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Michael Guccione,Sharon Silbiger,Jun Lei,Joel Neugarten The accumulation of extracellular matrix in the glomerular mesangium reflects the net balance between the synthesis and degradation of matrix components. We have shown that estradiol suppresses the synthesis of types I and IV collagen by cultured mesangial cells (Kwan G, Neugarten J, Sherman M, Din... ( view more )g Q, Fotadar U, Lei J, and Silbiger S. Kidney Int 50: 1173-1179, 1996; Neugarten J, Acharya A, Lei J, and Silbiger S. Am J Physiol Renal Physiol 279: F309-F318, 2000; Neugarten J, Medve I, Lei J, and Silbiger SR. Am J Physiol Renal Physiol 277: F1-F8, 1999; Neugarten J and Silbiger S. Am J Kidney Dis 26: 147-151, 1995; Silbiger S, Lei J, and Neugarten J. Kidney Int 55: 1268-1276, 1998; Silbiger S, Lei J, Ziyadeh FN, and Neugarten J. Am J Physiol Renal Physiol 274: F1113-F1118, 1998). In the present study, we evaluated the effects of sex hormones on the activity of matrix metalloproteinase-2 (MMP-2) in murine mesangial cells, the synthesis of which is regulated by the transcription factor activator protein-2 (AP-2). Estradiol stimulated MMP-2 activity by increasing MMP-2 protein levels in a dose-dependent manner. These effects occurred at physiological concentrations of estradiol and were receptor mediated. Estradiol also increased AP-2 protein levels and increased binding of mesangial cell nuclear extracts to an AP-2 consensus binding sequence oligonucleotide. The ability of estradiol to increase AP-2 protein expression, AP-2/DNA binding activity, MMP-2 protein expression, and metalloproteinase activity was reversed by PD-98059, a selective inhibitor of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling cascade. We conclude that estradiol upregulates the MAPK cascade, which in turn stimulates the synthesis of AP-2 protein. The resultant increased AP-2/DNA binding activity leads to increased synthesis of MMP-2 and increased metalloproteinase activity. Stimulation of metalloproteinase activity by estradiol may contribute to the protective effect of female gender on renal disease progression. ( view less ) S Morris,J Leis Predicted secondary-structure elements encompassing the primer binding site in the 5' untranslated region of Rous sarcoma virus (RSV) RNA play an integral role in multiple viral replications steps including reverse transcription, DNA integration, and RNA packaging (A. Aiyar, D. Cobrinik, Z. Ge, H. ... ( view more )J. Kung, and J. Leis, J. Virol. 66:2464-2472, 1992; D. Cobrinik, A. Aiyar, Z. Ge, M. Katzman, H. Huang, and J. Leis, J. Virol. 65:3864-3872, 1991; J. T. Miller, Z. Ge, S. Morris, K. Das, and J. Leis, J. Virol. 71:7648-7656, 1997). These elements include the U5-Leader stem, U5-IR stem-loop, and U5-TPsiC interaction region. Limited digestion of the 5' untranslated region of wild-type and mutant RSV RNAs with structure- and/or sequence-specific RNases detects the presence of the U5-Leader stem and the U5-IR stem-loop. When a tRNATrp primer is annealed to wild-type RNAs in vitro, limited nuclease mapping indicates that the U5-IR stem becomes partially unwound. This is not observed when mutant RNAs with altered U5-IR stem-loop structures are substituted for wild-type RNAs. The U5-Leader stem also becomes destabilized when the tRNA primer is annealed to either wild-type or mutant RNA fragments. Nuclease mapping studies of tRNATrp, as well as the viral RNA, indicate that the U5-TPsiC helix does form in vitro upon primer annealing. Collectively, these data suggest that the various structural elements near the RSV primer binding site undergo significant changes during the process of primer annealing. ( view less ) Lei Zhang,Yong Zhao,Lei Zhang,Ting Liu,Chun-Ying Li,Ran Jin,Lei Zhou,Jian-Rong Li OBJECTIVE: To discuss the protective effect of Niuhuang Shangqing pills (NSP) on the experimental cerebral ischemia and the influence of blood rheology in animals. METHOD: Using the middle cerebral artery occlusion (MCAO), bilateral common carotid artery ligation and lipopolysaccharide (LPS) in com... ( view more )bination with carrogeenin (Ca)-induced stagnation of blood model rats, we investigated the influence of NSP on the physical sign indexes, brain infarct size and the water content, the content of lactate(LD), glutathione peroxidase (GPx), catalase (CAT), malondialdehyde (MDA) and the blood rheology. RESULT: The 3.00, 1.50 g x kg(-1) dosage groups of NSP decreased the neurosigns indexes, significantly reduced the brain infarct areas in MCAO rats, decreased the brain water content, increased the activities of GPx and CAT, decreased the content of LD and MDA in bilateral common carotid artery ligation rats. They improved the blood rheology in lipopolysaccharide (LPS) in combination with carrogeenin (Ca)-induced stagnation of blood model rats. CONCLUSION: NSP has the function of protective effect on experimental experimental cerebral ischemia. ( view less ) Jun Yu,Jun Wang,Wei Lin,Songgang Li,Heng Li,Jun Zhou,Peixiang Ni,Wei Dong,Songnian Hu,Changqing Zeng,Jianguo Zhang,Yong Zhang,Ruiqiang Li,Zuyuan Xu,Shengting Li,Xianran Li,Hongkun Zheng,Lijuan Cong,Liang