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Objective To establish a fast and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation.Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations` distribution,each tube of cell fraction was then partially transfered to the 96 plate well,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy.Results Three types of spermatogenic cells can be identified quickly and accurately by it`s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection.Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation is successfully developed.
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<p><b>OBJECTIVE</b>To assess the effect of laparoscopic colectomy on the exfoliated cancer cells in peritoneal cavity, recurrence and metastasis of patients with colonic carcinoma.</p><p><b>METHODS</b>One hundred and fifty-nine patients with colonic cancer proven by colonoscopy and pathology were divided into two groups based on patient's preference: laparoscopic group (n=74) and open group (n=85). The positive rate of exfoliated cancer cells in peritoneal cavity was compared by cytological detection before and after cancer resection. Recurrence, metastasis rate and 3-year survival were compared between the two groups.</p><p><b>RESULTS</b>The positive rates of exfoliated cancer cells in peritoneal cavity were 12.2% (9/74) in the laparoscopic group and 15.3% (13/85) in the open group before cancer resection without significant difference (P=0.718); 20.3% (15/74) and 30.6% (26/85) after cancer resection without significant difference (P=0.138). The follow-up ranged from 4 to 45 months. The 3-year local recurrence rates were 13.6% (8/59) and 8.8% (6/68) (P=0.455), the 3-year distal metastasis rates were 11.9% (7/59) and 17.6% (12/68) (P=0.416) and the 3-year survival rates were 79.7% and 80.0% (P=0.998), and the differences were not statistically significant.</p><p><b>CONCLUSION</b>The laparoscopic operation does not increase the recurrence and metastasis rate and results in similar survival in patients with colonic cancer as compared to open procedure.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Colonic Neoplasms , Diagnosis , General Surgery , Laparoscopy , Neoplasm Metastasis , Neoplasm Recurrence, Local , Peritoneal Cavity , Pathology , Prognosis , Survival RateABSTRACT
We aimed to examine the effect of pioglitazone on transdifferentiation of preosteoblasts from rat bone marrow mesenchymal stem cells (BMSCs) into adipocytes and investigate its effect on bone metabolism. BMSCs were harvested from the femurs and tibias of a rat, then separated, purified, proliferated for 3 generations and differentiated into preosteoblasts for 5 days and 14 days respectively in the presence of osteogenic medium. Thereafter, the preosteoblasts were cultured for 21 days in the presence of adipogenic medium with and without pioglitazone (1 μg/mL). Partially-differentiated osteoblasts were identified by mineralized nodules with Alizarin red S staining. Transdifferentiated adipocytes were identified by Oil Red O staining. Reverse transcription PCR (RT-PCR) was performed to assay the expression levels of osteogenic markers Runx2 and ALP, and an adipogenic marker PPARγ. Those cells cultured for 5 days did not show mineralized nodules as detected by staining of Alizarin red S, while those cultured for 14 days showed dispersed mineralized centers in the form of brown spots, although without obvious red mineralized nodules. After adipogenic transdifferentiation for 21 days, adipose-drops were found in cells of 5CG and 5EG earlier than those of 14CG and 14EG, and the former showed much more adipocytes separately as detected by Oil Red O staining. Whatever the time was 5 days or 14 days of BMSCs osteogenic differentiation, the cells cultured with pioglitazone showed much more adipocytes than those without pioglitazone. Our experiment showed that the less time it took for BMSCs osteogenic differentiation, a stronger ability remained for BMSCs to transdifferentiate into adipocytes. The mRNA expression levels of Runx2 and ALP were decreased by 1.79 and 1.90 times respectively in 5EG (P< 0.05) as compared with 5CG, and that of PPARγ was increased by 1.31 times in 5EG (P<0.05) as compared with 5CG. The mRNA expression levels of Runx2 and ALP were decreased by 1.45 and 1.54 times respectively in 14EG (P<0.05) as compared with 14CG, and that of PPARγ was increased by 1.39 times in 14EG (P<0.05) as compared with 14CG. It was concluded that pioglitazone stimulated the transdifferentiation of BMSCs into adipocytes. These observations provided a potential mechanism of imbalance in thiazolidinedione induced bone metabolism.
