Animals ; Collagen Type I ; metabolism ; Collagen Type II ; metabolism ; Collagen Type III ; metabolism ; Female ; Kinetin ; pharmacology ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism

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

OBJECTIVEThe aim of this study was to investigate the expression of collagen in the process of masseter muscle reattachment to the cortical and cancellous bones of mandible.

METHODSA total of nine adult goats were used in the study. One was the control. The other eight were treated with bilateral detachment of the masseter muscles. The biopsies of bone and muscle were taken at 2, 4, 8 and 12 weeks after the operation. The characteristics of the healing muscle-bone interfaces were examined using immunohistochemical techniques.

RESULTSImmunohistochemical analysis illustrated that the locations of collagen type I, II and III were different during the healing process, but similar in the cortical and cancellous bones.

CONCLUSIONThis study demonstrates that the distribution of the three types of collagens at the muscle-bone interfaces is associated with time, but not related with their locations.

Animals ; Collagen ; biosynthesis ; genetics ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type II ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Female ; Goats ; Male ; Mandible ; metabolism ; pathology ; surgery ; Masseter Muscle ; metabolism ; pathology ; surgery ; Wound Healing ; physiology

The present study was aimed to investigate the role and mechanism of glutaminolysis of cardiac fibroblasts (CFs) in hypertension-induced myocardial fibrosis. C57BL/6J mice were administered with a chronic infusion of angiotensin II (Ang II, 1.6 mg/kg per d) with a micro-osmotic pump to induce myocardial fibrosis. Masson staining was used to evaluate myocardial fibrosis. The mice were intraperitoneally injected with BPTES (12.5 mg/kg), a glutaminase 1 (GLS1)-specific inhibitor, to inhibit glutaminolysis simultaneously. Immunohistochemistry and Western blot were used to detect protein expression levels of GLS1, Collagen I and Collagen III in cardiac tissue. Neonatal Sprague-Dawley (SD) rat CFs were treated with 4 mmol/L glutamine (Gln) or BPTES (5 μmol/L) with or without Ang II (0.4 μmol/L) stimulation. The CFs were also treated with 2 mmol/L α-ketoglutarate (α-KG) under the stimulation of Ang II and BPTES. Wound healing test and CCK-8 were used to detect CFs migration and proliferation respectively. RT-qPCR and Western blot were used to detect mRNA and protein expression levels of GLS1, Collagen I and Collagen III. The results showed that blood pressure, heart weight and myocardial fibrosis were increased in Ang II-treated mice, and GLS1 expression in cardiac tissue was also significantly up-regulated. Gln significantly promoted the proliferation, migration, mRNA and protein expression of GLS1, Collagen I and Collagen III in the CFs with or without Ang II stimulation, whereas BPTES significantly decreased the above indices in the CFs. α-KG supplementation reversed the inhibitory effect of BPTES on the CFs under Ang II stimulation. Furthermore, in vivo intraperitoneal injection of BPTES alleviated cardiac fibrosis of Ang II-treated mice. In conclusion, glutaminolysis plays an important role in the process of cardiac fibrosis induced by Ang II. Targeted inhibition of glutaminolysis may be a new strategy for the treatment of myocardial fibrosis.
Rats ; Mice ; Animals ; Rats, Sprague-Dawley ; Angiotensin II/pharmacology* ; Fibroblasts ; Mice, Inbred C57BL ; Fibrosis ; Collagen/pharmacology* ; Collagen Type I/metabolism* ; RNA, Messenger/metabolism* ; Myocardium/pathology*

OBJECTIVETo look into the character of the expression of collagen type II and transforming growth factor beta1 (TGF-beta1), basic fibroblast growth factor (bFGF) in the apical articular process cartilages of adolescent idiopathic scoliosis (AIS) and congenital scoliosis (CS) patients.

METHODSThe articular processes of 22 AIS and 18 CS were collected. The techniques of HE staining, immunohistochemistry and in situ hybridization were adopted in this research. By comparing the apical processes with the end processes, the convex processes with the concave processes, the AIS processes with CS processes, the pathological changes of the articular process cartilages of these patients and the distribution of collagen type II and TGF-beta1, bFGF in them were studied. The images of immunohistochemistry and in situ hybridization were input into the image analysis system and were analyzed semi-quantitatively. The SAS software (8.01) was adopted, and P < 0.05 was defined as the significant level.

RESULTSThe expression of collagen type II and TGF-beta1, bFGF in AIS was similar to CS: the concave sides of apexes were higher than the convex sides. The comparisons had statistical significance. There was no statistical significance between upper and lower end vertebrae in convex and concave sides, between convex and concave sides in upper and lower end vertebrae. The apical vertebrae were significantly higher than the ipsilateral sides of upper or lower end vertebrae for collagen type II. There was no statistical difference of the expression at the concave, convex, upper, lower end vertebrae between AIS and CS.

CONCLUSIONSThe cartilages of the apical processes show some signs of regression and hypoplasia in scoliosis. The concave side is more severe than the convex side. Increase of collagen type II and TGF-beta1, bFGF in the concave sides of apical processes in scoliosis may be the results of reconstruction of extracellular matrix and the compensation reactions which are caused by abnormal biomechanical forces such as compressive stresses. Compressive stress on the concave sides has more influences on the expression of collagen type II than tensile stress on the convex sides.

Adolescent ; Cartilage, Articular ; metabolism ; pathology ; Child ; Collagen Type II ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Scoliosis ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo observe the influence of Pilose antler polypeptide on the glycosaminoglycan and type II collagen in the articular cartilage in experimental knee osteoarthritis.

METHODSTotally 64 New Zealand white rabbits of 6 months old were randomly divided into 2 groups:normal group (n = 8) and model group (n = 56). Model group was surgically induced into osteoarthritis model by method of Hulth. After successful modeling, the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group and control group, 24 rabbits in each group. Pilose antler polypeptide-treatment group received 0.5 ml intra-articular injection of Pilose antler polypeptide dilution liquid once in per 2 days for 30 days, while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples were collected respectively. The content of glycosaminoglycan in articular cartilage was observed by toluidine blue staining and the expression of type II collagen in cartilage matrix was detected by immunohistochemical staining.

RESULTSAlong with the prolonging of time, the content of glycosaminoglycan and type II collagen in cartilage matrix of the Pilose antler polypeptide-treatment group and control group decreased gradually. On days 7, 15 and 30 after intervention, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group were (312.06 +/- 14.12), (273.31 +/- 12.42) and (248.34 +/- 10.41), which had statistically significant differences. Integrated optical density of the type II collagen positive area in cartilage matrix of the control group were (253.47 +/- 15.53), (215.67 +/- 9.72) and (160.01 +/- 13.23), which had statistically significant differences. At the same period, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group was higher than that of control group, which had statistically significant difference.

CONCLUSIONPilose antler polypeptide can inhibit reduction of the glycosaminoglycan and type II collagen in cartilage matrix and delay the degeneration of articular cartilage.

Animals ; Antlers ; chemistry ; metabolism ; Collagen Type II ; metabolism ; Disease Models, Animal ; Female ; Glycosaminoglycans ; metabolism ; Humans ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Peptides ; metabolism ; pharmacology ; Rabbits

BACKGROUNDQuantitative T2 mapping has been a widely used method for the evaluation of pathological cartilage properties, and the histological assessment system of osteoarthritis in the rabbit has been published recently. The aim of the study was to investigate the effectiveness of quantitative T2 mapping evaluation for articular cartilage lesions of a rabbit model of anterior cruciate ligament transection (ACLT) osteoarthritis.

