Biomedical subjects

Liqun He

Publications and source records attributed to Liqun He.

10 recordsLinked to original sources

PubMed2006

Development of a single mode electromagnetic resonant cavity for rewarming of cryopreserved biomaterials.

An electromagnetic (EM) heating system is developed to achieve the rapid and uniform warming of cryopreserved biomaterials. Using the heating system, a rectangular resonant cavity is excited in TE101 mode at frequencies near 434 MHz. In experiments, a spherical phantom of biomaterial with a diameter of 36 mm is placed at the center of the cavity. The phantom is first cooled down to about -80 degrees C within the cavity and then thawed by EM absorption. Results show that EM warming can produce much higher warming rate than conventional water-bath warming method. The spatial temperature distribution in the phantom during EM warming is also more uniform than that during the water-bath warming.

Adhesives↗

PubMed2006

Microarray analysis of blood microvessels from PDGF-B and PDGF-Rbeta mutant mice identifies novel markers for brain pericytes.

Normal blood microvessels are lined by pericytes, which contribute to microvessel development and stability through mechanisms that are poorly understood. Pericyte deficiency has been implicated in the pathogenesis of microvascular abnormalities associated with diabetes and tumors. However, the unambiguous identification of pericytes is still a problem because of cellular heterogeneity and few available molecular markers. Here we describe an approach to identify pericyte markers based on transcription profiling of pericyte-deficient brain microvessels isolated from platelet-derived growth factor (PDGF-B)-/- and PDGF beta receptor (PDGFRbeta)-/- mouse mutants. The approach was validated by the identification of known pericyte markers among the most down-regulated genes in PDGF-B-/- and PDGFRbeta-/- microvessels. Of candidates for novel pericyte markers, we selected ATP-sensitive potassium-channel Kir6.1 (also known as Kcnj8) and sulfonylurea receptor 2, (SUR2, also known as Abcc9), both part of the same channel complex, as well as delta homologue 1 (DLK1) for in situ hybridization, which demonstrated their specific expression in brain pericytes of mouse embryos. We also show that Kir6.1 is highly expressed in pericytes in brain but undetectable in pericytes in skin and heart. The three new brain pericyte markers are signaling molecules implicated in ion transport and intercellular signaling, potentially opening new windows on pericyte function in brain microvessels.

ATP-Binding Cassette Transporters↗

PubMed2006

Large-scale identification of genes implicated in kidney glomerulus development and function.

To advance our understanding of development, function and diseases in the kidney glomerulus, we have established and large-scale sequenced cDNA libraries from mouse glomeruli at different stages of development, resulting in a catalogue of 6053 different genes. The glomerular cDNA clones were arrayed and hybridized against a series of labeled targets from isolated glomeruli, non-glomerular kidney tissue, FACS-sorted podocytes and brain capillaries, which identified over 300 glomerular cell-enriched transcripts, some of which were further sublocalized to podocytes, mesangial cells and juxtaglomerular cells by in situ hybridization. For the earliest podocyte marker identified, Foxc2, knockout mice were used to analyze the role of this protein during glomerular development. We show that Foxc2 controls the expression of a distinct set of podocyte genes involved in podocyte differentiation and glomerular basement membrane maturation. The primary podocyte defects also cause abnormal differentiation and organization of the glomerular vascular cells. We surmise that studies on the other novel glomerulus-enriched transcripts identified in this study will provide new insight into glomerular development and pathomechanisms of disease.

Animals↗

PubMed2006

The structural and cellular viability in cryopreserved rabbit carotid arteries.

