Objective To investigate the p53,Bax,bcl-2 gene in NaAsO2-induced human embryonic lung fibroblasts(HELF)apoptosis.Methods HELF was divided into HELF cells transfected with p53 plasmid(p53 group),HELF cells transfected with PC plasmid(PC group)and normal cultured HELF cells(normal group).The mRNA expression of p53,Bax and bcl-2 gene was detected by real-time PCR,the protein expression of p53,Bax and bcl-2 was assessed by immunohistochemical SABC and the cell apoptosis of HELF was detected by flow cytometry(FCM),in a 6-well plate and cultured for 48 hours,which was exposed to different doses(0,3,9,15mmol/L)NaAsO2 for 24 hours.Results The p53 gene mRNA expression level of p53 group(0.51±0.29)was lower than that of the normal group and PC group [ (1.00 ± 0.20), (1.32 ± 0.26), all P < 0.05 ]. The p53 protein expression level of p53 group(4.10 ± 1.20) was lower than the PC group and normal group[ (8.00 ± 1.63), (7.90 ± 1.79), allP < 0.05]. In p53 group, PC group, normal group exposed to 0,3,9,15 mmol/L NaAsO2 doses, the apoptotic rate [(0.57 ± 0.28)%, (22.91 ± 4.86)%, (40.05 ± 3.93)%, (44.87 ± 3.58)%; (0.65 ± 0.24)%, (14.09 ± 3.49)%,(20.31 ± 3.66)%, (32.42 ± 3.63)%; (0.56 ± 0.25)%, (12.14 ± 3.70)%, (19.61 ± 3.63)%, (30.43 ± 2.83)%], Bax mRNA expression level[(12.73 ± 3.96), (25.12 ± 6.42), (104.96 ± 26.77), (154.04 ± 30.52); (14.63 ± 3.57),(36.75 ± 3.67), (272.26 ± 66.11), (846.12 ± 243.36); (14.75 ± 5.65), (37.22 ± 11.27), (278.51 ± 37.42),(861.67 ± 369.29) ], Bax protein expression level [ ( 15.07 ± 0.83 ) %, ( 23.79 ± 3.99 ) %, (38.51 ± 1.58 ) %, (53.86 ±1.74)%;(15.43 ± 1.45)%,(36.11 ± 1.37)%, (56.86 ± 1.97)%, (76.09 ± 2.01)%; (15.20 ± 1.03)%,(35.25 ±1.09)%, (55.56 ± 2.17)%, (74.48 ± 2.85)% ] was respectively increased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05). The bel-2 mRNA expression level [ (443.00 ± 244.47), (156.79 ±53.18), (62.13 ± 13.66), (23.10 ± 6.44); (420.55 ± 110.77), (48.15 ± 10.02), (14.91 ± 6.53), (7.54 ± 2.62);(577.75 ± 123.22), (49.68 ± 10.11), (12.41 ± 1.28), (7.22 ± 1.89)], bcl-2 protein expression level[(47.20 ±3.77)%, (41.80 ± 2.94)%, (36.00 ± 2.36)%, (29.00 ± 2.91)%; (45.90 ± 4.15)%, (35.70 ± 2.77)%, (29.80 ±2.78)%, (24.80 ± 2.66)% ; (46.70 ± 3.47)%, (36.20 ± 2.90)%, (30.10 ± 3.21)%, (25.10 ± 2.28)% ] wasdecreased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05 ). In 3,9,15 mmol/L NaAsO2, apoptotic rate of p53 group, mRNA expression of bcl-2, protein expression of bcl-2 was higher than that ofnormal group and PC group, respectively (all P < 0.05), but mRNA expression of Bax, protein expression of Bax was respeetivelylower than that normal group and the PC group(P < 0.05 ). Conclusion p53 gene reduced the apoptosis induced by NaAsO2 in HELF, possibly by changing the apoptosis pathway.

AIMTo determine the entrapment efficiency of breviscapine nanoliposomes.

METHODSRetrodialysis method was used to determine the entrapment efficiency of breviscapine nanoliposomes. The drug recovery and the sample recovery were considered to verify the feasibility of the method.

RESULTSThe drug recovery was (99.6 +/- 0.6)%. The sample recovery was (99.9 +/- 0.8)%. The entrapment efficiency of breviscapine nanoliposomes was (75.3 +/- 2.3)%.

CONCLUSIONThe method used in this study is simple, applicable and accurate for determination of the entrapment efficiency of breviscapine nanoliposomes.

