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Objective:To investigate the expression and clinical significance of growth differentiation factor 15 (GDF-15) in papillary thyroid carcinoma (PTC).Methods:The tumor tissues and metastatic lymph node tissues of 3 PTC patients who underwent radical surgery of thyroid cancer in the First Hospital of Hunan University of Chinese Medicine from January to February 2019 were collected, and the differential expressed genes were screened by high-throughput sequencing; 20 cases of primary tumor tissues and metastatic lymph node tissues were collected to verify the sequencing results. Another 20 cases of primary PTC tissues and adjacent tissues (>2 cm away from the tumor edge) were collected to verify the expression of target genes in tumor tissues and adjacent tissues. Sixty-four pathological specimens of PTC patients who underwent radical surgery of thyroid cancer in the First Hospital of Hunan University of Chinese Medicine from January to December 2014 were collected, of which 31 patients had lymph node metastasis. The real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of GDF-15 and verify the sequencing results; immunohistochemistry was used to detect the expression of GDF-15 protein in the primary PTC tissues, adjacent tissues and metastatic lymph node tissues. According to the expression of GDF-15 protein in the primary tumor tissues of PTC patients, the patients were divided into high-expression group (35 cases) and low-expression group (29 cases), and the relationship between GDF-15 expression level and clinicopathological characteristics of patients was analyzed; Kaplan-Meier method was used to analyze the 5-year tumor-free survival rate of the two groups.Results:The mRNA high-throughput sequencing results of 3 cases of PTC primary and metastatic tissues showed that the top 10 differential expressed genes were CDH2, CDF15, DKK1, GLIPR1, PCDH7, ID3, FBN1, MYPN, UBASH3B and CCDC80. The expression of GDF-15 mRNA in 20 cases of PTC primary tumor tissues and adjacent tissues were 4.1±0.5 and 2.8±0.3, and the difference was statistically significant ( t = 2.220, P = 0.032). The expression of GDF-15 mRNA in another 20 cases of PTC primary tumor tissues and metastatic lymph node tissues were 3.1±0.4 and 5.8±0.7, and the difference was statistically significant ( t = 3.556, P = 0.001). The results of immunohistochemistry showed that GDF-15 had the immunohistochemical scores of (4.0±0.3) points, (6.1±0.3) points and (9.0±0.4) points in PTC adjacent tissues, primary tumor tissues and metastasis tissues. The expressions of GDF-15 protein between PTC primary tumor tissues and adjacent tissues, metastatic tissues and adjacent tissues, and metastatic tissues and primary tumor tissues were significantly different (all P < 0.01). The differences in composition ratios of tumor long-axis diameter, tumor T stage and N stage between GDF-15 high-expression group and low-expression group were statistically significant (all P<0.05). The 5-year tumor-free survival rates in GDF-15 high-expression group and low-expression group were 60% and 83%, and the difference was statistically significant ( P = 0.033). Conclusions:The expressions of GDF-15 in PTC adjacent tissues, tumor tissues and metastatic lymph node tissues gradually increase, and its expression level is related to tumor progression, recurrence and metastasis. It can be used as a potential clinical prognostic warning molecule and therapeutic target.
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Objective:To investigate the relationship between the expression of inhibin subunit Beta A (INHBA) and the clinicopathological data and its effect on proliferation, invasion and metastasis of gastric cancer cells and its possible mechanisms.Methods:Using Gene Expression Profiling Interactive Analysis (GEPIA) public database to verify the expression of INHBA mRNA in gastric cancer and adjacent tissues and its relationship with pathological stage and prognosis; the relationship between INHBA and clinical prognosis was analyzed using the Kaplan-Meier Plotter online database. Immunohistochemistry was used to confirm the correlation between protein expression level of INHBA in gastric cancer and adjacent tissues and clinical pathological staging. In vitro, the cell proliferation was detected by tetrazolium salt (MTT) method; the cell migration ability was detected by scratch test, and cell invasion and metastasis ability was detected by transwell chamber assay. The expression of INHBA and epithelial-mesenchymal transition (EMT) related proteins was detected by Western blot. Results:The GEPIA database and Kaplan-Meier Plotter online database analysis showed that INHBA mRNA was highly expressed in various cancer tissues and significantly higher in gastric cancer tissues than normal tissues ( P<0.05). The high expression of INHBA mRNA was associated with clinical stage and poor prognosis of gastric cancer ( P<0.05). The results of immunohistochemistry showed that the staining score of INHBA in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.05), and its expression level was correlated with clinical tumor node metastasis (TNM) stage ( P<0.05). The results of MTT, scratch test and transwell chamber showed that INHBA overexpression could promote the proliferation, migration, invasion and metastasis of gastric cancer cells, while interference with INHBA expression could inhibit the proliferation, migration, invasion and metastasis of gastric cancer cells; Western blot results showed that the expression of CDH1 was down regulated and the expression of CDH2 was up-regulated after INHBA overexpression. The expression of CDH1 was up-regulated and the expression of CDH2 was down-regulated after INHBA overexpression was inhibited. Conclusions:INHBA is highly expressed in gastric cancer tissues compared with adjacent tissues. The expression level of INHBA is related to tumor progression and poor prognosis. INHBA can promote the proliferation, migration and invasion of gastric cancer MGC-803 cell line and its mechanism may be related to INHBA promoting cell EMT.
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Objective:To investigate the effects of microRNA (miR)-513a-3p regulating hexokinase 2 (HK2) on proliferation and glycometabolism in colorectal cancer cell.Methods:From May 2019 to February 2020, the miR-513a-3p simulant, miR-513a-3p inhibitor and miR control were transfected into colorectal cancer SW480 cell respectively. Real-time quantitative polymerase chain reaction was used to detect the expression levels of miR-513a-3p in colorectal cancer SW480 cell, normal colorectal cell and all transfected colorectal cancer SW480 cell. The effect of miR-513a-3p on cell proliferation was detected by CCK-8 assay. Brd/PI incorporation assay was used to detect the effect of miR-513a-3p on glycometabolism. Western blot was used to detect the expression of HK2.Results:The expression level of miR-513a-3p in colorectal cancer SW480 cell was significantly lower than that in normal colorectal cell (0.43 ± 0.06 vs. 1.00 ± 0.02), and there was statistical difference ( t = 7.024, P = 0.003). The expression level of miR-513a-3p in colorectal cancer SW480 cell transfected with miR-513a-3p simulant was significantly higher than that in colorectal cancer SW480 cell transfected with miR control and transfected with miR-513a-3p inhibitor (1.18 ± 0.24 vs. 0.45 ± 0.04 and 0.22 ± 0.03), the expression level of miR-513a-3p in colorectal cancer SW480 cell transfected with miR control was significantly higher than that in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor, and there were statistical differences ( P<0.05). The proliferation ability in colorectal cancer SW480 cell transfected with miR-513a-3p simulant at 24, 48, 72 and 96 h was significantly lower than that in colorectal cancer SW480 cell transfected with miR-513a-3p control group, the proliferation ability in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor at 24, 48, 72 and 96 h was significantly higher than that in colorectal cancer SW480 cell transfected with miR-513a-3p control, and there were statistical differences ( P<0.05). The glucose intake, lactate and thioredoxin-interacting protein (TXNIP) expression levels in colorectal cancer SW480 cell transfected with stimulant were significantly lower than those in colorectal cancer SW480 cell transfected with miR control (1.02 ± 0.04 vs. 1.90 ± 0.06, 0.88 ± 0.03 vs. 1.45 ± 0.04 and 0.16 ± 0.02 vs. 0.86 ± 0.06), the indexes in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor were significantly higher than those in colorectal cancer SW480 cell transfected with miR control (2.35 ± 0.09 vs. 1.90 ± 0.06, 1.67 ± 0.08 vs. 1.45 ± 0.04 and 2.01 ± 0.15 vs. 0.86 ± 0.06), and there were statistical differences ( P<0.05). The expression level of HK2 in colorectal cancer SW480 cell transfected with miR-513a-3p stimulant was significantly lower than that in colorectal cancer SW480 cell transfected with miR control (0.20 ± 0.01 vs. 1.02 ± 0.04), and there was statistical difference ( t = 8.367, P<0.05); the expression level of HK2 in colorectal cancer SW480 cell transfected with miR-513a-3p inhibitor was significantly higher than that in colorectal cancer SW480 cell transfected with miR control (1.91 ± 0.07 vs. 1.02 ± 0.04), and there was statistical difference ( t = 4.279, P<0.05). Conclusions:MiR-513a-3p can significantly inhibit the proliferation and glycometabolism of colorectal cancer cell, and its regulatory mechanism is related to the inhibition of the HK2 protein expression in the cell by miR-513a-3p.
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Objective:To evaluate the pedicled bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap for the treatment of soft tissue defects at the contrallateral leg.Methods:Between January of 2012 and January of 2016, 8 patients with soft tissue defects at the leg were treated at Department of Orthopedic Surgery, Hand and Foot Surgery Hospital of Lanzhou. They were 5 men and 3 women, aged from 19 to 50 years (mean, 35 years). All of them were treated by bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap pedicled with posterior tibial artery. The size of the defects ranged from 10 cm×9 cm to 13 cm×8 cm. The immediate coverage of the muscle flaps and vessel pedicle was repaired by a meshed split-thickness skin graft. The donor site was closed directly. The therapeutic efficacy was assessed at the final follow-up according to the criteria by Iowa for tibial fractures.Results:All the skin flaps and muscle flaps survived without any vascular crisis. One case developed necrosis of small skin graft at the distal muscle flap which spontaneously healed after dressing change for 2 weeks. Their follow-up ranged from 2.5 to 4.5 years (mean, 3.8 years). A good contour was confirmed at the recipient area. By the Iowa criteria at the final follow-up, 3 cases were excellent, 4 good and one fair.Conclusion:Pedicled bridge transplantation with medial leg skin flap and medial hemi-soleus muscle flap is a good treatment for soft tissue defects at the contrallateral leg which has only one major blood vessel, reducing damage to the donor site.
