Objective To analyze the genetic diversity of eight species of Caragana Fabr. in Ordos Plateau. Methods To study the genetic diversity of the eight species with ISSR method. Results The total percentage of polymorphic bands (PPB) of the eight species was up to 100%, which was extremely high and showed an abundant genetic diversity. However, the genetic diversity of different species was not the same. The genetic diversity of C. tibetica, C. korshinskii, C. stenophylla, C. roborovskyi, and C. intermedia was higher, while that of C. opulens and C. purdomii was lower, and that of C. brachypoda was the lowest. The analytic results of the Nei′s gene diversities and the Shannon′s information index were similar. Different species could be differentiated through UPGMA method and characteristic banding pattern of ISSR. Conclusion ISSR Molecular marker could be used in molecular authentication and hereditary background study of the species of Caragana Fabr. These results provide the scientific basis of developing effective protection measures for these species.

In order to analyze the correlation between critical residues in the catalytic centre of BSH and the enzyme substrate specificity, seven mutants of Lactobacillus salivarius bile salt hydrolase (BSH1) were constructed by using the Escherichia coli pET-20b(+) gene expression system, rational design and site-directed mutagenesis. These BSH1 mutants exhibited different hydrolytic activities against various conjugated bile salts through substrate specificities comparison. Among the residues being tested, Cys2 and Thr264 were deduced as key sites for BSH1 to catalyze taurocholic acid and glycocholic acid, respectively. Moreover, Cys2 and Thr264 were important for keeping the catalytic activity of BSH1. The high conservative Cys2 was not the only active site, other mutant amino acid sites were possibly involved in substrate binding. These mutant residues might influence the space and shape of the substrate-binding pockets or the channel size for substrate passing through and entering active site of BSH1, thus, the hydrolytic activity of BSH1 was changed to different conjugated bile salt.
Amidohydrolases ; genetics ; metabolism ; Bile Acids and Salts ; metabolism ; Escherichia coli ; metabolism ; Gene Expression ; Lactobacillus ; enzymology ; genetics ; Substrate Specificity

OBJECTIVETo evaluate the effect of length of tibial stump on proprioceptive recovery after anterior cruciate ligament (ACL) reconstruction.

METHODSFrom March 2011 to December 2011,42 patients with ACL tear were performed with reconstruction. The patients were divided randomly into three groups: group A, the patients with remained length of tibial stump ranging from 15 to 20 mm, including 8 males and 6 females, with an average age of (28.2 ± 6.6) years old; group B, the patients with remained length of tibial stump ranging from 5 to 10 mm, including 9 males and 5 females, with an average age of (27.9 ± 6.4) years old; group C, the patients with remained length of tibial stump less than 2 mm, including 9 males and 5 females, with an average age of (28.6 ± 6.8) years, old. The stability of knee were assessed by Lanchman test and anterior drawer test. The function of knee was assessed by Lysholm score and Tegner activity rating. The proprioceptive recovery was evaluated by assessing the passive reproduction of the angles with limb movement into flexion and extension in no weight bearing position.

RESULTSLanchman test and anterior drawer test of all patients were negative at 6 months postoperatively., and there was one case of positive outcome in each group at 12 month and 18 month postoperatively. Lysholm score and Tegner activity score of all patients at 18 month postoperatively were significantly better than that preoperatively, and there were no significant difference among three groups at 18 month postoperatively. There were no significant difference in the production of the angles at flexion to 20° and extention to 80° and 20° among these groups before and after operation. The reproduction of the angles of group A and B at flexion to 50° and extention to 50° at 6 month postoperatively were significantly better than that of group A and B preoperatively and that of group C at 6 month postoperatively, and there were no significant difference among three groups at 18 month postoperatively. The reproduction of the angles of group A and B at flexion to 80° at 12 month postoperatively were significantly better than that of group A and B preoperatively and that of group C at 12 month postoperatively, and there were no significant difference among three groups at 18 month postoperatively.

CONCLUSIONPreservation of tibial stump in ACL reconstruction has benefit in proprioceptive recovery at early stage postoperatively and the length of tibial stump should be reserved with a range from 5 to 10 mm.

Adult ; Anterior Cruciate Ligament ; physiopathology ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Female ; Humans ; Knee Joint ; physiopathology ; surgery ; Male ; Proprioception ; Range of Motion, Articular ; Tibia ; chemistry ; physiopathology ; surgery ; Young Adult