Lin,Jianning Yin,Jianing Geng,Guangyuan Li,Jianping Shi,Juan Liu,Hong Lv,Jun Li,Jing Wang,Yajun Deng,Longhua Ran,Xiaoli Shi,Xiyin Wang,Qingfa Wu,Changfeng Li,Xiaoyu Ren,Jingqiang Wang,Xiaoling Wang,Dawei Li,Dongyuan Liu,Xiaowei Zhang,Zhendong Ji,Wenming Zhao,Yongqiao Sun,Zhenpeng Zhang,Jingyue Bao,Yujun Han,Lingli Dong,Jia Ji,Peng Chen,Shuming Wu,Jinsong Liu,Ying Xiao,Dongbo Bu,Jianlong Tan,Li Yang,Chen Ye,Jingfen Zhang,Jingyi Xu,Yan Zhou,Yingpu Yu,Bing Zhang,Shulin Zhuang,Haibin Wei,Bin Liu,Meng Lei,Hong Yu,Yuanzhe Li,Hao Xu,Shulin Wei,Ximiao He,Lijun Fang,Zengjin Zhang,Yunze Zhang,Xiangang Huang,Zhixi Su,Wei Tong,Jinhong Li,Zongzhong Tong,Shuangli Li,Jia Ye,Lishun Wang,Lin Fang,Tingting Lei,Chen Chen,Huan Chen,Zhao Xu,Haihong Li,Haiyan Huang,Feng Zhang,Huayong Xu,Na Li,Caifeng Zhao,Shuting Li,Lijun Dong,Yanqing Huang,Long Li,Yan Xi,Qiuhui Qi,Wenjie Li,Bo Zhang,Wei Hu,Yanling Zhang,Xiangjun Tian,Yongzhi Jiao,Xiaohu Liang,Jiao Jin,Lei Gao,Weimou Zheng,Bailin Hao,Siqi Liu,Wen Wang,Longping Yuan,Mengliang Cao,Jason McDermott,Ram Samudrala,Jian Wang,Gane Ka-Shu Wong,Huanming Yang We report improved whole-genome shotgun sequences for the genomes of indica and japonica rice, both with multimegabase contiguity, or almost 1,000-fold improvement over the drafts of 2002. Tested against a nonredundant collection of 19,079 full-length cDNAs, 97.7% of the genes are aligned, without ... ( view more )fragmentation, to the mapped super-scaffolds of one or the other genome. We introduce a gene identification procedure for plants that does not rely on similarity to known genes to remove erroneous predictions resulting from transposable elements. Using the available EST data to adjust for residual errors in the predictions, the estimated gene count is at least 38,000-40,000. Only 2%-3% of the genes are unique to any one subspecies, comparable to the amount of sequence that might still be missing. Despite this lack of variation in gene content, there is enormous variation in the intergenic regions. At least a quarter of the two sequences could not be aligned, and where they could be aligned, single nucleotide polymorphism (SNP) rates varied from as little as 3.0 SNP/kb in the coding regions to 27.6 SNP/kb in the transposable elements. A more inclusive new approach for analyzing duplication history is introduced here. It reveals an ancient whole-genome duplication, a recent segmental duplication on Chromosomes 11 and 12, and massive ongoing individual gene duplications. We find 18 distinct pairs of duplicated segments that cover 65.7% of the genome; 17 of these pairs date back to a common time before the divergence of the grasses. More important, ongoing individual gene duplications provide a never-ending source of raw material for gene genesis and are major contributors to the differences between members of the grass family. ( view less ) Lin Yuan,Lei Tang,Wen-hua Huang,Jian-yi Li,Jin-xing Dai,Chang Liu,Tao Wu,Xing-hai Wan,Hui-wen Hong,Mei-chao Zhang,Pei-feng Jiao,Yun-tao Lu,Kun-cheng Wu,Pei-liang Li,Ji-hong Fan,Yuan Gao,Qing-zhi Wang,Long-jiang Wang,Lei Wu,Lei Zhang,Xin-an Li,Ying-hua Chen,Jun Ouyang,Shi-zhen Zhong OBJECTIVE: To establish digitized Virtual Chinese Human Male No.1 (VCH-M1) image dataset with a 0.2-mm equal interval. METHODS: The body of a 24-year-old male was used for this study. Perfusion with phenol and vermilion of the arteries was performed, followed by body shape adjustment by cold saline... ( view more ) and pre-embedding with broken ices in an upside-down position, which was completed in a stepwise procedure to minimize body shape deformation. Section milling was conducted subsequently with the section thickness of 2 mm and the section images were captured by digital camera, which were immediately transferred to a computer for storage and processing. RESULTS: A total of 9 232 sections were obtained for the whole body, and the resolution of each of the image in TIF format was 3 024x2 016 pixels, resulting in the size of approximately 18 M for each image and about 161 G for the whole dataset. CONCLUSIONS: Compared with VCH-F1, the image quality in VCH-M1 dataset is significantly improved, demonstrated by much clearer tissue boundary in the images and minimized body shape deformation during the embedding process. ( view less ) Qi Feng,Yujun Zhang,Pei Hao,Shengyue Wang,Gang Fu,Yucheng Huang,Ying Li,Jingjie Zhu,Yilei Liu,Xin Hu,Peixin Jia,Yu Zhang,Qiang Zhao,Kai Ying,Shuliang Yu,Yesheng Tang,Qijun Weng,Lei Zhang,Ying Lu,Jie Mu,Yiqi Lu,Lei S Zhang,Zhen Yu,Danlin Fan,Xiaohui Liu,Tingting Lu,Can Li,Yongrui Wu,Tongguo Sun,Haiyan Lei,Tao Li,Hao Hu,Jianping Guan,Mei Wu,Runquan Zhang,Bo Zhou,Zehua Chen,Ling Chen,Zhaoqing Jin,Rong Wang,Haifeng Yin,Zhen Cai,Shuangxi Ren,Gang Lv,Wenyi Gu,Genfeng Zhu,Yuefeng Tu,Jia Jia,Yi Zhang,Jie Chen,Hui Kang,Xiaoyun Chen,Chunyan Shao,Yun Sun,Qiuping Hu,Xianglin Zhang,Wei Zhang,Lijun Wang,Chunwei Ding,Haihui Sheng,Jingli Gu,Shuting Chen,Lin Ni,Fenghua Zhu,Wei Chen,Lefu Lan,Ying Lai,Zhukuan Cheng,Minghong Gu,Jiming Jiang,Jiayang Li,Guofan Hong,Yongbiao Xue,Bin Han Rice is the principal food for over half of the population of the world. With its genome size of 430 megabase