Objective To investigate the cause of excessive collagen accumulation in keloid tissue. Methods The ultrastructure of keloid was observed by transmission electron microscope. New formed collagen in keloid was localized with ABC immunohistochemical staining. Type I procollagen mRNA level in keloid tissue was determined by dot blot hybridization using human pro-al (I)collagenspecific cDNA probe. Results Numerous fibroblasts with abundant, well developed rough endoplasmic reticulum were exhibited in the ultrastructure of keloid. The type I procollagen mRNA levels were significantly increased in kreloid tissue. Immunohistochemical staining showed increased expression of new formed, type I procollagen in keloid tissue. Conclusion the fibroblasts are activated in collagen synthesis in active keloid. The enhanced collagen synthesis by fibroblasts is a critical factor leading to the overabundant collagen accumulation in keloid.

Objective To explore the effect and mechanism of fluvastatin preventing and curing restenosis. Methods 48 rabbits were randomly divided into 3 groups. In control group, rabbits were given basic food. In balloon injury group, rabbits were given basic food and balloon injury of general artery on right neck. In fluvastatin balloon group, rabbits were given basic food,ballon injury of general artery and 10mg?kg -1 ?d -1 fluvastatin. The expression of MMP-9 and TIMP-2 of mRNA was detected at 3, 7, 14 and 30 days after injury respectively by method of in site hybridization. Results There was little expression of MMP-9 and TIMP-2 mRNA in control group, and the expression of MMP-9 and TIMP-2 mRNA in middle membrane of blood vessels began at 3 days after blood injury, and reached maximum at 7 days after injury. There was a little expression of MMP-9 and TIMP mRNA in inner membrane of blood vessels at 14 and 30 days after injury, and MMP-9 expression significantly decreased after the fluvastatin interference(P

Objective To study the expression and distribution of protein STAT3 in the retina after optic nerve transection. Methods Immunocytochemistry, Western blot and computer image analysis techniques were used. Results After optic nerve transection, STAT3 levels in the retina was highly up-regulated, the peak of which appeared at day 1 postaxotomy, then decreased gradually to the normal level 5 days later. More translocations of STAT3 to the nucleus were seen.Conclusion JAK-STAT signal transduction pathway was involved in the pathological processes in the retinal after transection of the optic nerve.

Objective To investigate the effect of hypoxia-inducible factor (HIF)-1α and vascular en-dothelial growth factor (VEGF) on the angioge-nesis of rheumatoid arthritis (RA). Methods The collagen-induced arthritis (CIA) model was set up in male Wistar rats. The pathological angiogenesis and expression of HIF-1α and VEGF in the synovia different time points were observed by H&E and immunohistochemistry staining. Results The expressions of HIF-1α and VEGF on CIA synovium were significantly elevated and their expression increased gradually with the prolonged disease course. Both synovial HIF-1α expression and VEGF expression were correlated significantly with the pathological angiogenesis score. Synovial lining and sublining HIF-1α expression were correlated significantly with VEGF expression respectively. Conclusion H1F-1α may play an important role in the pathogenesis of RA by upregulating the expression of VEGF and then promoting angiogenesis.

Objective　To investigate the actions of extra cellular medium in growth and differentiation of hair follicle and to look for growth adjusting factors for dermal papilla cells (DPC). Methods　Dermal papilla cells were isolated and cultivated with two steps method and the cells were identified by immunohistochemical staining for actin. Influence was examined on the adhesion and growth of dermal papilla cells by chondroitin sulfate A, chondroitin sulfate C and heparin sulfate. Results　Two steps method of enzyme digestion for isolating and cultivating dermal papilla cells was an efficient method and large amount of dermal papilla of high purity were harvested with this method. The method is very simple and easy to manege with. Increased adhesion and growth of dermal papilla cells were observed in specimen treated with chondroitin A and heparin sulfate. No significant effects was observed in the cells treated with chondroit in sulfate C. Conclusion　Some extra cellular medium can regulate the adhesion and growth of dermal papilla cells and therefore influence the growth and development of hair follicle.

