Objective To investigate the intervening effects of Ganoderma spore on the decrease of cell proliferation and neuronal survival in the hippocampus of fetal and postnatal rats induced with gestational hypertension. Methods Fourty SD pregnant rats were divided into four groups including the control group,Nw-nitro-L-arginine methylester(L-NAME)+distilled water(DW) group,L-NAME+L-Arginine group and L-NAME+Ganoderma spore(GS) group.The hippocampal tissue of the brain was detected by immunohistochemistry,Western blotting,RT-PCR,flow cytometry and electron microscopy. Results After the application of L-NAME,the expressions of hypoxia inducing factor-1?(HIF-1?) and vascular endothelial growth factor(VEGF) were increased at the hippocampus of E21 brain and continued up to P30 brain.The microvessel density of E21 hippocampus was increased and the structural abnormalities of blood capillary at P30 hippocampus were showed.The cell proliferation was decreased at E21 hippocampus and so was the neuronal number at P30 hippocampus.With the administration of Ganoderma spore,HIF-1? and VEGF were down-regulated at E21 hippocampus and were not detected at P30 hippocampus.The microvessel density of E21 hippocampus reached a normal level and the blood capillary ultrastructure of P30 hippocampus was restored.The cell proliferation of E21 hippocampus and neuronal number of P30 hippocampus were recuperatively increased.Conclusion Ganoderma spore may prevent the decrease of cell proliferation and neuronal survival in the hippocampus of fetal and postnatal rats induced with gestational hypertension.

12 months, baseline serum HBV DNA ≥104copies/ml) were prospectively followed. Virological and serologic responses and laboratory data of liver and renal function of each patient enrolled were monitored every three months. Entecavir resistant mutations were detected by direct sequencing of HBV reverse transcriptase region which was amplified by semi-nested polymerase chain reaction (PCR). HBV sequences of different genotypes obtained from GenBank were aligned to identify entecavir drug resistant substitutions. Results Of the 41 lamivudine refractory CHB patients who met the inclusion criteria, 16 patients (39%) achieved the point of undetectability in HBV DNA level (

Objective To investigate the influence of aging on motor function,binding activity and protein expression level of striatum dopamine transporter(DAT)of rats. Methods Rolling-bar test was performed to assess motor function.Western blot and 131I-FP-β-CIT up-take ratio were used to evaluate DAT protein 1evel and striatum DAT binding activity respectively. Results There were an age-related decline of rolling-bar latency and striatum 131I-FP-β-CIT up-take ratio in rats older than6 months.There was a significant difference between 6-month-old rats and older rats(12-month-old rats,16-month-old rats,20-month-old rats),and rolling-bar latency was correlated with striatum 131I-FP-β-CIT up-take ratio(r=0.656,P＜0.01).There was no change in DAT protein 1evel during aging process. Conclusions The age-related decline of motor function of rats was correlated with the decreasing of striatum DAT binding activity.The synaptic membrane expression of striatum DAT may decrease in aged rats.

OBJECTIVE: To investigate the effects of Ganoderma spores on mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension. METHODS: Nitric oxide synthase (NOS) inhibitor Nw-nitro-L-arginine methylester (L-NAME) was intraperitoneally injected into pregnant rats to induce gestational hypertension, and Ganoderma spores were administered orally. The effects of Ganoderma spores on levels of mitochondria-related molecular substances in hippocampus of young rats birthed by the rats with gestational hypertension were evaluated with immunoradiometric assay of cAMP, RT-PCR analysis of related genes, and detection of enzyme activity. RESULTS: In hippocampus of the new-born rats birthed by rats with gestational hypertension, the cAMP level, mitochondrial DNA (mtDNA) level and adenosine triphosphatase (ATPase) activity were decreased, and the expression level of peroxisome proliferator activated receptor gamma coactivator 1 alpha (pgc1 alpha) was unchanged compared to the normal control group. The cAMP level, mtDNA level, ATPase activity and pgc1 alpha expression level in hippocampus of 30-day post-natal rats were lower than those of the rats in normal control group. After oral administration of Ganoderma spores, the cAMP and mtDNA levels in hippocampus of the new-born rats and 30-day post-natal rats recovered almost to the levels of normal control rats, and the ATPase activity and pgc1 alpha expression level were also increased significantly. CONCLUSION: Ganoderma spores may regulate the levels of mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension.

OBJECTIVE: To detect some proteins associated with the effect of ganoderma lucidium spores (GASP) on promoting the survival and axon regeneration of injured spinal motor neurons in rats. METHODS: The rats were divided into normal control group, untreated group and GASP-treated group, and the rats in the last two groups received ventral root avulsion. GASP preparation was fed to the rats in the GASP-treated group for 14 days. The gray matter tissues of the lumbar spinal were sampled from rats in each group after 14 days following ventral root avulsion, and the extracted proteins from these tissues were detected by using 2-dimensional electrophoresis. Matrix assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) was utilized to identify the differentially expressed proteins among these three groups. RESULTS: There were six kinds of proteins differentially expressed among the three groups, which were collapsin response mediator protein 2 (CRMP-2), F-actin capping protein beta subunit (FCP-beta), isocitrate dehydrogenase [NAD] subunit beta (IDH-beta), ATPase, glutamate oxaloacetate transaminase-1 (GOT1) and M2 pyruvate kinase (M2-PK). The expression levels of CRMP-2, IDH-beta, ATPase and GOT1 were higher in the GASP-treated group than those in the untreated group, while the expression levels of FCP-beta and M2-PK were lower than those in the untreated group. CONCLUSION: GASP maybe promotes the survival and axon regeneration of injured spinal motor neurons in rats by virtue of up- or down-regulating the expression levels of the proteins mentioned above.

Objective To observe the effect of mutant α-synuclein(A30P)in autophagic programmed cell death by transfected PC12 cells and explore its probable role and pathway in PD.Methods The definite PC12 cells which were transfected mutant α-synuclein(A30P)were constructed at first and MPP+,Rapamycin and Wortmanin were administrated to transfected PC12 cells with mutant α-synuclein. Not only the proliferative activity of cells was detected with MTT method but also the ultrastructttre changes of cells and expression of α-synuclein in different circumstance were observed by transmission electron microscopy(TEM),Western Blot and the level of SOD.Results (1)The expression of α-synuclein in groups A30P+Wortmannin and A30P+MPP+was higher than that in group A30P(P＜0.01), particularly.there was more significant expression of α-synuclein in group A30P+Wortmannin.The expression of α-synuclein in group A30P+Rapamycin was weaker than that in group A30P(P＜0.01); (2)The results showed that the SOD level(group A30P+MPP+:3 h:97.49±13.8;12 h:102.7±12.7; 24 h:101.5±11.8;48 h:104.3±12.4)was significantly decreased at various time points after MPP+ treatment compared that of group A30P(t=3.7721,P=0.0017).SOD level gradually increased in A30P +Rapamycin 12 h and showed significant difference at 24 h(121.2±13.0),48 h(124.3±14.1)and 72 h(127.7±13.7)after drug treatment compared with that in group A30P+Wortmannin(t:2.9746, P=0.0083);(3)Mutant α-synuclein(A30P)leading to PC12 cells death by means of autophagy involved α-synuclein accumulation,membrane lipid oxidation,and loss of plasma membrane integrity.Mutant α- synuclein(A30P)mediated the toxicity of MPP+.Rapamycin,an inducer of autophagy,reduced the aggregation of α-synuclein in transfected cells.Meanwhile,Wortmanin,an inhibitor of autophagy,promoted the aggregation of α-synuclein in transfected cells and induced cells to die.Conclusions The abnormal aggregation of α-synuclein induces autophagic programmed cell death in PC12 cells and mutant α-synuclein (A30P)mediates the toxicity of MPP+.Meanwhile,Rapamycin may reduce the aggregation of α-synuclein in transfeeted cells by activation of autophagic pathway.

