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The mechanism of Li-O 2 batteries is based on the reactions of lithium ions and oxygen, which hold a theoretical higher energy density of approximately 3500 W h kg -1 . In order to improve the practical specific capacity and cycling performance of Li-O 2 batteries, a catalytically active mechanically robust air cathode is required. In this work, we synthesized a freestanding catalytic cathode with RuO 2 decorated 3D web Co 3 O 4 nanowires on nickel foam. When the specific capacity was limited at 500 mA h g -1 , the RuO 2 -Co 3 O 4 /NiF had a stable cycling life of up to 122 times. The outstanding performance can be primarily attributed to the robust freestanding Co 3 O 4 nanowires with RuO 2 loading. The unique 3D web nanowire structure provides a large surface for Li 2 O 2 growth and RuO 2 nanoparticle loading, and the RuO 2 nanoparticles help to promote the round trip deposition and decomposition of Li 2 O 2 , therefore enhancing the cycling behavior. This result indicates the superiority of RuO 2 -Co 3 O 4 /NiF as a freestanding highly efficient catalytic cathode for Li-O 2 batteries., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Aim: To evaluate microstructural visual pathway damage in patients with primary glaucoma (PG) by using 3 T diffusion kurtosis imaging (DKI)., Materials and Methods: The study was approved by the ethics committee, and all participants provided written informed consent. Ten patients with PG were examined. Twenty healthy individuals served as control subjects. DKI was performed with a GE Silent 3 T magnetic resonance imaging (MRI) unit. Mean diffusivity (MD), fractional anisotropy (FA), and mean kurtosis (MK) maps were automatically created. Mean MK, MD, and FA values were calculated for each part of the visual pathway., Results: No abnormalities in the shape and signal intensity were observed along the entire visual pathway in patients and the control group on the conventional MRI. Higher MD, and lower MK and FA were observed in the optic nerves (ON), lateral geniculate nucleus (LGN), optic radiations (OR), and visual cortex (VCx) of PG patients, as compared with control subjects. A significantly higher MD was observed in the ON (p<0.01), and significantly lower FA was observed in OR (p<0.05). Additionally, significantly lower MK was observed in the ON, LGN, and VCx, except for OR (p<0.01). Changes of DKI parameters in the ON were the most distinct., Conclusion: Glaucoma is a complex neurological disease that affects the entire visual pathway. MK derived from DKI would be a better biomarkers than FA and MD in detecting microstructural damage., (Copyright © 2018 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.)

A series of dicarboxylates bridged titanium-oxo clusters with {Ti 3 (μ 3 -O)} building units have been synthesized through facile one-step solvothermal reactions. It is interesting to find that the geometric characteristics of the obtained supramolecular structures highly depend on the configuration of the applied dicarboxylate ligands. A linear dimeric [Ti 3 O] 2 L 2 complex can be constructed using flexible cyclohexanedicarboxylic acid, while the introduction of rigid 2-nitro-1,4-benzenedicarboxylic acid gives rise to a triangular [Ti 3 O] 3 L 3 structure. Moreover, unusual [Ti 3 O] 4 L 4 rectangles have been achieved with more symmetric 5-nitro-1,3-benzenedicarboxylic acid or terephthalic acid. Furthermore, a photochromic effect is observed on the obtained complexes upon UV-vis light irradiation in the presence of alcohol.

NOD2 (nucleotide-binding oligomerization domain-containing protein 2) is an internal pattern recognition receptor that recognizes bacterial peptidoglycan and stimulates host immune responses. Dysfunction of NOD2 pathway has been associated with a number of autoinflammatory disorders. To date, direct inhibitors of NOD2 have not been described due to technical challenges of targeting the oligomeric protein complex. Receptor interacting protein kinase 2 (RIPK2) is an intracellular serine/threonine/tyrosine kinase, a key signaling partner, and an obligate kinase for NOD2. As such, RIPK2 represents an attractive target to probe the pathological roles of NOD2 pathway. To search for selective RIPK2 inhibitors, we employed virtual library screening (VLS) and structure based design that eventually led to a potent and selective RIPK2 inhibitor 8 with excellent oral bioavailability, which was used to evaluate the effects of inhibition of RIPK2 in various in vitro assays and ex vivo and in vivo pharmacodynamic models.

A new naphthalenepropanoic acid analog (1) was isolated from the broth of the actinomycetes Micromonospora sp. HS-HM-036. The structure of compound 1 was determined based on MS and extensive NMR analysis. A preliminary investigation of the biological activity of compound 1 was also described.

Bismuth selenide (Bi 2 Se 3 ), with a wide bulk band gap and single massless Dirac cone at the surface, is a promising three-dimensional topological insulator. Bi 2 Se 3 possesses gapless surface states and an insulator-like bulk band gap as a new type of quantum matter. Different Bi 2 Se 3 nanostructures were prepared using electron beam evaporation with high production efficiency. Structural investigations by energy-dispersive X-ray analysis, scanning electron microscopy, and X-ray diffraction revealed the sample stoichiometries and the structural transition mechanism from nanocrystals to nanoflakes. The optical properties systematically probed and analyzed by spectroscopic ellipsometry showed strong dependence on the nanostructures and were also predicted to have structure-modifiable technological prospects. The optical parameters, plasma frequencies, scattering rates of the free electrons, and optical band gaps were related to the topological properties of the Bi 2 Se 3 nanostructures via light-matter interactions, offering new opportunities and approaches for studies on topological insulators and spintronics. The high-quality Bi 2 Se 3 nanostructures provide advantages in exploring novel physics and exploiting prospective applications.

Four titanium-oxo-clusters with cyclohex-3-ene-1-carboxylate (Cec) ligands, Ti 6 (μ 2 -O) 2 (μ 3 -O) 2 (Cec) 4 (O i Pr) 12 (PTC-91; HO i Pr = isopropanol), Ti 6 (μ 3 -O) 6 (Cec) 6 (O i Pr) 6 (PTC-92 and PTC-93) and H 2 [Ti 18 (μ 2 -O) 11 (μ 3 -O) 14 (Cec) 20 (O i Pr) 4 ] (PTC-94), have been solvothermally synthesized under different conditions, respectively. These clusters with the same Cec ligand show totally different structures, and they are stable in water and other common solvents. The Ti 6 clusters with different conformations are presented in PTC-91, PTC-92 and PTC-93, while PTC-94 is an unusual Ti 18 cluster. The photocatalytic studies reveal that PTC-91 and PTC-94 have good H 2 evolution ability.

To analyze the clinical drug use characteristics and medication rules of cervus and cucumis polypeptide injection (CCP injection) for the patients with lumbar vertebra diseases. Based on the electronic medical data of 6 990 patients with CCP injection for lumbar vertebra diseases from 25 first-class hospitals over the whole country from 2004, statistical analysis was conducted for their general information, diagnostic information, orders information, laboratory test information and treatment results. The results showed that the age distribution was highest in patients aged 45 to 64 (3 883, 55.55%); protrusion of lumbar intervertebral disc was most common among lumbar vertebra diseases (5 519, 82.50%); and TCM syndrome mainly included hemostasis. The daily dose was most between 8-12 mL, with a usual treatment course of 1-3 d. Commonly used western medicines in combined drug use included hypertonic dehydrating agent, antibiotics and nutrition nerve medicines, and the commonly used traditional Chinese medicines in combined drug use included the drugs with promoting blood circulation to remove blood stasis, kidney-tonifying and bone-building agents, and formula for clearing heat and removing toxicity, etc. The most commonly used western medicines are methylamine, adenosine, lidocaine injection, mannitol, et al, and the most commonly used Chinese medicines included Jintiange capsules, Qianggu capsules, et al. It is suggested that CCP injection for lumbar vertebrae diseases is mainly used in middle-aged and elderly patients, and is frequently used in combination with kidney-tonifying and bone-building agents, drugs with promoting blood circulation to remove blood stasis, analgesic agents and antibiotics., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

An efficient solvent-free protocol for the Buchwald-Hartwig cross-coupling reaction of aryl and heteroaryl chlorides with primary and secondary amines using the Pd(dba) 2 /ligand 1 catalytic system has been developed. Notably, the catalytic system also efficiently catalyzed the reaction under aqueous conditions.

