1.cDNA cloning and sequence analysis of ribosomal protein S7 gene from giant panda
Yiling HOU ; Chunlian WU ; Wanru HOU ; Yanzhe HAO ; Tian ZHANG
Journal of Third Military Medical University 2003;0(18):-
Objective To explore the structure of ribosomal protein S7 ( RPS7) gene of giant panda ( Ailuropoda melanoleuca) and investigate its homologies with other already reported sequences,including Homo sapiens,Mus musculus,Rattus norvegicus and Bos taurus. Methods The cDNA of RPS7 was cloned from the giant panda by RT-PCR. The sequence data were analyzed by GenScan software. Blast 2. 1 was used to study the homology of the obtained RPS7 sequence with the gene sequences of other species; Open reading frame ( ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the RPS7 sequence cloned was deduced using Predict Protein software. Results The full length of the sequence fragment was 589 bp containing an ORF of 585 bp. The deduced protein sequence showed that the protein was composed of 194 ami- no acids and its estimated molecular weight was 22. 126 85 ?103 with a pI of 10. 09. There were 7 different pat- terns of functional sites: one N-glycosylation site; two cAMP and cGMP-dependent kinase phosphorylation sites; four casein kinase C phosphorylation sites; one casein kinase Ⅱ phosphorylation site; two N-myristoylation sites; two amidation sites and one ribosomal protein S7e signature site in the RPS7 protein. Further analysis indicated that the sequence of RPS7 and the protein encoded were highly homologous to some mammals reported.Conclusion The complete coding sequence of RPS7 gene has been cloned through RT-PCR successfully, which is the first report on the RPS7 gene from the giant panda.
2.Ischemic precondition inhibits pancreatic acinar cells apoptosis in rats with ischemia/reperfusion injury following pancreas transplantation: Role of reactive oxygen and mitochondrial DNA repair enzyme
Yiling HOU ; Hai BO ; Ziquan LIU ; Shihai XIA
Chinese Journal of Tissue Engineering Research 2010;14(18):3279-3285
BACKGROUND: Ischemic preconditioning (IPC) can induce endogenous protection mechanism, which effectively prevent ischemia/reperfusion injury following organ transplantation. Cold and warm ischemia may induce ischemia/reperfusion injury of pancreas transplantation, and apoptosis of pancreatic acinar cells is one of the important reasons of pancreas graft functional defect after transplantation. Mitochondrial DNA has repair system, and its balance with mitochondrial DNA injury influences disease occurrence and outcome.OBJECTIVE: To observe the effect of IPC on apoptosis of transplanted pancreatic acinar cells, and the possible role of reactive oxygen (ROS) and mitochondrial DNA repair enzyme.METHODS: A total of 50 health, male, Sprague-Dawley rats were randomly divided into three groups: sham operated (n = 10), donors (n = 20) and recipients (n = 20). The recipients were randomly divided into ischemia/reperfusion group (IR, n = 10) and IPC group (n = 10). The sham operated group was subjected to abdominal open and close operation. IR group and IPC group received establishment of diabetic model by streptozotocin injection. IR rats received whole pancreatic-duodenal transplantation alone. IPC rats received whole pancreatic-duodenal transplantation exposed ischemic preconditioning with 5 minutes ischemia and 5 minutes reperfusion twice. All grafts were keep with warm ischemia time 15 minutes and cold ischemia (in 4 ℃ UW preservation solution) time 180 minutes. Twelve hours after reperfusion, serum amylase, blood glucose, Caspase-3, -9 activity were detected. Pancreatic acinar cell apoptosis was measured by flow cytometry. Mitochondrial cross-membrane potential (Δψ) was measured by monitoring the fluorescence spectrum of rhodamine 123. Mitochondrial H2O2 generation was determined using dichlorofluorescein as a probe. 8-oxodG in mitochondrial DNA (mtDNA) was measured with HPLC system. Release of cytochrome C, phosphorylation of Akt and mitochondrial OGG1 protein expression were determined by Western-blotting. RESULTS AND CONCLUSION: The ischemia preconditioning can relieve the pancreatic acinar cell apoptosis in pancreas graft and relieve IR injury by decreasing mitochondrial oxidative stress, mtDNA injury, and increasing phosphorylation of Akt and mitochondrial OGG1 expression.
