2.Study of nanofiber microcarriesr/collagen gel on in vitro cultivation of hepatocytes
Wei HU ; Xin LIU ; Hanqing GU
International Journal of Biomedical Engineering 2012;(6):321-324
Objective To investigate cell's viability and functions when human hepatocyte cell HepG2 and microcarriers were embedded in collagen gel and cultured in vitro.Methods Ultrasound concussion crushing method was used to get nanofiber microcarrier.HepG2 and microcarriers were embedded in collagen gel and cultured in 12 d.The albumin (ALB),urea and lactate dehydrogenase (LDH) levels in the nutrient solution were measured.Results The cells in nanofiber nanofiber microcarrier/collagen gel group were gathered around the microcarrier and formatted aggregates,the ALB and urea levels increased steadily and reached maximum in day 9,then decreased; In collagen gel without nanofiber microcarrier group cells uniformly distributed,the ALB and urea levels reached maximum in day 3,then decreased rapidly,and on day 6 majority of cells dead.Conclusion Gel entrapped nanofiber microcarriers and hepatoeytes is a high-density and longtime culture method which can possibly be used in the bioartificial liver system.
3.Analysis of stage and clinical pathological characteristics of invasive lobular carcinoma with the 8th AJCC Cancer Staging system
Hui HU ; Yang ZHAO ; Wei WEI ; Xin YI ; Yinhua LIU
International Journal of Surgery 2017;44(5):328-331
Objective To study the meaning of breast cancer staging system by AJCC eighth edition to invasive lobular carcinoma and analysis the clinical pathological characteristics.Methods According to the eighth edition of the AJCC staging to evaluate the TNM stage and prognosis evaluation of invasive lobular carcinoma cancer patient in Peking University Shenzhen Hospital from 2011 to 2016,and compared with others in clinical pathological data.Results There were 21 cases of invasive lobular carcinoma,accounting for 2.7% of all invasive breast cancer.We found that invasive lobular carcinoma shows no significant difference (P > 0.05) in ages,menstrual status,molecular features and anatomic staging and prognosis staging with others;histological grade were significantly different (P < 0.05).There were significant differences in the prognosis and staging of invasive lobular carcinoma.Conclusions Eighth AJCC staging systemn provides a new reference for the clinical treatment of breast cancer,should be evaluated with anatomic stage.Histological grade is relatively good in invasive lobular carcinoma and the prognosis is good,needs more research to the individualized treatment of invasive lobular carcinoma.
4.The Technique of Metallic Self-expanding Stents in Treating Malignant Gastroduodenal Obstruction
Xin WEI ; Liming ZHONG ; Hong HU ; Jian YOU ; Dacheng REN ; Xin HU ; Rongqiang PAN
Journal of Practical Radiology 2010;26(2):255-257
Objective To explore the methods and curative effect of metallic self-expanding stent in inoperable malignant gas-troduodenal obstruction. Methods The data of 15 cases with gastroduodenal obstruction including 9 cases of carcinoma of head of pancreas and 6 cases of carcinoma of stomach were analyzed retrospectively. The operative procedures of the stent implanted and the tors accepted more radiation dose because the manipulation was under the fluoroscopy in a short distance and with a full field of view. sions, the postoperative eating habit and the development turnover of disease. The main death reasons were tumor transfer and sys-tem exhaustion. Conclusion To pay close attention to the details and main points of operative procedure is the key point to implant stent successfully for malignant gastroduodenal obstruction. The determinative factor to influence the curative effect is the develop-ment turnover of tumor.
