1.Report of 5 gene-edited pig-rhesus monkey heterotopic heart xenotransplantation experiment
Gen ZHANG ; Huan WANG ; Yulong GUAN ; Jie YAN ; Ji LI ; Xiaoliang LI ; Xianhua LI ; Rong ZHOU ; Xianzhi WANG ; Zhipeng REN ; Dongsheng HE ; Xin LI ; Dengke PAN ; Dianyuan LI
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(6):379-384
Objective:To investigate the changing trends in cardiac function following xenogeneic heterotopic heart transplantation of multi-gene edited pig hearts and assess the impact of recipient immune responses on donor heart, laying experimental groundwork for the clinical application of gene editing technology.Methods:On December 16, 2023, xenogeneic heterotopic heart transplantation was performed between pigs and rhesus monkeys. Functional status of the graft under post-transplantation load conditions and recipient immune indicators were observed.Results:The recipient monkeys survived for 40 days with satisfactory functionality of both donor and recipient hearts, and no hyperacute or acute immune rejection reactions were observed.Conclusion:Multi-gene editing technology provides potential for xenotransplantation, yet further exploration is needed for its clinical application.
2.Chinese yam yield is affected by soil nutrient levels and interactions among N, P, and K fertilizers.
Yang CHEN ; Xianzhi ZHOU ; Lina MA ; Yongsheng LIN ; Xiangui HUANG
Chinese Herbal Medicines 2023;15(4):588-593
OBJECTIVE:
The current fertilization methods for Chinese yam are uneconomic and unfriend to environment. A rational one is very important to achieve desired balance of high yield of Chinese yam, economic and friend to environment. Here, we studied the effects of nitrogen (N), phosphorus (P), and potassium (K) fertilizers on the yield of 'Qinfeng' Chinese yam in shallow-groove directional cultivation.
METHODS:
The experiments were conducted in Dehua County, Fujian Province, China using a "3414" optimal design. Overall, three fertilizer factors (N, P, and K) were evaluated at the following four levels: 0, no fertilizer; 1, 0.5-fold the typical rate; 2, typical fertilization rate; and 3, 1.5-fold the typical rate. There were 14 different fertilization treatments.
RESULTS:
Treatment 6 (N2P2K2) produced the longest (75.6 cm) and thickest tubers (4.9 cm) with the highest tuber fresh weight (1311.9 g) and yield (41 015.9 kg/hm2), whereas, treatment 1 produced the shortest (65.6 cm) and thinnest tubers (3.9 cm) with the lowest fresh weight (953.4 g) and yield (28 532.8 kg/hm2) among the 14 fertilizer combinations. The experimental data could be fitted to single-variable quadratic and binary quadratic models but not to a ternary quadratic polynomial model. Appropriate N, P, and K fertilizer application rates increased Chinese yam yield. However, excessive fertilization lowered the yield. Chinese yam yield was significantly and strongly correlated with the amounts of N, P, and K fertilizer applied.
CONCLUSION
Based on the single variable quadratic and binary quadratic models, we propose that the quantities of N, P, and K fertilizer used to grow 1 hm2 'Qinfeng' Chinese yam should be 360-388.3, 90-100.95, and 416.3-675 kg, respectively.
3.Effect of down-regulation of let-7c/g on triggering a double-negative feedback loop and promoting restenosis.
Qian ZHANG ; Xiaojun ZHOU ; Xianzhi LI ; Shuai YAO ; Shan JIANG ; Rui ZHANG ; Zhiwei ZOU ; Lin LIAO ; Jianjun DONG
Chinese Medical Journal 2023;136(20):2484-2495
BACKGROUND:
Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) are the main causes of restenosis (RS) in diabetic lower extremity arterial disease (LEAD). However, the relevant pathogenic mechanisms are poorly understood.
METHODS:
In this study, we introduced a "two-step injury protocol" rat RS model, which started with the induction of atherosclerosis (AS) and was followed by percutaneous transluminal angioplasty (PTA). Hematoxylin-eosin (HE) staining and immunohistochemistry staining were used to verify the form of RS. Two-step transfection was performed, with the first transfection of Lin28a followed by a second transfection of let-7c and let-7g, to explore the possible mechanism by which Lin28a exerted effects. 5-ethynyl-2΄-deoxyuridine (EdU) and Transwell assay were performed to evaluate the ability of proliferation and migration of VSMCs. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect the expression of Lin28a protein and let-7 family members.
