BACKGROUNDMutations in mitotic checkpoint genes have been detected in several human cancers, which exhibit chromosome instability. We wanted to know whether mutation of hBub1 could occur in transformed human embryo lung fibroblasts (HELF) cells induced by a chemical carcinogen.

METHODSHELF cells were transformed by N-methyl-N'-nitro-N-nitrosoguaridine (MNNG), and three flasks of transformed HELF cells (named as T1, T2, and T3) were selected as amplifiers, and mutations of hBub1 in these transformed cells were analyzed by PCR-SSCP and sequencing.

RESULTSIt was found that any one of three transformed cell lines exhibited aneuploidy with a low mitotic checkpoint function. Subsequent PCR-SSCP and sequence analysis showed an AGT to CGT or ATT mutation at codon 80 in hBub1 gene in T1 cells with a resultant change in amino acid sequence.

CONCLUSIONOur study demonstrated that the mitotic checkpoint genes could be targets of MNNG.

Cell Line, Transformed ; Chromosome Aberrations ; Down-Regulation ; Fibroblasts ; drug effects ; Humans ; Lung ; cytology ; Methylnitronitrosoguanidine ; toxicity ; Mitosis ; drug effects ; Mutation ; Protein Kinases ; genetics ; Protein-Serine-Threonine Kinases

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.

OBJECTIVETo observe and compare perioperative myocardial enzyme changes in 107 patients with congenital (CHD, n = 53), rheumatic (RHD, n = 40) and coronary artery (CAD, n = 14) diseases, and to find whether different diseases can affect the release and recovery of myocardial enzymes after heart operations.

METHODSOn the day before operation and the 1st, 3rd, 5th and the 8th day after operation, the venous blood was taken to measure the release of myocardial enzymes: aspartate aminotransferase (AST), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH) and LDH-1.

RESULTSAll the enzymes measured before operation in three groups were in the normal range; their release increased abruptly on the 1st day postoperatively to 2 - 15 times of those before operation; on the 3rd day, they recovered to some degrees, and on the 8th day they recovered to normal in all groups except LDH and LDH-1 in rh and CAD groups. Because the aortic cross-clamp time (CCT) had a good positive correlation to the release of myocardial enzymes, those patients whose CCT was over 60 minutes in three groups were compared revealing that the CCT was not different between three groups (P < 0.05). The release of CK, CK-MB and AST was significantly higher in CHD60 group than those in CHD60 and CAD60 groups, they recovered afterwards; while the release of DH and LDH-1 was higher in CAD60 group than those in CAD60 and in CHD60 groups from the 1st day to the 8th day postoperatively.

CONCLUSIONSThe release of all the 5 enzymes measured before operation was in normal range in selected CHD, RHD and CAD patients. The release peak and the recovery order of all enzymes were the same in three groups. The release of CK, CK-MB and AST was higher in CHD60 group than those in RHD60 and CAD60 groups on the 1st day. The release of LDH and LDH-1 was higher in RHD60 group than those in CHD60 and CAD60 groups from the 1st day to the 8th day postoperatively. The shorter the CCT is, the less the release of myocardial enzymes. Using the release of LDH and LDH-1 to evaluate the recovery of myocardial injury after open-heart operations was recommended.

Adolescent ; Adult ; Aspartate Aminotransferases ; blood ; Child ; Coronary Artery Bypass ; Coronary Artery Disease ; blood ; enzymology ; surgery ; Creatine Kinase ; blood ; Creatine Kinase, MB Form ; blood ; Female ; Heart Defects, Congenital ; blood ; enzymology ; surgery ; Humans ; Intraoperative Period ; Isoenzymes ; blood ; L-Lactate Dehydrogenase ; blood ; Male ; Middle Aged ; Myocardium ; enzymology ; pathology ; Rheumatic Heart Disease ; blood ; enzymology ; surgery ; Time Factors

Objective: To observe the effect of L-carniti ne (L-CN) in the treatment of congestive heart failure (CHF). Meth ods: Fifty-six cases of chronic CHF randomly received routine treatment (Digitalis, diuretics, vasodilator, ACEI or βblocker) or L-CN (3.0 g/d ,V D×10 d) with routine therapy. Results: The treatment efficiency of L-CN group and control group were 89.3% and 60.7% (P<0.01), respect ively. No adverse reactions related to the drug were observed. Conclusio n: L-CN with routine therapy might be a safe way to the treat CHF.