5.Protective effect of all-trans retinoic acid on injury of human immortalized hepatocytes induced by sodium arsenite
Hui-xin, SUN ; Xin-xin, HU ; Wei, ZHANG ; Yan-hui, GAO ; Dian-jun, SUN
Chinese Journal of Endemiology 2012;31(3):263-266
ObjectiveTo investigate the protective effect of all-trans retinoic acid (ATRA) on injury of human immortalized hepatocytes (HHL-5 cells ) induced by sodium arsenite and possible mechanisms.Methods After cultured for 48 h,HHL-5 cells were divided into four groups:normal group,ATRA group,sodium arsenite group and ATRA + sodium arsenite group.HHL-5 cell viability was tested by using cell proliferation experiment (WST).Superoxide dismutase(SOD),glutathione peroxidase(GSH-Px) activity,malondialdehyde(MDA) content,and aspartate aminottransferase (AST) activity in each group were determined by biochemical method.The microstructure of HHL-5 cells in each group was observed under transmission electron microscopy.ResultsHHL-5 cell viability(0.57 ± 0.02) of sodium arsenite group was compared with that of normal group(0.70 ± 0.01 ),the difference was statistically significant(P < 0.05).Levels of SOD,GSH-Px,MDA and AST[ (153.84 ± 2.35),(0.08 ±0.02)U/mg Prot,(4.15 ± 0.50)nmol/mg Prot,(265.43 ± 4.62) × 103 U/L] of sodium arsenite group were compared with that of normal group[(237.41 ± 18.30),(0.93 ± 0.02)U/mg Prot,(2.26 ± 0.40)nmol/mg Prot,(177 ± 9.85) ×103 U/L],and the difference was statistically significant (all P < 0.05).HHL-5 cell viability (0.65 ± 0.04) of ATRA + sodium arsenite group was compared with that of sodium arsenite group, and the difference was statistically significant (P < 0.05).Levels of SOD,GSH-Px,MDA and AST[ (286.85 ± 3.39),(0.56 ± 0.09)U/mg Prot,(3.36 ± 0.37)nmol/mg Prot, (220.02 ± 1.07) × 103 U/L] of ATRA+ sodium arsenite group were compared with that of sodium arsenite group,the difference was statistically significant(all P < 0.05).Compared with normal group and ATRA group,the surface microvilli of HHL-5 cells of sodium arsenite group decreased,double-membrane structure was unclear,vacuolar degeneration was seen in the cytoplasm,and glycogen was aggregated.The damage level of ATRA + sodium arsenite group was decreased.ConclusionsATRA plays a protective role through increasing intracellular antioxidant enzyme activity of HHL-5 cells,removal or reduction of oxygen free radicals produced by sodium arsenite.
6.Effects of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2 genes in HaCaT cells
Xin-xin, HU ; Yan-hui, GAO ; Wei, ZHANG ; Hui-xin, SUN ; Dian-jun, SUN
Chinese Journal of Endemiology 2012;31(4):365-368
Objective To observe the influences of different doses of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2(Nrf2) genes in HaCaT cells.Methods Cell proliferation was evaluated by Cell Counting Kit-8(CCK-8) assay after the HaCaT cells were exposed to 0.00,3.13,6.25,12.50,25.00,50.00,75.00,100.00 μ mol/L sodium arsenite for 48 h,respectively.Based on the previous results of cell proliferation,0.00(control),6.25,12.50,and 25.00 μmol/L of sodium arsenite were selected to treat HaCaT cells for 48 h,respectively.The mRNA expression of keratin 1,keratin 10,involucrin,loricrin and Nrf2 were detected by real-time fluorescent quantitative PCR.ResultsCompared with the control group (100.05%),HaCaT cell proliferation rates(83.06%,51.04%,39.52%,24.51%,16.99% and 9.04%) were significantly lower in 6.25,12.50,25.00,50.00,75.00 and 100.00 μ mol/L of sodium arsenite groups and the 50% inhibiting concentration was 12.38 μmol/L.Compared with the control group( 1.06 ± 0.28,1.00 ± 0.12,1.00 ± 0.08),the mRNA expression of keratin 1,involucrin and loricrin (0.08 ± 0.04,0.13 ± 0.12,0.05 ± 0.03;0.47 ± 0.11,0.21 ± 0.09,0.10 ± 0.15; 0.50 ± 0.27,0.31 ± 0.10,0.57 ± 0.23) were significantly decreased(all P < 0.05) in HaCaT cells treated with 6.25,12.50,25.00 μmol/L sodium arsenite,respectively.But keratin 10 mRNA expression showed a rise trend and the 6.25 μmoL/L sodium arsenite group (1.83 ± 0.45) was significantly higher than that of the control( 1.07 ± 0.14,P < 0.05 ).The Nrf2 mRNA expressions of HaCaT cells in 12.50,25.00 μmol/L sodium arsenite groups(0.13 ± 0.07,0.69 ± 0.33) were significantly lower than that of the control ( 1.00 ± 0.09,all P < 0.05 ).ConclusionsThe cellular proliferation and keratinization are decreased when HaCaT cells are exposed to sodium arsenite,which may be regulated by lowering Nrf2 mRNA transcription.
