Objective To observe the expression and location of galactose β-1,4-glycosidic bonds (Gal β-1,4-GlcNAc) in keloid tissue,and to investigate the role of glycoprotein galactosylation in the formation of keloid.Methods This study included 10 keloid tissue specimens,7 hyperplastic scar tissue specimens,and 6 normal skin specimens.Lectin blot analysis was performed to measure the glycosylation level of glycoproteins,saturated picric acid-Sirius red staining followed by polarization microscopy to observe the type,expression and distribution of collagens in these specimens.The type Ⅰ/type Ⅲ collagen ratio was calculated.Immunofluorescence-based histochemistry was carried out by using Ricinus communis agglutinin I to analyze the expression and location of Gal β-1,4-GlcNAc in these skin samples,and double immunofluorescent staining to observe the colocalization of Gal β-1,4-GlcNAc and type Ⅰ procollagen α1.Results Compared with the normal skin tissue,the keloid tissue showed a significant increase in the expression of Gal β-1,4-GlcNAc in glycoproteins with a relative molecular mass of 30 000 to 40 000.Polarization microscopy revealed that there was a considerable expression of type Ⅰ collagen fibers,which amounted to (71.53 ± 4.03)％ in all the collagen fibers.The type Ⅰ/type Ⅲ collagen ratio was significantly higher in keloid tissue specimens than in normal tissue specimens (2.56 ± 0.53 vs.0.91 ± 0.11,P ＜0.05).Fluorescence microscopy showed that Gal β-1,4-GlcNAc was uniformly distributed in the membrane and cytoplasm of fibroblasts in keloid tissue,and the expression intensity of Gal β-1,4-GlcNAc in keloid tissue was notably stronger than that in normal skin tissue.There was a colocalization between Gal β-1,4-GlcNAc and type Ⅰ procollagen α1 in keloid tissue.Conclusions The expression of Gal β-1,4-GlcNAc,which is mainly observed in fibroblasts,is upregulated in keloid tissue,suggesting that Gal β-1,4-GlcNAc may be involved in the modulation of factors responsible for excessive fibre formation during the repair process of keloid.

Objective To investigate the expressions of advanced glycation end products (AGEs) and their receptors in keloid. Methods Serum and skin tissue specimens were collected from 20 patients with keloid, 20 patients with hyperplastic scar and 20 normal human controls. Fluorospectrophotometer was used to measure the serum level of AGEs, and immunohistochemistry and Western blotting to detect the expressions of AGEs and AGER in skin tissue specimens. Results The serum level of AGEs was (0.713 ± 0.098) AU/ml and (0.699 ± 0.077) AU/ml respectively in patients with keloid and those with hypertrophic scar, significantly higher than that in normal controls (0.179 ± 0.056 AU/ml, F = 283.82, P < 0.01 ). A positive expression of AGEs and AGER was observed in tissue specimens of keloid and hyperplastic scar, but not in the control specimens. As Western blotting showed, the expressions of AGEs and AGER were higher in tissue specimens of keloid and hyperplastic scar than in the control specimens (F = 18.04, 42.80, both P < 0.05), while no significant difference between keloid and hyperplastic scar tissue specimens was observed (P> 0.05). Conclusion There is a high expression of AGEs and AGER in keloid, which may contribute to the development of keloid.

Objective:To investigate the distribution of precancerous conditions and lesions of high-risk population in the high-in-cidence area of esophageal cancer in Ci County, Hebei Province. Methods:Esophageal cancer was detected early in 40 to 69 year old patients in Ci Xian through endoscopic screening data and endoscopic screening using iodine staining and indicative biopsy. The pa-tients were classified according to gender, age group, statistical esophageal precancerous condition, and lesion detection rate. Results:The analysis included 11 423 cases by screening queue, and the esophageal biopsy rate was 66.90%. The detection rates of squamous epithelium with mild, moderate, and severe dysplasia were 11.84%, 2.66%, and 1.04%, respectively. DCIS detection rate was 0.40%in patients with squamous cell carcinoma. The detection rate of the patients had been infiltrated by the squamous cell carcinoma was 0.04%.The rate of the squamous cell carcinoma within the mucosa was 0.37%.The rate of the infiltration squamous cell carcinoma was 0.17%. The detection rate of the hyperplasia above average severe dysplasia and cancer was 2.01%. Conclusion: High incidence of esophageal precancerous lesions was found in the Ci County aged 40 to 69. A large number of asymptomatic patients with cancer were detected. Age and sex are closely related to detection rate.

