Biomedical subjects

Jing Zhao

Publications and source records attributed to Jing Zhao.

At least 145 records · Page 8Linked to original sources

PubMed2004

[Establishment of Hela cell line inducibly expressing human active granzyme B].

AIM: To construct inducible expression vector for human granzyme B gene and express it in Hela cell line. METHODS: Human active granzyme B gene was obtained by PCR and cloned into the inducible expression vector pIND. The resulting expression vector, together with a helper plasmid pVgRXR, was stably transfected into Hela cells using Lipofectamine 2000. The transfected cells were selected in medium containing G418 and zeocin. The resistant cells were induced with ponasterone A, and the optimal concentration of ponasterone A and time of induction were determined by immunocytochemical staining. Then the effects of the expressed granzyme protein on Hela cells were detected by MTT colorimetry and cytoskeletal staining. RESULTS: The Hela cells that inducibly expressed human active granzyme B were obtained. Induction with 30 micromol/L ponasterone A for 5 days caused the strongest expression of granzyme B. The induced cells appeared as either multinucleate giant cells or pyknotic small cells, and their growth was inhibited. Further analysis demonstrated cytoskeletal abnormality of multinucleate giant cells. CONCLUSION: The establishment of inducible expression system for active granzyme B lays the foundation for further research on biological function of granzyme B.

Cell Proliferation↗

PubMed2004

[Organ culture for preservation of the cornea: human umbilical cord serum versus fetal bovine serum].

OBJECTIVE: To evaluate the changes of porcine corneal endothelium, the morphology, histology, ultrastructure, enzymes activity and metabolism of the cornea induced by organ culture with two different media containing fetal bovine serum (FBS) or human umbilical cord serum (HCS). METHODS: Fifty pairs of porcine corneas were preserved at 31 degrees C for 7, 14, 21, 28 days. One cornea of each pair was cultivated in medium I containing 10% FBS (group 1); the other one was stored in medium II containing 10% HCS (group 2). Thirteen fresh porcine corneas served as controls. All stored corneas were dehydrated for 24 hours. Twelve corneas from each group were evaluated each week, including the morphology, histology and enzyme histochemical staining of the cornea. Scanning electron microscopy was performed on one cornea from each group at 14 and 28 days and compared with the fresh cornea. pH value, glucose and lactate concentration of the culture media before and after culture were examined. Microbiological evaluation was also performed. RESULTS: Endothelium evaluation did not differ statistically between the two groups of porcine corneas. The morphological endothelium study showed some alterations such as pleomorphism. After 28 days of cultivation, the mean cell losses of endothelium were 10.98% and 10.85% in medium I and medium II stored corneas, respectively. There were no statistical differences of the histology, ultrastructure and enzymes activity of corneas between the two groups. The histological study showed corneal swelling and epithelial sloughing after preservation. Scanning electron microscopy showed an intact endothelial layer in all corneas. Enzyme histochemical staining showed vigorous enzyme activity in the corneal epithelium and endothelium. Enzyme activity in stroma decreased with preservation time. Corneas showed good glucose metabolism. Incidence of contamination was 6% for storage medium. CONCLUSIONS: The corneal endothelium can maintain a good viability for 4 weeks in these two organ culture media. HCS can replace FBS in the organ culture medium.

Animals↗

PubMed2004

Ion channel activities implicated in pathological pain.

