The study aims to investigate the embryo-fetus development toxicity of the novel PPAR-δ agonist HS060098 on SD rats. The pregnant rats that were randomly divided into the solvent control group (1% hydroxypropyl methyl cellulose water solution) and HS060098 suspension groups (10, 30 and 100 mg x kg(-1) xd(-1)) were orally administered with HS060098 suspension or vehicle during the gestation of 6 -15 days (GD6-15). At termination (GD20), female rats were sacrificed. The pregnant females were evaluated by corpora lutea count, implantation sites, existence and death of embryos. Fetal sex, weight, externals, variations and malformations of viscus and skeleton were observed. The results show that there were no significant abnormality in maternal general conditions and fetal appearance as well as viscera, but in the 100 mg x kg(-1) x d(-1) group, the maternal weight gain decreased greatly (P < 0.01) and the skeletal ossification delayed remarkably (P < 0.01); in the 30 mg x kg(-1) xd(-1) group, the fatal and litter number of incompletely ossified sternebrae II was higher than those of the control group (P < 0.05); the skeletal malformations occurred in all dose groups, which indicate that the novel PPAR-δ agonist HS060098 had maternal toxicity and adversely effected fetal skeletal development under the experimental conditions.
Animals ; Bone and Bones ; drug effects ; Embryonic Development ; drug effects ; Female ; Fetal Weight ; PPAR delta ; agonists ; Pregnancy ; Rats ; Toxicity Tests

OBJECTIVETo determine the molecular basis of late onset ornithine transcarbamylase (OTC) deficiency in a Chinese family of Han nationality and the exon sequences of OTC gene of this patient.

METHODSPolymerase chain reaction-single strand conformation polymorphism and direct sequencing were used to identify the mutation type.

RESULTSA missense mutation E122G in the conserved residue of exon 4 was identified which is unreported before.

CONCLUSIONThe E122G mutation in human OTC gene may cause late onset OTC deficiency.

Age of Onset ; Base Sequence ; Child, Preschool ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Fatal Outcome ; Female ; Humans ; Male ; Models, Molecular ; Mutation, Missense ; Ornithine Carbamoyltransferase ; chemistry ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; enzymology ; genetics ; pathology ; Pedigree ; Polymorphism, Single-Stranded Conformational ; Protein Structure, Secondary

OBJECTIVETo analyze the clinical and cytogenetic features of myelodysplastic syndrome(MDS) associated with del(20q).

METHODSThe cytogenetic profiles, clinical manifestations, laboratory data, and transformation in course of disease were analyzed.

RESULTS(1) Of 29 MDS patients with del(20q), eleven (37.9%) had normal karyotype in addition to del(20q) aberration. Among them, nine patients were categorized into refractory anemia(RA)/RA with ringed sideroblasts(RAS) group and two into RA with excess Hasts(RAEB)/RAEB in transformation(RAEB-T) group. The breakpoint in 20q11 was commonly seen in patients with RA/RAS(63.2%), while del(20q12) was predominant in patients with RAEB/RAEB-T(accounting for 70% in all RAEB/RAEB-T patients). It was observed that RAEB/RAEB-T patients had higher frequencies of extra chromosomal aberrations(50%) and complex karyotype(30%) than did the RA/RAS patients (26.3%, 5.3% respectively); (2) Almost all patients revealed prominent pancytopenia, dyserythropoiesis and dysgranulopoiesis and 58.6% patients showed dysmegakaryopoiesis; positive periodic acid schiff staining of nucleated erythrocytes or reduction of neutrophils were found in 62.5% of patients; 81.8% of patients expressed lymphoid antigens; (3) Two cases transformed to acute myeloid leukemia.

CONCLUSIONDel(20q) may be an early and primary cytogenetic event in the development of hematologic malignancies. Pancytopenia and dysplasia of bone marrow cells are prominent in patients with MDS associated with del(20q); lymphoid antigen expression is a common occurrence; more additional chromosomal abnormalities and complex karyotypes appear when the disease becomes worse.

