Mechanism of the reconstruction of aqueous outflow drainage

Glaucoma is the leading cause of irreversible blindness worldwide. The reconstruction of aqueous outflow drainage (RAOD) has recently been proposed to aid in restoring aqueous outflow drainage in primary open-angle glaucoma. However, the mechanism of RAOD remains to be fully understood. Based on literature review and research studies, the potential mechanisms of RAOD are the following: (i) Circumferential dilation of the Schlemm’s canal (SC) and surrounding collector channels. (ii) Instant formation of microcracks through RAOD procedures. (iii) Formation of more pores, and local detachment between the SC endothelium (SCE) and basement membrane. (iv) Activation of stem cells by constant mechanical stress caused by the tensional suture placed at the anterior part of the SC. (v) Reversal of trabecular meshwork (TM) herniation. (vi) Mobilization of the reserve of the aqueous drainage. (vii) Change of SCE phenotype. (viii) Mechanosensing and mechanotransducing of TM.     

Three-dimensional structure and function study on the active region in the extracellular ligand-binding domain of human IL-6 receptor

In this study the three-dimensional (3-D) model of the ligand-binding domain (V106-P322) of human interleukin-6 receptor (hlL-6 R) was constructed by computer-guided ho-mology modeling technique using the crystal structure of the ligand-binding domain (K52-L251) of human growth hormone receptor (hGHR) as templet. Furthermore, the active binding region of the 3-D model of hlL-6R with the ligand (hlL-6) was predicted. In light of the structural characteristics of the active region, a hydrophobic pocket shielded by two hydrophilic residues (E115 and E505) of the region was identified by a combination of molecular modelling and the site-directed or double-site mutation of the twelve crucial residues in the ligand-binding domain of hIL-6R (V106-P322). We observed and analyzed the effects of these mutants on the spatial conformation of the pocket-like region of hlL-6 R. The results indicated that any site-directed mutation of the five Cys residues (four conservative Cys residues: Cyst 21, Cys132, Cys165, Cys1     

Alpha-latrotoxin Triggers Extracellular Ca^2＋-dependent Exocytosis and Sensitizes Fusion Machinery in Endocrine Cells

α-Latrotoxin from the venom of black widow spider induces and augments neurotransmitterand hormone release by way of extracellular Ca~(2 ) influx and cellular signal transduction pathways.By usingwhole cell current and capacitance recording,the photolysis of card Ca~(2 ),and Ca~(2 ) microfluorometry andamperometry,we investigated the stimulating effect and mechá(?)ism of α-latrotoxin on exocytosis in ratpancreatic β cells,LβT2 cells and latrophilin plasmid-transfected INS-1 cells.Our data indicated that:(1)α-latrotoxin increased cytosolic Ca~(2 ) concentration through the formation of cation-permitting pores and sub-sequent Ca~(2 ) influx with the presence of extracellular Ca~(2 );(2)α-latrotoxin stimulated exocytosis in normalbath solution and its stimulating effect on secretion was eradicated in Ca~(2 )-free bath solution; and (3)α-latrotoxin sensitized the molecular machinery of fusion through activation of protein kinase C and increasedthe response of cells to Ca~(2 ) photolysed by a flash of ultraviolet light.In summary,α-latrotoxin inducedexocytosis by way of Ca~(2 ) influx and accelerated vesicle fusion by the sensitization of fusion machinery.     

Developmental changes in functional expression andβ-adrenergic regulation of I_f in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneous rate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart, therefore we studied developmental changes in functional expression and β-adrenergic regulation of Iy in embryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus) ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytes and even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating that these cells of the EDS embryonic heart have some properties of pacemaker cells.β-adrenergic agonist isoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that the β-adrenergic regulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase) and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase t     

《真菌学报》编辑委员会EDITORIAL BOARD

<正> 主编 Editor-in-Chief:王云章 WANG Yun-chang (Institute of Microbiology, Academia Sinica, Beijing)副主编 Associate editors-in-Chief:相望年 XIANG Wangnian (Institute of Microbiology, Academia Sinica, Beijing)魏江春 WEI Jiang-chun (Institute of Microbiology, Academia Sinica, Beijing)余永年 YU Yong-nian (Institute of Microbiology, Academia Sinica, Beijing)编委 Members of the Editorial Board:王鸣歧 WANG Ming-qi (Fudan University, Shanghai)齐祖詷 QI Zu-tong (Institute of Microbiology, Academia Sinica, Beijing)     

