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Biomedical subjects

Wei Liao

Publications and source records attributed to Wei Liao.

At least 19 recordsLinked to original sources
PubMed2026

SDC1+ CAFs secreting CTGF drive tumour metastasis via FGFR3 signalling in cancers.

BACKGROUND: Cancer-associated fibroblasts (CAFs) are key stromal components of the tumour microenvironment (TME) that profoundly influence tumour progression. However, CAFs exhibit pronounced phenotypic and functional heterogeneity, and whether conserved CAF subtypes with shared functional hallmarks exist across different cancer types remains unclear. OBJECTIVE: We sought to uncover universal CAF subtypes that transcend tumour origins, defining their core molecular signatures and pro-tumorigenic functions within the TME. DESIGN: We constructed a pan-cancer CAF atlas through single-cell transcriptomic analysis of 554 specimens across 14 cancer types. To validate the findings, we performed further functional analyses, including in vitro migration and invasion assays, in vivo lymphatic metastasis models and mechanistic studies focusing on candidate signalling pathways. RESULTS: We identified a conserved syndecan 1 (SDC1) + CAF subset associated with advanced tumour stage and poor outcomes. These CAFs enhanced tumour cell migration and invasion in vitro and promoted lymphatic metastasis in vivo. This effect is mediated through connective tissue growth factor (CTGF) secretion, which activates fibroblast growth factor receptor 3 (FGFR3) signalling in tumour cells to induce epithelial-mesenchymal transition (EMT). Blocking CTGF or FGFR3 signalling abrogated these effects. We also found that kruppel like factor 6 (KLF6) directly regulates CTGF in SDC1+ CAFs, establishing a complete KLF6-CTGF-FGFR3 metastatic axis. CONCLUSIONS: Our study establishes SDC1+ CAFs as a universal, metastasis-promoting CAF subset across multiple cancer types and uncovers a novel KLF6-CTGF-FGFR3 axis that drives EMT and tumour dissemination. These findings provide mechanistic insight into CAF-tumour cell crosstalk and highlight actionable stromal targets for anti-metastatic therapies across diverse malignancies.

Humans↗
PubMed2006

A new approach of pellet formation of a filamentous fungus -Rhizopus oryzae.

The effects of inoculum and medium composition (i.e. potato dextrose broth as carbon source, soybean peptone, calcium carbonate, and metal ions) on pellet formation of Rhizopus oryzae ATCC 20344 have been studied. Metal ions were found to have a significant negative effect on pellet formation while soybean peptone had a positive effect. In addition, potato dextrose broth and calcium carbonate were beneficial to R. oryzae for growing small, smooth pellets during the culture. The study also demonstrated that an inoculum size of less than 1.5x10(9)spores/L had no significant influence on pellet formation although it had large impacts on pellet growth. Thus, a new approach to form pellets has been developed using only potato dextrose broth, soybean peptone, and calcium carbonate allowing for pellet size to be controlled by adjusting inoculum size and the concentrations of potato dextrose broth, soybean peptone, and calcium carbonate in the medium.

Biomass↗
PubMed2006

Proteasome inhibition induces differential heat shock protein response but not unfolded protein response in HepG2 cells.

Liver, a central organ responsible for the metabolism of carbohydrates, proteins, and lipoproteins, is exposed to various kinds of physiological, pathological, and environmental stresses. We hypothesized that blockage of proteasome degradation pathway induces heat shock protein (HSP) response and unfolded protein response in the liver cells. In this study, we have characterized cellular responses to proteasome inhibition in HepG2 cells, a well-differentiated human hepatoma cells. We found that proteasome inhibition induced differential response among cytosolic HSPs, that is, increased expression of HSP70, but no change in HSP40, HSC70, and HSP90. However, proteasome inhibition did not induce typical unfolded protein response as indicated by absence of stimulation of GRP78 and GRP94 proteins. Upon proteasome inhibition, inclusion bodies were accumulated, and ubiquitin-conjugated proteins appeared in insoluble fraction, together with HSP40, HSP70, HSC70, and HSP90. After proteasome inhibition, misfolded proteins were increased in the cytosol and in the ER compartment as evaluated by examining ubiquitin-conjugated proteins. However, essentially all ER-associated ubiquitin-conjugated proteins were located on the surface of the ER, which explains why proteasome inhibition does not induce unfolded protein response. In conclusion, proteasome inhibition induces differential HSP response, but not unfolded protein response in HepG2 cells. Our study also suggests that HSPs play important roles in directing proteasomal degradation and protein aggregate formation.

