BACKGROUND: Thyroid cancer stem cells are essential to the recurrence and metastasis of thyroid carcinoma. Leukemia inhibitory factor receptor (LIFR) shows a downward trend in a variety of malignant tumors, and its overexpression can inhibit the recurrence and metastasis of malignant tumors. OBJECTIVE:To explore the effect of LIFR on the stemness maintenance and lung metastasis of thyroid cancer stem cells in vivo. METHODS: Primary thyroid cancer cells TCLM were isolated from the lung metastases of a metastatic thyroid cancer patient. Serum-free suspension culture was used to form tumor cell balls. Flow cytometry was used to screen CD133+phenotype of metastatic thyroid cancer stem cell subpopulation TCLM-S. The overexpressed recombinant lentiviral plasmid containing LIFR and its negative control containing the empty plasmid were infected into thyroid cancer stem cells TCLM-S at the ratio of virus/cell number=20, and screened with 2.0 mg/L puromycin to construct TCLM-SLIFRand TCLM-Scontrolstem cells which stably expressed LIFR and its control. Real-time quantitative PCR (qRT-PCR) was used to detect the expression of LIFR in TCLM-SLIFRand TCLM-Scontrolstem cells. Flow cytometry was used to detect the percentage of CD133+phenotype cell subsets, western blot assay was used to detect the expression of tumor stemness related factors SOX2, Oct4, Nanog and tumor invasion and metastasis related proteins E-cadherin, matrix metalloproteinase (MMP)-2, MMP-7 in TCLM-SLIFRand TCLM-Scontrol stem cells. TCLM-SLIFRand TCLM-Scontrolstem cells were respectively injected into BALB/c nude mice by tail vein, and the lung metastasis model of thyroid cancer stem cells was constructed. The effect of LIFR overexpression on lung metastasis was observed. RESULTS AND CONCLUSION: Compared with TCLM-Scontrolcells, the expression of LIFR in TCLM-SLIFRcells was significantly increased, the proportion of CD133+phenotype stem cell subsets was significantly decreased, the expression of SOX2, Oct4 and Nanog were significantly decreased, the expression of E-cadherin was significantly increased, and the expression of MMP-2 and MMP-7 was significantly decreased. Moreover, the number of lung metastasis in nude mice given TCLM-SLIFRcells was significantly decreased as compared with those given TCLM-Scontrol cells.To conclude,LIFR overexpression can decrease the stemness and ability of lung metastasis in vivo.

The volatile components of roots and stems of Zanthoxylum nitidum were investigated by supercritical fluid carbon dioxide extraction (SFE-CO2) and gas chromatography-mass spectrometry(GC-MS). Thirty-one and fifty-one compounds were identified in the supercritical extracts from roots and stems of Z. nitidum, respectively, and total twenty-seven compounds were the common constituents. Among them, the major constituents in root and stem supercritical extracts were spathulenol (18.49 and 26.18%), n-hexadecanoic acid (14.24% and 12.79%), ar-tumerone (6.95% and 8.88%), oleic acid (8.39% and 5.71%) and hexanoic acid (4.39% and 7.78%). The in-vitro MTT assay showed that the volatile components of roots and stems of Z. nitidum did not exhibited any cytotoxic activity against human cancer Huh-7 and normal IEC-6 cells. These results indicated the same nature of the volatile constituents in the root and stem of Z. nitidum. This investigation may provide further evidence for expansion of medicinal parts of Z. nitidum.
Animals ; Cell Line, Tumor ; Cell Survival ; drug effects ; Chromatography, Supercritical Fluid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Mice ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Zanthoxylum ; chemistry

Objective To evaluate the significance of CT assessment for extraarticular anatomy in treatment of displaced intraarticular calcaneal fractures.Methods(1)Measurement of normal calcaneum 40 pieces of adult calcaneum specimen were measured,items of measurement included height of culmination of posterior facet and tuberosity,width of posterior edge of sustentaculum and tuberosity.(2)CT measurement of calcaneum.Transverse(axial)and coronal CT scanning were obtained from 20 feet with displaced intraarticular calcaneal and 20 normal feet as control.Following items were measured in CT scanning:the height of culmination of posterior facet and tuberosity,the coronal talocalcaneal angle,in coronal scanning,the width of posterior edge of sustentaculum and tuberosity,the axial calcaneocuboid angle,in axial scanning.Results(1)Measurement of height of calcaneum height of culmination of posterior facet and tuberosity of calcaneal specimen were(43.07?2.85)mm and(44.69?3.67)mm respectively,and these two items from CT scanning of normal feet were(42.84?1.66)mm,(43.40? 3.01)nun,and from CT scanning of feet with calcaneal fractures were(34.76?3.24)mm,(40.41? 3.69)mm.There was a statistically significant different between these two items for normal calcaneal specimen and for CT scanning of feet with ealcaneal fractures(P

Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance.Therefore,the development of methods for mutation detection characterized with straightforward,highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed.Although some of the currently available methods have shown very encouraging results,their discrimination efficiency is still very low.Herein,we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination,which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1％ within 20 min.The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15-38.48.The method is sequence independent,which assures a wide range of application.The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.

To evaluate the visual performance of the patients with traumatic corneal astigmatism, after the treatment of topography guided off-flap epipolis laser in situ keratomileusi (off-flap Epi-LASlK).
●METHODS: This prospective clinical study was comprised of 21 eyes of 21 patients with irregular corneal astigmatism caused by trauma, they were treated by off-flap Epi - LASlK from July 2012 to December 2013. The data included uncorrected visual acuity ( UCVA), best spectacle - corrected visual acuity ( BSCVA ), contrast sensitivity 1, 6mo before and after surgery; the healing area percentage of corneal epithelia, the healing time of corneal epithelia and pain score at 3d after surgery.
●RESULTS: Postoperative 1mo both UCVA and BSCVA were improved significantly than that before surgery (t =15. 703, 4. 351, P< 0. 05); Compared with the 1mo after surgery, UCVA at 6mo after surgery raised significantly (t= 6. 867, P <0. 05). There was no statistical significance between 6 and 1mo after surgery about BSCVA (t= 1. 497, P = 0. 140 ). After surgery, mean spherical equivalent (SE) was reduced from -2. 43±3. 02D to -0. 23±0. 49D (P<0. 05), and the mean cylinder was reduced from -1. 86± 2. 23D to - 0. 46 ± 1. 03D (P< 0. 05). Postoperative 1mo,4 kinds of spatial frequency and contrast sensitivity had no significant difference compared with the preoperative (P>0. 05 ). Postoperative 6mo except the 3c/ d spatial frequency, the remaining 3 spatial frequency contrast sensitivity compared with those before operation were significantly improved ( P < 0. 05 ). The healing area percentage of corneal epithelia was 92. 46% ±8. 24% (80% -100%) at 3d after surgery; The healing time of corneal epithelia was 3. 50 ± 1. 56d; Pain scores at 3 and 7d after surgery was 1. 54±1. 32 and 0. 04±0. 64, respectively.
●CONCLUSlON: Topography-guided off-flap Epi-LASlK is safe and effective in treating the patients with traumatic corneal irregular astigmatism. The operation can improve both the contrast sensitivity and the visual performance.

BACKGROUNDKeratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.

METHODSA K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer. RPMI-1640 supplemented with 10% fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44(+)/CD24(-) as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog, N-cadherin, and E-cadherin) was analyzed with real-time PCR.

RESULTSMuch higher percentage of CSCs was achieved with K-SFM: 17.3% for MCF-7 cells, 17.4% for SKBR-3, and 20.0% for primary breast cancer culture. Less than 1% CSC was achieved using RPMI-1640 supplemented with 10% FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4, ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.

CONCLUSIONK-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; Cadherins ; genetics ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Culture Media, Serum-Free ; Female ; Homeodomain Proteins ; genetics ; Humans ; Keratinocytes ; cytology ; Nanog Homeobox Protein ; Neoplasm Proteins ; genetics ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; genetics ; Real-Time Polymerase Chain Reaction

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.
Animals ; Cations ; Cell Line, Tumor ; Fatty Acids, Monounsaturated ; Genetic Therapy ; Genetic Vectors ; Humans ; Liposomes ; Luciferases ; Lung Neoplasms ; Mice ; Mice, Nude ; Peptides ; Quaternary Ammonium Compounds ; RNA Interference ; RNA, Small Interfering ; Transfection

