[Role of CTGF and PI3K/Akt signaling pathway in paraquat-induced mesenchymal changes in alveolar epithelial cells].

Objective: To investigate the role of connective tissue growth factor (CTGF) and PI3K/Akt signaling pathways in paraquat (PQ) -induced alterations in alveolar epithelial cell mesenchymalization (EMT) . Methods: In February 2023, RLE-6TN cells were divided into 2 groups, which were set as uncontaminated group and contaminated group (200 µmol/L PQ), and cellular EMT alteration, CTGF and PI3K/Akt signaling pathway related molecules expression were detected by cell scratch assay, qRT-PCR and western-blot assay. Using shRNA interference technology to specifically inhibit the expression of CTGF, RLE-6TN cells were divided into four groups: control group, PQ group (200 µmol/L PQ), interference group (transfected with a plasmid with shRNA-CTGF+200 µmol/L PQ), and null-loaded group (transfected with a plasmid with scramble- CTGF+200 µmol/L PQ), qRT-PCR and western blot were used to examine the alteration of the cellular EMT and the expression of molecules related to the activity of PI3K/Akt pathway. The PI3K/Akt signaling pathway was blocked by the PI3K inhibitor LY294002, and the expression of EMT-related molecules in cells of the control group, PQ group (200 µmol/L PQ), and inhibitor group (200 µmol/L PQ+20 µmol/L LY294002) was examined by qRT-PCR and western blot.The t-test was used to compare the differences between the two groups, while the analysis of variance (ANOVA) was applied to compare the differences among multiple groups. For further pairwise comparisons, the Bonferroni method was adopted. Results: The results of cell scratch test showed that compared with the uncontaminated group, RLE-6TN cells in the contaminated group had faster migration rate, lower mRNA and protein expression levels of E-Cadherin, and higher mRNA and protein expression levels of α-SMA, CTGF, PI3K and Akt, with statistical significance (P<0.05). After specific inhibition of CTGF expression, the mRNA and protein expression of CTGF, PI3K, Akt, and α-SMA in the cells of the interference group were significantly lower than that of the PQ group and the null-loaded group (P<0.05/6), whereas that of E-Cadherin was higher than that of the PQ group and the null-loaded group (P<0.05/6). Specifically blocking the PI3K/Akt signaling pathway, the mRNA and protein expression of PI3K, Akt and α-SMA in the cells of the inhibitor group was decreased compared with that of the PQ group (P<0.05/3), while the expression of E-Cadherin was elevated compared with that of the PQ group (P<0.05/3) . Conclusion: CTGF may promote PQ-induced alveolar epithelial cell EMT through activation of the PI3K/Akt signaling pathway. Inhibition of CTGF expression or blockade of PI3K/Akt signaling pathway activity can alleviate the extent of PQ-induced alveolar epithelial cell EMT.

[Experience of mesenteric sector fixation in mesenteric torsion treatment].

Objective: The preliminary results was reported regarding the treatment of mesenteric torsion by mesenteric fixation in the last decade, especially preventing recurrence of mesenteric torsion by mesenteric fan-shaped fixation. Methods: We selected 12 patients who received emergency operation in Chongqing Hospital of the First Affiliated Hospital of Guangzhou University of Chinese Medicine from December 2010 to March 2022. All of them were made a definite diagnose of mesenteric torsion by the preoperative CT scan or exploratory laparotomy. The recurrence of mesenteric torsion will be prevented by taking the operation of mesenteric fan-shaped fixation. This technique is suitable for the patient who is suffering total mesenteric torsion, but enteric necrosis is excluded affirmatively. The operation is consists of the following progress: (1) Exploratory laparotomy to check for necrosis of the bowel and for lesions other than torsion. (2) Mesenteric torsion derotation.(3) Mesenteric linear fixation; the right posterior lower border of the small mesentery (terminal ileal mesentery) is intermittently sutured to the posterior peritoneum of the right lower quadrant to increase the width of the base of the small mesentery. (4) Mesenteric fan-shaped fixation, which is fan-shaped to the lower left and fixed in the posterior peritoneum, shortening the length of the mesentery and further increasing the width of the mesentery and posterior peritoneal fixation. Results: A total of 12 patients with mesenteric torsion were treated by operation for 15 times in all. Among them, 3 cases received resection of most small bowel were performed without recurrence; 3 patients received only derotation for a total of 4 times, 2 cases recurred, 1 of them recurred twice; 4 cases underwent derotation and mesenteric linear fixation,and 1 case recurred. Four patients with derotation and mesenteric fan-shaped fixation recovered well without recurrence. Conclusion: Mesenteric fan-shaped fixation may be an effective operative type to reduce or avoid postoperative recurrence of mesenteric torsion.

