Objective To evaluate the effect of subfascial endoscopic perforator surgery(SEPS) for the(treatment) of recurrent venous ulcer.Methods SEPS Was preformed on 56 limbs with recurrent venous(ulcer),and ulcer healing was assessed at follow up.Results Fifty-five limb ulcers healed in 10～49 days,one limb ulcer was not healed after 90 days,but healed after ligation of lesser saphenous vein.The patients were followed up for 1 to 3 years,1 case recurred after 6 months and it healed with antibiotic treatment;2 cases recurred after 18 months,and both healed after high ligation of lesser saphenous vein.Conclusions SEPS is effective and safe in the treatment of recurrent venous ulcer.SEPS is minitraumatic with few(complications).

BACKGROUND: Longterm therapeutic effects of routine drug treatment, intervention or vascular bypass transplantation on lower extremity arterial occlusion are not ideal. During recent years, angiogenesis of stem cells possibly becones a new method to repair or rebuild an effective collateral circulation at infarct regions.OBJECTIVE: To verify the effect of autologous bone marrow mesenchymal stem cell transplantation on promoting angiogenesis in rabbit ischemic hind limbs.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed in Jiangsu Institute of Schistosomiasis Control between August 2005 and November 2006. MATERIALS: Eight rabbits were used to prepare ischemic models of hind limbs, and then they were randomly divided into experimental group (n=4) and control group (n=4).METHODS: Bone marrow mesenchymal stem cells were isolated and cultured from New Zealand rabbits in the experimental group, and they were then marked 5-bromo-2-deoxyuridine (Brdu). Suspension of bone marrow mesenchymal stem cells was injected into the ischemic hind limbs in the experimental group, while the same volume of saline was injected into the controls. MAIN OUTCOME MEASURES: After two weeks, two-dimensional and color Doppler ultrasound detection was used on rabbit femoral artery to measure inner diameter of blood vessel, peak velocity and acceleration time of blood flow before and after transplantation. Muscle tissues were obtained from ischemic regions to observe distribution of transplant cells and state of angiogenesis using immunofluorescence staining and hematoxylin-eosin (HE) staining. RESULTS: Two weeks after transplantation, the inner diameter of femoral artery and the peak velocity of blood flow in the experimental group were higher than those in the control group (P < 0.01), but the acceleration time of blood flow was shorter than that in the control group (P < 0.01). Lmmunofluorescence staining showed that anti-Brdu-staining positive cells were found out in transplant part in the experimental group; while HE staining indicated that vessel density of ischemic region in the experimental group was higher than that in the control group. CONCLUSION: Bone marrow mesenchymal stem cells can promote angiogenesis, while autologous bone marrow mesenchymai stem cell transplantation will become a simple and effective method to treat lower limb ischemia.

Objective To explore the effects of H.pylori and crude extracted proteins secreted by H.pylori(broth culture filtrate protein,BCF-P)on acid secretion from isolated rabbit parietal cells.Methods Parietal cells from rabbit gastric mucosa were isolated and enriched with digestion and elutriation.H.pylori(NCTC 11637,CagA+ VacA+)were grown in liquid broth culture and BCF-P was precipitated with ammonium sulfate.The vacuolation activity of BCF-P was evaluated with neutral red dye uptake test in HeLa cell.Isolated parietal cells were incubated with H.pylori(bacteria/cell=100∶1)for 2 h and 16 h,or BCF-P(100μg/ml)for 1 h and 12 h.Acid secretion from parietal cells was studied using 14C-aminopyrine(14C-AP)accumulation indirectly and H+-K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results (1)BCF-P containing vacuolating cytotoxin(VacA)with vacuolation activity on HeLa cells had positive result on neutral red uptake test.(2)The basal expression of H+-K+ ATPase α subunit mRNA could be detected in isolated parietal cells and 14C-AP accumulation was significantly increased in response to the stimulation of histamine with different concentrations for 30 min(P＜0.05).These results indicated that the isolated parietal cells retain relative intact acid secretion function.(3)The histamine(1.0×104 mol/L)stimulated acid secretion was inhibited sustainedly in response to H.pylori by 81% at 2 h and by 94% at 16 h(P＜0.05).However,H+-K+ ATPase α subunit mRNA expression was up-regulated in tlle acute period(2 h)and was down-regulated in the chronic period (16 h)by H.pylori(P＜0.05).(4)BCF-P significantly inhibited the histamine-stimulated acid secretion by 24% at 1 h and by 58% at 12 h(P＜0.05),and this inhibition was accompanied by the down-regulated expression of H+-K+ATPase α subunit mRNA.Conclusions Intact H.pylori and VacA secreted by H.pylori could directly inhibit histamine-stimulated acid secretion from parietal cells and this inhibition may be mediated by the down-regulated H+-K+ ATPase expression.

Objective To analyze the mutations in keratin 1 (KRT1), KRT9 and KRT10 genes in a Chinese family with epidermolytic palmoplantar keratoderma (EPPK). Methods Clinical data were collected from a family with EPPK. Genomic DNA was extracted from the peripheral blood of 12 family members, including 6 patients and 6 unaffected members, as well as from 50 unrelated normal human controls. PCR was performed to amplify all the exons and flanking sequences of KRT1, KRT9 and KRT10 genes followed by DNA sequencing.Results A missense mutation C.1436T > C was found in the highly conserved helix termination motif of KRT1 gene of all the patients, resulting in a substitution of isoleucine by threonine at position 479 of the KRT1 protein. No mutation was found in the unaffected members or unrelated controls. Conclusions The missense mutation C.1436T > C in K.RT1 gene is likely to be the main cause of the phenotype of EPPK in this family.This is the first report of a pedigree with KRT1 gene mutation-induced EPPK in China.

Objective To study the disinfection efficacy of c ompound disinfectant of peracetic acid. Methods Suspended liqu id quantitative sterilization test and metal corrosion test were carried out wit h different concentrations of disinfectant compound. Results T he killing rate of Bacillus subtilis var.niger spores e xposed to peracetic acid 500 mg*L-1 or available chlorine 300 mg*L-1 for 15 minutes was 99.94% and 97.91% respectively, while that of the spores e xpos ed to the compound disinfectant containing both of them reached 100%. The influ ence of organic substances on the bactericidal efficacy of this compound disinfe ctant was less than that on the bactericidal efficacy of the single ingredient. The corrosive effect of the compound disinfectant on the metals was milder than that of peracetic acid, but heavier than that of dichlorodimethylhydantoin. Conclusions Disinfectant efficacy of compound disinfectant increase s clearly, while metal corrosiveness decreases.

Objective To investigate the potential mechanism of interleukin-1β(IL-1β)in H.pylori-related gastric carcinogenesis.Methods Human gastric cancer cell line(AGS),human gastric epithelial cell line(GES-1)and isolated parietal cells were treated with exogenous IL-1β(10 ng/ml)in the presence or absence of H.pylori(NCTC 11637).Cell proliferation and apoptosis were determined by MTT assay and flow cytometry,respectively. Expressions of cyclooxygenase-2(COX-2)mRNA and protein were examined using RT-PCR and flow cytometry,respectively.Acid secretion by parietal cells was tested using 14C-aminopyrine(14 C-AP) accumulation indirectly.H+/K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results The cell proliferation and H.pylori-related apoptosis in both GES-1 and AGS celL lines were stimulated and inhibited by IL-1β.IL-1β induced expression and upregulation of COX-2 mRNA in GES-1 and AGS cell lines,respectively.In addition,IL-1β continuously inhibited the ability of histamine-stimulated 14C-AP accumulation of isolated parietal cells accompanied by down-regulation of H+/K+ ATPase mRNA expression.Conclusions It suggests that IL-1β play an important role in H.pylori-related gastric carcinogenesis through two pathways:①to interfere gastric epithelial cell growth by up-regulating COX-2 expression,②to inhibit acid secretion from parietal cell by down-regulating H+/K+ ATPase expression.

BACKGROUND: There have been no effective means for clinical treatment of large regions of bone defects.Nano-hydroxyapatite/collagen(nHAC)composite would provide a new pathway for repair of bone defects owing to its similar structure to natural skeleton and better biocompatibility.OBJECTIVE: To investigate the role of nHAC composite co-cultured with bone marrow-derived mesenchymal stem cells(BMSCs)in repair of bone defects.METHODS: Following isolation and culture,human BMSCs were co-cultured with nHAC composite.Gross observation,histological analysis,and electron microscope observation were performed to analyze osteogenesis for repair of bone defects in the clinic.RESULTS AND CONCLUSION: Human nHAC could greatly proliferate in vitro.X-ray photography revealed that bone defects well healed after implantation of nHAC/BMSCs composite.These findings indicate that BMSCs exhibit osteogenic potential and nHAC is a satisfactory scaffold material for construction of tissue-engineered bone.

