ObjectiveThis study is designed to investigate the relationship between STR polymorphism in HO-1 gene promoter and susceptibility of CAD,in order to provide a new strategy for prevention and treatment of CAD by using HO-1.Methods200 patients who were diagnosed as CAD by coronary angiography were selected in this study.100 subjects without evidence of CAD under coronary angiography with their sex and age similar to CAD patients were selected as controls.Genotyping was performed using polymerase chain reaction followed by capillary electrophoresis automated DNA sequencer.Each size of the (GT) n repeat was calculated using the GeneMapper Analysis software.ResultsA (GT)n polymorphism was found in the HO-1 gene promoter with n =16 ～39.Subjects with n≤29 expressed much more HO-1 protein than those with n ＞29( P ＜0.01 ).The alleles were then classified into two subgroups,S'allele (n 29 ) and L'allele (n ＞29),the subjects were then classified as having an S/S,S/L,or L/L genotype.Subjects with the L allele ( L/L + L/S genotypes) had more chance to get CAD than those with S/S genotype ( adjusted OR =1.83,95 ％ CI =1.04 - 3.24).Stratified analysis further showed that L allele ( L/L + L/S genotypes) was susceptive to CAD in patients who smoke (adjusted OR =2.59,95％ CI =1.16 -5.80).ConclusionsThe (GT)n polymorphism in HO-1 gene promoter is related to susceptibility to CAD,especially in those patients who smoke.

Objective To investigate the relationship between hypoxia-inducible factor-1α(HIF-1α) and construction of collateral circulation in patients with coronary artery disease. Methods Collateral vessels were determined in 96 patients with≥70% narrowing of at least one coronary artery without prior revascularization,42 patients with coronary artery collaterals and 54 patients with no coronary artery collaterals.Another 50 cases with normal coronary arteries were selected as control.The levels of HIF-lα expression in monocytes and lymphocytes were tested by immunohistochemistry (IHC) and Western blot.Results Compared to the controls,the patients had higher expression of HIF-1α(P<0.01).Higher HIF-1α expression was found in patients with collaterals than in those without collaterals (P<0.01).Positive correlation was observed between the expression of HIF-1α protein and collateral score (r1=0.78,r2=0.84,P<0.01).Conclusion HIF-1α expression in circulating monocytes and lymphocytes are associated with collateral circulation.Detection HIF-1α might be helpful in the prognosis of patients with coronary artery disease.

Objective To examine the expression of p44/42 mitogen -activated protein kinase (p44/42MAPK) and Phospho-p44/42MAPK in cicatrix (hy pertropic scars and keloids) and to aim at exploring the role of activated p44/ 42 MAPK pathway in development of cicatrix. Methods In or der to analyse the differences, 10 samples of normal skin (NS), hypertropic scar s (HS) and keloids (K) were collected, and then the extracted cytoplasmic protei ns from each tissue were examined by Western blotting for p44/42 MAPK and Phosph o p44/42MAPK and immunohistochemical staining with specific antibodies was emplo yed to determine that in fibroblasts of K, HS and NS. Results There was no evident difference of p44/42MAPK in the tissues and fibroblas ts between cicatrix and NS, but Phospho-p44/42MAPK was obviously higher in cica trix than that in NS. In cicatrix, there was no evident difference of p44/42MAPK in tissues between HS and K, while in fibroblasts, Phospho-p44/42MAPK in K was much higher than in HS. Conclusion Activation of p44/42MA PK pathways involves in formation of cicatrix.

Objective To explore the mechanism of inhibitory effect of dimethyl sulfoxide (DMSO) on the hyperplasia of the fibrous capsule around the tissue expander. Methods The experimental model was established in rats as follows: after implanting the expander on the back of rats, 30 % DMSO or normal saline was injected into the expander. The former was classified as experimental group, and the latter as control group. The cystic wall was resected after the skin and soft tissues were expanded. In situ hybridization and the immunohistochemical staining were used to determine the expression of typesⅠand Ⅲ of collagen and procollagen mRNA in the cystic walls. Results It was found that both the typeⅠand Ⅲ of collagen content and the expressing quantity of mRNA of procollagen in the cystic wall of the experimental group were less than those of control group. Conclusion The results imply that the mechanism that DMSO inhibits the expression of typesⅠand Ⅲ of collagen in the fibrous cystic wall may be achieved through down-regulating the genetic expression of procollagen in the fibroblasts.

Objective To investigate the changes and relationships between the serum endogenously produced interleukin-10(IL-10),nitric oxide synthase(NOS) and inducible nitric oxide synthase(iNOS) in acute myocardial ischemia and reperfusion.Methods Forty-two rats were randomly divided into two groups: myocardial ischemia and reperfusion group(the control group) and methylprednisolone treatment group.The rats in the control group and methylprednisolone treatment group received intravenous injection of placebo and methylprednisolone before ischemia respectively.The plasma IL-10,NOS and iNOS were detected at 0.5 h after ischaemia and 0.5h,2 h after reperfusion,respectively.Result The plasma IL-10,NOS and iNOS gradually increased at 0.5 h after ischaemia and 0.5h,2 h after reperfusion in the two groups.The levels of plasma IL-10 were significantly higher in the methylprednisolone treatment group in different time points than those in the control(all P

