Developmental validation of the DNATyper™Y26 PCR amplification kit: An enhanced Y-STR multiplex for familial searching.

The DNATyper™Y26 PCR Amplification kit, which including 26 low-medium mutating Y-STRs, is designed for Y-STR familial searching casework. The kit combines nine new Y-STR loci in addition to the 17 Y-STR loci from the commercially available AmpFlSTR®Yfiler® kit. The validation of the DNATyper™Y26 kit was performed in terms of technical index, including accuracy, stability, species specificity, sensitivity, adaptability for various samples, and mixture. Further, mutations of the 26 Y-STRs were analyzed by 1167 DNA-confirmed father-son pairs, and the results indicated that these loci had low or medium mutation rates. Furthermore, these Y-STRs loci were also tested in 1072 random male samples from Henan, Shanxi, Inner Mongolia, and Chongqing in China, showing their high power for forensic discrimination in the Chinese population. Thus, the DNATyper™Y26 PCR Amplification kit is a powerful tool for 'Y-STRs familial searching' in actual sexual-assault cases, indicating its unique advantage in familial searching due to Y-STR loci with only low-medium mutation rates.

[Succession of rotifer community and its relationship with environmental factors in a new Estuarial Landscape River, Shanghai].

The community structures of rotifers were surveyed in Lin'gang B Gang River, a new estuarial river in Shanghai, from December 2011 to November 2012. A total of 23 rotifer species were identified, of which 19 species were the indicator of environmental pollution. Rotifer species' number rose monthly from 2 to 13. A conspicuous succession of dominant species was observed from a single dominant species (Keratella cochlearis) to several species. The rotifer community structure of Lin'gang B Gang River differs with that of the outer water body (Lu Chaoyin River). Pearson analysis showed that the rotifers' density had a significant correlation with water temperature and chlorophyll-a (P <0. 05) , but had no significant correlation with salinity; the densities of dominant species (except Keratella cochlearis) presented negatively correlations with concentration of nutrients and salinity (P < 0. 05). The low species similarity index between every month demonstrated that aquatic ecosystem of Lin'gang B Gang River was in the unstable state. However, the increase of Margalef index and Shannon-Wiener index of rotifers and the decline of E/O value monthly showed the water environment of Lin'gang B Gang. River was improved.

Synthesis and biological activity of 4-substituted benzoxazolone derivatives as a new class of sEH inhibitors with high anti-inflammatory activity in vivo.

A series of novel 4-substituted benzoxazolone derivatives was synthesized, characterized and evaluated as human soluble epoxide hydrolase (sEH) inhibitors and anti-inflammatory agents. Some compounds showed moderate sEH inhibitory activities in vitro, and two novel compounds, 3g and 4j, exhibited the highest activities with IC50 values of 1.72 and 1.07 µM, respectively. Structure-activity relationships (SARs) revealed that introduction of a lipophilic amino acid resulted in an obvious increase in the sEH inhibitory activity, especially for derivatives containing a phenyl (3d, IC(50) = 2.67 µM), pyrrolidine (3g, IC(50) = 1.72 µM), or sulfhydryl group (3e, IC(50)=3.02 µM). Several compounds (3a-3g) were tested in vivo using a xylene-induced ear edema mouse model. Three compounds (3d, 3f, and 3g) showed strong anti-inflammatory activities in vivo which were higher than that of Chlorzoxazone, a reference drug widely used in the clinic. Our investigation provided a novel type of sEH inhibitor and anti-inflammatory agent that may lead to the discovery of a potential candidate for clinical use.

Apoptosis induced neurotoxicity of Di-n-butyl-di-(4-chlorobenzohydroxamato) Tin (IV) via mitochondria-mediated pathway in PC12 cells.

The severe toxicity of antitumor organotin (IV) compounds limits their application in clinic, however, the toxic mechanism is still unclear. Di-n-butyl-di-(4-chlorobenzohydroxamato) Tin (IV) (DBDCT), an antitumor agent with high activity and obvious neurotoxicity was chosen as a typical diorganotin (IV) compound to investigate its neurotoxic mechanism using PC12 cells and comprehensive methods. Treatment with DBDCT resulted in a dose- and time-dependent growth inhibition of PC12 cells. The changes in cell morphology were observed using light microscopy, fluorescence microscopy and transmission electron microscopy. PC12 cell apoptosis induced by DBDCT was confirmed by annexin V/propidium iodide staining, and characterized by cleavage of caspase-9 and caspase-3 proteins. DBDCT induced the release of cytochrome c from the mitochondria to the cytosol and the generation of reactive oxygen species. DBDCT up-regulated the expression of Bax, down-regulated the expression of Bcl-2, and significantly increased the ratio of Bax/Bcl-2. DBDCT also caused the phosphorylation of JNK and p38(MAPK). In rats exposed to DBDCT, apoptosis was also observed in brain, as shown by the detection of cleaved caspase-9 and caspase-3 proteins and increased TUNEL positive staining. In conclusion, the results demonstrated that DBDCT caused the neurotoxicity by inducing apoptosis via mitochondria-mediated pathway.

