BACKGROUND:In recent years, embryonic hepatic stem cel s have attracted more attention, but there are few reports on the potential of embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s as wel as the related differentiation conditions. OBJECTIVE:To investigate the moderate condition to induce mice embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s in vitro with chemical reagents. METHODS:Dimethylsulfoxide in combination with 5-azacytidine with different concentrations and time were used to induce the embryonic hepatic stem cel s of 13.5 days mice and to observe the differentiation effect. RESULTS AND CONCLUSION:Under in vitro conditions, 0.8%dimethylsulfoxide+5μmol/L 5-azacytidine could induce the mouse embryonic hepatic stem cel s to express the specific markers of myocardial cel s, while increasing the concentration of the inducer and extending the induction time could not improve the induction efficacy.

10.3969/j.issn.2095-4344.2013.23.014

BACKGROUND: Fetal liver cells can have stronger abilities to proliferation and differentiation and lower immunogenicity compared to bone marrow stem cells. However, there are few studies on direct isolation and culture of embryonic hepatic stem cells (EHSCs). OBJECTIVE: To isolate and cultivate EHSCs in vitro and to identify their biological features. DESIGN, TIME AND SETTING: The cytology in vitro controlled study was performed at the Chongqing Key Laboratory of Neurology from March to June 2008. MATERIALS: A total of 9 SPF Kunming fetal mice aged 13.5 days were obtained from Animal Experimental Center of Chongqing Medical University. METHODS: Collagenase + EDTA digestion and differential adherence method were used to isolate EHSCs, which were then incubated at 2?108 /L. Cells were digested and passaged when 80%-90% cells were confluent. Using streptavidin-biotin-peroxidase complex technique, adhered cells following 5 days of incubation were labeled with various EHSC surface marker. MAIN OUTCOME MEASURES: Morphology, passage, amplification and surface marker surface of EHSCs were measured. RESULTS: The isolated EHSCs adhered to the culture plastic and presented pykno-round cells and distinct borderline 24 hours after cultivation in vitro. Cells grew spindle-shaped 3 days. After 7 days they grew like epithelium. Cell amplified speed following passage did not have significant changes. Cells still presented epithelium-like shape at the passage 5. The adhered cells at day 5 following primary incubation were positively for human stem cell factor receptor and alpha fetoprotein, and negatively for albumin and cytokeratin 19. CONCLUSION: EHSCs were positively for human stem cell factor receptor and alpha fetoprotein, and negatively for albumin and cytokeratin 19 in early primary culture. This indicated that the cultivated cells are proved to be primordial progenitor cells and still in undifferentiated early phase.

Objective: To investigate the antitumor metabolites of fungal mutants 2-2-3 and PDN-f-2, the two bioactive mutants of Penicillium purpurogenum G59 that do not produce antitumor metabolites. Methods: Bioactive metabolites newly produced by the mutants were isolated by a bioassay-guided separation procedure using iquid-liquid extraction, column chromatography and recrystallization methods through direct comparison with the sample from P. purpurogenum G59. The compounds obtained were identified by spectroscopic methods. The antitumor activity was assayed by the MTT method using K562 cells. Results: Two bioactive metabolites 1 and 2 were isolated from the fermentation products of 2-2-3 and PDN-f-2, respectively, and identified as ergone (1) and citrinin (2). Compounds 1 and 2 inhibited the proliferation of K562 cells with the IC50 values of 7. 4 and 48. 0 μg/ml, respectively. Conclusion: Compounds 1 and 2 are the antitumor metabolites newly produced by he mutants 2-2-3 and PDN-f-2, respectively, and have not been found in the metabolites of P. purpurogenum so far. It is revealed from the present result that the alteration of secondary metabolism of wild-type fungal strains without bioactivity for obtaining bioactive metabolite-producing mutants may become a new route to expand the source of new fungal strains for drug screening.

Humans ; Purpura, Thrombotic Thrombocytopenic ; diagnosis ; therapy

The liver is an important organ of the body,with an extraordinary capacity for responding to physical or chemical injuries by regenerating. The mechanisms behind liver regeneration are very complicated as hundreds of substances are involved.However,most of them are not liver-specific,such as HGF,EGF etc. We recently isolated a pure protein with hepatic stimulatory activity from the extract of a weanling calf liver and named its hepatopoietin Cn (HPPCn).This paper,with reference to our own work, mainly reviews the development and bioactivity about hepatic stimulator substance(HSS),augmenter of liver regeneration(ALR),hepassocin(HPS) and HPPCn,which are liver-specific biologically active peptides.

