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Despite concerns about overfishing and the potential impact of release programs on wild populations, our study of 3116 individuals from 13 wild populations and 2787 individuals from two cultured populations in Zhejiang and Fujian provinces spanning 2008 to 2023 reveals a relatively stable genetic diversity in Larimichthys crocea . Surprisingly, the genetic diversity of wild large yellow croaker populations has remained consistent over the years, suggesting minimal influence from population declines due to overfishing. With the exception of populations in Sansha Bay and Luoyuan Bay, no significant genetic differences were observed among wild populations, indicating a single panmictic genetic population across the East and South China seas. Notably, significant genetic differentiation exists between cultured and wild populations, suggesting a possible limited genetic adaptation of cultured-released individuals to the wild environment. The genetic differences observed between the Sansha Bay, with its adjacent Luoyuan Bay populations, and other wild populations underscore the dual effects of habitat environment and farming activities on the genetic structure of large yellow croaker. Our findings suggest that, despite declines in population numbers due to overfishing and expands extensive cultured releases, the genetic diversity of L. crocea populations remains largely unaffected. Moreover, the L. crocea population along the Chinese coast appears to form a single panmictic population with considerable genetic diversity., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd.)

Hutuo River Basin straddles Shanxi and Hebei provinces, and Hutuo River was once cut off due to economic development and urban expansion after 2000; however, with the national emphasis on ecological civilization and the implementation of the South-North Water Diversion Project, the ecological protection of Hutuo River Basin has been significantly improved. MODIS data, Landsat data, and night light remote sensing data were selected based on the google earth engine (GEE) platform, and a new evaluation index system was generated by combining the biological richness index, vegetation cover index, land stress index, and pollution load index in the ecological environment index (EI) and the humidity index in the remote sensing ecological index (RSEI), using the variation coefficient method and entropy weighting method to assign weights to these indices. An ecological environment evaluation model was constructed to evaluate and classify the ecological environment quality of Hutuo River Basin from 2000 to 2020, and the driving factors were interpreted by using geographic probes. The results showed that:① on a time scale, the ecological environment of Hutuo River Basin was in a decline period from 2000 to 2015 and a recovery period from 2015 to 2020. From a grid scale, the ecological environment quality in the central part of the basin showed a state of improvement year by year, and in the western and eastern parts of the basin, the ecological environment quality in the decline period decreased year by year, whereas the ecological environment quality in the recovery period improved. ② Hot spot analysis showed that the spatial distribution of the ecological environment quality in Hutuo River Basin was high in the middle and low on both sides. Cold spot regions were mainly located in major cities and towns in the eastern and southern parts and scattered in the river valley area on the west side. ③ Geodetection analysis showed that the single factor detection drivers were mainly population density, vegetation net primary productivity (NPP), fractional vegetation cover (FVC), and geomorphological type. The dominant factor of cross-detection was "geomorphological type + FVC." With the deepening of ecological civilization construction and the implementation of Hutuo River Protection Regulations, in combination with different factors such as the natural environment and social characteristics in this basin, the research on ecological environment evaluation in Hutuo River Basin can provide data support for proposing localized policies to improve the ecological environment.

Aim: To report the clinical characteristics, treatment and outcomes of active syphilitic uveitis in human immunodeficiency virus (HIV) positive patients and compare them with the previously published data., Methods: Retrospective analysis of the case series from an infectious disease center in southern China was conducted. Comprehensive review of previously published cases of HIV positive syphilitic uveitis was conducted using the PubMed and Web of Science databases and the references listed in the identified articles., Results: Twelve HIV positive patients with active syphilitic uveitis were collected. All were male, with age of 36.3y (range 27 to 53y). Five (41.7%) had a history of syphilis, and three of them had received anti-syphilis treatment. Ocular manifestations included corneal epithelial defect (13%), complicated cataract (17.4%), vitreous opacity (82.6%), optic disc edema (26.1%), macular edema (30.4%), neuro-retinitis (43.5%), and retinal hemorrhage (26.1%). After standardized syphilitic treatment, intraocular inflammation was reduced and vision improved in all cases. The literature review summarizes 105 previously reported cases of HIV positive syphilitic uveitis. High serum rapid plasma regain (RPR) titers may be associated with severe uveitis and poor vision. Treatment with penicillin, ceftriaxone sodium, or penicillin plus benzylpenicillin instead of using benzylpenicillin alone can significantly improve best-corrected visual acuity (BCVA) in HIV positive ocular syphilis patients., Conclusion: For HIV positive syphilitic uveitis patients, prompt diagnosis and appropriate treatment and follow-up are paramount. In our series, the clinical manifestations are diverse. Syphilis patients treated by penicillin G or long-acting penicillin before may still develop syphilitic uveitis. Patients who relapse after long-term penicillin treatment can still benefit from penicillin G., (International Journal of Ophthalmology Press.)

Under the "Carbon Peak, Carbon Neutral" goal, the systematic evaluation of the carbon emission equivalent (CO 2 eq) and its compositions of the typical A 2 O process has important guiding significance for the low-carbon operation of most municipal sewage plants in China. Based on the operational data on the first municipal sewage plant of Jiaozuo in 2020 and the methods presented in "2019 Refinement to the 2006 IPCC Guidelines for National Greenhouse Gas Inventories, " a systematic evaluation of the CO 2 eq of the typical A 2 O process was established, including direct emissions that were built on the Arrhenius model introducing the water temperature factor and indirect emissions from the three aspects of electricity consumption, agent addition, and sludge transportation. The results showed that the daily emission intensities of CH 4 and N 2 O were (115±56) kg·d -1 and (30±18) kg·d -1 , respectively. Additionally, indirect carbon emissions from electricity consumption and agent addition accounted for 48.4% and 51.3% in the biochemical treatment section, respectively. In 2020, CO 2 eq amounts of total research plant and per unit sewage were 2.17×10 4 t and (0.63±0.07) kg·m -3 , respectively. The magnitude of the proportion of different carbon emission compositions was as follows:sewage electricity (36.5%)>sewage agent (26.6%)>N 2 O direct (15.4%)>sludge agent (9.6%)>sludge electricity (6.7%)>CH 4 direct (4.9%)>sludge transportation (0.3%). System import/export fluxes of carbon and nitrogen elements were calculated, followed by the carbon to nitrogen mass ratio in the sewage plant. Direct carbon emission characteristics of CH 4 and N 2 O and their influencing factors were discussed, respectively. Based on the balance theory of carbon and nitrogen elements in the system, it is proposed that the selective introduction of industrial wastewater may become an important reference measure for the low-carbon operation of municipal sewage plants in the future.