AIM:To evaluate the optic nerve and axon impairment of relapsing - remitting multiple sclerosis ( RRMS) and neuromyelitis optica spectrum disorders ( NMOSD ) via detecting the peripapillary retinal nerve fiber layer (pRNFL) and the ganglion cell complex( GCC) thickness by optic coherence tomography(OCT).
METHODS: Retrospective case control study. Two hundred three cases were collected from August 2014 to January 2016 in Beijing Tian Tan Hospital. They were divided into four groups, including the normal group (n=60), the RRMS group ( n = 60 ), the NMOSD anti -aquaporin- 4 autoantibody seropositive( NMOSD- AQP4 -Ab seropositive) group (n= 48), and the NMOSD-AQP4-Abseronegative group (n = 35). All people were detected for the average and four quadrants ( superior, inferior, nasal, temporal) of pRNFL thickness and the average and two quadrants (superior, inferior) of GCC thickness with OCT. One way analysis of variance or nonparametric tests was used to compare the differences of pRNFL and GCC thickness between groups.
RESULTS: Comparing with the normal group, the average and all quadrants of pRNFL and GCC thickness in the RRMS, the NMOSD - AQP4 - Ab seropositive and the NMOSD-AQP4-Ab seronegative group were thinner (P<0. 01). Among them, the pRNFL and GCC thickness in the NMOSD- AQP4 - Ab seropositive group was the thinnest. Differences between groups in the pRNFL thickness:compared with the RRMS group, all quadrants of pRNFL and GCC thickness in the NMOSD-AQP4-Ab seropositive group were significantly thinner(P<0. 01); compared with the NMOSD- AQP4- Ab seronegative group, the inferior, nasal and temporal pRNFL thickness in the NMOSD-AQP4-Ab seropositive group were significantly thinner(P<0. 05), while the superior quadrant did not show significant differences( P > 0. 05); compared with the RRMS group, the superior pRNFL thickness in the NMOSD - AQP4 - Ab seronegative group was significantly thinner ( P < 0. 05), while the inferior, nasal and temporal quadrants did not show significant differences ( P > 0. 05 ). Differences between groups in the GCC thickness: compared with both the RRMS and the NMOSD- AQP4- Ab seronegative group, all quadrants of GCC thickness in the NMOSD -AQP4-Ab seropositive group were significantly thinner (P<0. 05); compared with the RRMS group, the superior GCC thickness in the NMOSD - AQP4 - Ab seronegative group was significantly thinner(P<0. 01), while the inferior quadrant did not show significant difference(P>0.05).
CONCLUSION: The optic nerve and axon impairment in NMOSD - AQP4 - Ab seropositive group was the most severe and the impairment in RRMS group was the least severe. The impairment in NMOSD - AQP4 - Ab seronegative group was between the former two, and could be more similar to that of RMMS.

This paper reported that the activation of oncogenes in human fetal esopha geal epithelium treated by alternariol (AOH). It was found that NIH/3T3 cells were transformed via transfeetion of DNA extracted from human fetal esophageal epithelium which was cultured and treated by 10?g/ml AOH in a short term in vitro. The efficiency of primary loci was 0.17 focus per ?g of DNA. In the secondary transfection, the efficiency was 0.58 focus per ?g of DNA (P

Objective To observe the effects of five flavonoids include rutin(RU),dihydromyricetin(DMY),hesperetin(HP),daidzein(DA)and hydroxysaffor yellow A(HYSA)on myocardiocyte apoptosis induced by H2O2 and to explore their relationships with Keshan disease and the possible mechanism.Methods Primary cultured cardiocytes of neonatal rats were randomly divided into control group,model group,and flavonoids preincubation group.The cardiocyte apoptosis was examined by fluorescent staining,the rates of apoptosis were detected by flow cytometry,the expression of Bcl-2 family proteins associated with apoptosis were observed:by Western blot.Results Compared with model group[(24.33±6.51)%],RU[(13.95±3.80)%],DA[(11.82±3.50)%],HYSA[(12.33±3.78)%]could decreased the rate of apoptosis(P＜0.05).The five flavonoids could up-regulate Bcl-2 expression,down-regulate Bax expression,and increase the Bcl-2/Bax ratio[RU(0.989±0.094),DMY(0.931±0.280),HP(0.980±0.095),DA(1.049±0.092),HYSA(1.031±0.039),vs model(0.490±0.046),the difference had statistical significances(P＜0.05)],but the Bcl-xl did not significantly changed(P＞0.05).Conclusions RU,DMY,HP,DA and HYSA have antiapoptotic effects on cardiomyocyte via regulating Bcl-2 and Bax,which gives us a hint in prevention and treatment of Keshan disease.