Objective To investigate the correlation between the polymorphism of XRCC2 gene,a DNA double-strand break repair(DDSBR)gene,and the susceptibility to lung cancer in population of North China.Methods By PCR-based restriction fragment length polymorphism(RLFP)technique(PCR-RLFP),a case-control study was performed among 300 patients with lung carcinoma and 300 healthy controls to detect XRCC2(C41657T)polymorphism.Results There was no significant difference in XRCC2(C41657T)allele frequencies between lung cancer patients and healthy controls(P=0.16).The C/C,C/T and T/T genotype frequencies were 72.7%,24.0% and 3.3% in lung cancer patients,and 76.3%,22.0% and 1.7% in healthy controls,respectively.There was no significant difference in C/C and C/T genotype frequencies between cancer patients and healthy controls(?2=0.48,P=0.48).However,the C/T genotype frequency in non-smoke group was significantly higher in lung cancer patients than in healthy controls(?2=6.67,P=0.01).The risk for lung cancer in non-smokers was 2.11 times higher in C/T genotype carriers than that in C/C genotype carriers.There was no significant difference in C/C and C/T genotype frequency between lung cancer patients and healthy controls in the smoke group(P=0.16).Conclusion Overall,there was no significant correlation between genetic polymorphism of XRCC2(C41657T)and the susceptibility to lung cancer.But the C/T genotype might increase the risk of suffering from lung cancer in no-smoking populations of Northern China.

Adult ; Female ; Granuloma, Respiratory Tract ; pathology ; Humans ; Lung, Hyperlucent ; pathology ; Male ; Middle Aged

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Histone-Lysine N-Methyltransferase ; genetics ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Transcription Factors ; genetics ; Young Adult

Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.

OBJECTIVE: To determine the sensitivity of cetuximab induced apoptosis in laryngeal squamous carcinoma cells Hep-2, and to evaluate the synergistic killing effects and regulation mechanism of cetuximab alone or cetuximab in combination with chemotherapeutic agents (cisplatin) or radiation means on Hep-2 cells. METHOD: To investigate the cytotoxicities of cetuximab, cisplatin and radiation, cell counting kit-8 (CCK-8) assay was used for the detection of cell growth inhibition ratio, and fluorescence activated cell sorter FACS for the apoptotic rate and cell cycle distribution. RESULT: Cetuximab had inhibitive effect on Hep-2 cells within a certain range of concentration in a time- and dose-dependence manner. The inhibition concentration 50% (IC50) of cetuximab on Hep-2 cells for 24 h was 1 036.84 microg/ml. For application of cisplatin and radiation, the apoptotic rate of Hep-2 cell was higher by combining with cetuximab than their single or combined administration. Moreover, the cell cycle arrested at G0/G1 phase. CONCLUSION Laryngeal cancer Hep-2 cells was sensitive to the cetuximab induced apoptosis. Cetuximab combined with cisplatin and/or radiation can increase the antiproliferative effects on Hep-2 cells. These findings suggest the synergistic combination of cetuximab and cytotoxic agents was sequence depended.
Antibodies, Monoclonal, Humanized ; pharmacology ; Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Cetuximab ; Cisplatin ; pharmacology ; Combined Modality Therapy ; Humans

RNA interference (RNAi) has been proved as a novel approach for gene therapy. However, RNAi mono-therapy only aims at single gene, it therefore may ultimately fail to cure cancers caused by polygene variation. To overcome the deficiency of RNAi mono-therapy, "combinatorial RNA interference" (coRNAi) was put forward as a new strategy. By co-expressing the inducers of RNAi triggering single or multiple targets directly and other RNA- or protein-based silencers, coRNAi keeps target genes silent, prevents carcinogenic progression and induces apoptosis of tumor cells. This paper mainly reviews the major strategies of coRNAi and their applications in cancer gene therapy.
Animals ; Apoptosis ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; genetics ; Neoplasms ; genetics ; pathology ; therapy ; Oncogenes ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA, Small Nuclear ; genetics