An easily available Pd(OAc) 2 /(2-(anthracen-9-yl)-1H-inden-3-yl) dicyclohexylphosphine/toluene/ i PrOH/water catalytic system was developed, which shows high catalytic activity in the Suzuki-Miyaura cross-coupling reactions of a diverse array of aryl and heteroaryl chlorides with Pd loadings down to 0.01 mol%.

The study on microbial community composition in rhizosphere soils surrounding different order roots is of great significance for understanding the interactions between roots and microbes. Using Illumina Miseq sequencing technology, this study analyzed the differences of fungal community structure in bulk soils and rhizosphere soils surrounding different root orders of poplar (Populus × euramericana 'Neva') tree. The microbial species annotation showed that 128, 124, 130 and 101 fungal genera were classified in the rhizosphere soils around 1-2 order roots (R1), 3 order roots (R2), 4-5 order roots (R3) and in the bulk soils (NR), respectively. The differences of present fungal genera indicated a selectivity mechanism driving fungal community assembly in poplar rhizosphere soils. There were seven fungal genera with more than 1% of relative abundance in rhizosphere soils. Trichoderma was the dominant fungal genus in R1. Trichosporon and Aspergillus were the dominant fungal genera in R2 and R3, respectively. Alpha (α) diversity indices showed that the fungal diversity was significantly different among root orders. Specifically, the diversity of soil fungal community in the rhizosphere soils around lower order roots was significantly higher than that of higher order roots (P＜0.05). Beta (β) diversity indices showed that the dissimilarity of fungal community composition increased along with the root orders. All these results implied the different composition and structure of fungal community are closely related with the function of fine root orders.

Pak choi is a highly nutritious vegetable that is widely grown in China, Southeast Asia, and other parts of the world. Because it reproduces by seed, it is very important to understand the mechanism of floral organ development. Therefore, using the Chinese cabbage genome as a reference, this study analyzed the expression profiles of shoot apex genes at flower bud differentiation stages 1 and 5, in order to identify genes related to floral organ development. The results showed that the proportion of mapped genes was high, with 84.25 and 83.80% of clean reads from the two sample saligned to the reference genome, respectively. A total of 525 differentially expressed genes (DEGs) were identified, 224 of which were upregulated and 301 were downregulated. The expression levels of genes homologous to Chinese cabbage flowering genes were also analyzed at stages 1 and 5; the expression levels of Bra012997 (ap1), Bra000393 (SOC1), and Bra004928 (SOC1) were significantly upregulated at stage 5, suggesting that these three genes positively regulate floral development in pak choi. DEGs involved in floral organ development were analyzed with homologous genes from Arabidopsis thaliana; the homologous genes Bra029281 (AGL42), Bra026577 (ARPN), Bra022954 (SPL3), Bra029293 (ARF2), Bra007978 (AtRLP12), Bra033221 (SPL8), Bra008037 (LOX4), Bra001598 (IAA19), Bra003892 (PATL1), Bra038778 (AT4G21323), Bra025315 (KLCR2), and Bra013906 (DTX35) are directly related to floral organ development in Arabidopsis, suggesting that these genes have corresponding functions during flower organ development in pak choi, and could be candidates for further genetic research. These results provide a foundation for research on the molecular mechanism of flower organ development in pak choi and other Brassica rapa vegetables.

MYBA2 transcription factor (Myb-related gene) affects the coloring in grapevine berry and plays an important role in the biosynthesis of anthocyanin. The MYBA2 gene was cloned from Vitis vinifera L. cv. Cabernet Sauvignon and polyclonal antibodies for VvmybA2 were prepared. The VvmybA2 gene expression patterns were observed in seven tissues (the leaf, stem, flower, bud, root, berry, and tendril) and during the berry development stage at transcriptional and translational levels, respectively. The results indicated that the expression of VvmybA2 was approximately 11-fold higher in the berry than that in the other six tissues, and increased rapidly from 60 days after full bloom reaching a maximum on day 80. Furthermore, both the anthocyanin content and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT) gene expression levels increased rapidly 60 days after full bloom. Moreover, correlation analysis indicated that the transcriptional and translational expression levels of the VvmybA2 gene were significantly positively correlated with not only UFGT and DFR genes but also with the anthocyanin content during berry development. These results suggested that VvmybA2 could not only regulate the transcription of both UFGT and DFR but also is involved in the expression of the UFGT gene associated with color determination in grape berries.

In situ external magnetic field was applied during the synthesis of FePt nanoparticles via a chemical solution method. FePt nanoparticle films were prepared on Si by a drop-coating method with and without a magnetic field. Annealing at 700 °C in reductive atmosphere was explored to obtain ferromagnetic FePt L10 phase. The effect of in situ-applied magnetic field on the structure, morphology, and magnetic properties of FePt nanoparticle films was characterized. It is found that the applied magnetic field during the chemical synthesis of FePt nanoparticles plays a key role in the crystallinity and magnetic property of FePt nanoparticle films. As-synthesized FePt nanoparticles under the magnetic field are monodispersed and can be self-assembled over a larger area by a dropping method. The applied magnetic field during the synthesis of FePt nanoparticles not only significantly improves the nanoparticles' c-axis preferred orientation but also benefits the phase transition of FePt nanoparticles from face-centered cubic to face-centered tetragonal structure during the annealing process. The FePt nanoparticle films derived under magnetic field also show some magnetic anisotropy.

To investigate factors influencing the intranasal absorption of rivastigmine hydrogen tartrate (RHT), we studied the pharmacokinetics of RHT after intranasal administration and evaluated its brain targeting behavior. In situ rat nasal perfusion model was used in the study and pH impact was examined on the intranasal absorption of RHT. High performance liquid chromatography (HPLC) method was established to measure RHT concentration in the plasma and brain tissue after intranasal and intravenous administration. The pharmacokinetic parameters, drug targeting index(DTI), and nose-to-brain direct transport percentage (DTP) were calculated. It was demonstrated that the intranasal absorption mechanism of RHT was passive diffusion. The absorption rate was highest at pH 6.0. The absolute bioavailability of intranasally administrated RHT was 73.58%. Compared with that of intravenous administration, RHT absorption into the brain was faster and more efficient after intranasal delivery, and the DTI value was 195.27% of intravenous injection. Moreover, 48.79% of the drug can be absorbed directly from the nose into the brain without systematic circulation. Meanwhile, drug elimination half-time in the brain was prolonged by 1.4 fold compared to that of intravenous injection. In conclusion, intranasal administration of RHT not only improves drug absorption into the system, but also enhances drug absorption rate and content in the brain remarkably, which is an advantage in the treatment of central nervous system-related diseases.