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Objective:To investigate the expression and clinical significance of oxidized low density lipoprotein receptor 1 (OLR1) in gastric cancer.Methods:In gastric cancer tissues and adjacent tissues, the relationship between the expression of OLR1 and the stage and clinical prognosis of gastric cancer was analyzed from the Gene Expression Profiling Interactive Analysis (GEPIA) database and Kaplan-Meier Plotter online database. We collected 6 case of fresh gastric cancer tissues and adjacent tissues of patients undergoing radical gastrectomy in our hospital from October 2018 to December 2018, The extracted RNA were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) to verify the expression in gastric cancer and adjacent tissues. From January 2012 to January 2014, 36 patients with pathological biopsy specimens of radical gastrectomy in our hospital were examined. Immunohistochemistry was used to confirm the correlation between protein expression level of OLR1 in gastric cancer and paracancerous tissues and clinical pathological stage and prognosis.Results:GEPIA database and Kaplan-Meier Plotter online database analysis showed that OLR1 mRNA was highly expressed in various cancer tissues and significantly higher in gastric cancer tissues than normal tissues ( P<0.05). High expression of OLR1 mRNA is associated with clinical stage and poor prognosis of gastric cancer; qRT-PCR results showed that OLR1 mRNA was highly expressed in gastric cancer tissues; the results of immunohistochemistry showed that the staining score of OLR1 in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.001), and its expression level was correlated with depth of tumor invasion and clinical tumor node metastasis (TNM) stage (both P<0.05). Kaplan-Meier survival analysis showed that the expression level of OLR1 was associated with poor prognosis in patients. The median survival time was 18 months in OLR1 high expression group and 30 months in OLR1 low expression group, with statistically significant difference ( P=0.016). Conclusions:OLR1 is highly expressed in gastric cancer tissues, and its expression level is associated with tumor progression and poor prognosis.
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Objective Triple negative breast cancer(TNBC), a special breast cancer subtype, is lack of effective target therapy.The article aimed to investigate the role of lysophosphatidic acid (LPA) and Hippo Yes-associated protein (Hippo-YAP) signaling pathway in TNBC invasion and metastasis and the mechanisms.Methods The specific small interfering RNA (siRNA) of YAP was synthetized in vitro, and was transfected into MDA-MB-231 cells using liposome transfection.The experiment was divided into YAP-siRNA group, positive control group and blank control group.Each group is provided with 2 parallel holes.Evaluation was made on the effects of each group on Hippo-YAP, the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group, YAP content, the capacity of invasion and metastasis after transfection ([0.035±0.005], [2.200±1.000], [3.500±0.800]) significantly decreased compared with positive control group([0.343±0.012], [27.600±5.100], [22.300±5.000]) and blank control group([0.384±0.017], [26.500±4.800], [22.350±6.000]) (P<0.05).YAP expression levels at 60 min, 120 min, and 240 min in experiment group significantly decreased compared with positive control group and blank control group (P<0.05).YAP relative expression levels of 10, 20, 50 μmol/Lwere significantly lower than those of positive control group and blank control group (P<0.05).After respective interference of C3 transferase and Y27623, significant difference was found in the pYAP mRNA contents of experiment group([0.255±0.052], [0.326±0.017]), blank control group([0.048±0.032], [0.534±0.017]) and positive control group([0.052±0.021], [0.528±0.024])(P<0.05).The expression levels of YAP mNA and AREG mNA significantly increased in experiment group([0.176±0.032], [0.263±0.008]) compared with blank control group([0.043±0.013], [0.263±0.008]) and positive control group([0.049±0.025], [0.057±0.043])(P<0.05).Conclusion LPA induces breast cancer invasion and metastasis, which is YAP-dependent, time-dependent and concentration-dependent.LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.
