Objective To determine the genetic diversity of germplasm resource in Dryopteris fragrans by amplified fragment length polymorphism(AFLP) technique.Methods Forty-six samples from six populations of D.fragrans were analyzed by AFLP DNA markers,and the genetic diversity was evaluated by NTSYS-PC 2.10 and PopGen32.Results The percentage of polymorphic bands(PPB) reached to 76.67%,observed number of alleles(Na) was 1.766 7, effective number of alleles(Ne) was 1.504 6,Nei′s gene diversity index(H) was 0.292 1,Shannon information index(I) was 0.431 6,and genetic differentiation index(Gst) was 0.412 5.Conclusion Genetic diversity is high,gene diversity in populations(Hs) is 0.170 5,and gene diversity among populations(Dst) is 0.119 8.Because of environmental specificity,its resource on the spot should be protected.

The procedures for the calculation and measurement and calibration of output dose of the Siemens primus E accelerator with IAEA regulation are introduced to probe an accurate,rapid and effective method.The parameter value of measurement is determined according to IAEA regulations and absorbed dose calculation formal is briefed to the form that the reading of instrument multiplies exposure calibration coefficient Nx and Air temperature,pressure and humidity effects Ktp and cumulative correction factor Cf the precision of accelerator conform to the standard of QA&QC by the measurement and calibration of the Hospital Primus E accelerator in output dose.The methods of measurement and calibration can be used for the determination of output dose in precise radiotherapy of digital medical linear accelerator and clinical treatment.

Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.

Objective To explore the therapeutic effect and possible mechanisms of Xuebijing injection for treatment of acute pulmonary thromboembolism(APE)in rabbits. Methods Twenty-four New Zealand rabbits with big ears were randomly divided into control group,model group and Xuebijing group(each n=8). The model of APE was performed by re-infusing of autologous blood clots. After the model was established,in the rabbits of the Xuebijing group,Xuebijing injection 1 mL/kg was injected into the ear marginal vein,twice daily for consecutive 7 days. In the model and control groups,the same amount of normal saline was given via the same route. The changes in plasma levels of B-type natriuretic peptide(BNP)were detected by enzyme-linked immunosorbent assay(ELISA) at the time points of 8 hours and 1,3,5,7 days,and after 7 days the lungs of the rabbits were removed for the lung histopathological examination. Results The BNP levels of the model group and Xuebijing group were significantly higher compared with the level of control group at 8 hours after embolization,the model group being the most significant(all P<0.05). On day 3,the BNP levels of the model group and Xuebijing group reached their peaks,and then gradually decreased,but the level of model group was significantly higher than that of Xuebijing group(ng/L:581.1±144.6 vs. 251.5±22.5,P<0.05). On day 7,the BNP level of Xuebijing group fell to the level of the control group(ng/L:78.5±9.4 vs. 50.9±11.5),the level of the model group also decreased,but it was still much higher than that of the Xuebijing group(ng/L:214.1±47.6 vs. 78.5±9.4,P<0.05). Histopathological examination showed Xuebijing could significantly reduce the infarct size with mild inflammatory cell infiltration,but the infarct size of the model group was obviously bigger than that of the Xuebijing group and accompanied by relatively more inflammatory cell infiltration. Conclusion Xuebijing injection can significantly reduce the plasma BNP level of the rabbits with pulmonary embolism,and in the mean time ameliorate the degree of inflammatory infiltration in the lung tissue and infarct.