Effect of Ageratina adenophora on hepatic and pulmonic pathological lesions in horses.

The effect of Ageratina adenophora on pathological characteristics of the liver and lungs as well as serum biochemical parameters in horses were investigated. Ten horses without ingestion history of Ageratina adenophora were classified into the control group, and 10 poisoned but survived horses with 3 months ingestion history were set as the case group. Results showed that serum AST, ALT, ALP, magnesium and phosphorus were elevated significantly, while creatinine was decreased remarkably. Hematoxylin and eosin staining of liver tissues showed diffuse swelling or destruction of hepatocytes, narrowing or atrophy of the hepatic sinusoids, and little lymphocytic infiltration; lung tissues presented destroyed alveoli and inflammatory cell infiltration.

MiR-181b regulates atherosclerotic inflammation and vascular endothelial function through Notch1 signaling pathway.

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-181b on the inflammation and vascular endothelial function in atherosclerosis (AS), and its specific molecular regulatory mechanism. MATERIALS AND METHODS: 44 apolipoprotein E (ApoE)-/- 7 weeks old male rats were randomly divided into normal diet group (NC group) and AS model group (high-fat diet feeding). Rat aorta was dissected and the serum sample was collected in both groups. The serum levels of inflammatory factors in both groups were detected via enzyme-linked immunosorbent assay (ELISA). The mRNA levels of miR-18b and Notch1 were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Moreover, aortic endothelial cells were extracted from AS rats. The mir-18b binding target gene was analyzed via bioinformatics and further verified by the Luciferase reporter gene assay. The protein expressions of miR-18b and Notch1 in endothelial cells transfected with miR-181b mimic or inhibitor were detected. Influence of miR-181b on vascular endothelial indexes was also detected. RESULTS: Compared with those in the NC group, the serum levels of interleukin-6 (IL-6), IL-10, IL-13 and tumor necrosis factor-α (TNF-α) in the AS group significantly increased (p<0.05). The mRNA level of miR-18b in AS plaques was significantly lower than that in NC arterial tissues. Conversely, Notch1 level in AS plaques was markedly higher than that in the NC arterial tissues (p<0.05), with the mean difference of 2.12 and 2.82 folds (p<0.05). According to the Pearson correlation analysis, there was a significant negative correlation between mRNA levels of miR-181b and Notch1 in AS tissues (r=-0.65, p=0.014). The bioinformatics analysis showed that there were complementary binding sites between miR-181b and Notch1. The Luciferase reporter gene assay confirmed the presence of direct binding sites between miR-181b and Notch1. Western blotting revealed that the overexpression of miR-181b downregulated Notch1 and hs-CRP, but upregulated BNP (p<0.05). Opposite trends were obtained by miR-181b knockdown (p<0.05). CONCLUSIONS: The decline in the miR-181b expression may be an important factor in AS plaque formation and vascular endothelial injury by regulating Notch1.

Effects of MiR-21 on the proliferation and migration of vascular smooth muscle cells in rats with atherosclerosis via the Akt/ERK signaling pathway.

