Objective To observe the change of serum leptin in different risk stratifications of coronary heart disease (CHD) and to investigate its relationship with the severity of coronary artery lesion and the coronary artery Gensini score and its value in the coronary heart disease risk stratification .Methods According to coronary angiography ,120 research subjects were enrolled and di-vided into 4 groups :the non-CHD group ,stable angina(SAP) group ,unstable angina pectoris(UAP) group and myocardial infarc-tion group(AMI) ,respectively .The serum leptin levels in 4 groups were determined by immunoassay and the correlation between the leptin level with the coronary heart disease risk factor and biochemical markers of risk assessment was analyzed .Results The serum leptin level in the AMI group was significantly higher than that in the non-CHD group and the SAP group ,the leptin level showed the increasing trend with the increase of the coronary lesion severity and the Gensini scores and was positively related with the CHD risk stratification indicators cTnT and smoking index ,and negatively related with blood uric acid .Conclusion The serum leptin may be used as the valuable marker for evaluating the occurrence of acute coronary event and has good correlation with usual biochemical markers of CHD risk stratification and the severity of coronary artery lesion .

A method for the determination of Fe, Co, Mn and Ni in synthetic diamonds by inductively coupled plasma atomic emission spectrometry ( ICP-AES) was proposed. The synthetic diamond sample was decomposed completely, while the sample was burned in air at 1000 ℃ for 10 h, and then a mixed acid of H2 SO4 , aqua regia and HClO4 was used for the dissolving the residue of the sample. In this method, the limits of detection of Fe, Co, Mn and Ni were 0. 0147, 0. 0018, 0. 0006 and 0. 0027 mg/L, respectively. Under the optimum condition, Fe, Co, Mn and Ni in synthetic diamond sample were determined. The values of RSDs (n=7) were less than 0. 5%. The recoveries of added standard were 94. 0%-105. 0%.

A method for the simultaneous determination of oxygen and nitrogen in synthetic diamonds by inert gas high temperature extraction-impulse heating method was proposed. The sample weight, the selection of analysis power and the calibration curves of oxygen and nitrogen were discussed. Oxygen and nitrogen in analytical samples are determined. Values of RSDs (n=7) for oxygen and nitrogen were less than 4. 5% and 4. 0% respectively. The analytical results of oxygen and nitrogen obtained by the proposed method were in good agreement with those by inert gas fusion-impulse heating method.

A simple and sensitive Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) method was established to provide a new alternative for clinical diagnosis of Avian influenza A H5N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the target for amplification of nucleic acid under isothermal conditions. In current study, fifty-one experimentally infected animal specimens and viral cultures that had been tested were analyzed by RT-LAMP for NA gene and HA gene, respectively. The amplification process of LAMP was monitored in real-time by the addition of SYBR Green dye. Meanwhile, the result showed high correlation between nested PCR and RT-LAMP. The specificity of the RT-LAMP assay was confirmed by restriction enzyme digestion analysis and sequencing of the amplified product. When the sensitivity of this assay was tested by serial 10-fold dilutions of RNA molecules from specimens, it was found that the RT-LAMP method achieved theoretically a sensitivity of 10 RNA molecules. Thus, we concluded that the RT-LAMP assay has potential usefulness for rapid detection of the Avian influenza A H5N1 virus.
Animals ; Birds ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; diagnosis ; virology ; Nucleic Acid Amplification Techniques ; methods ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo observe the changes of telomere length and telomerase activity in patients with aplastic anemia (AA), and relationship with immunosuppressive therapy (IST) efficacy, to explore the pathogenesis of AA and the role of telomere length in evaluating immunosuppressive therapy efficacy.

METHODS71 cases of AA patients between September 2010 and March 2013 were enrolled into this study. 3 ml peripheral blood specimens from this cohort of patients were collected to test the telomere length in peripheral blood mononuclear cell (PBMNC) with flow-FISH and detect telomerase activity with TRAP-PCR-ELISA method.

