1.Screening of efficient siRNA target sites directed against gatekeeper genes for DNA repair.
Jinghua, REN ; Jusheng, LIN ; Xuyang, DONG ; Dong, XU ; Qiong, CHEN ; Yao, LIU ; Ying, CHANG ; Jinjian, YAO ; Siyuan, HAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):640-3
To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online, we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene. Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different, depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.
3.Protective effects of Platycodon grandiflorum total saponins on acute lung injury in rats and related mechanisms
Shuangyong DONG ; Yuansheng XU ; Yi WANG ; Xuyang ZHENG ; Wenbin SHENG
Chinese Journal of Emergency Medicine 2017;26(12):1413-1417
Objective To investigate the protective effects of Platycodon grandiflorum total saponins (PGTS) on acute lung injury (ALI) in rats and the related mechanisms.Methods Total of 60 SD rats were randomly (random number) divided into control group,model group,low-,middle-and high-dose PGTS group,and dexamethasone group,10 rats in each group.The latter 4 groups and dexamethasone group were injected with 50,100,200 mg/kg PGTS and 5 mg/kg dexamethasone,respectively.After 1 h,the latter 5 groups were intraperitoneally injected with mg/kg LPS to establish the ALI model.The clinical symptoms of the rats were observed.After 12 h,the arterial PaO2 and PaCO2,serum TNF-α and IL-10 level,lung wet/dry weight ratio (W/D),lung tissue SOD,GSH-Px and MDA level and NF-κB protein expression were determined.Results Rats in model group manifested noticeable symptoms of acute lung injury (ALI) and lung tissue lesions.In treatment group with appropriate PGTS dose,ALI symptoms and lung lesions were significantly alleviated,arterial PaO2 was markedly increased (P < 0.05),PaCO2 was decreased obviously (P < 0.05),serum TNF-α level was prominently decreased (P < 0.05),IL-10 level was strikingly decreased (P < 0.05),lung W/D ratio was significantly decreased (P <0.05),lung tissue SOD and GSH-Px level were distincdy increased (P <0.05),MDA was clearly decreased (P < 0.05),and NF-κB protein expression was plainly decreased (P < 0.05),compared with model group.Conclusions PGTS has undoubted protective effects on acute lung injury induced by LPS in rats.The mechanism may be associated with its role of anti-inflammation,anti-lipid peroxidation and down regulation of NF-κB protein level in lung tissue.
4.Effect of intra-articular injection of different concentrations of ozonated water on articular cartilage of rabbits with osteoarthritis
Xuyang XIAO ; Hengtao JI ; Zhen ZHANG ; Youlong ZHOU ; Zhigang ZHUANG ; Tieli DONG
Chinese Journal of Anesthesiology 2021;41(7):844-847
Objective:To evaluate the effect of intra-articular injection of different concentrations of ozonated water on articular cartilage of rabbits with osteoarthritis (OA).Methods:Twenty-four clean-grade New Zealand white rabbits of both sexes, weighing 2.0-3.0 kg, aged 6 months, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), OA group, low concentration ozonated water group (L group) and high concentration ozonated water group (H group). The OA model was established by intra-articular injection of papain.At 2 weeks after the model was successfully established, 10.0 and 20.0 μg/ml ozonated water 1.0 ml was injected into the knee joint of rabbits in L and H groups, respectively, and 0.9% sodium chloride solution 1.0 ml was injected once a week, 3 times in total in OA group.At 1 week after the last injection, the cartilage tissue of the knee joint was removed and stained with toluidine blue for evaluation of Mankin score (under light microscope). The activity of caspase-3 in chondrocyte was detected by enzyme-linked immunosorbent assay. Results:Compared with group C, the Mankin score and caspase-3 activity were significantly increased in the other 3 groups ( P<0.05). Compared with group OA, the Mankin score and caspase-3 activity were significantly decreased in group L and group H ( P<0.05). Compared with group L, the Mankin score was significantly increased, and the activity of caspase-3 was decreased in group H ( P<0.05). Conclusion:Injecting ozonated water 10.0 μg/ml and 20.0 μg/ml into the knee joint cavity both can inhibit the apoptosis in chondrocytes and reduce the damage to articular cartilage, however, high concentration of ozonated water can cause the denaturation of the articular cartilage matrix in rabbits with OA.
5.Screening of Efficient siRNA Target Sites Directed against Gatekeeper Genes for DNA Repair
Jinghua REN ; Jusheng LIN ; Xuyang DONG ; Dong XU ; Qiong CHEN ; Yao LIU ; Ying CHANG ; Jinjian YAO ; Siyuan HAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):640-643
To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online,we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene.Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different,depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.