OBJECTIVE: To explore the effects and mechanism of action of micro ribonucleic acid (miR)-21 on the proliferation and migration of vascular smooth muscle (VSM) in atherosclerosis (AS). MATERIALS AND METHODS: The rats were fed with a high-fat diet, and the oil red staining was adopted to compare AS between Sprague Dawley (SD) rats and miR-21 knockdown rats. At the in-vitro level, primary rat VSM cells (VSMCs) were selected and divided into miR-NC blank control group [miR-normal control (NC) group] and miR-21 overexpression group (miR-21 group) for relevant experimental detection. Wound healing assay and transwell assay were used to detect the effects of miR-21 on the proliferation and migration of VSMCs. Westernn blotting was applied to examine the changes in the levels of Cyclin D, a cell cycle-related protein, and the key factors of the Akt/ERK signaling pathway, such as phosphorylated-Akt (p-AKT), AKT, p-ERK1/2, and ERK1/2. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell activity assay kit was applied to determine the effects of miR-21 on the proliferation of VSMCs through regulating the Akt/ERK signaling pathway after the ERK signaling pathway inhibitor PD98059 and AKT inhibitor MK-2206 were given. RESULTS: Compared with that in miR-NC group, the level of AS in miR-21 knockdown rats were decreased significantly (p < 0.05). In the cell-level experiment, the overexpression of miR-21 promoted abnormal proliferation of VSMCs and activated the Akt/ERK signaling pathway (p < 0.05). MTT assay results revealed that inhibiting the Akt/ERK pathway could reverse the effects of miR-21 promoting proliferation and migration. CONCLUSIONS: MiR-21 promotes the proliferation and migration of VSMCs by activating the Akt/ERK pathway and aggravates AS. Knocking down miR-21 or inhibiting the Akt/ERK pathway can suppress the activation of VSMCs.

IL-10 targets Th1/Th2 balance in vascular dementia.

OBJECTIVE: The pathogenesis of vascular dementia (VD) is not fully elucidated. Th1/Th2 balance may change in VD, leading to numerous inflammatory cytokines secretion. Interleukin-10 (IL-10) is an immune suppressor, while its function in VD and correlation with Th1/Th2 balance are still unclear. MATERIALS AND METHODS: The healthy male rats were randomly divided into three groups, including sham group, model group, and IL-10 group. Th1 and Th2 cytokines IL-2, IL-4, IL-6, and tumor necrosis factor-α (TNF-α) expressions in the serum were tested by enzyme-linked immunosorbent assay (ELISA). IL-10 expression in brain tissue and peripheral blood was detected by Real-Time PCR and ELISA. The correlation relationship between IL-10 and T helper cells 1/2 (Th1/Th2) cytokines was analyzed. Hippocampus cell apoptosis was determined by caspase 3 activity kit. Nuclear transcription factor 2 κB (NF-κB) expression was evaluated by Western blot. RESULTS: IL-10 levels were decreased, caspase 3 activity was enhanced, NF-κB expression was declined, IL-2 and TNF-α secretion were up-regulated, while IL-4 and IL-6 secretion were reduced in hippocampus tissue and peripheral blood from VD model rat compared to sham group (p<0.05). IL-10 significantly attenuated caspase 3 activity, up-regulated NF-κB expression, reduced IL-2 and TNF-α secretion, and enhanced IL-4 and IL-6 secretion (p<0.05). IL-10 was negatively correlated with Th1 cytokines and positively correlated with Th2 cytokines (p<0.05). CONCLUSIONS: IL-10 expression declined in VD and participated in regulating Th1/Th2 balance. IL-10 participated in VD incidence and development through regulating cell apoptosis and NF-κB expression.

[Establishment of multiple predictor models of severe acute pancreatitis in elderly patients].