Objective To investigate the expressions of advanced glycosylation end products (AGE) in skin of mice with diabetes mellitus (DM) for different durations,and to evaluate their influence on collagen fibers.Methods Forty healthy 8-week-old male C57BL/6J mice were divided into DM group (n =20) and control group (n =20) to receive multiple intraperitoneal injections of low dose streptozotocin (50 mg/kg) and citric acid buffer (0.1 mol/L),respectively,for 5 consecutive days.Ten mice were sacrificed in each group on week 4 and 12 respectively after the last intraperitoneal injection,and full-thickness skin tissue samples were harvested from the middorsal region of each mouse.Then,hematoxylin-eosin (HE) staining was performed to observe histological changes,and total collagen content was estimated according to hydroxyproline content measured by an alkalinehydrolysis method.The cross-linking degree of collagen was determined by Edman degradation method using pepsin,the mRNA expression level of collagen type Ⅰ and Ⅲ by real-time quantitative PCR,the content of AGE by fluorospectrophotometry and Western blotting,and the level of malondialdehyde (MDA) by using a thiobarbituric acid method.Statistical analysis was carried out by t test.Results As light microscopy showed,the skin became obviously thinner in the diabetic mice with a progressive decrease in the number of collagen fibers in comparison with the control mice.On week 4 and 12 after the last injection,the diabetic mice exhibited a significant reduction in the content of hydroxyproline ((684.5 ± 76.7) vs.(787.7 ± 87.7) rg/g,(558.1 ± 73.1) vs.(757.8 ± 75.3) mg/g,both P ＜ 0.01) and in the levels of cross-linked collagen as well as mRNA expressions of collagen Ⅰ and Ⅲ (P ＜ 0.01 or 0.05),but a significant increase in the content of AGE ((37.47 ± 10.65) vs.(26.39 ± 3.74) AUF/mg hydroxyproline,(47.70 ± 5.66) vs.(29.91 ± 6.50) AUF/mg hydroxyproline,both P ＜ 0.01) and MDA ((6.62 ± 0.47) vs.(4.82 ± 0.56) μmol/L,(8.63 ± 0.36) vs.(5.15 ± 0.46) μmol/L,both P＜ 0.01) in skin tissue,compared with the control mice.The level of non-cross-linked collagen in skin tissue was also lower in the diabetic mice than in the control mice on week 12 (P ＜ 0.05).Moreover,the contents of hydroxyproline and the expression levels of collagen I in skin were significantly lower (P ＜ 0.05),but the levels of AGE and MDA were significantly higher (P ＜ 0.01) in the diabetic mice on week 12 than in those on week 4.Conclusions The characteristics of collagen fibers in skin are altered in diabetic mice when compared with normal control mice,which may be associated with increased AGE content and oxidative injury in skin.

Peripheral blood mononuclear cells (PBMNC) isolated from patients with acute leukemia (AL) and from normal controls were cultured in medium containing 1000 units/ml of recombinant interleukin 2 (IL-2). Marked LAK activity was induced on the third culture day in the normal controls,, with the highest cytotoxicity appearing between day 3 and 5 whereas induction of LAK activity in the AL patients began on the 5th day of culture, with the peak level appearing at day 15, showing that the peak of LAK activity was significantly delayed in AL. LAK cells surface phenotyping tests showed that CD8+ and CD16+ positive cells began to increase significantly from day 5 and reached the highest level at week 3, whereas CD4+ subclass began to decrease on day 5 and dropped to the nadir at week 3. The proportion of CD8+ and CD16+ cells were positively cor related with LAK activity, but, that of the CD4+ cell was inversely related with the LAK activity.

Aim To investigate the myocardial hypertrophy,the expression of transforming growth factor ?1(TGF-?1),Smad3 and Smad7 proteins in spontaneous hypertensive rats(SHR)and the effects of benazepril and candesartan.Methods SHRs of 12 weeks old were given benazepril and candesartan for 12 weeks.The tail arterial pressure was measured every two weeks.At 12 th weekend,cardiac configuration,heart mass index,area of cadiocytes,concertrations of AngⅡin plasma and myocardium,expressions of TGF-?1、Smad3 and Smad7 proteins were measured respectively.Results The arterial pressure,wall thickness,heart mass index,area of cardiocytes and the expressions of TGF-?1,Smad3 proteins increased in SHRs but were attenuated after the treatment of benazepril or candesartan.After the combined treatment,the synergistic effect could be observed.The levels of cardiac tissue and plasma AngⅡwere reduced.The expressions of Smad 7 were up-regulated after the treatment of benazepril or candesartan,while they were stable after the combined use.Conclusion There is a synergistic effect of attenuating myocardial hypertrophy in SHRs by combined use of benazepril and candesartan.It may be related to the regulation of Ang Ⅱ,decreasing the expressions of TGF-?1 and Smad3.

