AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.

AIM: To determine the effect of suction duration on thickness and diameter of corneal flap created by microkeratome in porcine eyes in laser in situ kerato-mileusis (LASIK).METHODS: Sixty porcine eyes were randomly assigned to three groups according to different suction durations: group 1 (10 seconds), group 2 (20 seconds), and group 3 (30 seconds). A Moria M2 microkeratome (Moria, France) with a 160μm head was used to create a corneal flap. Corneal flap thickness was measured by automated ultrasonic pachymetry, and the flap diameter was measured by a vernier caliper.RESULTS: The flap thickness of group 1, group 2 and group 3 was 146.05±13.46μm, 157.35±18.95μm and 169.25±21.02μm, respectively. There was a statistically significant difference among three groups (P=0.001). The mean flap diameter in groups 1, 2 and 3 was 8.63±0.19mm, 8.89±0.24mm and 9.06±0.18mm, respectively. A statisti-cally significant difference was found among groups (P<0.01).CONCLUSION: In LASIK in porcine eyes, an increase in suction duration resulted in a thicker and greater flap.

Background and purpose: Postoperative chemotherapy targets the metastatic cancer in the remaining lymph nodes, but the heterogeneity in multidrug resistance (MDR) of metastatic cancer cells is a main factor affecting chemotherapeutic efficacy. Recent studies only examined the primary lesion of esophageal squamous cell carcinoma(ESCC). There is no report about heterogeneity between the primary tumor and metastases lymph node. The purpose of this study was to explore the heterogenous expression and clinical signiifcance of multidrug resistance (MDR) associated proteins in primary tumors and metastatic lymph nodes in patients with thoracic ESCC. Methods:The expressions of lung cancer associated resistance protein (LRP), P-glycoprotein (P-gp), topoisomeraseⅡ(TOPO-Ⅱ), thymidylate synthase (TS), glutathione S-transferase-π (GST-π) were examined by immunohistochemistry in primary lesions and corresponding metastatic lymph nodes in 54 patients with thoracic ESCC. The differences between expression of primary lesions and matched metastatic lymph nodes were compared and analyzed in relationship with tissue differentiation degree. Results: The discordant rates of the expression and drug resistance between primary lesions and corresponding metastatic lymph nodes in LRP, P-gp, TS, TOPO-Ⅱ and GST-π were 63.0% and 26.9%, 42.6%and 22.2%, 48.1%and 25.9%, 50.0%and 29.6%, 18.5%and 1.9%respectively. The expression of LRP showed signiifcant difference between the primary tumors and lymph nodes (P=0.026). No signiifcant differences were found for the other four proteins, and GST-πwas expressed in all patients in both the primary tumors and lymph nodes. Protein expression was not associated with degree of differentiation. Conclusion:There is evident of heterogenous expression of MDR associated proteins in metastatic lymph nodes compared to the primary tumors of ESCC. The examination of expression levels of MDR associated proteins in metastatic lymph nodes is helpful to select the postoperative rational chemotherapy plan.

A series of novel sorafenib analogues were designed and synthesized. The cytotoxic activities of these compounds were tested in four tumor cell lines. Some of the compounds showed potent antiproliferative activity against the tested cell lines with IC50 = 4-20 micromol x L(-1). Some compounds demonstrated competitive antiproliferative activities to sorafenib against tested cancer cell lines. Among them, compound 7c demonstrated significant inhibitory activities on ACHN, HCT116 and MDA-MB-231 cell lines with IC50 values of 9.01, 4.97, 6.61 micromol x L(-1), respectively.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Inhibitory Concentration 50 ; Molecular Structure ; Niacinamide ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Phenylurea Compounds ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship

Objective To investigate the expression of FHIT gene in the 60Co gamma-ray irradiated human lymphocytoblast(AHH-1) cell and the bystander effect cell,and to explore the function of FHIT gene in the bystander effect of ionizing radiation.Method Preparation of bystander effect cell model:after irradiated with different dose of 60Co gamma-ray(0,2,5 Gy),the directly irradiated AHH-1 ceils were collected immediately by centfifugation and co-cultivated with non-irradiated cells in Transwell.forming the bystander effect group P1.In addition,some culture media supernatant of direcfly irradiated cells were transfefred to the non- irradiated cells culture medium,forming the group P2.Then cells were collected at 0,6,12,and 24 h after irradiation and the total RNA and protein were extracted.RT-PcR and Western blot were performed to determine the FHIT mRNA and protein level.respectively.Flow cytometry assay and cell counting were conducted to detect the alteration of cell cycle and cell proliferation,respectively at 0,24 h after irradiation.Results The mRNA level of FHIT gene among control cells,directly irradiated cells and bystander cells showed no obvious difference. while the FHIT protein level of the directly irradiated ceils and bystander cells was siguificandy down-regulated compared with the control cells(F=102.45,P<0.001).Moreover,the directly irradiated cells and bystander cells showed significant G2 phase arrest and obviously inhibited the proliferation ability.Conclusions 2 and 5 Gy of 60Co γ-ray irradiated AHH-1 cells can result in down regulation of the FHIT protein expression,which suggests that FHIT gene is involved in the process of bvstander effect induced by irradiation.

Adult ; Atropine ; therapeutic use ; Bronchodilator Agents ; therapeutic use ; Combined Modality Therapy ; Female ; Hemofiltration ; Humans ; Insecticides ; poisoning ; Male ; Middle Aged ; Organophosphorus Compounds ; Poisoning ; therapy ; Treatment Outcome

Animals ; Cochlea ; pathology ; Gene Deletion ; Hearing Loss ; etiology ; pathology ; KCNQ1 Potassium Channel ; genetics ; Mice

Objective To investigate the expression of macrophage migration inhibitory factor(MIF) in renal tissue of children with Henoch-Schonlein purpura nephritis(HSPN),and its correlation with clinical indexes and pathological changes,and to explore its effect on the pathogenesis of HSPN.Methods According to the clinical manifestation,60 children with HPSN were divided into only purpura group,mixed group and HSPN group.MIF concentration of Henoch-Schonlein purpura(HSP) groups and healthy control group were detected with enzyme linked immunosorbent assay(ELISA).MIF protein expression and the marker of human macrophage(CD68) in renal tissues of HSPN and normal control group were detected with immunohistochemistry method.The total urine protein for 24 hours and urinary N-acetyl-beta-D-glucosaminidase (NAG) level were detected with laboratory routine method.Results MIF concentration in mixed group and HSPN group were significantly higher than that in only purpura group and healthy control group(Pa

AIM: To investigate the effect of fetal bovine serum (FBS) on the proliferation and differentiation of murine corneal epithelial cells in vitro.METHODS: Mouse corneal epithelial cells(MCEs) were cultured in serum-free low-Ca2+ medium(KSFM) and KSFM supplemented with 100mL/L FBS, respectively. Population doublings (PDs) were determined. The expressions of corneal epithelial cell markers p63, keratin 19 (K19) and involucrin were investigated by RT-PCR and Western blotting analyses. RESULTS: Cells in KSFM were stably subcultured over 25 passages; however, none of the cell lines could pass P3 in KSFM with FBS. In KSFM, the cells showed typical cobblestone appearance and expressed p63, K19 and involucrin. After medium was supplemented with FBS, cells became homogeneous, large and squamous. Furthermore, both RT-PCR and Western blotting analyses showed that the expression of involucrin was increased significantly.CONCLUSION: FBS has effects of inhibiting proliferation and triggering differentiation of MCEs.

AIM:To investigate the roles of feeder cells ln stratification of murine corneal epithelial cells and build an ideal method to engineer stratified epithelial sheet.METHODS:Using contact feeder culture,separated feeder culture,compound feeder culture and culture without feeder cells by Air-lifting method in Transwell chamber culture system,tissue engineered corneal epithelium was reconstructed.Corneal sheets were stained with hematoxylin and eosin(HE)for histological observation.The expression of p63 and keratin 19(K19)and involucrin(IVL)was investigated by immunocytochemistry analysis.RESULTS:Stratification was limited to three to four layers in the contact feeder group,whereas separate feeder sheets were slightly more stratified.The compound feeder group produced a stratified epithelium with five to seven layers of cells.The group without 3T3 feeder cells formed only two to three layers of cells.Immunostaining images in the compound feeder group showed expression of progenitor markers p63 and K19 in the basal and suprabasal layer,as well as differentiation marker involucrin in all layers.CONCLUSION:The remarkable stratification as well as the Iimbal phenotype makes the compound feeder system a candidate tool for cultivating transplantable epithelial sheets.