Development Of Halosulphate Based Phosphors And It's Applications

Synthesis and characterization of halosulphate-based phosphors is important for thermoluminescence dosimetry (TLD), radiophotoluminescence dosimetry (RPL) and scintillator materials. The enhancement of luminescence output in halosulphate-based phosphors and it may be useful for lamp, solid-state lamp and radiation. Dosimetry by activator as well as sensitizer are well known properties. The combustion technique is not applicable for the synthesis of TLD phosphors due to very fine particles, which show less TL intensity, while sol-gel, solid-state diffusion, melt method and precipitation methods are applicable for TLD phosphors. Two halosulphates namely Na21( SO4 ) 7 F6 Cl and 2K3Ca2(SO4)3F were prepared and doped with Dy and Tm for different concentration .Halosulphate , Na21( SO4 ) 7 F6 Cl was prepared by wet chemical method and Halosulphate , 2K3Ca2(SO4)3F was prepared by solid state diffusion method . The characterization was done by X - ray diffraction ( XRD ) , Thermo luminescence (TL) was also studied . For Dy doped Na21( SO4) 7 F6 Cl , The peak was observed at 1200 C and shoulder at 1750C for 0.2 % molar concentration of Dy. and for 2K3Ca2(SO4)3F doped with Tm the shoulder peak was observed at 240 0 C and at 150 0C for 0.7 % molar concentration of Tm.

In vitro culture of human corneal epithelial stem cells using modified explant culture / 中国组织工程研究

BACKGROUND: Corneal epithelial stem cells, also known as limbal stem cells, are distributed in the basal layer of limbal epithelium. It is extremely difficult to deal with limbal stem cell deficiency or dysfunction that is caused by severe thermal burn, chemical burn, and chronic inflammation of ocular surface. At present, in vitro culture of corneal epithelial stem cells using tissue engineering technology followed by clinical transplantation is a new and effective therapeutic direction. OBJECTIVE: To explore the feasibility of serum-free culture of human corneal epithelial stem cells in vitro using modified explant culture method. METHODS: The remaining donor corneal tissues after keratoplasty (less than 8 mm in diameter) were obtained from Henan Eye Bank, and the outer and middle limbus were dissected under surgical microscope. Two culture methods were used to culture human corneal epithelial stem cells. In the conventional explant culture group, the limbal tissues were adhered to the dish with the epithelium being upward, then Keratinocyte-serum free medium (K-SFM) was added into dishes, followed by incubation at 37 °C in a 5% CO2 incubator. In the modified explant culture group, limbal tissues were dissected to immerse in the K-SFM culture medium and incubated at 37 °C in the 5% CO2 incubator for 12 hours. The limbal tissues were then adhered to the dish with the epithelium being downward. The day whenever the cells from the limbal tissues adhered to the dish was marked as the 1st day of culture, and changes in cell morphology and growth were recorded by phase contrast microscopy every day. Immunofluorescent staining was used to detect the expression of K3 and p63 in primary cells on the 5th, 10th and 14th day of the modified explant culture. RESULTS AND CONCLUSION: The mean early stage of growth in the modified explant culture group was shorter than that of the conventional explant culture group (P < 0.05), and the mean growth rate of the modified explant culture group was higher than that of the conventional explant culture group (P < 0.05). In the modified explant culture group, cells had a good growth state, and many cells with small size gathered together on the 10th day of culture. On the 14th day, cell clones were formed, and the cells in the clone showed uniform morphology. On the 5th day, K3 highly expressed, while p63 lowly expressed in primary cells. On the 10th day, both of K3 and p63 had an increased expression. On the 14th day, there was no significant increase in the K3 expression, but the expression of p63 increased significantly. In the in vitro serum-free culture condition, the modified explant culture could significantly promote the growth of corneal epithelial stem cells, and expand corneal epithelial stem cells in vitro, which could provide sufficient seed cells for enriching corneal epithelial stem cells and constructing human limbal multilayered epithelial sheets.