Chagas disease, leishmaniasis and sleeping sickness affect 20 million people worldwide and lead to more than 50,000 deaths annually. The diseases are caused by infection with the kinetoplastid parasites Trypanosoma cruzi, Leishmania spp. and Trypanosoma brucei spp., respectively. These parasites have similar biology and genomic sequence, suggesting that all three diseases could be cured with drugs that modulate the activity of a conserved parasite target. However, no such molecular targets or broad spectrum drugs have been identified to date. Here we describe a selective inhibitor of the kinetoplastid proteasome (GNF6702) with unprecedented in vivo efficacy, which cleared parasites from mice in all three models of infection. GNF6702 inhibits the kinetoplastid proteasome through a non-competitive mechanism, does not inhibit the mammalian proteasome or growth of mammalian cells, and is well-tolerated in mice. Our data provide genetic and chemical validation of the parasite proteasome as a promising therapeutic target for treatment of kinetoplastid infections, and underscore the possibility of developing a single class of drugs for these neglected diseases., Competing Interests: Patents related to this work has been filed (WO 2015/095477 A1, WO 2014/151784 A1, WO 2014/151729). Several authors own shares of Novartis.

Through the labile coordination sites of a robust phosphonate-stabilized titanium-oxo cluster, 14 O-donor ligands have been successfully introduced without changing the cluster core. The increasing electron-withdrawing effect of the organic species allows the gradual reduction of the bandgaps of the {Ti6 } complexes. Transition-metal ions are then incorporated by the use of bifunctional O/N-donor ligands, organizing these {Ti6 } clusters into polymeric structures. The coordination environments of the applied metal ions show significant influence on their visible-light adsorption. Both the above structural functionalizations also tune the photocatalytic H2 production activities of these clusters. This work provides a systematic bandgap engineering study of titanium-oxo clusters, which is important not only for their future photocatalytic applications, also for the better understanding of the structure-property relationships., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

We present the formation of the largest titanium-oxo cluster, [Ti42(μ3-O)60(OiPr)42(OH)12)](6-), with the first fullerene-like Ti-O shell structure. The {Ti42O60} core of this compound exemplifies the same icosahedral (Ih) symmetry as C60, the highest possible symmetry for molecules. According to the coordination environments, the Ti centers in this cluster can be arranged into a Platonic {Ti12} icosahedron and an Archimedean {Ti30} icosidodecahedron. The solution stability of this cluster was confirmed by electrospray ionization mass spectrometry. The spherical body of the {Ti42O60} core has an inside diameter of 1.05 nm and an outside diameter of 1.53 nm, which could be directly visualized by high-resolution transmission electron microscopy. Our results demonstrate that titanium oxide can also form fullerene-like shell structures.

The objectives of this study are to prepare resveratrol loaded mixed micelles composed of poloxamer 403 and poloxamer 407, and optimize the formulation in order to achieve higher drug solubility and sustained drug release. Firstly, a thin-film hydration method was utilized to prepare the micelles. By using drug-loading, encapsulation yield and particle size of the micelles as criteria, influence of three variables, namely poloxamer 407 mass fraction, amount of water and feeding of resveratrol, on the quality of the micelles was optimized with a central composite design method. Steady fluorescence measurement was carried out to evaluate the critical micelle concentration of the carriers. Micelle stability upon dilution with simulated gastric fluid and simulated intestinal fluid was investigated. The in vitro release of resveratrol from the mixed micelles was monitored by dialysis method. It was observed that the particle size of the optimized micelle formulation was 24 nm, with drug-loading 11.78%, and encapsulation yield 82.51%. The mixed micelles increased the solubility of resveratrol for about 197 times. Moreover, the mixed micelles had a low critical micelle concentration of 0.05 mg · mL(-1) in water and no apparent changes in particle size and drug content were observed upon micelles dilution, indicating improved kinetic stability. Resveratrol was released from the micelles in a controlled manner for over 20 h, and the release process can be well described by Higuchi equation. Therefore, resveratrol-loaded poloxamer 403/407 mixed micelles could improve the solubility of resveratrol significantly and sustained drug release behavior can be achieved.

To examine the effect of postharvest ultraviolet C (UV-C) irradiation on flavanol polyphenol accumulation in the grape berry, we investigated total flavanol polyphenol content, the enzyme activity of leucoanthocyanidin reductase (LAR), and transcription of Vv lar1 and Vv lar2 using spectrophotometry, real-time polymerase chain reaction, and western blot analysis in 5-year-old Vitis vinifera L. cv. Cabernet Sauvignon plants. Our results indicated that the accumulation of flavanol polyphenol reached its highest value when exposed to UV-C irradiation for 30 min. Additionally, UV-C irradiation induced the transcription of Vv lar1 and Vv lar2 and the synthesis of LAR1 and LAR2 proteins, resulting in increased accumulation of flavanol polyphenol in the grape berry. Moreover, these effects were associated with the length of time of UV-C irradiation.

Objective: To examine factors that may have impact on the Mini-Mental State Examination (MMSE) screening validity, which could lead to further establishing the general model of the MMSE score in Chinese health elderly and to improve the screening accuracy of the existing MMSE reference., Methods: Based on the data of the Chinese Longitudinal Healthy Longevity Survey (CLHLS), the MMSE scores of 19,117 normal elderly and 137 dementia patients who met the inclusion criteria were used for the analysis. The area under the curve (AUC) and validity indexes were used to compare the screening accuracy of various criteria. Multiple linear regression was used to identify factors that had impact on the MMSE score for both the normal and dementia elderly. Descriptive analysis was performed for differences in the MMSE scores by age trends and gender between the normal and dementia elderly., Results: The AUC of MMSE was ≥0.75(P<0.05). The MMSE score of the normal elderly declined nonlinearly as the age grew older(male: R2=0.924, P<0.05; female: R2=0.951, P<0.05), and increased nonlinearly as the education level rose(male: R2=0.948, P<0.05; female: R2=0.859, P< 0.05). The females had significantly lower MMSE scores than the males, with a faster decline trend with age than the males (95%CI of female partial regression coefficient was not overlapped with 95%CI of male partial regression coefficient). The dementia elderly showed a much lower MMSE score (male: difference of Z score:-1.573, P<0.05; female: difference of Z score:-1.222, P<0.05) and tended to with a faster decline speed than that of the normal elderly (95%CI of dementia partial regression coefficient included 95%CI of normal partial regression coefficient)., Conclusion: The screening validity of MMSE in CLHLS is not affected by educational level. The analysis of factors that may impact on the MMSE screening validity are gender, age, vision and residence which with validity identification. These four factors can be used as assist tool of MMSE in the screening of dementia to improve the screening accuracy.

Background: Considering the key role of TF in coagulation of sepsis or acute lung injury (ALI), we investigated whether berberine (BBR) could inhibit TF expression and procoagulant activity and explored its possible mechanism., Methods: The effects of berberine on the expression, procoagulant activity of TF and related signal pathways induced by lipopolysaccharide (LPS) were observed in THP-1 cells., Results: Our results showed that berberine could inhibit LPS-induced TF activity and expression, and down-regulate NF-κB, Akt and MAPK/JNK/p38/ERK pathways., Conclusion: Berberine inhibits TF expression and related pathway, which provides some new insights on its mechanism for sepsis treatment., (Copyright © 2014 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.)