Objective To explore the therapeutic effect and possible mechanisms of Xuebijing injection for treatment of rabbits with acute pulmonary thromboembolism (APTE). Methods Thirty-two New Zealand rabbits were randomly divided into normal control group, model group, low molecular weight heparin (LMWH) group and Xuebijing group by random number table, 8 rabbits in each group. APTE model was reproduced by re-infusing autologous blood clots, and nothing was done for the normal control group. After the model was established, intravenous injection of Xuebijing 1 mL/kg from ear marginal vein was given to the rabbits in Xuebijing group, the LMWH 100 U/kg was subcutaneously injected in LMWH group, the same amount of normal saline was given intravenously to model group and normal control group, twice daily for 7 days in all the groups. The changes of plasma D-dimer and endothelin-1 (ET-1) levels were detected by enzyme-linked immunosorbent assay (ELISA) after modeling for 8 hours and 1, 3, 5, 7 days;after 7 days the animals were sacrificed and their lung tissues were removed for the examination of ultrastructural changes under an electron microscope. Results With the prolongation of time, the D-dimer level in the model group was gradually increased, and on the 7th day after modeling, the level was significantly higher than that of the normal control, LMWH and Xuebijing groups (g/L:3.98±1.14 vs. 0.43±0.12, 1.05±0.13, 1.04±0.15, all P<0.05). After modeling for 5 days, the ET-1 level reached its peak in the model group, and it was lowered on the 7th day after modeling, but still significantly higher than that in the normal control, LMWH and Xuebijing groups (g/L:63.8±20.9 vs. 20.0±1.4, 36.5±8.3, 34.2±6.3, all P<0.05). The D-dimer and ET-1 levels in the LMWH and Xuebijing groups were significantly higher than those of the normal control group at each time point, but still obviously lower than those of the model group (all P<0.05), and the differences between the two treatment groups were not statistically significant at each time point (all P>0.05). Electron microscopic examination of lung tissue ultra-structure showed:in the model group, there were the lung tissue basement membrane rupture, the organelle structure in vascular endothelial cells and alveolar epithelial cells typeⅠandⅡnot clear, disappearance of microvilli on cell surface, cytoplasm concentration, nuclear chromatin condensation, a lot of debris due to nuclear lysis, formation of apoptotic bodies after shedding, and mitochondrial structural damage. In LMWH group, the capillary endothelial cell swelling, a lot of necrotized blood cells oozing, blockage of lumen, the cytoplasm of typeⅡalveolar epithelial cell loose, edematous and multiple lamellar bodies vacuolized were seen. In Xuebijing group, capillary endothelial cell swelling, necrotized blood cells blocking the lumen and seeping, the link basically normal, the type Ⅱ alveolar epithelial cell edematous, mitigation of lamellar bodies emptying were found. Conclusions Xuebijing can significantly reduce the plasma D-dimer and ET-1 levels, adjust the balance of coagulation and fibrinolysis, simultaneously decrease the pulmonary vascular permeability obviously in rabbits with APTE, thus it possesses certain protective effect for endothelial function leading to amelioration of lung injury and reduction of lung pathological damage in rabbits with pulmonary embolism.

Objective To investigate the expressions and mutual relationship of Galectin-3 andβ-catenin in endome?triosis (EM). Methods Immunohistochemistry was used to detect expression levels of Galectin-3 andβ-catenin and their mutual relationship in expression was also examined. Samples were collected from ectopic endometrium of patients with en?dometriosis (ectopic endometrium group, n=34), eutopic endometrium of patients with endometriosis (eutopic endometrium group, n=34) and normal endometrium from people without endometriosis (control group, n=30). Results The expressions of Galectin-3 were seen in 88.2%, 85.3%, 50.0%of cases in ectopic endometrium group, eutopic endometrium group and control group respectively. On the other hand, the expression ofβ-catenin were seen in 55.9%, 52.9%, 26.7%of cases in ecto?pic endometrium group, eutopic endometrium group and control group respectively. In EMs patients, the expression of Galec?tin-3 andβ-catenin were significantly higher in ectopic endometrium and eutopic endometrium than those in normal endome?trium group(P<0.05). Expression of Galectin-3 was positive correlated with expression ofβ-catenin(rs=0.512, 0.428, P<0.01). Conclusion Galectin-3 andβ-catenin may play important roles synergistically in the pathogenesis of endometriosis.

Objective To study the in vitro and in vivo effects of pulsed electromagnetic fields (PEMFs) on osteoclast and osteoblast precursor cells.MethodsTo observe the in vitro effect of PEMFs,femur bone marrow cells of 8 week old female SD rats were collected.According to different treating doses,rats were divided into four treatment groups and one control group.After the treatment,the clones of granuloeyte/maerophage colony forming unit(CFU-GM) and fibroblast colony forming units(CFU-F) were measured respectively.To observe the combined in vitro and in vivo effect of PEMFs,8 week old female SD rats were randomly divided into three groups: 2-70 group,ovariectomization (OVX) group and SHAM group.Rats in the 2-70 group and OVX group were bilateral ovariectomized,while rats in the SHAM group were sham-ovariectomized.12 weeks after ovariectomization,the 2-70 group was exposed to PEMFs while the other groups were left untreated.Then,femur bone marrow cells of the rats were collected.According to the way whether the groups were treated with PEMFs,the cells were divided into six groups: 2-70 with/without treatment,OVX with/without treatment,SHAM with/without treatment.After the treatments,the clones of CFU-GM and CFU-F were measured respectively.Resultsin vitro effect of PEMFs: Compared with the control group,the CFU-GMin the treated groups reduced while the CFU-F increased.PEMFs effect in vitro and in vivo: The CFU-F in treated groupsincreased,whileno.significantdifferencesofCFU-GMwerefoundamongthegroups.Conclusion PEMFs has inhibitory effect on osteoelast precursor cells and enhances the proliferation of osteoblast precursor cells when simply applied in vitro.When PEMFs was applied in combined manner of in vitro and in vivo,it shows that PEMFs enhance the proliferation of osteoblast precursor cells but has no inhibitory effects on osteoelast precursor cells.

