BACKGROUND:Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰmaybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately.
OBJECTIVE:To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells.
METHODS:Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in celllines.
RESULTS AND CONCLUSION:The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successful y constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells.

Pylorus obstruction caused by foreign bodies in the gastric anrum and granulomatous inflammation is rarely seen.The clinical symptoms of this disease are unspecific.Combination of X-ray radiography,computed tomography and gastroscopy could make definite diagnosis.Differential diagnosis between pylorus obstruction and peptic ulcer,gastric cancer and duodenal obstruction should be done before operation.On April 13,2012,a patient with pylorus obstruction caused by foreign bodies in the gastric anrum and granulomatous inflammation was treated at the Second Hospital of Jiaxing,the imaging characteristics of the disease were summarized to provide referrence for the diagnosis and treatment of this disease.

Objective To summarize the computed tomography (CT) features of small intestinal obstruction caused by primary abdominal cocoon and investigate the essentials of diagnosis and differential diagnosis.Methods The retrospective descriptive study was adopted.The clinical data of 1 patient with small intestinal obstruction caused by primary abdominal cocoon who was admitted to the Second Affiliated Hospital of Jiaxing University on October 6,2014 were collected.The patient underwent abdominal CT on admission and at 10 hours after admission.The patient received emergency exploratory laparotomy after preoperative preparation,and then postoperative regular symptomatic treatment and pathological examination.Results of abdominal CT were observed,including imaging features of abdominal masses,extent of small intestinal obstruction,situation of intestinal tube within the masses,vessel distribution in the mesentery and fibrous capsules around the mesentery.Situation of operation,routine blood test,biochemical indicator,blood coagulation indexes,postoperative recovery,complications,results of pathological examination and situation of patient during follow-up were recorded.The follow-up by telephone interview and outpatient examination was applied to the patient till October 31,2015,including detecting the dietary,with or without symptoms of abdominal pain and distension,haematemesis and hematochezia,routine blood retest,liver function,renal function and CT.Results Results of abdominal CT on admission:(1) coronal plain scan of abdominal CT showed that there were signs of incomplete intestinal obstruction,and local small intestinal dilatation and gas accumulation in left abdominal region without specific sign.(2) Plain scan of abdominal CT showed that there were the coated sign with thickened fibrous capsules around the intestinal tube, banana shape ' with agglomerate and expanding-distortion intestinal tube and aggregative,stretching and twisting mesentery with abnormal vessel distribution.(3) Sagittal reconstruction images of abdominal CT showed that a huge mass consisted of fibrous capsules as cocoon and agglomerate and expanding-distortion intestinal tube was petal-like and fixed on posterior abdominal wall.(4) Coronal reconstruction images of abdominal CT showed that agglomerate and expanding-distortion intestinal tube was annularly surrounded by uneven thickness fibrous capsules with abnormal vessel distribution in the mesentery.Results of abdominal CT at 10 hours after admission:(1) coronal plain scan of abdominal CT showed that small intestinal obstruction was obviously exacerbated and expanding intestinal tubes were increased and aggravated.(2) Plain scan of abdominal CT showed that a typical sign of small intestinal obstruction was exacerbated and there were multiple air-fluid levels in the agglomerate and expanding-distortion intestinal tube with fluid and gas accumulation.Patient underwent successful enterodialysis + resection of fibrous capsules.During operation,a huge mass in the intestine from suspensory ligament of duodenum to ileocecum was fixed on posterior abdominal wall and surrounded by dense,smooth and white fibrous capsules,partial colon was also surrounded by fibrous capsules and greater omentum was missing.Patient received the postoperative supporting treatments of fasting,anti-infection and inhibition of acid.Number of white blood cells,absolute value and percentage of neutrophils,levels of high-sensitivity C-reactive protein and procalcitonin were 17.10 × 109/L,15.70 × 109/L,91.5％,127.49 mg/L and 1.370 μg/L by blood routine retest at postoperative day 1,respectively.Patient had normal liver,renal and coagulation functions.Fluid diet intake at postoperative week 1 was gradually replaced by normal diet intake.Patient had normal liver and renal functions by blood routine retest at postoperative day 10 and a good recovery without intestinal fistula,abdominal and pulmonary infections and other complications.Postoperative pathological examination showed that gross specimen was mainly composed of cocoon-shaped,grayish white and tough fibrous capsules.Fibrous capsules were consisted of proliferative fibrofatty tissues by microscope observation,with small vascular hyperplasia and large numbers of the inflammatory cell infiltration.Patient was diagnosed with small intestinal obstruction caused by primary abdominal cocoon after operation,and followed up for 1 year with normal diet intake and without abdominal pain and distension,nausea and vomiting,melena and discomfort.There was normal blood routine retest,liver and renal functions and distribution of the intestine in abdomen by CT examination.No dilatation of the intestinal tube was found and strip-like high density shadow in ileocecum was detected and considered as remnant fibrous capsules.Conclusion Imaging features of small intestinal obstruction caused by primary abdominal cocoon include agglomerate and expanding-distortion intestinal tube fixed on abdomen,partial intestinal tubes dilatation,intestinal fluid accumulation and air-fluid level,aggregative,stretching and twisting mesentery with abnormal vessel distribution,thickened fibrous capsules around intestinal loops and among intestinal tubes.

