Objective To investigate the mechanism of HSP70 that inhibits myocardial cell apoptosis in sepsis.Methods Myocardial cells in primary culture were randomly divided into control group,normal serum group,sepsis serum group,transported empty vector group and transported HSP70 group.The myocardial cells in transported HSP70 group have been transported by pcDNA3.1-HSP70 for 36 hours.The myocardial cells in every group have been cultured by respective serum for 2 hours and dyed by Hoechst 33258,and then calculate the rate of myocardial cells apoptosis.Using Western-blot to investigate the effect of overexpression of HSP70 on Caspase-3,8,9's activation and Bid's cracking.Results The rate of myocardial cells apoptosis after dealing in transported HSP70 group [ ( 12.48 ± 2.39 ) ％,( 23.96 ± 3.12 ) ％,( 25.40 ± 3.96) ％ ] is lower than in sepsis serum group [ ( 28.66 ± 2.24 ) ％,( 55.76 ± 5.69 ) ％,( 46.89±8.74)％,t =5.856,5.932,6.027,P ＜0.01,n =3] and lower than in transported empty vector group [(34.25±3.42)％,(50.71±6.38)％,(47.62+5.74)％,t =5.876,5.903,6.122,P ＜0.01,n =3],is higher than in control group,and in normal serum group(3.13％ ～ 6.75％ ,t =6.324,6.578,6.137,5.987,6.032,6.871,P ＜ 0.01,n =3 ).When Caspase-3,8,9 activating,the gray-scale of P11,P20 and P10 in transported HSP70 group( 12.5276 ± 2.1247,9.3481 ± 4.5423,16.1349 ± 6.0641 ) is lighter than that in sepsis serum group ( 27.1324 ± 2.1564,25.5643 ± 4.3018,36.5647 ± 6.7135,t =5.856,5.902,5.891,P ＜ 0.01,n =3 ) and lighter than in transported empty vector group (28.0314 ±2.0367,25.6413 ±4.1356,34.5648 ±5.9473,t =3.861,3.933,4.281,P ＜0.05,n =3),is deeper than in control group(8.0324 ± 1.5234,5.1246 ± 1.3274,2.0314 ±0.6423,t =3.286,3.867,4.031,P＜0.05,n =3) and in normal serum group(8.5649 ± 1.2136,6.0324 ± 1.0214,3.2146 ±0.1325,t =5.898,5.969,6.879,P ＜0.01,n =3).The gray-scale of tBid in transported HSP70 group( 12.0316 ±2.3641 ) is lighter than in sepsis serum group(27.0536 ± 5.3214),t =3.274 ( P ＜ 0.05,n =3 ) and lighter than in transported empty vector group(27.1034 ± 3.6741,t =3.301,P ＜ 0.05,n =3 ),is deeper than in control group ( 6.0347 ± 2.1304,t =5.924,P ＜ 0.01,n =3 ) and in normal serum group ( 7.3121± 1.3021,t =5.871,P ＜ 0.01,n =3 ).Conclusions HSP70 inhibit myocardial cells apoptosis in sepsis by intervened the death receptor pathway and mitochondrial pathway.

Objective To validate genetic suppression of metastastic capability of highly metastastic melanoma cells by endostatin transfection.Method pcDNA3.1-Endo eukaryotic expression vector contained insulin signal peptide sequence was transfected into highly metastatic mice melanoma cell strain B 16.The expression of endostain was detected by RT-PCR and Western blot experiment,melanoma cells were determined with adhere experiment,in vitro invasion and migration experiment and pulmonary metastasis experiment on C57BL/6 mice.Result Endostatin can obviously inhibit the capability of adherence,in vitro invasion and migration and pulmonary metastasis of melanoma cells.Among them,adhere inhibition ratio was 67.3%,in vitro invasion inhibition ratio was 48.4%,cell migration inhibition ratiowas 52.1%and pulmonary metastasis inhibition ratio was 67.3%.Conclusion Endostatin transfection can obviously inhibit the highly metastatie capability of melanoma cells.