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Objective To evaluate the differential expression of lysophosphatidic acid receptor (LPAR) in breast cancer (BC), and its relationship with clinicopathological features of BC patients. Methods The qRT-PCR and immunohistochemical staining were used to detect the LPAR expression in 37 normal tissues, 55 benign disease tissues and 82 BC tissues, besides, the correlation of LPAR expression with clinicopathological data was also analyzed. Results The expression levels of LPAR2 and LPAR3 mRNA and protein in BC tissues were higher than those in normal benign tissues (all P0.05). LPAR1 expression was not associated with clinicopathological features in BC tissues (P>0.05). LPAR2 expression in postmenopausal patients was higher than that in premenopausal patients (χ2=4.821, P<0.05). LPAR3 expression was significantly associated with nodal metastasis, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) in BC tissues (all P<0.05). Conclusion LPAR in BC tissues has differential expression, which is associated with nodal metastasis, ER, PR and HER2.
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Objective To study the local anatomic structure of subclavian vein and have a good command of operating methods and skills on subclavian vein catheterization (SVC) so as to improve the success rate of puncture. Methods Retrospectively analyze the clinical data of 600 cases of hospitalized patients who have had SVC (male 410, female 190) in our hospital from March 2010 to March 2014. Results There were 582 cases (97%) of successful puncture,including 546 cases (91%) of one-time puncture success,38 cases (6. 3%) of repeated puncture or other types of puncture,and 16 cases (2. 7%) of failed puncture. Postoperative complications occurred in 22 patients (3. 7%),including 8 cases (1. 3%) of strayed into the artery,3 cases (0. 5%) of pneumothorax,5 cases (0. 8%) of catheter tip into the internal jugular vein,2 cases (0. 3%) of catheter related infection,2 cases (0. 3%) of catheter blockage,and 2 cases (0. 3%) of arrhythmia. Conclusion SVC is easy,safe and reliable,and it is of high success rate and less complications. Catheter retention time of SVC is longer. From the above,SVC is a good central venous puncture method which is worth of popularizing in clinical use.
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<p><b>OBJECTIVE</b>To optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow (BMSCs).</p><p><b>METHODS</b>BMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages.</p><p><b>RESULTS</b>The cells isolated using this method were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages. The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing hepatocyte- and cholangiocyte-specific markers.</p><p><b>CONCLUSION</b>The optimized method allows efficient isolation of homogenous populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.</p>
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Animals , Rats , Cell Culture Techniques , Cell Differentiation , Cell Proliferation , Cell Separation , Flow Cytometry , Mesenchymal Stem Cells , Cell BiologyABSTRACT
Objective To compare the efficiency of repairing perimembranous ventricular septal defect (VSD) with continuous stitching to that with interrupted stitching. Methods Of the 158 patients with perimembranous VSD ( 54% males) ,102 ( 65% ) patients underwent the repairing of continuous stitching (continuous group) ,and 56 patients underwent interrupted stitching (interrupted group). All the surgeries were performed under hypothermic cardiopulmonary bypass. Results The cardiopulmonary bypass time and aortic cross-clamp times were (31 ±14) mins and (18 ±12) mins in the continuous group, which was significantly less that those of (42 ± 16) mins and (25 ±11) mins in the interrupted group (t =4.49 and 3. 61 .respectively ,P <0. 05) . No case was died. The main complications involved temporary M atrioventricular block (AVB) of two cases (1 case in each group),which disappeared after 1 week. There were two remnants remaining leak (n =2) which was less than 3 mm and automatically cured after 3-6 months. Complete right bundle branch block(CRBBB) occurred in 3 cases in the continuous group and 6 cases in the interrupted group,of which 3 disappeared after 1 month and 2 disappeared after 12 months. The outcome was good in all cases after 1-3 years follow up. Conclusions Continuous stitching showed a shorter cardiopulmonary bypass and aortic cross-clamp times, and reduced the CRBBB.
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Objective To evaluate the clinical efficacy of transcatheter arterial chemoembolization (TOCE) and percutaneous absolute ethanol injection (APEI) in moderate or advanced liver cancer.Method During May,1993～Aug,1999, 137 patients with unresectable advanced liver cancer were retrospectively studied. 85 cases only received TOCE, 52 cases were treated by TOCE+APEI by B-US guidance.Results The effect in TOCE+APEI group was significant better than that in only TOCE group,the half year, 1-year,2-year and 3-year survival rates of the TOCE+APEI group were 86 54%,67 31%,43 55% and 21 77%,while those of the TOCE group were 65 89%,47 92%,22 12% and 1 48% respectively. There was significant difference between the two groups in survival rate(P
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Objective To explore the effect of hepatectomy or cholangiolithotomy on patients with hepatocholangic stones. Methods The clinical data of surgical treatment of 316 patients with hepatocholangic stones combined with bile duct stricture was retrospectively analyzed. The patients were divided into two groups:172 patients chiefly underwent hepatectomy,and the other 144 patients chiefly underwent cholangiolithotomy. Results The ratio of residual stones and reoperation in the hepatectomy group was significantly lower than those in the cholangiolithotomy group (P