Adolescent ; Adult ; Aged ; Child ; Chromosome Deletion ; Chromosomes, Human, Pair 20 ; Female ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology

OBJECTIVETo investigate the effects of hydrogen sulfide (H2S) on H2O2-stimulated primary neonatal rat cardiomyocytes and related mechanism.

METHODSPrimary neonatal rat cardiomyocytes were treated with various concentrations of H2O2 (10, 100, 1000 µmol/L) for 24 h to establish the oxidative stress-induced cell injury model after 3 days' conventional culture. In addition, different concentrations of NaHS (1, 10, 100 µmol/L) were added to cardiomyocytes in the absence and presence of 100 µmol/L H2O2 for 24 h. The viability of cardiomyocytes was measured by MTT assay. The SOD vitality was measured by xanthine oxidase method and MDA content was determined by thiobarbituric acid colorimetric method. LDH activity was measured by chemical colorimetric method. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 (Rh123) staining and photofluorography. The level of reactive oxygen species (ROS) in cardiomyocytes was measured by DCFH-DA staining and photofluorography.

RESULTSCell viability and SOD vitality were significantly reduced while MDA content and LDH activity were significantly increased with increasing H2O2 concentrations. These effects could be partly reduced by cotreatment with H2O2 in a concentration-dependent manner (all P < 0.05). Compared with control group, the DCF fluorescence intensity significantly increased in the 100 µmol/L H2O2 group (P = 0.003), which could be attenuated by NaHS in a dose-dependent manner. Compared with control group, the MMP significantly decreased in the 100 µmol/L H2O2 group (P = 0.000), which could be partly reversed by cotreatment with NaHS in a dose-dependent manner. Moreover, H2O2 treatment also significantly reduced 100 µmol/L H2O2 induced apoptosis in a dose-dependent manner.

CONCLUSIONH2S protects primary neonatal rat cardiomyocytes against H2O2-induced oxidative stress injury through inhibition of H2O2 induced overproduction of ROS, dissipation of MMP and apoptosis.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Hydrogen Peroxide ; pharmacology ; Hydrogen Sulfide ; pharmacology ; Malondialdehyde ; metabolism ; Membrane Potential, Mitochondrial ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Superoxide Dismutase ; metabolism

OBJECTIVETo ascertain an accurate approach to inserting the pedicle screw into C3-C7 segments of the cervical vertebra.

METHODSAnatomic morphology of lateral mass and pedicle, and their anatomic relationship with the adjacent tissue were observed on C3-C7 segments of 25 adult embalmed cadavers (50 sides).

RESULTS1) The inferior edge of the base of the posterior tubercle of the transverse process and the inferior edge of the pedicle were connected with each other on 25 adult embalmed cadavers (50 sides). The transverse section which passed through the median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, and the transverse section which passed through the central axis between the superior edge and the inferior edge of the pedicle, were in the same horizontal plane. The superior and inferior position of placing the pedicle screw was determined by this transverse section, which passed through the median point between the superior and the inferior edge of the base of the posterior tubercle of the transverse process. 2) There was a directed internal-downwards "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process. The anterior wall of the triangular sulcule was the base of the posterior tubercle of the transverse process, the posterior wall was the anterolateral edge of the inferior articular process, and the bottom of the sulcule was connected with the interior edge of the pedicle. The vertical length between the top of triangle and the planes of inferior edge of the pedicle was (2.78+/-1.71) mm. The inferior edge of the cervical pedicle could be detected using a blunt probe along the "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process in surgical operation. 3) The lateral fovea of the articular process was observed on all lateral masses (50 sides). The internal and external position of the entrance point could depend on anatomic landmarks: the lateral edge of the lateral fovea of the articular process. The horizontal length between the lateral fovea of the articular process and the entrance point was (3.14+/-1.45) mm. 4) The diameter of pedicle screw, about (2.78+/-1.71) mm, was the transverse diameter of the cancellous bone of the greatest narrow part of the cervical pedicle.