OBJECTIVETo reassess the natural history of polycythemia vera (PV) in Chinese and evaluate the relationship between the incidence of thrombosis, post-polycythaemic myelofibrosis with myeloid metaplasia( PPMM) , leukemia transformation and the therapeutic outcome and prognostic factors.

METHODSThe clinical manifestations, laboratory parameters and treatment were retrospectively analyzed in 287 patients with PV. Univariate analysis of prognostic factors was performed using Log-rank model and multivariate analysis using COX model in term of the incidence of thrombosis, PPMM, hematologic or non hematologic cancers and mortality.

RESULTSOf the 287 patients, the median follow-up time was 46 (8-360) months. 208 thromboses were recorded in 115 patients. Twice or more thrombotic events occurred on 59 patients (51.34%). Most of these episodes occurred either at presentation or in the 2 years before diagnosis. Elder patients, prior thrombosis, poor response to therapy were associated with poor prognosis. With these three adverse prognostic factors, the patients could be separated into different risk groups. The incidence of thrombosis was higher in high risk group. 36 patients progressed to PPMM, the median time to PPMM was 80 (7-190) months. Higher WBC count, splenomegaly and treatment with alkylating agent and hydroxy-carbamide (HU) were associated with poor prognosis. 2 cases progressed to AML. 1 to lymphoma and 1 to nonhematologic cancer. 13 patients died, the cause of death was fatal thrombosis in 9 and AML in 2.

CONCLUSIONThe incidence of thromboembolism is higher and the time to myelofibrosis was shorter in Chinese PV patients than in western PV patients. The main factors that influence the survival of PV patients are thromboembolism and leukemia transformation.

Acute Disease ; Female ; Follow-Up Studies ; Humans ; Leukemia ; etiology ; Male ; Polycythemia Vera ; complications ; Primary Myelofibrosis ; etiology ; Prognosis ; Risk Factors ; Thromboembolism ; etiology

Chronic enteritis can produce an excess of reactive oxygen species resulting in cellular damage. Stanniocalcin-1(STC-1) reportedly possesses anti-oxidative activity, the aim of this study was to define more clearly the direct contribution of STC-1 to anti-oxidative stress in cattle. In this study, primary intestinal epithelial cells (IECs) were exposed to hydrogen peroxide (H2O2) for different time intervals to mimic chronic enteritis-induced cellular damage. Prior to treatment with 200 microM H2O2, the cells were transfected with a recombinant plasmid for 48 h to over-express STC-1. Acridine orange/ethidium bromide (AO/EB) double staining and trypan blue exclusion assays were then performed to measure cell viability and apoptosis of the cells, respectively. The expression of STC-1 and apoptosis-related proteins in the cells was monitored by real-time PCR and Western blotting. The results indicated that both STC-1 mRNA and protein expression levels positively correlated with the duration of H2O2 treatment. H2O2 damaged the bovine IECs in a time-dependent manner, and this effect was attenuated by STC-1 over-expression. Furthermore, over-expression of STC-1 up-regulated Bcl-2 protein expression and slightly down-regulated caspase-3 production in the damaged cells. Findings from this study suggested that STC-1 plays a protective role in intestinal cells through an antioxidant mechanism.
Animals ; Animals, Newborn ; Blotting, Western/veterinary ; Caspase 3/*genetics/metabolism ; Cattle ; Cattle Diseases/etiology/*genetics/metabolism ; Duodenum/metabolism ; Enteritis/etiology/genetics/metabolism/*veterinary ; Epithelial Cells/metabolism ; *Gene Expression Regulation ; Glycoproteins/*genetics/metabolism ; Hydrogen Peroxide/pharmacology ; Male ; Proto-Oncogene Proteins c-bcl-2/*genetics/metabolism ; RNA, Messenger/genetics/metabolism ; Real-Time Polymerase Chain Reaction/veterinary