Objective: To investigate the effects of alkaloid monomers from Gelsemium elegans on proliferation of HepG2 cell in vitro and the possible mechanism., Methods: MTT assay was used to measure the inhibitory of three alkaloid monomers on HepG2 cell in vitro. The most effective fraction was chosen to test whether the effect was in time-and dose-dependent manner. The morphological changes were observed by the light microscope and the cell cycle alteration through the flow cytometric assay. The activity of Caspase-3, Caspase-8 and Caspase-9 were detected by a Caspases colorimetric assay kit., Results: The results showed that koumine, Gelsemine and Gelsenicine could significantly inhibit the proliferation of HepG2 cell and Gelsenicine, the most effective fraction, was clearly in dose- and time-dependent manners, while exhibited low cytotoxicity to the Vero cell. The cell treated with Gelsenicine for 48 h showed distinctive morphological changes. The cells treated with 200 and 400 microg/mL shrinked and fell off from the bottom. At the same time, the cells were arrested at S phase and the apoptosis increased apparently. The activity of Caspase-3, Caspase-8 and Caspase-9 was increased in a dose-dependent manner., Conclusion: Three alkaloid monomers from Gelsemium elegans, especially Gelsenicine, could inhibit proliferation of HepG2 cell obviously. The mechanism may be related to cell cycle arrest and activation of Caspase-3, Caspase 8 and Caspase-9.

A novel δ-endotoxin gene was cloned from a Bacillus thuringiensis strain with activity against Locusta migratoria manilensis by PCR-based genome walking. The sequence of the cry gene was 3,432 bp long, and it encoded a Cry protein of 1,144 amino acid residues with a molecular mass of 129,196.5 kDa, which exhibited 62% homology with Cry7Ba1 in the amino acid sequence. The δ-endotoxin with five conserved sequence blocks in the amino-terminal region was designated Cry7Ca1 (GenBank accession no. EF486523). Protein structure analysis suggested that the activated toxin of Cry7Ca1 has three domains: 227 residues forming 7 α-helices (domain I); 213 residues forming three antiparallel β-sheets (domain II); and 134 residues forming a β-sandwich (domain III). The three domains, respectively, exhibited 47, 44, and 34% sequence identity with corresponding domains of known Cry toxins. SDS-PAGE and Western blot analysis showed that Cry7Ca1, encoded by the full-length open reading frame of the cry gene, the activated toxin 1, which included three domains but without the N-terminal 54 amino acid residues and the C terminus, and the activated toxin 2, which included three domains and N-terminal 54 amino acid residues but without the C terminus, could be expressed in Escherichia coli. Bioassay results indicated that the expressed proteins of Cry7Ca1 and the activated toxins (toxins 1 and 2) showed significant activity against 2nd instar locusts, and after 7 days of infection, the estimated 50% lethal concentrations (LC₅₀s) were 8.98 μg/ml for the expressed Cry7Ca1, 0.87 μg/ml for the activated toxin 1, and 4.43 μg/ml for the activated toxin 2. The δ-endotoxin also induced histopathological changes in midgut epithelial cells of adult L. migratoria manilensis.

Objective: To understand the risk behavioral networks of newly reported HIV infections in Taizhou prefecture, Zhejiang province., Methods: Newly reported HIV infections from May 2008 through March 2010 in Taizhou prefecture were invited to participate in a cross-sectional survey which requested numbers and contact information of individuals with whom they had had high risk contacts as well as risk behavioral acts with them. Persons having had risky contacts with HIV-infected cases were thereof approached to participate in this survey and to receive HIV testing. Those who tested positive for HIV were subject to further round of the surveys., Results: A total of 267 HIV cases were newly reported during this study period. Among them, 191 participated in the survey and served as 'index cases', including 158 cases identified from routine HIV surveillance programs and 33 cases identified from the present survey. Heterosexual transmission was the primary transmission route (74.9%, or 143/191), followed by homosexual transmission (19.4%, or 37/191) and injection drug use (5.8%, or 11/191). These 191 HIV cases reported a total of 1152 individuals with whom they had had risky contacts. They were able to provide contact information of 461 risk contacts. Of them, 129 received HIV testing and 61 (47.3%) tested positive for HIV. HIV prevalence was the highest among spouses or long-term sex partners of HIV cases (45.6%, or 47/103) and male sex partners of HIV-infected men having sex with men (MSM) (60.0%, or 12/20). Condom use was very low among them, with only 33.9% consistently using condoms for sex., Conclusion: Newly reported HIV infections in Taizhou prefecture reported a large and complicated risk behavioral networks and low condom use, suggesting a potential risk of HIV among these connected people, especially among MSM. Much efforts are needed to intervene these high risk subgroups and high risk behavioral networks.

ALK (anaplastic lymphoma kinase) is an RTK (receptor tyrosine kinase) of the IRK (insulin receptor kinase) superfamily, which share an YXXXYY autophosphorylation motif within their A-loops (activation loops). A common activation and regulatory mechanism is believed to exist for members of this superfamily typified by IRK and IGF1RK (insulin-like growth factor receptor kinase-1). Chromosomal translocations involving ALK were first identified in anaplastic large-cell lymphoma, a subtype of non-Hodgkin's lymphoma, where aberrant fusion of the ALK kinase domain with the NPM (nucleophosmin) dimerization domain results in autophosphosphorylation and ligand-independent activation. Activating mutations within the full-length ALK kinase domain, most commonly R1275Q and F1174L, which play a major role in neuroblastoma, were recently identified. To provide a structural framework for understanding these mutations and to guide structure-assisted drug discovery efforts, the X-ray crystal structure of the unphosphorylated ALK catalytic domain was determined in the apo, ADP- and staurosporine-bound forms. The structures reveal a partially inactive protein kinase conformation distinct from, and lacking, many of the negative regulatory features observed in inactive IGF1RK/IRK structures in their unphosphorylated forms. The A-loop adopts an inhibitory pose where a short proximal A-loop helix (alphaAL) packs against the alphaC helix and a novel N-terminal beta-turn motif, whereas the distal portion obstructs part of the predicted peptide-binding region. The structure helps explain the reported unique peptide substrate specificity and the importance of phosphorylation of the first A-loop Tyr1278 for kinase activity and NPM-ALK transforming potential. A single amino acid difference in the ALK substrate peptide binding P-1 site (where the P-site is the phosphoacceptor site) was identified that, in conjunction with A-loop sequence variation including the RAS (Arg-Ala-Ser)-motif, rationalizes the difference in the A-loop tyrosine autophosphorylation preference between ALK and IGF1RK/IRK. Enzymatic analysis of recombinant R1275Q and F1174L ALK mutant catalytic domains confirms the enhanced activity and transforming potential of these mutants. The transforming ability of the full-length ALK mutants in soft agar colony growth assays corroborates these findings. The availability of a three-dimensional structure for ALK will facilitate future structure-function and rational drug design efforts targeting this receptor tyrosine kinase.

To study the effects of major components of Maijunan tablets, puerarin (Pue) and rhynchophylline (Rhy) on the transport of hydrochlorothiazide (Hct) Caco-2 cell monolayer model, the transport parameters of Hct, such as apparent permeability coefficient (P(app) (B --> A) and P(app) (A --> B)) and the ratio of P(app) (B --> A) versus P(app) (A --> B), were studied and compared when Hct was used solely and co-used with Pue and/or Rhy. The effects of drug concentrations, conveying times, P-glyprotein (P-gp) inhibitor verapamil and conveying Liq pH values on the transport of Hct in the above conditions were also investigated. The results indicated that the absorption of Hct in Caco-2 cell monolayer model could be a carrier-mediated active transport, along with the excretion action mediated by P-gp. Pue can decrease the excretion action of Hct mediated by P-gp, and Rhy had no significant effect on the transport of Hct. The co-use of Hct, Pue and Rhy enhanced the absorption of Hct. Meanwhile, conveying Liq pH value had significant influence on the transport of Hct. The absorption of Hct at pH 6.0 was higher than that at pH 7.4.

