Objective To investigate the expression of urokinase-type plasminogen activator (uPA) mRNA in gastric cancer tissues and cancer-adjacent tissues and the relationship between its expression and biologic behavior of tumor. Methods Fourty-eight cases with gastric cancer were detected for the expression of uPA mRNA by fluorogenic probe quantitative reverse transcription polymerase chain reaction (RT-PCR). Results The positive expression rate of uPA mRNA was 83.3%, 25.0%, 93.8% and 62.5% in gastric cancer tissues,cancer-adjacent tissues, gastric cancer tissues with lymph node metastasis and with non-lymph node metastasis respectively. Expression of uPA mRNA was positively related with the invasion depth of gastric cancer.Conclusion Expression of uPA mRNA is significantly increased in gastric cancer and it can be used as an indicator to judge the metastasis and prognosis of tumor.

AIM:To investigate the depressant effect and mechanism of atorvastatin on the chronic rejection of aortic allograft in rats.METHODS:The models of abdominal aorta transplantation were made with micro-surgery in rats.The recipients were divided into three groups:allograft control group,atorvastatin-treated group and isograft control group.Vascular intimal thickness in all of the groups was observed by histological examination.The expression of proliferation cell nuclear antigenl(PCNA)and ?-smooth muscle actin(?-SMA)were determined by immunohistochemistry.The content of nitric oxide was measured by nitrate reductase chromatometry.RESULTS:The vascular intimal thickness in atorvastatin-treated group(11.60%?2.40%)was lower than that in allograft control group(34.60%?6.40%,P

Objective: The impact of parenteral fish oil lipid emulsion on liver function and nutritional status of malignant tumors of the liver and gallbladder patients. Methods: From December 2007 to A-pril 2008, 32 post-operative hepatobiliary cancer patients were randomly divided into control and study groups. Two groups were treated with isocaloric, isonitrogenic parenteral nutrition and the study group was added fish oil lipid emulsion. Comparison of plasma protein, glucose, jaundice index, transaminase, ALP and the rate of infection complications was made betweent the two groups. Results: The blood glucose, transaminase and ALP levels were not significantly different between the two groups. But the plasma proteins and bilirubin levels were improved significantly (P < 0.05) with reduced infection complication in the study group. Conclusion : Fish oil lipid emulsion is conducive to the recovery of post-operative liver and gallbladder cancer patients in live function and nutritional status.

AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.

The eukaryotic expression of human arresten gene and its effect on the proliferation of in vitro cultured vascular smooth cells (VSMCs) in vitro were investigated. COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome. Forty-eight h after transfection, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of arresten mRNA in the cells, while Western blot assay was applied to detect the expression of arresten protein in concentrated supernatant. Primary VSMCs from thoracic aorta of male Sprague-Dawley rats were cultured using the tissue explant method, and identified by immunohistochemical staining with a smooth muscle-specific anti-alpha-actin monoclonal antibody before serial subcultivation. VSMCs were then co-cultured with the concentrated supernatant and their proliferation was detected using Cell Counting Kit-8 (CCK-8) in vitro. The results showed that RT-PCR revealed that the genome of arresten-transfected cells contained a 449 bp specific fragment of arresten gene, suggesting the successful transfection. Successful protein expression in supernatants was confirmed by Western blot. CCK-8 assay showed that the proliferation of VSMCs were inhibited significantly by arresten protein as compared with control cells (F=40.154, P<0.01). It was concluded that arresten protein expressed in eukaryotic cells can inhibit proliferation of VSMCs effectively in vitro, which would provide possibility to the animal experiments.

In order to investigate the origin of neointimal smooth muscle cells in transplant arteriosclerosis in rat aortic allograft, sex-mismatched bone marrow transplantation was performed from male Wistar rats to female Wistar rats. Four weeks after transplantation, the aortic transplant model was established by means of micro-surgery in rats. The recipients were divided into 4 groups: female Wistar-female Wistar aortic isografts, female SD female Wistar aortic allografts, male SD-male Wistar aortic allografts, female SD-chimera Wistar aortic allografts. Eight weeks after transplantation, aortic grafts were removed at autopsy and processed for histological evaluation and immunohistochemistry. The results indicated that excessive accumulation of alpha-SMA-positive smooth muscle cells resulted in significant neointima formation and vascular lumen stricture in rat aortic allografts. Neointima assay revealed that the neointimal area and NIA/MA ratio of transplanted artery were significantly increased in all of aortic allograft groups as compared with those in aortic isograft group (P<0.01). Neointimal smooth muscle cells were harvested from cryostat sections of aortic allograft by microdissection method. The Sry gene-specific PCR was performed, and the result showed that a distinct DNA band of 225 bp emerged in the male-male aortic allograft group and chimera aortic allograft group respectively, but not in the female-female aortic allograft group. It was suggested that recipient bone-marrow cells, as the origin of neointimal smooth muscle cells, contributed to the pathological neointimal hyperplasia of aortic allograft and transplant arteriosclerosis.

Objective To study the depressant effect of atorvastatin on arteriosclerosis of aortic allograft in rats.Methods The models of abdominal aorta transplantation were established in rats with the use of(micro-surgery).The recipients were divided into three groups:allograft control group,allograft experimental group and isograft control group.After 60 days of transplant,vascular intimal thickness(VIT) in all of the groups was observed by histological examination.The expression of PCNA and ?-SMA was determined by(immunohistochemistry).Results The degree of VIT in rats of the allograft experimental group was lower than that in the allograft control group;the VIT area ratio in the allograft control group,allograft experimental group and isograft control group was(12.40?2.65)%,(5.20?6.35)%,and(1.2?1.10)%,respectively,A statistical difference between these groups was observed(P

AIM: To express human Arresten gene in eukaryotic cell,and to investigate its effect on the proliferation and migration in vitro of rat primary cultured thoracic aortic vascular smooth cells (VSMCs).METHODS: COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome.48 hours after transfection,polymerase chain reaction(RT-PCR) was used to detect the expression of Arresten mRNA in the cells,while Western blotting assay was applied to detect expressed Arresten protein in concentrated supernatants.VSMCs were then co-cultured with the concentrated supernatants;and its proliferation was detected using cell counting kit-8(CCK-8) in vitro.Migration of VSMCs was assayed by a microchemotaxis chamber and a polycarbonate filter (Transwell's chamber) with pores of 8 ?m in diameter.RESULTS: RT-PCR revealed that the genome of Arresten-transferred cells contained a 449bp specific fragment of Arresten gene.Successful protein expression in supernatants was confirmed by Western blotting.CCK-8 assay showed that the proliferation of VSMCs was inhibited significantly by Arresten protein as compared with control group(P

To express recombinant arresten in Escherichia coli (E. Coli) and investigate its biological activity, prokaryotic expression vector of human arresten gene was constructed by gene engineering. Human arresten gene was amplified from recombinant plasmid pGEMArr by polymerase chain reaction (PCR), and inserted into prokaryotic expression vector pRSET containing T7 promoter. Restriction analysis and DNA sequencing verified that the arresten gene was correctly cloned into the expression vector. The recombinant plasmid pRSETAt was subsequently transformed into E. Coli BL21 (DE3), and the target gene was expressed under induction of IPTG. SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 29 kD (1 kD=0. 992 1 ku) amounted to 29 % of the total bacterial proteins. After purification and renaturation, the recombinant protein could significantly suppress the proliferation of human umbilical vein endothelial cells (HUVECs). These results suggested that the expression of a biologically active form of human arresten in the pRSET expression system laid a foundation for further study on the mechanistic insight into arresten action on angiogenesis and the development of powerful anti-cancer drugs.

Objective:To explore the optimal match degree between thoracolumbar kyphosis (TLK) and lower lumbar lordosis (LLL) in adult spinal deformity (ASD) after correction surgery.Methods:Data of 119 ASD patients (male: 28, female: 91), belonging to the Affiliated Hospital of Jining Medical University (19 cases), the Affiliated Hospital of Shandong University of Traditional Chinese Medicine (11 cases), and the First Medical Center of Chinese PLA General Hospital (89 cases) were reviewed and documented from March 2019 to March 2020. All patients (age, 64.48±8.88 years; range, 45-79 years) underwent the surgical procedure of thoracolumbar fusion with instrumentations were followed up over 24 months (51.68±15.60 months; range, 24-87 months) after surgery. Postoperative proximal interface failure, Oswestry disability index (ODI) score and Scoliosis Research Society-22 (SRS-22) score were recorded for all patients. The immediate match of TLK to LLL postoperatively was calculated as follows: TLM=TLK/LLL. The data of those individuals with excellent improvements in the ODI (>50%) at the final follow-up were recorded and analyzed. Then the mean value and the 95% CI of TLM in those individuals were calculated. All participants were subdivided into three groups according to the 95% CI value of TLM. After the receiver operating characteristic curve (ROC) analyzing, the area under the ROC curve (AUC) was the best cutoff value of TLM. The association of proximal junctional failure (PJF) developing with the abnormal TLM postoperatively was analyzed with logistic regression, and the odds ratio (OR) was calculated. Results:62 patients had significant improvements in ODI (>50%) at the final follow-up, and the mean TLM in those individuals was 0.41 [95% CI (0.2, 0.5)]. All patients were divided into three groups: TLM<0.2 (35 cases), 0.2≤TLM≤0.5 (48 cases) and TLM>0.5 (36 cases). The preoperative TLK (13.87°±16.61°) and T 1 pelvic angle (19.69°±10.55°) in the those patients with TLM<0.2 were the smallest, and those were the largest in those with TLM>0.5 (30.59°±16.68°, 28.30°±14.46°). The individuals with TLM<0.2 still had the smallest TLK (2.89°±1.78°), however, those with TLM>0.5 had the largest TLK (17.13°±12.13°) and the smallest LLL (-26.16°±11.02°) accordingly. Additionally, the ODI and SRS-22 for those with 0.2≤TLM≤0.5 at the final follow-up were the best ( P<0.05). ROC curve analysis results showed that the best cutoff value of TLM was 0.4 (sensitivity=78.9%, specificity=76.2%; AUC=0.802, 95% CI (0.708, 0.896) , P<0.001). During the follow-up after orthopedic surgery, there were 19 patients with postoperative proximal junction failure, including 16 patients in the mismatched group (6 patients in the TLM<0.2 group, 10 patients in the TLM>0.5 group) and 3 patients in the matched group (0.2≤TLM≤0.5 group), with the incidence of 23% (16/71) and 6% (3/48), respectively. The difference was statistically significant (χ 2=5.66, P=0.017). Thoracolumbar mismatch was significantly associated with proximal borderline failure after orthosis [ OR=4.35, 95% CI (1.196, 15.924)]. Conclusion:The abnormal correction in thoracolumbar kyphosis and lower lumbar lordosis may result in mismatch between thoracolumbar segments, which would undermine the quality of life, and increase the incidence of proximal junctional failure developing in those ASD patients underwent long-fusion surgeries. The match between TLK and LLL should be 0.2 to 0.5.