This study aimed to report the clinical characteristics and COMP gene mutation of a family with pseudoachondroplasia (PSACH), a relatively rare spinal and epiphyseal dysplasia that is inherited as an autosomal dominant trait. Clinical information on a 5-year-2-month-old PSACH child and his parents was collected and analyzed. Diagnosis was confirmed by PCR amplification and direct sequencing of all the 19 exons and their flanking sequences of COMP gene, and the mutation was further ascertained by cloning analysis of exon 10. The child presented with short and stubby fingers, bow leg, short limb dwarfism and metaphysic broadening in long bone as well as lumbar lordosis. A mutation c.1048_1116del (p.Asn350_Asp372del) in exon 10, inherited from his father who did not demonstrate any phenotypic feature of PSACH, was detected in the child. PSACH was diagnosed definitively by means of COMP mutation analysis, on the basis of the child's clinical and imaging features. The non-penetrance phenomenon of COMP mutation was described for the first time in PSACH.
Achondroplasia ; genetics ; Cartilage Oligomeric Matrix Protein ; genetics ; Child, Preschool ; Cloning, Molecular ; Humans ; Male ; Mutation

OBJECTIVETo investigate the normal range of (CAG)n in spinocerebellar ataxia type 1 (SCA1) gene and spinocerebellar ataxia type 3 (SCA3/MJD) gene in 110 normal subjects of Han population in Northeastern China, to assess the genotypes for clinically diagnosed spinocerebellar ataxia(SCA) individuals including 25 patients from 8 families and 6 sporadic patients, and to make presymptomatic and prenatal diagnosis.

METHODSDNA fragments from the normal subjects and the patients were detected by fluorescence-PCR. Homozygosities were selected for DNA sequencing.

RESULTSThe normal ranges of (CAG)n of SCA1 and SCA3/MJD were 20-39 and 14-38 repeats respectively, SCA1 was found mostly to be 26 and 27 repeats, allele frequency 34.09% and 20.91%; heterozygosity was 84.55%, SCA3/MJD was found mostly to be 14 repeats, allele frequency 39.55%, heterozygosity was 78.18%.(CAG)(68) of SCA3/MJD gene of one affected individual had been found in a family but no CAG mutative expansion in related members was observed.

CONCLUSIONThe normal ranges of CAG repeats vary with areas and races. SCAs genotyping is the first choice in presymptomatic and prenatal diagnosis.

Ataxin-1 ; Ataxin-3 ; Ataxins ; China ; DNA ; chemistry ; genetics ; Family Health ; Female ; Gene Frequency ; Genotype ; Humans ; Machado-Joseph Disease ; diagnosis ; genetics ; Male ; Nerve Tissue Proteins ; genetics ; Nuclear Proteins ; genetics ; Pedigree ; Repressor Proteins ; Sequence Analysis, DNA ; Spinocerebellar Ataxias ; diagnosis ; genetics ; Trinucleotide Repeat Expansion ; genetics ; Trinucleotide Repeats ; genetics

OBJECTIVETo investigate the effect of penehyclidine hydrochloride (PHC) in a rat model of renal injury induced by hemorrhagic shock and lipopolysaccharides (LPS).

METHODSForty-five healthy Wistar rats were randomized into sham operated group, model group, and 3 penehyclidine hydrochloride (PHC) dose (1, 2 and 3 mg/kg) groups (PHC1, PHC2, and PHC3 groups, respectively). The arterial blood samples were collected to determine the concentrations of serum tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-1 (IL-1), urine creatinine (Cr) and blood urine nitrogen (BUN), and the renal tissues were collected to measure the expressions of ICAM-1 and nuclear factor-κB (NF-κB) and observe the pathological changes.

RESULTSTNF-α, IL-8, IL-1, Cr, BUN, ICAM-1 and NF-κB in the 3 PHC groups were significantly lower than those in the model group (P<0.05). TNF-α, IL-8, IL-1, Cr and BUN were significantly lower in PHC1 (P<0.05) than in the PHC2 and PHC3 groups, and ICAM-1 and NF-κB were similar between 3 PHC groups (P>0.05). Compared with the model group, the 3 PHC groups showed lessened pathological changes in the renal tubules.

CONCLUSIONPHC has protective effects against renal injury induced by hemorrhagic-endotoxin shock in rats, and treatment with 1 mg/kg PHC produces the most significant protective effect.

Acute Kidney Injury ; drug therapy ; etiology ; Animals ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1 ; blood ; Interleukin-8 ; blood ; Kidney ; drug effects ; metabolism ; Kidney Tubules ; drug effects ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; NF-kappa B ; metabolism ; Quinuclidines ; pharmacology ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; blood ; metabolism ; Tumor Necrosis Factor-alpha ; blood