Objective: To investigate the predictive value of SYNTAX-Ⅱ score on long term prognosis of patients diagnosed with chronic total occlusion (CTO) and received percutaneous coronary intervention (PCI). Methods: Patients undergoing CTO-PCI in Fuwai hospital from January 2010 to December 2013 were enrolled in this retrospective analysis. The SYNTAX-Ⅱ score of the patients was calculated. According to SYNTAX-Ⅱ score tertiles, patients were stratified as follows: SYNTAX-Ⅱ≤20, 2027. Primary endpoint was major adverse cardiac events (MACCE), including all-cause death, myocardial infarction, stroke and any revascularization. Secondary endpoints included stent thrombosis, heart failure and target lesion failure (TLF). Patients were followed up by outpatient visit or telephone call at 1 month, 6 months and 1 year after PCI, and annually up to 5 years. Multivariate Cox regression model was used to analyze the independent risk factors of all-cause death in patients undergoing CTO-PCI. The predictive value of SYNTAX score with SYNTAX-Ⅱ score for all-cause death was evaluated by the receiver operating characteristic (ROC) curve and the area under the curve ( AUC ). Results: A total of 2 391 patients with CTO and received PCI were enrolled in this study. The mean age was (57.0±10.5) years, 1 994 (83.40%) patients were male. There were 802 patients in lower tertile group (SYNTAX-Ⅱ≤20), 798 patients in intermediate group (2027). At the end of 5-year follow-up, the loss to follow-up rate of the three groups was 9.10%(73/802), 10.78%(86/798)and 8.85%(70/791), respectively. The rate of all-cause mortality (1.78% (13/729) vs. 3.65% (26/712) vs. 9.02% (65/721), P <0.001), cardiac death (1.37% (10/729) vs. 2.11% (15/712) vs. 4.85% (35/721), P <0.001), target vessel myocardial infarctions (4.25% (31/729) vs. 4.49% (32/712) vs. 7.07% (51/721), P =0.03), probable stent thrombosis (1.51% (11/729) vs. 2.81% (20/712) vs. 3.61% (26/721), P =0.04) and heart failure (1.78% (13/729) vs. 1.97% (14/712) vs. 5.41% (39/721), P <0.001) increased in proportion to increasing SYNTAX-Ⅱ score (all P <0.05). Multivariable Cox regression analysis indicated that female ( HR =2.05, 95% CI 1.12-3.73, P =0.01), left ventricular ejection fraction ( HR =0.97, 95% CI 0.95-1.00, P =0.05) and SYNTAX-Ⅱ score ( HR =1.07, 95% CI 1.02-1.11, P =0.01) were independent predictors for all-cause mortality in patients undergoing CTO-PCI. The predicted value of the SYNTAX-Ⅱ score for all-cause death was significantly higher than the SYNTAX score ( AUC 0.71 vs. 0.60, P =0.003). Conclusion: For CTO patients who underwent percutaneous coronary intervention, SYNTAX-Ⅱ score is an independent predictor for 5-year all-cause death, and SYNTAX-Ⅱ serves as an important predictor for all-cause death in these patients.

Background and Aims: Endoscopic radiofrequency ablation (RFA) is an emerging technique for the palliation of inoperable malignant biliary strictures (MBSs). We aimed to systemically investigate the long-term outcome of RFA in a large cohort of patients., Methods: We recruited 883 patients with various MBSs who underwent endoscopic interventions at two large-volume centers; 124 patients underwent RFA and stenting, whereas 759 underwent stenting alone. To overcome selection bias, we performed 1:4 propensity score matching (PSM). The main outcome was overall survival (OS)., Results: Following PSM, patients in the RFA group showed significantly longer OS (9.5 months; 95% CI: 7.7-11.3 months) than those in the stenting alone group (6.1 months; 95% CI: 5.6-6.6 months; P < .001). In stratified analyses, the improved OS was only demonstrated in the subgroup of extrahepatic cholangiocarcinoma (11.3 months 95% CI: 10.2-12.4 vs 6.9 months 95% CI: 6.0-7.8; P < .001), but not in the subgroups of gallbladder cancer, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, pancreatic cancer, and other metastatic cancers (all P > .05). The survival benefits were noted only in the patients with non-metastatic cholangiocarcinoma (11.5 vs 7.4 months, P < .001)., Conclusions: The survival benefits of endoscopic RFA appear to be limited to patients with extrahepatic cholangiocarcinoma without distant metastasis., (© 2021 Japanese Society of Hepato-Biliary-Pancreatic Surgery.)

Objective To investigate the role of RNA binding protein─upstream-of-N-Ras (UNR) in the development of glioma and its molecular mechanism.Methods First, bioinformatics analysis of CGGA database was performed to detect UNR expression level and prognosis of patients with glioma. Western blot and real-time PCR were used to detect UNR expression level in glioma cell lines and tissues. Next, UNR siRNAs were transfected in glioma cells, and MTS assay and scratch wound-healing assay were used to detect changes in cell proliferation and migration. Then, the candidate UNR target mRNAs were identified by analyzing the sequencing data of UNR iCLIP-seq, RNA sequencing and ribosome profiling databases of human melanoma. RNA immunoprecipitation and biotin pull-down assays were used to identify the UNR target mRNAs in glioma cells. Finally, western blot was used to detect the effect of UNR knockdown on ribosomal protein L9 (RPL9) and RPL9 protein expression level in glioma cell lines. RPL9 siRNA was transfected in A172 and T98G and the expression of vimentin in the cells was detected with western blot.Results Bioinformatics analysis showed that UNR mRNA expression level was significantly higher in high-grade glioma [Grade 2 (n=126), Grade 3 (n=51), Grade 4 (n=128), P<0.001]. UNR high expression levels were associated with poor prognosis (P=0.0177). UNR had high expression level in glioma cell lines and patient samples compared with normal cell lines and normal brain samples (P<0.01). Knockdown of UNR inhibited glioma cells migration (P<0.05), but did not inhibit glioma cells growth in three glioma cell lines. UNR binded the 3' untranslated region (UTR) of PTEN and RPL9 mRNAs. RPL9 protein was significantly highly expressed in most glioma cell lines (n=9) and knockdown of UNR resulted in a downregulation of RPL9 protein expression. Epithelial-mesenchymal transition (EMT)-related marker─vimentin was positively regulated by RPL9.Conclusions UNR could bind to the 3'UTR of PTEN and RPL9 in glioma cell lines, therefore promoting glioma cell migration and regulating the expression of RPL9. Here, we establish a link between UNR and RPL9 protein, which will provide new ideas for the further study of glioma.

Aims: To investigate the association of the Nectin/Necl family genes with the risk of developing NMOSD., Methods: Whole-exome sequencing was performed on two familial NMOSD cases and two unaffected family members. Additionally, 106 patients with sporadic NMOSD and 212 healthy controls (HCs) underwent screening for mutant Necl2. Finally, the molecular weight and cellular localization of mutant NECL2 was examined in transfected HeLa cells., Results: We identified a novel deletion mutation in Necl2 (c.1052&#95;1060delCCACCACCA; p. Thr351&#95;Thr353del), which was associated with disease manifestation in the NMOSD familial cases. The frequency at which the mutation occurred in patients with sporadic NMOSD was significantly higher than for HCs (5.7% and 0, respectively; p<0.01). The mutation was located in the extracellular domain close to the transmembrane region, at a point in the protein sequence characterized by threonine enrichment. The mutant NECL2 had a lower molecular weight and exhibited defective trafficking to the cell surface., Conclusions: Our results suggest that the Necl2 mutation identified herein may be associated with the risk of developing NMOSD. Furthermore, mutated NECL2 may play a role in the pathogenesis of the disease, potentially through its roles in axonal regeneration and/or via neuron-glia interactions that are relevant to myelination., (Copyright © 2017. Published by Elsevier B.V.)