CONCLUSIONSThe median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, the lateral fovea of the articular process, and the triangular sulcule between the base of the posterior tubercle of the transverse process and the anterolateral edge of inferior articular process, are easy to be exposed and identified in surgical operation. The pedicle screw can be precisely inserted through this method.

Adult ; Bone Screws ; Cervical Vertebrae ; anatomy & histology ; surgery ; Humans

OBJECTIVETo explore risk factors and infection characteristics of secondary pancreatic infection in severe acute pancreatitis (SAP).

METHODSA clinical data of 49 patients with secondary pancreatic infection in severe acute pancreatitis (SPI group)were matched with 49 patients without infection in severe acute pancreatitis (NSPI group) between January 2003 and December 2005. The two groups were analyzed by a case-control study. Conditional Logistic regression model univariate and multivariate were used to screen out risk factors. The types of infection, the peak infection and the bacteria spectrum were analyzed in SPI group.

RESULTS(1) In univariate Logistic regression analysis, 7 factors including continuous hypoalbuminemia, prolonged time of central venous catheter, usage of hormone, high APACHE II scores, multi-antibiotics, intestine dysfunction and continuous hyperglycemia were selected out. Moreover, the first three were statistically significant in multivariate Logistic regression analysis. (2) Pancreatic abscess ranked first in SPI group. Of all the pancreatic infection, 22.5% occurred within two weeks and 71.4% occurred in the 4th week or later. (3) In SPI group, 81 strains of microorganisms were cultured, including 45 strains of gram-negative bacteria (55.6%), 22 strains of gram-positive bacteria (27.2%), and 14 strains of fungi (17.3%). The common gram-negative bacteria were Escherichia coli, and the common gram-positive bacteria were Staphylococci and Enterococci. The fungi included Monilia and Yeastoid fungus. Further study revealed that 35 strains of all the microorganisms were intestinal bacteria (43.2%).

CONCLUSIONSContinuous hypoalbuminemia, prolonged time of central venous catheter and usage of hormone were independent risk factors of SPI. The main type of infection was pancreatic abscess. Gram-negative bacteria, were the common bacteria causing secondary pancreatic infection.

Adult ; Aged ; Bacterial Infections ; etiology ; microbiology ; Case-Control Studies ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Pancreatitis ; etiology ; microbiology ; Pancreatitis, Acute Necrotizing ; complications ; Retrospective Studies ; Risk Factors

OBJECTIVETo study the effect of Itk down regulation on Jurkat cell proliferation and inflammatory cytokines production, and provide useful data for Itk as an attractive target for potential drugs.

METHODSThree shRNAs against different region of Itk were constructed and cotransfected with pEGFP-C1-hItk. The shRNA, which can knock down Itk, was selected and packed into lentivirus. After Jurkat cells were transfected with shRNA lentivirus, the change of Itk protein expression, cell proliferation and cytokines production was observed.

RESULTSItk mRNA was reduced about 55% in Jurkat cells transfected with Itk-shRNA1, compared with that in control cells shRNAnon (P < 0.05). Knocking down Itk expression had a profound inhibitory effect on Jurkat cell proliferation. In addition, there was a substantial decrease in level of cytokines, such as IL-2, IL-5, IL-10 and IFN-gamma, produced by cell transfected with Itk-shRNA1.

CONCLUSIONKnocking down Itk expression can inhibit Jurkat cell proliferation and inflammatory cytokines production.

Animals ; Cell Proliferation ; Cytokines ; genetics ; immunology ; Down-Regulation ; Gene Knockdown Techniques ; Humans ; Interleukin-10 ; genetics ; immunology ; Interleukin-2 ; genetics ; immunology ; Interleukin-5 ; genetics ; immunology ; Jurkat Cells ; cytology ; enzymology ; immunology ; Mice ; Protein-Tyrosine Kinases ; genetics ; immunology

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; complications ; pathology ; physiopathology ; Liver ; pathology ; physiopathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the association of gene polymorphisms of Toll-like receptor 3 (TLR3)-1377C/T and expression of TLR3 with the susceptibility to enterovirus 71 (EV71) encephalitis in children.