Comparación de parámetros hematológicos y morfología celular en muestras de sangre con EDTA K2 y EDTA K3 / Comparison of hematological parameters and cellular morphology in blood samples with EDTA K2 and EDTA K3

El EDTA es el anticoagulante de elección en los laboratorios de hematología para la conservación de la muestra de sangre total. Existen dos tipos, EDTA K2 y EDTA K3, y su diferencia radica en la cantidad de moléculas de potasio. Algunas guías sugieren que hay diferencias entre el anticoagulante EDTA K2 y el K3 para el proceso del hemograma; sin embargo, con las nuevas presentaciones de los tubos que traen las casas comerciales, no se tiene claro si en realidad aún hay diferencia entre los dos anticoagulantes, y si esto puede alterar el resultado del hemograma, tanto en el resultado cuantitativo, como en el cualitativo. Objetivo. Comparar los recuentos leucocitarios, la hemoglobina, el hematocrito, el volumen corpuscular medio, las plaquetas y la morfología celular en muestras de sangre periférica con EDTA K2 y EDTA K3, en diferentes tiempos (0, 1 y 2 horas). Materiales y métodos. Se realizó un estudio cuasi-experimental, multivariado, multifactorial, que tiene como unidad de análisis la sangre anticoagulada con EDTA K2 y EDTA K3, extraída de 53 individuos a través de un muestreo no probabilístico por conveniencia. Resultados. Al comparar los resultados del estudio morfológico por medio del extendido de sangre periférica y los datos cuantitativos del hemograma, se encontró que no hay diferencias estadísticamente significativas usando EDTA K2 o K3. Conclusión. Se evidenció que el uso del EDTA K2 o EDTA K3 como anticoagulante de elección, procesando las muestras en un tiempo adecuado después de su recolección, no afecta los parámetros cuantitativos del hemograma automatizado ni los morfológicos.

EDTA is the anticoagulant of choice in hematology laboratories for the conservation of whole blood samples. There are two types, K2 EDTA and K3 EDTA, and their difference lies in the amount of potassium molecules. Some guidelines suggest that there are differences between K2 and K3 EDTA for the blood analysis process. However, with the new collection tubes offered by the commercial suppliers, it is not clear if in fact there is a difference between the two anticoagulants that would result in changes in blood parameters and cell morphology. Objective. To compare leukocyte counts, hemoglobin, hematocrit, mean corpuscular volume, platelets and cell morphology in peripheral blood samples collected with K2 EDTA and K3 EDTA, at different times (0, 1 and 2 hours). Materials and methods. A quasi-experimental, multivariate, multifactorial study was carried out, with anticoagulated blood as the unit of analysis, either with K2 EDTA or K3 EDTA, extracted from 53 subjects through a non-probabilistic sampling for convenience. Results. There was no statistically significant difference when comparing results of the peripheral blood smear and the quantitative hematological parameters using K2 or K3 EDTA. Conclusion. The use of either K2 EDTA or K3 EDTA as the anticoagulant of choice, when processing samples within a suitable time after their collection, proved equally satisfactory for both quantitative and morphological parameters

Long Intergenic Non-Protein Coding RNA 665 Regulates Viability, Apoptosis, and Autophagy via the MiR-186-5p/MAP4K3 Axis in Hepatocellular Carcinoma

PURPOSE: Long intergenic non-protein coding RNA 665 (LINC00665) plays a vital role in the development of cancer. Its function in hepatocellular carcinoma (HCC), however, remains largely unknown. MATERIALS AND METHODS: The expressions of LINC00665, miR-186-5p, and MAP4K3 were determined by qRT-PCR. Cell viability and apoptosis were evaluated by MTT and flow cytometry, respectively. Autophagic puncta formation was observed by fluorescence microscopy. Bioinformatics analysis, luciferase reporter assay, RNA immunoprecipitation, and RNA pulldown were performed to identify associations among LINC00665, miR-186-5p, and MAP4K3. Western blot was utilized to examine the expressions of MAP4K3, Beclin-1, and LC3. Tumor growth was evaluated in a xenograft model. RESULTS: Elevations in LINC00665 were observed in HCC tissues and cells. The overall survival of HCC patients with high levels of LINC00665 was shorter than those with low levels. In vitro, LINC00665 depletion inhibited viability and induced apoptosis and autophagy. miR-186-5p interacted with LINC00665 and was downregulated in HCC tissues and cells. Upregulation of miR-186-5p inhibited viability and induced apoptosis and autophagy, which were attenuated by upregulation of LINC00665. MAP4K3 was found to possess binding sites with miR-186-5p and was upregulated in HCC tissues and cells. MAP4K3 depletion inhibited viability and induced apoptosis and autophagy, which were attenuated by miR-186-5p inhibitor. In vivo, miR-186-5p expression was negatively correlated with LINC00665 or MAP4K3 in HCC tissues, while LINC00665 was positively correlated with MAP4K3. LINC00665 knockdown suppressed tumor growth. CONCLUSION: LINC00665 was involved in cell viability, apoptosis, and autophagy in HCC via miR-186-5p/MAP4K3 axis, which may provide a new approach for HCC treatment.

