The production technology for the first generation of biofuels has been made great achievements and some countries, such as USA, EU countries, and Brazil, have formed successful biofuel industrial chains. Cellulose ethanol, a representative of the second generation of biofuels and a kind of more promising alternative fuels, has not yet got enough key technological breakthroughs and its large scale commercial production still needs rather some time to realize. Now it is in the early stage of transition to the second generation of biofuels. Taking the development of second generation of biofuels as their national policies, many countries have developed long-term development plans and objectives, and provided a good policy environment and strong support for it. Thus institutes and enterprises are engaging and successing in addressing key issues relating to the development of biofuels. At the mean time, some important issues relating to the sustainable development of biofuels have attracted people's attention.

Biomass resources are strategically vital for the sustainable development of a country.The United States has recently made a variety of plans or roadmaps and taken significant actions in order to promote the R&D of biomass resources.These plans and actions for the key issues in the development of biomass resources were analysed.

BACKGROUND: Recently, little attention has been paid to how to induce and identify the functions of differentiated cells in the methods for embryonic stem (ES) cells differentiation into hepatocytes. Whether the differentiated cells express functional characteristics of hepatocytes should be one of the markers to identify the hepatic differentiation of ES cells.OBJECTIVE: To direct mouse embryonic stem cells in vitro differentiation into functional hepatocytes by introduction of murine cholestatic serum in hepatocyte growth factor (HGF)-induced system.DESIGN: A controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: The experiment was carried out in the Medical Research Center of the Second Affiliated Hospital of Sun Yat-sen University from October 2004 to February 2007. The mouse E14 ES cell line was kindly provided by the Stem Cell and Tissue Engineering Center of Sun Yat-sen University. Twenty male SD rats, aged 2 weeks, were purchased from the Experimental Animal Center of Sun Yat-sen University. All animal experimental procedures were abided by the rules of animal ethnics.METHODS: The SD rats were undergone common bile duct ligation to induce cholestasis. Ten days after the operation, the whole blood of rats was collected to prepare cholestatic serum. The ES cells were cultured using hanging-drop method for 5-7 days to develop embryonic bodies (EBs). The dissociated EBs cells were then induced hepatic differentiation with spontaneous system, HGF (20 μg/L) system and cholestatic serum (5%) plus HGF (20 μg/L) system, respectively.MAIN OUTCOME MEASURES: The cellular morphologic changes were observed using transverse microscopy dynamically. (2) The cell staining for albumin, α-fetoprotein, CK18/19, glycogen, indocyanine green (ICG) and fluorescein diacetate (FDA) was done after 4 weeks differentiation. (3) The hepatocyte-specific metabolic functions of synthesizing albumin, triacylglycerol and urea nitrogen were assayed at 3 days interval.RESULTS: (1) The differentiation of ES cells cultured in spontaneous system was uncontrolled and the cells could grow into a wide range of three-germ cells. The HGF could promote ES cells differentiation into endoderm and mesoderm (myocardium). But the differentiated cells only expressed low levels of hepatic specific functions in these two induced systems. (2) Under cholestatic serum plus HGF system, the ES cells could differentiate into polygonal cells with very uniform morphology which were positive in glycogen, ICG and FDA staining and showed higher capabilities of synthesizing albumin, triacylglycerol and urea nitrogen than the differentiated cells in the other systems (P＜0.05-0.01).CONCLUSION: The cholestatic serum, a mimic pathological microenvironment in vitro, could effectively promote ES cells-derived hepatocytes induced by HGF to express high level of liver-specific metabolism functions.

0.05).No teratoma was formed in the experimental group,while a large teratoma was observed in control group in 6 weeks post-transplantation.CONCLUSION:The ESC-derived hepatic stem cells are normally incorporated into mouse liver parenchymal structure,proliferate and differentiate further in vivo and possess some hepatic functions without forming teratomas.

Objective To investigate the echocardiographic features and clinical significance of prenatal diagnosis of total anomalous pulmonary venous connection (TAPVC). Methods Fetal echocardiographic images of 13 fetuses with TAPVC conifrmed by pathology or postnatal echocardiography were reviewed. Echocardiographic features and clinical signiifcance of prenatal diagnosis of TAPVC were summarized. Results Twelve fetuses with TAPVC were diagnosed prenatally by fetal echocardiography, including seven cases of supracardiac type, three cases of infracardiac type and two cases of intracardiac type. The common echocardiographic characteristics of 12 fetuses with TAPVC included slightly size discrepancy of left heart and right heart, large foramen ovale with increased shunting at the atrial level, increased distance between left atrium (LA) and descending aorta, absent insertions of pulmonary veins in the LA, presence of pulmonary venous conlfuence on the top of LA and dilatation of vessels where pulmonary venous conlfuence drained. One case was missed prenatally and intracardiac type TAPVC was diagnosed by postnatal echocardiography. Among the 13 cases, three were isolated and the other ten were all in association with other abnormalities. Conclusions There are fetal echocardiographic characteristics of TAPVC. Fetal echocardiography plays an important role in prenatal diagnosis of TAPVC.