Objective: To explore the sero-prevalence and risk factors for herpes simplex virus 2 (HSV-2) infection and unprotected sexual behavior in an ethnically diverse population of HIV-infected subjects in a county of Yunnan province., Methods: HIV-infected individuals attending for routine follow-up by local Center for Disease Control and Prevention (CDC) were recruited to participate in the study under 'informed consent'. A face-to-face questionnaire interview was administered to each participant. Blood was drawn for HSV-2 testing by HerpeSelect HSV-2 ELISA (Focus Diagnostics) and CD(4)(+) T counting., Results: A total of 300 HIV-infected individuals participated in the study. The mean age of the subjects was 37.6 years with 76.7% as males. Ethnically, Han, Dai and Jingpo accounted for 44.3%, 37.3% and 16.0% of the sample, respectively. Half of the subjects reported HIV acquisition through injection drug use. The sero-prevalence of HSV-2 was 35.0%. Results from multiple logistic regression analysis indicated that individuals who acquired HIV through heterosexual contact were more likely to be HSV-2 positive than those who acquired HIV through injection drug use (OR = 4.244, 95%CI: 1.924 - 9.364), whereas Dai (OR = 0.300, 95%CI: 0.152 - 0.593) and Jingpo (OR = 0.376, 95%CI: 0.167 - 0.850) were less likely to be HSV-2 positive than the Hans. Among 105 people who were co-infected with HIV/HSV-2, 60 had sexual intercourses in the past 3 months and 41.7% of them reported no or inconsistent use of condoms. Most unprotected sexual contacts occurred within married couples., Conclusion: HSV-2 infection was highly prevalent among HIV-infected individuals in this county, and a significant proportion of HIV/HSV-2 co-infected subjects engaged in unprotected sex. HSV-2 testing, behavioral and biomedical interventions among HIV-infected individuals and their sexual partners should be involved in the local HIV prevention and control programs.

Objective: To assess the adherence, immunologic and survival responses in HIV-infected patients receiving free antiretroviral therapy (ART)., Methods: All adult HIV-infected patients in Wenxi county who started antiretroviral treatment (ART) between 01 July 2001 and 31 December 2006 and aged above 18 years were included in this study. Epidemiological survey and laboratory tests were performed before, 0.5 months after, 1 months after, 2 months after and every 3 months after initiation of ART to recognize the adherence, efficacy (CD(4)(+) T cell counts) and survival to the regimens., Results: The median follow-up time period was 16.5 months (Interquartile: 15.5 - 20.8 months). At baseline, the median of CD(4)(+) T cell counts were 154 cells/microl (Interquartile: 81 - 212 cells/microl). Treatment was effective in most of the patients, the CD(4)(+) T cell count of patients increased after the initiation of ART. The maximum increase was recorded at month 3, from the median of 154 cells/microl to 220 cells/microl (P < 0.001), and thereafter the count remained stable. When comparing with patients with baseline CD(4)(+) T cell count > or = 100 cells/microl, those with baseline CD(4)(+) T cell count < 100 cells/microl showed a higher mean increase in the first three months of treatment. The cumulative probability rates of remaining alive were 0.94, 0.88 and 0.87 at 3, 12, 24 months, respectively. In multivariate Cox's proportional hazard models, after adjustment for the type ofinitial regimens (NVP vs. EFV/IDV), CD(+)(4) T cell count of less than 50 cells/microl (vs. 50 cells/microl or more) was strongly associated with death hazard ratio 0.21 (95%CI: 0.06 - 0.68)., Conclusion: Our data showed that ART was effective for improving immunologic response of adult patients with HIV/AIDS. CD(4)(+) T cell count at initiation was associated with survival time in patients starting ART, suggesting that monitoring of CD(4)(+) T count should be strengthened to early initiate antiretroviral therapy for HIV-infected patients.

Objective: To study the prevalence of human immunodeficiency virus (HIV), sexually transmitted infection (STI), risk behavior and the sexual networks among men who have sex with men (MSM) in Taizhou city, Zhejiang province., Methods: A cross-sectional study was applied with venue-based sampling in 2 MSM gathering sites in Taizhou. 'Informed Consent' principle was applied and MSM were studied through a structured questionnaire. Blood samples were collected from those who accepted free and confidential HIV/STI counseling and then tested for HIV, syphilis, HCV and HSV-2 antibodies with ELISA. HIV positive sera were certified with western blot., Results: 106 MSM were investigated and 97 qualified questionnaires were collected. 25.0% (23/92) of these MSM have ever had 1 female sex partner while 47.8% (44/92) had 2 or more. 14.3% (13/91) of them reported having had 1 male partner who had engaged in anal sex and 80.2% (73/91) had 2 or more. 22.1% (19/86) of them had participated in group sex but 62.5% (55/88) of them did not always use condom when having anal intercourse. 15.1% (14/93) of them had 1 oral sex partner while 75.3% (70/93) having 2 or more. 38.9% (37/95) of them had sex with female sex worker, and 35.5% (33/93) had sex with male-to-male sex worker. 15.3% (13/85) of them had once been male-to-male sex worker themselves. 3.9% (3/77) of them were found HIV positive in blood tests, with 24.7% (18/73) positive of syphilis, 15.1% (11/73) positive of HSV-2 but HCV appeared to be negative. 46 cases reported their egocentric recognition networks, with mean degree of 5.91 (ranging 0 - 10), and mean density of 0.548 (ranging 0.000 - 1.000). 43 sexual networks were identified, with mean degree of 2.70 (ranging 0 - 10), and mean density of 0.246 (ranging 0.000 - 1.000)., Conclusion: Risk behaviors, such as multiple sex partners, low proportion of condom use and commercial sex engagement, both with heterosexuals and homosexuals, were extensively existed among MSM in Taizhou, and the prevalence of HIV/STI was relatively high. Their sexual networks seemed complicated but there might be in place of some substantially isolated MSM groups with high risk of HIV/STI infection. More study should be applied to identify the relationship between sexual networks and HIV/STI transmission.

Objective: To compare the event-related potentials (ERPs) waves of verbs and prepositions in the brain., Methods: We recorded ERPs in the brain while participants judged the legality of the collocation for verbs and prepositions., Results: Both verbs and prepositions elicited a negativity at the frontal site in 230-330 ms and 350-500 ms window. No difference was seen in 230-330 ms and 350-500 ms window; In difference waves, a negativity was elicited in the left and right hemisphere at about 270-400 ms and 470-600 ms window for both open and closed-class words., Conclusion: These may demonstrate that prepositions in modern Chinese are probably not a separate class from verbs and that N280 may be not a specific component for only prepositions (or closed-class words).

Treatment options for androgen-independent prostate cancer cells are limited. Therefore, it is critical to identify agents that induce death of both androgen-responsive and androgen-insensitive cells. Here we demonstrate that a product of plant cell walls, pectin, is capable of inducing apoptosis in androgen-responsive (LNCaP) and androgen-independent (LNCaP C4-2) human prostate cancer cells. Commercially available fractionated pectin powder (FPP) induced apoptosis (approximately 40-fold above non-treated cells) in both cell lines as determined by the Apoptosense assay and activation of caspase-3 and its substrate, poly(ADP-ribose) polymerase. Conversely, citrus pectin (CP) and the pH-modified CP, PectaSol, had little or no apoptotic activity. Glycosyl residue composition and linkage analyses revealed no significant differences among the pectins. Mild base treatment to remove ester linkages destroyed FPP's apoptotic activity and yielded homogalacturonan (HG) oligosaccharides. The treatment of FPP with pectinmethylesterase to remove galacturonosyl carboxymethylesters and/or with endopolygalacturonase to cleave nonmethylesterified HG caused no major reduction in apoptotic activity, implicating the requirement for a base-sensitive linkage other than the carboxymethylester. Heat treatment of CP (HTCP) led to the induction of significant levels of apoptosis comparable to FPP, suggesting a means for generating apoptotic pectic structures. These results indicate that specific structural elements within pectin are responsible for the apoptotic activity, and that this structure can be generated, or enriched for, by heat treatment of CP. These findings provide the foundation for mechanistic studies of pectin apoptotic activity and a basis for the development of pectin-based pharmaceuticals, nutraceuticals, or recommended diet changes aimed at combating prostate cancer occurrence and progression.