pairs (Mb), the cultivated rice species Oryza sativa is a model plant for genome research. Here we report the sequence analysis of chromosome 4 of O. sativa, one of the first two rice chrom... ( view more )osomes to be sequenced completely. The finished sequence spans 34.6 Mb and represents 97.3% of the chromosome. In addition, we report the longest known sequence for a plant centromere, a completely sequenced contig of 1.16 Mb corresponding to the centromeric region of chromosome 4. We predict 4,658 protein coding genes and 70 transfer RNA genes. A total of 1,681 predicted genes match available unique rice expressed sequence tags. Transposable elements have a pronounced bias towards the euchromatic regions, indicating a close correlation of their distributions to genes along the chromosome. Comparative genome analysis between cultivated rice subspecies shows that there is an overall syntenic relationship between the chromosomes and divergence at the level of single-nucleotide polymorphisms and insertions and deletions. By contrast, there is little conservation in gene order between rice and Arabidopsis. ( view less ) Qiang Zhao,Yu Zhang,Zhukuan Cheng,Mingsheng Chen,Shengyue Wang,Qi Feng,Yucheng Huang,Ying Li,Yesheng Tang,Bo Zhou,Zhehua Chen,Shuliang Yu,Jingjie Zhu,Xin Hu,Jie Mu,Kai Ying,Pei Hao,Lei Zhang,Yiqi Lu,Lei S Zhang,Yilei Liu,Zhen Yu,Danlin Fan,Qijun Weng,Ling Chen,Tingting Lu,Xiaohui Liu,Peixin Jia,Tongguo Sun,Yongrui Wu,Yujun Zhang,Ying Lu,Can Li,Rong Wang,Haiyan Lei,Tao Li,Hao Hu,Mei Wu,Runquan Zhang,Jianping Guan,Jia Zhu,Gang Fu,Minghong Gu,Guofan Hong,Yongbiao Xue,Rod Wing,Jiming Jiang,Bin Han As part of an international effort to completely sequence the rice genome, we have produced a fine bacterial artificial chromosome (BAC)-based physical map of the Oryza sativa japonica Nipponbare chromosome 4 through an integration of 114 sequenced BAC clones from a taxonomically related subspecies... ( view more ) O. sativa indica Guangluai 4 and 182 RFLP and 407 expressed sequence tag (EST) markers with the fingerprinted data of the Nipponbare genome. The map consists of 11 contigs with a total length of 34.5 Mb covering 94% of the estimated chromosome size (36.8 Mb). BAC clones corresponding to telomeres, as well as to the centromere position, were determined by BAC-pachytene chromosome fluorescence in situ hybridization (FISH). This gave rise to an estimated length ratio of 5.13 for the long arm and 2.9 for the short arm (on the basis of the physical map), which indicates that the short arm is a highly condensed one. The FISH analysis and physical mapping also showed that the short arm and the pericentromeric region of the long arm are rich in heterochromatin, which occupied 45% of the chromosome, indicating that this chromosome is likely very difficult to sequence. To our knowledge, this map provides the first example of a rapid and reliable physical mapping on the basis of the integration of the data from two taxonomically related subspecies. ( view less ) A Aiyar,Z Ge,J Leis The 5' end of avian retrovirus RNA near the primer-binding site (PBS) forms two secondary structures, the U5-inverted repeat (U5-IR) and the U5-leader stems, and contains a 7-nucleotide sequence that anneals to the T psi C loop of the tRNA(Trp) primer. Mutations that disrupt any of these base pair ... ( view more )interactions cause defects in initiation of reverse transcription both in vivo and in vitro (D. Cobrinik, A. Aiyar, Z. Ge, M. Katzman, H. Huang, and J. Leis, J. Virol. 65:3864-3872, 1991; A. Aiyar, D. Cobrinik, Z. Ge, H.-J. Kung, and J. Leis, J. Virol. 66:2464-2472, 1992). We have now examined the effect of perturbing the non-base-paired intervening "spacer" sequences between these secondary-structure elements. Small deletions or insertions in these intervening sequences decreased initiation of reverse transcription in vitro. In contrast, base substitutions, which maintain the spacing distances between the structures, had no detectable effect. Additionally, a small deletion at the 3' end of the PBS caused a significant decrease in initiation of reverse transcription whereas substitution mutations again had no effect. Together, these results indicate that reverse transcriptase forms a complex in which the different structural elements are maintained in a specific orientation that is required for efficient initiation of reverse transcription. Specific sequence recognition of the duplex structures by reverse transcriptase is also required since mosaic RNAs that combine the human immunodeficiency virus type 1 PBS with avian sequences is not efficiently utilized for reverse transcription even though the primer used can anneal to the substituted PBS. ( view less ) Lei Cheng,Liang Zhou,Lei Tao,Ming Zhang,Jiefeng Cui,Yan Li OBJECTIVES: Proteomic profiling of serum is an emerging technique to identify new biomarkers indicative of disease severity and progression. Our study was to assess the use of surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) to identify multiple serum pro... ( view more )tein biomarkers for early detection of laryngeal squamous cell carcinoma (LSCC), establish predictive model, and