Altered expression of voltage-gated sodium, calcium and potassium channels has been associated with neuropathic pain conditions. In addition, roles for the ligand-gated P2X3 and NMDA receptors, as well as pacemaker HCN channels have also been invoked in the pathogenesis of neuropathic pain. In this chapter, evidence of an important role for post-translational regulation of Nav1.9 in setting pain thresholds is presented. Despite the importance of tactile allodynia and mechanical hyperalgesia in chronic pain, we remain ignorant of the molecular nature of mechanosensors present in sensory neurons. A number of candidate mechanosensor genes, identified because of their structural similarity with mechanosensors in Caenorbabditis elegans and Drosophila melanogaster have been identified. Acid-sensing ion channels (ASICs) are structurally related to putative mechanosensors in C. elegans, whilst transient receptor potential channels (TRPs) have been implicated in mechanosensation in the Drosophila acoustic system. Evidence against a role for ASICs as primary transducers of mechanosensation is provided here, and recent evidence implicating TRP channels is reviewed. Finally, the use of sensory neuron-specific gene deletion approaches to unravel the significance of individual ion channels in the regulation of sensory neuron excitability and the induction of pain will be described.

Animals↗

PubMed2004

[Diagnosis and treatment of obstructive sleep apnea hypopnea syndrome in children].

OBJECTIVE: To discuss the methods of diagnosis and treatment of obstructive sleep apnea hypopnea syndrome (OSAHS) in children. METHODS: 285 patients diagnosed OSAHS by X-ray for lateral rhinopharyngeal view, fibrorhinopharyngescopy and overnight polysomnography ( PSG) were reviewed retrospectively the clinical symptoms and signs. The patients whose apnea hypopnea index(AHI) > or = 5/h, blood oxygen's saturation reducing over 0.03 are diagnosed as OSAHS. 255/285 cases were performed adenoidectomy and/or tonsillectomy and 9 cases were treated with continuous positive airway pressure (CPAP). RESULTS: The major symptoms of patients with OSAHS are snoring in sleep, breathing opening mouth, apnea, hearing loss. 281/285 cases were diagnosed as adenoid and (or) tonsil hypotrophy. Two hundred- fifty five cases received surgery, adenoidectomy and tonsillectomy in 205, only adenoidectomy in 47, simple tonsillectomy in 3. The clinical symptoms of 248 cases improved evidently after operations. Improved rate is 97.2%. In 1-3 months after surgery,PSG were checked in 105 patients again: AHI, the percentage of the sleep time for saturation monitored by pulse oximetry < 0.90, the longest time of apnea are improved than pre-operation. 9 cases using CPAP received good effects, 7 of 9 cases used CPAP were in pre- and /or post- surgery. Two cases were obesity hypopnea syndrome. CONCLUSIONS: Children with OSAHS has signification them-self; PSG is the major means for diagnosing. Surgery is main method for treating; CPAP can be as the method of treating for serious OSAHS in preoperative preparation and postoperative management.

Adenoidectomy↗

PubMed2004

[Growth inhibitory effects of recombinant granzyme B containing different N-terminal translocating peptides].

Translocating protein and translocating peptides have therapeutic potential against tumors by exposing the cytotoxic domains of toxic proteins to the cell cytosol. The aim of this study is to investigate the effect of N-terminally fused PE translocating peptides on granzyme B (GrBa) activity. PE II-GrBa fusion protein genes were constructed by replacing N-terminal signal and acidic dipeptide sequence of human granzyme B gene with two truncated translocating sequences of Pseudomonas exotoxin A (PE II aa 280-364/358) by recombinant PCR, and then cloned into pIND inducible expression vector. The resulting pIND-PE II-GrBa expression vectors were co-transfected with assistant plasmid pVgRXR into HeLa cells through lipofectamine, followed by selection on G418 and zeocin. The resistant cells were collected and induced with ponasterone A. Western blot analysis demonstrated that ponasterone A induction caused the expression of PE II-GrBa fusion proteins, and indirect immunofluorescence detected giant sized multinucleated cells, suggesting cytoskeletal and mitotic abnormalities as reported in our previous studies. Western blot, enzymatic activity assay and cell counting analysis indicated that two types of PE II-GrBa fusion proteins were capable of cleaving both endogenous and exogenous substrates of granzyme B, and inhibiting the growth of cells. The PE II (aa 280-358)-GrBa was shown to have higher serine protease activity and stronger growth inhibitory effect. Such inhibition was presumably associated with G2 arrest as determined by cell cycle analysis. These data prove that PE II-GrBa fusion proteins have cell inhibitory effect similar to GrBa, and that the shorter PE-derived peptide exerts less influence on GrBa activity. This study helps to optimize the construction of recombinant protein comprising translocating peptides and cytotoxic molecules for tumor cell killing.