3T3-L1 Cells↗
PubMed2006

Studying the effects of reaction conditions on components of dairy manure and cellulose accumulation using dilute acid treatment.

The objectives of this study were to statistically study the effects of reaction conditions of temperature, acid concentration, and reaction time on manure components of cellulose, hemicellulose, lignin, and nitrogen during dilute acid treatment of dairy manure; and to further optimize the accumulation of cellulose for later enzymatic conversion to glucose. A 2(3) full factorial design was adopted to investigate the effects of the reaction conditions on each individual component and later followed by a 3-factor central composite design which was used to obtain the optimal conditions for cellulose accumulation. The results indicated that acid was the most important factor for changes of all the components. The results also showed that two other individual factors, reaction time and temperature, as well as the interactions among all three factors had significant influences on the changes. In addition, the optimal conditions for cellulose accumulation were 2.8h reaction time, 140 degrees C reaction temperature, and 1.0% acid concentration. Under these conditions cellulose content reached 31.0% while hemicellulose, lignin and nitrogen content were 3.2%, 20.8% and 2.4%, respectively.

Animals↗
PubMed2006

Knockdown of apolipoprotein B, an atherogenic apolipoprotein, in HepG2 cells by lentivirus-mediated siRNA.

ApoB is an important determinant of atherosclerosis susceptibility and a potential pharmaceutical target for lowering atherogenic lipoproteins. In the present study, we used a lentiviral vector to express short hairpin RNAs for inhibition of apoB production in HepG2 cells. We first demonstrated that lentivirus could efficiently deliver transgene into HepG2 cells by using GFP lentivirus. We then made three lentiviral siApoB constructs, two of which were highly efficient for silencing apoB expression in HepG2 cells. We showed that siApoB lentivirus specifically knocked down apoB but had no effects on other proteins such as apoAI and albumin. Consequently, the secretion of apoB was reduced markedly. The silencing effect of siApoB lentivirus appeared to be permanent. Knocking down apoB did not alter the expression of cytoplasmic stress proteins (HSP70 and HSP90) and their ER homologues (GRP78 and GRP94). Furthermore, neither IKKalpha and JNK nor phosphorylated IKK and JNK were increased in long-term apoB-deficient hepatocytes as compared to the control cells. Consistent with these findings, apoB-deficient hepatocytes responded to insulin to a similar extent as the control cells as determined by measuring insulin-induced phosphorylation of IRS and ERK. Our studies indicate that lentiviral siRNAs provide an excellent approach for delivering siRNA into HepG2 cells and may be used for gene therapy for hyperlipidemia.

Apolipoproteins B↗
PubMed2006

Lentiviral short hairpin ribonucleic acid-mediated knockdown of GLUT4 in 3T3-L1 adipocytes.

Adipose tissue is an important insulin target organ, and 3T3-L1 cells are a model cell line for adipocytes. In this study, we have used lentivirus-mediated short hairpin RNA (shRNA) for functional gene knockdown in 3T3-L1 adipocytes to assess the molecular mechanisms of insulin signaling. We chose to target GLUT4 to validate this approach. We showed that lentiviruses efficiently delivered transgenes and small interfering RNA (siRNA) into fully differentiated 3T3-L1 adipocytes. We established a strategy for identifying efficient siRNA sequences for gene knockdown by transfecting 293 cells with the target gene fluorescent fusion protein plasmid along with a plasmid that expresses shRNA. Using these methods, we identified highly efficient siGLUT4 sequences. We demonstrated that lentivirus-mediated shRNA against GLUT4 reduced endogenous GLUT4 expression to almost undetectable levels in 3T3-L1 adipocytes. Interestingly, insulin-stimulated glucose uptake was only reduced by 50-60%, suggesting that another glucose transporter mediates part of this effect. When siGLUT1 was introduced into GLUT4-deficient adipocytes, insulin-stimulated glucose uptake was essentially abolished, indicating that both GLUT4 and GLUT1 contribute to insulin-stimulated glucose transport in 3T3-L1 adipocytes. We also found that GLUT4 knockdown led to impaired insulin-responsive aminopeptidase protein expression that was dependent on whether GLUT4 was knocked down in the differentiating or differentiated stage. We further found that GLUT4 expression was not required for adipogenic differentiation but was necessary for full lipogenic capacity of differentiated adipocytes. These studies indicate that lentiviral shRNA constructs provide an excellent approach to deliver functional siRNAs into 3T3-L1 adipocytes for studying insulin signaling and adipocyte biology.