Objective To investigate the efficacy and safety of intravenous itraconazole in different antifungal strategies for hematologic diseases patients with invasive fungal disease. Methods The efficacy and safety of intravenous itraconazole injection in the treatment of 160 hematologic diseases patients with invasive fungal disease, including the related factors were retrospectively analysed. Results The total efficacy rate of itraconazole was 58.12 %(93/160). The response rates in therapy for undefined patients without any evidence of patients, diagnostic-driven therapy for possible IFD patients, targeted therapy for proven IFD patients were 65.82 %(52/79), 53.57 %(30/56) and 44.00 %(11/25), respectively (P=0.054). The incidence rate of itraconazole-related adverse effect was 8.13 % (13/160), and the main adverse reaction was liver impairment. Multiple-factor analysis showed that the efficacy of itraconazole for the treatment of hematologic diseases patients with invasive fungal disease was not associated with age, medical history, agranulocytosis, and initial treatment. Conclusion Itraconazole itraconazole is effective and safe in the treatment of fungal therapy for patients with hematologic diseases.

Aim To explore the mechanisms of the apoptosis induction and the effects of adhesion suppression of Zhiling capsule (ZLJN) in small cell lung cancer cell line NCI-H446.Methods According to the different components of ZLJN,NCI-H446 cells were treated with traditional Chinese medicine,western medicine and ZLJN composite groups.Apoptotic cells were tested by light microscopy,Hochest33258 staining method.The mRNA and protein expressions of bcl-2,bax and hTERT were analyzed by RT-PCR and Western blot respectively.The expressions of CD44 were detected by flow cytometry.Results After NCI-H446 cells were treated with different drug groups,The morphological changes of apoptotic cells were found by light microscopy and Hochest33258 staining method.The mRNA and protein expressions of bcl-2 were down-regulated while the expressions of bax were up-regulated compared to the control groups(P

Aim To investigate the effects of Zhiling capsule (ZLJN) on the proliferation inhibition and apoptosis induction in K562 cell line.Methods According to the different components of ZLJN,K562 cells were treated respectively with tradtional Chinese medicine,Western medicine and ZLJN compound groups.The cell viability and colony formation were observed by MTT assay and colony formation assay respectively.Apoptotic cells were detected by Annexin V-FITC/PI staining and DNA fragmentation assay.Caspase-3 activity was detected by flow cytometry,and pro-caspase-3 was detected by Western blot.Results Treated with drug,K562 cell growth and cell colony formation were significantly inhibited.Apoptosis occurring in the early stage was identified by Annexin V-FITC/PI staining.Typical DNA ladder was seen from gel electrophoresis and apparent apoptotic peaks were observed by flow cytometer.The level of caspase-3 activity increased after the treatment,while the level of pro-caspase-3 decreased.Conclusion ZLJN can efficiently inhibit proliferation and induce apoptosis in K562 cells,which may be related with the up-regulation of caspase-3 activity.

Aim To observe the adhesive process of the human gastric cancer cell line MKN1,and study the expression of integrin ?1;to investigate the mechanism of the inhibition of the adhesion process of MKN1 cells by destran sulfate(DS).Methods The MKN1 cells were cultured with DS or PBS,then stained with immunofluorescent cytochemistry and observed in fixed or living conditions with confocal laser scanning microscope.RT-PCR was used to analyze the cDNA expression of MKN1 cells.Results MKN1 cells adhered to culture dishes by the process of forming filopodia,changed into a flat shape,and then adhered to other cells to form a cell-monolayer.Integrin ?1 was intensively expressed in the cell membrane,where integrin ?1 formed clusters.DS inhibted the expression of integrin ?1 in cell membrane,and decreased the area of integrin ?1 clusters.DS-treated cells also tended to maintain a round shape by contracting the filopodia.In DS-containing culture dishes,some cells kept floating 4 hours after seeding.DS decreased the level of the cDNA expression of the adhered cells to 74% and of the floating cells to 38% of that of the cells in un-treated group,respectively.Conclusion The inhibition of the adhesion of MKN1 cells by DS was related to the suppression of the expression of integrin ?1.

Objective To study the influence of let-7b on cell proliferation and aerobic glycolysis of human melanoma cell A375.Methods Transfect A375 cell line with hsa-let-7b oligonucleotide or antisense.Glucose and lactate in medium were determined by spectrophotometry at 24 h and 48 h time point after transfection.The cell proliferation was determined by methylthiazol tetrazolium (MTT) assay.Results Over expression of let-7b in melanoma cell reduced cell proliferation notably,compared to the other groups by MTT(P ＜0.05).However,the glucose consumption and lactate production differences were not observed during 24 h or 48 h ( P ＞ 0.05 ),the blank control group transformed about 57％ and 43％ glucose to lactate during 24 h and 48 h.Conclusions Melanoma cell line A375 has notably aerobic glycolysis hallmark,let-7b could inhibit proliferation of melanoma cell line A375,but it may has no influence on glucose metabolism.

