[Effects of slope direction on soil nutrient and its ecological stoichiometry in bamboo forest]. / å¡å‘å¯¹æ¯›ç«¹æž—åœŸå£¤å »åˆ†åŠå ¶ç”Ÿæ€åŒ–å­¦è®¡é‡ç‰¹å¾çš„å½±å“.

We analyzed the effects of slope direction on soil nutrients and ecological stoichiometry by collecting soil samples from different slope directions (shady slope and sunny slope) of the bamboo forest in Longyou County, Zhejiang Province. The results showed that soil nutrients were affected by slope direction and soil depth. The nutrients level of soils in the sampling area showed the trends of shady slope > sunny slope, and surface soil > bottom soil. Compared to sunny slope, the cation exchange capacity (CEC), the contents of total organic carbon, total nitrogen, alkaline hydrolyzed nitrogen, available phosphorus, total potassium and available potassium of shady soils significantly increased by 43.7%, 103.8%, 92.0%, 75.5%, 22.4%, 89.4% and 240.7%, respectively. There was no significant difference in total phosphorus contents between shady slope and sunny slope. At all soil layers, there was no significant difference of C/N ratio between shady and sunny slopes. The average C/P ratio of shady slope was 180.8%, 42.0% and 54.3% higher than that of sunny slope at 0-20 cm, 20-40 cm and 40-60 cm, respectively. At each soil layer, the average C/K and N/K ratios between shady and sunny slopes had no significant difference. The average C/K and N/K ratios of shady slope and sunny slope were all significantly different among the three soil layers. In the shady slope, the contents of soil organic carbon showed significantly positive correlation with total nitrogen, total phosphorus, total potassium, and soil available nutrients. Overall, soil nutrients and ecological stoichiometry characteristics of shady slope of bamboo forest were superior to those of sunny slope.

Association of IL-10 (-819T/C, -592A/C and -1082A/G) and IL-6 -174G/C gene polymorphism and the risk of pneumonia-induced sepsis.

CONTEXT: Association between inherited variants and the risks of sepsis is controversial. OBJECTIVE: To evaluate the risk of pneumonia-induced sepsis by examining its linkage with polymorphisms of IL-6 and IL-10. MATERIALS AND METHODS: Samples were obtained from 188 pneumonia-induced sepsis patients, 162 pneumonia patients and 200 healthy controls. RESULTS: Subjects with IL-10 -1082 AA genotypes and IL-6 -174 CC genotype had a higher risk of sepsis and increased mRNA levels. CONCLUSION: The variants of IL-10 -1082 A allele and IL-6 -174 C allele contributed to an increased risk of pneumonia-induced sepsis.

Premalignant alteration assessment in liver-like tissue derived from embryonic stem cells by aristolochic acid I exposure.

The in vitro predictive evaluation of chemical carcinogenicity based on hepatic premalignance has so far not been established. Here, we report a novel approach to investigate the premalignant events triggered by human carcinogen aristolochic acid I (AAI) in the liver-like tissue derived from mouse embryonic stem cells. By AAI exposure, the liver-like tissue exhibited the paracrine interleukin-6 phenotypic characteristics. Hepatocytes expressed STAT3/p-STAT3, c-Myc and Lin28B in parallel. Some of them displayed the dedifferentiation characteristics, such as full of α-fetoprotein granules, increase in size, and nucleocytoplasmic shuttle of Oct4. When these cells were injected into mice, the xenografts mostly displayed the uniform area of hepatic-like tissue with malignant nuclei. The hepatic malignant markers, α-fetoprotein, cytokeratin 7 and cytokeratin 19, were co-expressed in albumin-positive areas, respectively. In conclusion, we established an approach to predict the hepatic premalignance triggered by carcinogen AAI. This premalignant assay system might aid to evaluate the effects of potential carcinogens in liver, and probably to screen the protecting against hepatocarcinogenic efficacy of pharmaceuticals in vitro.

Association of tumor necrosis factor α -308G/A and interleukin-6 -174G/C gene polymorphism with pneumonia-induced sepsis.