HIV/AIDS in China has entered a critical stage of rapid and widespread increase. It has been estimated that more than one million people in China have been infected with HIV and the rate of increase tops the world. The number could swell to 10 million by 2010 if more intense and effective preventive measures are not adopted immediately. Sex between men has been a mostly 'hidden' source of the spread of HIV in China. Homosexuality is no longer a criminal act in China, however, traditional 'official-led' so-called peer education programmes among men who have sex with men (MSM) have little effect in adopting and diffusing a key message to their networks. This is because the climate in HIV prevention through community-based advocacy among MSM has not been substantially changed which is due to these men still facing strong opposition and resistance from society, as a quite marginalized population in China. This study carried out in Chengdu is the first to explore how to use a socially and culturally appropriate participatory communication to promote safer sex behaviour with gay men and MSM in Chengdu, China. The study examined effectiveness of peer-led health message diffusion in promoting condom use through a participatory communication approach among these men in the programme. Key findings showed that the peer-based participatory communication strategy was effective for encouraging condom use with casual sexual partners in the intervention group. There was no significant change in the comparison group. It indicates that participatory involvement is the major driving force for HIV-related safer sex behaviour change and can be recommended to promote safer sex practice among gay men and MSM in their broad contexts.

Objective: To determine the correlates of and willingness to participate(WTP) in HIV voluntary counseling and testing (VCT) among migrants in Shanxi province., Methods: Through quota sampling, a total of 1280 migrants were recruited from 6 different venues where migrants were most likely employed to administer a face-to-face anonymous interview., Results: Knowledge of VCT among migrants was low. Only 56.6% of migrants had heard of HIV voluntary counseling and testing. 64.5% reported willingness to accept VCT after participants were informed that HIV counseling and testing was offered free of charge Centers for Disease Prevention and Control (48.7%), hospital (27.0%) and home (18.5%) were most preferred places for VCT service. Logistic regression analysis indicated that the willingness was associated with having a history of HIV test, having better knowledge of HIV, no discrimination against people with HIV, and high HIV/AIDS perception., Conclusion: Community-based health education programs for improving HIV/AIDS knowledge, promoting acceptance of VCT and eliminating discrimination to HIV-infected persons were called for.

Fourteen of 28 Trichomonas vaginalis isolates collected from patients in Guangzhou, China from 2003 to 2004 were found to be naturally infected with Mycoplasma hominis, as determined by PCR using specific primers. In vitro metronidazole sensitivity assay of the 28 isolates revealed four displaying low susceptibility [minimum lethal concentration (MLC)= approximately 13-25 microg/ml] and another four displaying high resistance (MLC=50-100 microg/ml). The overwhelming majority of these resistant isolates (7/8) were mycoplasma-infected. The mean of MLCs of mycoplasma-infected isolates is approximately 10-fold higher than the mean of noninfected isolates (p=0.029). Sequence analyses of PCR-amplified small subunit-large subunit rRNA interspacer regions (ITS1/5.8S/ITS2) revealed that 23 of the 28 samples are identical, the remaining five being separable into two groups, each with a single point mutation. These internal transcribed spacer sequence variants are associated neither with mycoplasma infection nor with drug resistance. In contrast, random amplified polymorphic DNA analyses of DNAs using 10 different primers showed that the drug-resistant isolates are clustered together in association with mycoplasma infection, albeit more loosely. Taken together, the results obtained from this study suggest that in vitro metronidazole resistance of T. vaginalis is related to mycoplasma infection of this protozoan.

Rhizobia are nitrogen-fixing bacteria that establish endosymbiotic associations with legumes. Nodule formation depends on various bacterial carbohydrates, including lipopolysaccharides, K-antigens, and exopolysaccharides (EPS). An acidic EPS from Rhizobium sp. strain NGR234 consists of glucosyl (Glc), galactosyl (Gal), glucuronosyl (GlcA), and 4,6-pyruvylated galactosyl (PvGal) residues with beta-1,3, beta-1,4, beta-1,6, alpha-1,3, and alpha-1,4 glycoside linkages. Here we examined the role of NGR234 genes in the synthesis of EPS. Deletions within the exoF, exoL, exoP, exoQ, and exoY genes suppressed accumulation of EPS in bacterial supernatants, a finding that was confirmed by chemical analyses. The data suggest that the repeating subunits of EPS are assembled by an ExoQ/ExoP/ExoF-dependent mechanism, which is related to the Wzy polymerization system of group 1 capsular polysaccharides in Escherichia coli. Mutation of exoK (NGROmegaexoK), which encodes a putative glycanase, resulted in the absence of low-molecular-weight forms of EPS. Analysis of the extracellular carbohydrates revealed that NGROmegaexoK is unable to accumulate exo-oligosaccharides (EOSs), which are O-acetylated nonasaccharide subunits of EPS having the formula Gal(Glc)5(GlcA)2PvGal. When used as inoculants, both the exo-deficient mutants and NGROmegaexoK were unable to form nitrogen-fixing nodules on some hosts (e.g., Albizia lebbeck and Leucaena leucocephala), but they were able to form nitrogen-fixing nodules on other hosts (e.g., Vigna unguiculata). EOSs of the parent strain were biologically active at very low levels (yield in culture supernatants, approximately 50 microg per liter). Thus, NGR234 produces symbiotically active EOSs by enzymatic degradation of EPS, using the extracellular endo-beta-1,4-glycanase encoded by exoK (glycoside hydrolase family 16). We propose that the derived EOSs (and not EPS) are bacterial components that play a crucial role in nodule formation in various legumes.

Objective: To investigate and clarify whether the genetic susceptibility to women with hypertensive disorder complicating pregnancy or pre-eclampsia is associated with polymorphisms and couple sharing rate of transporter associate with antigen processing genes(TAP)., Methods: One hundred and two severe pre-eclampsia women and their spouses served as study group, and 200 normal pregnant women and their spouses were selected as control group. All pregnant women were primipara with single fetus. Genomic DNA was extracted from 2 mL cubital venous blood. We used the amplification refractory mutation system polymerase chain reaction(ARMS-PCR) to characterize TAP gene locus 333, 637, 379, 565, 665., Results: We observed eleven TAP haplotypes. There were four kinds of haplotypes(1A-1D) existing in TAP1, and seven kinds of haplotypes(2A-2G) existing in TAP2. The gene frequencies of TAP2B(Chi2=9.19, P<0.01, RR=4.18) and TAP2F(Chi2=5.34, P<0.05, RR=4.63) of patient group with pre-eclampsia were significantly higher as compared with control group. The analyses of some TAP haplotypes such as TAP1B(Chi2=4.87, P<0.05, RR=3.14), TAP1C(Chi2=5.42, P<0.05, RR=4.90), TAP2B(Chi2=9.65, P<0.01, RR=5.39) showed that the couple sharing rate of pre-eclampsia women and their spouses had statistically a highly significant increase in comparison with that of controls., Conclusion: Our data suggest that the presence of TAP2B or TAP2F haplotypes should be considered as a risk increased to pregnant women being susceptible to hypertensive disorder complicating pregnancy; and also the elevated couple sharing rates of TAP1B, TAP1C and TAP2B genes will increase the opportunity or possibility of pregnant women suffering from pre-eclampsia disease.