RESULTSTelomere length and age showed negative correlation (b=-0.387, P=0.001) in normal control, NSAA and SAA + VSAA groups, telomere length became shorter with the growth of age, and normal control group telomere length decreased along with the age growth slightly greater than the other two groups (NSAA, SAA+VSAA). Besides the effect of age on telomere length, no significant difference was observed between NSAA and SAA+VSAA groups (P=0.573), and NSAA, SAA+VSAA (30.957 ± 4.502,29.510 ± 5.911)groups were significantly shorter than normal control group (51.086±10.844) (P<0.01). Telomere length in NR group (25.357±4.848)was significantly lower than normal control group (51.086 ± 10.844) (P=0.005), telomere length in CR(32.808 ± 4.685)/PR groups (30.334±4.464) compared with normal control group had no significant difference (P=0.517, P=0.254). Telomere length below 29.21% obviously decreased outcomes of IST. Telomerase activity had significant difference (χ²=20.385, P<0.01). The telomerase activity had no significant difference in terms of age and gender in three groups, multiple comparison found that telomerase activities in SAA + VSAA (0.324±0.178) (P<0.01), and NSAA (0.234±0.175) groups (P=0.002) were significantly higher than normal control group (0.107±0.083).

CONCLUSIONTelomere length of PBMNC in AA patients was significantly shortened than normal control group with telomerase activity increased, and telomere shorted more apparently in NR group, these patients should adjust the treatment as early as possible. Telomeres could predict the curative effect of IST.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Aplastic ; enzymology ; therapy ; Case-Control Studies ; Child ; Female ; Humans ; Immunosuppression ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Telomerase ; metabolism ; Telomere ; metabolism ; Treatment Outcome ; Young Adult

This study was purposed to investigate the clinical significance of the amount and activated status of T cell subsets, B cells, NK cells in peripheral blood from patients with myelodysplastic syndrome (MDS). The proportion of T cells, B cells, NK cells in peripheral blood from 30 patients with MDS and their surface activation markers of CD28, CD45RA, CD45RO, CD69, HLA-DR were analyzed by flow cytometry. Twenty-two patients were in the low risk group (RA + RAS) while eight patients were in the high risk group (RAEB + RAEBT). The result showed that the amounts of T cells (CD3+ cells) in peripheral blood from patients with MDS were lower than those in control group. The amounts of naive CD4+ cells (CD4+ CD45RA+ cells) in MDS patients were lower than those in control. The expression rates of early activation marker (CD69) and late activation marker (HLA-DR) on CD3+ cells in MDS patients were significantly higher than those in control. The abnormalities of the immunologically competent cells were mainly observed in the low risk group (RA + RAS), and were characterized by the high expression rates of CD69+ and HLA-DR+ on CD3+ cells, the decrease of B cell amounts. The amount abnormalities of T cell subsets were mainly observed in high risk group (RAEB + RAEBT), and were characterized by the decrease of CD3+ cells and CD3+ CD4+ CD8- cells (Th cells) amounts without high expression of the CD69 and HLA-DR, the decrease of NK cells amounts. It is concluded that there are the abnormalities of T cell subsets and function in the patients with MDS and may change with disease progression, so the measurement of amount and activated status of T cell subsets in peripheral blood from MDS patients can have predictive role for diagnosis of disease progression and guide of therapy.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; immunology ; Antigens, Differentiation, T-Lymphocyte ; immunology ; B-Lymphocytes ; immunology ; CD3 Complex ; immunology ; Female ; HLA-DR Antigens ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Lectins, C-Type ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; T-Lymphocyte Subsets ; immunology

The full-length genome of one human bocavirus (HBoV) and the VP1 sequences of nine HBoV were amplified from patients' samples by PCR, cloned into pGEM-T vector separately, and sequenced. In this study, the one full length gemome and nine VP1 sequences of HBoV were aligened with 14 sequences of Parvoviruses which were canonical exemplars in Parvovirinae. Phylogenetic analysis showed that HBoV capsid sequences positioned closely to B19 parvovirus, although they positioned far in phylogenetic tree based on full length genome. Many similarities were found between HBoV and B19 in capsid by alignment on secondary structural elements. Because both B19 and HBoV are the only Parvoviruses that infect mankind, so study on HBoV may be used for reference to B19 which had been studied for about 30 years. By analysis of mutational sites, HBoV capsid protein showed a highly conserved secondary structural elements, but highly active in VP1-U, leading end of VP2 and insertions between the strands of the betaG-H. This cued that HBoV inclined to immune evasion and infectant adaptive faculty.
Amino Acid Sequence ; Base Sequence ; Bocavirus ; classification ; genetics ; Capsid Proteins ; chemistry ; genetics ; Cloning, Molecular ; Conserved Sequence ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction

Objective To observe the effects of Gandouling on reactive oxygen species (ROS) level, the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA and protein of neural stem cells of the mice cultured in high concentration copper. Methods The model of neural stem cells of the mice was cultured in vitro with high concentration copper. The experimental rats were randomly divided into blank control group, model group, and Gandouling low-, medium-, and high-dose groups. Each medication group was given relevant concentration of Gandouling serum for gavage. The MTT was adopted to test proliferation level on neural stem cells; flow cytometer was used to examine the change of ROS level in cells; qPCR was used to measure the expression of Nrf2 mRNA;Western blot was used to measure the change of the level of protein Nrf2 in cells. Results Compared with the blank control group, the proliferation rate of neural stem cells was significantly decreased, ROS levels were significantly increased, and Nrf2 gene and protein expression was significantly decreased (P＜0.01). Compared with the model group, neural stem cells proliferation rate was significantly increased, ROS levels were significantly reduced, and Nrf2 gene and protein expression was significantly increased (P＜0.05, P＜0.01). Conclusion Gandouling can promote the proliferation of neural stem cells in mice by reducing ROS content in high copper-loaded mice and up-regulating Nrf2 expression.

Objective To dynamically analyze the evolutionary process of cerebral edema absorption and the level of local iron in rats with intra-cerebral hematoma by high-field strength 7 Tesla MRI and explore the characteristics and mechanism of secondary injury after intra-cerebral hematoma.Methods Sixteen adult SD rats (about 150 g) were randomly divided into experimental group (n=10) and control group (n=6).Rat models in the experimental group were established by performing injection of 50 μL their own venous blood into their right caudate nucleus accurately. Rats in the control group were used normal saline,instead.After that,head MRI (T2 and T2-star scans) was performed 1,2,3,7 and 14 d after the injection; their imaging features were compared. Results Nine rats in the experimental group survived and 1 died after the operation; in the early days (within 3 d), the T2 weighing imaging showed that the time of relaxation surrounding the hematoma was longer than that in control group,suggesting that the zone of the edema surrounding the hematoma became more clearly.In the early days (within 3 d),T2-weighted imaging was clear,and the time of relaxation surrounding the hematoma increased rapidly,steadily improved 3 d after the operation and reached its peak level 7 dafter the operation; the damage area absorption decreased steadily but turned widening 3 d later and reached the peak 7 d later.T2-star value reached the peak rapidly 3 d after the operation,and then,moderated the downturn.The rats in the control group showed no obvious signal changes under MRI,except those with needle tract injury. Conclusion Secondary injury after intra-cerebral hemorrhage shows a rapidly injury progress in the short terrn at first,and then,has intensify again after a stable period; the local iron diffusion trend is synchronized to the secondary injury,suggesting that iron may play a key role in the mechanism of secondary brain edema.

OBJECTIVETo understand the genotypes of human metapneumovirus (hMPV) and the genetic character of hMPV attachment protein G sequence in Hunan, China.

METHODS232 nasopharyngeal aspirates (NPA) samples from hospitalized children with acute respiratory infections were collected from Hunan, China in 2005. HMPV was detected. The full length of G glycoprotein genes were amplified and sequenced. Bioinformatics soft-wares were employed to analyze the sequences.

RESULTS17/232 (7.3%) were showed hMPV positive. And co-infection rate with other viruses is 35%. The diagnoses of these hMPV positive cases are pneumonia, bronchiolitis and bronchopneumonia. Phylogenetic analysis for G genes from 13 hMPVs revealed the existence of four major subgroups: A1, A2, B1, B2 in Hunan, China in 2005. There are four types of sequence lengths of hMPV G glycoprotein, which are 711, 675, 660, 696nt. It is different in potential N-linked glycosylation sites and number of cysteine residues among these hMPVs of Hunan, China and Beijing, China. Also it is different from those in Japan and North America.

CONCLUSIONThe investigation of hMPV from Hunan, China in 2005 revealed the high speed of genetic variation and the marked character of geographic epidemic differences.

Amino Acid Sequence ; Child ; China ; epidemiology ; Genotype ; Glycoproteins ; classification ; genetics ; Humans ; Metapneumovirus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; RNA, Viral ; genetics ; Respiratory Syncytial Virus Infections ; epidemiology ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; classification ; genetics