Investigation of Pb-B Bonding in PbB2(BO)n- (n = 0-2): Transformation from Aromatic PbB2- to Pb[B2(BO)2]-/0 Complexes with BB Triple Bonds.

Boron has been found to be able to form multiple bonds with lead. To probe Pb-B bonding, here we report an investigation of three Pb-doped boron clusters, PbB2-, PbB3O-, and PbB4O2-, which are produced by a laser ablation cluster source and characterized by photoelectron spectroscopy and ab initio calculations. The most stable structures of PbB2-, PbB3O-, and PbB4O2- are found to follow the formula, [PbB2(BO)n]- (n = 0-2), with zero, one, and two boronyl ligands coordinated to a triangular and aromatic PbB2 core, respectively. The PbB2- cluster contains a BB double bond and two Pb-B single bonds. The coordination of BO is observed to weaken Pb-B bonding but strengthen the BB bond in [PbB2(BO)n]- (n = 1, 2). The anionic [PbB2(BO)2]- and its corresponding neutral closed-shell [PbB2(BO)2] contain a BB triple bond. A low-lying Y-shaped isomer is also observed for PbB4O2-, consisting of a central sp2 hybridized B atom bonded to two boronyl ligands and a PbB unit.

[Mechanism of Xuebijing Injection in treatment of sepsis-associated ARDS based on network pharmacology and in vitro experiment].

The aim of this study was to investigate the effect and molecular mechanism of Xuebijing Injection in the treatment of sepsis-associated acute respiratory distress syndrome(ARDS) based on network pharmacology and in vitro experiment. The active components of Xuebijing Injection were screened and the targets were predicted by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). The targets of sepsis-associated ARDS were searched against GeneCards, DisGeNet, OMIM, and TTD. Weishengxin platform was used to map the targets of the main active components in Xuebijing Injection and the targets of sepsis-associated ARDS, and Venn diagram was established to identify the common targets. Cytoscape 3.9.1 was used to build the "drug-active components-common targets-disease" network. The common targets were imported into STRING for the building of the protein-protein interaction(PPI) network, which was then imported into Cytoscape 3.9.1 for visualization. DAVID 6.8 was used for Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment of the common targets, and then Weishe-ngxin platform was used for visualization of the enrichment results. The top 20 KEGG signaling pathways were selected and imported into Cytoscape 3.9.1 to establish the KEGG network. Finally, molecular docking and in vitro cell experiment were performed to verify the prediction results. A total of 115 active components and 217 targets of Xuebijing Injection and 360 targets of sepsis-associated ARDS were obtained, among which 63 common targets were shared by Xuebijing Injection and the disease. The core targets included interleukin-1 beta(IL-1ß), IL-6, albumin(ALB), serine/threonine-protein kinase(AKT1), and vascular endothelial growth factor A(VEGFA). A total of 453 GO terms were annotated, including 361 terms of biological processes(BP), 33 terms of cellular components(CC), and 59 terms of molecular functions(MF). The terms mainly involved cellular response to lipopolysaccharide, negative regulation of apoptotic process, lipopolysaccharide-mediated signaling pathway, positive regulation of transcription from RNA polyme-rase Ⅱ promoter, response to hypoxia, and inflammatory response. The KEGG enrichment revealed 85 pathways. After diseases and generalized pathways were eliminated, hypoxia-inducible factor-1(HIF-1), tumor necrosis factor(TNF), nuclear factor-kappa B(NF-κB), Toll-like receptor, and NOD-like receptor signaling pathways were screened out. Molecular docking showed that the main active components of Xuebijing Injection had good binding activity with the core targets. The in vitro experiment confirmed that Xuebijing Injection suppressed the HIF-1, TNF, NF-κB, Toll-like receptor, and NOD-like receptor signaling pathways, inhibited cell apoptosis and reactive oxygen species generation, and down-regulated the expression of TNF-α, IL-1ß, and IL-6 in cells. In conclusion, Xuebijing Injection can regulate apoptosis and response to inflammation and oxidative stress by acting on HIF-1, TNF, NF-κB, Toll-like receptor, and NOD-like receptor signaling pathways to treat sepsis-associated ARDS.

Clinical features of Epstein-Barr virus in the intestinal mucosa and blood of patients with inflammatory bowel disease.