accurately distinguish LSCC patients with or without lymph node metastasis. METHODS: A cohort of 252 serum samples with LSCC (n = 142) and normal control (n = 110) were consented into this study. These serum samples were randomly divided into training set (including 89 LSCC patients at stages I-II and 65 normal controls, 30 LSCC patients with lymph node metastasis) and blind testing set (including 53 LSCC patients at stages III-IV and 45 normal controls). Serum protein profiles on weak cationic exchange (WCX2) were performed by SELDI-TOF MS and then analyzed by Biomarker Wizard software. The Decision Tree classification algorithm and blind validation were determined by Biomarker Pattern Software (BPS). RESULTS: A panel of 18 biomarkers ranging 2-30 kDa was selected based on their collective contribution to the optimal separation between stages I-II LSCC patients and healthy controls. Among them, one candidate protein peak with an m/z value of 4,176 Da was selected to establish predictive model by BPS with sensitivity of 86.52% and specificity of 84.62%. The ability to detect LSCC patients was evaluated using blinding test data in stages III and IV cancer patients. A sensitivity of 84.91% and specificity of 82.22% were validated in blind testing set. Meanwhile 14 potential biomarkers could differentiate LSCC patients with or without lymph node metastasis (P < 0.05). CONCLUSIONS: The high sensitivity and specificity achieved by the serum protein biomarkers show great potential for the early detection of LSCC. SELDI-TOF MS serum profiling also is able to distinguish LSCC patients with or without lymph node metastasis. ( view less ) Bei-Wei Zhu,Lu-Lu Zhao,Li-Ming Sun,Dong-Mei Li,Yoshiyuki Murata,Lei Yu,Lei Zhang Cathepsin L-like enzyme was purified from the body wall of the sea cucumber Stichopus japonicus by an integral method involving ammonium sulfate precipitation and a series of column chromatographies on DEAE Sepharose CL-6B, Sephadex G-75, and TSK-GEL. The molecular mass of the purified enzyme was e... ( view more )stimated to be 63 kDa by SDS-PAGE. The enzyme cleaved N-carbobenzoxy-phenylalanine-arginine7-amido-4-methylcoumarin with K(m) (69.92 microM) and k(cat) (12.80/S) hardly hydrolyzed N-carbobenzoxy-arginine-arginine 7-amido-4-methylcoumarin and L-arginine 7-amido-4-methylcoumarin. The optimum pH and temperature for the purified enzyme were found to be 5.0 and 50 degrees C. It showed thermal stability below 40 degrees C. The activity was inhibited by sulfhydryl reagents and activated by reducing agents. These results suggest that the purified enzyme was a cathepsin L-like enzyme and that it existed in the form of its enzyme-inhibitor complex or precursor. ( view less ) Ping Liu,Ji-zhou Xiang,Lei Zhao,Lei Yang,Ben-rong Hu,Qin Fu AIM: To observe the effect of beta2-adrenergic agonist clenbuterol on ischemia/reperfusion (I/R) injury in isolated rat hearts and hydrogen peroxide (H2O2)-induced cardiomyocyte apoptosis. METHODS: Isolated rat hearts were subjected to 30 min global ischemia and 60 min reperfusion on a Langendorff ... ( view more )apparatus. Cardiac function was evaluated by heart rate, left ventricular end-diastolic pressure (LVEDP), left ventricular systolic pressure, maximal rise rate of left ventricular pressure [+dp/dt(max)], and the coronary effluent (CF). Lactate dehydrogenase (LDH) in the coronary effluent, malondialdehyde (MDA), superoxide dismutase (SOD), and Ca2+-ATPase activity in the cardiac tissue were measured using commercial kits. The apoptotic cardiomyocyte was detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay. Bax/Bcl-2 mRNA levels and the expression of caspase-3 were detected by RT-PCR and immunoblotting, respectively. Cultured newborn rat cardiomyocytes were preincubated with clenbuterol, and oxidative stress injury was induced by H2O2. Cell viability and cardiomyocyte apoptosis were evaluated by flow cytometry (FCM). RESULTS: In the isolated rat hearts after I/R injury, clenbuterol significantly improved diastolic function (LVEDP and CF) and Ca2+-ATPase activity. Treatment with clenbuterol increased SOD activity and decreased the MDA level and LDH release compared with the I/R group (P<0.05). Moreover, clenbuterol decreased apoptosis, which was associated with a reduction in TUNEL-positive cells, Bax/Bcl-2 mRNA, and caspase-3 expression. In H2O2-induced cardiomyocyte injury, clenbuterol increased cell viability and attenuated cardiomyocyte apoptosis. Pretreatment with ICI118551 (selective beta2-adrenergic antagonist) decreased these effects compared with the clenbuterol-treated group (P<0.05). CONCLUSION: Clenbuterol ameliorated ventricular diastolic function by enhancing Ca2+-ATPase activity and reduced oxidative stress and cardiac myocyte apoptosis in an experimental rat model of myocardium I/R. It decreased cardiomyocyte apoptosis induced by H2O2 in vitro. It plays a key role in the cardiac protection against myocardium I/R injury. ( view less ) Lei Shang,Liying Liu,Lei XuSingle-mode lasing from a coupled asymmetric microcavity is achieved. By coupling two size mismatched circular microrings to form a coupled asymmetric