Identification and characterization of γ-aminobutyric acid uptake system GabPCg (NCgl0464) in Corynebacterium glutamicum.

Corynebacterium glutamicum is widely used for industrial production of various amino acids and vitamins, and there is growing interest in engineering this bacterium for more commercial bioproducts such as γ-aminobutyric acid (GABA). In this study, a C. glutamicum GABA-specific transporter (GabP(Cg)) encoded by ncgl0464 was identified and characterized. GabP(Cg) plays a major role in GABA uptake and is essential to C. glutamicum growing on GABA. GABA uptake by GabP(Cg) was weakly competed by l-Asn and l-Gln and stimulated by sodium ion (Na(+)). The K(m) and V(max) values were determined to be 41.1 ± 4.5 µM and 36.8 ± 2.6 nmol min(-1) (mg dry weight [DW])(-1), respectively, at pH 6.5 and 34.2 ± 1.1 µM and 67.3 ± 1.0 nmol min(-1) (mg DW)(-1), respectively, at pH 7.5. GabP(Cg) has 29% amino acid sequence identity to a previously and functionally identified aromatic amino acid transporter (TyrP) of Escherichia coli but low identities to the currently known GABA transporters (17% and 15% to E. coli GabP and Bacillus subtilis GabP, respectively). The mutant RES167 Δncgl0464/pGXKZ9 with the GabP(Cg) deletion showed 12.5% higher productivity of GABA than RES167/pGXKZ9. It is concluded that GabP(Cg) represents a new type of GABA transporter and is potentially important for engineering GABA-producing C. glutamicum strains.

[Aidi Injection as an adjunct therapy for non-small cell lung cancer: a systematic review].

OBJECTIVE: To assess methodological quality of clinical studies using Aidi Injection as an adjunct therapy for non-small cell lung cancer (NSCLC) and to evaluate the effects of Aidi Injection. METHODS: PubMed (1980-2008), Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 3, 2008), EMBASE (1984-2008), CancerLit (1996-2003), CBMdisc (1980-2008), CNKI database (1980-2008), Wanfang database (1980-2008), and Chongqing VIP database (1980-2008) were searched. The authors also hand-searched Chinese periodicals potentially related to the question. All randomized controlled trials comparing Aidi Injection with other treatment methods of NSCLC were included. Two reviewers selected studies, assessed the quality of studies, and extracted the data independently. The included studies were appraised and analyzed by Cochrane Collaboration Review Manager 5.0. RESULTS: Fourteen randomized controlled trials were included in the meta-analysis, but the quality of reports of the 14 included studies was poor. Aidi Injection combined with cobalt-60 or navelbine and platinol (NP) showed statistically significant differences in improving the response rate as compared with single use of cobalt-60 (P=0.000 2) or NP (P=0.04), and the relative risk (RR) and 95% confidence interval (CI) were 1.93 [1.36, 2.72] and 1.18 [1.00, 1.38], respectively. However, Aidi Injection combined with etoposide and platinol (EP), taxinol and platinol (TP) or gamma knife showed no significant differences as compared with single use of EP (P=0.60), TP (P=0.16) or gamma knife (P=0.34), respectively. The RR and 95% CI of EP, TP and gamma knife were 1.17 [0.65, 2.09], 1.27 [0.91, 1.78] and 1.08 [0.92, 1.26] respectively. Six studies indicated that Aidi Injection combined with NP or gamma knife could improve the quality of life.Six studies showed that Aidi Injection combined with NP or TP could improve the bone marrow hematopoietic function.Three studies indicated that Aidi Injection combined with NP could improve the immune function. Three studies showed that the combined therapy could not improve 1-, 2-, and 3-year survival rates. CONCLUSION: The results of meta-analysis indicate that Aidi Injection may have adjuvant therapeutic effects in treatment of NSCLC patients, but the sample size is too small and with poor quality, and the existence of publication bias is found.The effects of Aidi Injection need to be confirmed by large multicenter randomized controlled trials.