Collagen Type I ; analysis ; Collagen Type III ; analysis ; Fluorescent Antibody Technique, Indirect ; Humans ; Liver ; chemistry ; Microscopy, Confocal

OBJECTIVETo investigate the correlation and anatomic association of benign hyperplastic nodules in the peripheral zone (PZ) with those in the transition zone (TZ) of the prostate, and to compare the histological components of the two kinds of nodules.

METHODSWe obtained benign hyperplastic nodules specimens from the PZ and TZ by autopsy, measured the distance between the outer surface of the nodules and the inner gland, observed the integrity of the surgical envelope of the prostate, and determined the histological components of the two kinds of nodules by HE staining, immunohistochemistry and automatic quantitative image analysis.

RESULTSThe surgical envelope of the prostate was integrated and the distance between the nodules of the PZ and the outer surface of the inner gland was about 2.5 to 5 mm ([3.9 +/- 0.8] mm), with no signs of anatomic connection in between. The stromata and epithelia in the nodules accounted for (69.32 +/- 8.35)% and (16.08 +/- 5.36)% in the PZ and (74.58 +/- 8.95)% and (15.82 +/- 6.41)% in the TZ.

CONCLUSIONBenign hyperplastic nodules may originate from the PZ of the prostate and not correlate with the inner gland hyperplasia in the TZ, but with no statistical difference between the histological components of the two kinds of nodules.

Aged ; Aged, 80 and over ; Autopsy ; Collagen Type I ; analysis ; Collagen Type II ; analysis ; Collagen Type III ; analysis ; Collagen Type IV ; analysis ; Fibronectins ; analysis ; Humans ; Hyperplasia ; Immunohistochemistry ; Laminin ; analysis ; Male ; Prostate ; chemistry ; pathology ; Prostatic Hyperplasia ; metabolism ; pathology

A method for quantitation of collagen was established by detecting marker peptide with high performance liquid chromatography-mass spectrometry (HPLC-MS). Theoretical marker peptides were selected by sequence comparison. Bovine collagen type I was digested with trypsin. Marker peptides typical for collagen type I were identified with HPLC-MS. The relationship between the abundance of marker peptides and collagen concentration was established. The results show that GEAGPSGPAGPTGAR and the other 5 peptides showed high resolution during chromatographic separation and high signal intensity during MS analysis. Peptide signal intensity and collagen concentration showed a good linear relationship in the range from 0.1 to 3 mg/mL. Bovine tendon and collagen sponge were used as actual samples and collagen contents were determined as 90.2% and 93.4% respectively. Quantitation of marker peptides of collagen was a feasible method to identify and quantify collagens in medical device research and development.
Animals ; Cattle ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; Mass Spectrometry ; Peptides ; analysis

OBJECTIVETo investigate the effect of a Chinese herbal prescription on collagen I in rat's femur under simulated weightlessness.

METHODSThirty Wistar rats were randomly divided into 3 groups: blank control group (10 rats), tail suspension group (TS, 10 rats), TS with Chinese medicine group (10 rats). Rats in TS with Chinese medicine group took a Chinese herbal prescription (contains Radix Rehmanniae Praeparata, Radix Achyranthis Bidentatae, Radix Astragali, Radix Angelicae Sinensis, Concha Ostreae prepared by acetic acid)by oral administration. After 1 week adaption and 3 weeks tail suspending, rat's left femur was colleced, and collagen I in femur neck was detected by immunohistochemical method.

RESULTSCounts and integral optical density (IOD) of collagen I coloration decreased significantly in TS group (P < 0.001), but no significant change in TS with Chinese medicine group (P > 0.05), as compared with control group.

CONCLUSIONGeneration of collagen I become weaken under simulated weightlessness, while the Chinese herbal prescription is effective to prevent the change, thus biochemistry environment of bone calcium deposition may be improved by this Chinese herbal prescription under simulated weightlessness.

Animals ; Collagen Type I ; analysis ; Drugs, Chinese Herbal ; pharmacology ; Femur ; chemistry ; drug effects ; Immunohistochemistry ; Male ; Rats ; Rats, Wistar ; Weightlessness Simulation

OBJECTIVETo study the effects of excessive fluoride on type I collagen in rat developmental dentine.

METHODSEighty SD rats, 5 days old, were divided into experimental and control groups, 40 in each group. The experimental group received subcutaneous injection of 0.2% NaF every 4 days (the dose was 2 mg NaF per kg body wt). The same volume of 0.9% NaCl was used in the control. Twenty rats in each group were killed 4 days after the second and the seventh injection respectively. The expression of type I collagen was assayed with immunohistochemical technique.

RESULTS4 out of 20 rats after two injections showed abnormal distribution of type I collagen (dense stain of collagen in the odontoblast, aggregation of collagen in the dentine and disordered arrangement of collagen in the predentine; All 20 rats after seven injections showed abnormal distribution of type I collagen.

CONCLUSIONExcessive fluoride may affect the metabolism of type I collagen in rat developmental dentine.

Animals ; Animals, Newborn ; Collagen Type I ; analysis ; Dentin ; chemistry ; drug effects ; growth & development ; Fluorides ; toxicity ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley

Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to 125.0+/-4.0% (mean+/-SD), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to 222.3+/-33.8%. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to 149.2+/-2.8%, and increased the degree of mineralization, a marker of the late process of differentiation, to 122.9+/-3.9%. Rutin alone also increased the activity of ALP (to 154.4+/-12.2%), the expression of collagen type I (to 126.6+/-6.2%), and the degree of mineralization (to 112.3+/-5.0%). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.
Alkaline Phosphatase ; Bone Resorption ; Cell Proliferation ; Cell Survival ; Collagen Type I ; Humans ; Osteoblasts ; Osteogenesis ; Osteoporosis ; Rutin* ; Spectrum Analysis

BACKGROUNDBone morphogenetic protein (BMP)-2, alkaline phosphatase (ALP), and collagen type I are known to play a critical role in the process of bone remodeling. However, the relationship between mechanical strain and the expression of BMP-2, ALP, and COL-I in osteoblasts was still unknown. The purpose of this study was to investigate the effects of different magnitudes of mechanical strain on osteoblast morphology and on the expression of BMP-2, ALP, and COL-I.

METHODSOsteoblast-like cells were flexed at four deformation rates (0, 6%, 12%, and 18% elongation). The expression of BMP-2 mRNA, ALP, and COL-I in osteoblast-like cells were determined by real-time quantitative reverse transcription polymerase chain reaction, respectively. The results were subjected to analysis of variance (ANOVA) using SPSS 13.0 statistical software.

RESULTSThe cells changed to fusiform and grew in the direction of the applied strain after the mechanical strain was loaded. Expression level of the BMP-2, ALP, and COL-I increased magnitude-dependently with mechanical loading in the experimental groups, and the 12% elongation group had the highest expression (P < 0.05).

CONCLUSIONMechanical strain can induce morphological change and a magnitude-dependent increase in the expression of BMP-2, ALP, and COL-I mRNA in osteoblast-like cells, which might influence bone remodeling in orthodontic treatment.

Alkaline Phosphatase ; metabolism ; Analysis of Variance ; Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Cell Line ; Collagen ; metabolism ; Collagen Type I ; metabolism ; Mice ; Osteoblasts ; cytology ; metabolism

OBJECTIVEThe etiology of developmental dislocation of the hip (DDH) remains uncertain, but some research has shown that this disorder is closely related to hip joint laxity. This study examined the expression of collagens type I and III mRNA and protein in the hip capsule of children with DDH in order to investigate the roles of collagens type I and III in hip joint laxity.

METHODSNine children with DDH and nine age and gender-matched normal children (control group) were enrolled. Semiquantitative RT-PCR method was used to detect mRNA expression of COL1a1 and COL3a1 in the hip capsule. Western-Blot method was used to detect protein expression of COL1a1 and COL3a1 in the hip capsule. The quantitative analysis of the COL1a1 and COL3a1 was performed by professional image software and the results were analyzed with standard statistical methods.

RESULTSmRNA and protein expression of COL1a1 in the DDH group was significantly lower than that in the control group (P<0.01). Compared with the control group, COL1a3 mRNA expression in the DDH group decreased significantly (P<0.01), but COL1a3 protein expression was not significantly different.

CONCLUSIONSThe decreased collagen I mRNA and protein expression in the hip capsule might contribute to hip joint laxity in children with DDH. Collagen type III may not be associated with hip joint laxity in DDH.

Blotting, Western ; Child ; Child Development ; Child, Preschool ; Collagen Type I ; analysis ; genetics ; Collagen Type III ; analysis ; genetics ; Female ; Hip Dislocation ; metabolism ; Humans ; Infant ; Male ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo analyze the constituent proteins in donkey hide, the key ingredient for Ejiao, an important traditional Chinese medicine for the blood-related conditions, in hope to eventually decipher the biochemical mechanism behind Ejiao's prominent medicinal efficacy.

METHODTwo methods were employed to extract proteins in donkey skin. One used TriPure isolation reagent to extract the total proteins in donkey skin. Another used 1% sodium dodecyl sulfate (SDS) to heat the sample at 100 degrees C overnight. And then sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and capillary HPLC were used to analyze the component of proteins.

RESULTThere are not only collagen alpha1 (I) and collagen alpha2 (I), but also serum albumin in donkey skin. The content is over 25% in total proteins with the method of TriPure isolation reagent. The content of donkey serum albumin is up to 20% with the method of 1% SDS heating. And two bands, molecular weight are nearly 200 kDa,were found on 7.5% SDS-PAGE. Extracted these proteins to analyze with capillary HPLC, they were found to be the complex products of collagen and serum albumin of donkey.

CONCLUSIONDonkey serum albumin is a main protein component in the hide, which is a clue to expose is the effect of Ejiao on blood.

Animals ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; chemistry ; metabolism ; Collagen Type II ; analysis ; chemistry ; metabolism ; Drug Interactions ; Electrophoresis, Polyacrylamide Gel ; Equidae ; Molecular Weight ; Protein Binding ; Serum Albumin ; analysis ; chemistry ; metabolism ; Skin ; chemistry

OBJECTIVETo investigate the dynamic changes of expression of matrix metalloproteinases-9 in myocardium of mice with viral myocarditis (VMC) and its significance in the pathogenesis of viral myocarditis.

METHODSVMC model was prepared by an injection of CVB3 in BALB/C mice. The mice receiving an injection of culture solution without virus were used as the control group. Cardiac tissues were obtained 7, 14, 21 and 28 days after injection and made into paraffin sections. Myocardial histopathologic changes were observed by hematoxylin-eosin staining and Masson staining. The expression of MMP-9, type I collagen and type III collagen in cardiac tissues were quantified by SABC immunohistochemical method.

RESULTSThe expression of MMP-9 in the VMC model group was observed on the 7th day, reached a peak on the 14th day, and was significantly higher than that in the control group at all time points (P<0.05). Compared with the control group, the expression of type I collagen in the VMC model group was up-regulated on the 21st day and reached a peak on the 28th day (P<0.05). The expression of type III collagen in the VMC model group was significantly higher than that in the control group on the 28th day (P<0.05). The expression of MMP-9 was positively correlated with myocardial histopathologic scores (r=0.832, P<0.05) and negatively correlated with type I collagen expression (r=-0.791, P<0.05).

CONCLUSIONSMMP-9 is over-expressed at the early stage in VMC mice, and participates in the pathological process of VMC through mediating the degradation metabolism of type I collagen. It may be an important factor that leads to myocardial collagen remodeling and myocardial fibrosis.

Animals ; Collagen Type I ; analysis ; Collagen Type III ; analysis ; Coxsackievirus Infections ; enzymology ; Enterovirus B, Human ; Immunohistochemistry ; Male ; Matrix Metalloproteinase 9 ; analysis ; Mice ; Mice, Inbred BALB C ; Myocarditis ; enzymology ; pathology ; Myocardium ; enzymology ; pathology