Objective:To investigate the effect of poly (lactic acid)(PLA)electrospun membranes loaded with cisplatin and chloroquine on the oral squamous cell carcinoma CAL-27 cells,and to explore the method to prevent the recurrence of oral cancer.Methods: The DDP/PLA membranes, CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were prepared by electrospinning.Then the micro morphology of three kinds of membranes were observed by scanning electron microscope (SEM);the degradation rate of PLA membrane was measuredby UV spectrophotometric.The LC3-Ⅱ expression level in CAL-27 cells was detected by laser scanning confocal microscope.The survival rate of CAL-27 cells was detected by MTT method.Results:The SEM results showed that the nanofibers of DDP/PLA,CQ/DDP/PLA and CQ/DDP/CQ/PLA membranes were continuous and smooth with uniform diameters.The degrated time of membranes was about 21 d.The MTT result showed that compared with control group,at first,the effects of cell killing of DDP/PLA membranes,CQ/DDP/CQ/PLA membranes and CQ/DDP/PLA membranes were not obvious;as the extension of time,the survival rates of CAL-27 cells in DDP/PLA membranes group,CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were decreased (P <0.05).The immunofluorescence results showed that the fluorescence intensity of LC3-Ⅱ in CQ/DDP/CQ/PLA membranes group and CQ/DDP/PLA membranes group were lower than that in DDP/PLA membranes group.Conclusion:CQ/DDP/PLA membranes with sustained-release effect can increase the sensitivity of CAL-27 cells to DDP and enchance the killer effect of DDP on the CAL-27 cells.

AIM: To investigate the effects of sodium ferulate on cholesterol and triglyceride metabolism in atherosclerosis with hyperlipidemia.METHODS: The rabbit model of atherosclerosis was produced by feeding high lipid forages.RAW264.7 foam cell and HepG2 injured cell models were established by incubation with oxidized low density lipoprotein(ox-LDL).The atherosclerotic plaque area was measured,and serum lipids were detected.The cellular lipid accumulation was examined by oil red O staining.The cellular contents of total cholesterol and cholesterol ester were quantified by high performance liquid chromatography.The hepatic lipase(HL) mRNA expression was determined by RT-PCR.RESULTS:(1) Compared with hyperlipid group,the aorta atherosclerosis plaque area and the serum triglyceride level were significantly decreased in sodium ferulate-treated rabbits,but the serum cholesterol level showed little change.(2) Compared with ox-LDL group,the HL mRNA expression in HepG2 cells was enhanced significantly in sodium ferulate-treated group,but the cellular contents of total cholesterol and cholesterol ester in RAW264.7 foam cells showed little change.CONCLUSION: Sodium ferulate inhibits the formation of atherosclerotic plaque in high-cholesterol-fed rabbits aorta.This antiatherosclerotic function may reduce serum triglyceride level through enhancing the expression of HL mRNA without influencing serum cholesterol level and foam cell formation.

Pregnane X receptor (PXR), a member of nuclear receptor superfamily, plays an important role in xenobiotic and endogenous metabolism, endocrine balance, and cell proliferation, etc. Previous study has shown that pregnenolone 16α-carbonitrile (PCN), a mouse PXR agonist, could induce liver enlargement. And we found that the change in hepatocytes exhibits regional distribution characteristics: hepatocyte enlargement occurs around the central vein (CV) area, while hepatocyte proliferation occurs around the portal vein (PV) area. In this study, the dynamic changes of hepatocytes during PXR-induced liver enlargement were determined. Serum and liver samples from male C57BL/6 mice were collected for biochemical and pathological analysis after PCN treatment for 1, 2, 3, 5 days, respectively. The animal experiment was approved by the Animal Ethics Committee of Sun Yat-Sen University. The results showed that with the increase in the PCN treatment days, the feature of this regional change of hepatocyte around the CV and PV areas became more and more obvious. At the same time, the factors related to hepatocyte enlargement, such as the expression of PXR downstream genes and the hepatic content of triglyceride (TG), has gradually increased. The upregulation of proliferation-related proteins and downregulation of cyclin-dependent kinases inhibitor proteins were observed in the early stage of PCN treatment, suggesting that hepatocyte proliferation occurs earlier than hepatocyte enlargement during PXR-induced liver enlargement. This study reveals the dynamic change of hepatocytes during PXR-induced liver enlargement and provides a new insight in liver enlargement promoted via PXR activation.

Objective:To analyze the expression of LETM2 in KYSE150 and ECA109 cell lines and its effect on the proliferation, migra-tion, and invasion of esophageal squamous cell carcinoma (ESCC). Methods:The expression level of the LETM2 protein in 90 paired hu-man ESCC tissues and matched adjacent normal tissues was determined through immunohistochemistry. The expression level of LETM2 in ESCC cell lines was detected by real-time PCR and Western blot. The expression levels of LETM2 in KYSE150 and ECA109 cell lines were knocked down using lentivirus. MTT assays were performed to examine the effect of LETM2 on the proliferation of ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle. The effect of LETM2 depletion on the migration and invasion of ESCC cells was determined by Transwell assay. Results:LETM2 expres-sion was frequently upregulated in the ESCC tissues than in the adjacent normal tissues. The suppressed exogenous expression of LETM2 led to the inhibition of cell proliferation and colony formation. However, cell migration and invasion were not affected. The re-sults on the cell cycle distribution revealed that LETM2 knockdown acts as a negative regulator of the cell cycle at the G1 to S phase transition. Conclusion:LETM2 acts as a tumor-driven gene in the development and progression of ESCC. This finding suggests that LETM2 can be used as an efficient prognosis biomarker and a potential therapeutic target for ESCC.

OBJECTIVETo investigate the role of pulp in the root resorption of primary teeth without permanent tooth germs.

METHODSThe animal model without permanent tooth germs was established by surgery in Beagle dog. The root resorption was observed by taking periapical radiographs periodically. The samples of mandibular bone and pulp at different resorption stages were collected. The distribution of odontoclasts and the activating factor was analyzed by histological staining and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The role of pulp in the root resorption of primary teeth was tested by early pulpectomy.

RESULTSIn the root resorption of primary molars without permanent teeth germs, a large number of odontoclasts were present on the pulpal surface of the root canal. Semi-quantification RT-PCR showed that the ratios of the expression of receptor activator of NF-κB ligand (RANKL) mRNA and β-actin in the pulp of permanent teeth and primary teeth without permanent teeth germ during different periods of root resorption are 0.1314, 0.1901, 0.2111 and 0.6058 (P > 0.05). The root resorption of primary teeth without permanent teeth germs in test groups was about 5 weeks later than that of control group.

CONCLUSIONSThe pulp of primary tooth played an important role in the root resorption of primary tooth without permanent tooth germ.

Actins ; metabolism ; Animals ; Dental Pulp ; metabolism ; physiology ; Dental Pulp Cavity ; metabolism ; Dogs ; Male ; Molar ; Osteoclasts ; cytology ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Root Resorption ; metabolism ; Tooth Germ ; Tooth, Deciduous ; physiology

This study aims to construct the recombinant lentivirus vector containing specific small interfering RNA (siRNA) targeting rat CREB binding protein(CBP)gene and to identify its function of inhibiting the expressions of acetylated histone in primarily cultured hippocampal neurons. Firstly, we constructed four kinds of recombinant lentivirus siCBP. And then we used them to infect the primarily cultured hippocampal neurons, and performed real-time PCR, western blot respectively to detect the expressions of CBP. Afterwards, the most effective lentivirus siCBP was used to infect the primarily cultured hippocampal neurons, and then the HAT activity and protein expressions of acetylated histone Ac-H3, Ac-H4 of the neurons were examined. By using PCR, endonuclease cutting and gene sequencing, we confirmed that the target genes were correctly cloned in lentivirus vector. Besides, CBP mRNA and protein expressions in neurons were found to be with varying degrees of decreases after infections of the four kinds of lentivirus siCBP. Furthermore, the representative and most effective lentivirus GR806 could effectively inhibit the HAT activity and the protein expressions of Ac-H3, Ac-H4 in neurons. It provides the experimental basis for the subsequent application of siCBP to clarify the effects and corresponding molecular mechanism of the CBP-dependent histone acetylation on learning and memory function in hippocampus.
Acetylation ; Animals ; CREB-Binding Protein ; metabolism ; Genetic Vectors ; Hippocampus ; cytology ; Histones ; metabolism ; Lentivirus ; Memory ; Neurons ; metabolism ; Primary Cell Culture ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction

The aim of this study was to evaluate the influence of phosphorus (P) deficiency on the morphological and functional characteristics of erythrocytes in cows. Forty Holstein-Friesian dairy cows in mid-lactation were randomly divided into two groups of 20 each and were fed either a low-P diet (0.03% P/kg dry matter [DM]) or a control diet (0.36% P/kg DM). Red blood cell (RBC) indices results showed RBC and mean corpuscular hemoglobin decreased while mean corpuscular volume increased significantly (p < 0.05) in P-deficient cows. Erythrocyte morphology showed erythrocyte destruction in P-deficient cows. Erythrocytes' functional characteristics results showed total bilirubin and indirect bilirubin concentrations and aspartate transaminase and alanine transaminase activity levels in the serum of P-deficient cows were significantly higher than those in control diet-fed cows. Activities of superoxide dismutase and glutathione peroxidase in erythrocytes were lower, while the malondialdehyde content was greater, in P-deficient cows than in control diet-fed cows. Na⁺/K⁺-ATPase and Mg²⁺-ATPase activities were lower in P-deficient cows than in control diet-fed cows; however, Ca²⁺-ATPase activity was not significantly different. The phospholipid composition of the erythrocyte membrane changed and membrane fluidity rigidified in P-deficient cows. The results indicate that P deficiency might impair erythrocyte integrity and functional characteristics in cows.
Alanine Transaminase ; Aspartate Aminotransferases ; Bilirubin ; Diet ; Erythrocyte Indices ; Erythrocyte Membrane ; Erythrocytes ; Glutathione Peroxidase ; Malondialdehyde ; Membrane Fluidity ; Phosphorus* ; Superoxide Dismutase