The difference in contents of paclitaxel and 10-deacetylbaccatin III (10-DABIII) in needles between wildtype (WT) and a new cultivar (Zhongdayihao, ZD1) of Taxus yunnanensis was examined. Transcriptome profiling was conducted for different tissues of the ZD1 and WT to illustrate the regulation mechanism of paclitaxel biosynthesis. It was observed that average contents of paclitaxel and 10-DABIII in ZD1 were 4 folds and 32 folds higher than those in WT, respectively. More significant elevations of differential expressed genes (DEGs) from paclitaxel biosynthesis pathway were revealed in ZD1 rather than WT, which should be responsible for the higher contents of paclitaxel and 10-DABIII in the ZD1. Special tissues-dependent expression patterns of paclitaxel biosynthesis DEGs in ZD1 compared to WT were unraveled. The relative higher expressions of paclitaxel biosynthesis genes in needles than other tissues supported the higher content of paclitaxel and 10-DABIII content in needles of ZD1. Attenuation of plant hormone signal transduction pathway led to the lower expression of TFs in ZD1 rather than WT. Besides, the significant negative correlations between differential expressed TFs and DEGs from paclitaxel biosynthesis pathway displayed a possibly negative regulation pattern of these TFs on paclitaxel biosynthesis pathway genes. These results provided new insights into the molecular process of paclitaxel synthesis in Taxus., (Copyright © 2017 Elsevier Masson SAS. All rights reserved.)

Polycomb group (PcG) proteins play an important role in development and stem cell maintenance, and their dysregulation have been closely linked to oncogenesis and cancer stem cell phenotypes. Here, we found that nervous system polycomb 1 (NSPc1) was highly expressed in stem cell-like glioma cells (SLCs). Knockdown of NSPc1 in SLCs resulted in impaired neurosphere formation and self-renewal abilities, down-regulated expression of stemness markers such as NESTIN, CD133 and SOX2, and decreased capacity to propagate subcutaneous xenografts. In contrast, glioma cells overexpressing NSPc1 exhibited a stem cell-like phenotype, up-regulated expression of stemness markers NESTIN, CD133 and SOX2, and an enhanced capacity to propagate subcutaneous xenografts. Furthermore, we identified that NSPc1 epigenetically repressed the expression of retinol dehydrogenase 16 (RDH16) by directly binding to a region upstream (-1073 to -823) of the RDH16 promoter. Next, we confirmed that RDH16 is a stemness suppressor that partially rescues SLCs from the NSPc1-induced increase in neurosphere formation. Finally, we showed that ATRA partly reversed the NSPc1-induced stemness enhancement in SLCs, through mechanisms correlated with an ATRA-dependent decrease in the expression of NSPc1. Thus, our results demonstrate that NSPc1 promotes cancer stem cell self-renewal by repressing the synthesis of ATRA via targeting RDH16 and may provide novel targets for glioma treatment in the future.

In plants, microRNAs (miRNAs) play regulatory roles in response to various environmental stresses. In order to illustrate the regulation mechanisms of miRNAs involving the different Cd accumulation abilities between a low-shoot-Cd cultivar (QLQ) and a high-shoot-Cd cultivar (T308) of water spinach (Ipomoea aquatic Forsk.), six sRNA libraries at 3 different time points were constructed. Only 5 miRNAs were exclusively regulated in QLQ, among them, miRNA395 was up-regulated, which was supposed to enhance the Cd retention and detoxification in root. Also, the alterations of miRNA5139, miRNA1511 and miRNA8155 contributed to the attenuation of Cd translocation into the shoot of QLQ. More differentially expressed miRNAs were observed in T308, indicating more complex response was adopted by T308 under Cd stress. miRNA397 exclusively regulated in T308 has enhanced the Cd influx of T308 under Cd treatments. Besides, the Cd translocation of T308 was strengthened due to the up-regulation of MATE efflux family, which was targeted by miRNA3627. Our results unraveled the effects of the cultivar-dependent expression of these specific miRNAs on the different Cd accumulation and translocation abilities of QLQ and T308. These findings provide a new perspective for the molecular assisted breeding of low-Cd cultivars for leaf-vegetables., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Hair is a promising, non-invasive, human biomonitoring matrix that can provide insight into retrospective and integral exposure to organic pollutants. In the present study, we measured the concentrations of organophosphate flame retardants (PFRs) in hair and serum samples from university students in Guangzhou, China, and compared the PFR concentrations in the female hair segments using paired distal (5~10cm from the root) and proximal (0~5cm from the root) samples. PFRs were not detected in the serum samples. All PFRs except tricresyl phosphate (TMPP) and tri-n-propyl phosphate (TPP) were detected in more than half of all hair samples. The concentrations of total PFRs varied from 10.1 to 604ng/g, with a median of 148ng/g. Tris(chloroisopropyl) phosphate (TCIPP) and tri(2-ethylexyl) phosphate (TEHP) were the predominant PFRs in hair. The concentrations of most PFRs in the distal segments were 1.5~8.6 times higher than those in the proximal segments of the hair (t-test, p<0.05), which may be due to the longer exposure time of the distal segments to external sources. The values of log (PFR concentrations-distal/PFR concentrations-proximal) were positively and significantly correlated with log KOA of PFRs (p<0.05, r=0.68), indicating that PFRs with a higher log KOA tend to accumulate in hair at a higher rate than PFRs with a lower log KOA. Using combined segments of female hair, significantly higher PFR concentrations were observed in female hair than in male hair. In contrast, female hair exhibited significantly lower PFR concentrations than male hair when using the same hair position for both genders (0-5cm from the scalp). The controversial results regarding gender differences in PFRs in hair highlight the importance of segmental analysis when using hair as an indicator of human exposure to PFRs., (Copyright © 2016 Elsevier Inc. All rights reserved.)

A low-shoot-Cd (QLQ) and a high-shoot-Cd cultivar (T308) of water spinach (Ipomoea aquatica Forsk.) were used to investigate molecular mechanism of the genotype difference in cadmium (Cd) accumulation. RNA-Seq under 9 and 72 h cadmium exposures (5 mg L(-1)) were undertaken to explore Cd induced genotype differences in molecular processes. In total, 253 747 540 clean reads were assembled into 57 524 unigenes. Among them, 6136 and 10 064 unigenes were differentially expressed in QLQ and T308, respectively. Cell wall biosynthesis genes, such as GAUT and laccase, and three Cd efflux genes (Nramp5, MATE9, and YSL7) had higher expression levels in QLQ, while the genes in sulfur and glutathione metabolism pathway, e.g., sulfate transporter and cysteine synthase, showed higher expression levels in T308. These findings would be useful for further understanding of the mechanisms related to genotype-dependent Cd accumulation and developing the molecular assisted screening and breeding of low-shoot-Cd cultivars for water spinach.

To reduce cadmium (Cd) pollution of food chains, screening and breeding of low-Cd-accumulating cultivars are the focus of much study. Two previously identified genotypes, a low-Cd-accumulating genotype (LAJK) and a high-Cd-accumulating genotype (HAJS) of pakchoi (Brassica chinesis L.), were stressed by Cd (12.5 μM) for 0 h (T0), 3 h (T3) and 24 h (T24). By comparative transcriptome analysis for root tissue, 3005 and 4343 differentially expressed genes (DEGs) were identified in LAJK at T3 (vs T0) and T24 (vs T3), respectively, whereas 8677 and 5081 DEGs were detected in HAJS. Gene expression pattern analysis suggested a delay of Cd responded transcriptional changes in LAJK compared to HAJS. DEG functional enrichments proposed genotype-specific biological processes coped with Cd stress. Cell wall biosynthesis and glutathione (GSH) metabolism were found to involve in Cd resistance in HAJS, whereas DNA repair and abscisic acid (ABA) signal transduction pathways played important roles in LAJK. Furthermore, the genes participating in Cd efflux such as PDR8 were overexpressed in LAJK, whereas those responsible for Cd transport such as YSL1 were more enhanced in HAJS, exhibiting different Cd transport processes between two genotypes. These novel findings should be useful for molecular assisted screening and breeding of low-Cd-accumulating genotypes for pakchoi.