'In Vitro' Comparison of K3XF and TFA Cyclic Fatigue Resistance in Continuous Rotation and Adaptive Motion / Comparación In Vitro de resistencia a la fatiga cíclica de K3XF y TFA en rotación continua y movimiento adaptativo

Abstract: Among the most common problems within dental professionals, is the fracture of rotary instruments of nickel and titanium root canal files (NiTi), when performing a procedure on a patient. Therefore, the objective of this work is to compare the in vitro resistance to cyclic fatigue of K3XF and TFA rotatory instruments, in continuous rotation and adaptive motion. For this, a static and cyclic fracture test was used and a fracture pattern was searched. The numbers of cycles until the fracture and the length of the fragments were determined. Of all the groups evaluated, K3XF-A exhibited the best resistance to cyclic fatigue. In future research, it will be performed an autoclaving and immersion cycles in sodium hypochlorite to be able to determine the separation causes of the rotary instruments. As a recommendation, the operator could significantly influence the decrease in resistance to cyclic fatigue due to the misuse of rotary instruments.

Resumen: Entre los problemas más comunes entre los profesionistas dentales, es la fractura de los instrumentos rotatorios de limas de canales radiculares de níquel y titanio (NiTi), cuando se lleva a cabo el procedimiento al paciente. Es por eso, que el objetivo de este trabajo es el comparar la resistencia in vitro de la fractura cíclica entre los instrumentos rotatorios K3XF y TFA, en rotación continua y movimiento adaptativo. Para esto, se utilizó la prueba de fractura cíclica y estática y el patrón de fractura fue evaluado. El número de ciclos hasta la fractura y la longitud de los fragmentos fue determinado. De todos los grupos evaluados, K3XF-A presentó la mejor resistencia a la fractura cíclica. Como trabajo a futuro, se utilizará ciclos de inmersión en hipoclorito de sodio y autoclaveado, para ser capaces de determinar las causas de separación de los instrumentos rotarios. Como recomendación, el operador puede influenciar significativamente la disminución de la resistencia a la fractura cíclica utilizando de una manera errónea el dispositivo.

Expression of MAP3K3 mRNA and its prognosis in human ovarian carcinoma / 临床与实验病理学杂志

Purpose To investigate the expression of mitogen-activated protein kinase kinase kinase 3 (MAP3K3) mRNA in ovarian carcinoma patients and to explore the correlation among its expression, clinicopathological features and prognosis. Methods The expression of MAP3K3 mRNA in ovarian carcinoma and fallopian tube tissues were detected by qRT-PCR, and the correlation between MAP3K3 mRNA expression and clinicopathological features was also analyzed. Whether MAP3K3 mRNA expression could be used as an independent predictor of prognosis for patients with ovarian carcinoma was further determined. Results The expression of MAP3K3 mRNA in ovarian carcinoma was significantly higher than that in fallopian tube tissues (P＜0.05). High expression of MAP3K3 mRNA was significantly correlated with FIGO stage and Challenge model of ovarian carcinoma (P＜0.05). Kaplan-Meier survival analysis showed that the disease-free survival time and overall survival time of patients with high MAP3K3 mRNA expression were shorter than those with low expression (34 months vs 52.2 months, P＜0.05.38.6 months vs 52.5 months, P＜0.05). Univariate analysis showed that high expression of MAP3K3 mRNA was a risk factor for poor prognosis of ovarian carcinoma patients (HR=4.198, 95％C/: 1.711 ～10.302, P＜0.05). Conclusion MAP3K3 mRNA is highly expressed in ovarian carcinoma tissues. Its high expression is associated with FIGO stage, Challenge model and poor prognosis of ovarian carcinoma, which may involve in the malignant transformation of ovarian carcinoma.

