OBJECTIVETo investigate hydroxyproline tolerance of Chrysanthemum morifolium plantlets included " Boju", "Huaiju", "Chuju", "Gongju" and "Hangju",and provide references basis for excellent cultivar and breeding of Ch. morifolium.

METHODPlantlets in vitro from five kinds of Ch. morifolium were inoculated on medium added with different concentrations of hydroxyproline. Free proline in leaves from plantlets was determined, then the damage index and survival rate were compared.

RESULT AND CONCLUSIONThe results showed that hydroxyproline tolerance of " Boju" and "Huaiju" were superior, the survival rates and free proline of them were higher, but the damage index was inferior. The hydroxyproline tolerance of "Hangju" was the worst, and the survival rate was minimum. The survival rate of "Chuju" and "Gongju" was between "Boju" and " Hangju", and the hydroxyproline tolerance of them was also medium.

Chrysanthemum ; chemistry ; classification ; metabolism ; Culture Techniques ; Hydroxyproline ; analysis ; metabolism

PURPOSE: This study was designed to measure time-dependent changes in muscle excursion and collagen content after tenotomy, and to analyze the correlation between muscle excursion and collagen content in a rabbit model. MATERIALS AND METHODS: Twenty-four rabbits underwent tenotomy of the second extensor digitorum longus (EDL) muscles on the right legs and were randomly assigned to three groups based on the period of time after tenotomy (2, 4, and 6 weeks). The second EDL muscles on left legs were used as controls. At each time after tenotomy, passive muscle excursion and collagen content, determined by hydroxyproline content, were measured bilaterally, and the ratio of each value to the normal one was used. RESULTS: The mean ratio of muscle excursion after tenotomy to the value of the control decreased in a time-dependent fashion: 92.5% at 2 weeks, 78.6% at 4 weeks, and 55.1% at 6 weeks. The mean ratio of hydroxyproline content in muscle to the value of the control increased in a time-dependent fashion: 119.5% at 2 weeks, 157.3% at 4 weeks, and 166.6% at 6 weeks. There was a significant negative correlation between the ratio of hydroxyproline content in muscle after tenotomy to the control values and the ratio of muscle excursion after tenotomy to the control values (r=-0.602, p=0.002). CONCLUSION: The decrease in muscle excursion seems to correlate with the increase in collagen content in the muscle in a time-dependent fashion following tenotomy.
Animals ; Collagen/*metabolism ; Hydroxyproline/metabolism ; Muscle, Skeletal/*metabolism ; Rabbits ; Tendon Injuries/*metabolism ; Tendons ; Tenotomy ; Time Factors

BACKGROUNDGastrojejunostomy is one of the most frequently used procedures for general surgeons. The creation of anastomosis between various parts of the gastrointestinal tract is a basic technical component and major task in the daily practice of almost all gastrointestinal procedures. This research evaluated a new procedure of making gastrointestinal anastomosis with stent.

METHODSTwenty experimental mini-pigs were randomized into two groups. In stent anastomosis group (SA), the anastomoses were constructed with a poly-levolactic acid stent. In hand-sewn group (HA), the anastomoses were performed with a single-layer continuous suture. Abdominal X-ray with intraluminal contrast was performed on the 10th postoperative day. Five pigs of each group were sacrificed on the postoperative days 3 and 14 to determine anastomotic bursting pressure in situ, hydroxyproline concentration, and histopathological evaluation of the anastomotic sites.

RESULTSThere was no intraoperative morbidity or mortality. The median time needed for the sutured anastomosis was (21.7 ± 2.3) minutes and for the stent anastomosis was (11.9 ± 1.9) minutes (P < 0.001). Abdominal X-ray with intraluminal contrast demonstrated normal gas distribution and showed no evidence of leakage or obstruction. Macroscopic appearance at the longitudinal opening of anastomosis was always good in both groups. The median anastomotic bursting pressure was (18.2 ± 1.6) kPa in SA group on postoperative day 3, compared with (11.7 ± 3.2) kPa in HA group (P = 0.003). The anastomotic bursting pressure on day 14 was not significantly different between SA group ((27.1 ± 2.6) kPa) and HA group ((28.3 ± 1.7) kPa) (P = 0.388). The hydroxyproline concentrations were not significantly different.

CONCLUSIONSThe stent anastomosis was not considered to be more difficult than a sutured anastomosis. This method is proved to be safe and feasible compared with the traditional hand-sewn method in the porcine model. The method increases early anastomotic strength in this study.

Anastomosis, Surgical ; methods ; Animals ; Female ; Gastric Bypass ; methods ; Hydroxyproline ; metabolism ; Male ; Stents ; Swine

Animals ; Disease Models, Animal ; Female ; Fibrosis ; metabolism ; pathology ; Hydroxyproline ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Thrombospondin 1 ; pharmacology

Animals ; Brain ; pathology ; Hydroxyproline ; metabolism ; Kidney ; pathology ; Liver ; pathology ; Lung ; metabolism ; pathology ; Male ; Paraquat ; metabolism ; poisoning ; Rats ; Rats, Wistar

A porcine heart valve was irradiated by Ultraviolet (UV) rays (10 W, 254 nm) for 2, 4, 8 and 24 hours at 4 degrees C to cross-link the structural collagen matrix. The degree of cross-linking was evaluated by assaying the released amount of hydroxyproline (Hyp) from the matrix, and comparing it with the positive controls of valves treated by glutaraldehyde (GA) solution (0.625 wt%) and the negative controls of non-treated fresh valves. The undigested weight ratio of the specimens increased by increasing the UV irradiation time. The undigested weight of the leaflets, tunica interna and tunica externa of the fresh, GA-treated and UV-irradiated specimens after collagenase digestion was compared. As UV irradiation increased, the amount of released hydroxyproline was gradually decreased until 8 hours of irradiation, after which the released hydroxyproline-reduction occurred slightly until 24 hours of irradiation time in this system. A total 47.68% of the hydroxyproline in the valve was cross-linked by UV irradiation after 24 hours, while 73.74% of the hydroxyproline in the positive control was crossed-linked. Light microscopic observation revealed that the typical crimp pattern of collagen fibers decreased and was rearranged into a dense flattened pattern as the UV irradiation induced interfibrilar cross-linking. GA-treated valves demonstrated a denser matrix pattern than the UV-irradiated specimens. Cross-linked collagenous tissue prepared by UV irradiation would be useful for improving durability and reducing the disadvantages related to using a chemical cross-linking agent.
Animal ; Aortic Valve/radiation effects* ; Aortic Valve/metabolism* ; Collagen/radiation effects* ; Collagen/chemistry* ; Hydroxyproline/metabolism ; Swine ; Ultraviolet Rays*

OBJECTIVETo study the proliferation and apoptosis in carbon tetrachloride induced rat liver fibrosis.

METHODSWistar rats were injected subcutaneously with 40% CCl4-olive oil twice weekly for 12 weeks. Liver tissues were obtained at the end of 4, 8, 12 and 16 weeks for histological examination, hydroxyproline (Hyp) assay and proteomic analysis. After two dimension electrophoresis (2-DE), the silver stained gels were analyzed with PDQUEST 2-DE. More than 30 differentially expressed proteins were identified by MALDI-TOF-MS.

RESULTSThe degree of collagen deposition and hydroxyproline content of the fibrotic livers increased continuously during the 12 weeks of CCl4 administration, peaked at the end of week 12 (P < 0.05) and declined significantly at week 16 (P < 0.05). Significant differences were observed in two parameters at each time point between the control and the model group. Meanwhile, dramatic change of hepatic proteome in the model group rats was also seen. Differentially expressed proteins identified by MALDI-TOF-MS were categorized as proliferation-related proteins/enzymes (proliferating cell nuclear antigen p120, p40 and cyclin F ubiquitin-conjugating enzyme 7 UBC7), and apoptosis-related proteins, mainly caspase-12 which was absent in the control rats.

CONCLUSIONProliferation and apoptosis related proteins are expressed dynamically in different stages of rat liver fibrosis induced by CCl4.

Animals ; Apoptosis ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Caspase 12 ; metabolism ; Cell Proliferation ; Hydroxyproline ; metabolism ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Proteins ; metabolism ; Proteome ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo explore the quantity of mast cells and the role of protease activated receptor-2 (PAR-2) in experimental rat liver fibrosis.

METHODSRats were sacrificed at 0, 2, 4, 8, and 12 weeks after subcutaneous injection of CCl(4). Mast cells were displayed by toluidine blue stain. The content of liver hydroxyproline was measured by the method of base hydrolyzate. The mRNA expression and the protein expression of PAR-2 in livers were detected by RT-PCR and immunohistochemistry at each time point.

RESULTSIn normal rat livers there were a few mast cells (2.5+/-1.0) distributed along the hepatic portal areas. In the cirrhosis model group the number of mast cells in the livers increased degree by degree (2 weeks vs 4 weeks vs 8 weeks, 9.1+/-0.5 vs 15.7+/-3.0 vs 32.0+/-3.3; P less than 0.05), and they were distributed densely around the hepatic portal areas and the central veins. The content of liver hydroxyproline increased progressively from 0 to 12 weeks. In normal livers PAR-2 mRNA was hardly detected, at 2 weeks there was some expression of PAR-2 mRNA (PAR-2/beta-actin 0.15+/-0.01, P less than 0.05), at 4 weeks its expression increased (PAR-2/beta-actin 0.35+/-0.02, P less than 0.05) and it maintained a higher level (PAR-2/beta-actin 0.80+/-0.02, P less than 0.05) since then. The changing trend of the protein expression of PAR-2 was the same as that of PAR-2 mRNA expression.

CONCLUSIONSPAR-2 mRNA expression and the protein expression of PAR-2 were consistent with the increase of the mast cells, and the content of liver hydroxyproline may play an important role in mediating liver fibrosis.

Animals ; Hydroxyproline ; metabolism ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Male ; Mast Cells ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, PAR-2 ; metabolism

OBJECTIVETo explore the stimulation of human hepatic stellate cells by Cytochrome P4502E1-mediated oxidative stress.

METHODSHepG2-line was transfected with human CYP2E1 plasmid (HepG2/CYP2E1) and empty plasmid (HepG2/PCI) respectively. The CYP2E1 expression was evaluated with RT-PCR and Western blot. MDA was measured in culture medium of HepG2 cell lines. LX2 was co-incubated with HepG2/CYP2E1, HepG2/PCI and HepG2 respectively. The level of hydroxyproline in culture medium was examined in 48 hours and the cells were lysated and total RNA and protein were extracted. COL-1 and MMP2 mRNA levels were detected by RT-PCR and analyzed semi-quantitatively. PICP proteins were measured by ELISA. Zymography was performed to investigate MMP2 enzymatic activities.

RESULTS(1) MDA from the HepG2 which (HepG2/CYP2E1)express human CYP2E1 (6.51+/-0.25) was significantly higher than that from the HepG2 which do not (HepG2/PCI) express human CYP2E1 (3.07+/-0.29) and HepG2 alone (2.57+/-0.29). (F=22.66, all P<0.01). (2) After co-incubated for 48 hours,the level of hydroxyproline in culture medium (35.24+/-3.52) excreted from CYP2E1/LX2 could significantly increase (F=58.89, P is less than 0.01). PICP protein (540.01+/-11.38) excreted from CYP2E1/LX2 was significantly increased (F=124.97, P<0.01). Zymography showed MMP2 gene expression and enzymatic activities of MMP2 had no difference among the groups (F=0.29, P>0.05) (F=0.33, P>0.05).

CONCLUSIONSCYP2E1 derived oxidative stress mediated stimulation of collagen I synthesis by hepatic stellate cells. Hydroxyproline excreted by LX2 was increased by CYP2E1. COL-1mRNA had no difference among the groups (F=0.73, P>0.05).

Collagen Type I ; biosynthesis ; Cytochrome P-450 CYP2E1 ; metabolism ; Hep G2 Cells ; Hepatic Stellate Cells ; metabolism ; Humans ; Hydroxyproline ; secretion ; Liver Cirrhosis ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Oxidative Stress

OBJECTIVETo observe the therapeutic effect and mechanism of penehyclidine hydrochloride on paraquat-induced acute lung injury.

METHODS80 healthy adult male Wistar rats were randomly assigned into control groups (10 rats), 100 mg/kg PQ group (10 rats), 100 mg/kg PQ plus 33 µg/kg penehyclidine hydrochloride treatment group (30 rats), 100 mg/kg PQ plus 66 µg/kg penehyclidine hydrochloride treatment group (30 rats). The two treatment groups were executed respectively at 36 h, 72 h and 7 d. Lung tissues were used to assess histopathological change by HE staining. The level of MMP-2, caveolin-1 and HYP were detected in the lung homogenate. The serum and BALF contents of ET were measured.

RESULTSPathology inspection confirmed that the model of acute rat pulmonary injury were duplicated successfully. The level of MMP-2, HYP in lung tissues and the serum and BALF ET contents in PQ group were (1.77 ± 0.40) µg/g, (2.91 ± 0.79) µg/g, (505.23 ± 124.69) µg/ml, (640.38 ± 136.60) µg/ml. The level of those was higher than that in control group [(0.95 ± 0.66) µg/g, (1.48 ± 0.69) µg/g, (95.48 ± 46.01) µg/ml, (200.40 ± 88.39) µg/ml, P < 0.05]; The above-mentioned index in two treatment groups was lower than that in PQ group (P < 0.05). The caveolin-1 content [(1.77 ± 0.82) µg/g] in PQ group was lower than that in control group [(5.39 ± 1.68) µg/g, P < 0.05]. The level of caveolin-1 in two treatment groups was higher than that in PQ group (P < 0.05).

CONCLUSIONPenehyclidine hydrochloride can decrease the level of MMP-2, HYP in lung tissues and the ET in serum and BALF, increase that of caveolin-1 and lessen the damage induced by paraquat.

Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Caveolin 1 ; metabolism ; Endothelins ; metabolism ; Hydroxyproline ; metabolism ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Paraquat ; toxicity ; Quinuclidines ; therapeutic use ; Rats ; Rats, Wistar