Organophosphorus flame retardants (OPFRs) are important alternatives to brominated flame retardants (BFRs), but information on their contamination of the environment in China is rare. We examined the occurrence of 12 OPFRs in indoor dust in four microenvironments of southern China, including a rural electronic waste (e-waste) recycling area, a rural non-e-waste area, urban homes, and urban college dormitory rooms. The OPFR concentrations (with a median of 25.0 μg g(-1)) were highest in the e-waste area, and the concentrations in other three areas were lower and comparable (7.48-11.0 μg g(-1)). The levels of OPFRs in the present study were generally relatively lower than the levels of OPFRs found in Europe, Canada, and Japan because BFRs are still widely used as the major FRs in China. The composition profile of OPFRs in the e-waste area was dominated by tricresyl phosphate (TCP) (accounting for 40.7%, on average), while tris(2-chloroethyl) phosphate (TCEP) was the most abundant OPFR (64.4%) in the urban areas (homes and college dormitories). These two distribution patterns represent two OPFR sources (i.e., emissions from past e-waste and from current household products and building materials). The difference in the OPFR profiles in the rural area relative to the OPFR profiles in the urban and e-waste areas suggests that the occurrence of OPFRs is due mainly to emissions from characteristic household products in rural homes. Although human exposures to all the OPFRs were under the reference doses, the health risk for residents in the e-waste area is a concern, considering the poor sanitary conditions in this area and exposure from other sources., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Concentrations of heavy metals (Cd, Pb, Cu, Zn, and Ni) were measured in the foodstuffs, house dust, underground/drinking water, and soil from an electronic waste (e-waste) area in South China. Elevated concentrations of these potentially toxic metals were observed in the samples but not in drinking water. The health risks for metal exposure via food consumption, dust ingestion, and drinking water were evaluated for local residents. For the average residents in the e-waste area, the non-carcinogenic risks arise predominantly from rice (hazard index=3.3), vegetables (2.2), and house dust (1.9) for adults, while the risks for young children are dominated by house dust (15). Drinking water may provide a negligible contribution to risk. However, local residents who use groundwater as a water supply source are at high non-carcinogenic risk. The potential cancer risks from oral intake of Pb are 8×10(-5) and 3×10(-4) for average adults and children, and thus groundwater would have a great potential to induce cancer (5×10(-4) and 1×10(-3)) in a highly exposed population. The results also reveal that the risk from oral exposure is much higher than the risk from inhalation and dermal contact with house dust., (Copyright © 2013 Elsevier Inc. All rights reserved.)

To test the effect of adding different monosaccharide groups to a non-phloem-mobile insecticide on the phloem mobility of the insecticide, a series of conjugates of different monosaccharides and fipronil were synthesized using the trichloroacetimidate method. Phloem mobility tests in castor bean ( Ricinus communis L.) seedlings indicated that the phloem mobility of these conjugates varied markedly. L-Rhamnose-fipronil and D-fucose-fipronil displayed the highest phloem mobility among all of the tested conjugates. Conjugating hexose, pentose, or deoxysugar to fipronil through an O-glycosidic linkage can confer phloem mobility to fipronil in R. communis L. effectively, while the -OH orientation of the monosaccharide substantially affected the phloem mobility of the conjugates.

Human hair and indoor dust from urban, e-waste, and rural areas in south China were collected and analyzed for brominated flame retardants (BFRs). BFRs concentrations in hair from occupational e-waste recycling workers were higher than those from non-occupational exposed residents in other sampling areas. Polybrominated diphenyl ethers (PBDEs) and decabromodiphenyl ethane (DBDPE) are two major BFRs in hair samples. The PBDE congener profiles in hair from the e-waste area are different from those from urban and rural areas with relatively higher contribution of lower brominated congeners. DBDPE, instead of BDE209, has become the major BFR in non-e-waste recycling areas. Significant correlations were found between hair level and dust level for DBDPE and BTBPE but not for PBDEs. The different PBDE congener profiles between dust and hair may suggest that exogenous exposure to the PBDE adsorbed on dust is not a major source of hair PBDEs., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Heavy metals were measured in hair from occupationally and nonoccupationally exposed populations in an e-waste recycling area and from residents from a control rural town. The levels of five heavy metals were in the following order of Zn > Pb, Cu > Cd > Ni, with the highest levels found in the occupationally exposed workers. The levels of Cd, Pb, and Cu were significantly higher in residents from the e-waste recycling area than in the control area. Elevated Cd, Pb, and Cu contents along with significant positive correlations between them in hair from the e-waste recycling area indicated that these metals were likely to have originated from the e-waste recycling activities. The similarity in heavy metal pattern between children and occupationally exposed workers indicated that children are particularly vulnerable to heavy metal pollution caused by e-waste recycling activities. The increased Cu exposure might be a benefit for the insufficient intake of Cu in the studied area. However, the elevated hair Cd and Pb levels implied that the residents in the e-waste area might be at high risk of toxic metal, especially for children and occupationally exposed workers.

Dechlorane Plus (DP) and a dechlorination product, 1,6,7,8,9,14,15,16,17,17,18-octadeca-7,15-diene (anti-Cl(11)-DP), were measured in human hair and indoor dust collected from an e-waste recycling area and two control areas (rural and urban) in South China. DP was detected in hair and dust samples at concentrations ranging from 0.02-58.32 ng/g and 2.78-4197 ng/g, respectively. anti-Cl(11)-DP, mainly detected in human hair and dust samples from the e-waste recycling area, ranged from nd (nondetected) to 0.23 ng/g in hair and from nd to 20.22 ng/g in dust. Average values of anti-DP fractional abundance (f(anti) ratio) in hair of e-waste dismantling workers (0.55 ± 0.11) and dust from e-waste recycling workshops (0.54 ± 0.15) were significantly lower than those in other groups (0.62-0.76 means for hair and 0.66-0.76 means for dust). Significantly positive correlation between DP concentrations in dust and hair and similarity in f(anti) ratios between hair and dust suggest that ingestion of dust comprise one of the major routes for DP exposure. Significantly positive relationships were also observed between anti-Cl(11)-DP and anti-DP for both hair and dust samples with similar regression line slopes. The ratios of anti-Cl(11)-DP to anti-DP between hair and dust show no significant difference. These results suggest that anti-Cl(11)-DP in the human body is likely accumulated from the environmental matrix and not formed from biotransformation of the parent DP.

