Objective To investigate sequential expression of bone sialoprotein(BSP), alkaline phosphatase(ALP) and cementum attachment protein(CAP) in cementoblast(CB) during mineralized culture in vitro, and study the morphological and biologic characters of the CB in this process. Methods CB was seeded on the glass coverslips, and cultured for 6h, 12h, 1d, 2d, 3d, 4d, 5d, 6d, 8d, 10d, 12d and 16d, respectively. The expression of BSP, ALP and CAP proteins was detected using immunocytochemical method. Results 6 hours after plated, cementoblast expressed all of the three proteins. In the second and third days after plated, the cells became confluent and formed multiple layers, BSP and ALP expression decreased, and CAP did not be expressed at all. From the forth day, the cells formed cell nodules with mineralizing function. The cells in the nodules strongly expressed BSP,ALP and CAP, the cells around the nodule weakly expressed BSP and ALP, and did not express ALP. During the following 10 to 16 days, the cell nodules became mineralized nodules. Conclusion These results elucidated the changes of ALP, BSP and CAP expression as well as cell morphology during the CB proliferation, differentiation and mineralization in vitro, and provided some valuable information for studying the formation of cementum and CB proliferation, differentiation and mineralization in vivo.

Objective To study the ultrastrueture of 3 types of periodontal cells, including periodontal ligament cells (PDLC), os-teoblast cells (OB) and eementoblast cells (CB). Methods After culturing 3 kinds of periodontal cells - PDLC, OB and CB, we observed them with transmission electron microscope. Results There was rich rough endoplasmic reticulum and lots of microfilaments in the cyto-plasm of CB and PDLC cells. There was rich rough endoplasmic in the cytoplasm of OB cells. Conclusion The main characteristic ultra-structure feature of the bovine CB and PDLC was rich rough endoplasmic reticulum and microfilaments in the cytoplasm. Compared with CB and PDLC, OB contained fewer microfilaments in the cytoplasm.

Objective:To study the role of 14-3-3εand Cdc25B in germinal vesicle (GV)-stage arrest of mouse oocytes,and to pay foundation for further study on the molecular mechanism of PKA/Cdc25B/14-3-3εpathway in GV-stage arrest of mouse oocytes.Methods:The eukaryotic expression vectors of pcDNA3.1-ZEO-HA-14-3-3ε, pcDNA3.1-MYC-Cdc25B-WT, pcDNA3.1-MYC-Cdc25B-S321A, and pcDNA3.1-MYC-Cdc25B-S321D were transcribed into mRNA invitro.The mouse GV-stage oocytes were collected after superovulation and divided into no injection group,TE buffer microinjection group,14-3-3εmRNA injection group,14-3-3εmRNAs + Cdc25B-WT mRNA injection group,and 14-3-3εmRNA + Cdc25B-S321A mRNA injection group,14-3-3εmRNA+Cdc25B-S321D mRNA injection group.The protein expression levels of HA-14-3-3εand MYC-Cdc25B and the phosphorylation status of Cdc2-pTyr15 were observed by Western blotting method.The morphological changes and germinal vesicle breakdown (GVDB)rates of mouse oocytes were observed under phase-contrast microscope. Results:None of the oocytes in no injection group, TE buffer microinjection group, 14-3-3εmRNA injection group,14-3-3εmRNA + Cdc25B-WT mRNAs injection group and 14-3-3εmRNA + Cdc25B-S321D mRNA were able to undergo GVBD until at least 20 h after injection (P>0.05 );the GVBD rates of oocytes in 14-3-3εmRNA+Cdc25B-S321A mRNA group at 1 h (5.00%±0.68%),2 h (62.00%±3.56%)and 3 h (100.00%± 0.00%)after injection were significantly higher than those in no injection group and TE buffer injection group (P<0.01);the oocytes in 14-3-3εmRNA+ Cdc25B-Ser321A mRNA group at 20 h (79.00%±2.80%)after injection progressed to MII (P<0.01).Conclusion:14-3-3εcan regulate the transition from GV to GVBD of mouse oocytes by means of phosphorylation and dephosphorylation of S321-Cdc25B.

Alveolar Epithelial Cells ; pathology ; Diagnosis, Differential ; Humans ; Hyperplasia ; diagnostic imaging ; etiology ; metabolism ; pathology ; Lung Diseases ; diagnostic imaging ; etiology ; metabolism ; pathology ; Mucin-1 ; metabolism ; Nuclear Proteins ; metabolism ; Radiography ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism ; Tuberous Sclerosis ; complications ; diagnostic imaging ; metabolism ; pathology

Objective: Epithelial dysplasia of the esophagus and gastric cardia is precancerous lesion, including mild, moderate and severe levels. In 2000 year, WHO recommended to replace dysplasia with intraepithelial neoplasia. Mild and moderate dysplasia were classified as low-grade intraepithelial neoplasia (LIN). Cardia adenocarcinoma was suggested to be called esophageal-gastric junction adenocarcinoma. The risk of cancer development and the rule of time evolution were detected in esophagus and esophageal-gastdc junction LIN in high incidence area of esophageal cancer in Northern China, in an effort to provide scientific data for the prevention of esophageal cancer. Methods: Between October 2001 and October 2002, two townships of Cixian were chosen to carry out endoscopic iodine staining screening cohort study. The total population aged 0-85 was 22,016, of which 6,596 aged 40-69 (3257 males and 3339 females). Except for thoese with contraindications and those who refused to join the study, 3,506 cases were finally recruited in the study, and the screening rate was 53.2%. According to WHO criteria of the pathological diagnosis, the esophageal squamous epithelium with mild and moderate dysplasia and esophageal-gastric junction with mild dysplasia were classified into LIN groups (including 616 cases). The control group contained a total of 2,478 cases without precancerous lesions and free of cancer in endoscopic screening. Results: From June to September in 2008, the cohort was followed up and 174 cases were lost, with a follow-up rate of 95.0%. Follow-up was 3,970.7 person- years in the LIN group and 16,120.0 person-years in the control group.Carcinomous conversion rates were 251.7 and 68.2/per 100,000 person- years respectively in the LIN group and the control group. The median time in the two groups was 38 and 47 months, respectively. Compared with that of the normal population, the relative risk (RR) of LIN was 3.69 (95% CI=1.57-8.69, P=0.001). Conclusion: Population with LIN are at high-risk for esophageal cancer and endoscopic examination every year is absolutely necessary.

