Uterine Fibroid: Isolation, Culture and Characterization

Present study was carried out to establish uterine fibroid primary culturesystem for screening of natural/synthetic compounds against uterine fibroid. For invitro culture, enzymatic isolation method was used. To characterize, histochemistry (H& E, Masson’s Trichrome and Periodic Acid Schiff) staining and immunocytochemistryusing marker antibodies (Versican) were performed in vitro. Uterine fibroid tissueshowed much intense staining of Masson’s Trichrome and Periodic Acid Schiff stain ascompared to adjacent myometrium tissue. The primary cultured cells showedsignificantly higher proliferation, sub-culture efficiency and expression of Versicanprotein. In conclusion, our results suggest that in vitro cultured uterine fibroid cells mayoffer a suitable alternative model to evaluate natural or synthetic compounds havingantitumor properties for uterine fibroid treatment.

Expressions of Versican and ADAMTS-1 in polycysticovary syndrome patients and their clincal significances / 吉林大学学报(医学版)

Objective: To determine the expressions of Versican and a distintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1) in serum in the polycystic ovary syndrome (PCOS) patients, and to clarify their roles in the pathogenesis of PCOS. Methods: A total of 80 patients with PCOS (PCOS group) and 100 healthy women (control group) were selected The heights and body weights of the subjects in two groups were measured; and the body mass index (BMI) was calculated The levels of serum Versican and ADAMTS-1 of the subjects in two groups were measured by enzyme-linked immunosorbent assay (ELISA) method The levels of serum follicle stimulating hormone (FSH), luteinizing hormone (L H), testosterone (T), fasting blood glucose (FBG), fasting insulin of the subjects in two groups were detected; insulin resistance was evaluated according to Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). The correlations between Versican, ADAMTS-1 and the metabolic indexes were analyzed by Pearson linear correlation analysis Results: The serum Versican level of the patients in PCOS group was significantly decreased compared with control group (P = 0 . 004); the serum ADAMTS-1 level was also decreased (P < 0 . 01). The sensitivities, the specificities and the area under receiver operating characteristic (ROC) curve (AUC) of Versican and ADAMTS-1 in prediction of PCOS were 76.74% vs 63. 64% (P = 0 . 018), 52.94% 1)5 40.73% (P = 0 . 009) and 0. 675 (0.550-0.795) vs 0.714 (0 . 6 0 1 - 0. 827) (P = 0 . 032), respectively. The correlation analysis showed that in PCOS group the serum level of Versican of the patients was negatively correlated with FBG (r = - 0 . 7 3 8, P= 0.022), and ADAMTS-1 was negatively correlated with FBG (r = -0.524, P = 0.043). Conclusion: Versican and ADAMTS-1 lowly express in the patients with PCOS. They are negatively correlated with the insulin resistance. They may play inhibitory effects in the occurrence of PCOS.

Expressions of Versican and ADAMTS-1 in polycysticovary syndrome patients and their clincal significances / 吉林大学学报(医学版)

Objective: To determine the expressions of Versican and a distintegrin and metalloprotease with thrombospondin type l motifs (ADAMTS-1) in serum in the polycystic ovary syndrome (PCOS) patients, and to clarify their roles in the pathogenesis of PCOS. Methods: A total of 80 patients with PCOS ( PCOS group) and 100 healthy women (control group) were selected. The heights and body weights of the subjects in two groups were measured; and the body mass index (BMI) was calculated. The levels of serum Versican and ADAMTS-1 of the subjects in two groups were measured by enzyme-linked immunosorbent assay (ELISA) method. The levels of serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), fasting blood glucose (FBG), fasting insulin of the subjects in two groups were detected; insulin resistance was evaluated according to Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). The correlations between Versican, ADAMTS-1 and the metabolic indexes were analyzed by Pearson linear correlation analysis. Results: The serum Versican level of the patients in PCOS group was significantly decreased compared with control group (P=0.004); the serum ADAMTS-1 level was also decreased (P<0.01). The sensitivities, the specificities and the area under receiver operating characteristic (ROC) curve （AUC）of Versican and ADAMTS-1 in prediction of PCOS were 76.74%?vs 63.64% (P=0.018), 52.94% vs 40.73% (P=0.009) and 0675 (0.550-0.795) vs 0.714 (0.601-0.827) (P=0.032), respectively. The correlation analysis showed that in PCOS group the serum level of Versican of the patients was negatively correlated with FBG (r=-0.738, P=0.022), and ADAMTS-1 was negatively correlated with FBG (r=-0.524, P=0.043). Conclusion； Versican and ADAMTS-1 lowly express in the patients with PCOS. They are negatively correlated with the insulin resistance. They may play inhibitory effects in the occurrence of PCOS.

Expression of ADAMTS4 in the heart of myocardial infarction rat / 国际生物医学工程杂志

Objective To detect the expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4),Versican,and Aggrecan in hearts of myocardial infarction rats.Methods Male Wistar rats were chosen to make myocardial infarction model.Fifteen rats were randomly assigned into the myocardial infarction group and five rats into the sham-operation group.Rats were killed 3 d after surgery and the left ventricles were taken to produce frozen section.ADAMTS4,Versican,and Aggrecan protein expressions were detected by imnunohistochemistry method.Results ADAMTS4 expressed in myocardial cells at marginal zone of the infarcted area.ADAMTS4 expressed weakly while Versican expressed strongly in capillary endothelial cells of marginal zone of the infarcted area.Aggrecan showed no expression in the cardiac tissue and vascular endothelium of both myocardial infarction group and sham-operation group.Conclusions ADAMTS4 is involved in the inflammation after myocardial infarction.Degradation of ADAMTS4 to Versican exists in the vessels after myocardial infarction.Aggrecan is not involved in the pathophysiological process of myocardial infarction.

