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Featured researches published by Guoliang Lv.


Biotechnology and Bioengineering | 2011

Bioartificial liver system based on choanoid fluidized bed bioreactor improve the survival time of fulminant hepatic failure pigs

Guoliang Lv; Lifu Zhao; Anye Zhang; Weibo Du; Yu Chen; Chengbo Yu; Xiaoping Pan; Yimin Zhang; Tao Song; Jiansheng Xu; Lanjuan Li

Bioartificial liver (BAL) support system has been proposed as potential treatment method for end‐stage liver diseases. We described an improved BAL system based on a choanoid fluidized bed bioreactor containing alginate–chitosan encapsulated primary porcine hepatocytes. The feasibility, safety, and efficiency of this device were estimated using an allogeneic fulminant hepatic failure (FHF) model. FHF was induced with intravenous administration of D‐galactosamine. Thirty FHF pigs were divided into three groups: (1) an FHF group which was only given intensive care; (2) a sham BAL group which was treated with the BAL system with empty encapsulation, and (3) a BAL group which was treated with the BAL system containing encapsulated freshly isolated primary porcine hepatocytes. The survival times and biochemical parameters of these animals were measured, and properties of the encapsulations and hepatocytes before and after perfusion were also evaluated. Compared to the two control groups, the BAL‐treated group had prolonged the survival time and decreased the blood lactate levels, blood glucose, and amino acids remained stable. No obvious ruptured beads or statistical decline in viability or function of encapsulated hepatocytes were observed. This new fluidized bed BAL system is safe and efficient. It may represent a feasible alternative in the treatment of liver failure. Biotechnol. Bioeng. 2011;108:2229–2236.


Transplantation Proceedings | 2010

Establishment and Characterization of Immortalized Porcine Hepatocytes for the Study of Hepatocyte Xenotransplantation

X. Pan; Weibo Du; Xiaopeng Yu; G. Sheng; Hongcui Cao; Chengbo Yu; Guoliang Lv; He Huang; Yunbo Chen; J. Li; Lanjuan Li

BACKGROUND In light of the critical shortage of donor livers, xenogeneic sources offer the best alternative to human hepatocytes for the treatment of acute liver failure. This study investigated whether a combination of simian virus 40 large T antigen (SV40 LT) and human telomerase catalytic subunit (hTERT) genes could immortalize primary porcine hepatocytes that could reverse acute liver failure (ALF) in rats. METHODS We cotransfected SV40 LT and hTERT genes into primary porcine hepatocytes to examine the features of the transfected cell lines. We characterized the potentially therapeutic effect of immortalized porcine hepatocytes in a rat model of ALF induced by 90% hepatectomy. RESULTS An immortalized porcine hepatocyte cell line, HepLi, was expanded by >250 passages. HepLi cells maintained the defining characteristics of primary porcine hepatocytes, including porcine albumin secretion, urea production, and diazepam metabolism. Intrasplenic transplantation of HepLi cells significantly improved liver function, and significantly prolonging the survival of rats with ALF. CONCLUSIONS Cotransfection of SV40 LT and hTERT immortalized primary porcine hepatocytes without tumorigenicity in vitro. The Immortalized porcine hepatocytes served as a potential cell resource for xenotransplantation.


International Journal of Artificial Organs | 2009

In vitro large-scale cultivation and evaluation of microencapsulated immortalized human hepatocytes (HepLL) in roller bottles.

Chengbo Yu; Guoliang Lv; Xiaoping Pan; Yanshan Chen; Hongcui Cao; Yiming Zhang; Weibo Du; Shigui Yang; Lanjuan Li