黄瓜花叶病毒衣壳蛋白基因转化辣椒研究

The plant expression vector of the coat protein(CP) gene of cucumber mosaic virus (CMV) BS strain was used to transform three kinds of pepper (Capsicum annuum) tissues (cotyledon, stem and root) by agrobacterium-mediated co-cultivation. 53%-68.4% of the total tissues (639) can be induced to be calli, but only cotyledon calli can be further regenerated to form shoots (regenerated efficiency 39.7%). 70%(42/60) of the putative transformed plants were confirmed to have CP gene in their genomes by Southern blot. The mRNAs and the CP were respectively found in 80% of transgenic plants by Northern blot and DAS-ELISA. 24 of the transgenic plants expressing CP gene of BS strain showed three kinds of resistant level (severe symptom, delay of symptomatic development, no symptom) to infection of CMV-BS and of CMV-P. However, there was distinctly higher resistance to inoculation of CMV-BS than that with CMV-P in these transgenic plants.     

Gene cloning, expression and purification of human mitochondrial tRNA~(Leu(UUR)) and its mutant

Genes of human mitochondrial tRNALeu(UUR) (mtRNALeu(UUR)) and its mutant (mtRNALeu(M)) were synthesized and inserted into the plasmid pGEM-9Zf(-) respectively. E.coli JM 109 was transformed by the recombinant plasmids containing the target genes. The mtRNALeu(UUR) and mtRNALeu(M) were expressed up to 19.10% and 17.76% of total small RNA respectively. They were purified to 54% homogeneity by DEAE-sepharose-CL4B column chromatography and finally repu-rified by 15% PAGE/urea. Their kinetic parameters for E.coli LeuRS were measured. The results showed that the value of kcal/Km of mtRNALeu(M) was about one fifth of that of mtRNALeu(UUR) and indicated the leucine acceptability of mtRNALeu(M) was much lower than that of mtRNALeu(UUR).     

The Ninth Editorial Board of Acta Microbiologica Sinica

CONSULTANT Jilun Li,Academician(College of Biology,Chinese Agricultural University,Beijing100094 ,China)EDITOR-IN-CHIEF Huarong Tan,Professor(Institute of Microbiology,Chinese Academy of Sciences,Beijing100101 ,China)VICE-EDITOR-IN-CHIEF Xiuzhu Dong,Professor(Institute of Microbiology,Chinese Academy of Sciences,Beijing100101 ,China)     

A new genus and species of Eulophidae (Hymenoptera:Chalcidoidea) parasitizing Hyphantria cunea(Drury) (Lepidoptera:Arctiidae) in China

In the spring of 1985, a species of chaicid wasps was reared from the pupae of fall webworm, Hyphantria cunea (Drury) (Lepidoptera:Arctiidae) during myinvestigation on the parasitoids of this introduced harmful insect pest inShaanxi Province, China. With high parasitic ratio (over 80 percent) and a largenumber of individuals (over 200 were usually reared from a single pupa of the     

Testing for the presence of magnetite in the upper-beak skin of homing pigeons

We carried out magnetic and nonmagnetic experiments on fresh, upper-beak skin tissue samples isolated from six pairs of homing pigeons to test whether the tissue contains magnetite particles. Results of (1) room-temperature isothermal remanent magnetization (IRM) acquisition and alternating field (AF) demagnetization, (2) low-temperature demagnetization of saturation IRM acquired at 5 K in a field of 5 tesla (T) (SIRM_{5 K}) after zero-field cooled (ZFC) and field cooled (FC) treatments, and (3) cycling of the saturation IRM acquired at 300 K in a field of 5 T (SIRM_{300 K}) between 5 and 300 K, indicate the presence of magnetite in the measured samples. A significant loss of SIRM_{5 K} below 20 K suggests the dominance of superparamagnetic (SPM) particles. The SIRM acquisition capacity of the female pigeon is stronger than that of the male pigeon in all four measured pairs, suggesting for the first time that the magnetite concentration is probably sex dependent. Light microscopic observation on the histological sections stained with Prussian Blue detected the presence of some tiny, dotted, dark-blue staining Fe³⁺ aggregates (size 1–4 μm) located directly beneath the subcutis within strands of connective tissue, nearby the rim of the regions full of red nuclei. The results of this study support the idea that homing pigeons may have a magnetite-based receptor, which potentially could be used for sensing the Earth’s magnetic field during navigation.     