Comparison of Improvacuter EDTA Tube with BD Vacutainer EDTA Tube for Routine Hematological Analysis: Clinical Significance of Differences, Stability Study, and Effects of K₂ and K₃ EDTA

BACKGROUND: The type of blood collection tubes is an important pre-analytical factor that may affect test results. We compared the test results of the Improvacuter EDTA tube (Improve Medical, China) with those of the currently used BD Vacutainer EDTA tube (Becton Dickinson, USA) and investigated the effects of K₂ and K₃ EDTA additives. METHODS: Peripheral blood samples from 100 outpatients were collected into the aforementioned tubes. The samples were evaluated for 17 hematological analytes, hemoglobin A1c, and erythrocyte sedimentation rate (ESR). The results were analysed using the paired t-test for comparison. Bland-Altman plots and Passing-Bablok regressions were used for analytes with statistically significant differences in the comparison. If the differences were not within total allowable error, they were defined as clinically significant. For stability testing, the initial results were compared against those from samples preserved for 72 hours. White blood cell count, red blood cell count, platelet count, and mean corpuscular volume from both tubes were compared to ascertain the differences between K₂ and K₃ EDTA additives. RESULTS: Hematocrit, mean corpuscular volume, mean corpuscular haemoglobin concentration, and ESR showed statistically significant differences (P<0.05) between two tubes. However, these differences were not considered clinically significant. Most of the analytes presented statistically significant differences in stability test; however, they were not clinically significant either. Additionally, the differences in the hematological parameters shown in the outcome were not clinically significant, depending on the type of the EDTA additives. CONCLUSIONS: The results indicate that Improvacuter EDTA tubes showed satisfactory performance. We conclude that the tubes are suitable for common clinical hematological use.

Quantitative evaluation of apical extrusion of intracanal bacteria using rotary ProTaper, K3XF, twisted and hand K‑file system: An ex vivo study.

Aims: The aim of this study was to evaluate the number of intracanal bacteria extruded apically during root canal preparation using rotary ProTaper, K3XF, twisted, and hand K‑file system. Subjects and Methods: Seventy extracted single‑rooted human mandibular premolar teeth were used. Access cavities were prepared and the teeth were mounted in glass vials. Root canals were then contaminated with a pure culture of Enterococcus faecalis (ATCC 29212) and incubated at 37°C for 24 h. The contaminated roots were divided into four experimental groups of 15 teeth each and one control group of 10 teeth. Group 1: ProTaper; Group 2: K3XF; Group 3: Twisted file; Group 4: Hand K‑file; Group 5: Control group. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony‑forming units (CFU)/ml. Statistical Analysis Used: The obtained data were analyzed using the Kruskal–Wallis one‑way analysis of variance and Mann–Whitney U‑tests. Results: There was a significant difference between the rotary and hand instrumentation system related to the apically extruded intracanal bacteria. Conclusions: Both the rotary and hand instrumentation systems extruded intracanal bacteria through the apical foramen. K3XF file system showed least bacterial extrusion amongst all instrumentation groups.

Research progress of anti-tumor effect of vitamin C combined with vitamin K3 / 肿瘤研究与临床

Vitamin C (VC) and vitamin K3 (VK3) are widely used in the fields of antioxidant,immunity improvement and hemostasis,etc.In the 1 940 s,the antitumor effect of these two vitamins was found.Research shows that the combination regimen of VC and VK3 with a ratio of 100:1 produces a maximum of synergistic anti-tumor effect.Such combination reduces the overall dose of drugs,thus avoiding the increase in their adverse effects.The combination of VC and VK3 effectively kills a variety of tumor cells,such as breast cancer,ovarian cancer,prostate cancer,bladder cancer and leukemia,etc.This combined therapy has been patent and is in phase Ⅰ and Ⅱ clinical trials.The clinical studies show that it prolongs the survival time of patients with terminal cancer,while its adverse reactions is mild.Therefore,VC in combination of VK3 is of broad application prospect.This paper reviews the synergistic anti-tumor effects of VC combined with VK3.