7.Dynamic expression changes of Smad3 and type Ⅰ collagen in sclera of progressive myopia in guinea pigs
Bo, JIANG ; Zhangyou, WU ; Zicheng, ZHU ; Wei, HU ; Xin, ZHAN
Chinese Journal of Experimental Ophthalmology 2017;35(6):491-497
Background Sclera remodeling process in axial elongation is one of the main pathological mechanisms of axial myopia progression.Studies confirmed that transforming growth factor-β1(TGF-β1) participates in the sclera remodeling process,and Smad3 is one of TGF-β1 downstream signal gene transcriptive factors,so to explore its role in sclera remodeling process of myopic eyes is of great significance for pathogenesis and prevention research of myopia.Objective This study was to investigate the expressions of type Ⅰ collagen and Smad3,a TGF-31 downstream target,in sclera of form deprivation myopic (FDM) eyes and explore the impact of TGF-β1-Smad3-type Ⅰ collagen signaling pathway on collagen remodeling in myopic sclera.Methods Seventy-five 1-week-old guinea pigs were randomly divided into normal control group (25 guinea pigs) and FDM group (50 guinea pigs).Monocular FDM was induced by occluding the left eyes of guinea pigs in FDM group with translucent latex balloons for 2,4,6 weeks,respectively,and consecutive occluding for 4 weeks followed by uncovering for 1 week (4/-1 weeks).The refractive power was detected by retinoscopy and axial length was measured with A-type ultrasound.Immunohistochemistry and reverse transcription-PCR were employed to detect the dynamic expressions of type Ⅰ collagen and Smad3 protein ad mRNA in the sclera of guinea pigs with emmetropia and experimental myopia,ard the relationship between collagen Ⅰ and Smad3 levels was analyzed.Results The refraction was hypermetropic in both normal control group and FDM group before occluding of eyes (P>0.05),and the hypermetropic power was gradually reduced over time in the normal control group.In the FDM group,the refractive power was gradually changed from (+2.09 ± 0.31)D before occluding to (-1.23±0.69),(-4.17±0.59),(-7.07±0.56) and (-4.30±0.58)D,and the axial length was increased from (5.93-±0.39)mm to (6.62±0.36),(7.30±0.34),(7.99--0.32),and (7.21 ±0.36) mm at weeks 2,4,6,and 4/-1 after occluding,respectively,indicating significant differences in refractive power and axial length over time in the FDM group from normal control group and self-control group (all at P<0.05).The expressions of Smad3 and type Ⅰ collagen protein and mRNA in the sclera of the FDM group was significantly lower than those of the control group and self-control group in various time points (all at P<0.05).The positive correlation were found in the expression of Smad3 on the myopic sclera with that of type Ⅰ collagen in both protein and mRNA levels (protein:r=0.993,P<0.05;mRNA:r=0.954,P<0.05).Conclusions The myopic power and ocular axis increase dependent upon occluding time,and the expressions of Smad3 and type Ⅰ collagen in the sclera are correspondingly weakened in FDM eyes.A consistent expression trend is found between Smad3 and type Ⅰ collage,suggesting Smad3 and type Ⅰ collagen participate in the regulation of sclera remodeling in myopia by TGF-β1-Smad3-Collagen Ⅰ signaling pathway.
9.The role of cell density in the morphology of the Candida albicans bioflims development and the tyrosol production
Xin WEI ; Guanling WU ; Weihong LIU ; Yan ZHANG ; Qin HU
Chinese Journal of Microbiology and Immunology 2010;30(4):344-348
Objective To study the role of cell density in the tyrosol production and morphology for Candida albicans biofilms. Methods C. albicans SC5314 and clinical isolates were propagated in yeast peptone dextrose (YPD) medium. Cells were collected by centrifugation and washed twice in sterile phosphate-buffered saline (PBS) before this study, then resuspended in RPMI 1640 supplemented with L-glutamine and adjusted to a desired concentration of 5 × 10~6 cells/ml, 5×10~5 cells/ml, 5 × 10~4 cells/ml, 5 × 10~3 cells/ml after counting with a hematocytometer. Standardized C. albicans cells were prepared as above description and 2000 μl of this standardized cell suspension was dispensed into the wells, then C. albicans biofilms were formed on the bottom of the polystyrene wells. In this study, tyrosol synthesized by SC5314 and clinical isolates of C. albicans biofilm was quantified by high performance liquid chromatography (HPLC). The effects of tyrosol on morphology of C. albicans biofilms were investigated by scanning electron microscopy (SEM). Results Tyrosol production of C. albicans biofilms was affected by cell densities. At lower inoculation size(5 μ 10~3 cells/ml), there was too less tyrosol production to be detected at the early stage of the biofilms formation. At higher inoculation size (5 μ10~6 cells/ml), tyrosol can be detectable at the early stage or at the mature stage of biofilms formation. There was a sharp increase in tyrosol concentration at 24 h, while there was a decrease in tyrosol concentration after that time from the strains when the strains were at an inoculation size of 5 × 10~6 cells/ml and 5 × 10~5 cells/ml. Cell densities affected the morphology formation of the C. albicans biofilms. At the early stage of the biofilms formation, C. albicans grew less germ tube at lower cell densities than that at the higher cell densities. With the mature of the biofilms, C. albicarts grew more hyphae at higher cell densities than that at the lower cell densities. All these above showed that cell densities played an important role in the propagation for the C. albi-cans in the biofilm formation. Conclusion Cell density play an important role in the formation of the C. albi-cans biofilms and the production of the tyrosol.