The Ninth Editorial Board of Acta Microbiologica Sinica

CONSULTANT Jilun Li,Academician(College of Biology,Chinese Agricultural University,Beijing 100094 ,China)EDITOR-IN-CHIEF Huarong Tan,Professor(Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101 ,China)VICE-EDITOR-IN-CHIEF Xiuzhu Dong,Professor(Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101 ,China)     

Rearrangement of Actin Cytoskeleton by Zika Virus Infection Facilitates Blood–Testis Barrier Hyperpermeability

In recent years,various serious diseases caused by Zika virus (ZIKV) have made it impossible to be ignored.Confirmed existence of ZIKV in semen and sexually transmission of ZIKV suggested that it can break the blood–testis barrier (BTB),or Sertoli cell barrier (SCB).However,little is known about the underlying mechanism.In this study,interaction between actin,an important component of the SCB,and ZIKV envelope (E) protein domain Ⅲ (EDⅢ) was inferred from coimmunoprecipitation (Co-IP) liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis.Confocal microscopy confirmed the role of actin filaments (F-actin) in ZIKV infection,during which part of the stress fibers,the bundles that constituted by paralleled actin filaments,were disrupted and presented in the cell periphery.Colocalization of E and reorganized actin filaments in the cell periphery of transfected Sertoli cells suggests a participation of ZIKV E protein in ZIKV-induced F-actin rearrangement.Perturbation of F-actin by cytochalasin D (CytoD) or Jasplakinolide (Jas)enhanced the infection of ZIKV.More importantly,the transepithelial electrical resistance (TEER) of an in vitro mouse SCB (mSCB) model declined with the progression of ZIKV infection or overexpression of E protein.Co-IP and confocal microscopy analyses revealed that the interaction between F-actin and tight junction protein ZO-1 was reduced after ZIKV infection or E protein overexpression,highlighting the role of E protein in ZIKV-induced disruption of the BTB.We conclude that the interaction between ZIKV E and F-actin leads to the reorganization of F-actin network,thereby compromising BTB integrity.     

人巨细胞病毒形态转化区的鉴定

DNA of human cytomegalovirus (HCMV) contains three transforming fragments, which have been mapped in the long unique region of the viral genome(see Fig.1). A minimal region of 558 base pairs(bp) (pCM4127) was localized in the XbaI-Hindlll fragment of HCMV strain AD169 (mapunit 0.123—0.140)and designated morphological transforming region Ⅰ (mtrⅠ). MtrⅠwas reported to cause one-step focal transformation of primary Wistar     

Developmental changes in functional expression and β-adrenergic regulation of If in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneousrate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart,therefore we studied developmental changes in functional expression and β-adrenergic regulation of If inembryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus)ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytesand even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating thatthese cells of the EDS embryonic heart have some properties of pacemaker cells. β-adrenergic agonistisoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that theβ-adrenergicregulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase)and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of If in EDS cells,indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore,the results demonstrate that If is modulated by phosphorylation via cAMP dependent PKA both in EDSand LDS cells.     

Developmental changes in functional expression and β-adrenergic regula-tion of If in the heart of mouse embryo

The hyperpolarization-activated current (If) plays an important role in determining the spontaneous rate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart,therefore we studied developmental changes in functional expression and β-adrenergic regulation of If in embryonic mouse heart. The expression of If is high in early developmental stage (EDS) (10.5 d after coitus) ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytes and even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating that these cells of the EDS embryonic heart have some properties of pacemaker cells, β-adrenergic agonist isoproterenol (ISO) stimulates If in LDS but not in EDS cardiomyocytes, indicating that the β-adrenergic regulation of If is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase) and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of If in EDS cells,indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore,the results demonstrate that If is modulated by phosphorylation via cAMP dependent PKA both in EDS and LDS cells.     

