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Factors Influencing Agrobacterium-mediated Transformation of Foxtail Millet (Setaria italica)

Yinghui LIU ; Jingjuan YU ; Guangming AO ; Qian ZHAO

Chinese Journal of Biochemistry and Molecular Biology.2007;23(7):531-536.

An efficient and repeatable approach in transforming the foxtail millet (Setaria italica ) using Agrobacterium LBA4404 horboring the plasmid pBI121 was established. Factors affecting transformation efficiency were investigated including the genotype, explants, the density of bacteria, the duration for inoculation and co-cultivation, and the concentration of acetosyringone in the medium. The maximum transformation conditions were: the callus induced from inflorescence was used as explant; the duration for inoculation with Agrobacterium at low cell density was 30-40 min; for the co-cultivation , the suitable concentration of acetosyringone in the medium was 0.1 mmol/L, and the duration was 2 days.

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Molecular Characterization and Functional Prediction of a Novel Leaf SAG Encoding a CBS-domain-containing Protein from Coleus blumei

Qinlong ZHU ; Mingyang LI ; Guangde LIU ; Yandong LI ; Shunzhao SUI ; Tieying GUO

Chinese Journal of Biochemistry and Molecular Biology.2007;23(4):262-270.

Leaf senescence is considered as one of important factors to decrease ornamental values of foliage plants. In the attempt to study and understand the molecular mechanism of leaf senescence, a senescent leaf cDNA library of Coleus blumei was constructed and a small EST library was obtained. According to the sequence of an EST fragment with a cystathionine beta synthase (CBS) domain, a novel leaf senescenceassociated gene (SAG) full-length cDNA encoding a CBS-domain-containing protein, denoted Cbcbs, was rapidly cloned using a strategy of RACE combined with cDNA library. The full length of the Cbcbs gene was 859 bp long (accession No. EF076754) and contained a 609 bp open reading frame (ORF) encoding a 202amino acid protein. One stop codon (TAA) was found in 5' UTR and one possible polyadenylation signal,AATAAA, and a pentanucleotide motif, ATTTA, were found in 3' UTR. The CbCBS contained a predicted mitochondrial targeting peptide in the N-terminal region, two conserved and intact CBS domains, four casein kinase Ⅱ (CK Ⅱ) phosphorylation sites, three protein kinase c (PKC) phosphorylation sites and one tyrosine sulfation (TS) site. Sequence comparisons and phylogenetic analysis showed that CbCBS was a novel senescence or stress-associated protein. The prediction analysis of secondary structure and three dimensional structure of CbCBS suggested that the chief function of the protein was decided by the CBS domain pair. The expression pattern of Cbcbs in leaves was analyzed by RT-PCR. It was demonstrated that Cbcbs gene was a senescence-associated gene (SAG) and expressed in all leaf stages, young stage (Y) being the lowest and terminal senescence stage (S3) being the highest, and was upregulated along with the leaf senescence.Function analysis showed that the mature CbCBS maybe acts as a sensor of cellular energy status and directly or indirectly regulates cellular energy levels to increase ATP content in mitochondria during periods of metabolic stress of senescent leaves.

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Identification of the Kiwifruit Germplasms in Jiangxi Province by AFLP and Its Classification Significance

Hua CHEN ; Ganjun YI ; Xiaobiao XU

Chinese Journal of Biochemistry and Molecular Biology.2007;23(2):122-129.

The kiwifruit germplasms in Jiangxi province were identified by amplified fragment length polymorphism(AFLP)markers．Four primer pairs that had been selected from 64 ones had detected a total of 190 bands among 31 germplasms of kiwifruit,one hundred and seventy nine bands that representing 94．2％of total bands were polymorphic．The identification rates of 31 germplasms were up to 100％．The result suggests that applying AFLP markers to analyze the kiwifruit germplasms is feasible．Then Clustering analysis the AFLP resuit by UPGMA．The dendrogram indicated that the Dice similarity coefficients of 31 germplasms of kiwifruit ranged from 0．50 to 0．85,it suggested that their genetic relationships weren't near．Thirty one germplasms could divide into four groups according to the Dice similarity coefficient 0．56．Sect．Leiocarpae and Sect．Maculatae clustered into one group；A．Melliana of Sect．Strigosae was one group；A．Chinensis var. Rufopulpa,A．Eriantha,A．Fulvicoma var．Lenata,A．Styracifolia and A．Jiangxiensis in Sect．Stellatae clustered into the same group；but the two germplasms of Sect．Stellatae,A．Deliciosa and A．Chinensis clustered into the other group．Interestingly,the‘Ganmi-5’was in the same rank with A．Chinensis from the dendrogram,but it was the variety of A．Chinensis according to the traditional classification．It is necessary that the‘Ganmi-5’should be further researched in classification．The genetic relationships among the kiwifruit germplasms in Jiangxi province were identified and characterized by themolecular method,which was consistent with the traditional classification in certain degree and provided new evidences for the classification of the kiwifruit germplasms．