3.Hydrogen-rich water inhibits mitochondrial oxidative stress and inflammation in the skeletal muscle after eccentric exercise
Lei WANG ; Ziquan LIU ; Yiling HOU ; Yaojun GE
Chinese Journal of Tissue Engineering Research 2015;(29):4682-4687
BACKGROUND:It is unclear whether hydrogen-rich water can be used to protect skeletal muscle injury induced by eccentric exercise, as wel as the relative mechanism. OBJECTIVE:To observe the effect of hydrogen-rich water on the mitochondrial oxidative stress and inflammation in rat skeletal muscle after eccentric exercise, and to investigate the relative signaling pathway of hydrogen-rich water. METHODS:Forty Sprague Dawley rats were randomly divided into four groups: control group, eccentric exercise group, eccentric exercise+saline group, and eccentric exercise+hydrogen-rich water group. Rats in three eccentric exercise groups were exercised on a motor-driven rodent treadmil at a speed of 16-18 m/min and a slope of-16° for 90 minutes per day. Rats in the eccentric exercise+hydrogen-rich water group were subjected to intraperitoneal injection of hydrogen-rich water (10 mL/kg) immediately after exercise; and rats in the eccentric exercise+saline group were administrated with normal saline after exercise. Al the interventions lasted for 5 days. RESULTS AND CONCLUSION:Hydrogen-rich water intervention after eccentric exercise could markedly enhance the mitochondrial Sirtuin-3 expression, improve the mitochondrial membrane potential and activity of manganese superoxide dismutase, down-regulate the mitochondrial reactive oxygen species generation and mitochondrial DNA oxidative damage, thus inhibiting inflammatory cytokines expression, such as NLRP3 and interleukin-1β. The results indicated that hydrogen-rich saline could directly scavenge reactive oxygen species. In addition, hydrogen-rich water could improve mitochondrial energy metabolism and antioxidant capacity through up-regulation of Sirtuin-3, which in turn inhibits eccentric exercise-induced mitochondrial oxidative stress and secondary inflammation in the skeletal muscle.
4.Tertiary Rehabilitation in Military Sanatorium
Qinghua CHEN ; Shouqin SHAN ; Fanggao HOU ; Xinnian DAI ; Meiyan PAN ; Yiling LIU
Chinese Journal of Rehabilitation Theory and Practice 2010;16(3):300-300
Based on the resources of military sanatorium, we developed a mode of rehabilitation that combined the hospital-, sanatorium- and community-based rehabilitation as a whole.
5.Study on the role of oxidative stress in the kidneys of diabetic rats
Xiaobo LI ; Zhongqing MU ; Li CHEN ; Yiling FU ; Yan ZHANG ; Yuxin XIAN ; Xinguo HOU
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To investigate the changes of oxidative stress in the kidneys and their roles in nephropathy in diabetic rats. METHODS: Diabetic rats were induced by streptozotocin (STZ). 36 rats were divided into three groups randomly: (1) NC group, normal control rats; (2) DM group, diabetic rats received protamine zinc insulin (PZI) 2U-4U/2 d; (3) DT group, diabetic rats received PZI 9-12 U/kg body weigh/day. 12 weeks later, rats were killed, blood glucose, blood cholesterol, serum creatinine, blood urea nitrogen, HbA1c, urinary creatinine, and urinary protein for 24 h were measured. The activities of antioxidant enzymes in renal cortex, including total superoxide dismutase (TSOD), Cu-Zn superoxide dismutase (Cu-Zn SOD), Mn superoxide dismutase (Mn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and maleic dialdehyde (MDA) were measured by chromatometry. RT-PCR was performed to detect the expression of different antioxidant enzymes mRNA. RESULTS: For all the targets we measured, there was no significant difference between NC and DT groups. Compared with the other two groups, the levels of blood glucose, cholesterol, trigalloyl glycerol, HbA1c in DM group increased significantly. The activities of TSOD, Cu-Zn SOD and CAT decreased significantly. The activity of GSH-Px increased significantly. There was no significant difference among the activities of Mn SOD in all three groups. The level of MDA in DM group was much higher than that in NC or DT group. The relative expression levels of GSH-Px and Cu-Zn SOD mRNA in DM group were higher than those in other two groups, while the relative expression level of CAT decreased. Mn SOD mRNA was expressed without significant difference in all groups. Compared with NC or DT group, urinary protein in DM group increased significantly, while creatinine clearance rate decreased. CONCLUSIONS: Hyperglycemia affected the expression of antioxidant enzymes. Oxidative stress was caused by hyperglycemia in diabetic rats and may be an important factor in the etiology of diabetic nephropathy.
6.Effects of two different tranexamic acid administration methods on perioperative blood loss in total hip arthroplasty: study protocol for a prospective, open-label,randomized, controlled clinical trial
Zhenyang HOU ; Yiling SUN ; Tao PANG ; Dong LV ; Biao ZHU ; Zhen LI ; Xingyu CHAI ; Zhengwen XU ; Changzheng SU
Chinese Journal of Tissue Engineering Research 2017;21(15):2314-2319
BACKGROUND: Reducing perioperative blood loss in total hip arthroplasty is a hot topic for joint surgeons. Both intravenous infusion and intra-articular injection of tranexamic acid significantly reduce perioperative blood loss, blood transfusion volume, and need for blood transfusion in patients undergoing total hip arthroplasty. However, differences between the intravenous and intra-articular methods are not clear.OBJECTIVE: To evaluate the effects of these two tranexamic acid administration methods on perioperative blood loss in patients undergoing total hip arthroplasty.METHODS: We are conducting a prospective, single-center, open-label, randomized, controlled clinical trial at the Tengzhou Central People's Hospital, China. Ninety patients undergoing unilateral total hip arthroplasty have been randomized into three groups. In the intravenous infusion group (n=30), 15 mg/kg tranexamic acid diluted in 100 mL physiological saline was infused intravenously at the beginning of surgery and 20 mL of physiological saline was injected intra-articularly after deep fascia suturing. In the intra-articular injection group (n=30), 100 mL of physiological saline was infused intravenously at the beginning of surgery and a mixture of 1.5 g tranexamic acid and 20 mL physiological saline was injected intra-articularly after deep fascia suturing. In the control group (n=30), 100 mL of physiological saline was infused intravenously at the beginning of surgery and 20 mL of physiological saline was injected intra-articularly after deep fascia suturing. The primary outcome is hidden blood loss at 1 and 3 days postoperatively. The secondary outcomes are visible blood loss, need for blood transfusion, and mean blood transfusion volume intraoperatively and on days 1 and 3 postoperatively. Other outcomes are the incidence of adverse reactions and complications within 3 months of surgery. The study protocol has been approved by the Ethics Committee of Tengzhou Central People's Hospital of China, approval number 2015-026. All protocols will be performed in accordance with the Ethical Principles for Medical Research Involving Human Subjects in the Declaration of Helsinki. Written informed consent was provided by each patient and their family members after they indicated that they fully understood the treatment plan.DISCUSSION: This trial was designed in April 2015. Cases were collected in July 2015. Data analysis will be finished in December 2017. This study is designed to investigate the effects of intravenous infusion versus intra-articular injection of tranexamic acid on perioperative blood loss in patients undergoing total hip arthroplasty to determine the more effective mode of administration.
7. Study on preventive and therapeutic effect of Tongbu-fangchan prescription on Ach-CaCl
Lu LIU ; Ya-Fen WANG ; Zhen-Hua JIA ; Lu LIU ; Ya-Fen WANG ; Zhen LI ; Zhen-Hua JIA ; Yu-Jie YIN ; Yun-Long HOU ; Deng-Feng XU ; Zhen-Hua JIA ; Yu-Jie YIN ; Yun-Long HOU ; Deng-Feng XU ; Zhen-Hua JIA ; Yu-Jie YIN ; Yun-Long HOU ; Deng-Feng XU ; Zhen-Hua JIA
Chinese Pharmacological Bulletin 2023;39(7):1385-1392
Aim To explore he preventive and therapeutic effects of Tongbu-fangchan prescription on aceylcholine-calcium chloride (Ach-CaCl
8.Identification Study on Cryptotympana pustulata Ecdysis Based on 3D Depth of Field Synthesis Technology
Lixiao GUO ; Zijing XUE ; Fangjie HOU ; Baohui SUN ; Yuguang ZHENG ; Qian ZHENG
China Pharmacy 2020;31(4):447-451
OBJECTIVE:To establish the rapid field identification method of Cryptotympana pustulata ecdysis. METHODS : 3D depth of field synthesis technology was used to identify 50 batches of C. pustulata ecdysis and its adulterants from the length of beak ,size and protrusion degree of upper labial base ,the protrusion degree of the lower labial base ecdysis and the color of its upper transverse groove ,the number and shape of main and lateral spines on the foot ,significance of abdominal valves ,the number of webs ,the number and shape of side plates ,the number of tergum rings ,terminaliae,etc. RESULTS :Among 50 batches of samples ,S1-S5,S26-S30,S36-S50 were C. pustulata ecdysis;S21-S25 was adulterants of C. pustulata ecdysis after weight gain ;S31-S35 was adulterants of C. pustulata ecdysis after extraction ;S6-S20 were ecdysis from Tibicen flammatus ,C. flammatta,Lyristes pekinensis ,all of which were adulterants. The main distinguishing feature of C. pustulata ecdysis and its adulterants was that abdomen and ventral surface of C. pustulata ecdysis were triangular ,and the abdomen and ventral surface of other species was nearly parallel ;the valve of C. flammatta ecdysis was obvious ,but those of other varieties were not obvious ;the lateral appearance of terminaliae of C. flammatta ecdysis was sharper than those of other species ;there was an acute angle between the front foot accessory thorns and the end thorns of the T. flammatus ecdysis,and an obtuse angle between the front foot accessory thorns and the end thorns of the L. pekinensis ecdysis. CONCLUSIONS :The method is simple ,reliable and suitable for rapid field identification of C. pustulata ecdysis and its adulterants.
9.Repositioning of clinically approved drug Bazi Bushen capsule for treatment of Alzheimer's disease using network pharma-cology approach and in vitro experimental validation
Tongxing WANG ; Meng CHEN ; Bin HOU ; Junqing LIANG ; Cong WEI ; Zhenhua JIA
Chinese Journal of Pharmacology and Toxicology 2023;37(z1):22-23
OBJECTIVE To explore the new indications and key mechanism of Bazi Bushen capsule(BZBS)by network pharmacology and in vitro experiment.METHODS The potential tar-get profiles of the components of BZBS were pre-dicted.Subsequently,new indications for BZBS were predicted by disease ontology(DO)enrich-ment analysis and initially validated by GO and KEGG pathway enrichment analysis.Further-more,the therapeutic target of BZBS acting on AD signaling pathway were identified by intersec-tion analysis.Two Alzheimer's disease(AD)cell models,BV-2 and SH-SY5Y,were used to pre-liminarily verify the anti-AD efficacy and mecha-nism of BZBS in vitro.RESULTS In total,1499 non-repeated ingredients were obtained from 16 herbs in BZBS formula,and 1320 BZBS targets with high confidence were predicted.Disease enrichment results strongly suggested that BZBS formula has the potential to be used in the treat-ment of AD.In vitro experiments showed that BZ-BS could significantly reduce the release of TNF-α and IL-6 and the expression of COX-2 and PSEN1 in A β 25-35-induced BV-2 cells.BZBS reduced the apoptosis rate of A β 25-35 induced SH-SY5Y cells,significantly increased mitochon-drial membrane potential,reduced the expres-sion of Caspase3 active fragment and PSEN1,and increased the expression of IDE.CONCLU-SIONS BZBS formula has a potential use in the treatment of AD,which is achieved through regu-lation of ERK1/2,NF-κB signaling pathways,and GSK-3β/β-catenin signaling pathway.Further-more,the network pharmacology technology is a feasible drug repurposing strategy to reposition new clinical use of approved TCM and explore the mechanism of action.The study lays a foun-dation for the subsequent in-depth study of BZBS in the treatment of AD and provides a basis for its application in the clinical treatment of AD.
10.Bazi Bushen Capsules on Anti-aging: A Review
Hong-rong LI ; Cong WEI ; Chuan-yuan JI ; Xin-jing MAO ; Dan-dong WANG ; Kun MA ; Jing AI ; Yun-long HOU ; Zhen-hua JIA
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(7):239-246
The decline of body function and senile diseases caused by aging seriously affect human health and life span, which is an important topic in the field of life science. Bazi Bushen capsules is a representative Chinese patent medicine for tonifying essence, invigorating Qi and anti-aging, guided by Qiluo doctrine, and essence, Qi and spirit theory. Previous pharmacological and clinical studies have confirmed that this preparation has the comprehensive advantages of anti-aging, and prevention and treatment of aging-related diseases. Among them, pharmacological studies showed that Bazi Bushen capsules had the effect of improving the appearance status of mice, improving the level of sex hormones, inhibiting the formation of atherosclerosis, improving cardiac function, improving learning and memory cognitive ability, improving neurological function, improving osteoporosis and muscle function, improving sperm count and quality. The mechanism was related to the up-regulation of the recombinant sirtuin (SIRT6) level, down-regulation of the levels of aging-related proteins p53 and p16, up-regulation of telomerase reverse transcriptase level, and alleviation of inflammation and oxidative response. Clinical studies have proved that it can improve the symptoms of patients with kidney essence deficiency, improve exercise ability, and improve the sexual function of impotence patients. Anti-aging research of Bazi Bushen capsules based on Qiluo doctrine fully embodies the new mode of academic innovation and transformation of traditional Chinese medicine (TCM) with the combination of "theory-new drug-experiment-clinic", which has made a demonstration for the anti-aging research of TCM.