Morphologic Variations of the Umbilical Ring, Umbilical Ligaments and Ligamentum Teres Hepatis

Purpose

The varied morphology of the umbilical ring and its surrounding structures, such as the ligamentum teres hepatis, and the median and medial umbilical ligaments, has not been thoroughly investigated. Hence, this study was undertaken to clarify the morphologic variations of these structures.

Materials and Methods

The anterior abdominal walls were removed en bloc from 57 adult cadavers and dissected under a surgical microscope.

Results

One case of umbilical hernia was observed, and the remaining 56 umbilical rings were classified into 3 types: oval or round in 33 cases (Type A, 59.0%), obliterated or slitted in 12 cases (Type B, 21.4%), and completely covered by a connecting band between the ligamentum teres hepatis and umbilical ligaments in 11 cases (Type C, 19.6%). The median and medial umbilical ligaments were classified into four types based on their interrelationships. The most common type was the median umbilical ligament terminated by joining one or both medial umbilical ligaments (Type II, 41.1%). The ligamentum teres hepatis frequently ended by dividing into several branches in the area cranial to the umbilical ring, some of which crossed the umbilical ring. The umbilical fascia covered the umbilical ring in 50.0% of cases, and the rest either not covering the ring or not existing.

Conclusion

These results are expected to improve our understanding of the anatomy of the umbilical area, and further improve treatments of the umbilical hernia.     

Comparison of magnetic resonance imaging and computed tomography-myelography for quantitative evaluation of lumbar intracanalar cross-section

Purpose

A comparison of MRI and computed tomography-myelography (CTM) for lumbar intracanalar dimensions. To compare the capability and reproducibility of MRI and CTM in measuring the cross-sectional morphology of intracanalar lesions of the lumbar spine.

Materials and Methods

MRI and CTM of lumbar disc levels from 61 subjects with various lumbar spinal diseases were studied. Dural area, dural anteroposterior (AP) diameter, dural right-left diameter, and thickness of the ligamentum flavum were measured by two orthopedic surgeons. Each section was graded by degree of stenosis. Absolute value and intra- and inter-observer correlation coefficients (ICC) of these measurements and the associations between MRI and CTM values were determined.

Results

Except for MRI determination of ligament flavum thickness, CTM and MRI and intra- and ICC suggested sufficient reproducibility. When measurements of dural area, dural AP diameter, and RL diameter were compared, values in CTM were significantly (p = 0.01-0.004) larger than those in MRI (CTM/MRI ratios, 119%, 111%, and 105%, respectively). As spinal stenosis became more severe, discrepancies between CTM and MRI in measurements of the dural sac became larger.

Conclusion

Both CTM and MRI provided reproducible measurements of lumbar intracanalar dimensions. However, flavum thickness may be more accurately measured by CTM. Because the differences in the measurements between CTM and MRI are very slight and there is very little data to suggest that the precise degree of stenosis is related to symptoms or treatment outcome, the usefulness of the CTM over MRI needs to be confirmed in future studies.     

胸椎黄韧带骨化症与骨形态发生蛋白4基因单核苷酸多态性的关联

背景：骨形态发生蛋白4基因与胸椎黄韧带骨化症的相关性研究目前较少。 目的：测观察骨形态发生蛋白4的2个单核苷酸多态性位点rs17563和rs2855532与胸椎黄韧带骨化症的关联。 方法：胸椎黄韧带骨化症患者和正常对照者各40例,收集受试者的周围静脉血提取DNA,PCR法进行目的片段骨形态发生蛋白4的单核酸多态性位点rs17563和rs2855532的扩增并测序。 结果与结论：黄韧带骨化症组中rs17563和rs2855532位点带“T”基因型及等位基因型频率明显高于对照组(P < 0.05)。证实,骨形态发生蛋白4上的2个单核酸多态性位点rs17563和rs2855532等位基因型突变与胸椎黄韧带骨化症的发生有关。     

Characterization of human adipose-derived stem cells and expression of chondrogenic genes during induction of cartilage differentiation

OBJECTIVES:

Understanding the changes in chondrogenic gene expression that are involved in the differentiation of human adipose-derived stem cells to chondrogenic cells is important prior to using this approach for cartilage repair. The aims of the study were to characterize human adipose-derived stem cells and to examine chondrogenic gene expression after one, two, and three weeks of induction.