BACKGROUND: The role of Epstein-Barr virus (EBV) in inflammatory bowel disease (IBD) remains to be elucidated. The aim of this study was to investigate the presence of EBV in the blood and intestinal mucosa of patients with IBD and evaluate the association between EBV positivity and IBD. METHODS: Patients with IBD, hospitalized between January 2015 and April 2018, were enrolled. The EBV-DNA load in blood samples from each subject was analyzed by quantitative real-time polymerase chain reaction. EBV-encoded small-RNA 1 (EBER-1) was detected by in-situ hybridization in intestinal mucosa tissue sections of patients with IBD. RESULT: EBV-DNA was detected in 48 out of 568 patients with IBD (8.4%), and EBER-1 positivity was detected in 27 of these patients (56.3%). Refractory IBD and severe mucosal inflammation were more common in patients with detectable levels of EBER-1 than in those without; the number of EBER-1-positive cells positively correlated with mucosal inflammation (P value < 0.05). Age (≥60 years old) and use of azathioprine were risk factors for EBV infection. There was no significant difference in clinical remission rate and surgical rate between the EBER-1 positive group and EBER-1 negative group, antiviral group and the non-antiviral group, among IBD patients who tested positive for EBV-DNA. CONCLUSION: Elderly patients with IBD, treated with azathioprine, are more susceptible to EBV positivity. Further, EBV mucosal detection correlated with the severity of mucosal damage and refractoriness, but not prognosis.

LncRNA DANCR-miR-758-3p-PAX6 Molecular Network Regulates Apoptosis and Autophagy of Breast Cancer Cells.

OBJECTIVE: This study set out to probe into the effects of long non-coding RNA (LncRNA) differentiation antagonizing non-protein coding RNA (DANCR) on apoptosis and autophagy of breast cancer (BC) cells. METHODS: The expression levels of DANCR, miR-758-3p and paired box 6 (PAX6) in BC tissues and cell lines were detected. The transcription and protein levels of PAX6, apoptosis-related factors (caspase-3, caspase-9, Bax/Bcl-2), and autophagy-related factors (LC3B, Atg5, Beclin-1) in BC cells were detected. The cell proliferation, apoptosis, autophagy and the regulatory relationship between genes and target genes were analyzed. RESULTS: DANCR and PAX6 were up-regulated in BC tissues and cell lines, while miR-758-3p was opposite. Down-regulating DANCR inhibited the malignant proliferation of BC cells and also promoted apoptosis and autophagy, which showed that caspase-3, caspase-9, Bax/Bcl-2, LC3B, Atg5 transcription and protein levels increased, while Beclin-1 transcription and protein levels decreased. DANCR regulated miR-758-3p in a targeted manner, and its over-expression could weaken the anti-cancer effect of miR-758-3p on BC cells. In addition, miR-758-3p also directly targeted PAX6, and knocking down its expression could weaken the inhibitory effect of down-regulating PAK6 on BC cell apoptosis and autophagy. We also found that DANCR acted as a competitive endogenous RNA sponge miR-758-3p, thus regulating the PAX6 expression. CONCLUSION: DANCR-miR-758-3p-PAX6 molecular network plays a key regulatory role in BC cell apoptosis and autophagy, which may provide reference for treating patients.

Clinical performance of automated chemiluminescent methods for anticardiolipin and anti-ß2-glycoprotein I antibodies detection in a large cohort of Chinese patients with antiphospholipid syndrome.

INTRODUCTION: To assess the clinical performance and correlations of automated chemiluminescence assay (CIA) and enzyme-linked immunosorbent assay (ELISA) for detecting antiphospholipid (aPL) antibodies in the diagnosis of antiphospholipid syndrome (APS). METHODS: The study recruited 505 subjects, including 192 with APS, 193 with connective tissue diseases other than APS, and 120 healthy donors. We measured anticardiolipin (aCL) and anti-ß2-glycoprotein I (anti-ß2GPI) antibodies IgG, IgM, and IgA in all the samples using both CIA and ELISA. RESULTS: Total agreement between the two methods ranged from 83.50% for anti-ß2GPI IgG antibodies to 92.76% for anti-ß2GPI IgM antibodies in all the groups. Anti-ß2GPI and aCL IgG assays showed the highest Spearman's rho coefficients (anti-ß2GPI IgG = 0.742, aCL IgG = 0.715). Anti-ß2GPI IgG CIA showed the highest sensitivity for diagnosis of APS at 80.21%, which was significantly higher than the sensitivity of anti-ß2GPI IgG ELISA (52.08%). For diagnosis of APS, anti-ß2GPI IgG CIA had the best discrimination power with the area under the curves (AUC) of 0.922, followed by aCL IgG CIA (AUC of 0.905). While the CIA AUC was slightly higher in all cases, the difference was not statistically significant. CONCLUSION: CIA measurements had a good agreement and correlation with comparative ELISA assays. The CIA anti-ß2GPI IgG however was significantly more sensitive for APS diagnosis. The two assay methodologies showed comparable predictive powers and support the value of the CIA method for improved diagnosis and management of patients with APS.

Causes of death in hospitalized patients with systemic lupus erythematosus: a 10-year multicenter nationwide Chinese cohort.

To estimate the mortality and describe the causes of death in a large multicenter cohort of hospitalized patients with SLE in China. This was a retrospective study of a nationwide SLE cohort (10 centers, 29,510 hospitalized patients) from 2005 to 2014 in China. Standardized mortality ratios (SMRs) were calculated for all death and were stratified by sex and age. Chi-square test was used to determine whether the major causes of death vary in age, sex, duration of SLE, disease activity, or medications. Comparison between dead patients and survival controls was used to identify the risk factors for mortality. Logistic regression analysis was used to evaluate the risk factors for mortality. A total of 360 patients died during the study period, accounting for 1.22%. The overall SMR was 2.13 (95% CI 1.96, 2.30), with a particularly high SMR seen in subgroups characterized by younger age. Infection (65.8%) was the most common cause of death, followed by lupus nephritis (48.6%), hematological abnormality (18.1%), neuropsychiatric lupus/NPSLE (15.8%), and interstitial pneumonia (13.1%). Cardiovascular disease and malignancy contributed little to the causes of death. Infection, in particular severe pulmonary infection, emerged as the foremost risk factor for mortality, followed by lupus encephalopathy. However, lupus nephritis and hematological abnormalities occurred more frequently in survival patients. SLE patients at a younger age of diagnosis have a poorer prognosis. Infection dominated the causes of death in recent China. Ethnicity and medications might account for the differences in causes of death compared with western populations.

Antiphosphatidylserine/prothrombin antibodies (aPS/PT) as potential diagnostic markers and risk predictors of venous thrombosis and obstetric complications in antiphospholipid syndrome.

BACKGROUND: The aim of the study was to determine the prevalence and clinical associations of antiphosphatidylserine/prothrombin antibodies (aPS/PT) with thrombosis and pregnancy loss in Chinese patients with antiphospholipid syndrome (APS) and seronegative APS (SNAPS). METHODS: One hundred and eighty six Chinese patients with APS (67 primary, 119 secondary), 48 with SNAPS, 176 disease controls (79 systemic lupus erythematosus [SLE], 29 Sjogren's syndrome [SS], 30 ankylosing spondylitis [AS], 38 rheumatoid arthritis [RA]) and 90 healthy donors were examined. IgG and IgM aPS/PT, IgG/IgM/IgA anticardiolipin (aCL) and IgG/IgM/IgA anti-ß2-glycoprotein I (anti-ß2GPI) antibodies were tested by ELISA. RESULTS: One hundred and sixty (86.0%) of APS patients were positive for at least one aPS/PT isotype. One hundred and thirty five (72.6%) were positive for IgG aPS/PT, 124/186 (66.7%) positive for IgM aPS/PT and 99 (53.2%) positive for both. Approximately half of the SNAPS patients were positive for IgG and/or IgM aPS/PT. Highly significant associations between IgG aPS/PT and venous thrombotic events (odds ratio [OR]=6.72) and IgG/IgM aPS/PT and pregnancy loss (OR=9.44) were found. Levels of IgM aPS/PT were significantly different in APS patients with thrombotic manifestations and those with fetal loss (p=0.014). The association between IgG/IgM aPS/PT and lupus anticoagulant (LAC) was highly significant (p<0.001). When both were positive, the OR for APS was 101.6. Notably, 91.95% (80/87) of LAC-positive specimens were positive for IgG and/or IgM aPS/PT, suggesting aPS/PT is an effective option when LAC testing is not available. CONCLUSIONS: Anti-PS/PT antibody assays demonstrated high diagnostic performance for Chinese patients with APS, detected some APS patients negative for criteria markers and may serve as potential risk predictors for venous thrombosis and obstetric complications.

IgMk paraprotein from gammopathy patient can bind to cardiolipin and interfere with coagulation assay: a case report.

BACKGROUND: The monoclonal gammopathies are a group of plasma-cell proliferative disorders characterized by the secretion of monoclonal immunoglobulin (M protein or paraprotein). Some rare cases have revealed the specific affinity of paraprotein as autoantibody. Here we report a patient with monoclonal gammopathy of undetermined significance (MGUS) accompanied by a remarkable increase of anticardiolipin antibody (aCL) and an extensively decreased coagulation factor activity, however, without any clinical signs of antiphospholipid syndrome (APS) and bleeding. RESULTS: Our further investigation indicated that IgMκ paraprotein of this patient possessed an antibody activity against phospholipids so as to bind to cardiolipin and interfere with coagulation assay in vitro. CONCLUSIONS: This case might be indicative that an abnormality of coagulation tests, disturbed by IgMκ paraprotein, does not predict a risk of bleeding in this patient.

Clinical characteristics and laboratory findings of 252 Chinese patients with anti-phospholipid syndrome: comparison with Euro-Phospholipid cohort.

This study aims to characterize the Chinese Han patients with anti-phospholipid syndrome (APS) and compare the data with those of the Euro-Phospholipid cohort. We conducted a single center study consisting of 252 patients with definite APS from 2000 to 2015. We analyzed the clinical and laboratory characteristics of our cohort and compared the data with those of the Euro-Phospholipid cohort. Our cohort consisted of 216 females and 36 males, with a mean age at entry into this study of 41 years (range 11-74 years). Of these patients, 69 (27.4%) patients had primary APS, and 183 (72.6%) had secondary APS (SAPS), including 163 (64.7%) patients had systemic lupus erythematosus (SLE). Thrombotic events occurred in 190 (75.4%) patients, and the most common ones were deep vein thrombosis (40.1%) and stroke (23.8%), which were similar to the reports of the Euro-Phospholipid cohort. In contrast, our cohort had less pulmonary embolism (6.7%). Among 93 females with 299 pregnancy episodes, the rates of early (<10 weeks) and late fetal loss (≥10 weeks) were, respectively, 37.8% and 24.4%. The latter was significantly higher than that of the Euro-Phospholipid cohort. Moreover, 7 APS nephropathy patients (characterized histopathologically by thrombotic microangiopathy) and 8 catastrophic APS patients were found in our cohort. Anti-cardiolipin antibodies (aCL) were detected in 169 (67.1%) patients, lupus anti-coagulant (LA) was detected in 83 (32.9%), and anti-ß2 glycoprotein I antibodies (anti-ß2GPI) in 148 (58.7%) patients. These results show that some clinical manifestations of APS may vary among different racial groups.

Expression changes and regulation of AR and IGF-1 in PC3 prostate cancer cells treated with sexual hormones and flutamide.

The study aims to investigate the changes and regulation of androgen receptor and insulin-like growth factor-1 in the PC3 prostate cells treated with 5α-dihydrotestosterone, estrone, and flutamide. The PC3 cells were cultured and treated with 5α-dihydrotestosterone, estrone, and flutamide. Immunocytochemistry and Western blot were used to detect the expression of androgen receptor and insulin-like growth factor-1. The androgen receptor expression was analyzed by Western blot and optic density scan in the presence or absence of various kinase inhibitors. The statistical calculations were performed with the statistics-analyzing software package SPSS 13.0. A P <0.05 was considered statistically significant. The concentrations of 5α-dihydrotestosterone and flutamide could almost not change the expression of androgen receptor and insulin-like growth factor-1 in PC3. But, the concentrations of estrone could increase the expression of androgen receptor and insulin-like growth factor-1 when PC3 cells are exposed to the studied concentration at various times. The expression of androgen receptor was regulated by the inhibitor of signal pathways of PI3, MEK1/2, and JUK. The expressions of androgen receptor and insulin-like growth factor-1 were influenced by estrone and were not influenced by 5α-dihydrotestosterone and flutamide in PC3 cells. And, the expression of androgen receptor was regulated by multiple signal pathways.