Objective To explore the influence of glucose-lowering rate on left ventricular function in patients with type 2 diabetes mellitus (T2DM). Methods One hundred and thirty-two cases of type 2 diabetes mellitus and 135 cases of type 2 diabetes mellitus with coronary heart disease (T2DM+CHD)received intensive glucose lowering therapy. Then, after measuring left ventricular ejection fraction (LVEF) and E/A ratio, the variation was analyzed. Results LVEF was significantly higher than that before intensive therapy in T2DMsubgroup with glucose-lowering rate less than 6 m mol · L-1 · d-1( P＜0.05 ). So was T2DM+CHD subgroup with glucose-lowering rate less than 4 mmol· L-1 · d-1 (P＜0.05). LVEF was significantly lower than that before intensive therapy in T2DM+CHD subgroup with glucose-lowering rate greater than 4 mmol · L-1 · d-1( P＜0. 05 ),while by the end of following up for 3 months, LVEF stepped up and no significant difference was observed between subgroups ( P ＞ 0. 05 ). The E/A ratio stepped up in both subgroups after intensive therapy ( P ＜ 0. 05 ).Conclusions For T2DM patients with coronary heart disease, excessively fast glucose-lowering rate may impair left ventricular function. Long-term good control of blood glucose restores the impaired left ventricular function causes by excessively fast glucose-lowering rate. After intensive therapy, left ventricular diastolic function finally improves in both subgroups regardless of the glucose-lowering rate and coronary heart disease.

Objective To analyze the diversity of salivary bacteria in patients with nonalcoholic fatty liv-er disease(NAFLD group)and healthy control group(HC group). Methods Saliva samples were collected from 24 patients with nonalcoholic fatty liver disease and 22 healthy control.Genomic DNA of the samples was extracted and extended to 16S rRNA V4-V5 hypervariable region after PCR with Illumina high-throughput sequencing tech-nology.All the data,including operational taxonomic units,diversity indexes and species annotation,etc.was ana-lyzed. Results 39118 optimized sequences in the saliva of each group were obtained,including nonalcoholic fatty liver group clustering OTUs about 308 ± 48 and healthy control group clustering OTUs about 305 ± 53.The diversi-ty analysis of NAFLD group and HC group showed that there was little difference in diversity of the whole salivary bacteria.The data of the species analysis of the saliva bacteria in two group indicate that the composition was simi-lar and the relative abundance was different in phylum and genus.In NAFLD group,the ratio of the abundance of firmicutes(20.42%)increased and the proportion of bacteroides(36.75%)decreased in phylum. Genus of the NAFLD group,the ratio of fusobacterium,porphyromonas and veillonella increased significantly,and the ratio of prevotella and[Prevotella]decreased.Conclusion In the NAFLD group,the abundance of firmicutes increased, and the abundance of the bacteria decreased.It was speculated that the imbalance of the salivary bacteria is associ-ated with nonalcoholic fatty liver disease.

Objective: To validate a pseudovirus-based neutralization assay for the detection of antibodies against human papillomavirus (HPV) in human serum samples. Methods: The specificity, accuracy, precision, range of linearity, limit of detection and robustness of the neutralization assay were evaluated using HPV-negative serum samples, vaccinated serum samples with quadri-valent vaccine, and international standards for detecting antibodies against HPV16 and HPV18. Results: Based on the data of the HPV-naïve samples, the criteria of positivity was determined as follows: the 50% inhibitory dose (ID₅₀) of the tested sample was not less than 40 and 2-fold not less than that for bovine papillomavirus. The neutralization assay showed good accuracy with a recovery rate of 87%-122% and excellent reproducibility with intra- and inter-assay variation of 5%-27% and 10%-26% respectively. The HPV16 and HPV18 international standards were used to define the limit of quantification, which was 1.28 IU/ml for HPV16 and 0.96 IU/ml for HPV18. Acceptable ranges of variation for the key parameters of this assay were defined, which showed the good robustness of the pseudovirus-based neutralization assay. Conclusion The pseudovirus-based neutralization assay for the detection of HPV antibodies showed good specificity, accuracy, sensitivity, and robustness, suggesting that it could be used to evaluate the immunogenicity of HPV vaccines.