microcavity, multi-whispering-gallery modes are successfully suppressed and single-frequency laser emission is robustly obtained. Moreover, the lase... ( view more )r emits in four directions, and each beam has a divergence of only 6.6 degrees . It is demonstrated further that this single-frequency coupled microcavity laser can be easily integrated with planar lightwave circuits. We provide an easily accessible approach to achieve a single-frequency laser from microcavity lasers operating on whispering-gallery modes. ( view less ) Jing Zhou,Wei Lei,Lei Shen,He-Sheng Luo,Zhi-Xiang Shen AIM: To investigate the expression level and effects of leptin in human hepatocellular carcinoma cells in vitro and to explore the correlation between them. METHODS: Human hepatocellular carcinoma cell line HepG2 was cultured in vitro, and (the expression level) mRNA of leptin and leptin receptors ... ( view more )in HepG2 were assessed using reverse transcription polymerase chain reaction (RT-PCR). Effects of different concentrations of leptin (50 ng/mL, 100 ng/mL, 200 ng/mL) on HepG2 were detected with colorimetric assay by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) after incubation periods of 24 h, 48 h, and 72 h. Flow cytometry was performed to assess cell cycle progression of different concentrations of leptin as stated above after each 24 h incubation period. RESULTS: mRNA of leptin and leptin receptors (including short and long isoforms) were expressed in HepG2. The 72 h incubation of leptin at different concentrations (50 ng/mL, 100 ng/mL, 200 ng/mL) promoted proliferation of HepG2 in a concentration- and time-dependent manner. The experimental group shows significant statistical differences when compared to the controlled group which contained 0 ng/mL of leptin. As the concentration of leptin increases, significant fewer cells were detected in G0-G1 phase and more cells in S and G2-M phases. CONCLUSION: Leptin and leptin receptor are simultaneously expressed in human hepatocellular carcinoma cell line HepG2. Addition of leptin (0 ng/mL-200 ng/mL) in 72 h periods indicated there is a concentration- and time-dependent correlation in the stimulation of HepG2 cell proliferation. The effect of proliferation by leptin is due to promotion of DNA synthesis and enhancement of mitotic activity. The relationship between leptin and human hepatocellular carcinoma cells might indicate that adipokine could be associated with the progression of human hepatocellular carcinoma. ( view less ) Lei Shi,Wenbin Yue,Youshe Ren,Fulin Lei,Junxing Zhao The objective of this study was to obtain a fast, accurate and reliable method of determining the sex of goat embryos prior to implantation through amplification of the high-motility-group (HMG) box of the sex-determining region of the Y chromosome (SRY) gene of the goats. Goat specific primers wer... ( view more )e designed for duplex polymerase chain reaction (PCR). As an internal control gene, the goat beta-action gene sequence was simultaneously amplified together with the HMG box of goat SRY gene. Males showed both 1 SRY band and 1 beta-action band, but only 1 beta-action band was present in the agarose gel electrophoresis of females. The result indicated that the goat HMG-box sequence motif of SRY was male specific. Afterward, the optimized PCR procedure was applied in 30 embryo biopsies and the biopsied embryos were transferred into 30 recipient female goats. The sex of the 13 kids proved anatomically corresponded to the sex determined by PCR (100% accuracy). Thus, this study showed that this duplex PCR method can be applied to sex the goat pre-implantation embryos and to manipulate the sex ratio of offspring in goat breeding programs. ( view less ) Gang Guo,Lei Zhang,Zhou Zhou,Qiqi Ma,Jianping Liu,Chenguang Zhu,Lei Zhu,Ziniu Yu,Ming Sun Bacillus thuringiensis produces various groups of active proteins, such as Cyt, Vip and Parasporin, in addition to the Cry protein. In this study we show S-layer proteins to be a new group of parasporal inclusions of B. thuringiensis. The S-layer consists of a two-dimensional lattice structure and ... ( view more )is the outermost component of many archaeobacteria and eubacteria. The parasporal inclusion of B. thuringiensis strain CTC was found to be not a typical crystal protein encoded by the cry gene, but a proteinaceous inclusion encoded by the S-layer gene. Furthermore, the CTC-like strains (with their parasporal inclusions coded by the S-layer gene) are widely distributed and accounted for 25.4% of the B. thuringiensis strains tested. These strains constitute a new group of parasporal inclusions encoded by the S-layer gene of B. thuringiensis and shed new light on B. thuringiensis nontoxic strains. ( view less ) Lei Huo,Lei Wu,Ming-Yu Zhang,Yong-Hong Hou,Xi-Ping Ding,Jia-Sheng FangObjective To investigate the curative effect of electrocorticography (ECoG) monitoring in the microsurgical treatment of cavernous angiomas.Methods Clinical data of 71 patients with epileptogenic cavernous angiomas,who had been performed ECoG monitoring during the operation,were analyzed retrospect... ( view more )ively.Results The foci of cavernous angiomas and epilepsy of the 71 patients were resected during the operation. In the 58 patients who were followed up,42 had not epileptic seizure,and 16 still had epileptic seizure,while the frequencies of 13 patients reduced to below 10%,and 3 patients over 10%. Conclusion The drug treatment of epileptogenic cavernous angiomas can not control epileptic seizure,and the patients should receive the microsurgical treatment early. Electrocorticography monitoring can direct the surgical procedure,and control the postoperative epileptic seizure. ( view less ) Zhi-Yan Shan,Jing-Ling Shen,Qiu-Ming Li,Yan Wang,Xiao-Yi Huang,Tie-Yun Guo,Hui-Wen Liu,Lei Lei,Lian-Hong JinMouse embryonic stem (ES) cells can be induced by various chemicals to differentiate into a variety of cell types in vitro. In our study, retinoic acid (RA), one of the most important inducers, used at a concentration of 5 microM, was found to induce the differentiation of ES cells into neural prog... ( view more )enitor cells (NPCs). During embryoid body (EB) differentiation, the level of active cyclic AMP response element-binding protein (CREB) was relatively high when 5 microM RA treatment was performed. Inhibition of CREB activity committed EBs to becoming other germ layers, whereas increased expression of CREB enhanced NPC differentiation. Moreover, RA increased the expression of active CREB by enhancing the activity of JNK. Our research suggests that CREB plays a role in RA-induced NPC differentiation by increasing the expression of active JNK. ( view less ) Ray Ming,Shaobin Hou,Yun Feng,Qingyi Yu,Alexandre Dionne-Laporte,Jimmy H Saw,Pavel Senin,Wei Wang,Benjamin V Ly,Kanako L T Lewis,Steven L Salzberg,Lu Feng,Meghan R Jones,Rachel L Skelton,Jan E Murray,Cuixia Chen,Wubin Qian,Junguo Shen,Peng Du,Moriah Eustice,Eric Tong,Haibao Tang,Eric Lyons,Robert E Paull,Todd P Michael,Kerr Wall,Danny W Rice,Henrik Albert,Ming-Li Wang,Yun J Zhu,Michael Schatz,Niranjan Nagarajan,Ricelle A Acob,Peizhu Guan,Andrea Blas,Ching Man Wai,Christine M Ackerman,Yan Ren,Chao Liu,Jianmei Wang,Jianping Wang,Jong-Kuk Na,Eugene V Shakirov,Brian Haas,Jyothi Thimmapuram,David Nelson,Xiyin Wang,John E Bowers,Andrea R Gschwend,Arthur L Delcher,Ratnesh Singh,Jon Y Suzuki,Savarni Tripathi,Kabi Neupane,Hairong Wei,Beth Irikura,Maya Paidi,Ning Jiang,Wenli Zhang,Gernot Presting,Aaron Windsor,Rafael Navajas-Pérez,Manuel J Torres,F Alex Feltus,Brad Porter,Yingjun Li,A Max Burroughs,Ming-Cheng Luo,Lei Liu,David A Christopher,Stephen M Mount,Paul H Moore,Tak Sugimura,Jiming Jiang,Mary A Schuler,Vikki Friedman,Thomas Mitchell-Olds,Dorothy E Shippen,Claude W dePamphilis,Jeffrey D Palmer,Michael Freeling,Andrew H Paterson,Dennis Gonsalves,Lei Wang,Maqsudul Alam Papaya, a fruit crop cultivated in tropical and subtropical regions, is known for its nutritional benefits and medicinal applications. Here we report a 3x draft genome sequence of 'SunUp' papaya, the first commercial virus-resistant transgenic fruit tree to be sequenced. The papaya genome is three ... ( view more )times the size of the Arabidopsis genome, but contains fewer genes, including significantly fewer disease-resistance gene analogues. Comparison of the five sequenced genomes suggests a minimal angiosperm gene set of 13,311. A lack of recent genome duplication, atypical of other angiosperm genomes sequenced so far, may account for the smaller papaya gene number in most functional groups. Nonetheless, striking amplifications in gene number within particular functional groups suggest roles in the evolution of tree-like habit, deposition and remobilization of starch reserves, attraction of seed dispersal agents, and adaptation to tropical daylengths. Transgenesis at three locations is closely associated with chloroplast insertions into the nuclear genome, and with topoisomerase I recognition sites. Papaya offers numerous advantages as a system for fruit-tree functional genomics, and this draft genome sequence provides the foundation for revealing the basis of Carica's distinguishing morpho-physiological, medicinal and nutritional properties. ( view less ) Xiao-Jian Wu,Jian-Ping Wang,Lei Wang,Xiao-Sheng He,Yi-Feng Zou,Lei Lian,Long-Juan Zhang,Ping Lan BACKGROUND: Total mesorectal excision (TME) has increased the rate of sphincter-preservation (SP) for more patients with low-lying rectal cancer. Here, we analyze the change of sphincter preserving rates in lower rectal cancer and their related factors. METHODS: We reviewed retrospectively the medi... ( view more )cal records of 316 patients with lower rectal cancers, 1 to 5 cm from the anorectal line, who had surgical resections from August 1994 to November 2005. The 12-year span was divided into 2 periods: period I (August 1994-December 1998) and period II (January 1999-November 2005), based on the date (January 1999) when standard total mesorectal excision (TME) was introduced. The patients were divided into two groups based on the operation: abdominoperineal resection (APR) or SP surgery. SP rates, leakage and other clinico-pathological characteristics were compared between the two time periods and between the two different groups. RESULTS: The SP rate increased significantly over the 12 years, from 44.9% in period I to 76.2% in period II (P = 0.000). The factors significantly influencing SP included the distance of the tumor from the anorectal line, gender, time period, circumference of intramural spread and histological differentiation (P < 0.05). Significant differences were detected between the two time periods in gender, blood transfusion volume and Dukes' stage (P < 0.05). The leakage rate was 2.7% in period I and 1.3% in period II (P > 0.05). CONCLUSIONS: Over the 12-year period of the study the SP rate in rectal cancers 1 - 5 cm from the anorectal line has increased significantly while the blood transfusion volume has decreased due to the introduction of TME. However, TME had no effect on operating time and leakage rates. ( view less ) Qian Lei,Lei Chen Heparin-induced thrombocytopenia (HIT) is an antibody-mediated complication of heparin treatment that can lead to thrombosis and thromboembolism. HIT is mainly caused by immunoglobulin G (IgG) class among anti-heparin/platelet factor 4 antibodies that bind to epitopes on platelet factor 4 (PF4) rel... ( view more )eased from activated platelets that developed when it forms complexes with heparin. Platelet aggregation and hypercoagulation status result from this process. Besides, the reactions between antibodies and vascular endothelial cells and monocytes are involved in HIT. Laboratory detection of anti-heparin/platelet factor 4 antibodies after heparin administration may help diagnose HIT early. Tests for detecting antibodies to the heparin/PF4 complex can be classified into functional platelet assays (which rely on the demonstration of platelet activation) and immunoassays (which detect the presence of an antibody without regard for its functional ability). But there is no simple and effective test available currently. In this article the anti-heparin/platelet factor 4 antibodies, pathogenesis of HIT, clinical laboratory assays and immunoassays are reviewed. ( view less ) Lei Zhang,Lei Dong,Haipeng Dou,Wangbao Yin,Suotang JiaLaser-induced breakdown spectroscopy has been used to measure the organic oxygen content in pulverized anthracite coal under atmospheric conditions. Special spectral processing including the optimal O(I) emission-line selection by comparing the spectral correlation coefficients with the N(I) line, ... ( view more )internal normalization with the N(I) line, and temperature correction are proposed and employed to satisfy the multi-line analysis method and yield the most accurate quantitative results. The calibration method for determining the organic oxygen content of coal is presented, with an accuracy of 1.15-1.37% and an average relative error of 19.39% being evaluated through an experiment performed on six anthracite coal samples. The relative measurement error distribution has also been studied. ( view less ) Bohua Li,Shu Shi,Weizhu Qian,Lei Zhao,Dapeng Zhang,Sheng Hou,Lei Zheng,Jianxin Dai,Jian Zhao,Hao Wang,Yajun Guo Despite the effectiveness of the anti-CD20 monoclonal antibody (mAb) Rituximab (C2B8) in the treatment of B-cell lymphoma, its efficacy remains variable and often modest. It seems likely that a combination of multiple mechanisms, such as complement-dependent cytotoxicity (CDC) and apoptotic signali... ( view more )ng, underlies the therapeutic success of anti-CD20 mAbs. Unfortunately, all the current anti-CD20 mAbs effective in CDC are relatively inactive in signaling cell death and vice versa. In this study, we developed two genetically engineered tetravalent antibodies (TetraMcAb) respectively derived from the anti-CD20 mAbs C2B8 and 2F2. TetraMcAbs, with a molecular mass only 25 kDa higher than native divalent antibodies (DiMcAb), were shown not only to be as effective in mediating CDC and antibody-dependent cellular cytotoxicity against B-lymphoma cells as DiMcAbs but also to have antiproliferative and apoptosis-inducing activity markedly superior to that of DiMcAbs. Interestingly, whereas 2F2 and C2B8 were equally effective in inducing cell growth arrest and apoptosis, the functions of their tetravalent versions, 2F2(ScFvHL)(4)-Fc and C2B8(ScFvHL)(4)-Fc, were significantly different. 2F2(ScFvHL)(4)-Fc exhibited exceptionally more potent antiproliferative and apoptosis-inducing activity than that of C2B8(ScFvHL)(4)-Fc. Immunotherapeutic studies further showed that 2F2(ScFvHL)(4)-Fc was far more effective in prolonging the survival of severe combined immunodeficient mice bearing systemic Daudi or Raji tumors than C2B8, 2F2, and C2B8(ScFvHL)(4)-Fc, suggesting that it might be a promising therapeutic agent for B-cell lymphoma. ( view less ) Lei Yu,Xiaolin Pei,Ting Lei,Yuhua Wang,Yan Feng Genome shuffling is a powerful strategy for rapid engineering of microbial strains for desirable industrial phenotypes. Here we applied the genome shuffling to improve the glucose tolerance of Lactobacillus rhamnosus ATCC 11443 while simultaneously enhancing the L-lactic acid production. The starti... ( view more )ng population was generated by ultraviolet irradiation and nitrosoguanidine mutagenesis and then subjected for the recursive protoplast fusion. The positive colonies from