Objective To construct the KU80 inhibition cell model by RNAi in U2OS cell and to explore the relationship between the Ku80,telomeres and radiosensitivity in telomerase-negative tumor cells.Methods U2OS cells were transfected with the recombinant plasmids of pshRNA-K80 by the lipofectamine,and the stable transfected cell clones were selected by G418.After the selection,the cells were collected and analyzed by the flow cytometry.RT-PCR and Western blot were used to measure the expression of Ku80 and Real-time PCR was used to detect the length of telomeres.The radiosensitivity of U2OS was determined by clone formation array.Results The transfection efficiency of the positive cell clones detected by the flow cytometry was (83.23 ± 7.63) ％.The inhibition rate of the Ku80 gene transcription in the cell group with recombinant plasmid was(68.09 ± 1.16)％ and the inhibition rate of the Ku80 protein expression in the same group was (11.03 ± 2.45) ％.The results of Real-time PCR showed that the telomere length of the cell group with recombinant plasmid (1.07 ± 0.07) was significantly shorter than that of the control group (4.42 ± 1.30,F =38.58,P ＜ 0.05) and that of the empty plasmid group (4.11 ±0.84,F =38.58,P ＜ 0.05).Compared to the control group,the telomere length of the empty plasmid group did not changed(4.42 ±0.84 vs.4.11 ±0.84).U2OS cells with Ku80 expression suppressed had lower SF2 than that of the control cells (F =1089.61,P ＜0.05),and resulted in the SER of 1.47.Conclusions The Ku80 inhibition cell model in telomerase-negative U2OS cell line is successfully established which has the shorter telomere length,and is more sensitive to radiation.Telomere shortening caused by pshRNA-of Ku80 is likely to be one of the mechanisms of radiosensitization in this kind of cell model.

Objective To investigate the distribution of elderly metabolic syndrome(MS)and its components in the elderly population,and to analyze its correlation with the common chronic diseases in the elderly.Methods The general situation,history of diseases and health examination results were collected in 362 elderly with an average age of 79.2 years visiting Beijing Hospital.They were grouped into 3 groups of the young old age(65-74 years old),the middle old age(75-84 years old)and the older old age(85-98 years).The prevalence distribution of MS and its correlation with the common chronic diseases in the elderly were analyzed.Results MS prevalence in average was 28.4％ (103 cases)in the elderly population,with 18.6％ (67 cases) in the young old group,26.3％ (95 cases)in middle old group,and 33.7％ (122 cases)in older old group.The tendency was rising up with age increase,and the MS prevalence was higher in older old group than in other groups (x2 =8.57,P =0.03).Correlation was observed between genesis of MS and common geriatric chronic diseases.The prevalence differences in hypertension,prostatic hyperplasia,diabetes mellitus,and coronary heart disease were statistically significant higher in MS group than in non-MS group(P =0.02,0.01,0.02,0.03,respectively).Conclusions MS prevalence of elderly population is relatively high,and has significant correlation with other chronic diseases.We need to recognize the harm of MS,to conduct regular monitoring and management according to risk level,and practice patient educations to reduce MS negative effects on elderly life quality and health.

Objective To screen and clone the genes related to alopecia areata. Methods Dermal papillae of lesional and non lesional follicles were separated from alopecia areata scalp respectively. Suppression subtractive hybridization was used to investigate the difference of expressed genes in dermal papillae of lesional (tester) and non lesional (driver) follicles, and the differentially expressed genes in dermal papillae of lesional follicles were cloned and sequenced. Results A subtractive library of dermal papillae of lesional follicles from alopecia areata was established. A differentially expressed gene in dermal papillae of lesional follicles was successfully cloned and proved to be an autoantigen gene. Conclusions The subtractive library may contain the differentially genes related to alopecia. The autoantigen gene related to alopecia areata need to be further investigated.

Female ; Humans ; Middle Aged ; Postmenopause ; Trophoblastic Neoplasms ; pathology ; Uterine Neoplasms ; pathology

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.