Objective: To investigate the prognostic factors related to the severity of acute pancreatitis and to establish the multiple predictor models of severe acute pancreatitis(SAP) in elderly patients. Methods: Clinical data of 146 consecutive elderly patients who met the inclusion criteria between January 2014 and May 2017 at Department of Pancreatic and Biliary Surgery, the First Affiliated Hospital of Harbin Medical University were retrospectively collected and analyzed, wherein 88 cases were mild acute pancreatitis, 29 cases were moderately severe acute pancreatitis and 29 cases were SAP. The patients data were subjected to univariate analysis and multiple classified Logistic regression analysis for independent prognostic factors of the severity of acute pancreatitis in elderly patients. Unweighted predictive score(unwScore) and weighted predictive score(wScore)for SAP in elderly patients were established according which the receiver-operating characteristic(ROC) curves of independent prognostic factors and predictor models were produced. The cutoff values of independeut prognostic factors and predictor models were determined. The area under the curve, the sensitivity, the specificity, the positive predictive value and the negative predictive value to verify the predictive efficiency of the independent prognostic factors and predictor models were calculated. Results: Procalcitonin(PCT)(Z=10.564, P=0.000), blood urea nitrogen(BUN)(Z=22.231, P=0.003), serum creatinine(Scr)(Z=14.151, P=0.030), serum calcium(Z=34.979, P=0.032) and pleural effusion(χ(2)=28.463, P=0.015) were independent prognostic factors of the severity of acute pancreatitis by univariate analysis and multiple classified Logistic regression analysis in elderly patients. Respectively, the area under the curve of PCT, BUN, Scr, serum calcium and pleural effusion were 0.908, 0.737, 0.701, 0.753, 0.712, the sensitivity were 0.828, 0.621, 0.552, 0.690, 0.517, the specificity were 0.915, 0.786, 0.846, 0.966, 0.906, the positive predictive value were 70.6%, 41.9%, 47.1%, 83.3%, 57.7%, the negative predictive value were 95.5%, 89.3%, 88.4%, 92.6%, 88.3%. Respectively, the area under the curve of unwScore and wScore were 0.915 and 0.953, the sensitivity were 0.759 and 0.931, the specificity were 0.889 and 0.915, the positive predictive value were 62.9% and 73.0%, the negative predictive value were 93.7% and 98.2%. Conclusions: PCT, BUN, Scr, serum calcium and pleural effusion were independent prognostic factors of the severity of acute pancreatitis in elderly patients. The multiple predictor models of SAP in elderly patients have a good predictive efficiency, which may provide valuable clinical reference for prediction and treatment.

Peroxiredoxin1 promotes cell proliferation, migration and invasion of colorectal cancer via p38MAPK signaling.

OBJECTIVE: Peroxiredoxin1 (PRDX1), a class of thiol peroxidases, is a multifunctional protein. We aimed at analyzing the effect of PRDX1 on proliferation, apoptosis, migration and invasion of colorectal cancer and to investigate the potential mechanism. MATERIALS AND METHODS: Western blot and PCR were used to validate the silencing efficiency in SW480 cell by transfection of PRDX1-siRNA. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) test. Flow cytometry Annexin V/PI double staining was used to analyze cell apoptosis. Transwell and scratch test were used to detect the migration and invasion of cells. Signal pathway protein expression was analyzed by Western blot. RESULTS: The expression of PRDX1 in SW480 cells could be reduced by siRNA effectively. The cell proliferation, migration and invasion were reduced significantly compared with control group after down-regulation of PRDX1 (p<0.05), while the cell apoptosis was enhanced significantly (p<0.05). The ratio of phospho-p38 mitogen-activated protein kinases (p-p38) /p38 mitogen-activated protein kinases (p38) was down-regulated after the down-regulation of PRDX1 (p<0.05). The ratio of phospho-c-Jun N-terminal protein kinase (p-JNK)/c-Jun N-terminal protein kinase (JNK) and phospho-extracellular regulated protein kinases (p-ERK)/extracellular regulated protein kinases (ERK) showed changes with no significant difference (p>0.05). CONCLUSIONS: Down-regulation of PRDX1 in colorectal cancer SW480 cells could inhibit the cell proliferation, migration, invasion, and induce cell apoptosis. This is very likely to be achieved by activating the p38MAPK-signaling pathway.

Prediction of morbidity following cytoreductive surgery for metastatic gastrointestinal stromal tumour in patients on tyrosine kinase inhibitor therapy.