Aim: To explore the possibility of repression of chloromycetin (Cm) acyl transferase by using external guided sequence (EGS) in order to converse the clinical E coli isolates from Cm- resistant to Cm- sensitive., Methods: EGS directed against chloromycetin acetyl transferase gene (cat) was cloned to vector pEGFP-C1 which contains the kanamycin (Km) resistance gene. The recombinant plasmid pEGFP-C1+EGScat1+cat2 was constructed and the blank vector without EGS fragment was used as control plasmids. By using the CaCl(2) transformation method, the recombinant plasmids were introduced into the clinically isolated Cm resistant but Km sensitive E coli strains. Transformants were screened on LB agar plates containing Km. Extraction of plasmids and PCR were applied to identify the positive clones. The growth curve of EGS transformed bacteria cultured in broth with Cm resistance was determined by using spectrophotometer at A(600). Drug sensitivity was tested in solid culture containing Cm by using KB method., Results: Transformation studies were carried out on 16 clinically isolated Cm-resistant (250 microg/mL of Cm) E coli strains by using pEGFP-C1-EGScat1cat2 recombinant plasmid. Transformants were screened on LB-agar plates containing Km after the transformation using EGS. Of the 16 tested strains, 4 strains were transformed successfully. Transformants with EGS plasmid showed growth inhibition when grown in liquid broth culture containing 200 microg/mL of Cm. In drug sensitivity test, these strains were sensitive to Cm on LB-agar plates containing 200 microg/mL of Cm. Extraction of plasmids and PCR amplification showed the existence of EGS plasmids in these four transformed strains. These results indicated that the Cat of the four clinical isolates had been suppressed and the four strains were converted to Cm sensitive ones., Conclusion: The EGS directed against Cat is able to inhibit the expression of Cat, and hence convert Cm-resistant bacteria to Cm-sensitive ones. Thus, the EGS has the capability of converting the phenotype of clinical drug-resistant isolates strains to drug-sensitive ones.

Objective: To study the genotype of swines isolated from humans and their relationships with hepatitis E virus (HEV) in the rural areas of southern China., Methods: Specimens collected from normal people with HEV-IgM positive, acute hepatitis E patients and from swine in the same area were detected for HEV RNA using RT-nPCR with ORF2 primers. The positive PCR products were cloned and sequenced., Results: 13 out of the 132 samples from swine stool, 4 of 26 HEV-IgM positive sera of normal people and 1 of 4 acute hepatitis E patients' stool sample and sera were tested positive for HEV RNA. Data from sequence analysis showed that the identity at nucleotide level was 89.3%-100.0% among the 10 isolates which shared 78.7% - 84.7%, 83.3% - 85.3%, 76.0% - 80.0% and 84.7% - 95.3% nucleotide sequence identity with HEV genotype I, II, III and IV respectively in the region (nt6317- 6466)., Conclusion: HEV circulating in humans and swine in the area belonged to genotype IV.

Objective: To explore the effective strategies for human immunodeficiency virus (HIV) surveillance and identification in rural areas of central China., Methods: In a selected rural prefecture area of central China, an epidemiological investigation was conducted for all reported HIV/AIDS cases. A historical and analytic review was performed, with particular interests in examining the HIV epidemic reporting and identification system., Results: Among all 626 reported HIV-infected individuals in the selected prefecture, 86.1% (539/626) of them were infected through commercial plasma donation or clinically operated blood transfusion. With respect to disease surveillance and identification in the area, 52.2% (327/626) of all the cases were reported by hospitals or clinics. The number and proportion of HIV/AIDS cases identified or reported at county, prefecture, and provincial levels were 207 (33.1%), 303 (48.4%) and 116 (18.5%), respectively. The number and proportion of HIV/AIDS identified through specific epidemiological investigations and/or voluntary testing had been increased in recent years. In addition, among HIV/AIDS cases that were clinically identified, the proportion of those who were identified as outpatients had steadily increased,with the highest proportion (59.3%) observed in 2004., Conclusions: In rural areas of central China where the major mode of HIV transmission was through commercial plasma donation or clinically operated blood transfusion, hospitals and clinics seemed to have played and would continue to play important roles regarding HIV identification and surveillance. The role of institutions or settings at the prefecture level regarding HIV identification and surveillance should not be ignored.

Objective: To investigate the feasibility and risk of preimplantation genetic diagnosis (PGD) for screening normal offspring of Robertsonian translocation carriers., Methods: This case was clinically diagnosed as primary infertility for 6 years; the husband was found to have chromosome der (13;14) (q10;q10) and oligozoospermia. For the solution of the couple's problem, controlled ovarian hyperstimulation (COH) and intracytoplasmic sperm injection (ICSI) were performed to obtain embryos. The embryos were drilled in zona by acidified Tyrode's solution at 6-8 cell stage (day 3 post-fertilization) and a single blastomere was removed from each embryo. All blastomeres were analyzed by fluorescence in situ hybridization (FISH) using the double color probes LSI 13q labeled by SpectrumOrange and Tel 14q labeled by SpectrumGreen. The embryos biopsied were cultured at once and the normal ones selected were transferred the next day. Prenatal diagnostic techniques were used to detect the karyotype of fetus at 18 weeks of gestation., Results: Unbalanced, normal or balanced, and unclear embryos were separated. The couple obtained 50a (4/8)normal or balanced,and 37.5a (3/8)unbalanced, and 12.5a (1/8) unclear embryos. A singleton pregnancy followed, and the karyotype of the fetus (46,XY) was detected by prenatal diagnostic techniques., Conclusion: PGD is useful for screening out unbalanced embryos and is very valuable for solving the reproductive problem of Robertsonian translocation carriers and for avoiding fetal beings with severe disorders.

Analysis of two exopolysaccharide-deficient mutants of Rhizobium leguminosarum, RBL5808 and RBL5812, revealed independent Tn5 transposon integrations in a single gene, designated exo5. As judged from structural and functional homology, this gene encodes a UDP-glucose dehydrogenase responsible for the oxidation of UDP-glucose to UDP-glucuronic acid. A mutation in exo5 affects all glucuronic acid-containing polysaccharides and, consequently, all galacturonic acid-containing polysaccharides. Exo5-deficient rhizobia do not produce extracellular polysaccharide (EPS) or capsular polysaccharide (CPS), both of which contain glucuronic acid. Carbohydrate composition analysis and nuclear magnetic resonance studies demonstrated that EPS and CPS from the parent strain have very similar structures. Lipopolysaccharide (LPS) molecules produced by the mutant strains are deficient in galacturonic acid, which is normally present in the core and lipid A portions of the LPS. The sensitivity of exo5 mutant rhizobia to hydrophobic compounds shows the involvement of the galacturonic acid residues in the outer membrane structure. Nodulation studies with Vicia sativa subsp. nigra showed that exo5 mutant rhizobia are impaired in successful infection thread colonization. This is caused by strong agglutination of EPS-deficient bacteria in the root hair curl. Root infection could be restored by simultaneous inoculation with a Nod factor-defective strain which retained the ability to produce EPS and CPS. However, in this case colonization of the nodule tissue was impaired.