PURPOSE: Sepsis is a lethal outcome of the inflammation and coagulation process. Human interleukin (IL)-6 and tumor necrosis factor (TNF) α are well-known inflammation factors closely associated with sepsis. In the present study, we aim to investigate the association of promoter-region polymorphisms IL-6 (-174G/C) rs1800795 and TNF-α (-308G/A) rs1800629 with pneumonia-induced sepsis. MATERIALS AND METHODS: A total of 277 Chinese patients with severe pneumonia-induced sepsis were recruited into this study. All study participants were admitted to the intensive care unit until discharge or death in the First Affiliated Hospital of Zhengzhou University from July 2010 to July 2014. The patients were classified as severely septic, septic shock, and mortality. Clinical data and demographic information were recorded. TaqMan genotyping was performed to detect single nucleotide polymorphism distribution. RESULTS: The genotype results demonstrated that carriers of the TNF-α rs1800629 A allele had a 4.28-fold higher risk for septic shock (adjusted odds ratio [OR], 4.28; 95% confidence interval [CI], 2.24-8.18; P < .01) compared with severe sepsis, and carriers of the IL-6 rs1800795 C allele had a 2.42-fold higher risk for septic shock (OR, 2.42; 95% CI, 1.08-5.45; P < .01) compared with severe sepsis. No significant difference of SNP distribution was found between the survivors and the nonsurvivors. After the results were adjusted for age and the outcomes of blood cultures, a multivariate logistic regression analysis showed similar results. Individuals with the TNF-α 308 rs1800629 A allele (adjusted OR, 2.96; 95% CI, 1.30-7.87) or the IL-6 rs1800795 C allele (adjusted OR, 1.87; 95% CI, 1.03-3.61) had a higher prevalence of septic shock. However, these SNP distribution differences were not associated with mortality. CONCLUSIONS: In intensive care unit patients, the TNF-α -308A allele and the IL-6 rs1800795 allele variants were susceptibility risk factors for septic shock induced by pneumonia.

Epstein-Barr virus interactions with the Bcl-2 protein family and apoptosis in human tumor cells.

Epstein-Barr virus (EBV), a human gammaherpesvirus carried by more than 90% of the world's population, is associated with malignant tumors such as Burkitt's lymphoma (BL), Hodgkin lymphoma, post-transplant lymphoma, extra-nodal natural killer/T cell lymphoma, and nasopharyngeal and gastric carcinomas in immune-compromised patients. In the process of infection, EBV faces challenges: the host cell environment is harsh, and the survival and apoptosis of host cells are precisely regulated. Only when host cells receive sufficient survival signals may they immortalize. To establish efficiently a lytic or long-term latent infection, EBV must escape the host cell immunologic mechanism and resist host cell apoptosis by interfering with multiple signaling pathways. This review details the apoptotic pathway disrupted by EBV in EBV-infected cells and describes the interactions of EBV gene products with host cellular factors as well as the function of these factors, which decide the fate of the host cell. The relationships between other EBV-encoded genes and proteins of the B-cell leukemia/lymphoma (Bcl) family are unknown. Still, EBV seems to contribute to establishing its own latency and the formation of tumors by modifying events that impact cell survival and proliferation as well as the immune response of the infected host. We discuss potential therapeutic drugs to provide a foundation for further studies of tumor pathogenesis aimed at exploiting novel therapeutic strategies for EBV-associated diseases.

[Research progress on prognostic markers of chronic lymphocytic leukemia].

Chronic lymphocytic leukemia (CLL) remains the most common adult leukemia. The recent progress on research of molecular and cellular genetics of CLL promotes the development of the diagnosis, treatment and prognosis for CLL patients. IGVH gene mutation status is the most important prognostic marker for CLL patients. Zeta-chain-associated protein kinase (ZAP-70) can be used as a surrogate marker for IGVH mutation status. CD38 is a type II transmembrane glycoprotein promoting B cell activation and proliferation, which can improve the survival of CLL cells and enhance their proliferation, so it also can be used as an independent prognostic indicator for CLL. Chromosome aberrations are found in more than 80% of CLL cases. The most frequent abnormalities are losses of chromosomal material, with deletions in band 13q14 being the most common. The most common gains of chromosomal material are trisomies 12q. Human leukocyte antigen G (HLA-G) is a non-classical HLA-I gene. Increased expression of HLA-G leads to the malignant progression of CLL, significantly shortens survival, indicating HLA-G might serve as a prognostic marker in CLL. Toll-like receptors (TLA) are important component of natural immunity. The combination of TLR agonists and release chemotherapy, monoclonal antibodies and tumor vaccines would bring a breakthrough for the treatment of CLL.

[Clonality analysis and mutational status of IgVH gene in Hodgkin variant of Richter syndrome].