6.Analysis on the nutritional composition of prepackaged foods commonly consumed by children aged 6-12 in Chengdu
XIAO Xuyang, WU Dong, TAN Bingbing, LYU Ruiqi, QIAO Tian, CHENG Guo
Chinese Journal of School Health 2023;44(4):517-520
Objective:
To analyze the current nutritional composition of commonly consumed prepackaged foods among children in Chengdu and to provide a scientific basis for health education among children and adolescents.
Methods:
Based on the 3 day and 24 hour dietary data of children aged 6-12 in Chengdu of the Southwest China Childhood Nutrition and Growth Cohort from 2021 to 2022, the nutritional information of prepackaged foods was collected by combining offline and online methods. All foods were classified step by step, and the nutrient content of each 100 g or 100 mL food was counted and graded.
Results:
A total of 1 902 children s prepackaged foods in 23 sub categories of 10 major categories were investigated. Nuts and seeds, snack foods, instant foods and other dairy products had higher total energy(2 476, 2 027, 1 728, 1 816 kJ/100 g), with the nutrient reference value percentage(NRV%) exceeding 20%. Fish, poultry, meat, eggs and their products had the highest protein content(22.8 g/ 100 g ) with an NRV% of 38%, nuts and seeds had the highest fat content(47.5 g/100 g) with an NRV% of 79%, confectionery and jelly had the highest carbohydrate content(82.1 g/100 g) with an NRV% of 27%, and seasoning flour products had the highest sodium content with an NRV% up to 118%. Seasoning flour products and instant foods were mostly high sodium, high fat and high carbohydrate food. Baked and puffed foods were almost high fat and high carbohydrate. Fish, poultry, meat, eggs, beans and their products were rich in protein but mostly high in sodium. Beverages and cold drinks were low in other nutrients except carbohydrate.
Conclusion
The nutrient content of various prepackaged foods commonly consumed by children in Chengdu are quite different. Most of the foods consumed by children are high sodium, high fat and/or high carbohydrate. Nutrition education should be strengthened to help children choose healthy foods.
7.Study on the risk factors of type 1 gastric neuroendocrine tumor in 123 cases of autoimmune gastritis
Mengge SU ; Xuyang DONG ; Xuexiu ZHANG ; Ya LI ; Haining ZHOU ; Xiangyu LI ; Jianning YAO
Chinese Journal of Digestion 2022;42(10):675-680
Objective:To investigate the risk factors of type 1 gastric neuroendocrine tumor (g-NET) in patients with autoimmune gastritis(AIG).Methods:From September 1, 2016 to February 28, 2022, 123 patients with AIG visited the First Affiliated Hospital of Zhengzhou University were retrospectively enrolled, including 37 cases with type 1 g-NET and 86 cases without type 1g-NET. The clinical data, serological indicators, and endoscopic manifestation of all the patients were analyzed, including the age at the time of AIG diagnosis (hereinafter referred to as the age at diagnosis), levels of gastrin 17 and pepsinogen Ⅰ (PGⅠ), presence or absence of gastric fundus and gastric body polyps, etc. The independent risk factors of type 1 g-NET in AIG patients were analyzed by univariate and multivariate logistic regression. The receiver operating characteristic curve (ROC) was plotted to analyze the optimal cut-off value, sensitivity and specificity of the independent risk factors in predicting type 1 g-NET in AIG patients. Independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:Compared with those of the AIG patients without type 1 g-NET, the age at diagnosis of AIG patients with type 1 g-NET was younger ((57.49±11.16) years old vs. (48.49±10.96) years old), the level of gastrin 17 was higher (200.21 ng/L, 121.85 ng/L to 244.40 ng/L vs. 244.40 ng/L, 182.50 ng/L to 248.02 ng/L), and the proportion of patients with gastric fundus and gastric body polyps was higher(18.6%, 16/86 vs. 56.8%, 21/37), and the differences were statistically significant( t=-4.13, Z=-3.06, χ2=17.90; P<0.001, =0.002 and <0.001). The results of univariate logistic analysis showed that the age at diagnosis ( OR=0.931, 95% confidence interval (95% CI)0.895 to 0.967), gastrin 17( OR=1.012, 95% CI 1.005 to 1.019), PGⅠ( OR=0.974, 95% CI 0.950 to 0.998)and gastric fundus and gastric body polyps( OR=5.742, 95% CI 2.461 to 13.399)were the influencing factors of type 1 g-NET in AIG patients ( P<0.001, =0.001, =0.033 and <0.001). The results of multivariate logistic regression analysis indicated that the age at diagnosis( OR=0.921, 95% CI 0.881 to 0.964), gastrin 17( OR=1.011, 95% CI 1.001 to 1.020), gastric fundus and gastric body polyps( OR=7.696, 95% CI 2.710 to 21.857)were the independent risk factors of type 1 g-NET in AIG patients ( P<0.001, =0.024 and <0.001). The results of ROC analysis demonstrated that the optimal cut-off values for the age at diagnosis and gastrin 17 in predicting type 1 g-NET were 56.50 years old and 206.40 ng/L, respectively; with sensitivity of 83.8% and 70.3%, respectively, and specificity of 54.7% for both ( P<0.001 and=0.003). Conclusion:The age at diagnosis< 56.50 years old, gastrin 17>206.40 ng/L and the presence of gastric fundus and gastric body polyps are independent risk factors of type 1 g-NET in AIG patients.
8.Progresses of functional MRI for exploring mechanism of neurovascular coupling changes in diabetes mellitus type 2
Dong YANG ; Shan XU ; Xuyang WANG ; Lina DU ; Lin LIN ; Jing SHEN ; Jianlin WU
Chinese Journal of Medical Imaging Technology 2024;40(1):125-129
The cognitive impairment of diabetes mellitus type 2(T2DM)is closely related to neurovascular coupling(NVC)changes,but the exact mechanism remains unclear.Functional MRI(fMRI)technology were able to jointly analyze NVC changes of T2DM,providing new ideas for revealing the mechanism of cognitive dysfunction caused by T2DM.The progresses of fMRI for exploring NVC changes in T2DM were reviewed in this article.
9.The clinical characteristics of immunocompetent adults with chronic active Epstein-Barr virus associated enteritis
Xuyang DONG ; Ji LI ; Yue LI ; Dong WU ; Yan ZHANG ; Wei CAO ; Weixun ZHOU ; Hong YANG ; Jiaming QIAN
Chinese Journal of Internal Medicine 2018;57(7):487-493
Objective To investigate the clinical and pathological characteristics of chronic active Epstein-Barr virus associated enteritis(CAEAE). Methods The clinical data of 6 CAEAE patients in Peking Union Medical College Hospital were retrospectively analyzed from January 2010 to November 2017, including clinical manifestations, endoscopic, pathological features, medications and clinical outcome. Results The male∶female ratio was 4∶2 and the average age was 34 years old. All patients did not have personal and family history of immunodeficiency. The common symptoms consisted of fever (6/6), abdominal pain (6/6), hematochezia (6/6) and diarrhea (5/6). The most frequently affected sites were colorectum (5/6), followed by small intestine (3/6). Further serologic tests revealed a high load of serum Epstein-Barr virus (EBV) DNA. The main manifestations under endoscopy were multifocal or diffuse irregular ulcers. There was inflammation in the ulcer bases and surrounding areas, where EBV-encoded small RNA (EBER) was positive by in situ hybridization (6/6). The common complications were massive hemorrhage (3/6) and intestinal perforation (2/6). None of treatment agents including glucocorticoid, ganciclovir, foscarnet sodium and cytotoxic drugs was effective. All patients died within 0.5 to 13 months after diagnosis. Conclusions CAEAE in immunocompetent individuals is a rare disorder with poor prognosis. It is difficult to differentiate CAEAE from inflammatory bowel disease due to similar clinical and endoscopic manifestations.
10.Clinical study of single kidney transplantation from young pediatric donors after death: 86 cases report
Xuyang LIU ; Xuyong SUN ; Jianhui DONG ; Ke QIN ; Haibin LI ; Song CAO ; Ying HUANG ; Meisi LI ; Liugen LAN ; Zhuangjiang LI ; Xiaocong KUANG ; Ning WEN ; Jixiang LIAO ; Zhao GAO ; Qingdong SU ; Dongge YANG
Chinese Journal of Organ Transplantation 2018;39(2):76-80
Objective By analyzing the perioperative management in our hospital to explore the clinical effect and safety of single kidney transplantation from deceased juveniles' donors.Methods We retrospectively analyze 86 cases of kidney transplantations from deceased juveniles' donors in our hospital from 2007 December to 2015 August.Results The success rate of the operations was 100%.The postoperative complications occurred as fellows:7 cases of acute rejection (8.14%);10 cases of drug intoxication (11.62%);21 cases of DGF (24.44%),4 cases of leakage of urine (4.65%),7 cases of lung infection (8.14%).Two cases (2.32%) died after the operation because of serious lung infection,and by corresponding treatment 47 cases recovered after 2-4 weeks.The creatinine level in 37 cases without any complications was 131.88 ± 44.20 μmol/L during discharge.Conclusion With strict selection,the organ from a deceased juvenile donor is safe and practicable.