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Biomedical subjects

Y Z Zhang

Publications and source records attributed to Y Z Zhang.

At least 19 recordsLinked to original sources

Screening and mutagenesis of a novel Bacillus pumilus strain producing alkaline protease for dehairing.

AIMS: To characterize and optimize a novel Bacillus pumilus strain isolated from biological waste which produces protease with excellent dehairing effect. This newly isolated strain could be utilized in the industrial leather dehairing process. METHODS AND RESULTS: Bacterial strains secreting proteases were screened from biological wastes. Positive clones were further characterized by analysing their efficacy in dehairing and effects on collagen integrity. Among 171 colonies tested, a strain BA06, identified as B. pumilus, was picked owing to its efficient dehairing capabilities with minimal impact on collagen. By combined mutagenesis using UV, N-methyl-N'-nitro-N-nitrosdguanidine and Co(60)-gamma-rays, this strain was further improved with regard to its alkaline protease production. The alkaline protease activity of the mutant strain SCU11was greatly improved up to 6000 U ml(-1), in comparison with its parent strain BA06 of 1200 U ml(-1). CONCLUSIONS: By using screening and mutagenesis methods, we have successfully created a B. pumilus strain that can produce high levels of alkaline proteases that are able to efficiently remove hair from skin with minimal damage on the collagen. SIGNIFICANCE AND IMPACT OF THE STUDY: This strain could be used in commercial alkaline protease production for leather dehairing.

Animals↗

Development of an effective sample preparation approach for proteomic analysis of silkworm eggs using two-dimensional gel electrophoresis and mass spectrometry.

Sample preparation is still the first and important step toward successful two-dimensional gel electrophoresis (2DE) and identification in proteomics study. The 2DE profiling of eggs of silkworm species by using conventional one-step extraction, however, is unsatisfactory because high-abundance proteins such as egg-specific protein (ESP) and No 30 family (30 KP) in the extract lead to difficulties in detecting most of biologically relevant proteins. Based on the tendency of these abundant proteins to be soluble in Tris-HCl buffer, we report herein a robust approach in which the extract enriched in ESP and 30 KP was fractionationed and mixed with the re-extract of residual pellet in an optimal proportion. In comparison with the one-step method, the 2DE pattern was improved by this new method with over one-third enhancement in spots. A total of 48 unique proteins obtained have been furthermore identified by mass spectrometry (MS) and MS/MS. The identified proteins are found to include heat shock proteins families, ribosomal proteins, disulfide isomerase proteins, Glutathione S-transferase, and elongation factor, etc., which are mainly involved in some important processes. To our knowledge, this is the first time that the several proteins have been detected in silkworm eggs by proteomics means. This simple and reproducible approach would raise the opportunity of discovering and identifying more biomarkers and determining their possible roles in further studies.

Animals↗

Coaxial electrospinning of (fluorescein isothiocyanate-conjugated bovine serum albumin)-encapsulated poly(epsilon-caprolactone) nanofibers for sustained release.

As an aim toward developing biologically mimetic and functional nanofiber-based tissue engineering scaffolds, we demonstrated the encapsulation of a model protein, fluorescein isothiocyanate-conjugated bovine serum albumin (fitcBSA), along with a water-soluble polymer, poly(ethylene glycol) (PEG), within the biodegradable poly(epsilon-caprolactone) (PCL) nanofibers using a coaxial electrospinning technique. By variation of the inner flow rates from 0.2 to 0.6 mL/h with a constant outer flow rate of 1.8 mL/h, fitcBSA loadings of 0.85-2.17 mg/g of nanofibrous membranes were prepared. Variation of flow rates also resulted in increases of fiber sizes from ca. 270 nm to 380 nm. The encapsulation of fitcBSA/PEG within PCL was subsequently characterized by laser confocal scanning microscopy, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS) analysis. In vitro release studies were conducted to evaluate sustained release potential of the core-sheath-structured composite nanofiber PCL-r-fitcBSA/PEG. As a negative control, composite nanofiber PCL/fitcBSA/PEG blend was prepared from a normal electrospinning method. It was found that core-sheath nanofibers PCL-r-fitcBSA/PEG pronouncedly alleviated the initial burst release for higher protein loading and gave better sustainability compared to that of PCL/fitcBSA/PEG nanofibers. The present study would provide a basis for further design and optimization of processing conditions to control the nanostructure of core-sheath composite nanofibers and ultimately achieve desired release kinetics of bioactive proteins (e.g., growth factors) for practical tissue engineering applications.

Animals↗

Development and assessment of conventional and targeted drug combinations for use in the treatment of aggressive breast cancers.