OBJECTIVE: To detect the clonal relationship, the rearrangement, and the mutational status of IgVH gene; the influence of these molecular characteristics on the clinical outcome in Hodgkin variant of Richter syndrome; and the possible molecular pathogenesis in this transformation. METHODS: The clonal rearrangements and mutational status of IgVH genes were analyzed in Hodgkin variant of Richter syndrome and B-CLL with Reed-Stemberg (R-S)-like cells by GeneScan analysis and sequencing. Semi-nest PCR based on laser capture microdissection was utilized to compare the clonal relationship between B-CLL and R-S/R-Slike cells. Immunohistochemical staining was used to detect the different expressions of ZAP70, p53, IRF-4 and LMP1 in the two components. RESULTS: (1) 5/6 B-CLL cases transformed to Hodgkin lymphoma (HL)/R-S-like cells carried the mutated IgVH genes; (2) 2 cases of R-S cells and 1 case of R-S-like cells were clonally distinct from B-CLL clone and express LMP1, whereas 1 case of R-S-like cells was relating to the surrounding B-CLL cells and did not express LMP1; (3) 2/6 B-CLL cases transformed to HL convey VH4-34 and VH3-48 respectively. CONCLUSIONS: (1) Richter transformation to HL/R-S-like cells evolves from the B-CLL which originates from the germinal center or post germinal center B cells, indicating that different lymphoma cells of different subtypes in Richter syndrome come from different B cell lineage and possibly involve a different pathogenesis and pathway; (2) HL and R-S-like cells evolve from either the B-CLL clone or may develop as a clonally unrelated lymphoma, the independent secondary malignancies are appear to be EBV-positive, possibly as a consequence of the underlying immunodeficiency; (3) The biased usage of IgVH genes suggested a role of antigens involved in the HL variant of Richter syndrome.

[Clonality analysis and mutation status of IgVH genes in classic Richter's syndrome].

OBJECTIVE: To study the clonal rearrangements and mutation status of IgVH genes in classic Richter's syndrome, the relationship between molecular findings of IgVH gene and clinical outcome, and to deciper the possible molecular mechanism of transformation. METHODS: The clonal rearrangements and mutation status of IgVH genes were analyzed in cases of classic Richter's syndrome by Genescan and sequencing. Immunohistochemical study for zeta-chain associated protein kinase 70 kDa (ZAP70), p53 and interferon regulation factor 4 (IRF-4) was also performed. RESULTS: Samples of 18 cases of B-chronic lymphocytic leukemia (B-CLL)/ diffuse large B-cell lymphoma (DLBCL,78. 3%) had identical tumor cell clones, whereas DLBCL developed as a clonally independent neoplasm in 5 patients (21.7%). Among the clonally related group, 12 cases carried unmutated VH genes in both B-CLL and DLBCL components and VH3-23, VH3-74 and VH1-2 were accounted for the B-CLL transformation to DLBCL. Immunohistochemical study showed that the transformed DLBCL expressed CD5 in 32.1% of cases, CD23 in 14.3%, ZAP70 in 23.8%, p53 in 80.6% and IRF-4 in 82.6% of the cases respectively. Follow-up data were available in 17 patients with classic Richter's syndrome. The median survival period was 7 months. No significant difference in survival rate was obtained between the clonally related or unrelated groups, between IgVH gene mutated or unmutated groups, and between the groups with or without expression of ZAP70, p53 and IRF-4. CONCLUSIONS: The ratio of clonally related transformed DLBCL from B-CLL to clonally unrelated DLBCL is 2:1. Clonal transformation to DLBCL predominantly occurs in B-CLL patients carrying unmutated IgVH genes. The biased IgVH gene usage suggests antigens are involved in classic Richter's syndrome. Molecular differences of IgVH genes and very poor clinical outcome of this group of transformed DLBCL indicate that there cases may be regarded as a distinct subset of DLBCL.

A novel microsatellite within intron 8 in caspase10 gene in Chinese of Han nationality.

OBJECTIVE: To investigate polymorphisms in the coding exons and their splicing areas of caspase10 gene (CASP10) in Chinese of Han nationality. METHODS: Polymerase chain reaction (PCR), denaturing high-performance liquid chromatography (DHPLC), direct sequencing and clonal sequencing were used to analyze the exon 9 and its flanking sequences. RESULTS: In 70 blood samples of Chinese subjects researched the known single nucleotide polymorphisms (SNPs) in the exon 9 of CASP10 gene published in dbSNP of NCBI were not identified. We found a short sequence with more than ten continuous and repeated T nucleotides within the intron 8 near the exon 9 and the length of repeated sequence nucleotides T was different in samples. Clonal sequencing analysis indicated that it was a microsatellite of single nucleotide-repeated sequence. The recurrence and specificity of same result was further confirmed by PCR with high fidelity polymerase and sequencing. Furthermore all of 70 blood samples detected by DHPLC showed heterozygous profiles. We named it as IVS8-13(T)n temperately. CONCLUSION: Our research suggested that the site be a microsatellite of single nucleotide-repeated sequence. Meanwhile, our data further showed that it was quite different from that by querying SNP database of non-Chinese population in NCBI, in the same region a SNP of type of insertion deletion existed. It could be inferred that the difference between the populations might be associated with the genetic characteristics of ethnics. The significance of the microsatellite in CASP10 gene in Chinese of Han nationality remains to be elucidated.

