index, the interspecific relationships and the spatial distribution pattern between 20 dominants in Kangding Zheduo Mountain of Sichuan province were studied. The results indicated that the interspecific association between dominants of Sinopodophyllum hexandrum community in this area did not show significant association, which suggested that the S. hexandrum community was in mature stage, and showed stronger independency, among total 190 pairs in 20 dominant species, 2 species pairs exhibited extremely significantly positive association, 12 species pairs showed significantly positive association, 6 species pairs exhibited significantly negative association and there were no pairs showed extremely significantly negative association. S. hexandrum in community did not show significant association, which indicates they are independent in community, the spatial distribution pattern of S. hexandrum is characterized by random distribution.
Berberidaceae ; classification ; growth & development ; Biodiversity ; China ; Ecosystem ; Endangered Species ; Plants, Medicinal ; classification ; growth & development