Progress of versican in the occurrence and development of neoplasms / 国际外科学杂志

Versican comes from fibroblasts and belongs to chondroitin sulfate proteoglycan.It distributes in various tissues.The versican have functions including regulate cell growth and differentiation,promote cell proliferation,invasion and metastasis,and stimulate angiogenesis.Experiments have confirmed that the expression of versican is involved in various cancers.Here,the author do an overview of versican in the occurrence and development of breast cancer,liver cancer and chondrosarcoma aimed to further explore the pathogenesis of tumors.

Influencia de componentes de la matriz extracelular en la agresividad y potencial invasor del cáncer cérvicouterino: ¿Qué hay de nuevo? / Influence of extracellular matrix components in aggressiveness and invasive potential of cervical cancer: What's new?

El cáncer cérvico uterino (CCU) es una patología de alta incidencia y mortalidad. La investigación hasta ahora se ha enfocado en estudiar su asociación con virus papiloma. Sin embargo, el estudio de la matriz extracelular (MEC) ha dado una nueva perspectiva para el estudio de factores inductores o perpetuadores de las neoplasias. En las neoplasias epiteliales como CCU el estroma tumoral presenta una composición dinámica de elementos celulares, destacando la presencia de miofibroblastos positivos a alfa actina de músculo liso (alfa SMA+) y fibrocitos CD34+. La MEC tiene un papel fundamental, ya que no sólo otorga las condiciones apropiadas para el desarrollo del tumor, sino que además condiciona el fenotipo de la población celular del estroma, donde la pérdida de fibrocitos CD34+ asociada a una ganancia de miofibroblastos alfa SMA+ podría ser un indicador muy sensible de invasión estromal, incluso en estadios iniciales. De la misma forma lo hace TGF-beta Ι, ya que su presencia es un reflejo de la síntesis de alfa SMA. Un nuevo elemento es versicán, un proteoglicano cuyas isoformas V0 y V1 se expresan también en tejidos neoplásicos de tumores ováricos, mama y cerebro, entre otros. Desempeña un papel muy importante en los fenómenos de adhesión celular, proliferación, migración y ensamblaje a la MEC. Por lo tanto, el análisis del estroma adyacente a las lesiones epiteliales del cuello uterino puede complementar el conocimiento sobre la conducta biológica de éstas, constituyendo una poderosa herramienta diagnóstica, de forma complementaria a los elementos utilizados hasta ahora.

Squamous cell carcinoma of the cervix (SCC) is a pathology that has high incidence and mortality. So far, research has been focused in the study of its association with papilloma virus. However, knowledge about extracellular matrix (ECM) has given a new perspective for the study of factors that induce or perpetuate neoplasms. In epithelial neoplasms like SCC, the tumoral stroma exhibits a dynamic composition of cellular elements, highlighting the presence of alpha actin of smooth muscle positive myofibroblasts (alpha SMA+) and CD34+ fibrocytes. ECM has an essential role, because it not only provides the appropriate conditions for tumor's development, but also affects stromal cell population phenotype, where a loss of CD34+ fibrocytes associated with a gain of alpha SMA+ myofibroblasts could be a sensitive indicator of stromal invasion, even in early stages. TGF-beta Ι does it in the same way, as its presence is a reflection of the synthesis of alpha SMA+. A new element is versican, a proteoglycan whose V0 and V1 isoforms expression is also observed in neoplastic tissues of ovary, breast and brain tumors, among others. It plays an important role in the phenomena of cellular adhesion, proliferation, migration and assembly of the ECM. Therefore, the analysis of the stroma adjacent to epithelial injuries of the cervix can complement the knowledge about the biological conducts of these, constituting a powerful diagnostic tool, as a complement to the elements used nowadays.

The Effects of TWEAK, Fn14, and TGF-beta1 on Degeneration of Human Intervertebral Disc

OBJECTIVE: The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-beta1 (TGF-beta1) on degeneration of human intervertebral disc (IVD). METHODS: Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in 37degrees C, 5% CO2 incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-beta1, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-beta1 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-beta1 but the result was insignificant statistically. TGF-beta1 increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-beta1 in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican. CONCLUSION: This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.

Induction of spondylitis and sacroiliitis in mice by the G1 domain of the proteoglycan versican / 解放军医学杂志

Objective To set up rodent ankylosing spondylitis model. Methods Recombinant natively folded versican G1 domain(VG1) and FCA were injected into BALB/c mice to induce spondylitis and sacroiliitis. Spondylitis and sacroiliitis were determined by pathological examination. Results It was shown that immunity to recombinant VG1 resulted in spondylitis in the lumbar spine and sacroiliitis in 35 0% and 12 5% mice respectively. Accumulation of mononuclear cells was observed in spinal ligaments adjacent to the intervertebral disc, the intervertebral disc and the sacroiliac joints. No clinical peripheral arthritis was observed. Conclusion These observations suggest that immunity to human VG1 is involved in the induction of experimental spondylitis and sacroiliitis in BALB/c mice, which will do some help to probe the pathogenesis of human ankylosing spondylitis