3T3-L1 Cells↗
PubMed2006

Novel probes for protein chip applications.

Protein microarrays (protein chips), due to their high throughput, have great potential to reduce the cycle time of drug discovery and to improve the efficiency of medical diagnostics. A great deal of effort has been made to facilitate the development of protein chips, such as the antibody microarrays. High-throughput fabrication of purified high-affinity probes is now one of the bottlenecks for protein microarrays. We discuss here three novel strategies to fabricate probes, namely, single chain antibodies displayed by phage, protein-oligonucleotide conjugates, and aptamers, which are promising in the protein chip technology.

Antibodies↗
PubMed2006

Optimization of L-(+)-lactic acid production using pelletized filamentous Rhizopus oryzae NRRL 395.

Lactic acid is used as a food additive for flavor and preservation and a precursor in the development of poly-lactic acid, a product used to make biodegradable plastics and textiles. Rhizopus oryzae NRRL 395 is known to be a strain that produces optically pure L-(+)-lactic acid. The morphology of Rhizopus cultures is complex, forming filamentous, clumps, and pellet mycelia. Different morphology growth has significant effects on lactic acid production. In bioreactors, the filamentous or clump mycelia increase the viscosity of the medium, wrap around impellers, and block the nutrient transportation, leading to a decrease in production efficiency and bioreactor performance. Growing fungi in pellet form can significantly improve these problems. In this study, factors that affect lactic acid production in pelletized flask cultures using R. oryzae NRRL 395 were investigated in detail. Completely randomized designs were used to determine the influence of culture temperature, time, concentration of glucose, and inoculum size. Lactic acid fermentation using clump and pellet morphologies were performed in a 5 L fermentor at the optimal values obtained from flask culture. Finally, fed-batch culture was used to enhance the lactate concentration in broth. The final lactate concentration of fed-batch culture reached 92 g/L. The data presented in the article can provide useful information on optimizing lactic acid production using alternative source materials.

Biofilms↗
PubMed2006

[Controlled study on treatment of cervical spondylopathy of the nerve root type with acupuncture, moxibustion and massage as main].

OBJECTIVE: To probe into clinical value of comprehensive program of acupuncture, moxibustion and massage as main for treatment of cervical spondylopathy of the nerve root type. METHODS: Five centers, single blind, randomized controlled method were used, 660 cases were divided into a treatment group of 317 cases and a control group of 311 cases. They were treated respectively with comprehensive program of acupuncture, moxibustion and massage as main, and comprehensive program of physical therapy as main. Establish syndrome detection scale and multiply dimensional effect assessment indexes, and evaluate the therapeutic effects and safety. RESULTS: The cured rate, the cured-markedly effective rate were 42.9%, 64.4% in the treatment group, respectively, better than 16.7%, 36.3% in the control group (P<0.01); after treatment of 2 weeks, clinical symptoms improved in the both groups, but the treatment group was better than the control group in the improvement degrees of neck-shoulder-limb pain, neck rigidity, abnormality of cervical anteflexion, etc. (P<0.01 or P<0.05); the treatment group was shorter than the control group in the time of producing the effect and therapeutic course (P<0.01). CONCLUSION: Comprehensive program of acupuncture, moxibustion and massage as main is safe and effective for treatment of cervical spondylopathy, with a better therapeutic effect compared with the comprehensive program of physical therapy.

Acupuncture Therapy↗
PubMed2005

Acid hydrolysis of fibers from dairy manure.

Concentrated acid hydrolysis of lignocellulosic materials is a conventional treatment process for the production of mono-sugars. However, this method has been proved ineffective and undesirable for the treatment of dairy manure due to the high nitrogen content of dairy manure and the environmental issues caused by the use of highly concentrated acid solution. In an effort to overcome these barriers, a modified acid hydrolysis process with short reaction time was introduced that involved a nitrogen-removing pretreatment followed by decrystallization with concentrated acid and then hydrolysis using dilute acid. The effects of nitrogen, acid concentration, reaction time, and temperature were investigated. A pretreated manure with a low nitrogen content of 1.3% was used as the substrate. The results indicated that the optimal conditions for fiber decrystallization were 75% acid concentration, 3:5 sample to acid ratio (weight basis), and 30 min of reaction time; while the optimal conditions for acid hydrolysis were 12.5% acid and 10% dry sample at 135 degrees C for 10 min. These conditions produced 26 g/L glucose at a yield of 84% and 11 g/L hemicellulose-sugars at a yield of 80%.