[BCL-2/IgH translocation in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area].

OBJECTIVE: To investigate the relationship between the frequency of BCL-2/IgH rearrangement in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area and the low incidence of follicular lymphoma (FL). METHODS: Nested-PCR and direct DNA sequencing were used to detect the Bcl-2/IgH rearrangement in peripheral blood cells of 196 healthy individuals. DNA sequences involved were then searched and aligned in NCBI database to confine the broken points in major breakpoint region and the IgH segments involved. RESULTS: First, in this sample the frequency of BCL-2/IgH translocation in Chinese individuals of Han nationality located in Zhejiang area is 9.66%, being much lower than that in North America and Europe countries. Second, the breakpoints tend to fall into 3 clusters: 3055, 3116 and 3165 bp. Usage of J6 segment is most common. Third, There are different subclones of BCL-2/IgH rearrangements in the same individual. CONCLUSION: The low frequency of BCL-2/IgH translocation in healthy Chinese individuals of Han nationality located in Zhejiang area may be one of the reasons for the difference in the incidence of FL between China and Western countries.

[Detection of IgH gene rearrangement in B-cell non-Hodgkin's lymphoma and application of gel-scan method].

OBJECTIVE: To develop a protocol for gene rearrangement study in non-Hodgkin's lymphoma (NHL) by PCR-directed gel-scan method and to set up quantitative criteria for IgH gene rearrangement which can be applied in the follow up of lymphoma patients. METHODS: IgH gene rearrangement studies were carried out in 96 cases of B-cell NHL. The detection rate of clonality was evaluated. Sixty-five cases of IgH gene rearranged cases proven by FR3A-directed PCR and PAGE and 8 cases of benign lymphoid tissues (5 cases of reactive lymphoid hyperplasia, 3 cases of chronic tonsillitis), 5 cases of normal peripheral blood mononuclear cells were analyzed by gel-scan method and the proportion of h1/h2 (heights of peak1 and peak2 of gel-scan) was calculated. RESULTS: The detection rate of IgH gene clonality was up to 68% using primer FR3A in the 96 B-cell NHL cases. The detection rate was up to 61% using primer FR2A. With a combination of primers FR3A and FR2A, the detection rate increased to 83%. Gel-scan curve showed that the value of h1/h2 was greater than 3 in all the 65 cases with IgH gene rearranged. In the 8 benign lymphoid tissue cases showed h1/h2 < 1.5, 5 cases with normal peripheral blood mononuclear cells showed a bell-shaped curve. CONCLUSIONS: In the gel-scan curve of gene rearrangement studies in non-Hodgkin's lymphoma samples, the value of h1/h2 greater than 3 represents a true clonal proliferation. The peaks with relative heights less than 1.5 may not be significant and likely represent polyclonal cell population. A value between 1.5 and 3 however requires clinical follow-up. The success rate of rearrangement studies in B-cell NHL can be increased by using a combination of primers FR3A and FR2A.

[Study of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphoma].

OBJECTIVE: To investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance. METHODS: PCR analysis of monoclonal, polyclonal and dual rearrangements of IgH and TCR gamma, TCR beta genes was carried out in 125 cases of NHL to evaluate the rate of dual rearrangements, immunohistochemistry was performed for a Ki67 protein expression in 117 cases and the proliferation index was calculated. The relationship between antigen receptor gene rearrangements and proliferation index was analyzed. RESULTS: Combination of the two pairs of IgH gene primers with the multiplex PCR for TCR gamma and TCR beta gene revealed dual rearrangements in 8% (8/96) of B-NHL, 17% (5/29) of T-NHL. In B cell NHL, IgH gene monoclonal, dural and polyclonal rearrangements were identified in 65, 8 and 15 cases respectively, while in T-cell NHL, they were in 15, 5 and 9 cases, respectively. There was no significant difference between proliferation index and monoclonal, dual, polyclonal rearrangements in both B-NHL and T-NHL by One-way test. CONCLUSION: Dual rearrangements in NHL are not rare and have no relationship with proliferation index.