Objective: To explore the possibility of phenotypic conversion of clinical chloromycetin (Cm)-resistant isolates of E.coli to drug-sensitive ones with external guide sequences (EGS) in vitro., Methods: Recombinant EGS plasmids directed against Cm acetyl transferase (cat) and containing kanamycin (Km) drug-resistance gene and control plasmids only containing kanamycin-resistance gene without EGS were constructed. By using CaCl(2) method, the recombinant plasmids were introduced into the clinically isolated Cm-resistant E.coli strains. Extraction of plasmids and PCR were applied to identify the EGS positive clones; The growth rate in liquid broth culture of Cm-resistant bacteria after EGS containing plasmid transformation was determined by spectrophotometer A(600). Drug sensitivity was tested in solid culture by using KB method., Results: Transformation studies were carried out on 16 clinically isolated Cm-resistant E.coli strains with pEGFP-C1-EGS + cat1 + cat2 recombinant plasmid. Transformants were screened on LB-agar plates containing Km after transformation using EGS. In 4 tested strains of them, transformants with specific EGS plasmid showed growth inhibition when grown in liquid broth culture containing 100 approximately 200 micro g/ml of Cm. They were sensitive to Cm on LB-agar plates containing 100 approximately 200 micro g/ml of Cm in drug-sensitivity test. Extraction of plasmids showed the existence of EGS bands. PCR amplified products of EGS. The above facts indicated that the 4 strains out of the 16 clinical isolates had been converted to drug-sensitive phenotype, and Cm-resistant clinically isolated E. coli resumed sensitivity to Cm., Conclusion: EGS has the capability of converting the phenotype of clinical drug-resistant isolates to drug sensitivity.

Objective: To evaluate the reliability of different hepatitis E diagnosis reagent tests on the acute hepatitis E., Methods: Three acute hepatitis E diagnosis tests, E2-IgM (Wantai, China), GL-IgM and GL-IgG (Genelabs, Singapore) were compared for their reliability in a sera panel composed by 273 healthy individuals and 525 hepatitis., Results: The specificity of E2-IgM on the diagnosis of acute hepatitis E was 100.0%, it was significantly higher than GL-IgM (96.7%) and GL-IgG (85.4%). The sensitivity of E2-IgM and GL-IgG were 97.9% and 93.8% respectively, both significantly higher than GL-IgM (72.9%). Among 65 acute hepatitis cases being positive on GL-IgM test but negative on E2-IgM, 58 (89.2%) cases were found to be positive with anti-hepatitis A virus IgM, it indicated that the GL-IgM test might be interfered by other IgM antibodies on serum., Conclusion: E2-IgM is a good test for the diagnosis of acute hepatitis E.

The elucidation of the structure of the O-antigen of Rhizobium etli CE3 predicts that the R. etli CE3 genome must contain genes encoding acetyl- and methyltransferases to confer the corresponding modifications to the O-antigen. We identified three open reading frames (ORFs) upstream of wzm, encoding the membrane component of the O-antigen transporter and located in the lps alpha-region of R. etli CE3. The ORFs encode two putative acetyltransferases with similarity to the CysE-LacA-LpxA-NodL family of acetyltransferases and one putative methyltransferase with sequence motifs common to a wide range of S-adenosyl-L-methionine-dependent methyltransferases. Mutational analysis of the ORFs encoding the putative acetyltransferases and methyltransferase revealed that the acetyl and methyl decorations mediated by these specific enzymes are essential for O-antigen synthesis. Composition analysis and high performance anion exchange chromatography analysis of the lipopolysaccharides (LPSs) of the mutants show that all of these LPSs contain an intact core region and lack the O-antigen polysaccharide. The possible role of these transferases in the decoration of the O-antigen of R. etli is discussed.

The synthesis of 2,3-dideoxy-3-C-(hydroxymethyl)-3-C-methyl-D-glycero-tetrofuranosyl++ + nucleosides was accomplished in high enatiomeric purity (98.5% ee) via [3,3]-sigmatropic Claisen rearrangement of (E)(S)-5-benzyloxy-1-tert-butyldimethylsilanyloxy-4-methyl-pent-3- en-2-ol prepared from 2,3-O-isopropylidene-D-glyceraldehyde. The synthesized nucleosides were assayed against human immunodeficiency virus (HIV) and hepatitis B virus in human peripheral blood mononuclear (PBM) and 2.2.15 cells, respectively. 6-Amino-9-[2,3-dideoxy-3-C-(hydroxymethyl)-3-C-methyl-beta-D-glycero- tetrofuranosyl]-2-fluoropurine shows moderate antiviral activity (EC50 = 2.55 microM) against HIV-1 strains and 6-amino-9-[3-deoxy-3-C-(hydroxymethyl)-3-methyl-alpha-D-glycero-tetro furanosyl]-2-fluoropurine exhibits potent anti-HIV activity (EC50 = 0.073 microM) with significant cytotoxicity (IC50 = 1.0 microM).

Objectives: to test whether the occurrence of microembolism differed between eversion and standard carotid endarterectomy (CEA)., Design: prospective, non-randomised transcranial Doppler (TCD) monitoring study of 61 patients., Materials and Methods: eversion CEA was performed in 27 and standard CEA in 34 patients. Surgery was performed under general anaesthesia. Three (5%) patients had a shunt inserted based on continuous EEG monitoring. Continuous middle cerebral artery TCD monitoring (EME, TC-4040) was performed intraoperatively and for 45 min postoperatively on day 1, day 2-3, day 4-5 and after 3 months. Unidirectional signals lasting >25 ms, having intensities of >9 dB were considered to represent embolic events., Results: intraoperative embolic events were detected in 50 (93%) of 54 patients in whom successful intraoperative TCD monitoring was achieved. Events occurred most frequently immediately following clamp release (85%), without difference between the two techniques. Embolic events were encountered postoperatively in four (15%) and 16 (48%) patients having eversion and standard CEA, respectively (p< 0.02). Four patients developed new neurological symptoms equally distributed between eversion and standard CEA. Two (3%) deficits were permanent and two transient. The patency of the carotid bifurcation was confirmed in all instances with duplex scanning., Conclusion: we identified a surprisingly high number of postoperative embolic events as detected with transcranial Doppler in patients who had undergone carotid surgery using standard endarterectomy, as compared to patients who underwent eversion endarterectomy. Whether this difference has any clinical implication has yet to be proved., (Copyright 2000 Harcourt Publishers Ltd.)

China's dual employment system plays a crucial role in sexually transmitted infections (STIs) and HIV-related safe and unsafe sexual practices among young Chinese people. Social and psychological determinants of safe and unsafe sexual practices for HIV infection among young people in Sichuan, China were examined. Our findings indicate that changes in China's social structure and employment system impact upon the social contextual involvement and socio-sexual practice of young Chinese people. The findings in the study suggest that the employment-related contextual involvement was a major predictor in the relationships between demography, information, and psychological risk-taking factors on one hand and the people's safe and unsafe sexual practices on the other. Self-employed people (officially called 'getihu') were more likely than the state-employed people to engage in unprotected sex with casual sexual partners. As China undergoes social restructuring and many state-employed people are laid off, the risk may also extend into the broader non-self-employed population as more state-employed people become involved not only in the self-employed getihu's socioeconomic activities but also in their unconventional socio-sexual practices. Collective vulnerability to STI and HIV, due to the current socio-sexual practices of the getihu young people, has created a new frontier for STI and HIV prevention in today's China, as well as demonstrating the importance of collective action with STI and AIDS prevention strategies within relevant social and sub-cultural contexts.