BACKGROUND: Bone bone bone marrow derived mesenchymal stem cells (MSCs) has the capacities of self-renewal and multi-directional differentiation, which can differentiate into osteoblasts after induction. Combined with basal hydroxyapatite materials, MSCs can repair the bone defect with satisfactory shape and function.OBJECTIVE: To observe the bone formation performance of segmental bone defect repaired by autologous transplantation after combination of MSCs and basal hydroxyapatite materials.DESIGN: Open experiment.SETTING: Department of Cells, Third People' s Hospital of Wuxi.MATERIALS: The experiment was conducted in the Department of Cells,Third People's Hospital of Wuxi between May 2004 and March 2005.Eighteen New Zealand rabbits of clean grade, aged 6 months were selected and randomly divided into cytoskeleton group, simple scaffold group and blank control group with 6 rabbits in each group. Nano-collagen basal hydroxyapatite bone were provided by the Department of Material, Tsinghua University, which is characteristic of natural-bone microstructure. Composition: about 57% were hydroxyapatite,and 30% were collagen and 13%were polylactic acid.METHODS: ①bone bone marrow derived MSCs of rabbits were isolated and cultured. Well-grown cells of the second generation were inspected of the expression of cell surface antigen (CSA) with EPICS-ALTRA flow cytometry. ②Scaffold materials were cut into blocks with the size of 6 mm ×6 mm×4 mm, sterilized by 60Co irradiance and infiltrated with DMEM-LG before using. Cells of the 2nd and 3rd generations were collected, cultured,the concentration of which was regulated to 109 L-1. Cells were co-cultured with nano-collagen basal hydroxyapatite in vitro. ③Model of segmental defects in radial of rabbits were established in all groups. The prepared compounds in the cytoskeleton compound group were embedded in the bone defect according to autograft principle, and blank scaffold materials were embedded in the bone defects of simple scaffold group. Nothing was transplanted in the blank control group. Rabbits in all groups received no internal or external fixation, whose soft tissues and skins were closely sutured.④The surface structure of materials and cell adherence in all groups were observed under scanning electron microscope (SEM).⑤Bone formation was studied by general observation, histological analysis and X-rays at 4, 8 and 16 weeks after operation respectively.MAIN OUTCOME MEASURES: ①The isolated culture and identification of MSCs.②Observation of all groups under SEM.③Results of radiological inspection of all groups. ④General morphous of cells in each group after operation.⑤Results of histological inspection of all groups after operation.RESULTS: A total of 18 enrolled New Zealand rabbits were involved in the analysis of results.①There were adherent cells at 4 hours after primary culture, which were like cloned at 48 hours after static culture. Cells were in irregular scale-shape with large cell body and the nucleus was in middle. Inspection with flow cytometer showed that CD29,CD44, CD-105 were positive, and the ratios of three positive cells were 97.4% ,98.1% and 86.2% respectively. ②The surface of simple scaffold group was irregular with many ventages. There were cells adherent to the surface of materials as well as the inside of ventage in the cytoskeleton compound group. ③The absorbance of X-rays at 4, 8 and 16 weeks after operation were higher in the cytoskeleton compound group than blank control group and simple scaffold group, moreover, bone union could be seen at the 16th week in the cytoskeleton compound group. ④The implants was embedded in the osteotylus at 16 weeks after operation in the cytoskeleton compound group,while parts of the middle segment of implants in the simple scaffold group was not covered by bone tissues. No porosis was found in the blank control group.⑤The ossifying capacity at each time point after operation was better in the cytoskeleton compound group than simple scaffold group, moreover,there was bone trabecula formed at the 16th week, and the osteoblasts were arranged in order. The bone tissues in the simple scaffold group mainly concentrated on the surface of materials, which sucked little. There were some bony tissues formed in the proximal end of defects in the blank control group, while none was seen in the middle parts.CONCLUSION: Rabbit MSCs can greatly proliferate in vitro, which has strong osteogenesis by being co-cultured with nano-collagen basal hydroxyapatite. It is a good kind of seed cells in tissue engineering.

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Objective To investigate the relationships of peroxisome proliferators activated receptor ?(PPAR?) and matrix metallo protease-7 ( MMP-7) with gastric cancer ( including low and high grade atypical hyperplasia). Methods One hundred and twenty gastric tissue specimens (four groups) were collected by gastroscopy or surgical operation, including 31 patients with chronic gastritis, 25 patients with low-grade atypical hyperplasia, 27 patients with high-grade atypical hyperplasia, and 28 patients with gastric cancer. The expression of PPAR? and MMP-7 were observed by immunohistochemical technique and analyzed by statistic methods. Results Both of them have high expression in the groups of gastric cancer respectively, and the high combined expressions of them were found in gastric cancer and high-grade atypical hyperplasia. Conclusion PPAR? and MMP-7 were the important differential signs of the gastric cancer and precancer by inference, the combined expression of PPAR? and MMP-7 was the evidences in differential diagnosis among gastric cancer, high and low grade atypical hyperplasia and gastritis.