Background/Aims Microencapsulated hepatocytes have been proposed as promising bioactive agents for packed-bed or fluidized-bed bioartificial liver assist devices (BLADs) and for hepatocyte transplantation because of the potential advantages they offer of high mass transport rate and an optimal microenvironment for hepatocyte culture. We developed a large-scale and high-production alginate-chitosan (AC) microcapsule roller bottle culture system for the encapsulation of HepLL immortalized human hepatocytes. In this study, the efficacy of upscaling encapsulated HepLL cells production with roller bottle cultivation was evaluated in vitro. Methods Microencapsulated HepLL cells were grown at high yield in large-scale roller bottles, with free cells cultured in roller bottle spinners serving as controls. The mechanical stability and the permeability of the AC microcapsules were investigated, and the growth, metabolism and functions of the encapsulated HepLL cells were evaluated as compared to free cells. Results The microcapsules withstood well the shear stress induced by high agitation rates. The microcapsules were permeable to albumin, but prevented the release of immunoglobulins. Culture in roller bottles of immortalized human hepatocytes immobilized in the AC microcapsules improved cell growth, albumin synthesis, ammonia elimination and lidocaine clearance as compared with free cells cultured in roller bottles. Conclusions Encapsulated HepLL cells may be cultured on a large scale in roller bottles. This makes them possible candidates for use in cell-based liver assist therapies.


Vaccine | 2009

Construction and cellular immune response induction of HA-based alphavirus replicon vaccines against human-avian influenza (H5N1).

Shigui Yang; Jianer Wo; Minwei Li; Fen-fang Mi; Chengbo Yu; Guoliang Lv; Hongcui Cao; Haifeng Lu; Baohong Wang; Hanping Zhu; Lanjuan Li

Several approaches are being taken worldwide to develop vaccines against H5N1 viruses; most of them, however, pose both practical and immunological challenges. One potential strategy for improving the immunogenicity of vaccines involves the use of alphavirus replicons and VP22, a herpes simplex type 1 (HSV-1) protein. In this study, we analysed the antigenic peptides and homogeneity of the HA sequences (human isolates of the H5N1 subtype, from 1997 to 2003) and explored a novel alphavirus replicon system of VP22 fused with HA, to assess whether the immunogenicity of an HA-based replicon vaccine could be induced and augmented via fusion with VP22. Further, replicon particles expressing VP22, and enhanced green fluorescent protein (EGFP) were individually used as controls. Cellular immune responses in mice immunised with replicons were evaluated by identifying specific intracellular cytokine production with flow cytometry (FCM). Animal-based experimentation indicated that both the IL-4 expression of CD4(+) T cells and the IFN-gamma expression of CD8(+) T cells were significantly increased in mice immunised with VPR-HA and VPR-VP22/HA. A dose titration effect vis-à-vis both IL-4 expression and IFN-gamma expression were observed in VPR-HA- and VPR-VP22/HA-vaccinated mice. Our results revealed that both VPR-VP22/HA and VPR-HA replicon particles presented a promising approach for developing vaccines against human-avian influenza, and VP22 could enhance the immunogenicity of the HA antigens to which it is fused.


Scientific Reports | 2016

Efficacy of Fluidized Bed Bioartificial Liver in Treating Fulminant Hepatic Failure in Pigs: A Metabolomics Study.

Pengcheng Zhou; Li Shao; Lifu Zhao; Guoliang Lv; Xiaoping Pan; Anye Zhang; J. Li; Ning Zhou; Deying Chen; Lanjuan Li

Bioartificial livers may act as a promising therapy for fulminant hepatic failure (FHF) with better accessibility and less injury compared to orthotopic liver transplantation. This study aims to evaluate the efficacy and safety of a fluidized bed bioartificial liver (FBBAL) and to explore its therapeutic mechanisms based on metabolomics. FHF was induced by D-galactosamine. Eighteen hours later, pigs were treated with an FBBAL containing encapsulated primary porcine hepatocytes (B group), with a sham FBBAL (containing cell-free capsules, S group) or with only intensive care (C group) for 6 h. Serum samples were assayed using ultra-performance liquid chromatography-mass spectrometry. The difference in survival time (51.6 ± 7.9 h vs. 49.3 ± 6.6 h) and serum metabolome was negligible between the S and C groups, whereas FBBAL treatment significantly prolonged survival time (70.4 ± 11.5h, P < 0.01) and perturbed the serum metabolome, resulting in a marked decrease in phosphatidylcholines, lysophosphatidylcholines, sphingomyelinase, and fatty acids and an increase in conjugated bile acids. The FBBAL exhibits some liver functions and may exert its therapeutic effect by altering the serum metabolome of FHF pigs. Moreover, alginate–chitosan capsules have less influence on serum metabolites. Nevertheless, the alterations were not universally beneficial, revealing that much should be done to improve the FBBAL.