Polymorphism analysis and evaluation of nine non-CODIS STR loci in the Han population of Southern China.

BACKGROUND: Knowledge of allele and genotype frequencies is an essential prerequisite to the use of any human polymorphism in forensic work. AIM: To study the genetic polymorphism and evaluate the application value of nine STR loci. SUBJECTS AND METHODS: Genotyping of nine STR loci, including D11S2368, D12S391, D13S325, D18S1364, D22-GATA198B05, D6S1043, D2S1772, D7S3048 and D8S1132, of 1050 unrelated individuals was performed with the STR_Typer_10_v1 kit and Genetic Analyzer 3100 and analyzed with PowerState V12.xls and Arlequin ver 3.11 analyzing software. RESULTS: Allele frequency distribution was statistically analyzed and Hardy-Weinberg equilibrium determined. Several common parameters used in forensic sciences were found: the heterozygosity (H) ranged from 0.827 to 0.892; the matching probability (MP) ranged from 0.029 to 0.074; the power of discrimination (PD) ranged from 0.926 to 0.971; the power of exclusion (PE) ranged from 0.649 to 0.779; the polymorphic information content (PIC) ranged from 0.77 to 0.86; and the typical paternity index (TPI) ranged from 2.88 to 4.62. CONCLUSION: The results indicate that nine STR loci are high polymorphic among the Han population in Southern China. This set of polymorphic STR loci is a useful tool in forensic paternity testing and anthropological study.

Changes of androgen receptor and insulin-like growth factor-1 in LNCaP prostate cancer cells treated with sex hormones and flutamide.

Changes of androgen receptor (AR) and insulin-like growth factor-1 (IGF-1) were investigated in LNCaP cells treated with 5α-dihydrotestosterone (DHT), estrone and flutamide. Real-time PCR, immunocytochemistry and western blotting were used to detect the expression of AR and IGF-1 in the presence or absence of various kinase inhibitors. Low concentrations of DHT, estrone and flutamide increased the expression of AR and IGF-1, especially estrone, with concentration and time dependence. With DHT and flutamide, there was a significant alteration in AR expression (p<0.001). The results indicated expression of AR and IGF-1 genes to be influenced by DHT, estrone and flutamide in LNCaP cells, regulated by multiple signal pathways.

[The role of cell cycle arrest in radiosensitization of nasopharyngeal carcinoma cell line CNE1 by inhibiting ATM expression].

BACKGROUND & OBJECTIVE: Cell cycle regulation is one of the most important determinants to ionizing radiosensitivity of cells. ATM gene is closely related with DNA damage repair and cell cycle checkpoints control. We previously reported that suppressing ATM expression with antisense ATM RNA could enhance radiosensitivity of nasopharyngeal carcinoma (NPC) cell line CNE1. This study was to explore the involved changes of cell cycle and the mechanisms of cell cycle arrest. METHODS: ATM gene was constructed into retrovirus vector pDOR to form recombinant pDOR-atm. CNE1 cells were transfected with pDOR-atm (CNE1/pDOR-atm cells) or pDOR (CNE1/pDOR cells) and irradiated with X-ray. Cell cycle and cell apoptosis were detected by flow cytometry at different time points after irradiation. RESULTS: S phase arrest was detected at 1, 4, and 8 h after irradiation in both groups, and G2 arrest at 24, and 48 h, while no comparable G1 arrest and apoptosis were revealed. The mean percentage of S phase cells was lower, and G2 phase cells was higher in CNE1/pDOR-atm group than in CNE1/pDOR group (P<0.05). CONCLUSION: The mechanisms of cell cycle regulation in radiosensitized CNE1 cells by inhibiting ATM expression might be related with the decreased accumulation of S phase cells and increased accumulation of G2/M phase cells, while have no relationship with G1 arrest and apoptosis.

Development of a five ChX STRs loci typing system.