Background: The downstream of tyrosine kinase/docking protein (Dok) adaptor protein family has seven members, Dok1 to Dok7, that act as substrates of multiple receptor tyrosine kinase and non-receptor tyrosine kinase. The tropomyosin-related kinase (Trk) receptor family, which has three members (TrkA, TrkB and TrkC), are receptor tyrosine kinases that play pivotal roles in many stages of nervous system development, such as differentiation, migration, axon and dendrite projection and neuron patterning. Upon related neurotrophin growth factor stimulation, dimerisation and autophosphorylation of Trk receptors can occur, recruiting adaptor proteins to mediate signal transduction., Results: In this report, by using yeast two-hybrid assays, glutathione S-transferase (GST) precipitation assays and coimmunoprecipitation (Co-IP) experiments, we demonstrate that Dok6 selectively binds to the NPQY motif of TrkC through its phosphotyrosine-binding (PTB) domain in a kinase activity-dependent manner. We further confirmed their interaction by coimmunoprecipitation and colocalisation in E18.5 mouse cortex neurons, which provided more in vivo evidence. Next, we demonstrated that Dok6 is involved in neurite outgrowth in mouse cortex neurons via the RNAi method. Knockdown of Dok6 decreased neurite outgrowth in cortical neurons upon neurotrophin 3 (NT-3) stimulation., Conclusions: We conclude that Dok6 interacts with the NPQY motif of the TrkC receptor through its PTB domain in a kinase activity-dependent manner, and works as a novel substrate of the TrkC receptor involved in NT-3-mediated neurite outgrowth in mouse cortex neurons.

The area we studied is Lake Yamzho Yumco Basin (28 degrees 27'-29 degrees 12'N, 90 degrees 08'-91 degrees 45'E), the largest inland lake basin in southern Tibetan Plateau, China. Using the SPOT-VGT NDVI vegetation index from 1998 to 2007 in the basin, the temporal and spatial variation characteristics of NDVI and its correlation with the major climatic factors (air temperature, precipitation) were analyzed. The results show that the average NDVI of the lake basin ranges from 0.12 to 0.31 and its seasonal change is obvious; the NDVI begins to rise rapidly in May and reaches the maximum value in early September. The average NDVI of the basin shows the slow increasing trend during 1998 to 2007, and it indicates that the eco-environment of the basin is recovering. The high value of NDVI has close relationships with water supply, altitude and vegetation types, so NDVI is relatively high near water sources and is the highest in meadow grassland. The summer air temperature and precipitation are the important climate elements that influence the vegetation in the basin, and the linear correlation coefficients between NDVI and air temperature and precipitation are 0.7 and 0.71, respectively. In recent years, warm and humid trend of the local climate is prevailing to improve the ecological environment in Yamzho Yumco Basin.

Objective: To explore the influences of the restoration of neural adhesion molecule NECL1 on the morphology, migration, and invasion of NECL1-deficient glioma cell lines., Methods: Scratch and Transwell assays were used to observe the cell migration and invasion, the activities of extracellular metalloproteinases were measured, and the cell morphology was observed. Astrocytes marker glial fibrillary acidic protein was detected by Western blot after the restoration of NECL1 in glioma U251 cell line., Results: In NECL1-deficient U251 glioma cell lines, migration and invasion were inhibited. The U251 cells was differentiated potentially to astrocytes, and glial fibrillary acidic protein was up-regulated after the restoration of the NECL1 expression., Conclusion: As a potential tumor repressor, the neural adhesion molecule NECL1 can inhibit the migration and invasion of glioma cell and induces its differentiation.

Background: MicroRNAs (miRNAs) are small noncoding regulatory RNAs whose aberrant expression may be observed in many malignancies. However, few data are yet available on human primary medulloblastomas. This work aimed to identify that whether miRNAs would be aberrantly expressed in tumor tissues compared with non-tumorous cerebellum tissues from same patients, and to explore a possible role during carcinogenesis., Methods: A high throughput microRNA microarray was performed in human primary medulloblastoma specimens to investigate differentially expressed miRNAs, and some miRNAs were validated using real-time quantitative RT-PCR method. In addition, the predicted target genes for the most significantly down- or up-regulated miRNAs were analyzed by using a newly modified ensemble algorithm., Results: Nine miRNA species were differentially expressed in medulloblastoma specimens versus normal non-tumorous cerebellum tissues. Of these, 4 were over expressed and 5 were under expressed. The changes ranged from 0.02-fold to 6.61-fold. These findings were confirmed using real-time quantitative RT-PCR for most significant deregulated miRNAs (miR-17, miR-100, miR-106b, and miR-218) which are novel and have not been previously published. Interestingly, most of the predicted target genes for these miRNAs were involved in medulloblastoma carcinogenesis., Conclusions: MiRNAs are differentially expressed between human medulloblastoma and non-tumorous cerebellum tissue. MiRNAs may play a role in the tumorigenesis of medulloblastoma and maybe serve as potential targets for novel therapeutic strategies in future.

Phytochelatins (PCs) play an important role in detoxification of heavy metals in plants. PCs are synthesized from glutathione by phytochelatin synthase (PCS), a dipeptidyltransferase. Sesbania rostrata is a tropical legume plant that can tolerate high concentrations of Cd and Zn. In this study, the S. rostrata PCS gene (SrPCS) and cDNAs were isolated and characterized. Southern blot and sequence analysis revealed that a single copy of the SrPCS gene occurs in the S. rostrata genome, and produces four different SrPCS mRNAs and proteins, SrPCS1-SrPCS4, by alternative splicing of the SrPCS pre-mRNA. The SrPCS1 and SrPCS3 proteins conferred Cd tolerance when expressed in yeast cells, whereas the SrPCS2 and SrPCS4 proteins, which lack the catalytic triad and the N-terminal domains, did not. These results suggested that SrPCS1 and SrPCS3 have potential applications in genetic engineering of plants for enhancing heavy metal tolerance and phytoremediation of contaminated soils.

Atmospheric correction of remote sensing image is the premise of quantitative remote sensing. The present paper evaluated the capability of FLAASH (fast line-of-sight atmospheric analysis of spectral hypercubes)in ENVI software to make atmospheric correction for EO-1 Hyperion hyperspectral image. Hyperion hyperspecreal image of Zhangye city in Heihe River valley of Gansu province, China was acquired on September 10, 2007. Canopy spectra, biochemical component and GPS data of 41 plots were measured in near real-time during the satellite overpass. Hyperion hyperspectral image was geometrically corrected using Lansat-7 ETM+ image, then DN values were transformed to radiance and apparent reflectance, and atmospheric correction of Hyperion image was made using FLAASH. The resulting radiance, apparent reflectance and reflectance after FLAASH of four typical objects, including corn, water body, desert and building, were compared. ASD spectra of corn were resampled to Hyperion corresponding bands using Gaussian filter function. The comparison between ASD resampled spectra and Hyperion spectra after FLAASH demonstrated that the atmospheric correction using FLAASH is very effective and these two spectra are consistent with each other and the correlation coefficient reached 0.987.

Human DIXDC1 is a member of Dishevelled-Axin (DIX) domain containing gene family which plays important roles in Wnt signaling and neural development. In this report, we first confirmed that expression of Ccd1, a mouse homologous gene of DIXDC1, was up-regulated in embryonic developing nervous system. Further studies showed that Ccd1 was expressed specifically in neurons and colocalized with early neuronal marker Tuj1. During the aggregation induced by RA and neuronal differentiation of embryonic carcinoma P19 cells, expressions of Ccd1 as well as Wnt-1 and N-cadherin were dramatically increased. Stable overexpression of DIXDC1 in P19 cells promoted the neuronal differentiation. P19 cells overexpressing DIXDC1 but not the control P19 cells could differentiate into Tuj1 positive cells with RA induction for only 2 days. Meanwhile, we also found that overexpression of DIXDC1 facilitated the expression of Wnt1 and bHLHs during aggregation and differentiation, respectively, while inhibited gliogenesis by down-regulating the expression of GFAP in P19 cells. Thus, our finding suggested that DIXDC1 might play an important role during neurogenesis, overexpression of DIXDC1 in embryonic carcinoma P19 cells promoted neuronal differentiation, and inhibited gliogenesis induced by retinoic acid.