Influência da sequência técnica do sistema K3 Endo na segurança e rapidez do preparo de canais curvos simulados / Influence of the sequence technique of K3 Endodontic system on the safety and speed of the preparation of simulated curved canals

O objetivo do presente trabalho foi comparar duas sequências diferentes na rapidez e segurança no preparo de canais curvos simulados. Foram empregados 30 blocos de acrílico com canais simulados que foram divididos em Grupo 1 – La Axxess 20 e 35/.06 25/.06, 20/.06, 25/.04 e 20/.04 e Grupo 2 – K3 Universal (VTVT) – 25/.10, 25/.08, 35/.06, 30/.04, 25/.06 e 20/.04. No preparo de todos os blocos empregou-se a velocidade de 250 rpm e torque de 1.4N/cm. Foram confeccionados dois pontos externos de amálgama para mensuração do desvio e adotou-se 17 mm como comprimento de trabalho. No Grupo 1, dilatou- -se até o instrumento 25/.04, enquanto que no Grupo 2 até o 25/.06, e cronometrou-se o tempo de preparo de cada bloco. Na análise do desvio, radiografias com lima k 15 (antes) e K 25 (depois) foram realizadas e digitalizadas, medindo-se o ângulo antes e depois no programa Digora for Windows 1.51. Os dados foram comparados pelo teste T-Student e Mann-Whitney. Os resultados mostraram que em ambas as técnicas não ocorreram diferenças significantes entre o ângulo antes e depois e que não ocorreu diferença significante entre os desvios médios do Grupo 1 (0,130) e do Grupo 2 (1,330). No tempo de preparo, o Grupo 1 foi significantemente mais rápido que o Grupo 2. No Grupo 1, ocorreu a fratura de 1 instrumento, enquanto que no Grupo 2 ocorrerem duas fraturas. Concluiu-se que ambas as técnicas são seguras para o preparo de canais simulados curvos, sendo que no Grupo 1 o tempo de preparo é menor

The objective of this paper was to compare two different sequences of simulated curved canals preparation regarding speed and safety. Thirty acrylic blocks with simulated curved canals were divided into Group 1 – La Axxess 20 and 35/.06, 25/.06, 20/.06, 25/.04 and 20/.04 and Group 2 – K3 Universal (VTVT) – 25/.10, 25/.08, 35/.06, 30/.04, 25/.06 and 20/.04. At the preparation of the blocks it was applieda 250 rpm speed and torque of 1.4N/cm. Two external points of amalgamwere made to assist the measurement of the deviation and the stipulated work length was 17mm. The instrument dilatation reached 25/.04 at Group 1 and 25/.06 at Group 2, and the preparation time of each block was registered. For the deviation analysis two radiographs, an initial with K-file 15 and a final with K-file 25 were obtained and digitalized, measuring the initial and final deviation using the softwareDigora for Windows 1.51. Data were compared statistically by T-Student test and Mann-Whitney test. The results showed that,for both techniques,there were no significant differences between initial and final angle, neither between average deviation at Group 1 (0.130) and Group 2 (1.330). Regarding preparation time, Group 1 was significantly quicker than Group 2. The breakage of one instrument occurred at Group 1 and two breakages at occurred at Group 2. It was concluded that both the techniques are safe to prepare simulated curved canals and at Group 1 the preparation was faster