ADP Ribose Transferases↗

PubMed2003

Distinct thermodynamics for the formation and cleavage of N-H bonds in aniline and ammonia. Directly-observed reductive elimination of ammonia from an isolated amido hydride complex.

The reactions of aryl and alkylamines with the (PCP)Ir fragment (PCP = 1,3-di-tert-butylphosphinobenzene) were studied to determine the reactivities and stabilities of amine and amido hydride complexes relative to C-H activation products. Reaction of aniline with the (PCP)Ir unit generated from (PCP)IrH2 and norbornene resulted in the N-H oxidative addition product (PhNH)(H)Ir(PCP) (1a). In contrast, reaction of this fragment with ammonia gave the ammonia complex (NH3)Ir(PCP) (2). The amido hydride complex that would be formed by oxidative addition of ammonia, (PCP)Ir(NH2)(H) (1b), was generated independently by deprotonation of the ammonia complex (NH3)Ir(H)(Cl)(PCP) (3) with KN(SiMe3)2 at low temperature. This amido hydride complex underwent reductive elimination at room temperature to form the ammonia complex 2. Addition of CO to anilide complex 1a gave (PCP)Ir(PhNH)(H)(CO) (4a). Addition of CNtBu to terminal amido complex 1b formed (PCP)Ir(NH2)(H)(CNtBu) (4b), the first structurally characterized iridium amido hydride. Complexes 4a and 4b underwent reductive elimination of aniline and ammonia; parent amido complex 4b reacted faster than anilide 4a. These observations suggest distinct thermodynamics for the formation and cleavage of N-H bonds in aniline and ammonia. Complexes 1a, 2, 4a, and 4b were characterized by single-crystal X-ray diffraction methods.

Journal Article↗

PubMed2003

[Prevalence of allergic respiratory disorders and skin prick test in Beijing urban and suburban children: a comparative study].

OBJECTIVE: To investigate the prevalence of allergic respiratory disorders and allergens in Beijing urban and suburban children. METHODS: Using the International Study of Asthma and Allergies in Childhood Phase III Protocol, a questionnaire survey was conducted among 7,077 junior middle school students, 3,531 males and 3,546 females, aged 13-14, of 11 urban schools and 10 suburban schools in Beijing by random cluster sampling. Skin prick test with 13 common allergens was conducted among 2,126 from the 7,077 students by random cluster sampling. RESULTS: The prevalence rates of anamnestic asthma, asthma in recent 12 months, asthma after exercise in recent 12 months, and dry cough during night in recent 12 months were 12.4%, 7.2%, 25.6%, and 19.3% respectively in the urban students; all significantly higher than those in the suburban students (4.5%, 4.3%, 12.5%, and 8.3% respectively, all P < 0.01). The rates of anamnestic nasal symptoms without common cold, present nasal symptoms without common cold, and those nasal symptoms complicated with itching eyes and lacrimation were 41.6%, 35.6%, and 10.3% respectively in urban students, and 21.3% all significantly higher than those in the suburban students (21.3%, 12.3%, and 3.3% respectively, all P < 0.05). The positive rates of the 13 main allergens were significantly higher in the urban students than in the suburban students (all P < 0.01). The positive rate of Dermatophagoides farinae and D. pteronyssinus, cat hair, and Blatella germanica were 20.4%, 16.9%, 10.8%, and 11.0% respectively in the urban students, all significantly higher than those in the suburban students (3.7%, 2.7%, 3.7%, and 2.7% respectively, all P < 0.01). CONCLUSION: The prevalence rates of allergic disorders have been increased in comparison with those in the last decade in Beijing junior middle school students, especially those in the urban area and may be due to the changes of environments and living styles.

Adolescent↗

PubMed2003

Therapeutic effects of ghrelin on endotoxic shock in rats.

We investigated the effects of ghrelin in a rat endotoxic shock model, and also observed the direct role of endotoxin on ghrelin generation in gastric mucosa. About 55% (11/20) of rats treated with lipopolysaccharide (5 mg/kg i.v.) alone died within 24 h of endotoxin injection. However, administration of ghrelin either at the same time as lipopolysaccharide injection (early treatment) or 12 h after lipopolysaccharide injection (late treatment) significantly decreased the mortality rate and ameliorated the hypotension seen in rats with endotoxic shock. Early and late treatment with ghrelin increased markedly the plasma glucose concentration and decreased the plasma lactate concentration. Early treatment with ghrelin attenuated significantly the deficiency in myocardial ATP content, but late treatment with ghrelin had no effect on myocardial ATP content. The plasma ghrelin level was significantly increased in the rats with endotoxin shock, and it increased further after ghrelin administration. Exposure of rat gastric mucosa in vitro to lipopolysaccharide (1.0 to 100 microg/ml) triggered the release of ghrelin from mucosa tissue in a dose- and time-dependent manner, meaning that lipopolysaccharide stimulated directly gastric mucosa to synthesize and secrete ghrelin. The results suggest that ghrelin could have therapeutic value for endotoxic shock.

Adenosine Triphosphate↗

PubMed2003

Controlled release of the herbicide, fluometuron, from matrix granules based on fractionated organosolv lignins.

An organosolv lignin and several of its fractions were used to prepare controlled release matrix granules with fluometuron using a melt process. Release profiles of fluometuron from these granules and a commercial granular formulation were studied in vitro. The release rates of fluometuron were considerably reduced from all lignin-based granules as compared to the commercial formulation. The release rates markedly varied with the lignin fraction used in the fluometuron-lignin matrix systems with the corresponding times for 50% release (T(50)) values ranging from 2.37 to 11.2 days. The variation in release rate of fluometuron was related to the high proportion of low molecular weight lignin and its associated water soluble lignin in the lignin-based granules, in terms of the hydrophobicity of matrix, the content, and release rate of soluble lignin. The release kinetics were fitted to the generalized model for up to 60% release of fluometuron; the kinetics of fluometuron were of the anomalous type for all matrixes in which release rates increased following a delay.

Chemical Fractionation↗

PubMed2003

Specific tumoricidal activity of a secreted proapoptotic protein consisting of HER2 antibody and constitutively active caspase-3.

In this study, a novel approach to antitumor therapy was devised by generating a chimeric tumor-targeted killer protein, referred to as immunocasp-3, that comprises a single-chain anti-erbB2/HER2 antibody with a NH(2)-terminal signal sequence, a Pseudomonas exotoxin A translocation domain, and a constitutively active caspase-3 molecule. In principle, cells transfected with the immunocasp-3 gene would express and secrete the chimeric protein, which then binds to HER2-overexpressing tumor cells. Subsequent cleavage of the constitutively active capase-3 domain from the immunocasp-3 molecule and its release from internalized vesicles would lead to apoptotic tumor cell death. To test this strategy, we transduced human lymphoma Jurkat cells with a chimeric immunocasp-3 gene expression vector and showed that they not only expressed and secreted the fusion protein but also selectively killed tumor cells overexpressing HER2 in vitro. i.v. injection of the transduced Jurkat cells led to tumor regression in a mouse xenograft model because of continuous secretion of immunocasp-3 by the transduced cells. The growth of HER2-positive tumor cells in this model was inhibited by i.m. as well as intratumor injection of immunocasp-3 expression plasmid DNA, indicating that the immunocasp-3 molecules secreted by transfected cells have systematic antitumor activity. We conclude that the immunocasp-3 molecule, combining the properties of a tumor-specific antibody with the proapoptotic activity of a caspase, has potent and selective antitumor activity, either as cell-based therapy or as a DNA vaccine. These findings provide a compelling rationale for therapeutic protocols designed for erbB2/HER2-positive tumors.

Amino Acid Sequence↗

PubMed2003

Comparative analysis of expression and secretion of placental leptin in mammals.

Increased plasma level of leptin appears to be a ubiquitous feature of pregnant mammals. The mechanisms by which leptin levels are increased may be species specific, however, with some species upregulating adipose leptin production and others expressing leptin in the placenta. Placental leptin expression was examined in representative species of the two most abundant mammalian orders, Rodentia and Chiroptera, and in cultured human choriocarcinoma (BeWo) cells. Leptin mRNA was expressed in BeWo cells and in placentas of Myotis lucifugus (little brown bat), Eptesicus fuscus (big brown bat), and Rattus norvegicus (laboratory rat), but not the common laboratory mouse Mus musculus. cAMP stimulated secretion of leptin from BeWo cells and also stimulated leptin mRNA expression in the cells. In addition to adipose and placental tissue, leptin transcript in M. lucifugus was detectable in heart, spleen, and liver, but not in lung, brain, and kidney. Hepatic expression was also observed in E. fuscus, but not in mice or rats, and did not appear to result from hepatic fat deposition. Leptin cDNA was cloned and sequenced from M. lucifugus placenta and shared up to 95% homology with other mammalian leptin cDNAs. It is concluded that 1) placental leptin expression and secretion are species-specific traits, 2) placental leptin production represents one of three major mechanisms for achieving high circulating maternal leptin levels during pregnancy, the others being upregulation of adipose leptin production and production of circulating leptin-binding proteins, and 3) hepatic leptin expression in pregnant insectivorous bats may be an adaptation resulting from the presence of extremely low amounts of subcutaneous fat during pregnancy and lactation in these species.

Adipose Tissue↗

PubMed2003

[Targeted tumor suppression by a secreted fusion protein consisting of anti- erbB2 antibody and reversed caspase-3 to SKBr3 cells].

OBJECTIVE: To investigate the targeted killing effect to SKBr3 cells due to the expression of a secreted fusion protein consisting of anti-erbB2 antibody and reversed caspase-3. METHODS: A recombinant plasmid pCMV-e23scFv-PEII-revcasp 3 was constructed by subcloning reversed caspase-3 gene to the downstream of anti-erbB2 antibody and transfected into Jurkat cells. The cell lines which secreted expressing fusion protein stably were selected. The fusion protein in media was detected by ELISA and the media was used to culture human breast cancer SKBr3 cells. The recombinant plasmids with liposomes was administrated to BALB/C nude mouses bearing SKBr3 tumor by intramuscular injection. The targetting effect of the recombinant fusion protein caspase-3 was detected by indirect immunofluorescence staining. RESULTS: Fusion protein can be expressed and secreted by Jurkat cells stably and kill SKBr3 cells. Significant prolonged survival time (prolonged by 72%) and inhibition of tumor growth in vivo (within inhibition ratio of 77%) were seen in the group administered with recombinant plasmids. Indirect immunofluorescence staining showed that the recombinant fusion protein caspase-3 has targetting effect. CONCLUSION: Secreted expression of the fusion protein consisting of anti-erbB2 antibody and reversed caspase-3 can targetedly induce SKBr3 cells to death.

Animals↗

PubMed2003

Cold-active serine alkaline protease from the psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization.

