ObjectiveTo investigate effects of pamidronate on the proliferation and differentiation of osteoblasts of rats in vitro.MethodsOsteoblasts isolated from newborn rat calvaria were treated with various concentrations of pamidronate, the proliferation of osteoblasts was evaluated with the method of methyl thiazole tetrazolium (MTT) and activity of alkaline phosphatase (ALP) in medium was measured with kit of ALP detecting.ResultsThe proliferation of osteoblasts increased under the stimulation of Pamidronate range 10-6-M-10-12 M(P<0.05), but was inhibited at the concentration of high level (10-4 M). The activity of ALP decreased in the experiment.ConclusionPamidronate can act on the osteoblasts directly and increase the proliferation of bone cells, but inhibit the differentiation of the same cells.

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Objective:To investigate the role of growth arrest specific protein 6 (Gas6) in regulating macrophage polarization in wound healing.Methods:Clean male B6 mice were randomly(random number) divided into the normal group, skin defect group, skin defect group + normal saline group (PBS group), skin defect + Gas6 (1 μg) group, skin defect + Gas6 (5 μg) group, and skin defect + Gas6 (10 μg) group. Ten mice in each group were used to observe the healing of skin wounds. Macrophages were isolated from the wound tissues of the remaining 6 mice on the fifth day after modeling. The levels of IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA), the mRNA expression levels of arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) were detected by RT-PCR, and flow cytometry was used to detect the expression of M1 marker CD197, M2 marker CD163 and F4/80. HE staining was used to detect the pathological changes of skin wounds. Masson staining was used to analyze the granulation tissue and collagen deposition.Results:Scab began to form on the surface of the wound on the third day after the skin defect model was established. The wound area of the Gas6 treatment group was smaller than that of the PBS group, and the wound healing was better than that of the PBS group. Compared with the normal group, the proportion of CD197 in macrophages of the skin defect group was significantly increased ( P=0.00 49), the proportion of CD163 and F4/80 double positive was significantly decreased ( P=0.00 86), the level of IL-6 was significantly increased ( P=0.00 13), the level of IL-10 was significantly increased ( P=0.00 14), the level of iNOS mRNA was significantly increased ( P=0.00 8), and Arg-1 was significantly increased in the skin defect group The mRNA level was significantly decreased ( P=0.01 21), and the inflammatory infiltration was aggravated. Compared with the PBS group, the proportion of CD197 in the Gas6 treatment group was significantly decreased ( P=0.00 0), the double positive rates of CD163 and F4/80 were significantly increased ( P = 0.00 0), the level of IL-6 was significantly decreased (P = 0.00 0), the level of IL-10 was significantly increased ( P=0.00 03), the level of iNOS mRNA was significantly decreased ( P=0.00 18), the level of Arg-1 mRNA was significantly increased ( P=0.00 1), and the number of inflammatory cells and the number of collagen fibers were increased. Conclusions:Gas6 can promote the transformation of macrophages from M1 to M2 in mice with skin defect, which is beneficial to the wound healing of skin defect.

Objective: To study the effect of berberine (BR) on ventricular remodeling in experimental rats with myocardial infarction (MI) and its mechanisms.
Methods: The MI model of experimental rats was established by ligation of the left anterior descending coronary artery and the MI animals were randomly divided into 3 groups: MI+BR group, in which the rats received BR 20 mg/kg.d, Sham group and MI group, the rats in those 2 groups received the same volume of normal saline. All animals were treated for 8 weeks. The cardiac function and structure were assessed by echocardiography, cardiac interstitial collagen deposition was evaluated by Masson stain, the myocardial cell apoptosis was detected by Tunel method, and the activation of nuclear factor (NF-κB) was also examined.
Results: For echocardiography, MI group had enlarged left ventricular end diastolic diameter (7.28 ± 0.29) mm than Sham group (6.86 ± 0.36) mm,P<0.05, but it decreased in MI+BR group (6.89 ± 0.99) mm,P>0.05. MI group had increased left ventricular end systolic diameter (5.88 ± 0.33) mm than Sham group (4.61 ± 0.31) mm, but it decreased in MI+BR group (4.68 ± 1.17) mm, allP< 0.01. MI group showed increased left ventricular posterior wall compensatory hypertrophy (1.81 ± 0.85) mm than Sham group (1.67 ± 0.16 mm),P<0.05, while in MI+BR group, it was deereased to (1.65 ± 0.14) mm. MI group presented decreased LVEF (45.77 ± 3.17) % than Sham group (67.28 ± 4.15) %, but it increased in MI+BR group (64.64 ± 5.82) %, allP<0.01. For Masson stain, cardiac interstitial collagen deposition in MI group (11.39 ± 0.45) % was higher than Sham group (2.65 ± 0.45) %, but less in MI+BR group (7.00 ± 0.87) %, allP<0.01. For Tunel examination, the myocardial cell apoptosis index was increased in MI group (21.31 ± 2.34) than Sham group (0.99 ± 0.38), but decreased in MI+BR group (14.15 ± 1.62), allP<0.01. For NF-κB activation study, the nuclear protein p65 content was higher in MI group (0.14 ± 0.02) ng/ml than Sham group (0.06 ± 0.01) ng/ml, but lower in MI+BR group (0.10 ± 0.02) ng/ml, allP<0.01.
Conclusion: Application of BR may improve the ventricular remodeling and cardiac function in experimental MI rats, it might be because of BR partially inhibit NF-κB activation, reduce collagen deposition and help anti-apoptosis in myocardial cells.

PurposeTo investigate the clinical value of inverted X-ray plain film and MRI examination in the diagnosis of congenital anorectal malformation (CARM). Materials and Methods Thirty-eight cases with operatively proved anorectal malformation were reviewed; inverted X-ray plain film and MRI examination were performed in all patients before surgery. The relationship between the rectum blind side and pubococcygeal line (PC line), and the type of anal atresia was determined, to compare the diagnostic accuracy of inverted plain film with MRI for CARM typing. Results Of all the 38 cases, 19 cases were with low imperforate anus, 8 cases with median imperforate anus, and 11 cases with high imperforate anus. The accuracy rate of inverted X-ray plain film and MRI examination for the diagnosis of CARM typing was 92.1% (35/38) and 97.4% (37/38) respectively, and the difference between them was not statistically significant (χ2=1.37, P>0.05). 7 cases of fistula, 5 cases of spinal cord malformations and 1 case of right kidney agenesis can be clearly demonstrated on MRI. Conclusion Both inverted X-ray plain film and MRI can diagnose the typing of CARM accurately, but MRI is also able to diagnose the fistula, visceral, spinal cord lesion and other abnormalities accompanied with CARM, while reducing the dose of X-ray radiation and damage in children, thus has higher clinical application value compared with inverted X-ray plain film.

Objective To study whether Microtus fortis can be infected with schistosome in wild. Methods Two villages (Banghu Village of Yueyang County and Nangang Village of Yuanjiang City) were selected as the study pilots. M. fortis were captured from both outside and inside embankment of the 2 villages. The liver, portal vein and mesentery vein of the captured M. fortis were examined for schistosome eggs, adult worms and schistosomula. Results A total of 1 440 M. fortis were captured, and after examined there were no eggs, adult worms and schistosomula of schistosome found. Conclusion M. fortis can not be infected with schistosome in wild environment.

To determine the genetic diversity of Haloxylon ammodendron collected from 14 sites in 5 provinces, 103 H. ammodendron samples of 12 wild populations and 2 cultivated which collected from 14 sites in 5 provinces were analyzed by amplified fragment length polymorphism (AFLP) DNA markers. PopGen32 and NTSYSpc2.1 was applied to evaluate genetic diversity of H. ammodendron populations. The average percentage of polymorphic loci (PPL) of total H. ammodendron populations was 94.13%, the average Nei's gene diversity index (H(e)) from 14 populations was 0.308 0, and the Shannon's genetic diversity index (I) was 0.467 6. The results indicated that the genetic diversity of H. ammodendron populations was high. Genetic differentiation index (G(st)) was 0.313 8, and the gene flow (N(m)) was 1.093 5 at the population level. The level of gene flow of H. ammodendron showed it possessed the feature of wind-pollinated outcrossing plants. AMOVA analysis indicated that genetic variation of H. ammodendron was much higher within groups (89.34%) than that among groups (10.66%), moreover genetic variation within groups mainly occurred among populations in different producing areas (84.80%). Cluster analysis (UPGMA) was applied to generate dendrogram based on Nei's genetic distances of 14 populations. Samples from Xinjiang and Qinghai were clustered respectively as a clade for their distant genetic relationship, while Samples from Gansu, Inner Mongolia and Ningxia were clustered together for their close genetic relationship. Genetic diversity of H. ammodendron populations is high in China, and genetic differentiation among regions is small, thus abundance within this specie is high at this stage. Therefore, wild nursery and artificial cultivating in different areas are effective measures for the conservation and sustainable utilization of H. ammodendron resources.
Amaranthaceae ; genetics ; Amplified Fragment Length Polymorphism Analysis ; China ; Evolution, Molecular ; Genetic Variation ; Phylogeny

To study the B cell linear epitopes of rabies virus CVS-11 nucleoprotein, peptides were synthesized according to the amino acid sequences of B cell linear epitopes. Linear epitopes predicted by bioinformatics analysis were evaluated with immunological techniques. Indirect enzyme-linked immunosorbent assay showed that titers of antibodies to peptides (355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein) were above 1:12 800 in mouse sera. The antibodies recognized denatured rabies virus CVS-11 nucleoprotein in Western blot analysis. Purified anti-peptide antibodies recognized natural rabies virus CVS-11 nucleoprotein in BHK-21 cells in indirect fluorescent antibody test. The 355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein were validated as B cell linear epitopes.
Amino Acid Sequence ; Animals ; Antibodies, Viral ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Nucleoproteins ; chemistry ; genetics ; immunology ; Rabies ; immunology ; virology ; Rabies virus ; chemistry ; genetics ; immunology

0507JS60 virus was isolated from a pool of Culex sp. collected in Kashi, Xinjiang, which could be propagated stably on C6/36 cells and caused cytopathic effects continuously. Viral particles had no envelope and appeared round with diameter of about 55nm (n = 10). Capsomeres on the surface of capsid were clearly visible. Electrophoresis of viral genome showed a profile of 12 double stranded RNA (dsRNA) segments. Sequencing of the twelfth segment revealed the length of 760bp (GenBank ID: FJ157354). A single open reading frame (ORF) was found and encoded a protein of 174 amino acids with a molecular mass of 18.9kD. The nucleotide sequence had similarity over 89% with that of LNV, but the deduced amino acid sequence had similarity over 91% with that of LNV. A phylogenetic tree was constructed to compare the corresponding genetic sequences in Seadornavirus. The tree demonstrated that 0507JS60 virus lied in the same branch with LNV and more closely related to LNV-NE9712. 0507JS60 virus was identified as LNV, which was firstly isolated outside the Northeast of China.
Animals ; Cell Line ; China ; Culex ; virology ; Molecular Sequence Data ; Phylogeny ; Reoviridae ; classification ; genetics ; isolation & purification ; ultrastructure

0507BS3 virus was isolated from a mixed pool of Culex sp. and Anopheles sp. collected in Kashi, Xinjiang, China. 0507BS3 virus could cause cytopathic effects on C6/36 cells but not on Vero and BHK-21 cells. Viral particles had no envelope and appeared round with diameter of about 60 nm (n = 20). Viral capsid was composed of a single layer and a central core. Capsomeres on the surface of capsid were clearly visible. Electrophoresis of viral genome showed a profile of 10 double stranded RNA (dsRNA) segments. Sequencing of the tenth segment revealed the length of 964bp (GenBank ID: FJ150869). A single open reading frame (ORF) was found and encoded a protein of 275 amino acids with a molecular mass of 30.8kDa. The nucleotide sequence had no similarity with any other viral genomic sequences, but the deduced amino acid sequence significantly matched the polyhedrin genes of cytoplasmic polyhedrosis virus (CPV) in some sections. A phylogenetic tree was constructed to compare the polyhedrin gene sequences of all CPV types in GenBank. The tree demonstrated that 0507BS3 virus was only distantly related to the other CPV types and belonged to a new CPV type.
Animals ; Cell Line ; China ; Culicidae ; virology ; Phylogeny ; Reoviridae ; classification ; genetics ; isolation & purification ; ultrastructure