We investigated the polymorphism of five X-chromosomal short tandem repeat markers (ChrX STRs) loci (DXS7132, DXS981, DXS6803, DXS6809, and DXS6789) and their value for forensic applications. A fluorescent multiplex polymerase chain reaction (PCR) for amplifying five ChrX STRs loci in the same PCR reaction was set up. A total of 827 unrelated individuals of the Han nationality in China were tested. The results show that the five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing, particularly for difficult paternity-deficient cases.

[Development and forensic application of a pentaplex X-STR loci typing system].

A fluorescent multiplex PCR that simultaneously amplifies five X-chromosomal short tandem repeat (X-STR) loci (DXS6803, DXS981, DXS6809, DXS6789 and DXS7132)was set up to study their polymorphic nature and to determine its use in kinship tests for forensic cases. PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer with GeneMapper ID 3.1 Analysis Software. The pentaplex system gave satisfactory results as its sensitivity, reproducibility and unambiguous genotyping. About 20 ng DNA was routinely used, although 0.25 ng DNA was sufficient for allele typing. The results demonstrate that the multiplex system of the five X-STR loci provides a fast technology for forensic identification and paternity testing. The X-STR pentaplex system can complement the analysis of AS-STR and Y-STR efficiently, especially in complex cases of kinship testing.

[Polymorphism of 7 Y-STR loci in Chinese populations by multiplex PCR genotyping using fluorescein-labeled primers].

We reported the multiplex-PCR-based genotyping method for 7 Y-STR loci, including DYS456, DYS464a/b/c/d, DYS527a/b labeled with FAM (blue) and DYS531, DYS709, DYS448, DYS522 labeled with JOE (green). We investigated the haplotype distribution of these 7 Y-STR loci among 151 unrelated Han males in the Guangdong Province and 106 unrelated males in the Henan Province, and evaluated this method for forensic practice. The results showed that this method could successfully determine the genotypes using as little as 0.02 ng genomic DNA, and the male's Y-STR genotypes could be detected in a DNA mixture in which the ratio of male/female components was 1:150 (160 ng in total amount of DNA template). There were 150 and 105 haplotypes found of these 7 Y-STR loci in these two Chinese populations, out of them 149 and 104 haplotypes appeared only once, respectively. The haplotype diversity in the two populations were 0.999912 and 0.999820, respectively. The distribution variation of the 7 Y-STR haplotypes between Guangdong and Henan Chinese populations was statistically significant (P<0.001). Thus, the fluorescein-labeled multiplex-PCR genotyping of 7 Y-STR loci is a valuable tool for forensic medicine practice and for human anthropology study.

[Sequence analysis of DYS522 and DYS527 loci and their genetic polymorphism among Guangdong Han population].

Analyzed the sequence characteristics and the genetic polymorphism of two new Y-STR loci: DYS522 and DYS527, in 151 unrelated Han males in the Guangdong Province. The results show that the DYS522 locus consists of repeats of a core sequence (GATA), with the number of repeats ranging between 9 and 13. The DYS527 locus contains two copies of a sequence motif. This motif has the following modular structure: (GGAA)3...(GGAA)2...(GGAA)2...(GGAA)3...(GGAA)4...(GGAA)3...(GAAA)m(GGAA)n, where the value of (m + n) ranges between 18 and 26 among different individuals. A rare copy with 15.3 (m+n) repeats was found. Altogether, 63 different haplotypes of these two loci were identified. Of these, 29 occurred only once, with a frequency of 0.0066, and the most common haplotype occurred at a frequency of 0.0728. This system has a haplotype diversity (HD) of 0.9780, and a discriminating power (DP) of 0.9715. The analysis of 38 father/son pairs has detected no mutation event at the DYS522 and DYS527 loci within any pair. It is found that these two loci are specific to the human species. These results indicate the DYS522/DYS527 loci to be highly polymorphic and useful genetic markers in forensic science and human evolution studies.

[How to draw a conclusion in motherless parentage testing using short tandem repeats as genetic makers].