BACKGROUND: In vitro differentiation of embryonic stem cells into hepatocytes has been successfully reported to a certain degree; however, whether embryonic stem cells are able to effectively enter hepatic plate of host after intrahepatic transplantation, whether embryonic stem cells can further differentiate into hepatocytes and express hepatocyte function, and risk factors for neoplastic formation are still unclear at present. OBJECTIVE: To study the intrahepatic transplantation of embryonic stem cells-derived hepatic stem cells in therapeutic liver repopulation models, and to investigate the liver tissue replacement, growth and differentiation in vivo, and neoplastic formation.DESIGN: Randomized controlled animal study.SETTING: Department of Pediatric Surgery, the Second Hospital affiliated to Sun Yat-sen University. MATERIALS: Twenty-four BALB/c mice, 6-8 weeks old, weighing 20-35 g, irrespective of gender, were provided by Guangzhou Experimental Animal Center. Embryonic stem cells-derived hepatic stem cells were differentiated from embryonic stem cells. E14 was provided by Stem cell Center of our hospital. METHODS: This study was performed at the Stem Cell Center, the Second Hospital affiliated to Sun Yat-sen University from July 2006 to June 2007. Twenty-four mice were randomly divided into a liver repopulation model + stem cell transplantation group (group A) and a liver resection + stem cell transplantation group (group B), with 12 mice in each group. Mice in the group A were intraperitoneally injected with 50 mg/kg retrorsine once every two weeks for totally twice. Four weeks after the second injection, about 70% liver was resected. And then, the embryonic stem cells-derived hepatic stem cells, labeled by 1×105 carboxy fluoresce in diacetate succinimidyl ester (CFDA-SE), were transplanted into mouse liver through portal vein. On the other hand, 70% liver of mice in the group B was resected and embryonic stem cells-derived hepatic stem cells were transplanted into mouse liver. MAIN OUTCOME MEASURES: The distribution, incorporation, and proliferation of transplanted cells were observed under fluorescent microscopy. Two weeks later, hepatic function was stained with albumin fluorescence immunoassay (double fluorescence staining) and assayed by level of serum albumin. Embryonic stem cells-derived hepatic stem cells were poured into liver of remedial liver regeneration mice, and undifferentiated embryonic stem cells were transplanted into subcutaneous tissue in axillary region as the controls to observe neoplastic formation in embryonic stem cells-derived hepatic stem cells. RESULTS: ① Growth of hepatic stem cells in recipient mice: One week after transplantation of CFDA-SE-labeled embryonic stem cells-derived hepatic stem cells, some scattered region was green under fluorescent microscopy. The area of green region increased apparently in 2 weeks, and cord-like structure could be observed. ② Liver function: Immunofluorescent staining of albumin (double fluorescence staining) demonstrated that labeled cells expressed positive albumin (yellow fluorescence) in liver tissue of recipient mice, but there was not significant difference in serum albumin level between group A and group B (P > 0.05). ③ Reliability of hepatic stem cell transplantation: Teratoma did not form over 6 months; however, transplantation of undifferentiated embryonic stem cells in the axillary region could cause formation of teratoma after 6 weeks. CONCLUSION: The transplantation of embryonic stem cells-derived hepatic stem cells in therapeutic liver repopulation model mice can effectively and further grow and differentiate, or even partially express hepatocyte function; in particular, the transplantation is safe.

Objective The article was to observe the clinical efficacy of the application of terbinafine and mizolastine in com -bined treatment of chronic eczema ( CE) with dermatophytes infection , so as to define the etiology role of dermatophytes in allergic dis-eases. Methods All subjects were randomly divided into experiment group and control group .The experiment group was treated with the combination of terbinafine and mizolastine , while the control group took mizolastine orally alone .EASI grading , recovery rate and effective rate were evaluated at 2, 3 week after the treatment and EASI grading and recurrence rate were evaluated at 4 weeks after the treatment. Results 79 patients had finished the experiment .Significant difference was found in the effective rates between two groups at 3 weeks after treatment (P<0.05).At 4 weeks after the treatment, EASI value and recurrence rate in experiment group were obviously lower than those in control group (P<0.05). Conclusion Good therapeutic effect has been achieved through the ap-plication of terbinafine and mizolastine in combined treatment of CE with dermatophytes infection , which implies dermatophytes plays an important role in the etiology of CE .

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.

Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P＜0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P＜0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.

Objective To investigate the effect of low frequency electric deep brain stimulation on amygdale kindling in rats.Methods The amygdale kinkling model of rats was established by operation on the brain.The effects of low frequency deep brain electric stimulation used alone or in combination with anti-epilepsy drugs were ob- served in terms of severity of seizure attack reflected by Racine's scale and afterdischarge duration recorded in electro- encephalogram.Results Fifteen minutes of low frequency electric stimulation at 1 Hz and 100 to 350?A effective- ly inhibited amygdale kindling as demonstrated by a significant decrease of afterdischarge duration,and decreased the severity of seizure attack (P