Acids↗
PubMed2005

Production of cellulase by Trichoderma reesei from dairy manure.

Cellulase production by the fungi Trichoderma reesei was studied using dairy manure as a substrate. Data showed that T. reesei RUT-C30 had higher cellulase production than T. reesei QM 9414 and that a homogenized manure, treated by a blender to reduce fiber size, led to higher cellulase production. The cellulase production was further optimized by growing T. reesei RUT-C30 on homogenized manure. The effects of manure concentration, pH, and temperature on cellulase production were investigated with optimal parameter values determined to be 10 g/l manure (dry basis), 25.5 degrees C, and pH 5.7, respectively. Elimination of CaCl2, MgSO4, nitrogen sources (NH4+ and urea) and trace elements (Fe2+, Zn2+, Co2+ and Mn2+) from the original salt solution had no negative influence on the cellulase production, while phosphate elimination did reduce cellulase production. Based on above results, the final medium composition was simplified with manure additives being KH2PO4, tween-80 and CoCl2 only. Using this medium composition and a reaction time of 6-8 days, a maximum cellulase production activity of 1.74 IU/ml of filter paper activity, 12.22 IU/ml of CMCase activity, and 0.0978 IU/ml of beta-glucosidase was obtained. This filter paper activity is the highest ever reported in cellulase production from agricultural wastes.

Animals↗
PubMed2005

Effects of hemicellulose and lignin on enzymatic hydrolysis of cellulose from dairy manure.

This study focused on the effect of hemicellulose and lignin on enzymatic hydrolysis of dairy manure and hydrolysis process optimization to improve sugar yield. It was found that hemicellulose and lignin in dairy manure, similar to their role in other lignocellulosic material, were major resistive factors to enzymatic hydrolysis and that the removal of either of them, or for best performance, both of them, improved the enzymatic hydrolysis of manure cellulose. This result combined with scanning electron microscope (SEM) pictures further proved that the accessibility of cellulose to cellulase was the most important feature to the hydrolysis. Quantitatively, fed-batch enzymatic hydrolysis of fiber without lignin and hemicellulose had a high glucose yield of 52% with respect to the glucose concentration of 17 g/L at a total enzyme loading of 1300 FPU/L and reaction time of 160 h, which was better than corresponding batch enzymatic hydrolysis.

Carbohydrates↗
PubMed2005

Production of cellulase/beta-glucosidase by the mixed fungi culture of Trichoderma reesei and Aspergillus phoenicis on dairy manure.

A cellulase production process was developed by growing the fungi Trichoderma reesei and Aspergillus phoenicis on dairy manure. T. reesei produced a high total cellulase titer (1.7 filter paper units [FPU]/mL, filter paper activity) in medium containing 10 g/L of manure (dry basis [w/w]), 2 g/L KH2PO4, 2 mL/L of Tween-80, and 2mg/L of CoCl2. However, beta-glucosidase activity in the T. reesei-enzyme system was very low. T. reesei was then cocultured with A. phoenicis to enhance the beta-glucosidase level. The mixed culture resulted in a relatively high level of total cellulase (1.54 FPU/mL) and beta-glucosidase (0.64 IU/mL). The ratio of beta-glucosidase activity to filter paper activity was 0.41, suitable for hydrolyzing manure cellulose. The crude enzyme broth from the mixed culture was used for hydrolyzing the manure cellulose, and the produced glucose was significantly (p < 0.01) higher than levels obtained by using the commercial enzyme or the enzyme broth of the pure culture T. reesei.

Animals↗
PubMed2004

Optimizing dilute acid hydrolysis of hemicellulose in a nitrogen-rich cellulosic material--dairy manure.

Effective dilute acid hydrolysis of dairy manure which contains roughly 12% hemicellulose on a dry matter basis can produce a variety of mono-sugars such as arabinose, xylose and galactose, as well as to further benefit utilization of cellulose in the manure. To enhance the effectiveness of this dilute acid hydrolysis, the effect of manure nitrogen content was studied because some reactions such as the browning reaction between amino acids and reducing sugars and acid-base reactions involving ammonia and acid interfere with the hydrolysis. Two dairy manure samples were used to study this nitrogen effect; the original manure and the pretreated manure derived from a solid/liquid separation pretreatment. The pretreated manure had a total nitrogen content of 1.3% dry matter (DM) while the original dairy manure had twice that amount with a total nitrogen content of 2.6% DM. Results found that the optimal conditions for hydrolysis of manure hemicellulose were 2 h reaction time, 1% sulfuric acid concentration, 135 degrees C, and 10% sample concentration using the pretreated dairy manure as raw material. Under these conditions the corresponding sugar yield from hemicellulose was 111% and sugar concentration in the solution reached 16.5 g/l. At the same time, the hydrolyzed solid had 43% DM of cellulose, which was much higher than both the original manure containing 22% and the pretreated manure with 32%.

Acids↗
PubMed2004

Hydrolysis of animal manure lignocellulosics for reducing sugar production.

Converting animal manure into value-added products provides a potential alternative for treatment and disposal of such materials. Lignocellulosics are a major component of animal manure and represent an undeveloped bioresource. In this work, a process was developed for hydrolyzing manure lignocellulosics into fermentable sugars. When raw dairy manure was pre-treated with 3% sulfuric acid at 110 degrees C for 1 h, hemicellulose was completely degraded into mainly arabinose, galactose and xylose. The pretreated materials were then treated with cellulolytic enzymes, Celluclast-1.5L and Novozyme-188, to hydrolyze the cellulose. The optimal enzyme loadings were identified as 13 FPU cellulase/g substrate and 5 IU beta-glucosidase/g substrate. The optimal temperature and pH were determined to be 46 degrees C and 4.8, respectively. A substrate concentration of 50 g/l favored both glucose concentration (in hydrolysate) and glucose yield (based on per 100 g manure). It was also found that a reduced particle size of 590-mum resulted in a high glucose yield with further decreases in particle size not increasing the yield. For each particle size investigated, the addition of 2% tween-80 resulted in at least 20% improvement in glucose yield. The optimized hydrolysis process achieved a glucose yield of 11.32 g/100 g manure, which corresponded to about 40% cellulose conversion.

Carbohydrates↗
PubMed2004

Simultaneous production of nisin and lactic acid from cheese whey: optimization of fermentation conditions through statistically based experimental designs.

A biorefinery process that utilizes cheese whey as substrate to simultaneously produce nisin, a natural food preservative, and lactic acid, a raw material for biopolymer production, was studied. The conditions for nisin biosynthesis and lactic acid coproduction by Lactococcus lactis subsp. lactis (ATCC 11454) in a whey-based medium were optimized using statistically based experimental designs. A Plackett-Burman design was applied to screen seven parameters for significant factors for the production of nisin and lactic acid. Nutrient supplements, including yeast extract, MgSO4, and KH2PO4, were found to be the significant factors affecting nisin and lactic acid formation. As a follow-up, a central-composite design was applied to optimize these factors. Second-order polynomial models were developed to quantify the relationship between nisin and lactic acid production and the variables. The optimal values of these variables were also determined. Finally, a verification experiment was performed to confirm the optimal values that were predicted by the models. The experimented results agreed well with the model prediction, giving a similar production of 19.3 g/L of lactic acid and 92.9 mg/L of nisin.

Cell Culture Techniques↗
PubMed2003

Hypermethylation of the Ink4b locus in murine myeloid leukemia and increased susceptibility to leukemia in p15(Ink4b)-deficient mice.

The Ink4b gene (Cdkn2b) encodes p15(Ink4b), a cyclin-dependent kinase inhibitor. It has been implicated in playing a role in the development of acute myeloid leukemia (AML) in man, since it is hypermethylated with high frequency. We provide evidence that the gene is a tumor suppressor for myeloid leukemia in mice. The evidence is twofold: (1) retrovirus-induced myeloid leukemias of the myelomonocytic phenotype were found to have hypermethylation of the 5' CpG island of the Ink4b gene, and this could be correlated with reduced mRNA expression, as demonstrated by TaqMan real-time PCR. p15(Ink4b) mRNA expression in a leukemia cell line, with hypermethylation at the locus, was induced following treatment with 5-aza-2'-deoxycytidine. (2) Targeted deletion of one allele in mice by removal of exon 2 increases their susceptibility to retrovirus-induced myeloid leukemia. Mice deficient in both alleles were not more susceptible to myeloid disease than those deficient in one allele, raising the possibility that there are opposing forces related to the development of myeloid leukemia in Ink4b null mice.

Animals↗