Aim To investigate the effect of irbesartan and spironolactone on expressions of connective tissue growth factor(CTGF),P-p38MAPK proteins during vascular remodeling in renovascular hypertensive rats(RHR).Methods Renovascular hypertension was induced by two kidney one clip(2K1C)operation.8 weeks later,RHRs were given irbesartan or(and)spironolactone for 8 weeks.After 8 weekstreatment,the morphometric measurements were performed in the mesenteric arterioles.Concertration of carboxyterminal propeptide of typeⅠprocollagen(PⅠCP)and N-terminal propeptide of type Ⅲ procollagen(PⅢNP)in blood serum was measured by enzyme linked immunosorbent assay method,and the media collagen area was assessed by collagne-specific Picro-sirius red staining with computerized video processing.Expressions of collagen type I,CTGF,P-p38MAPK proteins were detected using immunohistochemistry respectively.Results The arterial systolic pressure was attenuated significantly after the treatment of irbesartan,and this effect could not be enhanced by spironolactone.The media thickness over lumen ratio,media cross-sectional area over luminal area ratio of mesenteric arterioles,concertration of PⅠCP and PⅢNP,media collagen area,and expression of collagen typeⅠwere significantly increased in RHRs but ameliorated by irbesartan and spironolactone.Meanwhile,the expressions of CTGF,P-p38MAPK proteins were up-regulated in RHRs but blunted significantly after the treatment of irbesartan and spironolactone.The combined treatment had the synergic effects.Conclusions There is a synergistic effect of attenuating extracellular matrix(ECM)producing and amelioration of vascular remodeling(VR)by combined use of irbesartan and spironolactone.It maybe related to the expressions of CTGF and P-p38MAPK proteins down regulated by these two drugs.

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.

BACKGROUND: Beta-tricalcium phosphate (β-TCP)/alpha-calcium sulfate hemihydrate (α-CSH) artificial composite bone has a porous morphology and good biocompatibility, and it is helpful to improve the fusion rate in a spinal fusion model, which however has not yet been confirmed.OBJECTIVE: To investigate the preparation methods, biocompatibility and application effect of β-TCP/α-CSH composite bone in the spinal fusion model.METHODS: (1) Calcium sulfate dihydrate under certain conditions and at a proper temperature can be dehydrated to prepare α-CSH. Healthy bovine cancellous bone was decellularized, degreased and sintered under the certain condition and at the certain temperature to prepare β-TCP particles. Then, the β-TCP particles were dissolved in anhydrous ethanol, suspended, dried, and then used to prepare the β-TCP/α-CSH composite bone. Osteoblasts from the rabbit periosteum were co-cultured with the composite bone, and then cell morphology, adhesion and proliferation were observed. (2) Twenty New Zealand white rabbits were selected to make bilaterally posterolateral spinal fusion models of the multiple thoracic vertebrae, in which β-TCP/α-CSH composite bone was implanted into the left side (experimental group) and autogenous bone implanted into the right side (control group). The spinal fusion rate was compared between the two groups.RESULTS AND CONCLUSION: (1) Under the phase contrast microscope, a relatively small amount of L929 cells adhered to the composite bone after 3 days of co-culture, while the number of adherent cells became relatively dense. Under the scanning electron microscope, there were many crystalline particles on the surface of the composite bone,indicating a higher number of adherent cells on the composite bone surface. (2) The spinal fusion rate was increased at 4 weeks after implantation of β-TCP/α-CSH composite bone, which was significantly higher than that after implantation of autogenous bone (P < 0.05). (3) At 4 weeks after autogenous bone implantation, the bone trabecular bone was scarce and clumped, and newborn bone tissues and the boneless autograft bone were dominant. After 8 weeks after spinal fusion, the newborn bone tissues around the autograft were further increased. At 4 weeks after β-TCP/α-CSH composite bone implantation, there was no degradation of debris, but existed a few new bone tissues; at 8 weeks after implantation,the composite bone was surrounded by newborn bone tissues, and thickened trabecular bone and degradation of the composite bone were found. To conclude, the prepared β-TCP/α-CSH composite bone can achieve a higher fusion rate in the spinal fusion model.

Aim To observe the effects of benazepril and irbesartan on myocardial collagen in chronic heart failure ( CHF ) rats and the possible mechanisms. Methods CHF model in rats was made by partially banding abdominal aorta to induce pressure overload cardiac hypertrophy. The rats were given benazepril or/and irbesartan for 8 weeks. Systolic blood pressure ( SBP) was monitored by rat tail artery. The ultrastruc-ture of myocardium was detected by transmission elec-tron microscope. The content of myocardial hydroxyproline and amount of cross-linked collagen were measured by hydrolysis method. The expression of type Ⅰ and Ⅲ collagen protein in myocardium was investigated by immunohistochemistry. The expression of p-38 MAPK and p-p38 MAPK proteins was detected by Western blot. Results Compared with the model of CHF, there was no significant difference in SBP af-ter treatment. The content of hydroxyproline in myocar-dium, degree of cross-linked collagen, as well as ex-pression of type I collagen, type Ⅲ collagen and p-p38 MAPK proteins were significantly decreased after treatment with benazepril or irbesartan ( P <0. 05 ) , and the combined treatment of them had better effects. Conclusion There is a synergistic effect of attenua-ting the quantity and quality of myocardial collagen in CHF rats by combined use of benazepril and irbesar-tan, which may be related to the down regulation of p-p38MAPK protein expression.