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Recombinant Human PLD2(rhPLD2)May Significantly Inhibit Expression of GPI-PLD of Guinea Pigs of Chronic Asthma in vivo

Ling ZHU ; Chuanxing YU ; Weibin ZOU ; Xiaoli HE ; Junjin LIN

Chinese Journal of Biochemistry and Molecular Biology.2007;23(2):116-121.

The effect of recombinant human phospholipase D2(rhPLD2)in vivo was investigated on the secretion of serum glycosyl phosphatidylinositol-specific phospholipase D(GPI-PLD)in guinea pigs of chronic asthma．Ater treating the guinea pigs attacked by chronic asthma with rhPLD2,the GPI-PLD activity detection was canrried out by phase separation of human placental alkaline phosphatase in Triton X-114．Compared with the healthy guinea pigs(NS group),the serum GPI-PLD in the guinea pigs of chronic asthma are much higher than that of control groups,P≤0．01．Our results showed that rhPLD2 could significantly reduce the secretion of GPI-PLD when the guinea pigs were attacked by chronic asthma．

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Old Enzyme with New Interest:Review of Newly Understood NEP Functions and Its Potential Renglators

Yue HONG ; J.turner ANTONY

Chinese Journal of Biochemistry and Molecular Biology.2007;23(2):85-92.

Neprilysin(NEP)is a type Ⅱ integral membrane glycoprotein of the M13 zinc metalloprotease family．As a neuropeptide degrading enzyme,NEP has been discovered to possess an increasing amount of organ-specific functions from central nervous system to peripheral tissues since it was fimfly identified in 1974．For example,NEP has been shown to have an anti-tumour effect by inhibition of cell migration and proliferation while induction of an programmed cell death dependent of both its enzymatic activity and direct protein-protein interactions with key molecules involved in signal transduction pathways:NEP Was also implicated to have neuropmtective effect by preventing the accumulation of the neurotoxic amyloid β-peptide (Aβ)in brain．Through investigating the progression of various human diseases,impaired NEP expression and activity were found to occur frequently．Based on these findings,modulation of NEP levels in pathological cells is considered to be therapeutically applicable as a strategy to recover normal cell functions and thereafter relieve symptoms of diseases．Great research effort is being contributed to the study of regulatory mechanisms involved in expression and activity of this enzyme,and a number of encouraging results have already been achieved．Besides androgens,well-recognised regulators of NEP transcription in prostate,the female hormone oestrogen,aqueous extract of willow herb,components of green tea and neuropeptides bombesin,somatostatin as well as the intracellular domain of amyloid precursor protein were all shown to have a stimulatory effect on NEP expression and／or its activity．

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Prostate Cancer Bone Metastasis: Interaction Between Tumor Cells and Bone Microenvironment

Yi LU ; Zhi YAO ; Jian ZHANG

Chinese Journal of Biochemistry and Molecular Biology.2007;23(3):1-171.

Cancer metastasis is composed of a complex cascade that involves a variety of critical steps beginning with detachment from the primary tumor and ending with growth of tumor at a distant site, such as bone. The "seed-and-soil hypothesis" predicts that the bone microenvironment expresses factors through which attract a variety of cancer cells and promote the tumor development. The ending point of tumor development in bone is achieved through the bidirectional and dynamic interaction between tumor cells and the cells in their growth microenvironment. A variety of factors produced by the bone microenvironment, contribute to the pathogenesis of cancer skeletal metastasis. In this review, using prostate cancer (CaP) as an example, some of general mechanisms of cancer metastasis will be summarized. In addition, the current understanding of the interaction between tumor cells and the bone microenvironment will be addressed. Finally, the research directions in the near future will be suggested.

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Regulation of Cell Proliferation and Apoptosis by ERK Signaling Pathway in Airway Smooth Muscle Cells of Rats

Jing BAI ; Xiansheng LIU ; Yongjian XU ; Min XIE

Chinese Journal of Biochemistry and Molecular Biology.2007;23(1):38-44.

The regulative role of extracellular signal-regulated kinase(ERK)signaling pathway on proliferation and apoptosis in rats' airway smooth muscle cells(ASMCs)was investigated.Primary cultures of ASMCs were established and cells between passages 4 and 7 were used for experiments.ASMCs were U-eated with ERK activator epidermal growth factor(EGF)and inhibitor PD98059.The expressions of ERK mRNA and protein were detected by RT-PCR and immunofluorescence staining.Proliferation of ASMCs were detected by MTT eolorimetric assay and [3H] thymidine incorporation.Apoptosis of ASMCs were detected by Hoechst staining and Annexin-V FITC PI donble staining.The levels of ERK1/2,phosphorylated forms of ERK1/2(p-ERK1/2) and proeaspase-3 protein were detected by Western blotting.The expressions of ERK mRNA and ERK protein were obviously observed in ASMCs. Compared with control,the absorbanee(A490) value and DNA synthesis index of PD-treated ASMCs were significantly decreased(P＜0.05).The apoptotic index and the percentage of the early apoptotic cells were significantly increased(P＜0.05).The expressions of ERK1/2 and pERK1/2 protein were significantly down-regulated.The expressions of procaspase-3 protein was significantly increased.Compared with c,ontrol,the A490 value and DNA synthesis index were significantly increased(P＜0.05)in EGF-treated cells.Apoptotic index and the percentage of the early apoptotic cells were significantly decreased(P＜0.05).The levels of ERK1/2 and pERK1/2 protein were significantly increased,and the levels of procaspase-3 protein were significantly decreased, There were no significant differences between control and P+E group(P＞0.05).ERK signaling pathway may play an important role in regulating ASMCs proliferation and apoptosis and ERK regulating the apoptosis of ASMCs possibly relates to the expressions of procaspase-3 protein. The finding will help to understand the mechanisms of asthmatic ASMCs involved in abnormal proliferation.

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Column Guide

Chinese Journal of Biochemistry and Molecular Biology.2023;39(7):903-904. doi:10.13865/j.cnki.cjbmb.2023.06.1255

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The Mechanism of Nitric Oxide in Delaying Post-harvest Senescence of Agricultural Products

Li-Zhen LUO ; Yu-Zheng DENG ; Wei-Biao LIAO

Chinese Journal of Biochemistry and Molecular Biology.2023;39(7):933-943. doi:10.13865/j.cnki.cjbmb.2023.06.1128

Nitric oxide(NO) is essential for reducing post-harvest product senescence and storage losses. NO may slow down the senescence of agricultural products and improve their quality, suggestive of its great potential in the postharvest preservation of agricultural products. The development of NO as a post-harvest preservative is supported by the investigation of its role and mechanism in agricultural product ageing. In this review, the mechanisms of NO in the post-harvest senescence of agricultural products were explored. NO is produced mainly via the nitrate reductase(NR) and NO synthase-like(NOS) pathways. Usually, NO donors include sodium nitroprusside (SNP), S-nitroso-N-acetyl-DL-penicillamine (SNAP), S-nitrosoglutathione(GSNO) and NO gas fumigation. NO delays agricultural product senescence by regulating ROS metabolism, enhancing antioxidant activity, reducing respiration, activating cell wall metabolism, inhibiting ethylene synthesis, and promoting energy metabolism. Melatonin(MT), ethylene(ETH), hydrogen gas(H

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Regulation of H

Li-Ping ZHANG ; Lin-Lin ZHANG ; Hai-Yan CAO ; Wen-Qi LIU ; Zhi-Qiang LIU ; Dan-Mei LIU ; Zhu-Ping JIN ; Yan-Xi PEI ; Xue-Feng HAO

Chinese Journal of Biochemistry and Molecular Biology.2023;39(7):972-979. doi:10.13865/j.cnki.cjbmb.2023.06.1114

Hydrogen sulfide(H

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