Objective: To provide a set of useful analysis tools for the researchers to explore the microRNA data., Methods: The R language was used for generating the Graphical Users Interface and implementing most functions. Some Practical Extraction and Report Language (Perl) scripts were used for parsing source files., Results: We developed a graphical R package named miRE, which was designated for the analysis of microRNA functions, genomic organization, etc. This package provided effective and convenient tools for molecular biologists to deal with routine analyses in microRNA-related research. With its help, the users would be able to build a desktop-centered microRNA research environment quite easily and effectively. miRE is freely available at http://www. biosino.org/-kanghu/WorkPresentation/miRE/miRE.html. A detailed user manual and tutorials with example code and image are also available., Conclusion: miRE is a tool providing an open-source, user-friendly, integrated interface for microRNA-related analysis. With its help, researchers can perform microRNA-related analysis more efficiently.

Objective: To study the expression pattern of Polycomb gene Nspc1 at the early developmental stage in zebrafish., Methods: In situ hybridization probe for Nspc1 was designed according to the GenBank information. Collecting zebrafish embryos at different stages including one cell stage, two-cell stage, bud stage, and somites stage, we hybridized them with the prepared probe. Then the hybridization signals at different intervals were observed and photographed at the right time., Results: Nspc1 was expressed globally at the early stage. Its expression specificity began at the somites stage, mainly in the nervous system of the head., Conclusion: Nspc1 may play essential roles in the early stage development of zebrafish, especially in the nervous system.

Objective: To study the regulation role of tumor suppressor NECL1 on the proliferation of glioma cell line., Methods: We detected the expression level of NECL1 in human normal brain tissue and glioma cell lines using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. T98G cell line in which NECL1 was silent and whose transfection efficiency was relatively high as target cell was chosen, and the effect of NECL1 on the proliferation of T98G cell line in vitro was detected by using cell growth curve, flow cytometry, and Hoechst staining., Results: NECL1 was abrogated or markedly reduced in 6 glioma cell lines. When NECL1 was overexpressed in T98G cell line, the cell growth rate obviously decreased and the number of apoptotic cells remarkably increased when compared with the control group., Conclusion: NECL1 may inhibit the proliferation of T98G cells by inducing its apoptosis.

Objective: To detect the expression of CBX7 in human glioma and investigate the potential regulatory effect of abnormally expressed microRNAs on CBX7 expression., Methods: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression pattern of CBX7 in 2 human normal brain tissues, 9 glioma tissues, and 3 glioma cell lines. Miranda algorithm and Ensemble Machine Learning algorithm were combined to predict miRNAs that target human CBX7. The expression of miR-9 in those tissues and cell lines were detected by real-time PCR. After miR-9 overexpression in 293ET and miR-9 knock-down in T98G, luciferase assay and Western blot were used to confirm the effect of miR-9 on CBX7 expression. MTT assay and flow cytometry were applied to detect the effect of miR-9 knock-down on T98G cells., Results: No obvious difference in the CBX7 mRNA level between normal and tumor tissues was observed, while the protein level of CBX7 was abrogated or markedly reduced in glioma tissues and cell lines. Several miRNAs including miR-9 may target CBX7 by bioinformatics prediction. MiR-9 was up-regulated in glioma tissues and cell lines. In 293ET cell, luciferase activity of CBX7-3'UTR reporter was decreased to 24% after miR-9 overexpression. After miR-9 knock-down in T98G cell, the luciferase activity was increased by 1.8 fold and there was no change of CBX7 mRNA, while the protein level of endogenous CBX7 was significantly increased. The number of survival T98G cells increased and cells in G1 phase decreased after miR-9 knock-down., Conclusion: In human glioma, CBX7 is down-regulated by the inhibition of miR-9 at posttranscriptional level.

Objective: To study the role of cell adhesion molecules Necl1 in synaptogenesis in primary cultured neurons., Methods: Semi-quantitive reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression pattern of Necl1 in the neuronal differentiation cell model in vitro. Western blot was performed to detect the expression pattern of Necl1 in primary cultured rat neurons and in purified synaptosome. Immunofluoresence was used to detect the synapse formation in primary neurons and in 293 cells co-culture and to detect the density of synapses in primary neuron with ectopic expression of Necl1., Results: Necl1 expression increased after retinoic acid (RA) induction in SH-SY5Y and P19 cells. The increase of Necl1 expression was consistent with the days of primary neurons culture in vitro, and Necl1 partly localized in synaptosome. The overexpression of Necl1 in 293 cells induced the synapse formation between cocultured 293 cells and neurons. Ectopic expression of Necl1 in primary neurons increased the density of synapses., Conclusion: Necl1 plays an important role in neuronal synapse formation.

Objective: To generate a sensitive tool for noninvasive monitoring of a therapeutic gene vasostatin., Methods: We fused the bioluminescent reporter gene firefly luciferase to the therapeutic transgene vasostatin and ensured that these two proteins would not interrupt each other and kept their own natural character., Results: We therefore examined clones of PC3 cells stably expressing fusion gene and positive controlfluc with bioluminescence. In vivo imaging of PC3-Fluc subcutaneous tumors showed that the mean tumor bioluminescence increased in animals over several weeks., Conclusion: Noninvasive monitoring facilitates the detection of gene expression in vivo and in vitro.

Calcyclin-binding protein (Siah-1-Interacting Protein, CacyBP/SIP), is a calcium signaling protein involved in the degradation of beta-catenin, however, little is known about its role in reproductive biology. The present study was to character its temporospatial expression pattern and regulation in mouse uterus and to investigate whether it plays a role in the regulation of normal endometrial events. While prominently expressed in both luminal and glandular epithelia, CacyBP underwent dynamic changes during early pregnancy. CacyBP expression was observed weakly from days 1-4. An intense accumulation in luminal and glandular epithelia as well as decidua surrounding the embryo at later stages (days 5-7) was observed. Most notably, CacyBP accumulation in trophoblast was pronounced at day 7. Using ovariectomized and pseudopregnant mice, we found that progesterone (P(4)) and 17beta-estradiol (E(2)) led to increased expression of CacyBP gene and this could be abolished by Ru486 and tamoxifen, respectively. Antisense oligonucleotides (ODNs) against CacyBP significantly inhibited cultured endometrial stromal cells' (ESCs) apoptosis induced by UV irradiation. Injection of antisense ODNs into mouse uterine horn severely impaired the number of implanted blastocysts. Taken together, our results suggested that CacyBP expression was positively regulated by P(4) and E(2). CacyBP may be involved in the regulation of endometrial cell apoptosis during early pregnancy and play an important role in mouse endometrial events such as pregrancy establishment.

Objective: To express and purify the recombinant human pigment epithelium-derived factor (PEDF) which inhibits the proliferation of the endothelium cells from blood vessel in E.coli., Methods: PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT., Results: The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC., Conclusion: The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression.