Objective To investigate the expression of nestin in keloid and hypertrophic scarring.Methods Samples were obtained from patients with normal skin(n=8),flat scars(n=7),keloids(n=8) and hypertrophic scars(n=8),respectively.Immunohistochemistry staining was used to measure the nestin expression in above samples.Nestin positive cells were counted to evaluate the expression levels.Results ①Nestin expression was detected in basal cells,prickle cells,sweat glands,hair follicles,sebaceous glands,and vascular endothelial cells in 8 samples of normal skin,and in fibroblasts in 7 of them.Nestin was also positively expressed in abovementioned cells or skin appendages in samples of flat scars,keloids and hypertrophic scars.②Nestin positive cell counts(epidermic cells,fibroblasts,and vascular endothelial cells) in keloids and hypertrophic scars were significantly higher than those in normal skin and flat scars(P0.05).Conclusions The increased expression of nestin in epidermic cells,fibroblasts,and vascular endothelial cells in keloid and hypertrophic scarring supports a possible role of nestin as fibrosis inducing factors in keloids and hypertrophic scars.

Objective To investigate the expression of nestin in fibroblasts of adult human dermis.Methods A total of 6 samples of normal human skin were collected.Immunohistochemistry and immunocytochemistry staining were used to detect the nestin expression in adult human dermis and in cultured fibroblasts of different passages(3rd,5th,7th,10th,and 12th passages,respectively) in vitro.Nestin positive cells were counted to evaluate the expression quantity and intensity.Results There were 103.3?67.4 fibroblasts per square millimeter(mm~2) of adult human dermis,and the nestin positive fibroblasts accounted for(9.5?3.0)% of total amount.The amount of nestin positive fibroblasts in vitro was obviously higher than that in vivo.Significant difference was observed in the amount of nestin expression among different passages(P

0.05),there were more apoptosis-starting cells found in normal skin(1738.33?348.89/mm2)than in keloid(891.67?395.00/mm2)(t=-5.565,P=0.000).The number of positive stained fibroblast cells in dermis significantly increased in keloid group(911.67?323.61/mm2),compared with that in normal skin group(220.00?80.00/mm2)(t=7.188,P=0.000).This result that the protein expression level of Bid was higher in keloid(0.46?0.08)than that in normal skin tissue(0.02?0.01)(t=18.905,P=0.000)was proven by western blotting test.Conclusions The number of apoptosis-starting cells in epidermis of normal skin is more than that in keloid.Bid protein in dermis of keloid is over expressed.

Objective To investigate whether nestin-positive cells can be isolated from human fetal pancreases and cultured in vitro for passages. Methods The islet-like cell clusters (ICCs) were isolated from pancreases of induced human fetuses at 16, 18, 20 gestational weeks and cultured subsequently. Immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of the nestin, pancreatic and duodenal homeobox gene-1 (PDX-1/IPF-1), islet endocrine hormone insulin and glucagon. Results Nestin-positive cells, which may further transform into sphere cell clusters that include PDX-1, insulin and glucagon expressing cells, as well as nestin-positive cells, were isolated from the ICCs in vitro and cultured with several passages. Conclusions The nestin-positive cells can be isolated from fetal pancreases, and may have the capability to proliferate and differentiate into islet endocrine cells.

Objective To explore the effect of Xiao Ban Xu, a compound Chinese traditional medicinal prescription, on the expression of collagenⅠand Ⅲ of keloid fibroblasts in vitro. Methods Six samples of keloid fibroblasts (KFB) and 6 samples of normal fibroblasts (NFB) entered the study as experimental and control groups, respectively. With the application of Xiao Ban Xu (10?g/ml), the fibroblast culture system and the ABC immunocytochemical staining method were used to investigate the expression of collagenⅠand Ⅲ of KFB and NFB. Results The density of the staining of collagenⅠand Ⅲ in the experimental group was much higher than that in the control group, with (7675 4?825 5 vs 2305 2?320 4 and 10595 2?1311 5 vs 2434 8?356 9; t =13 37 and 12 66, P =0 00004 and 0 00005) or without (11113 1?1304 9 vs 3519 6?236 0 and 11157 7?1300 3 vs 2626 5?426 3; t =14 42 and 13 47, P =0 00003 and 0 00004) the addition of Xiao Ban Xu. Compared with blank control group, the density of the staining of collagenⅠand Ⅲ was degenerated greatly in the experimental group after 48 hours of application of Xiao Ban Xu (7675 4?825 5 vs 11113 1?1304 9 and 10595 2?1311 5 vs 11157 7?1300 3; t =10 31 and 4 68, P =0 0001 and 0 0054). Furthermore, the inhibition ratios of collagenⅠwas 30 7%, and the one of collagen Ⅲ 5 1%, in the experimental group. Conclusions The expression of collagenⅠand Ⅲ of keloid fibroblasts in vitro may be significantly more intensive than the one of normal skin fibroblasts. It seems that the expression of collagenⅠand Ⅲ, especially collagenⅠ, in keloid fibroblasts and normal skin fibroblasts in vitro may be suppressed greatly by the application of Xiao Ban Xu.