MATERIALS AND METHODS:

Human adipose-derived stem cells at passage 4 were evaluated by flow cytometry to examine the expression of surface markers. These adipose-derived stem cells were tested for adipogenic and osteogenic differentiation capacity. Ribonucleic acid was extracted from the cells for quantitative polymerase chain reaction analysis to determine the expression levels of chondrogenic genes after chondrogenic induction.

RESULTS:

Human adipose-derived stem cells were strongly positive for the mesenchymal markers CD90, CD73, CD44, CD9, and histocompatibility antigen and successfully differentiated into adipogenic and osteogenic lineages. The human adipose-derived stem cells aggregated and formed a dense matrix after chondrogenic induction. The expression of chondrogenic genes (collagen type II, aggrecan core protein, collagen type XI, COMP, and ELASTIN) was significantly higher after the first week of induction. However, a significantly elevated expression of collagen type X was observed after three weeks of chondrogenic induction.

CONCLUSION:

Human adipose-derived stem cells retain stem cell characteristics after expansion in culture to passage 4 and serve as a feasible source of cells for cartilage regeneration. Chondrogenesis in human adipose-derived stem cells was most prominent after one week of chondrogenic induction.     

Total absence of the alpha2(I) chain of collagen type I causes a rare form of Ehlers-Danlos syndrome with hypermobility and propensity to cardiac valvular problems

Background

Heterozygous mutations in the COL1A1 or COL1A2 gene encoding the α1 and α2 chain of type I collagen generally cause either osteogenesis imperfecta or the arthrochalasis form of Ehlers‐Danlos syndrome (EDS). Homozygous or compound heterozygous COL1A2 mutations resulting in complete deficiency of the proα2(I) collagen chains are extremely rare and have been reported in only a few patients, albeit with variable phenotypic outcome.

Methods

The clinical features of the proband, a 6 year old boy, were recorded. Analysis of proα and α‐collagen chains was performed by SDS‐polyacrylamide gel electrophoresis using the Laemmli buffer system. Single stranded conformation polymorphism analysis of the proband''s DNA was also carried out.

Results

In this report we show that complete lack of proα2(I) collagen chains can present as a phenotype reminiscent of mild hypermobility EDS during childhood.

Conclusions

Biochemical analysis of collagens extracted from skin fibroblasts is a powerful tool to detect the subset of patients with complete absence of proα2(I) collagen chains, and in these patients, careful cardiac follow up with ultrasonography is highly recommended because of the risk for cardiac valvular problems in adulthood.     

Sex,Collagen Expression,and Anterior Cruciate Ligament Strength in Rats

Context:

Sex-specific responses to steroid sex hormones have been suggested as a potential cause for the disparate anterior cruciate ligament (ACL) injury rates between male and female athletes. Type 1 collagen (T1C) and type 3 collagen (T3C) are crucial structural components that define the ligament''s ability to withstand tensile loads. Messenger RNA (mRNA) is an important mediator of downstream collagen synthesis and remodeling, but the sex-specific mechanisms of collagen mRNA expression and ACL strength are unknown.

Objective:

To examine the influence of sex on T1C and T3C mRNA expression and mass-normalized stiffness and peak failure load in the ACLs of skeletally mature rats.

Design:

Observational study.

Setting:

Basic sciences and biomechanical testing laboratories.

Patients or Other Participants:

Nineteen 12-week-old male (n  =  9) and female (n  =  10) Sprague Dawley rats.

Main Outcome Measure(s):

We used real-time polymerase chain reaction to determine T1C and T3C mRNA expression and a hydraulic materials testing device to measure ACL stiffness and failure load. Nonparametric Wilcoxon rank sum tests were used to compare the groups.

Results:

Female rats had lower amounts of T3C mRNA expression and higher normalized ACL tangent stiffness and failure load than male rats.

Conclusions:

These findings suggest that sex-specific differences in T1C and T3C mRNA expression may play an important role in the downstream mechanical properties of the ACL.     

Upregulation of matrix synthesis in chondrocyte-seeded agarose following sustained bi-axial cyclic loading

OBJECTIVES:

The promotion of extracellular matrix synthesis by chondrocytes is a requisite part of an effective cartilage tissue engineering strategy. The aim of this in vitro study was to determine the effect of bi-axial cyclic mechanical loading on cell proliferation and the synthesis of glycosaminoglycans by chondrocytes in three-dimensional cultures.

METHOD:

A strain comprising 10% direct compression and 1% compressive shear was applied to bovine chondrocytes seeded in an agarose gel during two 12-hour conditioning periods separated by a 12-hour resting period.