Objective: To study the distribution of collagen IX gene in the disc and to determine its role in the pathogeny of idiopathic scoliosis (IS)., Methods: The data included apical disc and intermediate disc from 14 cases of adolescent IS, 26 discs from 13 cases of scoliosis of confirmed pathogeny (CPS), which included 10 cases of congenital scoliosis and neurofibromatosis scoliosis. Six discs were obtained from 3 cases of normal young man served as controls. The distribution of collagen IX was studied in the apical disc of IS by immunohistochemistry and in situ hybridization (ISH) with RNA probe. The figure of collagen IX hybridization in the endplate cartilage was input to the figure analysis system. The mRNA content of collagen IX was compared between each group by SPSS software., Results: Collagen IX was mainly distributed in the inner fibrous annulus, nucleus and endplate cartilage. Collagen IX was secreted by the little round chondrocyte-like cells, which was not expressed in the hypertrophic cells. There was significant difference of collagen IX mRNA content between the concave side of apical disc in the IS and the normal disc(P < 0.05), and also between intermediate vertebrae of CS group and normal., Conclusions: There is no obvious abnormal distribution of collagen IX in the disc of idiopathic scoliosis. Collagen IX may be related to the pathogensis of IS. More investigation such as quantity analysis and protein function determination is needed to confirm its role in the pathogenicity of IS.

We have previously characterized transcription factor LZIP to be a growth suppressor targeted by hepatitis C virus oncoprotein. In search of proteins closely related to LZIP, we have identified a liver-enriched transcription factor CREB-H. LZIP and CREB-H represent a new subfamily of bZIP factors. CREB-H activates transcription by binding to cAMP responsive element, box B, and ATF6-binding element. Interestingly, CREB-H has a putative transmembrane (TM) domain and it localizes ambiently to the endoplasmic reticulum. Proteolytic cleavage that removes the TM domain leads to nuclear translocation and activation of CREB-H. CREB-H activates the promoter of hepatic gluconeogenic enzyme phosphoenolpyruvate carboxykinase. This activation can be further stimulated by cAMP and protein kinase A. CREB-H transcript is exclusively abundant in adult liver. In contrast, the expression of CREB-H mRNA is aberrantly reduced in hepatoma tissues and cells. The enforced expression of CREB-H suppresses the proliferation of cultured hepatoma cells. Taken together, our findings suggest that the liver-enriched bZIP transcription factor CREB-H is a growth suppressor that plays a role in hepatic physiology and pathology.

Objective: To study the distribution of collagen X in the intervertebral disc and determine its role in the pathogeny of idiopathic scoliosis (IS)., Methods: The data included apical disc and intermediate disc from 14 cases of AIS, 26 discs from 13 cases of scoliosis of confirmed pathogeny which included 10 cases of congenital scoliosis and neurofibromatosis scoliosis (CS group). Six discs were obtained from 3 cases of sudden death of normal young man served as controls. The distribution of collagen X in the apical disc of IS was examined by immunohistochemistry and in situ hybridization (ISH) with RNA probe. The figure of collagen X hybridization in the endplate cartilage was input to the figure analysis system. The mRNA content of collagen X was compared between every 2 groups by SPSS software., Results: Collagen X was mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its mRNA was expressed in the hypertrophic chondrocyte. Positive signal was found in the nuclei of several scoliosis patients, which was not related to the pathogeny. The collagen X mRNA contents of the apical disc and intermediated disc of the IS group and of the CS group were significantly higher than those of the normal group (P < 0.05). There was no difference between the IS and CS groups., Conclusions: Collagen X is mainly distributed around the hypertrophic chondrocyte in the endplate cartilage. Its can also be secreted by the chondrocyte-like cells in the nucleus under special condition. Higher expression of collagn X gene in scoliosis patients may be the effects of long term abnormal stress which causes calcification of endplate cartilage. Collagen X expressed in the nucleus may be the result of secretion of chondrocyte-like cells in the disc under abnormal mechanical condition.

Responses of Sesbania rostrata and S. cannabina to Pb, Zn, Cu and Cd toxicities were assessed by a seed-suspending seedbed(SSS) approach. The results showed that the SSS approach was suitable for testing the tolerance of a plant to the stress of toxic metals. The endpoints include seed germination success, straightened radicle and hypocotyl of the seedlings from the seeds. The measurements could be done easily and accurately. It was found that the elongation of radicle was the most sensitive indicator to the stress of heavy metals among the endpoints. When exposure to lower or medium concentrations of Pb, Zn, and Cd, the development of the lateral roots were favorable. Species of S. rostrata was more tolerant than S. cannabina to the heavy metals, especially to Zn and Cd. The ED50 of Pb, Zn, Cu and Cd were 32.90, 5.32, 4.40 and 12.00 microg/ml for S. rostrata, respectively, and they were 30.11, 2.87, 4.05 and 4.94 microg/ml respectively for S. cannabina.

Background: Total graft excision with in situ or extra-anatomic revascularization is considered mandatory to treat infection involving the body of aortic grafts. We present a series of nine patients with this complication and such severe comorbid medical illnesses or markedly hostile abdomens that traditional treatments were precluded. In these patients selective complete or partial graft preservation was used., Methods: Over the past 20 years we have treated nine infected infrarenal aortic prosthetic grafts with complete or partial graft preservation, because excision of the graft body was not feasible. In all nine patients infection of the main body of the aortic graft was documented at computed tomography or surgery. Essential adjuncts included percutaneous or operative drain placement into retroperitoneal abscess cavities and along the graft, with instillation of antibiotics three times daily, repeated debridement of infected groin wounds, and intravenous antibiotic therapy for at least 6 weeks., Results: One patient with purulent groin drainage treated with complete graft preservation died of sepsis. One patient with groin infection treated with complete graft preservation initially did well, but ultimately required total graft excision 5 months later, after clinical improvement. In four patients complete graft preservation was successful; two patients required excision of an occluded infected limb of the graft; and one patient underwent subtotal graft excision, leaving a graft remnant on the aorta, and axillopopliteal bypass. In summary, seven of nine patients survived hospitalization after complete or partial graft preservation; amputation was avoided in all but one patient; and no recurrent infection developed over mean follow-up of 7.6 years (range, 2-15 years)., Conclusions: Although contrary to conventional concepts, partial or complete graft preservation combined with aggressive drainage and groin wound debridement is an acceptable option for treatment of infection involving an entire aortic graft in selected patients with prohibitive risks for total graft excision. This treatment may be compatible with long-term survival and protracted absence of signs or symptoms of infection.

Objective: To screen out the specific antibody clones against amyloid beta peptide 40, and clone the antibody gene and express it in a bacterial system, so as to provide a solid basis for novel diagnostic and therapeutic methods for Alzheimer's Disease., Methods: beta amyloid peptide 40 was bound on the solid surface of Nunc plates as antigen to screen the binding clones from a phage-display human single-chain Fv antibody library. After five rounds of bio-panning, the host E. coli TG1 was infected with eluted filamentous phage from the last turn of selection. 55 well-separated colonies were picked randomly from the plates and the specific positive clones were identified by ELISA test. The single-chain Fv antibody gene was sequenced and their amino acids sequence was deduced. The scFv antibody gene was sub-cloned into a protokayotic expression vector pGEX-6P-1 and transformed into bacteria strain BL21 to express the glutathione-S-transferase (GST) fusion single-chain antibody., Results: ELISA test showed that 33 of the 55 clones could bind amyloid beta peptide 40 and 10 of the 33 clones could be inhibited by amyloid beta peptide 40 itself to below 50% of its original binding activities. Five of the 10 clones could also be inhibited by amyloid beta peptide 1-16 to the same level, which meant that the binding epitope of the antibody from the 5 clones was between first to sixteenth amino acids at amino-end of amyloid beta peptide 40. DNA sequencing data demonstrated that the gene of the single-chain antibody specifically against amyloid beta peptide 40 was consisted of 768 bp and the deduced amino acids sequence confirmed its typical antibody structure. The complement determinant regions and framework regions were discriminated empirically. After cloning the antibody gene into a protokayotic system, the GST fusion antibody was expressed as the expected size., Conclusions: After five rounds of bio-panning and subsequently serial ELISA testing, the specific antibody clones against amyloid beta peptide 40 were screened out successfully. The antibody gene DNA sequence and amino acids sequence were analyzed and confirmed. The fusion antibody was expressed as expected in the bacterial system.