An extracellular protease was purified from a deep-sea psychrophilic bacterium strain DY-A which was identified as a Pseudomonas species. The optimal growth and protease-producing temperatures of the strain were all 10 degrees C, and the protease was secreted only at temperatures under 20 degrees C. The enzyme was most active at 40 degrees C and at pH 10.0. It was inhibited by phenylmethyl sulfonylfluoride and diisopropyl fluorophosphate, indicating that it is a serine protease. Chelators such as EDTA, EGTA, 1,10-phenanthroline and 2,2'-bipyridyl produced a decrease of activity. The enzyme was sensitive to denaturing agents such as SDS, urea, and guanidine HCl and resistant to thiol-containing reducing agents such as dithiotreitol. The enzyme was active towards N-succinyl-Ala-Ala-Pro-Phe- p-nitroanilide and N-succinyl-Ala-Ala-Pro-Leu- p-nitroanilide. The native molecular mass of the enzyme determined by native PAGE and SDS-PAGE was 25 kDa.

Chromatography, DEAE-Cellulose↗

PubMed2003

The interaction between ADAM 22 and 14-3-3zeta: regulation of cell adhesion and spreading.

The ADAM family consists of a number of transmembrane proteins that contain disintegrin-like and metalloproteinase-like domains. Therefore, ADAMs potentially have cell adhesion and protease activities. 14-3-3 proteins are a highly conserved family of cytoplasmic proteins that associate with several intracellular signaling molecules in the regulation of various cellular functions. Here we report the identification of a novel interaction between the ADAM 22 cytoplasmic tail and the 14-3-3zeta isoform by a yeast two-hybrid screen. The interaction between the ADAM 22 cytoplasmic tail and 14-3-3zeta was confirmed by an in vitro protein pull-down assay as well as by co-immunoprecipitation, and the binding sites were mapped to the 28 amino acid residues of the C-terminus of the ADAM 22 cytoplasmic tail. Furthermore, we found that overexpression of the ADAM 22 cytoplasmic tail in human SGH44 cells inhibited cell adhesion and spreading and that deletion or mutation of the binding site for 14-3-3zeta within the ADAM 22 cytoplasmic tail abolished the ability of the overexpressed cytoplasmic tail to alter cell adhesion and spreading. Taken together, these results for the first time demonstrate an association between ADAM 22 and a 14-3-3 protein and suggest a potential role for the 14-3-3zeta/ADAM 22 association in the regulation of cell adhesion and related signaling events.

14-3-3 Proteins↗

PubMed2003

Chromatin reorganization accompanying cellular dedifferentiation is associated with modifications of histone H3, redistribution of HP1, and activation of E2F-target genes.

The remarkable regeneration capacity of plant cells is based on their capability to dedifferentiate. We recently reported that cellular dedifferentiation proceeds through two distinct phases, each accompanied by chromatin decondensation: acquisition of competence for fate switch followed by a signal-dependent reentry into S phase. The purpose of this study was to (1) characterize changes in chromatin factors associated with chromatin decondensation, and (2) study the relationship between chromatin decondensation and transcriptional activation of pRb/E2F-regulated genes. We show that plant cells competent for fate switch display a disruption of nucleolar domain appearance associated with condensation of 18S ribosomal DNA, as well as modifications of histone H3 and redistribution of heterochromatin protein 1 (HP1). We further show that the pRb/E2F-target genes RNR2 and PCNA are condensed and silent in differentiated leaf cells but become decondensed, although not yet activated, as cells acquire competence for fate switch; transcriptional activation becomes evident during progression into S phase, concomitantly with pRb phosphorylation. We propose that chromatin reorganization is central for reversion of the differentiation process leading to resetting of the gene expression program and activation of silent genes.

Cell Cycle Proteins↗

PubMed2003

Differences between natriuretic peptide receptors in the olfactory bulb and hypothalamus from spontaneously hypertensive and normotensive rat brain.