BACKGROUND: Although cytoreductive surgery has been shown to be beneficial in carefully selected patients with metastatic gastrointestinal stromal tumours (GISTs) treated with tyrosine kinase inhibitors (TKIs), factors predictive of postoperative morbidity have not been investigated previously. METHODS: A surgical complexity score for GIST metastasectomy (GM-SCS) composed of patient-related and surgical factors was assigned retrospectively to patients with metastatic GIST treated with TKI therapy and surgery at two institutions between 2002 and 2014. The ability of clinicopathological factors and GM-SCS to predict postoperative morbidity was assessed by means of a multivariable logistic regression model. Postoperative complications were categorized using the Clavien-Dindo classification. RESULTS: Some 400 operations on 323 patients with metastatic GIST on TKIs were included. Complications were observed following 110 operations (27·5 per cent) including 70 major complications (grade III-V) (17·5 per cent of 400 operations). Patients were divided into low (5 points or less; 100 patients, 25·0 per cent), intermediate (6-9 points; 191, 47·8 per cent) and high (at least 10 points; 109, 27·3 per cent) complexity scoring groups based on the GM-SCS. An intermediate (odds ratio (OR) 2·88; P = 0·008) and high (OR 5·40; P < 0·001) GM-SCS were independent predictors of overall complications, whereas only a high GM-SCS was independently predictive of a major complication (OR 3·65; P = 0·018). Metastatic mitotic index was also an independent predictor of overall complications (OR 2·55; P = 0·047). GM-SCS did not predict progression-free or overall survival. CONCLUSION: A gastrointestinal stromal tumour metastastectomy surgical complexity score can predict morbidity, which may help in preoperative risk stratification and optimal treatment planning.

Protection against ischemia-reperfusion injury in aged liver donor by the induction of exogenous human telomerase reverse transcriptase gene.

BACKGROUND: Liver ischemia-reperfusion (I/R) injury is of great importance in primary graft dysfunction after transplantation, and could be more severe in transplantation using aged donor livers. In order to alleviate the I/R injury in aged donor livers, we transferred exogenous human telomerase reverse transcriptase (hTERT) gene into aged rat's livers before liver transplantation. After transplantation, the effect of the gene for aged rats on cell apoptosis caused by I/R injury was evaluated. METHODS: The experiment was divided into 2 parts: comparative experiment between aged rats and adult rats, and exogenous induction experiment of aged rats. In the first part, Wistar rats were divided into 2 groups; group I was composed of adult rats (5 months) and group II was composed of aged rats (16-18 months). After successful transplantation, chronic oxidative stress and lipid peroxidation-related indicators (contents of vitamin C and vitamin E; activities of superoxide dismutase, catalase, and methane decarboxylic aldehyde) and alanine aminotransferase activity were examined. In the second part, additional aged rats were divided into 3 groups: group A included the donors pretreated with exogenous hTERT gene; group B included the donors pretreated with adenovirus vector; and group C was composed of the donors pretreated with physiological saline. Various indicators were detected to analyze the effect of the gene on I/R injury of the aged rats. RESULTS: The lower vitamin C, vitamin E, SOD, and CAT contents in the aged group than those in the adult group (P < .05), and the higher MDA and ALT contents in the aged group than those in the adult group (P < .05) were observed. The apoptotic index and ALT levels in the hTERT gene-pretreated group were significantly lower than those in the adenovirus vector group and the physiological saline group (P < .05). Meanwhile, mild histological injury and increased telomerase activity were also observed in the hTERT gene-pretreated group. CONCLUSION: Compared with the adult rats, I/R injury in the aged liver donor is more severe. The induction of exogenous hTERT gene offers protection against I/R injury in the aged liver.

Application of tissue-engineered cartilage with BMP-7 gene to repair knee joint cartilage injury in rabbits.

Injured articular cartilage has a poor capacity for spontaneous healing. So far, satisfactory solution to this subsistent problem has not been found, but transgenic therapy may be a promising way. This study aims to evaluate the effectiveness of a tissue-engineered cartilage that was transfected with morphogenetic protein 7 (BMP 7) in repairing the cartilaginous defects of rabbit knee joints. Chondrocytes were transfected with BMP-7 gene (5 x 10(6) cells/ml), inoculated into the collagen-fibrin gel scaffolds, and cultured for 14 days. Then, the scaffolds were implanted onto the created defects (5.0 mm in diameter) in rabbits' knee joints. After 12 weeks, the rabbits were sacrificed and histological sections were evaluated using modified O'Driscoll cartilage scores; In situ hybridization and immunohistochemistry were performed to detect the expression of BMP-7 mRNA and BMP-7 at the implanted site while the content of DNA and GAG was determined as well. A better quality of repairs was observed at the 12th week after implantation when compared to the control group using histological analyses. The content of DNA and specific secretion of GAG in the treatment group is statistically significant different compared with the control group. Gene therapy may be a promising treatment method, but the novel therapy approach needs further studies with respect to a longer follow-up period.