Objective To confirm whether or not let-7b and miR-199a were significantly associated with malignant melanoma growth and proliferation. Methods An over -expression plasmid and an inhibitor, which targeted on let-7b and miR-199a, was constructed. B16F10 cells were divided into seven groups: control group, let-7b plasmid group, miR-199a plasmid group, empty plasmid group, let-7b inhibitor group, miR-199a inhibitor group, inhibitor control group. Foreign gene was transfected into B16F10 cells, let-7b and miR-199a expression were validated from RNA level, protein level and cell level. Results The relative let-7b or miR-199a gene expression of the let-7b plasmid group (3.8776±0.1372)and miR-199a plasmid group (2.8660±0.2821)were significantly higher than control group (P＜0.05), the relative let-7b or miR-199a gene expression of the let-7b inhibitor group (0.2057±0.0263) and miR-199a inhibitor group(0.2656±0.0253) were significantly lower than control group(P＜0.05). The cyclinD1 expression of the let-7b plasmid group(2.023±0.315) and let-7b inhibitor group (1.857±0.377) were significantly higher than control group (0.997±0.041) (P＜0.05), whereas, the Met expression of themiR-199a plasmid group (5.19±0.309) and miR-199a inhibitor group (4.87±0.044) were significantly higher than control group (2.2±0.198) (P＜0.05). The let-7b plasmid group and miR-199a plasmid group B16F10 cell growth rate were slower than control group, especially on the third day after transfection, the growth rate gradually dropped to the lowest value (P＜0.05). In addition, the apoptosis rates of the let-7b plasmid group and miR-199a plasmid group reach to (11.8±1.19)% and (11.3±1.59)%,which were significantly higher than control group (P＜0.05). Conclusions let-7b and miR-199a may be a negative regulator on the B16F10 cell growth and proliferation.

MicroRNAs have been identified as a new class of regulatory molecules that affect many biological functions by interferring the target gene expressions. Latest studies demonstrate that microRNAs can influence many pivotal bio-processes and deeply involve in the metabolism of glucose, lipid and amino acid and biological oxidation. For glucose metabolism, microRNAs are related to insulin secretion, insulin sensitivity, glucose uptake, glycolysis, oxidation and mitochondrial function. For lipid matebolism, microRNAs can regulate the target genes related to lipid biosynthesis, catabolism and transportation. MicroRNAs can influence glutamine catabolism.
Animals ; Glucose ; metabolism ; Glutamine ; metabolism ; Humans ; Insulin ; metabolism ; Insulin Secretion ; Lipid Metabolism ; physiology ; Metabolism ; physiology ; MicroRNAs ; physiology

Objective To set up and apply a standardized communication system of adolescents and young adults (AYAs) cancer patients,in order to improve AYAs cancer patients' psychological distress and other negative emotions,as well as promote social support and quality of life of patients.Methods A AYAs cancer patients standardized communication system,suitable for China's national conditions,was preliminarily built.Using randomized controlled trials,a total of 171 subjects,selected from 486 cases of AYAs cancer patients,in the Third Xiangya Hospital of Central South University and Hunan Cancer Hospital from August to September in 2016,were intervened with a set of standardized communication system.The Mental Distress Thermometer (DT),Hospital Anxiety and Depression Scale (HADS),Social Support Rating Scale (SSRS) and Concise Health Status Questionnaire (SF-36) were used as evaluation indexes to observe the psychological distress,emotion,social support and quality of life of the three groups of subjects before intervention,immediately after intervention,1 month after intervention and 3 months after intervention.Results At the follow-up of 3 months after intervention,38 cases were lost,14 cases in communication group (final n =43),11 cases in music therapy group (final n =46),and 13 cases in routine group (final n =44)..There was no significant difference in the evaluation indexes between the communication group,music treatment group and the routine group before intervention (P ＞ 0.05).There were statistically significant differences in scores of psychological pain,anxiety and depression,and social support in the 3 groups,before and immediately after intervention,1 month and 3 months after intervention (P ＜ 0.05).The scores of psychological pain,anxiety and depression,social support and quality of life in the communication group 1 month after intervention were statistically significant compared with those in the music group and the routine group (P ＜ 0.05).Conclusions Compared to music therapy and regular care,the standardized communication system has significant effect on improving the psychological distress of AYAs with cancer,and can also improve their social support level and quality of life.The clinical validation of the standardized communication system can provide reference for psychological rehabilitation of cancer survivors.