Asteraceae ; chemistry ; Drug Carriers ; Flavonoids ; administration & dosage ; chemistry ; isolation & purification ; Liposomes ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

The purpose of this study was to investigate the molecular mechanism by which miR-21 and its target genes mediate radiation resistance of glioblastoma cells. Real-time PCR was employed to detect miR-21 expression in normal brain tissues, glioblastoma tissues and glioblastoma cell lines (A172, T98G and U87MG). T98G cells were transfected with anti-miR-21 oligonucleotides, or plasmids containing PDCD4 or hMSH2 (PDCD4-pcDNA3 and hMSH2-pcDNA3). The survival curve was obtained to investigate the sensitivity of T98G cells to radiation. Cell apoptosis was measured by using the Caspase-3/7 kit and cell cycle by flow cytometry. Western blotting was performed to detect the expression of hMSH2 and PDCD4 in miR-21-inhibiting T98G cells. The results showed that miR-21 expression in glioblastoma cells and tissues was conversely associated with the radiation sensitivity. Over-expression of miR-21 resulted in radiation resistance, while knockdown of miR-21 led to higher sensitivity of glioblastma cells to radiation. After miR-21 knockdown, the apoptosis of T98G cells was significantly increased and the G(2) phase arrest was more significant. In addition, miR-21 knockdown increased the expression of endogenous PDCD4 and hMSH2, which contributed to the apoptosis and G(2) arrest of T98G cells. The findings suggested that miR-21 may mediate the resistance of glioblastoma cells against radiation via its target genes PDCD4 and hMSH2. MiR-21 and its target genes may be used as potential molecular targets for clinical radiotherapy sensitization in the future.
Apoptosis Regulatory Proteins ; genetics ; Cell Line, Tumor ; Glioblastoma ; genetics ; Humans ; MicroRNAs ; genetics ; MutS Homolog 2 Protein ; genetics ; RNA-Binding Proteins ; genetics ; Radiation Tolerance ; genetics

Objective To explore the effects of potassium iodate (KIO3) on the proliferation, cell cycle, and the mRNA/protein levels of cyclin D 1 and Ki67 of SW579 cells, a human thyroid squamous carcinoma cell line . Methods The effects of different doses of KIO3(0,10-6,10-5,10-4,10-3,10-2,10-1 mol/L)on SW579 cell prolifer-ation were assessed by CCK8 method.SW579 cells were then treated with 0 (control), 10-6 or 10-2 mol/L KIO3 for 48 h.The cell cycle was measured by flow cytometry .The mRNA and protein levels of cyclinD 1 and Ki67 were re-spectivelyanalyzedbyreal-timePCRandWesternblot.Results 10-6and10-2mol/LofKIO3respectivelyexhibi-ted the most promoting and suppressive effect on SW579 cell proliferation.G1 phase of cells in 10-6 group short-ened significantly( P<0.05) , and S phase prolonged significantly( P<0.05); while each phase of cells in 10-2 mol/L group changed in the opposite way( P<0.05) .KIO3 at the dose of 10-6 mol/L significantly up-regulated the mRNA levels of cyclin D1 and ki67 in cells( P<0.05) ;whereas, the mRNA and protein levels of cyclin D1 and Ki67 were significantly down-regulated in cells treated with 10-2 mol/L KIO3( P<0.05) .Conclusions Different doses of KIO3 affect the proliferation and cell cycle of SW579 cells probably by regulating the levels of cyclin D1and Ki67 .

To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

An introduction was first given to the advantages and application status of multi-rotor unmanned aerial vehicle (UAV) in the field of medical rescue. Then, the latest worldwide application and research status of multi-rotor UAV in this field were analyzed according to available research. On this basis, the application prospects of multi-rotor UAV in the scenarios of medical rescue field were assumed. Finally, it was pointed out that multi-rotor UAV would show more functions in medical rescue field owing to various technological development and breakthrough, signifying the bright deepening application prospect of multi-rotor UAV.

The analysis of poorly conductive samples using discharge plasma is a significant challenge in determining elemental content by atomic emission spectrometry.In this study,graphite as a conductive medium was combined with metal salts and silica to prepare soil sample pellets.A tungsten needle electrode was employed to generate point discharge microplasma with the sample,exciting atomic emission signals of analyte elements.By using Cu 324.7 nm as the analytical line,both the capability to excite atomic emission spectral lines and the impact of experimental conditions were investigated,resulting in accurate determination of trace copper in both standard and real samples.This method held promise for detecting other metals like manganese,iron,and zinc.This work offered methodology for non-conductive sample analysis via discharge plasma atomic emission spectrometry,showing potential for rapid on-site assessment of soil samples.

Objective To establish the differential expression profiles for exploring new immune mechanism of hepatitis B virus infection.Methods DCs were separated from the bone marrow of healthy mouse and cultured in vitro.Then DCs were stimulated with HBsAg,LPS,TNF-α,respectively.Twentyfour hours later,the total RNA of cell was extracted.cDNA microarray was used to compare the differential expression of RNA.The significant differential expression of miRNA after the stimulation of HBsAg was chosen.Subsequently,target genes of the significant differential miRNA were forecasted by using computer software.Results There were 30 miRNAs whose significant differential expressions were beyond two times after the stimulation of HBsAg.Among them,16 miRNAs expressions were increased and 14 miRNAs expressions were decreased.There was significant difference in differential expression of miRNA among the 3 different stimulations.The selected target genes included relevant elements of signal pathway of DC.Conclusion The alteration of expression profiles of miRNA was specific after DC was stimulated by HBsAg.The selected target genes further demonstrated that miRNA could play important roles in the immune mechanism of HBV infection.

Primary central nervous system germ cell tumors (CNS-GCTs) in children and adolescents have unique clinical features and methods of treatment compared with those in adults. There is little information about Chinese children and adolescents with CNS-GCTs. Therefore, in this study we retrospectively analyzed the clinical features and treatment outcome of Chinese children and adolescents with primary CNS-GCTs. Between January 2002 and December 2012, 57 untreated patients from a single institution were enrolled. They were diagnosed with CNS-GCTs after pathologic or clinical assessment. Of the 57 patients, 41 were males and 16 were females, with a median age of 12.8 years (range, 2.7 to 18.0 years) at diagnosis; 43 (75.4%) had non-germinomatous germ cell tumors (NGGCTs) and 14 (24.6%) had germinomas; 44 (77.2%) had localized disease and 13 (22.8%) had extensive lesions. Fifty-three patients completed the prescribed treatment, of which 18 underwent monotherapy of surgery, radiotherapy, or chemotherapy, and 35 underwent multimodality therapies that included radiotherapy combined with chemotherapy or surgery combined with chemotherapy and/or radiotherapy. PEB (cisplatin, etoposide, and bleomycin) protocol was the major chemotherapy regimen. The median follow-up time was 32.3 months (range, 1.2 to 139 months). Fourteen patients died of relapse or disease progression. The 3-year event-free survival (EFS) and overall survival rates for all patients were 72.2% and 73.8%, respectively. The 3-year EFS was 92.9% for germinomas and 64.8% for NGGCTs (P = 0.064). The 3-year EFS rates for patients with NGGCTs who underwent monotherapy and multimodality therapies were 50.6% and 73.5%, respectively (P = 0.042). Our results indicate that multimodality therapies including chemotherapy plus radiotherapy were better treatment option for children and adolescents with CNS-GCTs.
Adolescent ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; Central Nervous System Neoplasms ; therapy ; Child ; Child, Preschool ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; statistics & numerical data ; Disease-Free Survival ; Etoposide ; administration & dosage ; Female ; Humans ; Male ; Neoplasm Recurrence, Local ; Neoplasms, Germ Cell and Embryonal ; therapy ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo understand the characteristics of embB gene mutation of Mycobacterium tuberculosis (MTB) isolates from tuberculosis patients in Chongqing, and the value of embB306 as a molecular marker used to diagnose ethambutol (EMB)-resistant MTB strains.

METHODSDirect sequencing was used to analyze the polymorphism of embB mutation in 51 EMB-resistant MTB strains and 50 EMB-sensitive MTB strains. And diagnostic testing was used to evaluate the value of embB306 as a molecular marker of EMB -resistant MTB strains as compared with the traditional sensitivity test.

RESULTSAll 34 of 51 EMB-resistant strains (66.7%) and 3 of 51 EMB-sensitive strains (6%) had had embB306 mutation. The embB306 mutation rate in EMB-resistant strains coming from previously treated case was 87.5%, showing significantly higher than that from new cases (48.1%, P < 0.01); embB306 mutation rate was increased with the number of the resistant drugs; embB306 mutation serving as a marker to diagnose EMB-resistant MTB strains comparing with the traditional sensitivity test, had the rate of sensitivity = 66.7%, specificity = 94.0%, accuracy = 80.2% and Youden index = 60.7%.

CONCLUSIONembB306 mutation should be the main mechanism of MTB resistance to EMB in Chongqing, showing an association with the history of the treated and numbers of the resistant drugs. embB306 mutation should be a good marker to diagnose EMB-resistant MTB strains.

China ; DNA Mutational Analysis ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Pentosyltransferases ; genetics ; Tuberculosis, Multidrug-Resistant ; microbiology