METHODSTwenty New Zealand White (NZW) rabbits were divided into ACLT surgical group and sham operated group equally. The anterior cruciate ligaments of the rabbits in ACLT group were transected, while the joints were closed intactly in sham operated group. Magnetic resonance (MR) examinations were performed on 3.0T MR unit at week 0, week 6, and week 12. T2 values were computed on GE ADW4.3 workstation. All rabbits were killed at week 13, and left knees were stained with Haematoxylin and Eosin. Semiquantitative histological grading was obtained according to the osteoarthritis cartilage histopathology assessment system. Computerized image analysis was performed to quantitate the immunostained collagen type II.

RESULTSThe average MR T2 value of whole left knee cartilage in ACLT surgical group ((29.05±12.01) ms) was significantly higher than that in sham operated group ((24.52±7.97) ms) (P=0.024) at week 6. The average T2 value increased to (32.18±12.79) ms in ACLT group at week 12, but remained near the baseline level ((27.66±8.08) ms) in the sham operated group (P=0.03). The cartilage lesion level of left knee in ACLT group was significantly increased at week 6 (P=0.005) and week 12 (P<0.001). T2 values had positive correlation with histological grading scores, but inverse correlation with optical densities (OD) of type II collagen.

CONCLUSIONThis study demonstrated the reliability and practicability of quantitative T2 mapping for the cartilage injury of rabbit ACLT osteoarthritis model.

Animals ; Anterior Cruciate Ligament ; metabolism ; physiopathology ; Cartilage, Articular ; metabolism ; physiopathology ; Collagen Type II ; metabolism ; Magnetic Resonance Imaging ; Male ; Osteoarthritis ; metabolism ; physiopathology ; Rabbits

BACKGROUND: N-containing bisphosphonates (BPs), such as pamidronate and risedronate, can inhibit osteoclastic function and reduce osteoclast number by inducing apoptotic cell death in osteoclasts. The aim of this study is to demonstrate the effect of pamidronate, second generation nitrogen-containing BPs and to elucidate matrix metallo-proteinases (MMPs) mRNA expression under serum starvation and/or tumor necrosis factor alpha (TNF-α) stimulation on metabolism of intervertebral disc (IVD) cells in vitro. METHODS: Firstly, to test the effect of pamidronate on IVD cells in vitro, various concentrations (10⁻¹², 10⁻¹⁰, 10⁻⁸, and 10⁻⁶ M) of pamidronate were administered to IVD cells. Then DNA and proteoglycan synthesis were measured and messenger RNA (mRNA) expressions of type I collagen, type II collagen, and aggrecan were analyzed. Secondly, to elucidate the expression of MMPs mRNA in human IVD cells under the lower serum status, IVD cells were cultivated in full serum or 1% serum. Thirdly, to elucidate the expression of MMPs mRNA in IVD cells under the stimulation of 1% serum and TNF-α (10 ng/mL) In this study, IVD cells were cultivated in three dimensional alginate bead. RESULTS: Under the lower serum culture, IVD cells in alginate beads showed upregulation of MMP 2, 3, 9, 13 mRNA. The cells in lower serum and TNF-α also demonstrated upregulation of MMP-2, 3, 9, and 13 mRNA. The cells with various doses of pamidronate and lower serum and TNF-α were reveled partial down-regulation of MMPs. CONCLUSIONS: Pamidronate, N-containing second generation BPs, was safe in metabolism of IVD in vitro maintaining chondrogenic phenotype and matrix synthesis, and down-regulated TNF-α induced MMPs expression.
Aggrecans ; Cell Death ; Collagen ; Collagen Type I ; Collagen Type II ; Diphosphonates ; DNA ; Down-Regulation ; Humans* ; In Vitro Techniques ; Intervertebral Disc* ; Matrix Metalloproteinases* ; Metabolism ; Osteoclasts ; Phenotype ; Proteoglycans ; Risedronate Sodium ; RNA, Messenger ; Starvation ; Tumor Necrosis Factor-alpha* ; Up-Regulation

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Hepatocytes ; cytology ; metabolism ; Tetrazoles ; pharmacology ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

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