Objective: To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain., Methods: According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA., Results: Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained., Conclusions: The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.

Objective: To get the cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain., Methods: Using the SARS-CoV PUMC2 strain genomic RNA as the template, the cDNA fragments were amplified by RT-PCR, the PCR products were further purified and ligated into the pGEM-T vector, and all the clones obtained were sequenced., Results: The cDNA clones which cover the whole genome of SARS-CoV PUMC2 strain were obtained., Conclusions: These cDNAs can be provided for the function study of SARS-CoV proteins and the construction of full-length infectious cDNA clone of SARS-CoV.

Objective: To identify and clone the gene encoding human TNFLP (tumor necrosis factor like protein) for some functional study on TNFLP., Methods: The full-length cDNA of TNFLP was isolated from fetal brain cDNA library. Several kinds of software were used to analyze nucleotide sequence and amino acid sequence of TNFLP. TNFLP mRNA distribution was identified by Northern blot. TNFLP-C and TNFLP-N were expressed in E. coli with GST expression system., Results: The cDNA of human TNFLP was 2,112 bp, which encoded protein of 208 amino acid. Hydrophobility analysis found there were two hydrophobility regions of human TNFLP. TNFLP-C (112-207 amino acid) and mouse TNF-alpha were homologous. The identity of their amino acid sequence was 42%. Moreover, both of them had a motif-TYKRL. TNFLP was located in chromosome 16. Human TNFLP was widely expressed in various human tissues. Northern blot showed TNFLP was highly expressed in heart, brain and spleen, only one transcript can be seen. GST-TNFLP-C and GST-TNFLP-N fusion proteins were obtained., Conclusions: Tissue expression spectrum of TNFLP and prokaryotic expression of TNFLP have been done, which establish the base for the functional analysis of TNFLP.

Objective: To prokaryoticly express and purify HuD protein and its RNA recognition motifs., Methods: HuD protein was prokaryoticly expressed and purified by molecular cloning technology. Its biologic activity was testified by Western Blot., Results: Purified HuD protein and its RNA recognized motifs were observed., Conclusions: The result might aid for basic research and clinical application.

Objective: To identify and clone the gene encoding human M96 gene and study its expression spectrum in several blood cell lines., Methods: According to the sequence of human EST which was highly homologous to the mouse M96 gene, primers used for library screening were synthesized, then the human adult testis and fetal brain cDNA library were screened. The gene was analyzed by making use of BLAST and CLUSTAL W, and its expression spectrum was studied by multiple-cell lines Northern blot analysis. The expression change of M961 in cell differentiation was observed by use of K562 cell line induced by hemin., Results: Two cDNA clones encoding human M96 gene were isolated, identified and named as M961, and M962. They were found to be isoforms of each other. Northern, blot showed that M961 gene was expressed highly in CEM, Hel, Dami and K562 cell lines. However, during K562 cell line differentiation, process the expression of M961 elevated only slightly., Conclusions: M961 gene was expressed highly in pluripotent cell lines with erythrocytic and megakaryocytic potentials.

Human calcyclin binding protein (hCacyBP) gene was obtained by the screening of a human cDNA library. The full coding region of CacyBP was cloned into E.coli strain pET28, and then was expressed and purified through affinity chromatography. Rabbit anti-human CacyBP polyclonal antibody was obtained by immunizing rabbit with the purified human CacyBP. Western blots showed that it was expressed extensively in many tissues of mouse. The results of immunohistochemistrial staining showed that the location of CacyBP in BT325 cell line before and after differentiation changed from cytoplasm into nucleus and perinucleus cytoplasm.

Cellular RNAs in eukaryotes undergo extensive posttranscriptional modifications, but as yet only a few RNA-modifying enzymes have been identified and characterized. Here we report on the cloning of FTSJ2, a novel human gene encoding a putative RNA methyltransferase. FTSJ2 shares significant sequence homology with FtsJ/RrmJ, a recently identified Escherichia coli 23S rRNA uridine-2'-O-methyltransferase. FTSJ2 belongs to a new family of evolutionarily conserved S-adenosylmethionine-binding proteins. The gene FTSJ2 is located on chromosome 7p22 between MAD1L1 and NUDT1. It is 8 kb in length, spanning three exons. Northern blot analysis revealed that the FTSJ2 transcripts are abundant in skeletal muscle, placenta, and heart, as well as in cancer cells. Immunofluorescence staining demonstrated that FTSJ2 protein localizes to the nucleolus. Our results suggest that FTSJ2 is likely a nucleolar RNA methyltransferase involved in eukaryotic RNA processing and modification.

Host cell factor 1 (HCF-1) is a cellular transcriptional coactivator which coordinates the assembly of enhancer complex through direct interactions with viral and cellular trans-activators such as VP16, Oct-1, LZIP, and GA-binding protein. These interactions are mediated by the beta-propeller domain comprising the first 380 residues of HCF-1 with six kelch repeats. Here we describe the identification and characterization of a novel HCF-like kelch repeat protein, designated HCLP-1. HCLP-1 is a ubiquitously expressed nuclear protein which is composed almost entirely of a six-bladed beta-propeller. HCLP-1 selectively interacts with LZIP but not with VP16. The physical interaction between HCLP-1 and LZIP leads to the repression of the LZIP-dependent transcription. The HCLP-1-binding domain of LZIP maps to residues 109-315, which contain the bZIP DNA-binding motif. Electrophoretic mobility shift assay demonstrates that HCLP-1 indeed interferes with the binding of LZIP to its DNA target. Thus, HCLP-1 serves a transcriptional co-repressor function mediated through its inhibitory interaction with the LZIP transcription factor. Our findings suggest a new mechanism for transcriptional regulation by HCF-like proteins.

Purpose: To report an experimental study investigating the ability of nonporous polytetrafluoroethylene (PTFE) covering on a metallic stent to retard the development of neointimal hyperplasia (NIH)., Methods: Three groups of Hanford miniature swine underwent standardized balloon injury to both external iliac arteries. Group I animals (control) received balloon injuries only. Group II had the site of balloon injury supported by a properly sized, balloon-expandable Palmaz stent placed directly over the injury site. Group III animals received a Palmaz stent covered with PTFE graft. All animals underwent arteriography immediately after intervention and again prior to sacrifice and specimen harvest at 4 weeks. The specimens were examined grossly and histologically at the proximal, middle, and distal segments for NIH development., Results: Uncovered stents developed significantly more NIH (p < 0.0001) and greater luminal narrowing (p < 0.001) than the controls. PTFE-covered stents (group III) exhibited less NIH (p < 0.001) and luminal reduction (p < 0.01) than bare stents (group II) at the middle portion of the stent-graft, but the PTFE cover had no effect on NIH and lumen reduction at the proximal or distal ends of the prosthesis., Conclusions: PTFE-covered stents retarded NIH at 4 weeks, but only at the midportion of the devices; the covering did not prevent neointimal pannus ingrowth at the proximal and distal ends.