Combination chemotherapy has been at the forefront of cancer treatment for over 40 years. However, the rationale for selecting drug combinations and the process used to demonstrate clinical effectiveness has primarily followed trial and error methodology. Typically, the selection and assessment of combined drug therapies has been based on the effectiveness of each agent as monotherapy in treating the neoplasm and avoiding overlapping toxicities, followed by clinical trials to establish dose scheduling, toxicity, and efficacy. Unfortunately, this scheme is inefficient in terms of the time required to complete and revise these clinical trials based on the outcome to optimize the drug combination. A more rational approach for the development of combination oncology products should consider (i) in vitro assays for assessing therapeutic effects of drug combinations (antagonistic, additive or synergistic interactions) when added simultaneously; (ii) methods for measuring these interactions in vivo; (iii) the importance of understanding pharmacokinetic and biodistribution parameters when using drug combinations; (iv) the need to assess pathways known to contribute to cancer cell survival as well as metastasis; and (iv) the need to assess the fate of different cell populations (cancer and stroma) contributing to the development of cancer. Therefore, the goal of this article is to provide a road map for the preclinical development of drug combination products that will have improved therapeutic activity and a high likelihood of providing beneficial therapeutic outcomes in patients with aggressive cancers with a specific focus on patients with breast cancer.

Animals↗

Toxicity and protective efficiency of cryoprotectants to flounder (Paralichthys olivaceus) embryos.

With the purpose of finding an ideal cryoprotectant or combination of cryoprotectants in a suitable concentration for flounder (Paralichthys olivaceus) embryo cryopreservation, we tested the toxicities, at culture temperature (16 degrees C), of five most commonly used cryoprotectants-dimethyl sulfoxide (Me2SO), glycerol, methanol (MeOH), 1,2-propylene glycol (PG) and ethylene glycol (EG). In addition, cryoprotective efficiency to flounder embryos of individual and combined cryoprotectants were tested at -15 degrees C for 60 min. Five different concentrations of each of the five cryoprotectants and 20 different combinations of these cryoprotectants were tested for their protective efficiency. The results showed that the toxicity to flounder embryos of the five cryoprotectants are in the following sequence: PG < MeOH < Me2SO < glycerol < EG (P < 0.05); whereas the protective efficiency of each cryoprotectant, at -15 degrees C for a period of 60 min, are in the following sequence: PG > Me2SO approximately MeOH approximately glycerol > EG (greater symbols mean P < 0.05, and approximate symbols mean P > 0.05). Methanol combined with any one of the other cryoprotectants gave the best protection, while ethylene glycol combined with any one of the other cryoprotectants gave the poorest protection at -15 degrees C. Toxicity effect was concentration dependent with the lowest concentration being the least toxic for all five cryoprotectants at 16 degrees C. For PG, MeOH and glycerol, 20% solutions gave the best protection at -15 degrees C; whereas a 15% solution of Me2SO, and a 10% solution of EG, gave the best protection at -15 degrees C.

Animals↗

Dimerization in a half-filled one-dimensional extended hubbard model.

We use a density-matrix renormalization group method to study quantitatively the phase diagram of a one-dimensional extended Hubbard model at half filling by investigating the correlation functions and structure factors. We confirmed the existence of a novel narrow region with long-range bond-order-wave order that was highly controversial recently between the charge-density-wave phase and Mott insulator phase. We determined accurately the position of the bicritical point U(b) approximately 7.2t, V(b) approximately 3.746t, which is quite different from previous studies.

Journal Article↗

Novel concentration-killing curve method for estimation of bactericidal potency of antibiotics in an in vitro dynamic model.

The bactericidal pharmacodynamics of antibiotics against Escherichia coli were analyzed by a concentration-killing curve (CKC) approach, and the novel parameters median bactericidal concentration (BC(50)) and bactericidal intensity (r) for bactericidal potency were proposed. By using the agar plate method, about 500 E. coli cells were inoculated onto Luria-Bertani plates containing a series of antibiotic concentrations, and after 24 h of incubation at 37 degrees C, all the viable colonies were enumerated. This resulted in a sigmoidal CKC that could be perfectly fitted (R(2) > 0.9) with the function N = N(0)/[1 + e(r(x - BC(50)))], where N is number of colonies surviving on each plate with an x series of concentrations of an antibiotic, and N(0) represents the meaningful inoculum size. Construction of the CKC method was based on the bactericidal effect of each antibiotic against the bacterial strain versus the concentration in two dimensions and may be a more valid, accurate, and reproducible method for estimating the bactericidal effect than the endpoint minimum bactericidal concentration (MBC) method. Mathematically, the CKC approach was point symmetrical toward its inflexion (BC(50), N(0)/2); thus, 2BC(50) could replace MBC. The parameter BC(1) can be defined as BC(50) + [ln(N(0) - 1)/r], which is the drug concentration at which only one colony survived and which is the least critical value of MBC in the CKC. The variate r, which determined the tangent slope on inflexion when N(0) was limited, could estimate the bactericidal intensity of an antibiotic. This verified that the CKC approach may be useful in studies with other classes of antibiotics and has considerable value as a tool for the accurate and proper administration of antibiotics.

Algorithms↗

Cryopreservation of flounder (Paralichthys olivaceus) sperm with a practical methodology.

A simple and convenient protocol for the cryopreservation of the flounder (Paralichthys olivaceus) sperm was established for "on the spot" cryopreservation of large quantities of semen. The use of three cryoprotectants, dimethyl sulphoxide (DMSO), glycerol (Gly) and methanol was tested in the method. The percentage of motile sperm present in semen after it had been frozen and thawed in the presence of DMSO, Gly or methanol was 60.5+/-3.6, 79.17+/-4.5 and 13.25+/-4.7%, respectively. The fertilization rates of this sperm were 67.06+/-15.1, 76.20+/-10.0 and 44.93+/-22.6%, while the hatching rates of eggs fertilized with this sperm were 37.40+/-8.3, 48.18+/-25.7 and 23.35+/-10.8%, respectively. It was found that Gly and DMSO were better cryoprotectants than methanol, with Gly giving the best overall results. Under scanning electron microscopy, it could be seen that while the majority of the frozen-thawed sperm remained morphologically normal, some exhibited lost or dilated mitochondria, swollen mid-pieces, broken tails, or damaged cell membrane, which probably caused the decrease in motility and fertility of the frozen-thawed sperm.

Animals↗

Synthesis and evaluation of novel pseudomycin side-chain analogues. Part 3.

To increase the therapeutic utility of C-18 side-chain bearing pseudomycin analogue 2, we prepared additional analogues and prodrugs of 2 containing further modifications at various positions within its core structure. Each of the newly synthesized derivatives (10-15) exhibited reduced tail vein toxicity relative to the parent compound. Some of the new pseudomycin derivatives (e.g., 14) also showed improved in vivo antifungal activity relative to its corresponding parent compound.

Animals↗

Genetic progression and heterogeneity associated with the development of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma is a common fatal cancer, and Shanxi province, a region in north-central China, has some of the highest esophageal cancer rates in the world. Chromosomal regions with frequent allelic loss may point to major susceptibility genes that will assist us in understanding the molecular events involved in esophageal carcinogenesis and may serve as the basis for the development of markers for genetic susceptibility and screening for early detection of this cancer. This study was designed to identify events in the molecular progression of precursor and invasive lesions of squamous esophageal cancer. Twelve marker loci identified during our previous studies as having some of the highest rates of loss of heterozygosity (LOH) in invasive esophageal cancer were evaluated in laser-microdissected DNA obtained from low- and high-grade dysplastic lesions and invasive tumor foci from 10 fully embedded esophageal resection specimens. Each resection specimen contained a spectrum of disease, from epithelium that appeared histologically normal to invasive cancer, including a single dominant tumor surrounded by a region of precursor lesions (low- and high-grade dysplasia) and occasional "remote," nonadjacent precancerous foci. Using the 12 polymorphic markers, LOH was found in all of the three stages of disease. The frequency of LOH for all of the markers together increased with increasing disease severity. Among the informative low-grade dysplasia samples, LOH was detected with markers D3S1766 (3p), D4S2632 (4p), D9S910 (9q), and D13S1493 (13q), suggesting that LOH at these loci may be associated with early stages of tumor initiation and/or progression. LOH was detected among the informative high-grade (but not low-grade) dysplasia samples for the other eight markers tested, suggesting that LOH at these loci may occur later in the neoplastic process. In addition to the association between disease progression and these genetic changes, considerable genetic heterogeneity was found in each fully embedded resection specimen both between and within geographically separate neoplastic lesions.

Adult↗

Syntheses and antifungal activities of novel 3-amido bearing pseudomycin analogues.

As a result of our core SAR effort, we discovered a large number of 3-amido pseudomycin B (PSB) analogues (e.g., 4e LY448212 and 5b LY448731) that retain good in vitro and in vivo (IP) activities against Candida and Cryptococcus without inherent tail vein irritation. Several dimethylamino termini bearing 3-amides (e.g., 5b) also exhibited improved potency against Aspergillus in vitro. When evaluated in a two-week rat toxicology study, it was found that all animals receiving 4e (up to 75 mg/kg) were found to be normal. On the basis of these observations, we are convinced that it is possible to broaden the antifungal spectrum and improve the safety profile of pseudomycin analogues at the same time.

Amides↗

8-Amido-Bearing pseudomycin B (PSB) analogue: novel antifungal agents.

During the course of a structure-activity relationship (SAR) study on novel depsinonapeptide pseudomycin B, we synthesized a total of 12 8-amidopseudomycin analogues via standard two-step sequence from either ZPSB 2 or AllocPSB 3. A number of these amides exhibited good in vitro antifungal activities.

Animals↗

[Effect of costal perichondrium on regeneration and remodeling of costal neocartilage].

OBJECTIVE: To provide experimental basis for improving the curative effect of pectus excavatum. METHODS: Twelve rabbits were adopted in this experiment. After the bilateral second and third costal cartilages of the rabbits were resected subperichondrially, their right second and third costal perichondriums were damaged intentionally. Then, the bilateral third costal perichondriums were stitched into a tube-like structure and the second ones were left opened. After 2, 4, 6 of operation, the bilateral second and third neocartilages were measured for their width, and histological character were observed under microscope. RESULTS: 1. After 2, 4, 6 months of operation, the average width of the bilateral second neocartilages were significantly greater than the preoperative ones. 2. 4 and 6 months after operations, there was no significant difference in the average width of the bilateral third neocartilages and the preoperative ones. 3. The amount, distribution of costal neocartilage cells and the arrangement of costal neocartilage matrix within the left second and third costal cartilages were better than the right under the light microscope. 4. The left third costal neocartilage was regenerated and remodeled better than all the others. CONCLUSION: The integrality of costal perichondrium is in favor of the regeneration of costal cartilage, and the sleeve stitch of costal perichondrium facilitates the remodeling of costal neocartilage.

Animals↗

[Studies on lignocellulolytic enzymes production and biomass degradation of Pleurotus sp2 and Trametes gallica in wheat straw cultures].

Pleurotus sp2 and Trametes gallica were selected in this assay because of their high activities of lignocellulolytic enzymes and the enzyme peaks appeared at the early stage of liquid state fermentation. Solid state fermentation was also investigated for their abilities and behaviors of enzyme-production. The capabilities and characteristics of the two strains in degrading biomass were studied. When Pleurotus sp2 was incubated in wheat straw powder containing the liquid medium of low-nitrogen, no-carbon and high inorganic salt, the activities of MnP and Lac reached the peaks on the tenth day, but the activities of hemicellulases reached the peak on the 40th day. Pleurotus sp2 caused 17.6% of biomass loss. When T. gallica was incubated in wheat straw powder containing the liquid medium of hlig-nitrogen, or low-nitrogen, no-carbon and high inorganic salt, the activities of MnP reached the peaks on the tenth day, the lac and hemicelluloses on the 40th day, and the lignin peroxidases reached the peaks on the 50th day, and it caused more than 64% of biomass loss. Among them the hemicellulose was degraded by 71.96%, and the cellulose 66.21%. T. gallica was very capable of degrading lignin of wheat straw and caused 34.37% loss during 20 days, 46. 71% loss during 30 days and 70.14% loss during 60 days. It was interesting that T. gallica degraded lignin preferentially with respect to cellulose, which was very beneficial to biopulping of paper industry.

Basidiomycota↗

[Study on effect of shenmai injection in protecting myocardium against ischemia-reperfusion injury in thrombolytic therapy with urokinase for acute myocardial infarction patients evaluated by 99mTc-MIBI myocardial imaging].

OBJECTIVE: To evaluate the myocardial protecting effect of Shenmai injection (SMI) against ischemia/reperfusion injury in thrombolytic therapy with urokinase (UK) for acute myocardial infarction patients by 99mTc-MIBI myocardial imaging (SPECT). METHODS: Five hundred and thirty-seven patients were randomly divided into two groups. The SMI group (n = 292) was treated with thrombolytic treatment plus SMI and the control group (n = 245) with thrombolytic treatment only. Single photon emission computerized tomography (SPECT) was carried out on the 7th day after thrombolysis to determine the ischemic myocardial area (IMA) and ejection fraction (EF) in both groups and compared. RESULTS: The reperfusion rate of infarction related area (IRA) in the two groups was not different significantly (72.26% vs 72.65%, P > 0.05). The IMA in patients of the SMI group, no matter with or without reperfused IRA (211 cases and 81 cases) respectively, was significantly lower than that in the control group (178 cases and 67 cases) respectively, P < 0.01 and P < 0.05 respectively. The EF value in the SMI group was significantly higher than that in the control group (P < 0.01). CONCLUSION: Using SMI in early stage of thrombolytic treatment in acute myocardial infarction could significantly reduce IMA and increase EF. SMI showed good protective effect against myocardial ischemia/reperfusion injury in thrombolytic treatment.

Adult↗

[Evaluation of cerebral circulatory compensation after carotid artery occlusion in rabbits].

OBJECTIVE: To observe the cerebral circular changes of rabbits during brain ischemia caused by carotid artery occlusion (CAO) with a loading test-cerebral vascular CO2 reactivity (CVR) technique. METHODS: Sixty white rabbits were divided into 6 groups, before and during 0-50 days after CAO, we studied the regional cerebral blood flow (rCBF) and CVR of frontal lobe on the same side, using laser doppler perfusion imager (LDI). RESULT: The rCBF-lesion caused by CAO could hardly be observed (P > 0.5), but CVR cut down obviously (P < 0.01). CONCLUSIONS: The loading test-cerebral vascular CO2 reactivity (CVR) allow the hemodynamic effect of a CAO on the intracerebral circulation to be observed before the reduced rCBF can be detected.

Animals↗

The inhibitory mechanisms of amlodipine in human vascular smooth muscle cell proliferation.

The abnormal proliferation of vascular smooth muscle cells (VSMCs) is closely related to vascular diseases. There is growing evidence that calcium antagonists inhibit VSMC growth/proliferation, yet their molecular mechanisms remain to be determined. Recent reports suggest that p42/p44 mitogen-activated protein kinases (MAPKs) play an important role in cell growth and proliferation induced by growth factors. This study was designed to determine whether these MAPKs are involved in VSMC proliferation induced by basic fibroblast growth factor (bFGF) and to examine the inhibitory effect of amlodipine. Human VSMCs were obtained from inner mammary artery. p42/p44 MAPKs activity was measured by immunoblotting assay using anti-p42/p44 phospho-MAPK antibody. 1) bFGF (20 ng/ml) significantly activated p42/p44 MAPKs with a peak time of 5-15 min, which was maintained for 3 h. PD98059 (100 nM-10 microM), a specific inhibitor of MAPK kinase, inhibited bFGF-induced p42/p44 MAPKs activation in a dose-dependent manner. 2) Amlodipine (1-100 nM) dose-dependently inhibited p42/p44 MAPKs activation by bFGF. 3) Amlodipine (10 nM) could inhibit both short-term and long-term p42/p44 MAPKs activation by bFGF. Our results indicate that bFGF could activate p42/p44 MAPKs. Amlodipine, which could inhibit bFGF-induced human VSMC proliferation, inhibited both short-term and sustained p42/p44 MAPKs activation by bFGF, suggesting that bFGF-induced VSMC proliferation may be related to p42/p44 MAPKs activation, and that the antiproliferative effect of amlodipine may be related to its inhibition of p42/p44 MAPKs activation.

Amlodipine↗

[Effect of dissolved oxygen on mutanolysin fermentation].

Effects of several parameters relating to dissolved oxygen(DO) on mutanolysin fermentation were studied. The experiment using shake flasks shows that the medium volume and shaker agitation speed affect the production of mutanolysin. At the same time, the agitation rate together with aeation rate has effects on DO in fermentor. Mutanolysin fermentation was affected by DO greatly. Oxygen is a key restricted factor in mutanolysin fermentation. It affects the metablism and physiological action of Streptomyces globisporus S186. Whatever the DO is excessive high or low, it won't benefit the mutanolysin production. If DO is super, S. globisporus S186 will grow luxuriantly but do not produce mutanolysin, while if DO is lower, the S. globisporus S186 won't grow well even not to produce mutanolysin. During the course of fermentation, the DO changed regularly. It is similar to many antibiotic fermentation and some amino acid fermentation. As S. globisporus S186 grow in exponential phase, DO begin to decrease rapidly from 6 h and get to the lowest point at 40 h or so. Subsequently mutanolysin starts to be produced. DO rises again from 90 h. The key technoloyg of oxygen control in the fermentation is to keep the DO at a suboptimum level. In order to get a high mutanolysin yield, during the culture in fermentor the agitation rate and aeration rate should be kept at 200 r/min and 1:0.8(V:V) respectively.

Endopeptidases↗