ABSTRACT
We aimed to examine the effect of pioglitazone on transdifferentiation of preosteoblasts from rat bone marrow mesenchymal stem cells (BMSCs) into adipocytes and investigate its effect on bone metabolism. BMSCs were harvested from the femurs and tibias of a rat, then separated, purified, proliferated for 3 generations and differentiated into preosteoblasts for 5 days and 14 days respectively in the presence of osteogenic medium. Thereafter, the preosteoblasts were cultured for 21 days in the presence of adipogenic medium with and without pioglitazone (1 μg/mL). Partially-differentiated osteoblasts were identified by mineralized nodules with Alizarin red S staining. Transdifferentiated adipocytes were identified by Oil Red O staining. Reverse transcription PCR (RT-PCR) was performed to assay the expression levels of osteogenic markers Runx2 and ALP, and an adipogenic marker PPARγ. Those cells cultured for 5 days did not show mineralized nodules as detected by staining of Alizarin red S, while those cultured for 14 days showed dispersed mineralized centers in the form of brown spots, although without obvious red mineralized nodules. After adipogenic transdifferentiation for 21 days, adipose-drops were found in cells of 5CG and 5EG earlier than those of 14CG and 14EG, and the former showed much more adipocytes separately as detected by Oil Red O staining. Whatever the time was 5 days or 14 days of BMSCs osteogenic differentiation, the cells cultured with pioglitazone showed much more adipocytes than those without pioglitazone. Our experiment showed that the less time it took for BMSCs osteogenic differentiation, a stronger ability remained for BMSCs to transdifferentiate into adipocytes. The mRNA expression levels of Runx2 and ALP were decreased by 1.79 and 1.90 times respectively in 5EG (P< 0.05) as compared with 5CG, and that of PPARγ was increased by 1.31 times in 5EG (P<0.05) as compared with 5CG. The mRNA expression levels of Runx2 and ALP were decreased by 1.45 and 1.54 times respectively in 14EG (P<0.05) as compared with 14CG, and that of PPARγ was increased by 1.39 times in 14EG (P<0.05) as compared with 14CG. It was concluded that pioglitazone stimulated the transdifferentiation of BMSCs into adipocytes. These observations provided a potential mechanism of imbalance in thiazolidinedione induced bone metabolism.
Subject(s)
Animals , Female , Male , Rats , Adipocytes , Cell Transdifferentiation , Mesenchymal Stem Cells , Osteoblasts , Osteogenesis , Rats, Sprague-Dawley , Thiazolidinediones , PharmacologyABSTRACT
Objective To investigate the relationship between EphA7 protein and carcinogenesis and development of pancreatic cancer by detecting the expression of EphA7 protein in pancreatic cancer tissues. Methods The expression of EphA7 in 10 cases of normal pancreatic tissue, 51 cases of pancreatic cancer and its adjacent tissues were detected with immunohistechemieal methods and the relationship between EphA7 and pathologic features of pancreatic cancer were analyzed. Results The rate of expression of EphA7 protein in normal pancreatic tissue was 10% (1/10), 47.1% (24/51) in adjacent pancreatic cancer tissues, 94.1% (48/51) in pancreatic cancer tissues. There were significant difference among the three groups (P <0.05). There was no significant correlation between the expression of EphA7 protein and the age, sex, tumor location, tumor size in patients with pancreatic cancer (P > 0.05). However there was significant correlation between the expression of EphA7 protein and the degree of differentiation, clinical staging, lymph node metastasis, or distant metastasis (P < 0.05). Conclusions The abnormally high expression of EphA7 may be relevant with the occurrence and development of the pancreatic cancer.
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The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen activation or NF-kappaB activation in rats was studied in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-kappaB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10(-3) mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P 0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-kappaB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.
Subject(s)
Carbachol/pharmacology , Cholinergic Agonists/pharmacology , NF-kappa B/metabolism , Pancreas/metabolism , Pancreas/pathology , Rats, Wistar , Receptor, Muscarinic M3/agonists , Trypsinogen/metabolismABSTRACT
The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulationinduced pancreatic acinar cellular injury and trypsinogen activation or NF-κB activation in rats was studied in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-κB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10-3 mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P ＜0.01) following the treatment with a high concentration of carbachol (10-3 mol/L) in vitro. The addition of 10-2 mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10-3 mol/L) in vitro (P＞0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-κB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro.