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

BACKGROUNDLeukocyte adhesion deficiency type 1 (LAD-1) is a rare, autosomal recessive inherited immunodeficiency disease characterized by recurrent severe bacterial infection, impaired pus formation, poor wound healing, associated with the mutation in the CD18 gene responsible for the ability of the leucocytes to migrate from the blood stream towards the site of inflammation. Correct and early diagnosis of LAD-1 is vital to the success of treatment and prevention of aggressive infections. The purpose of this study was to collect the clinical findings of the disease and to identify the genetic entity.

METHODSCD18 expression in the peripheral blood leukocytes from the patient, his parents and normal control was measured with flow cytometry. The entire coding regions of the CD18 gene were screened with direct sequencing genomic DNA.

RESULTSCD18 expression level on this patient's leukocyte surface was significantly decreased, with normal level in control group, his father and mother. Gene analysis revealed that this patient had a homozygous c.899A > T missense mutation in exon 8 of CD18 gene, causing the substitution of Asp to Val at the 300 amino acid. His parents were both heterozygous carriers while no such mutation was found in 50 normal controls.

CONCLUSIONThis study disclosed a novel point mutation Asp 300 Val located in a highly conserved region (HCR) of CD18 and confirmed the heterogeneity of the mutations causing LAD-1, indicating it was quite beneficial to establish correct and early diagnosis in children with severe LAD-1.

Asian Continental Ancestry Group ; CD18 Antigens ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Flow Cytometry ; Humans ; Leukocyte-Adhesion Deficiency Syndrome ; etiology ; genetics ; Male ; Point Mutation ; genetics ; Polymerase Chain Reaction

OBJECTIVETo investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).

METHODSWAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.

RESULTSWAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).

CONCLUSIONSThe WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.

Adolescent ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Proliferation ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Female ; Humans ; Infant ; Jurkat Cells ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; etiology ; metabolism ; RNA, Messenger ; analysis ; Wiskott-Aldrich Syndrome Protein Family ; analysis ; genetics ; physiology