Objective:To investigate the influence of circ_BACH2 on the malignant biological behavior of papillary thyroid cancer and its molecular mechanism.Methods:Cancer tissues and paracancer tissues of 51 patients with papillary thyroid carcinoma from the Fourth Central Hospital of Tianjin between 2017 and 2019 were collected. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expressions of circ_BACH2, miR-370-3p and G protein coupled receptor kinase interacting factor 1 (GIT1) mRNA in tissues and cells; flow cytometry to detect cell apoptosis and cell cycle; plate clone formation experiment to detect the number of cell clones; cell counting kit 8 (CCK-8) to detect cell proliferation; Transwell array to detect cell migration and invasion; western blot to detect protein expressions; dual luciferase report experiment to detect the targeting relationship between circ_BACH2, miR-370-3p and GIT1; the nude mouse tumor formation experiment to detect the effect of circ_BACH2 on tumors in mice.Results:Compared with adjacent tissues, the expressions of circ_BACH2 and GIT1 in papillary thyroid cancer tissues was increased, while the expression of miR-370-3p was decreased. Compared with Nthy-ori3-1 cells, the expressions of circ_BACH2 in papillary thyroid cancer cells TPC-1 and SW579 were increased, the mRNA and protein levels of GIT1 were increased, miR-370-3p expression was decreased. The expression level of GIT1 mRNA was negatively correlated with that of miR-370-3p ( r=-0.634), and the expression level of circ_BACH2 was positively correlated with that of GIT1 ( r=0.635). The expression level of circ_BACH2 was negatively correlated with that of miR-370-3p ( r=-0.394, P＜0.05). Circ_BACH2 and miR-370-3p has a binding site at the 3' UTR of GIT1. After knocking down circ_BACH2, the proportion of G 0/G 1 cells in papillary thyroid cancer cells TPC-1 and SW579 was increased, the proportion of S-phase cells was decreased and the proportion of G 2/M-phase cells did not change significantly. The cell absorbance value was lower than that in si-NC group. The number of cell clone formation was decreased (43±5 vs 100±6, 54±8 vs 100±9); the cell apoptosis rate was increased [(19.60±2.40)% vs (4.30±0.20)%, (18.10±2.10)% vs (5.10±0.23)%]; cell migration number was decreased (61±7 vs 134±15, 58±6 vs 112±11), the invasion number was also decreased (45±6 vs 113±11, 47±4 vs 92±9); the expressions of Snail and Twist1 were decreased, and the expression of E-cadherin was increased ( P＜0.000). Inhibition of miR-370-3p expression reversed the effect of circ_BACH2 knockdown on proliferation, migration, invasion and apoptosis of thyroid papillary cancer cells. Overexpression of GIT1 reversed the effects of overexpression of miR-370-3p on proliferation, migration, invasion and apoptosis of thyroid papillary cancer cells. Mice injected with TPC-1 cells stably transfected with sh-circ_BACH2 showed a reduction in tumor volume [(535±91) mm 3 vs (857±114) mm 3] after 35 days of culture; tumor weight was decreased [(0.62±0.13) mg vs (1.06±0.15) mg, P＜0.05]; the expressions of circ_BACH2 and GIT1 were decreased, and the expression of miR-370-3p was increased in nude mouse tumor tissue. Conclusion:Silencing circ_BACH2 may inhibit the proliferation, migration and invasion of papillary thyroid cancer cells in vitro, promote cell apoptosis, and inhibit tumor growth in vivo through targeted regulation of miR-370-3p/GIT1.