Effect of internal stress on cyclic fatigue failure in K3

OBJECTIVES: This study aimed to evaluate the relationship between the cyclic fatigue of a K3 file and internal stress intentionally induced until the activation of the auto-stop function of the torque-controlled motor. MATERIALS AND METHODS: K3 (Sybron Endo) .04 and .06 taper, size 25, 30, 35, 40 and 45 were used in this study. To give the internal stress, the K3 files were put into the .02 taper Endo-Training-Bloc (Dentsply Maillefer) until the activation of the auto-stop function of the torque-controlled motor. The rotation speed was 300 rpm and torque value was 1.0 N.cm. K3 were grouped by the number of induced internal stress and randomly distributed to 4 experimental groups (n = 10, Stress 0 [control], Stress 1, Stress 2 and Stress 3). For measuring the cyclic fatigue failure, the K3 files were worked against a sloped glass block and time for file separation was recorded. Data was statistically analyzed Statistical analyses were performed using two-way ANOVA and Duncan post-hoc test at p < 0.05 level. RESULTS: Except .04 taper size 30 in Stress 1 group, there were statistically significant differences in time for file separation between control and all experimental groups. K3 with .04 taper showed higher cyclic fatigue resistance than those of .06 taper. CONCLUSION: In the limitation of this study, the cyclic fatigue of the K3 file was influenced by the accumulated internal stress from use until the auto-stop function was activated by the torque-controlled motor. Therefore, clinicians should avoid the reuse of the K3 file that has undergone auto-stops.

Comparison of apical transportation and change of working length in K3, NRT AND PROFILE rotary instruments using transparent resin block / 대한치과보존학회지

OBJECTIVES: The purpose of this study is to compare the apical transportation and working length change in curved root canals created in resin blocks, using 3 geometrically different types of Ni-Ti files, K3, NRT, and Profile. MATERIALS AND METHODS: The curvature of 30 resin blocks was measured by Schneider technique and each groups of Ni-Ti files were allocated with 10 resin blocks at random. The canals were shaped with Ni-Ti files by Crown-down technique. It was analyzed by Double radiograph superimposition method (Backman CA 1992), and for the accuracy and consistency, specially designed jig, digital X-ray, and CAD/CAM software for measurement of apical transportation were used. The amount of apical transportation was measured at 0, 1, 3, 5 mm from 'apical foramen - 0.5 mm' area, and the alteration of the working length before and after canal shaping was also measured. For statistics, Kruskal-Wallis One Way Analysis was used. RESULTS: There was no significant difference between the groups in the amount of working length change and apical transportation at 0, 1, and 3 mm area (p = 0.027), however, the amount of apical transportation at 5 mm area showed significant difference between K3 and Profile system (p = 0.924). CONCLUSIONS: As a result of this study, the 3 geometrically different Ni-Ti files showed no significant difference in apical transportation and working length change and maintained the original root canal shape.

Effect of P1k3 on the transcriptional activity of p73 in H1299 cells / 中国癌症杂志

Background and purpose: Protein kinase P1k3 could increase the transcriptional activity of p53. However, the effect of P1k3 on the transcriptional activity of p73 is still unknown. Our study was to investigate the effect of P1k3 on the transcriptional activity of p73. Methods: Luciferase reporter assay, RT-PCR and colony formation assay were adopted to study the effect of P1k3 and kinase-deficient P1k3 (K52R) on the transcriptional activity of p73 in p53-deficient human lung carcinoma H1299 cells. Results: Luciferase reporter assay showed that p73 increased the luciferase activities induced by p21~(WAF1) and Bax promoters (P<0.05). After co-transfection with p73 and P1k3, the luciferase activities induced by p21~(WAF1) and Bax promotors were significantly decreased in a dose-dependent manner as compared with the group that transfected p73 only (P<0.05). However, kinase-deficient Plk3 (K52R) had no significant effect on the luciferase activities induced by p21~(WAF1) and Bax promoters (P>0.05). RT-PCR showed that p73 increased the mRNA expressions of p21~(WAF1) and BAX (P<0.05). P1k3 decreased the expressions of p21~(WAF1) and Bax induced by p73 (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the expressions of p21~(WAF1) and Bax induced by p73 (P>0.05). Colony formation assay revealed that p73 decreased the colony formation of H1299 cells (P<0.05). P1k3 decreased the inhibitory effect of p73 on the colony formation of H1299 cells (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the inhibitory effect of p73 on the colony formation of H1299 cells (P>0.05).Conclusion: P1k3 can inhibit the transcriptional activity of p73, where as kinase-deficient P1k3 has no effect on the transcriptional activity of p73.

