ObjectiveTo explore the effect of fetal bovine serum (FBS) of different concentrations in the culture medium on osteogenic growth peptide (OGP) promoting bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation. MethodsBMSCs were separated from limb bones of 8 Sprague Dawley rats (5 weeks old) and purified by adherence method, and BMSCs at passage 3 were divided into 4 groups according to OGP concentration: OGP 1×10-10 mol/L group, OGP 1×10-9mol/L group, OGP 1×10-8 mol/L group, and control group without OGP; and 0, 2%, 5%, 8%, and 10%FBS concentration gradient was used in each group. The cell proliferation rate was detected by MTT method at 1, 3, 5, 7, 9, and 12 days after culture, and the activity of intracellular alkaline phosphatase (ALP) was determined by the method of p-nitrophenyl phosphate disodium at 9 days after culture. ResultsBMSCs showed adherent growth, rapid proliferation, long fiber vortex, and typical morphology. MTT analysis showed that cells could not sustain proliferation when FBS concentration was less than 5% in each group; when FBS concentration was above 8%, cells proliferated continually. Proliferation promoting effect of OGP 1×10-8 mol/L and 1×10-9 mol/L groups was significantly higher than that of the control group in all serum concentrations (P<0.05); when FBS concentration was lower than 10%, the proliferation promoting effect of OGP 1×10-8 mol/L group was significantly higher than that of the other 2 OGP groups (P<0.05), but when FBS concentration was 10%, OGP 1×10-8 mol/L group had no advantage of promoting proliferation. ALP test results showed that as the FBS concentration increased, ALP activity of all groups also significantly increased (P<0.05). Under the condition of 5%FBS and 8%FBS, the ALP activity of each OGP group was significantly greater than that of the control group, and it was the highest in OGP 1×10-8 mol/L group (P<0.05). Under the condition of 10%FBS, the ALP activity of each OGP group was still greater than that of the control group (P<0.05), but no significant difference was found between the OGP 1×10-8 mol/L group and OGP 1×10-9 mol/L group (P>0.05). ConclusionThe concentration of 8%FBS is the best concentration of serum for OGP promoting the proliferation and differentiation of BMSCs, and the most suitable concentration of promoting the proliferation and differentiation of BMSCs is OGP 1×10-8 mol/L.
ObjectiveTo evaluate the expression of miR-338-5p in colorectal cancer tissues and study its role in colon cancer cell proliferation, apoptosis, and cell cycle. MethodsThe expression of miR-338-5p was detected by real-time PCR in the colorectal cancer tissues and corresponding adjacent to cancer tissue samples. The miR-338-5p-mimics was transfected into the colon cancer cell lines HCT116 and SW620 to investigate its role in cell proliferation, apoptosis, and cell cycle. The cell proliferation and apoptosis were measured by CCK-8 and flow cytometry, respectively. The cell cycle was also analyzed by flow cytometry. Results①miR-338-5p expression was significantly downregulated in the colorectal cancer tissues as compared with corresponding adjacent to cancer tissue samples(P < 0.01). 2 Compared with the transfected negative control cells, the proliferation ability of colon cancer cell HCT116 or SW620 was significantly decreased(P < 0.01), cell apoptosis was significantly increased[HCT116 cell:(11.43±0.67)% versus(7.98±0.36)%, P < 0.01;SW620 cell:(10.5±0.2)% versus(7.93±0.5)%, P < 0.01), and cell G1 was arrested[HCT116 cell:(80.41±1.34)% versus (64.87±1.83)%, P < 0.01;SW620 cell:(68.76±0.41)% versus(54.89±0.78)%, P < 0.01) after transfecting miR-338-5p-mimics cells. ConclusionmiR-338-5p may act as an anti-oncogene in colorectal cancer through regulation of cell proliferation, apoptosis, and cell cycle.
ObjectiveTo understand the incidence of fecal incontinence after sphincter-preserving operation for middle and low rectal cancer, the factors influencing fecal incontinence, the relationship of fecal incontinence to anorectal manometry, and treatment. MethodThe literatures about fecal incontinence after sphincter-preserving operation for middle and low rectal cancer were reviewed. ResultsThe incidence of fecal incontinence after sphincter-preserving operation for middle and low rectal cancer was about 35.3%. Wexner fecal incontinence score was the most popular scale in assessing the severity of fecal incontinence, which had high validity and utility. When Wexner score≥9, the fecal incontinence-related quality of life was seriously damaged. Closer the anastomosis to the anal margin, the fecal incontinence was more likely to happen and much severer if it appeared. Surgeon could improve the anorectal function through some kinds of surgeries, like ultralow anterior resection with levator-sphincter reinforcement when the tumor site was rather low. The effect of chemoradiotherapy on fecal incontinence was uncertain now. Age itself was a risk factor for fecal incontinence, for elderly patients underwent sphincter-preserving operation needed to be careful. The relationship of fecal incontinence to anorectal function was not completely clear. The anal sphincter nerve function was a predicting factor whether neurogenic fecal incontinence was going to happen or not. Even though the retrograde colonic irrigation, sacral nerve stimulation, and biofeedback therapy had been proved to alleviate the symptoms and improve the quality of life after sphincter-preserving operation, much more prospective and controlled studies were needed to validate their efficacy and explore other new solutions. ConclusionsWe still need to come up with the objective criterion to assess fecal incontinence. Much more prospective studies are needed to analyze the influencing factors and to find effective prevention and treatment.
ObjectiveTo investigate the expression of catechol O-methyltransferase (COMT) mRNA and its protein in colorectal adenoma tissues and corresponding adjacent tissues, colorectal cancer tissues and corresponding adjacent tissues. MethodsExpressions of COMT mRNA and its protein were evaluated by real-time PCR and immunohistochemistry method in colorectal adenoma tissues and corresponding adjacent tissues, colorectal cancer tissues and corresponding adjacent tissues. Meanwhile, the relationship between the expression of COMT and clinic-pathological features of colorectal adenoma and colorectal cancer were analyzed. Results①The expression of COMT mRNA in colorectal adenoma tissue/colorectal cancer tissue group was higher than that of corresponding adjacent tissue group (0.109 0 vs. 0.000 5, t=3.02, P=0.01; 0.041 8 vs. 0.013 5, t=2.71, P=0.02).②The rate of high-expression of COMT in colorectal adenoma tissue/colorectal cancer tissue group was higher than that of corresponding adjacent tissue group [72.34% (34/47) vs. 25.53% (12/47), χ2=28.72, P < 0.01; 66.67% (28/42) vs. 28.57% (12/42), χ2=4.97, P < 0.05].③High-expression of COMT was not related to age, gender, location of tumor, and pathological type in colorectal adenoma patients (P > 0.05). High-expression of COMT was not related to age, gender, location of tumor, and differentiation degree (P > 0.05), but was related to TNM staging, T staging, and N staging in colorectal cancer patients (P < 0.05), the patients of TNMⅠ+Ⅱstaging, T1+T2 staging, and N0 staging had higher rate of high-expression of COMT. ConclusionCompared with corresponding adjacent tissues, COMT expresses highly in colorectal adenoma tissues and colorectal cancer tissues, so it may play a partial role in the emergence and development of colorectal cancer.