Objective To establish a method of inducing dendritic cells(DC)from rat bone marrow cells in vitro,and identify the phenotype and function characteristics.Methods The rat bone malToW cells were collected and cultured in vitro under the condition of recombinant rat GM-CSF(rrGM-CSF)and recombinant rat IL-4(rrIL-4).After 2 weeks,the morphological character of DCs was observed under light microscope and scanning electron microscope.Expression of MHC-Ⅱ,CD80 and CD86 were detected by flow cytometry.The ability to stimulate allogenic T cells of the cultured DCs was detected by mixed lymphocyte reaction.Results DCs showed typical morphology with elongated dendritic processes under inversion microscope and scanning electron microscope.DCs at day 6 revealed immature phenotype,including MHC-Ⅱ(29.03 ±4.39)%,CD80(21.98±7.08)%and CD86(25.94±6.80)%.DCs at day 12 showed higher expression of MHC-Ⅱ(74.05±5.97)%,CD80(79.85±6.53)%and CD86(81.00±7.47)%,and stimulatory capacity of allogenic T cells,compared with that in DCs at day 6.Conclusion Matured DCs could be generated from rat bone marrow cells and attendance with rrGM-CSF and rrIL-4,which present the feasibility for further research on its application to allograft immunorejection.

Objective To study the roles of icariin and cyclic tensile strain (CTS) in promoting the osteogenic differentiation of adipose-derived stem cells (ASCs) and the molecular mechanisms involved.Methods ASCs were isolated from Sprague-Dawley rats and treated either with icariin (10-7 mol/L) or with 1000 μ,2000 μ or 3000 μ of CTS for 7 days,or with icariin plus CTS at 2000 μ for three days.Alkaline phosphatase (ALP) activity was detected after 3 and 7 days of intervention.Western blotting was performed to detect the expression of Runt-related transcriptional factor 2 (Runx2),Yes-associated protein (YAP) and connective tissue growth factor (CTGF) after the third day of the intervention.A reverse transcription polymerase chain reaction was performed at 7 days to detect the expression of osteopontin (OPN) and collagen la and after 3 days to detect the expression of the YAP target gene,CTGF and ankyrin repeating domain 1 (Ankrd1).Results Icariin and CTS at 1000 μ,2000 μ or 3000 μ could all significantly promote the expression of ALP protein.CTS at 2000 μ was the nost effective.The co-treatment with icariin and CTS significantly promoted ALP protein expression compared with icariin or CTS treatment alone.It also significantly promoted the expression of Runx2 and CTGF protein.Icariin or CTS (2000 μ) alone could not promote the expression of YAP protein,but icariin combined with CTS (2000 μ) promoted it significantly.Either icariin or CTS (2000 μ) could significantly promote ALP activity after 3 and 7 days,but icariin combined with CTS had the most obvious effect.Both icariin and CTS (2000 μ) could also significantly promote the expression of the osteogenesis-related genes OPN and collagen la,as well as the YAP targeted genes CTGF and Ankrdl.However,the combination of icariin and CTS had the greatest effect in promoting the expression of OPN mRNA,collagen la mRNA,CTGF mRNA and ankrdl mRNA.Conclusion Icariin and CTS co-treatment may promote osteogenic differentiation of ASCs via activating YAP expression.

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

Objective To investigate the mechanism of recurrent laryngeal nerve injury during total thyroidectomy.Methods The clinical data of 36 patients suffering from recurrent laryngeal nerve injury in total thyroidectomy from 2003 to 2014 was analyzed retrospectively.Results 21 cases had temporary recurrent laryngeal nerve injury while 16 cases had permanent injury.The cases of injury included giant thyroid goitre,with variation of recurrent laryngeal nerve,undergoing secondary surgery,with tumor invasion and complicating thyroiditis.During the first surgery,the incidence of temporary recurrent laryngeal nerve injury was 0.43％,and the incidence of permanent damage was 0.27％.The temporary and permanent injury incidence of secondary surgery after initial subtotal thyroidectomy was 4.59％ and 4.59％,respectively.For patients with grade 3 thyroid gland,that was 1.81％ and 0.30％,respectively and 0.51％,0.72％ respectively in malignant cases.In cases complicating thyroiditis temporary recurrent laryngeal nerve injury occurred in 1.01％.Conclusions There is increased risk of recurrent laryngeal nerve injury in giant goiter cases undergoing secondary operations.Hence it is suggested that initial surgical procedure be hemithyroidectomy or total thyroidectomy