RESULTS:

The bi-axial-loaded chondrocytes demonstrated a significant increase in glycosaminoglycan synthesis compared with samples exposed to uni-axial or no loading over the same period (p<0.05). The use of a free-swelling recovery period prior to the loading regime resulted in additional glycosaminoglycan production and a significant increase in DNA content (p<0.05), indicating cell proliferation.

CONCLUSIONS:

These results demonstrate that the use of a bi-axial loading regime results in increased matrix production compared with uni-axial loading.     

Petroleum Ether Extract of Cissus Quadrangularis (Linn.) Enhances Bone Marrow Mesenchymal Stem Cell Proliferation and Facilitates Osteoblastogenesis

OBJECTIVE

To evaluate the effects of the petroleum ether extract of Cissus quadrangularis on the proliferation rate of bone marrow mesenchymal stem cells, the differentiation of marrow mesenchymal stem cells into osteoblasts (osteoblastogenesis) and extracellular matrix calcification. This study also aimed to determine the additive effect of osteogenic media and Cissus quadrangularis on proliferation, differentiation and calcification.

METHODS

MSCs were cultured in media with or without Cissus quadrangularis for 4 weeks and were then stained for alkaline phosphatase. Extracellular matrix calcification was confirmed by Von Kossa staining. marrow mesenchymal stem cells cultures in control media and osteogenic media supplemented with Cissus quadrangularis extract (100, 200, 300 μg/mL) were also subjected to a cell proliferation assay (MTT).

RESULTS

Treatment with 100, 200 or 300 μg/mL petroleum ether extract of Cissus quadrangularis enhanced the differentiation of marrow mesenchymal stem cells into ALP-positive osteoblasts and increased extracellular matrix calcification. Treatment with 300 μg/mL petroleum ether extract of Cissus quadrangularis also enhanced the proliferation rate of the marrow mesenchymal stem cells. Cells grown in osteogenic media containing Cissus quadrangularis exhibited higher proliferation, differentiation and calcification rates than did control cells.

CONCLUSION

The results suggest that Cissus quadrangularis stimulates osteoblastogenesis and can be used as preventive/ alternative natural medicine for bone diseases such as osteoporosis.     

Lung Morphometry,Collagen and Elastin Content: Changes after Hyperoxic Exposure in Preterm Rabbits

INTRODUCTION

Elastic and collagen fiber deposition increases throughout normal lung development, and this fiber network significantly changes when development of the lung is disturbed. In preterm rats and lambs, prolonged hyperoxic exposure is associated with impaired alveolization and causes significant changes in the deposition and structure of elastic fibers.

OBJECTIVES

To evaluate the effects of hyperoxic exposure on elastic and collagen fiber deposition in the lung interstitial matrix and in alveolarization in preterm rabbits.

METHODS

After c-section, 28-day preterm New-Zealand-White rabbits were randomized into 2 study groups, according to the oxygen exposure, namely: Room air (oxygen = 21%) or Oxygen (oxygen ≥ 95%). The animals were killed on day 11 and their lungs were analyzed for the alveolar size (Lm), the internal surface area (ISA), the alveoli number, and the density and distribution of collagen and elastic fibers.

RESULTS

An increase in the Lm and a decrease in the alveoli number were observed among rabbits that were exposed to hyperoxia with no differences regarding the ISA. No difference in the density of elastic fibers was observed after oxygen exposure, however there were fewer collagen fibers and an evident disorganization of fiber deposition.

DISCUSSION

This model reproduces anatomo-pathological injuries representing the arrest of normal alveolar development and lung architecture disorganization by just a prolonged exposition to oxygen.

CONCLUSIONS

In the preterm rabbit, prolonged oxygen exposure impaired alveolization and also lowered the proportion of collagen fibers, with an evident fiber network disorganization.     

Elevated levels of dehydroepiandrosterone as a potential mechanism of dendritic cell impairment during pregnancy

Background

This study aimed to test the hypothesis that immune dysfunction and the increased risk of spontaneous abortion in pregnant women with hyperandrogenia (HA) are caused by the reduced tolerogenic potential of dendritic cells (DCs) that results from elevated levels of dehydroepiandrosterone sulfate (DHEAS).

Methods

The phenotypic and functional properties of monocyte-derived DCs generated from blood monocytes from non-pregnant women, women with a normal pregnancy, or pregnant women with HA, as well as the in vitro effects of DHEAS on DCs in healthy pregnant women were investigated.

Results

In a normal pregnancy, DCs were shown to be immature and are characterized by a reduced number of CD83⁺ and CD25⁺ DCs, the ability to stimulate type 2 T cell responses and to induce T cell apoptosis. By contrast, DCs from pregnant women with HA had a mature phenotype, were able to stimulate both type 1 (IFN-γ) and type 2 (IL-4) T cell responses, and were characterized by lower B7-H1 expression and cytotoxic activity against CD8⁺ T cells. The addition of DHEAS to cultures of DCs from healthy pregnant women induced the maturation of DCs and increased their ability to activate type 1 T cell responses.

Conclusion

Our data demonstrated the reduction in the tolerogenic potential of DCs from pregnant women with HA, and revealed new mechanisms involved in the hormonal regulation of DCs mediated by DHEAS.     

Redefining the risks of prenatally ascertained supernumerary marker chromosomes: a collaborative study

Background

A marker chromosome is defined as a structurally abnormal chromosome that cannot be identified by routine cytogenetics. The risk for phenotypic abnormalities associated with a marker chromosome depends on several factors, including inheritance, mode of ascertainment, chromosomal origin, and the morphology, content, and structure of the marker.

Methods

to understand the karyotype‐phenotype relationship of prenatally ascertained supernumerary de novo marker chromosomes, we combined data from prenatal cases obtained from 12 laboratories with those from studies in the literature. We were able to obtain cytogenetic and phenotypic data from 108 prenatally ascertained supernumerary de novo marker chromosomes to refine the phenotypic risk associated with these markers. Because of the growing number of cases and because more techniques are available to delineate marker morphology, we have been able to group risk estimates into subcategories, such as by marker type and whether there are ultrasound abnormalities.

Results

If a de novo supernumerary marker chromosome is found prenatally, our data suggest there is a 26% risk for phenotypic abnormality when there is no other information defining the marker (such as chromosomal origin or information about the existing phenotype). However, if high resolution ultrasound studies are normal, this risk reduces to 18%.

Conclusions

Our findings strongly support the value of additional genetic studies for more precisely defining the risk in individual cases involving marker chromosomes.     

Rapamycin Inhibits Transforming Growth Factor β1-Induced Fibrogenesis in Primary Human Lung Fibroblasts

Purpose

The present study was designed to determine whether rapamycin could inhibit transforming growth factor β1 (TGF-β1)-induced fibrogenesis in primary lung fibroblasts, and whether the effect of inhibition would occur through the mammalian target of rapamycin (mTOR) and its downstream p70S6K pathway.

Materials and Methods

Primary normal human lung fibroblasts were obtained from histological normal lung tissue of 3 patients with primary spontaneous pneumothorax. Growth arrested, synchronized fibroblasts were treated with TGF-β1 (10 ng/mL) and different concentrations of rapamycin (0.01, 0.1, 1, 10 ng/mL) for 24 h. We assessed m-TOR, p-mTOR, S6K1, p-S6K1 by Western blot analysis, detected type III collagen and fibronectin secreting by ELISA assay, and determined type III collagen and fibronectin mRNA levels by real-time PCR assay.

Results

Rapamycin significantly reduced TGF-β1-induced type III collagen and fibronectin levels, as well as type III collagen and fibronectin mRNA levels. Furthermore, we also found that TGF-β1-induced mTOR and p70S6K phosphorylation were significantly down-regulated by rapamycin. The mTOR/p70S6K pathway was activated through the TGF-β1-mediated fibrogenic response in primary human lung fibroblasts.

Conclusion

These results indicate that rapamycin effectively suppresses TGF-β1-induced type III collagen and fibronectin levels in primary human lung fibroblasts partly through the mTOR/p70S6K pathway. Rapamycin has a potential value in the treatment of pulmonary fibrosis.     

Hsp70 Knockdown by siRNA Decreased Collagen Production in Keloid Fibroblasts

Purpose

There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive collagen production in keloid fibroblasts.

Materials and Methods

We inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected small interfering RNAs (siRNAs) specific for human Hsp70, transfected them into keloid fibroblasts, and evaluated the resulting phenotypes and protein production using real-time polymerase chain reaction (PCR), Western blot, and a collagen assay.

Results

The siRNAs dramatically suppressed Hsp70 mRNA expression, resulting in a decrease in collagen production in the keloid fibroblasts compared with controls. The siRNAs did not influence the viability of the keloid fibroblasts.