OBJECTIVE: We investigated the histological and mechanical changes in addition to viable cellular recovery in cryopreserved blood vessels. MATERIALS AND METHODS: Rabbit carotids were cryopreserved in a cryoprotective medium containing 1.5 M of 1,2-propanediol (PD) and then were thawed slowly in an ice bag that had been precooled in liquid nitrogen. Fresh carotids were used as the control. The fresh and freeze-thawed arteries were cultured for the growth of vascular smooth muscle cells (VSMCs). The freeze-thawed arterial tissues were perfused in vitro for 6, 12, or 24 h, respectively, to assess the integrity of carotid walls and the mechanical properties. RESULTS: The results showed that it took almost the same time (24 approximately 36 h) for the VSMCs of the PD-cryopreserved arteries to regenerate as those from the fresh arteries. Their growing speeds also were similar. On the contrary, Me2SO-cryopreserved (1.5 M) arteries were unable to regenerate VSMCs in culture. After freeze-thawing, the mechanical properties decreased significantly (P < 0.003 for elastic modulus and P < 0.001 for fracture strength). After in vitro perfusion of the freeze-thawed carotid arteries, all of the survived endothelial cells fell off, and some of the VSMCs denaturalized or necrosed. The internal elastic fibers and collagen showed various degrees of cracking. The mechanical properties were decreased (P < 0.05). CONCLUSION: Our findings demonstrate that the PD-containing cryoprotective medium can preserve regenerative capacity of VSMCs, which makes it a useful technique for viable VSMC recovery. However, the freeze-thawing process and the in vitro perfusion caused serious disruption in the arterial mechanical properties, rendering the cryopreserved blood vessels less useful for vessel reconstruction.

Animals↗

PubMed2004

Trapped water of human erythrocytes and its application in cryopreservation.

The novel differential scanning calorimetry method as a technique for determining human red cell volume during freezing process has been reexamined and has been shown to provide a final erythrocyte volume to be 53% of its isotonic value after freezing from 0 to -40 degrees C. A new type of electronic particle counter (Multisizer 3, Beckman Coulter Inc., USA) was used to measure cell volume changes in response to equilibration in anisotonic media, and which gave out an equilibrated volume to be 57% of cell isotonic value in solution of 3186 mOsm. Both of these results indicate that 34-40% of intracellular water is trapped and is unavailable for participation in osmotic shifts. These findings are consistent with the published data that at least 20-32% (v/v) of the isotonic cell water is retained within RBCs. Then the application of trapped water in both simulation of freezing models and freezing-drying control was pointed out.

Blood Preservation↗

PubMedDate not supplied

Neural cell adhesion molecule-deficient beta-cell tumorigenesis results in diminished extracellular matrix molecule expression and tumour cell-matrix adhesion.

To understand by which mechanism neural cell adhesion molecule (N-CAM) limits beta tumour cell disaggregation and dissemination, we searched for potential downstream genes of N-CAM during beta tumour cell progression by gene expression profiling. Here, we show that N-CAM-deficient beta-cell tumorigenesis is associated with changes in the expression of genes involved in cell-matrix adhesion and cytoskeletal dynamics, biological processes known to affect the invasive and metastatic behaviour of tumour cells. The extracellular matrix (ECM) molecules emerged as the primary target, i.e. N-CAM deficiency resulted in down-regulated mRNA expression of a broad range of ECM molecules. Consistent with this result, deficient deposition of major ECM stromal components, such as fibronectin, laminin 1 and collagen IV, was observed. Moreover, N-CAM-deficient tumour cells displayed defective matrix adhesion. These results offer a potential mechanism for tumour cell disaggregation during N-CAM-deficient beta tumour cell progression. Prospective consequences of these findings for the role of N-CAM in beta tumour cell dissemination are discussed.

Animals↗

PubMedDate not supplied

A modified differential scanning calorimetry for determination of cell volumetric change during the freezing process.

A modified analytical and experimental method using differential scanning calorimeter (DSC) was developed to determine the cell volume change during the freezing process. Two cell types were used in the study: human platelets and erythrocytes (red blood cells). Isotonic cell suspensions with different cytocrits were prepared and used in the DSC experiments. Low cooling rates were used to avoid intracellular ice formation. Cell suspensions were cooled from room temperature to -40 degrees C. Latent heat release from the freezing of cell suspensions was shown to be a linear function of cytocrit. From slope and intercept of the linear function, cell volume change was determined based on a developed theoretical model. From experimental data and theoretical analyses, it was revealed that (a) the final volume of a human platelet at -40 degrees C was 33.7% of its isotonic volume, and 15.2% of the original (at isotonic condition) intracellular water remained unfrozen inside platelets, and (b) the final volume of human erythrocyte at -40 degrees C was 50.0% of its isotonic volume, and 30.3% of the original intracellular water was kept inside cells as residual unfrozen water.