Objective:To investigate the influence of circ_BACH2 on the malignant biological behavior of papillary thyroid cancer and its molecular mechanism.Methods:Cancer tissues and paracancer tissues of 51 patients with papillary thyroid carcinoma from the Fourth Central Hospital of Tianjin between 2017 and 2019 were collected. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expressions of circ_BACH2, miR-370-3p and G protein coupled receptor kinase interacting factor 1 (GIT1) mRNA in tissues and cells; flow cytometry to detect cell apoptosis and cell cycle; plate clone formation experiment to detect the number of cell clones; cell counting kit 8 (CCK-8) to detect cell proliferation; Transwell array to detect cell migration and invasion; western blot to detect protein expressions; dual luciferase report experiment to detect the targeting relationship between circ_BACH2, miR-370-3p and GIT1; the nude mouse tumor formation experiment to detect the effect of circ_BACH2 on tumors in mice.Results:Compared with adjacent tissues, the expressions of circ_BACH2 and GIT1 in papillary thyroid cancer tissues was increased, while the expression of miR-370-3p was decreased. Compared with Nthy-ori3-1 cells, the expressions of circ_BACH2 in papillary thyroid cancer cells TPC-1 and SW579 were increased, the mRNA and protein levels of GIT1 were increased, miR-370-3p expression was decreased. The expression level of GIT1 mRNA was negatively correlated with that of miR-370-3p ( r=-0.634), and the expression level of circ_BACH2 was positively correlated with that of GIT1 ( r=0.635). The expression level of circ_BACH2 was negatively correlated with that of miR-370-3p ( r=-0.394, P＜0.05). Circ_BACH2 and miR-370-3p has a binding site at the 3' UTR of GIT1. After knocking down circ_BACH2, the proportion of G 0/G 1 cells in papillary thyroid cancer cells TPC-1 and SW579 was increased, the proportion of S-phase cells was decreased and the proportion of G 2/M-phase cells did not change significantly. The cell absorbance value was lower than that in si-NC group. The number of cell clone formation was decreased (43±5 vs 100±6, 54±8 vs 100±9); the cell apoptosis rate was increased [(19.60±2.40)% vs (4.30±0.20)%, (18.10±2.10)% vs (5.10±0.23)%]; cell migration number was decreased (61±7 vs 134±15, 58±6 vs 112±11), the invasion number was also decreased (45±6 vs 113±11, 47±4 vs 92±9); the expressions of Snail and Twist1 were decreased, and the expression of E-cadherin was increased ( P＜0.000). Inhibition of miR-370-3p expression reversed the effect of circ_BACH2 knockdown on proliferation, migration, invasion and apoptosis of thyroid papillary cancer cells. Overexpression of GIT1 reversed the effects of overexpression of miR-370-3p on proliferation, migration, invasion and apoptosis of thyroid papillary cancer cells. Mice injected with TPC-1 cells stably transfected with sh-circ_BACH2 showed a reduction in tumor volume [(535±91) mm 3 vs (857±114) mm 3] after 35 days of culture; tumor weight was decreased [(0.62±0.13) mg vs (1.06±0.15) mg, P＜0.05]; the expressions of circ_BACH2 and GIT1 were decreased, and the expression of miR-370-3p was increased in nude mouse tumor tissue. Conclusion:Silencing circ_BACH2 may inhibit the proliferation, migration and invasion of papillary thyroid cancer cells in vitro, promote cell apoptosis, and inhibit tumor growth in vivo through targeted regulation of miR-370-3p/GIT1.