Conclusion

Hsp70 overexpression likely plays an important role in the excessive collagen production by keloid fibroblasts. RNA interference has therapeutic potential for the treatment of keloids.     

PTK2 and PTPN11 expression in myelodysplastic syndromes

OBJECTIVE:

The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes.

METHODS:

Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors.

RESULTS:

Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells.

CONCLUSIONS:

Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors.     

Expression of Glycolysis-Related Proteins in Solid Papillary Carcinoma of the Breast According to Basement Membrane Status

Purpose

The aim of this study was to investigate the differences of expression in glycolysis-related proteins such as Glut-1, carbonic anhydrase (CA) IX, and monocarboxylate transporter (MCT) 4 according to the myoepithelial cell (MEC) and basement membrane (BM) status in solid papillary carcinoma (SPC) of the breast.

Materials and Methods

Immunohistochemical evaluation of Glut-1, CAIX, and MCT4, as well as p63 and type IV collagen, were performed on 23 SPC cases.

Results

Six and nine cases of SPC showed the presence and absence of myoepithelial cells, respectively, and eight cases belonged to the borderline status (p63-positive MEC on some areas of the outer tumor surface but not in others). BM was partially or completely absent in 14 cases and present in nine cases. SPC lacking BM more frequently showed high expression of CAIX than SPC with BM (p=0.037).

Conclusion

In SPC of the breast, a strong expression of CAIX seems to be associated with an increasing degree of loss of BM, which can be interpreted as BM degradation due to the induction of extracellular acidity with increasing expression of CAIX.     

Histomorphometric Analysis of Cutaneous Remodeling in the Early Stage of the Scleroderma Model

INTRODUCTION/OBJECTIVES

Systemic sclerosis, or scleroderma, is a rheumatic disease characterized by autoimmunity, vasculopathy, and fibrosis of the skin and several internal organs. In the present study, our aim was to assess the skin alterations in animals with scleroderma during the first stages of disease induction.

METHODS:

To induce scleroderma, female New Zealand rabbits (n = 12) were subcutaneously immunized with 1 mg/ml of collagen V (Col V) in complete Freund’s adjuvant, twice with a thirty-day interval. Fifteen days later, the animals received an intramuscular booster with type V collagen in incomplete Freund’s adjuvant, twice with a fifteen-day interval. The control group was inoculated with 1 ml of 10 mM acetic acid solution diluted with an equal amount of Freund’s adjuvant. Serial dorsal skin biopsies were performed at 7, 15, and 30 days and stained with H&E, Masson’s trichrome and Picrosírius for morphological and morphometric analyses.

RESULTS:

Immunized rabbits presented a significant increase in collagen in skin collected seven days after the first immunization (p=0.05).

CONCLUSION:

The results from this experimental model may be very important to a better understanding of the pathogenic mechanisms involved in the beginning of human SSc. Therapeutic protocols to avoid early remodeling of the skin may lead to promising treatments for SSc in the future.     

Roles of Periostin in Symptom Manifestation and Airway Remodeling in a Murine Model of Allergic Rhinitis

Purpose

Periostin was originally identified as a secreted factor during screening of a mouse osteoblastic library. In a recent study, periostin was found to directly regulate eosinophil accumulation in allergic mucosal inflammation. Chronic eosinophilic inflammation is related to the development of remodeling. The present study examined the expression of periostin and evaluated its role in the inflammatory process and remodeling associated with allergic rhinitis.

Methods

A murine model of allergic rhinitis was established in periostin knockout mice. We analyzed the expression of periostin, manifestation of nasal symptoms, eosinophilic inflammation, and subepithelial fibrosis as well as the expression of MMP-2, TIMP-1, and type 1 collagen in nasal tissue.

Results

Periostin was mainly distributed in the subepithelial tissue of the nasal mucosa. The subepithelial tissue was thinner in the knockout group than in the control group. No differences in the expression of MMP-2 or TIMP-1 were found in the knockout group. However, after a month of allergen challenge, type I collagen in the nasal tissue was lower in the knockout group than in the control group. The number of eosinophils and the symptom score were also lower in the knockout group.

Conclusions

Periostin is expressed in nasal tissues of murine models of allergic rhinitis. Periostin deficiency may affect the remodeling of nasal tissue with reduced subepithelial fibrosis, and lead to less eosinophilic inflammation.