Application of fluorescent-activated cell sorting (FACS) technique in the research of rabbit limbal stem cells / 眼科研究

Objective The fluorescence-activated cell sorting (FACS) technique is a method for the identification and isolation of different cell populations.At present,the special surface marker for limbal stem cells has been not found yet.This study aimed to investigate the application of FACS technique in the research of rabbit limbal stem cells.MethodsCorneal limbal tissue was obtained from New Zealand white rabbits and cultured using the explant culture method in SHEM.Side population cells (SP cells) and non-SP cells were sorted from cultured rabbit limbal epithelium cells by FACS at a excitation wavelength 350 nm,and acquistion length 450 nm (blue light) and 675 nm (red light).The SP cells and non-SP cells were identified by detecting the expression of ABCG2 and K3/K12.The colony-forming efficiency of SP cells and non-SP cells were evaluated by the observation of cellular vitality with trypan blue staining.The percentage of colony formation was calculated as the colony number in various group/200×100%.ResultsIn 48-72 hours after primary culture,limbal epithelial cells migrated from the cultured tissue mass to form the mambrane-like structure and achieved 70%-80% confluence.The cells showed round,polygon and flattened shape.The proportion of SP cells in cultured limbal epithelial cells was 0.22%±0.09% with a colony-forming efficiency of 5.52±0.45% in SP cells and 0.78%±0.73% in non-SP cells,with a statistically significant difference between the two populations (t＝2.17,P＜0.01).After verapamil,an inhibitor of the expression of the ABCG2 protein,was added into the medium,the proportion of SP cells in the cultured limbal epithelial cells declined to 0.04%±0.006%.The SP cells presented a positive immunoresponse for ABCG2 and absence of immunoresponse for K3/K12,but a contradictory staining result was found in non-SP cells.ConclusionFACS can be applied in the research of limbal stem cells.

Apoptusis of SMMC-7721 cells induced by vitamin K3 via up-regulating Fus expression on the surface of cells / 中华检验医学杂志

Objective To investigate vitamin K3 (VK3) effect on apoptosis of human liver cancer cells and its mechanism. Methods The SMMC-7721 cells were cultured in the experiment. The inhibitory effects of VK3 on SMMC-7721 cells were tested by CCK-8. Morphological evaluation of apoptosis was performed by Hcechst33342 staining. The distribution of cell cycle and apoptosis, and the expression of Fas were assayed by flow cytometry. Fas mRNA expression were detected by RT-PCR. And the concentration of soluble Fas(sFasL) in the culture supernatant were measured by EIJSA. Results The inhibitory rates ofVK3 (at concentrations of 2,5,10,20,25 umol/L for 48 h) on SMMC-7721 growth were 33.8% ,50.1%,63.9% ,78.5% and 84.7%, respectively. Compared with the control group, the number of cells in the G0/G1 phase increased, while that of S phase decreased. The apoptotic cell rates were 18.75%, 25.80%,38.80% ,29.92% and 26.18% ,respectively. The apoptosis cells were strongly stained by Hcechst33342.On exposure to VK3 at the concentration of (2,5,10 umol/L) after 48 h, the mean fluorescence intensity ofFas on cell surface and the expression of fas mRNA and the concentration of FasL in the culture supematantin SMMC-7721 were increased, but they all decreased at the high concentration of VK3. Conclusion VK3can inhibit the proliferation of SMMC-7721 cells and induce apoptosis via up-regulating expression of Fas and sFasL.

A study on transportation of apical foramen after overinstrumentation by ProFile(R), ProTaper(TM) and K3TM in simulated canals with different curvatures / 대한치과보존학회지

This study was done to evaluate transportation of the apical foramen after 0.5 mm overinstrumentation by ProFile, ProTaper and K3 in simulated resin root canal. Sixty simulated resin root canal with a curvature of J and S-shape were divided into two groups. Each group consisted of three subgroups with 10 blocks according to the instruments used: ProFile(R), ProTaper(TM), and K3TM. Simulated resin root canal was prepared by ProFile, ProTaper and K3 with 300 rpm by the crown-down preparation technique. Pre- and post-instrumentation apical foramen images were overlapped and recorded with Image-analyzing microscope 100X (Camcope, Sometech Inc, Korea). The amounts of difference in width and dimension on overlapped images were measured after reference points were determined by Image Analysis program (Image-Pro(R) Express, Media Cybernetic, USA). Data were analyzed using Kruskal-Wallis and Mann-Whitney U-test. The results suggest that ProFile showed significantly less canal transportation and maintained original apical foramen shape better than K3 and ProTaper.

A comparison of canal centering abilities of four root canal instrument systems using X-ray micro-computed tomography / 대한치과보존학회지

The purpose of this study was to compare the centering abilities of four root canal instrument systems and the amounts of dentin removed after root canal shaping using them. The mesial canals of twenty extracted mandibular first molars having 10 - 20degrees curvature were scanned using X-ray micro-computed tomography (XMCT)-scanner before root canals were instrumented. They were divided into four groups (n = 10 per group). In Group 1, root canals were instrumented by the step-back technique with stainless steel K-Flexofile after coronal flaring. The remainders were instrumented by the crown-down technique with Profile (Group 2), ProTaper (Group 3) or K3 system (Group 4). All canals were prepared up to size 25 at the end-point of preparation and scanned again. Scanned images were processed to reconstruct three-dimensional images using three-dimensional image software and the changes of total canal volume were measured. Pre- and post-operative cross-sectional images of 1, 3, 5, and 7 mm from the apical foramen were compared. For each level, centering ratio were calculated using Adobe Photoshop 6.0 and image software program. ProTaper and K3 systems have a tendency to remove more dentin than the other file systems. In all groups, the lowest value of centering ratio at 3 mm level was observed. And except at 3 mm level, ProTaper system made canals less centered than the other systems (p < 0.05).

Different doses of vitamin K3 effect the expression of OPN of rats kidney / 中华泌尿外科杂志

Objective To investigate the effect of different doses of Vitamin K3 on the expression of osteopontin in rats kidney. Methods After different dose of Vitamin K3 were injected to different groups of rats feeded with same stone-inducing agent, the level of expression of OPN in renal tissues was observed with immunohistological staining. Results OPN expression located at distal convoluted tubule when the rats feeded without stone-inducing agent, and the OPN expression stain was extended to proximal convoluted tubule when the stone-inducing agent was feeded. Vitamin K3 can decrease the extension of OPN expression induced by the agent and more doses of Vitamin K3, more effects. Conclusions Vitamin K3 can decrease the expression of OPN in the kidney tubule of rats feeded with stone-induced agent.

Effect of vitamin K3 decreasing the urine oxalate excretion in rats / 中华泌尿外科杂志

Objective To investigate the effect of vitamin K3 decreasing the urine oxalate excretion in rats. Methods A total of 80 male Sprague-Dawley rats (body weight, 200-250g) were randomly divided into 8 groups, ie, control group, only vitamin K3 group, stone forming group, stone forming plus 4.0、3.0、2.0、0.8、0.4mg/d Vit K3 group. Each group is 10 rats respectively. The change of urine oxalate was observed. Results Vitamin K3 can reduce the 24h urine oxalate excretion in stone-forming group rats, but there were no effects in control group rats(P

Effect of vitamin K3 on inducing apoptosis in cell line SMMC-7721 of human liver cancer and expression of survivin gene / 临床检验杂志

Objective To observe the effects of vitamin K3(VK3) on cell growth and apoptosis in the cell line SMMC-7721 of human liver cancer and the changes in expression of survivin gene.Methods The inhibitory effect of VK3 on SMMC-7721 cells was examined by CCK-8 (cell counting kit).Morphological evaluation of apoptosis was performed by Hoechst33342 staining.The amount of apoptotic cell was assayed by flowcytometry.The changes of survivin gene expression were detected by RT-PCR.Results The inhibitory rate of VK3 (at concentrations of 2,5,10,20,25 mmol/L for 48 h) on SMMC-7721 growth was 3.8%,50.1%,63.9%,78.5%,84.7% respectively.The cells in apoptosis were strongly stained by Hoechst33342 compared with the cells in control group.The apoptostic rate of cells was 33.28%,63.97%,77.69%,87.30% and 92.40%.Following exposure to VK3 at the concentration of 2,5,10,20 and 25 mmol/L for 48h,the survivin mRNA expression in SMMC-7721 was downregulated.Conclusion VK3 inhibits the proliferation of SMMC-7721 cells and induce apoptosis.The mechanism may related to survivn gene.