Antioxidant defenses of mycorrhizal fungus infection against SO(2)-induced oxidative stress in Avena nuda seedlings.

Colonization of arbuscular mycorrhizal fungi Glomus mosseae increased Avena nuda seedling tolerance to SO(2) exposure, as indicated by elevated total plant biomass and ameliorative photosynthetic rate, when compared to the non-mycorrhizal plants. This is associated with an improved antioxidant capacity as shown by enhanced superoxide dismutase and catalase activity, increased ascorbic acid and glutathione content, and reduced malondialdehyde and hydrogen peroxide level in the mycorrhizal plants relative to the non-mycorrhizal plants under SO(2) exposure. The mycorrhizal fungi colonization had no effect on the stomatal conductance. To our knowledge, this is the first finding of this sort.

Prostate cancer detection: comparison of T2-weighted imaging, diffusion-weighted imaging, proton magnetic resonance spectroscopic imaging, and the three techniques combined.

BACKGROUND: Magnetic resonance (MR) imaging has been established as the best imaging modality for the detection, localization, and staging of prostate cancer on account of its high resolution of soft tissue and the possibilities of multiplanar and multiparameter scanning. PURPOSE: To evaluate T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), MR spectroscopy (MRS), and the combination of these three MR techniques in the diagnosis of prostate cancer, as correlated to histopathologic findings. MATERIAL AND METHODS: MR imaging, including T2WI, DWI, and MRS, was performed at 1.5T with a body coil combined with a spine coil in 42 cases. Diagnosis was confirmed by histopathology through systematic transrectal prostate biopsy. Apparent diffusion coefficient (ADC) maps were obtained with two b values (0 and 1000 s/mm(2)). The metabolic maps of 3D-MRS imaging were analyzed for the ratio of (Cho+Cre)/Cit in each sextant. All cases were evaluated by T2WI, DWI, and MRS, and then by the three methods combined. The statistical indicators and the receiver operating characteristic (ROC) curve analysis of each method were compared to the results of histopathology obtained by transrectal prostate biopsy. RESULTS: 15 of 42 cases were confirmed to be cancerous, and 27 of 42 cases were noncancerous. All the 252 sextants were confirmed by biopsies, including 201 benign sextants and 51 malignant sextants. The sensitivity and the specificity for the detection of prostate cancer were 88.2% and 67.2% for T2WI, as the cutoff was 3; 82.4% and 81.6% for DWI, as the cutoff was 4; 84.3% and 98.0% for MRS, as the cutoff was 5; and 96.1% and 96.5% for the combined T2WI+DWI+MRS, as the cutoff was 4. In the ROC analysis, the correlative areas under the ROC curves (Az) were 0.848+/-0.030, 0.860+/-0.033, and 0.961+/-0.016 for T2WI, DWI, and MRS, respectively, and 0.978+/-0.009 for the combination of T2WI+DWI+MRS. CONCLUSION: The accuracy of the detection of prostate cancer is increased through a combination of the three techniques. Moreover, MRS demonstrated higher accuracy compared with T2WI or DWI.

The role of adrenomedullin and its receptor system in cardiovascular calcification of rat induced by Vitamin D(3) plus nicotine.

Adrenomedullin (ADM) is a potent vasodilatory peptide which regulates blood pressure, cell growth and bone formation. Our work was aimed to explore the production of ADM, changes and pathophysiological significance of ADM mRNA and ADM receptor components--calcitonin receptor like receptor (CRLR) and receptor activity modifying proteins (RAMPs) mRNA in calcified myocardium and aorta of rats induced by Vitamin D3 plus nicotine. Contents of ADM in plasma, myocardium and aorta were measured by radioimmunoassay (RIA). The amount of ADM, CRLR and RAMPs mRNA was determined by semi-quantitative RT-PCR. The calcium content and alkaline phosphatase activity in myocardium and aorta of rats were measured. The results showed that the contents of calcium in calcified myocardium and aorta were increased by 3.5- and 6-fold (all P < 0.01), respectively, and alkaline phosphatases activity in calcified myocardium and aorta were increased by 66.5 and 82.7% (all P < 0.01 ), respectively, compared with control. Contents of ADM in plasma, myocardium and aorta were increased by 58% (P < 0.01), 14.3% (P < 0.01) and 27.8% P < 0.05). Furthermore, it was found that the amount of ADM, CRLR and RAMP2 mRNA in calcified myocardium was elevated by 90.6, 157.5 and 119.6% (all P < 0.01), RAMP3 mRNA was decreased by 14.1% (P < 0.01), respectively, compared with control. The amount of ADM, CRLR, RAMP2 and RAMP3 mRNA in calcified aorta was elevated by 37.7% (P < 0.01), 41.4% (P < 0.01), 60.1% (P < 0.05) and 13% P < 0.01), respectively, compared with control. The elevated level of CRLR and RAMP2 mRNA were in positive correlation with that of ADM mRNA (r = 0.992 and 0.882, respectively, P < 0.01) in calcified myocardium. The elevated level of CRLR and RAMP3 mRNA were also in positive correlation with that of ADM mRNA (r = 0.727, P < 0.05 and 0.816, P < 0.01, respectively) in calcified aorta. These results demonstrated that calcified myocardium and aorta generated an increased amount of ADM, up-regulated gene expressions of ADM, CRLR and RAMP2 mRNA. While the alteration of RAMP3 mRNA in calcified myocardium and aorta was different. These suggested that ADM and its receptor system might involve in the regulation of calcification in heart and aorta.

[Formation of the allogeneic tissue engineered cartilage using injectable biomaterial].

OBJECTIVE: To study the feasibility of the formation of allogeneic tissue-engineered cartilage of certain shape in immunocompetent animal using the injectable biomaterial. METHODS: Fresh newborn rabbits' articular cartilages were obtained under sterile condition (< 6 hours after death) and incubated in the sterile 0.3% type II collagenase solution. After digestion of 8 to 12 hours, the solution was filtered through a 150 micron nylon mesh and centrifuged, then the chondrocytes were washed twice with phosphate buffered saline (PBS) and mixed with the biomaterial to create a final cell density of 5 x 10(7)/ml. The cell-biomaterial admixture was injected into rabbits subcutaneously 0.3 ml each point while we drew the needle back in order to form the neocartilage in the shape of cudgel, and the control groups were injected with only the biomaterial or the suspension of chondrocytes with the density of 5 x 10(7)/ml. After 4, 6, 8 and 12 weeks, the neocartilages were harvested to analyze. RESULTS: The new nodes could be touched subcutaneously after 2 weeks. In the sections of the samples harvested after 4 weeks, it was found that the matrix secreted and the collagen formed. After 6 weeks and later than that, the neocartilages were mature and the biomaterial was almost completely degraded. The cudgel-shaped samples of neocartilage could be formed by injection. In the experiment group, there was no obvious immune rejection response. On the contrary, there were no neocartilage formed in the control group. CONCLUSION: The injectable biomaterial is a relatively ideal biomaterial for tissue engineering, and it is feasible to form allogeneic tissue engineered cartilage of certain shape by injection in an immunocompetent animal.

[Experimental study on repair of cricoid cartilage defect with cartilage grafts in rabbits].

OBJECTIVE: To investigate the effect of free cartilage fragment in repair of cricoid cartilage defect. METHOD: One-third of cricoid cartilage was cut and repaired with free thyroid cartilage fragment and pedicled cartilage in rabbits. The pathological changes of the cartilages were observed. RESULT: 1. The free thyroid cartilage fragments were absorbed in two months, and were replaced by scars. 2. The pedicled cartilage fragments were not absorbed and healed with the cricoid cartilage. 3. No evidence of growth impairment of surgery-induced cricoid cartilage was observed. CONCLUSION: In rabbits, both free cartilage fragments and pedicled cartilage fragments could repair the defect of cricoid cartilage effectively.

Effects of MK-447 on thrombin-induced aggregation, secretion of ATP, and [Ca2+]i mobilization in rabbit platelets.

AIM: To study the effects of MK-447 on aggregation release reaction and intracellular calcium mobilization by thrombin. METHODS: Aggregation and release reaction were assessed by light transmission and ATP content in rabbit citrate platelet-rich plasma (PRP), and cytosolic-free calcium was measured by fluorescence and imaging. RESULTS: MK-447 (2-aminomethyl-4-t-butyl-6-iodophenol hydrochloride) induced a decrease in light transmission (DLT), so called platelet shape change, without detectable aggregation and secretion of ATP, and increased intracellular calcium concentration ([Ca2+]i) slightly in washed single platelet loaded with Fura 2, the peak value being about 160 nmol.L-1. These effects were not inhibited by egtazic acid 3 mmol.L-1 or indometacin 3 mumol.L-1. The pretreatment of PRP with MK-447 700 mumol.L-1 reduced the DLT by thrombin, potentiated and enhanced thrombin-induced aggregation and secretion of ATP in a concentration-dependent manner. Thrombin-induced [Ca2+]i mobilization (peak value: 369 +/- 45 nmol.L-1) was further enhanced by the administration of MK-447 at 2 min before the addition of thrombin, and the peak value reached 623 +/- 121 nmol.L-1 (P < 0.01). CONCLUSION: MK-447-induced platelet shape change was involved in intracellular calcium release in this preparation. MK-447 enhanced thrombin-induced aggregation and release reaction and these effects of MK-447 on aggregation and release reaction by thrombin might result from the synergistic effect of intracellular calcium mobilization.

[Clinical observation on treatment of myelodysplastic syndromes with yisuikang].

20 cases of myelodysplastic syndrome (MDS) patients were treated with Yisuikang (YSK) including MDS-RA 17 cases, MDS-RAEB 2 cases, MDS-RAEB-T 1 case. 14 patients were treated with YSK only, 6 cases were treated with both YSK and Western medicine. The result showed that 4 cases were remitted, 5 cases markedly improved, 6 cases improved and 5 cases ineffective, the total effective rate was 75.0%. It was found that YSK was effective in the treatment of MDS-RA or Qi-Blood Deficiency and Yu-Re Syndrome in TCM, while any other types were ineffective. They also found that types of TCM might be correlated with MDS.

Altered regulation of IL-6 production with normal aging. Possible linkage to the age-associated decline in dehydroepiandrosterone and its sulfated derivative.

Normal aging in humans has been recently shown to be accompanied by reduced control over production of the multifunctional cytokine IL-6. This cytokine was reported to be quantitatively elevated in most serum samples obtained from "normal" elderly humans. In the present investigation, we report that IL-6 levels are elevated in serum samples obtained from aged mice, and its spontaneous production could also be easily detected in culture supernatants of unstimulated lymphoid cells obtained from aged, but not mature, adult donors. Spontaneous production of IL-6 was consistently observed in culture supernatants of lymphoid cells from both the spleen and mesenteric lymph nodes from aged donors, but was absent from supernatants derived from their peripheral lymph nodes. In aged mice, the reduced regulation of IL-6 production could be effectively prevented and/or reversed by supplementing aging animals with dehydroepiandrosterone sulfate, a steroid hormone whose endogenous production is known to decline with advancing age in all species tested. It was also established that serum obtained from old dehydroepiandrosterone sulfate-treated mice contained lower (normal) levels of serum amyloid P substance (an acute phase reactant), reduced levels of serum Ig (all classes and isotypes) and lower titers of tissue-specific autoantibodies than untreated aged controls. Therefore, a number of well described, age-related conditions, some of which could be contributing to the pathologic phenotype of old age, may actually represent secondary effects to this age-associated change in IL-6 production.

Administration of dehydroepiandrosterone to burned mice preserves normal immunologic competence.

Burned individuals display a reduced ability to elicit cellular and humoral immune responses and a depression in the vitro production of certain T-cell lymphokines. Treatment of burned mice with 100 micrograms of dehydroepiandrosterone within 1 hour after injury resulted in preserving a completely normal capacity to produce T-cell-derived lymphokines and to generate cellular immune responses. In addition, dehydroepiandrosterone-treated thermally injured mice demonstrated an above-normal ability to resist an induced infection with the intracellular pathogen, Listeria monocytogenes. Dehydroepiandrosterone-treated animals also did not exhibit the sustained plasma levels of interleukin 6 that normally accompany thermal injury and infection. Because of its antiglucocorticoid effects and positive immunoregulatory influences, we believe dehydroepiandrosterone to be a beneficial form of therapy for thermally injured individuals.