Objective To investigate the effects of asiaticoside onthe proliferation and the Smad signal pathway of the hypertrophic scar fibroblasts.Methods The hypertrophic scar fibroblasts were cultured with tissue culture method. The expressions of Smad2 and Smad7 mRNA after asiaticoside treatment were determined by reverse transcriptionpolymerase chain reaction 48 hours later. Thecell cycle, the cell proliferation, the cell apoptosis and the expression of phosphorylated Smad2 and Smad7 with(experimental group) or without(control group) asiaticoside were detected with flow cytometry, immunocytochemistry and Western blot. Results Asiaticoside inhibited the hypertrophic scar fibroblasts from phase S to phase M. The Smad7 content and the expression of Smad7 mRNA were (1.33±1.26)% and (50.80±22.40)% in experimental group, and (9.15±3.36)% and (32.18±17.84)% in control group; there were significant differences between two groups (P＜0.05). While the content and the mRNA expression of Smad2 had no significant difference between two groups. Conclusion Asiaticoside inhibits the scar formation through Smad signal pathway.
Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide （PIK3cb） shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups （n=25, in each group）: blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 （shRNA-1+shRNA-2）, negative control (pGenesil-1 scramble shRNA) and positive control （wortmannin） group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P＜0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P＜0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P＜0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
【摘要】目的探讨腹膜后血管滤泡增生症的临床特点及诊治方法。方法对我院收治的3例病例并结合相关文献做一回顾性分析。结果该疾病主要临床表现为腹部包块、贫血等症状，均经手术治疗。术中见包块直径为7～18 cm。2例手术完整切除全块，其中1例病理检查为透明血管型，随访至今（7个月）健在，无复发； 另1例为浆细胞型，随访2年后失访。余1例仅行剖腹探查术，取部分组织作病理检查，病理报告为浆细胞型，术后8个月死亡。结论本病缺乏特异性的临床表现和影像特点，术前难以诊断，确诊须依靠病理检查。手术完整切除肿瘤是治疗本病的最佳方法。
Objective To explore the change of gene expression of stress activated protein kinase (SAPK) and its upstream signalregulated molecule ——mitogen activated protein kinases(MAPKs) (MKK4 and MKK7) in hypertrophic scar and autocontrol normal skin. Methods The total RNA was isolated from 8 hypertrophic scars and 8 auto-control skin, and then mRNA was purified. The gene expressions of MKK4, MKK7 and SAPK were examined with reverse transcriptionpolymerase chain reaction(RT-PCR) method. Results In hypertrophic scar, both MKK7 and SAPK genes weakly expressed. In auto-control skin, the expression of these 2 genes was significantly elevated in comparison with hypertrophic scar (Plt;0.01). The expression levelsof these 2 genes were 1.5 times and 2.6 times as long as those of hypertrophic scar, respectively. Gene expression of MKK4 had no significant difference between autocontrol skin and hypertrophic scar (Pgt;0.05). Conclusion Decreased gene expression of MKK7 and SAPK which results in reducing cell apoptosis might be one of the mechanisms for controlling the formation of hypertrophic scar.
Objective To investigate the relationship between keloid proliferation and destruction of skin appendages(SAs). Methods Pathological biopsies of keloids were derived from 17 patients whounderwent scar resection. All samples were divided into 4 groups: infiltrating growth locus of keloids（K-I,n=9），proliferative keloids (K-P,n=17), atrophic keloids (K-A,n=10), and edging normal skin (K-N,n=6). Normal skin derived from thorax of patients was used as control (NS, n=6). The density of SAs and the expressive characteristics of pan-cytokeratin (CKp), cytokeratin19 (CK19), secretory component of glandular epithelium(SC), proliferating cell nuclear antigen(PCNA), and apoptosis related proteins (Bcl-2 and Bax) were observed with immunohistochemical method. Results Compared with K-N and NS, the density of SAs expressing CKP and SC in keloids was apparently decreased, and remnant of CKp protein was observed after the disappearance of SAs structures. Protein expression of Bax was increased in epithelial cellsof most SAs. SAs containing postive immunostaining signals of Bcl-2, PCNA and CK19 exhibited squamous epithelization and abnormal structure. The structure of SAs underwent 3 morphological stages: infiltrating, proliferating, and maturing.In correspondence to each stage, SAs underwent proliferation, structural destruction, and fibrosis which were caused by cellular migration, nflammatory reaction, and vascular occlusion respectively. Conclusion Abnormal proliferation of epithelial cells and their structural destruction of SAs may beassociated with tissue fibrosis in keloid lesion.
Objective To analyze the clinical risk factors of the occurrence of severe proliferative vitreoretinopathy (PVR) after scleral reattachment surgery. Methods A total of 4031 eyes of 4031 consecutive patients with reghmatogenous retinal detachment (RRD) and PVR (grade C1 or less), on whom the scleral buckling was performed, were retrospectively studied. Twenty-two clinical charac teristics of the patients (including the ocular tension, condition of lens and vitreous, characte ristics of retinal detachment, whether or not with choroidal detachment, et al) were recorded.In 4031 patients, 2660 were followed up for more than 3 months, and 72 (in PVR group) of the 2660 patients underwent the second surgery (vitre oretinal surgery) because of the occurrence of postoperative seve re PVR; in the other 2588 patients, 72 (72 eyes) with retinal reattachment for more than 3 months were selected randomly as the control. The data were analyzed in SPSS (10.0) software. Results Logistic regression analysis revealed that the significant risk factors for PVR were incomplete posterior vitreous detachment ( P＜0.001), intraocular pressure lt;7 mm Hg（1 mm Hg=0.133 kPa, P＜0.002）, and large retinal tear (gt;2 DD,P＜0.005). Conclusion Incomplete posterior vitreous detachment, intraocular pressure lt;7 mm Hg and large retinal tear of the patient with RRD may be the major risk factors for PVR. (Chin J Ocul Fundus Dis，2003，19：141-143)
Objective To evaluate the effect of vascular endothelial cell growth factor (VEGF) antisense oligodeoxynucleotides (ASODNs) on the expression of VEGF in rats with oxygen-induced retinopathy. Methods Thirty newborn Sprague-Dawley (SD) rats were randomly divided into 3 groups:normal control group, disposal group and non-disposed group, The animal models with oxygen-induced proliferative retinopathy were established by raising the rats in hyperoxic environment. Retrobulbar injection was performed with VEGF ASODNs or normal saline on the rats in 3 groups respectively. The intraocular tissues (all the tissues except the cornea, sclera, and lens) and serum were collected, and the expressions of VEGF were determined by using competitive enzyme immunoassay.Results The expressions of VEGF in intraocular tissues of rats in disposal group were significantly lower than those in non-disposed group (P＜0.05), and there was no statistical difference between the disposal and normal control group (P＞0.05). There was no significant difference of the expressions of VEGF in serum of rats between the disposal and non-disposed group (P＞0.05), which were both lower than those in the normal control group (P＜0.05). Conclusion VEGF ASODNs could significantly inhibit the expression of VEGF in intraocular tissues. (Chin J Ocul Fundus Dis，2003，19：172-174)