Objective To assess the continent diversion results of sigma rectum pouch after radical cystectomy. Methods The reconstruction of bladder with sigmoid was modified for treatment of 18 cases of bladder tumor.The intestine was incised over a length of 20～24 cm with the junction of sigmoid colon and rectum as the midpoint so as to create a low pressure reservoir for urine and side-to-side anastomosis was performed on the posterior borders of the rectosigmoid wall.Submucosal tunnel modified technique was em- ployed in antireflux urethral implantation,Urination has been controlled by anal sphincter.Results About 80 minutes was spent to finish a new low pressure pouch after radical cystectomy.Among 18 patients with this op- eration,the controlled emiction were good after pull out the anal duct and"J"stent in 1 week to 2 months.Af- ter 2 months,the times of urination is stable,4～5 times in daytime and 1～3 times during nighttime.Two pa- tients had nocturnal enuresis and the symptom vanished after 2 months. One patient had adhesive ileus, two patients had hyperchloremia acidosis and kaliopenia,one patient had urethral stump cancer.There is no com- plication as anastomotic block,renal function lesion and severe upper urinary tract infection. Conclusion This operative method was easy,emiction control was well,and with higher quality of life for patients.It is al- so a better alternative diversion procedure that would be easily accepted.

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo describe the incidence and mortality rates of esophageal cancer from 1974-2002 in Cixian county of Hebei province. Basic information on comparative geographical, epidemiological, and clinical research was collected.

METHODSIn early 1970s, cancer registry system in Cixian was established, collecting information on all the esophageal cancer cases in Cixian. Data was checked manually, then computerized, coded and analyzed using the software--SPSS 11.5.

RESULTSFrom 1974 to 2002, there were 18 471 esophageal cancer cases in Cixian, with 11 068 males and 7403 females, respectively. The age standardized incidence rate (ASR) for males was 208.77 per 100,000, while 120.47 per 100,000 for females. The trend of incidence rate of esophageal cancer had decreased during the 29 years from 1974 to 2002 (trend chi(2) = 19.94, P < 0.001). From 25 years of age onward, the incidence rates of the lower age groups declined with the increase of age. As for geographic distribution, the incidence rate in mountainous areas and hilly areas showed a significant declining trend in mountainous areas, chi(2) = 195.00, P < 0.001; hilly areas, chi(2) = 46.08, P < 0.001. The esophageal cancer incidence in plain areas remained steady, but had a slight increase in recent years. From 1969 to 2002, there were 18,736 cases died of esophageal cancer with 11 598 males and 7138 females. The ASR for male was 127.17 per 100,000 and 101.57 per 100,000 for female. Compared with the year 1969, the mortality rate of esophageal cancer in 2002 had a 37.96% decline. The proportion of esophageal cancer among malignant tumors in different decades decreased significantly.

CONCLUSIONThe trend of the incidence rate of esophageal cancer had been decreasing for the last 29 years. The incidence rate in mountainous areas and hilly areas showed a declining trend while in the plain areas it remained steady but having slight increase in the recent years. The mortality rate of esophageal cancer had a significant decrease from 1969 to 2002.

Adult ; Age Factors ; China ; epidemiology ; Esophageal Neoplasms ; epidemiology ; mortality ; Female ; Humans ; Incidence ; Male ; Registries ; Software

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

Cytotoxic T cells (CTLs) play a key role in the control of Hepatitis B virus (HBV) infection and viral clearance. However, most of identified CTL epitopes are derived from HBV of genotypes A and D, and few have been defined in virus of genotypes B and C which are more prevalent in Asia. As HBV core protein (HBc) is the most conservative and immunogenic component, in this study we used an overlapping 9-mer peptide pool covering HBc to screen and identify specific CTL epitopes. An unconventional HLA-A2-restricted epitope HBc141-149 was discovered and structurally characterized by crystallization analysis. The immunogenicity and anti-HBV activity were further determined in HBV and HLA-A2 transgenic mice. Finally, we show that mutations in HBc141-149 epitope are associated with viral parameters and disease progression in HBV infected patients. Our data therefore provide insights into the structure characteristics of this unconventional epitope binding to MHC-I molecules, as well as epitope specific CTL activity that orchestrate T cell response and immune evasion in HBV infected patients.
Adult ; Amino Acid Sequence ; Animals ; Binding Sites ; Epitopes ; chemistry ; immunology ; metabolism ; Female ; Genotype ; HEK293 Cells ; HLA-A2 Antigen ; metabolism ; Hepatitis B Core Antigens ; chemistry ; immunology ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Hydrogen Bonding ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Middle Aged ; Molecular Dynamics Simulation ; Mutation ; Protein Binding ; Protein Structure, Tertiary ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism