Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
Animals ; Cloning, Molecular ; Gene Expression Regulation ; Genetic Vectors ; Mice ; NIH 3T3 Cells ; Promoter Regions, Genetic ; Superoxide Dismutase ; genetics ; Swine ; Transfection

Objective:To evaluate the cytotoxicity of colorized zirconia ceramics.Methods:Zirconia ceramic 3Y-TZP blocks were colorized by NH4 VO3 ,FeCl3 ,Co(NO3 )3 ,Ni(NO3 )3 and MoCl3 respectively.The cytotoxicity of colorized zirconia to L-929 cells was examined by immersion method and MTT assay.Results:During 3 days culture,L929 cells in all groups showed similar prolifera-tion and normal morphology.The toxicity gradation of all groups was 0 -I.Conclusion:Zirconia ceramics colorized with the 5 kinds of colorants has no cytotoxicity.

Objective To investigate the impact of preload fasting and meal replacement in patients with metabolic syndrome.Methods A total of 92 subjects with metabolic syndrome were enrolled in the study.They were assigned into the preload fasting group (PFG),the meal replacement group (MRG),and the control group (CG) for 12-weeks intervention.Special dietary with 100 kcal was provided 30 min before each meal in the PFG,and while in the MRG the same dietary was taken just before each meal and the amount of meal was reduced appropriately.The subjects in CG took meals as usual.Body mass index,waist circumference,and insulin resistance were assessed.Satiety situation was investigated by the scale.Results After 12 weeks,improvement were found in fasting insulin(-3.29 mU/L) and waist circumference (-4.04 cm) in the PFG and significant difference was shown compared to the CG (P＜0.05).Satiety index in the PFG was the most significant among the three group.Conclusion Preload fasting is helpful in improving insulin resistance,reducing waist circumference,and enhancing satiety.

Objective The aim of this study was to investigate the relationship between phosphotyrosine interac-tion domain containing 1 ( PID1 ) gene and variation in intramuscular fat ( IMF ) content and the possibility to generate transgenic animals by testicular injection .Methods Expression vector pIRES2-acGFP-PID1 carrying pig PID1 gene was incubated with transfection reagents and injected into the testes of male New Zealand rabbits .We examined the F1 genera-tion by fluorescence detection , PCR, Western blotting and measuring the IMF content .The F1 generation gave birth to the F2 generation.Then we examined the F2 generation through detecting the positive rate and the IMF content .Results The exogenous PID1 gene and fluorescent protein gene were expressed at different levels both in the F 1 generation and the F2 generation, and the positive rates were 35.88%and 34.33%, respectively.The IMF content was significantly elevated (P<0.05) in the transgenic positive individuals compared with the negative ones and the control group , and the PID1 protein expression was similarly higher .Conclusions The results of this study demonstrate that PID 1 gene affects intramuscular fat content significantly .Moreover, the results of our analysis provide further evidence that transgenic animals can be gener -ated by testicular injection , and the exogenous gene can be inherited steadily .

Objective To evaluate the influence of dosage,operation method,adverse reaction of endoscopic photodynamic therapy (EPDT) on its therapeutic efficacy in rabbit models of in-situ rectal cancer,so as to provide preclinical basis of photodynamic therapy for rectal cancer.Methods 20 rabbits with in-situ VX2 rectal cancer were randomly divided into control group,PDT low dose group,intermediate dose group,and high dose group.At 24 h before PDT,photosensitizer (hermimether) was intravenously injected into rabbits.630 nm semiconductor laser was used as light source.The growth of the tumor was observed by conventional endoscopy and endoscopic ultrasonography,and the survival time,general conditions and adverse reactions were recorded.The histopathological changes were observed by hematoxylin-eosin staining.Results At 7 d after PDT,the total response rates of low dose,intermediate dose and high dose group respectively were 40％ (slight),80％ (60％ remarkable and 20％ slight),100％ (20％ remarkable and 80％ slight).The average survival times of the three groups were 14 d,10 d and 5 d,respectively.The main adverse reactions were inflammation,intestinal obstruction,intestinal peristalsis loss and death.Conclusions The dosage of PDT is an important factor to influence the curative effect.The appropriate dose of PDT will have a better effect on the treatment of rectal cancer.A thorough study of these problems is helpful to the clinical application of PDT in the treatment of rectal cancer.

Objective:Colorectal cancer screening was performed on a general population with age ranging between 40 and 74 years old to evaluate the screening effects of questionnaire survey, fecal occult blood (FOB) test, and colonoscopy, as well as to provide some implications of colorectal cancer screening strategies. Methods: Two-step screening model of questionnaire survey combined with FOB test was applied for the screening. Colonoscopy was conducted in a high-risk population identified through preliminary screening as final diagnosis. Results:Based on the 2,117,304 cases screened, the screening compliance was 39.72%, and 126,118 cases (5.96%) were identified as high risk. Colonoscopies were performed on 25,837 cases, of which 8,095, 1,236, 134, 112, and 336 were identified as adenoma, advanced adenoma, severe dysplasia lesions, early cancer, and advanced cancer, respectively. The early stage di-agnostic rate was 81.52%. Conclusion:The colorectal cancer screening method performed in Tianjin can significantly concentrate on the high-risk population with colorectal cancer, increase the positivity rate of total colonoscopy, and economize medical resources.

Objective To evallhte factors affecting blood loss during mini-percutaneous nephrolithotomy using ureteroscope and pneumatic intracorporeal lithotripsy(MPCNL). Methods 1156MPCNL procedures in 885 patients from July 2002 to October 2006 were reviewed. Various patientrelated and intraoperative factors were assessed for association with total blood loss using multivariate regression analysis. ResuIts The average hemoglobin drop was(14.2+8.3)g/L.The overall blood transfusion rate was 1.5%. Approximately 0.6%of patients required angiography embolization to control intractable bleeding. Multivariate regression analysis showed that the occurrence ot operative complications(6=0.496,P＜0.001),size of the tract(b=0.405,P＜0.001),mature nephrostomy tract(6=0.377,P＜0.001),multiple tracts(6=0.326,P=0.005),size of stone(b=0.210,P=0.015),operative time(6=0.139,P=0.027),renal parenehymal thickness(b=0.128,P=0.035),prior stone intervention(b=-0.121,P=0.038),diabetes(b=0.110,P=0.051),and bacteriuria(b=-0.095,P=0.058)were significant predictors of blood loss.Factors such as age,sex,side,obesity,hypertension,renal function,operating surgeon,anaesthesia,calix of puncture,number of attempts to successful puncture,urine from puncture needle may not affect the blood loss. Concluslons Maneuvers that may reduce blood loss include reducing the operative time,decreasing the occurrence of intraoperative complications, reducing the tract size and tract number, and staging the procedure.Staging the procedure of MPCNL is a judicious decision in case of a large stone burden,intraoperative complications,multiple-tract,lager tract or diabetes.

Objective To evaluate the feasibility and curative effect of extracorporeal liver perfusion (ECLP) in treatment of acute liver failure (ALF) in pigs. Methods The experiments were carried out in healthy pigs (weight 20-30 kg) under general anesthesia. All of the pigs were randomly divided into 3 groups. ALF model was established by liver blood supply obstructing and portal-systemic shunting. ALF group (n=5): ALF pigs were killed 8 h after establishing. ALF+ECLP group (n=5): ALF pigs were perfused with ECLP for 4 h and killed 8 h after establishing. Normal liver+ECLP group (n=4): normal pigs were dealt with just as ALF+ECLP group. The data of PT, AST, TNF, blood ammonia were collected in all groups. Pathologic changes in liver and brain were detected. Results The levels of PT, AST, TNF, blood ammonia, RBC and HCT in the ALF+ECLP group were lower than those in the ALF group (P

Objective Acute aortic dissection occurs when a tear in the inside of the aorta causes blood to flow between the layers of the wall of the aorta,forcing the layers apart.In most cases this is associated with a sudden onset of severe chest or back pain,often described as "tearing" in character.The main management includes medication,endovascular repair and surgery.Medical management plays an very important role in the management of acute aortic dissection.Aortic dissection generally presents as a hypertensive emergency.Individuals can benefit from blood pressure control and anti-impulse therapy in perioperative period.

Objective:To investigate the RHD genotypes of RhD-negative pregnant women and explore the optimum strategy for fetal RHD screening among this population in the region. Methods:This prospective study recruited 33 cases of RhD-negative singleton pregnancies at ≥12 weeks of gestation in Nanjing Drum Tower Hospital from March to November 2021. On the basis of RHD genotyping, quantitative real-time polymerase chain reaction (PCR) was used to amplify the exons 5 and 10 of RHD gene in the circulating cell-free DNA of RhD-negative pregnant women harboring whole RHD gene deletion and RHD-CE(2-9)- D. High-throughput sequencing was performed to detect chr1:25648453 locus from circulating cell-free DNA in plasma of RhD-negative pregnant women harboring RHD 1227A mutation to screen the fetal RhD blood group. Neonatal umbilical cord blood samples were collected for verifying fetal RHD genotyping. Descriptive statistical analysis was used. Results:Whole RHD gene deletion homozygous genotype ( n=20, 60.6%), RHD-CE(2-9) -D/whole RHD gene deletion heterozygous genotype ( n=5, 21.2%), RHD 1227A/whole RHD gene deletion heterozygous genotype ( n=7, 15.2%) and RHD 711delC/whole RHD gene deletion heterozygous genotype ( n=1) were identified in the 33 RhD-negative pregnant women. In the 25 cases with whole RHD gene deletion homozygous genotype or RHD-CE(2-9)- D/whole RHD gene deletion heterozygous genotype, 22 fetuses were RhD-positive and three were RhD-negative based on prenatal screening, which were confirmed by the neonatal serological test results after birth. In the seven cases carrying RHD 1227A/whole RHD gene deletion heterozygous genotype, all fetuses were RhD-positive, which were consistent with the results of serological detection after delivery. The case harboring RHD 711delC/whole RHD gene deletion heterozygous genotype did not receive fetal RHD screening. Conclusions:This study suggests that whole RHD gene deletion homozygous genotype is the most common allele in RhD-negative population in this area, followed by RHD 1227A/whole RHD gene deletion heterozygous genotype and RHD- CE(2-9)- D/whole RHD gene deletion heterozygous genotype. For women with whole RHD gene deletion homozygous genotype, RHD- CE(2-9)- D, or RHD 1227A mutation, fetal RHD screening with quantitative real-time PCR and high-throughput sequencing are important for the management of RhD-negative pregnant women.