The 2018 Nobel Prize in Chemistry: Engineering proteins (enzymes/peptide/antibodies) towards desired properties via the construction of random libraries

<正>The 2018 Nobel Prize in Chemistry was awarded one half to Dr. Frances Arnold (of California Institute of Technology,USA) for her work on "directed evolution of enzymes", the other half to Dr. George Smith (of the University of Missouri, USA) and Sir Gregory Winter (of MRC Laboratory of Molecular Biology, UK) for their work on "phage display of peptides and antibodies".     

Purification and Characterization of a New Ribosome Inactivating Protein from Cinchonaglycoside C-treated Tobacco Leaves

A new ribosome-inactivating protein(RIP)with a molecular weight of 31 kDa induced by Cinchonaglycoside C(1)designatedCLP31,was isolated from tobacco leaves.Analysis of this protein sequence indicated that it belongs to the RIP family and itwas distinct from the other plant RIPs reported previously at its N-terminal amino acid sequence.CIP31 can directly impairsynthesis of coat protein(CP)of tobacco mosaic virus(TMV),which resulted in inhibition of TMV long distance movementand multiplication in tobacco plants at concentrations of ng/mL.Furthermore,no toxicity was shown to the growth andfertility of the plants.CIP31 was synthesized only in the presence of Cinchonaglycoside C(1)and was independent of thesalicylic acid(SA)signal pathway.We provided evidence for the SA-independent biological induction of resistance.     

Energy Transfer among Chlorophylls in Trimeric Light-harvesting ComplexⅡ of Bryopsis corticulans

A study on energy transfer among chlorophylls(Chls)in the trimeric unit of the major light-harvesting complex Ⅱ(LHC Ⅱ)from Bryopsis corriculan,was carried out using time-correlated singlephoton counting.In the chlorophyll Q region of LHC Ⅱ,six molecules characterized as Chlb_(628),Chlb_(646),Chlb_(652)~(654,657),Chla_(664)~(666),Chla_(674)~(677.680)and Chla_(682)~(683) were discriminated according to their absorption spectrumand fluorescence emission spectrum.Then,excited by pulsed light of 628 nm,fluorescence kinetics spectrain the chlorophyll Q region were measured.In accordance with the principles of fluorescence kinetics,thesekinetics data were analyzed with a multi-exponential model.Time constants on energy transfer were obtained.An overwhelming percentage of energy transfer among chlorophylls undergoes a process longer than 97picoseconds(ps),which shows that,before transferring energy to another Chl,the excited Chl might convertenergy to vibrations of a lower state with different multiplicity(intersystem crossing).Energy transfer at thelevel of approximately 10 ps was also obtained,which was interpreted as the excited Chls may go throughinternal conversion before transferring energy to another Chl.Although with a higher standard deviation,timeconstants at the femtosecond level can not be entirely excluded,which can be attributed to the ultrafastprocess of direct energy transfer.Owing to the arrangement and direction of the dipole moment of Chls inLHC Ⅱ,the probability of these processes is different.The fluorescence lifetimes of Chlb_(652)~(654,657),Chla_(664)~(666),Chla_(674)~(677.680)and Chla_(682)~(683)were determined to be 1.44ns,1.43 ns,636 ps and 713 ps,respectively.Thepercentages of energy dissipation in the pathway of fluorescence emission were no more than 40% in thetrimeric unit of LHC Ⅱ.These results are important for a better understanding of the relationship between thestructure and function of LHC Ⅱ.     

Skeletal muscle-specific expression of human blood coagulation factor Ⅸ rescues factor Ⅸ deficiency mouse by AAV-mediated gene transfer

The efficacy of recombinant adeno-associated virus (AAV) vector to deliver and express human blood clotting factor DC (hFIX) gene in skeletal muscle of coagulation factor IX deficiency mouse strain (FactorIX-knockout) is e-valuated. The muscle creatine kinase enhancer (MCK) and βactin promoter ((3A) were used to drive the hFIX minigene (hFIXml), which was flanked by AAV inverted terminal repeats (ITRs). Following intramuscular injection of high liter (2.5 x 1011 vector genomes/mL) of AAV, increased hFIX expression (256 ng/mL of plasma) was achieved. The time course of hFIX expression demonstrated that the expression level gradually increased over a period of two weeks before anti-hFIX antibodies developed in mouse circulating plasma. Those results provided a promising evidence that rAAV-me-diated gene transfer and skeletal muscle-specific expression of hFIX is a feasible strategy for treating patients for hemophilia B.