Metabolomics | 2012

Dynamic Patterns of serum metabolites in fulminant hepatic failure pigs

Pengcheng Zhou; J. Li; Li Shao; Guoliang Lv; Lifu Zhao; Haijun Huang; Anye Zhang; Xiaoping Pan; Wei Liu; Qing Xie; Deying Chen; Yongzheng Guo; Shaorui Hao; Wei Xu; Lanjuan Li

Fulminant hepatic failure (FHF) is still an intractable disease associated with serious metabolic disorder. Investigating the dynamic changes of serum metabolites during the development of FHF would facilitate revealing the pathogenesis and also promote its treatment. Therefore, this study characterized the dynamic metabonome of serum from FHF Pigs using ultra performance liquid chromatography–mass spectrometry. Based on multiple statistical analysis of the resulting dataset, three types of up-regulated and one type of down-regulated patterns were delineated. Each pattern demonstrated distinct trends at different stages during the whole process of FHF, implying the differential clinical significance of them. Specifically, aromatic amino acids (Pattern 1) and lysophosphatidylcholines (LPCs) (Pattern 4) might be good markers for evaluating the severity of FHF, while some conjugated bile acids, long chain acylcarnitines (Pattern 2) and Glycocholic acid (Pattern 3) could indicate liver injury in the early stage. Inspired from the PCA plot that the pathogenetic condition of FHF aggravated with sampling time, a linear discriminant analysis (LDA) model based on phenylalanine and LPC 18:1 were further constructed for evaluating the severity of FHF. The leave-one-out cross-validation accuracy of 91.67% for the training set and the prediction accuracy of 92.31% for the external validation set confirmed its excellent performance. In conclusion, findings obtained from the present study, including four types of Dynamic Patterns of serum metabolites during FHF development and an LDA model for evaluating the severity of FHF, will be of great help to the research and management of FHF in the future.


Transplantation Proceedings | 2014

Rapid Large-Scale Culturing of Microencapsulated Hepatocytes: A Promising Approach for Cell-Based Hepatic Support

Yunbo Chen; Chengbo Yu; Guoliang Lv; Hongcui Cao; Shigui Yang; Yi Min Zhang; Jun Yu; X. Pan; Lanjuan Li

INTRODUCTION The efficacy of any bioartificial liver device requires both rapid production and proper bioactivity of the cells for the bioreactor. The goal of this study was to observe the effect of spinner speed and cell density on the proliferation of microencapsulated immortalized human hepatocytes (HepLL) and human hepatoma (HepG2) cells. MATERIALS AND METHODS Alginate-chitosan microcapsulated HepG2 and HepLL cells were randomly divided into 2 groups, and each group was further divided into 8 subgroups according to embedded cell density and spinner speed. The growth, metabolism, and functions of the encapsulated cells in each group were evaluated. RESULTS In each group, the cell number, ammonium removal, albumin synthesis, and diazepam clearance increased significantly with the spinner speed, whereas embedded cell density had no impact. Albumin synthesis, removal of ammonium, and diazepam clearance were significantly higher in the microencapsulated HepLL groups than in HepG2 cells at any time point, without any significant difference in cell numbers. CONCLUSIONS Spinner culture significantly promoted microencapsulated HepLL and HepG2 cell bioactivity. Wrapped cells had optimal function on day 10 in rolling culture groups. These data show that HepLL cells would be a promising candidate for cell-based liver support therapy.


international conference of the ieee engineering in medicine and biology society | 2009

Construction of modular novel bioartificial liver support system

Jianfeng Liu; Tao Song; Wei Jiang; Yimin Zhang; Guoliang Lv; Lifu Zhao; Guanghao Zhang; Lanjuan Li