METHODSA total of 187 children with EV71 infection (59 children in the encephalitis group and 128 in the non-encephalitis group) and 232 children who underwent physical examination were enrolled in the case-control study. Polymerase chain reaction-restriction fragment length polymorphism was used to detect the TLR3-1377C/T gene polymorphisms. ELISA was used to measure the serum level of TLR3.

RESULTSThere were no significant differences in the genotype and allele frequencies of TLR3-1377C/T between the non-encephalitis group and the encephalitis group. Compared with the control group, the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 (P<0.05), and the non-encephalitis group had the highest level (P<0.05). The encephalitis group had a significantly higher EV71 viral load than the non-encephalitis group (P<0.01). The children aged <1 year or ≥1 year in the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 compared with their counterparts in the control group (P<0.05), and the children aged <1 year or ≥1 year in the non-encephalitis group had a significantly higher serum level of TLR3 than those in the encephalitis group (P<0.05). In the encephalitis group, the children aged ≥1 year had a significantly higher TLR3 concentration than those aged <1 year (P<0.05), and there were no significant differences in the TLR3 concentration between the children aged ≥1 year and <1 year in the non-encephalitis group and the control group. In the encephalitis group, the proportion of children aged <1 year was significantly higher than those aged ≥1 year (P<0.05).

CONCLUSIONSThe TLR3-1377C/T gene polymorphisms are not significantly associated with the development of EV71 encephalitis. Low expression of TLR3 might weaken the inhibitory effect on virus replication and promote the development of EV71 encephalitis. The deficiency in the expression of TLR3 in serum after EV71 infection might be an important factor for the development of encephalitis in infants.

Child, Preschool ; Encephalitis, Viral ; genetics ; Enterovirus A, Human ; Enterovirus Infections ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Infant ; Male ; Polymorphism, Single Nucleotide ; Toll-Like Receptor 3 ; genetics

BACKGROUND: Previous studies have found that polygonatum sibiricum polysaccharide (PSP) exhibits anti-osteoporosis effect, but its therapeutic effect in ovariectomized osteoporotic rats and the molecular mechanisms are poorly understood. OBJECTIVE: To investigate the effect of administration of PSP on the bone microstructure, bone mineral density as well as osteoblast- and osteoclast-related gene expression in rats. METHODS: Twenty-five infertile female Sprague-Dawley rats aged 3 months were randomly allotted into five groups (n=5 per group): sham operation (same volume normal saline), model, zoledronate (0.2 mg/kg?d), high-dose PSP (800 mg/kg?d) and medium-dose PSP (400 mg/kg?d) groups. All rats were subjected to ovariectomy except sham operation group. The administration was intragastrically given every 2 days beginning at 7 days after modeling and lasted 12 weeks. Then, the rats were sacrificed, and the uterus was weighed. The bilateral tibias were removed, one side for histomorphometric analysis by micro-CT, and the other one for RNA detection by qualified PCR. RESULTS AND CONCLUSION: Compared with the sham operation group, the rat body mass in the model group was significantly increased and the weight of uterus was significantly decreased (P < 0.05). Compared with the model group, zoledronate and high-dose PSP could significantly alleviate the excessive increase in body mass (P < 0.05). The bone mineral density in the model group was decreased by 63% compared with the sham operation group (P < 0.01), Compared with the model group, after 12-week high-dose PSP and zoledronate administration, the bone mineral density was increased by 44% and 38%, respectively (P < 0.01); the trabecular bone volume fraction and trabecular number rose significantly(P<0.05),while the trabecular separation decreased significantly(P<0.05).In vivo,PSP could significantly promote the expression levels of osteoblast-related genes (alkaline phosphatase, RUNX2, Col1a1 and osteocalcin), and significantly inhibit the expression levels of osteoblast-related genes (ACP5 and CTSK) (P < 0.05). These results imply that high-dose PSP can reduce bone loss and decrease of bone mineral density, improve the destruction of bone microstructure, as well as promote osteoblast-related genes but inhibit osteoclast-related gene mRNA expression in the ovariectomized rats.