library created by fusing the inactivated protoplasts were more likely to be screened on plates containing different concentrations of high glucose and 2% CaCO(3). Characterization of all mutants and wild-type strain in the shake flask indicated the compatibility of two optimal phenotypes of glucose tolerance and lactic acid enhancement. The lactic acid production, cell growth and glucose consumption of the best performing strain from the second round genome shuffled populations were 71.4%, 44.9% and 62.2% higher than those of the wild type at the initial glucose concentration of 150 g/l in the 16l bioreactor. Furthermore, the higher lactic acid concentrations were obtained when the initial glucose concentrations increased to 160 and 200 g/l in batch fermentation. ( view less ) Weiwei Gai,Wei Zou,Lei Lei,Junyi Luo,Haobo Tu,Yan Zhang,Kai Wang,Po Tien,Huimin Yan Severe acute respiratory syndrome (SARS) is a deadly and highly infectious disease caused by SARS Coronavirus (SARS-CoV). Inactivated SARS-CoV has been explored as a vaccine against SARS-CoV; however, current knowledge of inactivated SARS-CoV vaccine is quite limited. We attempted to investigate th... ( view more )e effects of different immunization protocols and adjuvant on the antibody responses to inactivated SARS-CoV vaccine. With an intraperitoneal (IP) immunization protocol, inactivated SARS-CoV alone induced significant amounts of SARS-CoV-specific IgG antibodies in sera and a small quantity of SARS-CoV-specific IgA antibodies in the genital tract and feces, but failed to induce any detectable SARS-CoV-specific IgA antibodies in sera, saliva, lung, and intestine, and the addition of CpG ODN 2006 had only a marginal effect on antibody production. In contrast, with an intranasal (IN) immunization protocol, inactivated SARS-CoV alone failed to induce any detectable SARS-CoV-specific IgA antibodies in sera, saliva, lung, and intestine, except for a small quantity of IgA antibodies in fecal extracts and the genital tract, along with IgG antibodies in sera, but when given with adjuvant CpG ODN 2006, inactivated SARS-CoV induced significant amounts of SARS-CoV-specific IgG antibodies in sera, and a detectable amount of SARS-CoV-specific IgA antibodies in sera and all tested mucosal secretions and tissues (i.e., saliva, the genital tract, fecal extract, lung, and intestine). On a neutralization assay, neutralizing activity with the IP immunization protocol was detected in sera and mucosal secretions (from the saliva and genital tract), but sera from the IN protocol failed to show any neutralizing activity. Our study demonstrated that inactivated SARS-CoV vaccine is promising, and our data provide a sound foundation for the development of an effective inactivated SARS-CoV vaccine. ( view less ) Xiu-Qing Feng,Ying-Wei Lin,Ya-Jun Chen,Shu-Qi Zhong,Xiao-Fei Yan,Jian-Jiang Dong,Lei Lei In this study we detected dynamic changes and function of beta-tubulin, a subtype of microtubule, during the first cleavage period in mouse parthenogenetic and in vitro fertilized embryos. Firstly, we compared the developmental potential of in vitro fertilized, parthenogenetic, and in vivo fertiliz... ( view more )ed embryos in culture. Then, the dynamic changes of beta-tubulin and nucleus in parthenogenetic and in vitro fertilized preimplantation embryos were detected by immunofluorescence and confocal microscopy to analyze the role of microtubules in meiotic division and embryonic development. The results indicated that the development rate of in vivo fertilized embryos was significantly higher than that of in vitro fertilized or parthenogenetic embryos (P<0.05). However, there was no significant difference in developmental potential between in vitro fertilized and parthenogenetic embryos. During in vitro fertilization, oocyte was activated when sperm entered it. Oocyte resumed the second meiotic division. Condensed maternal chromosomes aligning at the equator of the spindle were pulled to the spindle poles by kinetochore microtubules in anaphase. Furthermore, in telophase, there were microtubules between the two sets of decondensed maternal chromosomes. One set formed the second polar body (Pb(2)), which was extruded to the perivitelline space. The other set formed female pronucleus. Meanwhile, 5-8 h after fertilization, sperm chromatin condensed and decondensed to form male pronucleus. Microtubule composed mesosome and cytaster remodeling around male and female pronuclei to form long microtubules, which pull the pronuclei to get close. During 4-6 h parthenogenetic activation, SrCl2 activated oocytes to resume meiosis. As a consequence, sister chromatids were pulled to spindle poles. Cytochalasin B, which was applied in the medium, inhibited the extrusion of Pb(2). Two haploid pronuclei in the cytoplasm were connected by microtubules. Compared with that in in vitro fertilization, oocyte is easier to be activated in parthenogenetic activation. Chemical activation is more efficient than sperm penetration in in vitro fertilization as indicated by earlier and better remodeling of the microtubules. ( view less )
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