A modular novel bioartificial liver support system was designed and constructed in order to simplify tedious operation of artificial liver treatment and to improve the applicability in the system. The design ideas, structure composition, system function, and etc, were described in detail. In this system, the variety of the therapy modes could be conveniently connected by the interface of modular structure. Industrial control computer was used as the main control platform, and physical of control parameters such as pressure, pump speed, dissolved oxygen, temperature, and etc, were transmitted into computer, then according to the instruction, process of the treatment was accomplished by the executing units implemented by main control system. Touch screen of human-computer interface was adopted, which made the system better operational and more comfortable. The system has passed the spot function test, and all indexes can meet requirements for the clinical treatment requested. It has the character such as modular design, systematic distribution, building-block structure, and etc, which supports a great novel operation platform for artificial therapy.


Journal of Virological Methods | 2010

Expression of H5N1 influenza virus hemagglutinin protein fused with protein transduction domain in an alphavirus replicon system.

Shigui Yang; Jianer Wo; Minwei Li; Fen-fang Mi; Chengbo Yu; Guoliang Lv; Hongcui Cao; Haifeng Lu; Baohong Wang; Hanping Zhu; Lanjuan Li

Alphavirus replicons, in which structural protein genes are replaced by heterologous genes, express high levels of the heterologous proteins. On the basis of the potencies of replicons to self-replicate and express foreign proteins and the remarkable intercellular transport property of VP22, a novel alphavirus Semliki Forest virus (SFV) replicon system of VP22 fused with a model antigen, hemagglutinin (HA), of the human-avian H5N1 influenza virus, was explored in this study. Further, replicon particles expressing HA, VP22, and enhanced green fluorescent protein (EGFP) individually were used as controls. By flow cytometry based on the analysis of transfection efficiency, SFV-EGFP replicon particle titer was 1.13 x 10(7)transducing units (TU)/ml. The titers of SFV-HA, SFV-VP22 and SFV-VP22-HA replicon particles, which were titrated by using SFV-EGFP replicon particles, were 1.42 x 10(7), 3.23 x 10(7), and 1.01 x 10(7)TU/ml, respectively. HA and VP22-HA expression was observed in SFV-HA- and SFV-VP22-HA-transfected BHK-21 cells, respectively. Immunofluorescence staining revealed that the fluorescence intensity in the SFV-VP22-HA-transfected BHK-21 cells was more than that in the SFV-HA-transfected BHK-21 cells. Both SFV-VP22-HA and SFV-HA replicon particles presented a promising approach for developing vaccines against human-avian influenza. VP22-HA fusion protein with similar trafficking properties may also enhance vaccine potency.


biomedical engineering and informatics | 2011

Development of a novel artificial liver support system

Lifu Zhao; Guoliang Lv; Yimin Zhang; Anye Zhang; J. Li; Pengcheng Zhou; Xiaoping Pan; Yuemei Chen; Xiaopeng Yu; Chengbo Yu; Wei Xu; Ying Yang; Lanjuan Li; Jiansheng Xu; Tao Song; Yang Yu

Background/introduction: Artificial liver support treatment is a promising alternative to liver transplantation. An ideal artificial liver support system (ALSS) should be a combination of a nonbiological liver (NBL) device and a bioreactor based bioartificial liver (BAL). Material and methods: A novel ALSS which can not only fulfill toxin-removal functions of NBL but also provide biotransformation and synthetic functions of BAL is constructed. The unique dual-chamber reservoir can improve the efficiency of material exchange. The funnel-shaped fluidized bed bioreactor can provide an ideal physiological environment for hepatocytes. Quick bubble handling function improves the security during treatment. The software design provides error correction function. Our control center is an industrial personal computer and most components are integrated via the RS485 buses. The whole control system consists of three parts: a pump drive module, a sensor network and a human-machine communication interface. To verify our design, we test the system on miniature pigs. Results: The system runs normally in all treatment modes and meets the clinical requirements. Functions of all components are verified. Conclusions: The system provides a reliable research platform for artificial liver support treatment.

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J. Li

Zhejiang University

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