Gastrointestinal Tract ; physiopathology ; Humans ; Infection ; physiopathology ; Sepsis ; physiopathology ; Shock, Septic ; physiopathology

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

Abnormalities, Multiple ; diagnosis ; genetics ; pathology ; Brain ; diagnostic imaging ; pathology ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Optic Atrophies, Hereditary ; diagnostic imaging ; pathology ; Radiography ; Septo-Optic Dysplasia ; diagnosis ; genetics ; pathology ; Septum Pellucidum ; diagnostic imaging ; pathology

OBJECTIVETo study the influence of humic acid fertilizer on plant growth, assimilation base, dried biomass accumulation, yield, quality and disease infection of Angelica sinensis.

METHODThree kinds of humic acid fertilizer and an amino acid liquid fertilizer were tested in randomized groups at 1 level with 3 times repeat.

RESULTT1 promoted plant and root growth effectively, increased dried biomass accumulation and fresh root average weight remarkably, the yield was increased, the content of ethanol extract was increased by 11.31%. T3 promoted plant and root growth quickly, enlarged leaves area and increased dried plant weight, but effect lasted shortly, the content of ethanol extract was increased by 5.23%. T4 increased more leaves in late growth period, enlarged leaves area, the yield was increased, the content of ethanol extract was increased by 3.09%. T2 increased fresh root average weight remarkably, the yield was increased.

CONCLUSIONHumic acid fertilizer and amino acid liquid fertilizer could effectively promote plant growth, enlarge leaves area, promote dried biomass accumulation and transformation to root and increase yield and content of ethanol extract effectively.

Angelica sinensis ; growth & development ; metabolism ; Biomass ; Fertilizers ; Humic Substances ; Plant Leaves ; growth & development ; metabolism ; Plant Roots ; growth & development ; metabolism

The steps for installation and withdrawal of WYD2000 field surgical lamp were introduced.The failures and causes of broken cross-arm connector of WYD2000 field surgical lamp were analyzed.The problems of WYD2000 field surgical lamp in vulnerability to breaking and difficulty in maintenance were solved by designing and manufacturing a special maintenance tool and optimizing the materials and fixing mode of cross-arm connection.References were provided for main-tenance and improvement of WYD2000 field surgical lamp.[Chinese Medical Equipment Journal,2024,45(4):116-118]

Adult ; Diagnosis, Differential ; Female ; Humans ; Ovarian Neoplasms ; pathology ; Sertoli-Leydig Cell Tumor ; pathology

OBJECTIVETo study the effect of methylene blue combined with aprotinin on intraperitoneal adhesion.

METHODSFrom May 2000 to February 2004, there were 83 patients receiving total or partial colectomy and temporary ileostomy or colostomy, and second anastomosis was performed within 8-12 weeks after the first operation. These patients were divided into four groups and followed by intraperitoneal administration of saline,methylene blue,aprotinin,combined methylene blue and aprotinin respectively during the second operation, then adhesion formation was quantitatively graded.

RESULTSThe adhesion rate was 15% in combination group, 83% in saline group, 40% in methylene blue group, and 45% in aprotinin group, respectively. The adhesion rate was significantly lower in combination group(P< 0.05).

CONCLUSIONSMethylene blue and aprotinin can decrease the incidence of intraperitoneal adhesion significantly. The combination of these two drugs has significant effectiveness in the treatment of intraperitoneal adhesion.

Aprotinin ; therapeutic use ; Drug Therapy, Combination ; Female ; Humans ; Methylene Blue ; therapeutic use ; Peritoneal Diseases ; pathology ; prevention & control ; Postoperative Complications ; prevention & control ; Tissue Adhesions ; prevention & control

OBJECTIVETo acquire lots of cell to culture during the primary cell culture.

METHODWe take the split-thickness skin slide technique to acquire the dissociated fibroblast cell in two big-ear rats.

RESULTSThe cell number is above 10(6) from 1 cm x 2 cm split-thickness skin slide and the technique is simple, economic, effectve.

CONCLUSIONWe think this way is better than other methods, and should be adopted in the primary cell culture, especially in fibroblast transplantation by injection.

Animals ; Cell Culture Techniques ; methods ; Fibroblasts ; cytology ; transplantation ; Rabbits

AIM:To explore the role of aloperine in ischemia-reperfusion(I/R)-induced H9c2 cardiomyocyte injury and inflammation.METHODS: The H9c2 cardiomyocytes were cultured under hypoxia and re-oxygenation condi-tions to simulate ischemia-reperfusion(SI/R)injury.After treatment with aloperine at various doses,the cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.Simultaneously,the levels of lactate dehydro-genase(LDH),malonaldehyde(MDA)and caspase-3 activity were detected by the commercial kits.The levels of inflam-matory cytokines were also detected by ELISA.Moreover,the effects of aloperine on the activation of PI 3K/AKT signaling pathway were determined by Western blot.RESULTS:Pre-treatment with aloperine remarkably abated the inhibitory effect of SI/R on H9c2 cell viability,and decreased the elevations of LDH and MDA triggered by SI /R(P<0.05).Pre-treat-ment with aloperine dramatically suppressed the cell apoptosis induced by SI /R treatment(P<0.05), concomitant with the decrease in caspase-3 activity and increase in Bcl-2/Bax ratio(P<0.05).In contrast to SI/R group,aloperine treat-ment notably restrained the concentrations of pro-inflammatory cytokines, including interleukin-6, tumor necrosis factor-α and interleukin-1β(P<0.05).Furthermore, aloperine remarkably increased the protein levels of p-PI3K and p-AKT. While blocking the PI3K/AKT pathway with its specific inhibitor LY294002, the viability-promoting, anti-apoptotic and anti-inflammatory effects of aloperine on the H 9c2 cells were obviously attenuated(P<0.05).CONCLUSION: Alope-rine protects against cardiomyocytes from I/R injury and inhibits inflammatory responses by activating the PI 3K/AKT signa-ling pathway,implying a potential benefic role of aloperine against myocardial I /R injury.

MicroRNAs have been identified as a new class of regulatory molecules that affect many biological functions by interferring the target gene expressions. Latest studies demonstrate that microRNAs can influence many pivotal bio-processes and deeply involve in the metabolism of glucose, lipid and amino acid and biological oxidation. For glucose metabolism, microRNAs are related to insulin secretion, insulin sensitivity, glucose uptake, glycolysis, oxidation and mitochondrial function. For lipid matebolism, microRNAs can regulate the target genes related to lipid biosynthesis, catabolism and transportation. MicroRNAs can influence glutamine catabolism.
Animals ; Glucose ; metabolism ; Glutamine ; metabolism ; Humans ; Insulin ; metabolism ; Insulin Secretion ; Lipid Metabolism ; physiology ; Metabolism ; physiology ; MicroRNAs ; physiology