We predicted the anti-ulcerative colitis (UC) mechanism of Fructus Amomi based on network pharmacology. The anti-UC activity of Fructus Amomi were investigated by in vivo animal experiment, and the active components of Fructus Amomi were obtained through TCMSP, PubChem database and literature research. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of Institute of Materia Medica of Chinese Academy of Medical Sciences. The potential targets of the active components and UC were predicted by SwissTargetPrediction, GeneCards and TTD databases. The protein-protein interaction (PPI) network was constructed by String database and Cytoscape software was used to construct a visual network of active component-disease target and perform topological analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using Metascape platform. The molecular docking of key components and core targets was carried out by Sybyl X software. We screened out a total of 12 active components and 189 disease-component overlapping targets. Enrichment analyses obtained 227 related GO items and 168 signaling pathways. According to the results of molecular docking, most active components of Fructus Amomi showed good affinity with the JAKs receptor family. Furthermore, Western blot results verified that Fructus Amomi could effectively inhibit JAK/STAT signaling pathway, indicating that Fructus Amomi might exert the anti-UC activity by regulating JAK/STAT signaling pathway.

Objective To explore the relationship of insulin resistance with serum 25-hydroxyvitamin D levels and arterial stiffness in elderly patients.Methods A total of 162 elderly inpatients (aged 60 years or over) were recruited from 2012 to 2014.Levels of fasting serum insulin,fasting serum glucose,creatinine and vitamin D were determined.Insulin resistance (IR) was evaluated by the homeostasis model assessment of insulin resistance (HOMA-IR).The patients were divided into the following three groups according to HOMA-IR:the low IR group with HOMA-IR less than 2 (n=78),the median IR group with HOMA-IR between 2 and 6 (n=43),and the high IR group with HOMA-IR greater than 6 (n=41).Arterial stiffness was assessed by carotid-femoral pulse wave velocity (cfPWV).Arterial stiffness and serum vitamin D levels were compared among the three groups.Results cfPWV was increased and the serum 25-hydroxyvitamin D level was decreased in the high insulin resistance group compared with the low insulin resistance group[(13.2± 5.7) μg/L vs.(17.8±6.2) μg/L,(14.3±5.2) m/s vs.(11.9±3.0) m/s].Multiple liner regression analysis showed that IR was negatively correlated with the serum 25 hydroxyvitamin D level (r=-0.63,P＜0.05) and positively correlated with arterial stiffness (r=0.45,P＜0.05) after adjustment for age,sex and other confounders.Conclusions Elderly patients with high insulin resistance may have lower levels of serum vitamin D and higher arterial stiffness.

In order to realize the sustained education concept in clinical medical English teaching,several measures were taken in the first clinical medical college of Nanjing University of Traditional Chinese Medicine,such as training the teaching staff,using original textbooks and redesigning the curriculum.Particularly the tutorial system was introduced to the education frame.The teaching and research section of clinical medical English explored the new teaching routes for postgraduates in traditional Chinese medicine university.

Objective To explore the efficacy and security of helical tomotherapy (radical radiotherapy) combined with targeted therapy for stage Ⅳ non-small cell lung cancer (NSCLC).Methods The data of 19 patients with stage Ⅳ NSCLC were retrospectively analyzed.The efficacy and security were observed by SPSS 20.0 statistical software.Results Among 19 patients, there were 6 cases complete remission (CR), 8 cases partial remission (PR), 4 cases stable disease (SD), 2 cases progressive disease (PD).The response (CR+PR) rate was 73.7 ％ (14/19).The 1-and 2-year local control rates were 73.7 ％, 47.4 ％,respectively.The 1-year progression-free survival rate was 31.6 ％.The median survival time was 13 months, and the 1-and 2-year overall survival rates were 52.6 ％, 26.3 ％, respectively.The median survival time of females, non-smokers or targeted therapy synchronous radiotherapy was longer than that of male, smokers or targeted therapy sequential radiotherapy, respectively (21 months vs 8 months, P =0.014;21 months vs 6 months, P =0.007;18 months vs 6 months, P =0.026).There were no significant differences of age, number of metastatical organs or targeted drug categories between two groups (P ＞ 0.05).All patients could tolerate the adverse reactions without treatment-related death.Conclusion The effect of helical tomotherapy combined with targeted therapy for stage Ⅳ NSCLC is desirable.