BACKGROUND:The studies have shown that the mesenchymal stem cel s derived from bone marrow and umbilical cord can be continuously cultured in vitro, and maintain the characteristics of stem cel s. The mesenchymal stem cel s can differentiate into hepatocyte-like cel s after“cocktail”induction by various cytokines. OBJECTIVE:To further identify whether umbilical cord-derived mesenchymal stem cel s in vitro co-cultured with normal hepatocytes can differentiate into hepatocyte-like cel s, and to investigate the differentiation method. METHODS:Mesenchymal stem cel s were isolated from human umbilical cord with adherent method, and the surface markers of umbilical cord-derived mesenchymal stem cel s were detected with flow cytometry. The umbilical cord-derived mesenchymal stem cel s were co-cultured with liver LO2 cel s without adding exogenous inducers. The expressions of alpha-fetoprotein, albumin and human cytokeratin 19 mRNA of hepatocyte specific markers were detected with reverse transcription PCR at 7, 14 and 21 days after culture, and periodic acid-Schiff staining was used to identify the functions. RESULTS AND CONCLUSION:Mesenchymal stem cel s could isolated from human umbilical cord successful y, showing fibroblastic morphology and adherent cel characterization. Among these cel s, 96.02%cel s were CD29 positive cel s and 96.6%cel s were CD105 positive cel s. The percentage of CD34 negative cel s was 99.65%. The percentage of CD105+CD29+double positive cel s was 94.84%. The mRNA of alpha-fetoprotein was found on the 7th day after co-cultured with LO2 cel s, and the mRNA of albumin and human cytokeratin 19 were found on the 14th day. After co-cultured for 21 days, the alpha-fetoprotein mRNA could not be observed in the co-culture group. The expressions of albumin and human cytokeratin 19 were increased at 14 days. After co-cultured for 21 days, the glycogen staining was positive. Umbilical cord-derived mesenchymal stem cel s can differentiate into hepatocyte-like cel s after co-cultured with normal hepatocytes.

OBJECTIVETo observe the clinical characteristics and short-term survival of patients with splenomegaly and acute-on-chronic liver failure related to chronic HBV infection.

METHODSElectronic medical records of patients with acute-on-chronic liver failure were collected to analyze the clinical parameters and 4-week survival of patients with or without splenomegaly.

RESULTSOf the 149 patients enrolled, the overall 28-day mortality rate was 48.3%, which was lower in patients with enlarged spleen than those without (34.2% vs 54.1%, P=0.034). Compared with patients without splenomegaly, patients with splenomegaly had lower platelet counts (P=0.001), lower ALT levels (P=0.005) and lower PT-INR (P=0.010). Although the occurrence of hepatic encephalopathy was comparable between patients with or without splenomegaly, severe conditions were more frequent in those without splenomegaly. Hepatic encephalopathy grades, serum creatinine levels, neutrophil percentages over 70%, PT-INR and splenomegaly were independent factors associated with the 28-day survival, and this novel model was superior to model of end-stage of liver disease in predicting the 4-week survival (P=0.017).

CONCLUSIONPatients with splenomegaly that evolves into acute-on-chronic liver failure have unique clinical characteristics and further clinical observations are warranted.

Acute-On-Chronic Liver Failure ; mortality ; physiopathology ; Chronic Disease ; Hepatic Encephalopathy ; physiopathology ; Humans ; Splenomegaly ; mortality ; physiopathology

Transcatheter arterial chemoembolization (TACE) is recommended by domestic and international guidelines for the treatment of patients with unresectable hepatocellular carcinoma (uHCC), and it is one of the most common treatment methods for patients with uHCC. The chemotherapy drugs commonly used in TACE for HCC include epirubicin, cisplatin, and fluorouracil, while it is still unclear which chemotherapy drug has a better clinical effect. This article summarizes the studies of different TACE regimens using different chemotherapy drugs in the treatment of patients with uHCC in the recent five years. TACE combined with sorafenib can significantly improve the survival of patients with advanced HCC and has been recommended for the treatment of such patients by Chinese Society of Clinical Oncology guidelines, and the efficacy of TACE combined with other tyrosine kinase inhibitors (TKI) has become a research hotspot. Studies have shown that compared with TACE combined with sorafenib in the treatment of patients with advanced HCC, TACE combined with lenvatinib can achieve a significantly longer progression-free survival time and a tendency of increase in median overall survival time. However, due to the variation of target receptors or downstream signals, resistance to molecular-targeted agents is still a challenging problem. TKI combined with immune checkpoint inhibitors may be a promising strategy for the treatment of patients with uHCC. Some studies suggest that triple therapy using TACE combined with TKIs and anti-PD-1/PD-L1 monoclonal antibody has better efficacy in improving the survival of patients with uHCC. This article reviews the studies of the efficacy and safety of TACE combined with targeted agents and TACE combined with anti-PD-1/PD-L1 monoclonal antibody in the treatment of patients with uHCC in the recent five years.