Blood Platelets↗

PubMedDate not supplied

A numerical study of cell behaviour in a ternary solution during the freezing process.

Using a continuum model for multi-component phase change system, the freezing of cell suspension in a ternary solution, H2O-NaCl-CPA (cryoprotective agent) inside a flat bag is investigated numerically in this study. The temperature and phase change history, intracellular water loss, and the volume change of the cells at different locations inside cell suspension are calculated. Numerical results reveal that although the sample boundary is cooled at a constant rate, different locations inside the sample experienced different temperature changes and cooling rates. The highest cooling rates occur at internal locations. The cell volume change is location-dependent.

Cell Physiological Phenomena↗

PubMedDate not supplied

The pertinence of expression of heat shock proteins (HSPs) to the efficacy of cryopreservation in HELAs.

HELAs (Hela cells, passed cells of human cervical carcinoma) were heat or cold treated (named heat or cold shock) and then resumed normal culture for 2, 4 or 8 hours respectively. The expressions of heat shock protein 70 (HSP70) and 90 (HSP90) of the HELAs were measured by Northern and Western blotting. HELAs after 4-hour culture were exposed to or cryopreserved with different concentration of dimethyl sulfoxide (Me2SO, 2.5%, 5%, 10%, 15% and 20% respectively, V/V). Meanwhile, the HELAs after different culture time (2, 4 and 6 hours of culture) were cryopreserved with 5% Me2SO. After exposure or cryopreservation, the number of live HELAs was counted and the survival rate was calculated. The results showed that heat shock increased the expression of HSP70 and HSP90 of HELAs, while cold shock decreased the expression of the two proteins. When the concentrations of Me2SO were 10%, 15% and 20%, the survival rates of HELAs after exposure to Me2SO or cryopreservation were much lower than those when the concentrations were small. The survival rates of the heat shocked HELAs were significantly higher than those of the cold shocked and control HELAs. After cryopreservation with 5% Me2SO, the survival rate of heat shocked HELAs group with 2 hours culture time was the lowest among all the groups of HELAs with different cultural time. From the results of this study, we conclude that the expressions of HSP70 and HSP90 in HELAs increased significantly after heat shock, while cold shock decreased the expressions of these two proteins. The over-expressions of HSPs in the heat shocked HELAs could protect the cells from both injury caused by potential toxicity of high concentrations of Me2SO and cryoinjury caused by the freeze-thawing/cryopreservation procedure.

Cell Survival↗

PubMedDate not supplied

Numerical simulation of tissue freezing by liquid nitrogen based cryoprobe.

A model is presented to simulate the cooling processes during tumor cryosurgery with different kinds of flows through the cryoprobe. The heat flux between the cryoprobe wall and the tumor, the heat transfer coefficient under different inflow conditions are obtained numerically. The impact of the inlet mass flow rate, gas volume fraction on these parameters is investigated. It is found that the heat transfer coefficient decreased significantly when inflow changed from two-phase annular flow to droplet flow, and to gas flow. The inlet gas volume fraction and flow velocity only significantly affect the freezing ability of the probe when the inflow is gas or in droplet phase. Simulation of the tumor temperature profiles under different flow conditions show that the heat transfer coefficient is a crucial parameter in temperature prediction during cryosurgery. Results indicate that when the cryoprobe wall is assumed at a constant temperature conventionally, the cooling effect could be overestimated. It would be more reasonable to use the constant wall heat transfer coefficient to simulate the cooling progress under a specific flow.

Cryosurgery↗