Natriuretic peptide receptor-A (NPR-A) functional characteristics in the hypothalamus and olfactory bulb (OB) have been investigated in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). Autoradiographic studies demonstrate a decreased number of atrial natriuretic peptide (ANP) binding sites in the olfactory bulb and hypothalamus in SHR compared to WKY rats. We found that NPR-A showed a lower maximal binding capacity (B(max)) and higher affinity in SHR than in WKY rats both in the olfactory bulb and hypothalamus. However, despite the lower B(max) in SHR, both ANP(1-28) and ANP(5-25) stimulated similar or greater cGMP production than in WKY rats. These differences were found even before the development of hypertension. NPR-A in the olfactory bulb and hypothalamus from 3-week-old SHR showed a lower B(max) and K(d) and a higher cGMP production rate than in WKY rats, suggesting that these characteristics are intrinsic of NPR-A in SHR, instead of being a result of hypertension itself. The present study provides evidences for altered NPR-A receptor properties and function in the olfactory bulb and hypothalamus from SHR, which might be involved in the pathogenesis of hypertension.

Animals↗

PubMed2003

Hydrolysis of fibrinogen and plasminogen by immobilized earthworm fibrinolytic enzyme II from Eisenia fetida.

Earthworm fibrinolytic enzyme II (EFE-II) from Eisenia fetida has a broad hydrolytic specificity for peptide bonds. Our experiments show that EFE-II can hydrolyze the specific chromogenic substrates of thrombin (Chromozym TH), trypsin (Chromozym TRY) and elastase (Chromozym ELA). The Michaelis-Menten constant (K(m)) for Chromozym ELA (approximately 245 microM) is much higher than those for the thrombin (approximately 90 microM) and trypsin (approximately 60 microM) substrates. On the other hand, EFE-II is inhibited most strongly by soybean trypsin inhibitor (SBTI), and weakly inhibited by elastinal, suggesting that EFE-II has a trypsin-like activity. Degradation of plasminogen (PLg) and fibrinogen by EFE-II was investigated after EFE-II had been immobilized onto 1,1'-carboryl-diimidazole (CDI)-activated Sepharose CL-6B. The immobilized EFE-II has 55-60% activity of the native enzyme with a higher thermal and pH resistance. EFE-II cleaves PLg at four hydrolytic sites: Lys(77)-Arg(78), Arg(342)-Met(343), Ala(444)-Ala(445) and Arg(557)-Ile(558). The site Arg(557)-Ile(558) is also recognized and cleaved by tissue plasminogen activator (t-PA) and urokinase (UK), producing active plasmin. Cleaving Ala(444)-Ala(445) released mini-plasmin with secondary activity to hydrolyze fibrin. Immobilized EFE-II degrades not only the Aalpha chain of fibrinogen in the C-terminal region (like human neutrophil elastase, HNE), but also in the N-terminal region at the Val(21)-Glu(22) site.

Amino Acid Sequence↗

PubMed2003

Four new triterpene saponins from the seeds of Aesculus chinensis.

Two pairs of new geometrically isomeric triterpenoid saponins were isolated from the seeds of Aesculus chinensis and characterized as 28-acetyl-21-tigloylprotoaescigenin 3-O-[beta-D-xylopyranosyl (1 --> 2)] [beta-D-glucopyranosyl (1 --> 4)] [beta-D-glucopyranosiduronic acid (isoescin IIa, 1) and 28-acetyl-21-angeloylprotoaescigenin 3-O-[-beta-D-xylopyranosyl (1 --> 2)] [beta-D-glucopyranosyl (1 --> 4)] beta-D-glucopyranosiduronic acid (isoescin IIb, 2); 28-acetyl-21-tigloylbarringtogenol C 3-O-[beta-D-galactopyranosyl (1 --> 2)] [beta-D-glucopyranosyl (1 --> 4)] beta-D-glucopyranosiduronic acid (isoescin IIIa, 3) and 28-acetyl-21-angeloylbarringtogenol C 3-O-[beta-D-galactopyranosyl (1 --> 2)] [beta-D-glucopyranosyl (1 --> 4)] beta-D-glucopyranosiduronic acid (isoescin IIIb, 4). Their structures were established on the basis of spectroscopic and chemical evidence.

Aesculus↗