OBJECTIVE: To calculate the exclusion power of STR loci in motherless parentage testing and to discuss how to draw a conclusion if there are inconsistent loci. METHODS: Based on the law of inheritance and allele frequency, the powers of exclusion of STR loci in motherless parentage testing (PE(M)) were calculated. Based on the mean PE(M) and mutation rate of 13 CODIS loci. The probabilities of inconsistence under paternity and non-paternity were calculated respectively according to binomial theorem. RESULTS: The PE(M) of locus having co-dominate alleles could be calculated as: PE(M) = (i = 1)sigma (n) p i 2(1-p (i))2+ (i < j)sigma (n) 2p (i)p (j)(1-p (i)-p (j))2. According to the formula, the average PE(M) of 13 CODIS was 0.411. Based on the mean PE(M) and mutation rate, the likelihood ratio of true father to random man (paternity index) was got using binomial theorem. CONCLUSION: The conclusion in motherless parentage testing could be drawn based on the likelihood ratio (paternity index) derived from mean PE(M) and mutation ratio.

[Radiosensitization of nasopharyngeal carcinoma cell line CNE1 by inhibiting the expression of ATM/PI3K functional region].

BACKGROUND & OBJECTIVE: It is reported that ATM gene is closely correlated to cellular radiosensitivity in several malignant tumors. Suppression of ATM protein expression leads to cellular radiosensitization. This study was to determine whether this effect also exists in nasopharyngeal carcinoma (NPC) cell line CNE1 by inhibiting the expression of ATM protein through antisense RNA of ATM/PI3K region, which is the most important functional fragment of ATM gene. METHODS: The recombinant pDOR-atm expressing antisense RNA of ATM/PI3K segment was constructed with retroviral vector pDOR. CNE1 cells were stably transfected with pDOR-atm by cationic liposome and named CNE1/pDOR-atm. Semi-quantitive RT-PCR was used to detect the level of ATM mRNA. Flow cytometry (FCM) was employed to analyze the percentage of positive cells and mean fluorescence density of protein expressing ATM. Cellular radiosensitivity was evaluated by colony survival assay (CSA) and linear-quadratic model in both CNE1/ pDOR-atm and control cells. RESULTS: The level of ATM mRNA index (RI) was 0.23+/-0.02 in CNE1/pDOR-atm group, and 0.51+/-0.03 in control group (P<0.05). The percentage of positive cells and mean fluorescence density of proteins expressing ATM were 70.8% and 1.81+/-0.12 in CNE1/pDOR-atm group, while 99.3% and 4.51+/-0.18 in control group(P<0.01). The expression of ATM mRNA and protein was inhibited by antisense RNA. The alpha value (one function from linear-quadratic model) of CNE1/pDOR-atm group was 0.40 Gy(-1), while it was 0.36 Gy(-1) in control group. The radiosensitizing ratio of surviving fraction at 2 Gy (SF(2)) was 3.0, indicating that antisense RNA group was more radiosensitive to X-ray than the controls. CONCLUSION: NPC cell line CNE1 could be radiosensitized by the down-regulation of ATM/PI3K expression.

[Genetics of heteroplasmy in the mtDNA control region among the Chinese Han population].

OBJECTIVE: To explore the distribution and genetic pattern of heteroplasmy of mtDNA control region among Chinese Han population. METHODS: The human mtDNA control region was amplified into 6 amplicons overlapped partially each other. Then these amplicons were analyzed by DHPLC which we developed to detect low heteroplasmic signals. RESULTS: There were 51 heteroplasmic cases (34%) found from different tissues of 150 unrelated individuals of the Chinese Han population. mtDNA heteroplasmy shows non-uniform distribution in various tissues. The highest occurrence of heteroplasmy was in brain tissues (50/150) and myocardium (48/150), the lowest was in bone tissues (22/150). 36 sites of heteroplasmy were identified in our samples. Three sites of mtDNA heteroplasmy rarely co-existed in one individual. No sex differences were detected in the frequency of mtDNA heteroplasmy. No change in the mtDNA heteroplasmy profile was detected of blood samples from the same individuals within 2 years. Individuals older than 41 years showed a heteroplasmy frequency significantly higher than their younger counterparts. Members from the same maternal pedigree in a family can share the same sites of mtDNA heteroplasmy but may have different heteroplasmy contents at those sites. CONCLUSION: DHPLC is a highly sensitive technique in detecting heteroplasmy. mtDNA heteroplasmy widely exists in the Chinese Han population. The results shown here could potentially have a guidable value in forensic individual identification and parentage testing.