RESULTS:
Using a combination of in vitro and in vivo experiments, we discovered that let-7c, let-7g, and microRNA98 (miR98) were downstream targets of Lin28a. More importantly, decreased expression of let-7c/let-7g increased Lin28a, leading to further inhibition of let-7c/let-7g. We also found an increased level of let-7d in the RS pathological condition, suggesting that it may function as a protective regulator of the Lin28a/let-7 loop by inhibiting the proliferation and migration of VSMCs.
CONCLUSION
These findings indicated the presence of a double-negative feedback loop consisting of Lin28a and let-7c/let-7g, which may be responsible for the vicious behavior of VSMCs in RS.
Rats
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Animals
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Down-Regulation
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MicroRNAs/metabolism*
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Feedback
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Cell Proliferation/genetics*
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Atherosclerosis
4.Effect of preoperative peripheral blood neutrophil to lymphocyte ratio and platelet distribution width on clinical prognosis of childhood medulloblastoma
Jinqiao ZHOU ; Yufeng LIU ; Ke LI ; Xianzhi LIU ; Zhenyu ZHANG
Chinese Journal of Applied Clinical Pediatrics 2020;35(10):797-800
Objective:To investigate the effect of peripheral blood neutrophil to lymphocyte ratio (NLR) and platelet distribution width (PDW) on the clinical prognosis of childhood medulloblastomaMethods:Clinical data and survival data of 76 pediatric patients who were diagnosed as medulloblastoma by post-operative pathology in the First Affiliated Hospital of Zhengzhou University from January 2009 to December 2016 were collected. Kaplan- Meier method was used to calculate the overall survival(OS) and progression free survival(EFS) rates, Log- rank test was employed to compare the survival rates of different groups, and Cox proportional hazards regression model was used for multivariate analysis. Results:The log- rank test revealed that 5-year PFS rate and OS rate (22.2%, 22.2%) in the high NLR group (NLR>4.94) were significantly lower than those in the low NLR group (NLR≤4.94) (45.6%, 55.7%), and the differences were statistically significant(PFS: P=0.009, OS: P=0.001), and the 5-year PFS and OS (52.3%, 66.5%) of the high PDW group (PDW>15.90) were significantly higher than those in the low PDW group (PDW ≤ 15.90) (27.1%, 32.5%), and the differences were statistically significant(PFS: P=0.032, OS: P=0.039). Univa-riate analysis showed that the extent of resection (PFS: P=0.006, OS: P=0.009), and postoperative radiotherapy (PFS: P=0.011, OS: P=0.001) and postoperative radiotherapy(PFS: P=0.011, OS: P=0.001) were the factors influencing the prognosis of children with medulloblastoma.Multivariate Cox proportional hazards regression model analysis suggested that no postoperative radiotherapy (PFS: P=0.048, OS: P=0.008), NLR>4.94 (PFS: P=0.023, OS: P=0.003) and PDW≤15.90 (PFS: P=0.028, OS: P=0.006) were the independent risk factors for the prognosis of childhood medulloblastoma. Conclusions:Increased NLR and decreased PDW indicate unfavorable prognosis of the childhood medulloblastoma.Therefore, preoperative NLR and PDW may be the potential prognostic markers for childhood medulloblastoma.
5.Pseudolaric Acid B Inhibits Proliferation, Invasion and Epithelial-to-Mesenchymal Transition in Human Pancreatic Cancer Cell
Xiaoyu LI ; Xianzhi ZHAO ; Wen SONG ; Zibin TIAN ; Lin YANG ; Qinghui NIU ; Qi ZHANG ; Man XIE ; Bin ZHOU ; Yonghong XU ; Jun WU ; Cuiping ZHANG
Yonsei Medical Journal 2018;59(1):20-27
PURPOSE: This study was aimed to investigate the effect of pseudolaric acid B (PAB) on proliferation, invasion and epithelial-to-mesenchymal transition (EMT) in pancreatic cancer cells and to explore the possible mechanism. MATERIALS AND METHODS: The pancreatic cancer cell line SW1990 was cultured and treated with PAB dose- and time-dependent manners. Cell proliferation and invasion ability were measured by MTT assay and Matrigel/Transwell test, respectively. Semi-quantitative real-time polymerase chain reaction and Western blotting were conducted to detect the expression of EMT markers and the key molecules. Finally, nude mice subcutaneous transplantation tumor model was used to confirm the therapy efficacy of PAB. RESULTS: PAB could inhibit SW1990 cell proliferation and invasion in time- and dose-dependent manners. Vimentin, fibronectin, N-cadherin, Snail, Slug, YAP, TEAD1, and Survivin were down-regulated (p < 0.01), while E-cadherin, caspase-9, MST1, and pYAP were up-regulated (p < 0.05). Combined PAB and gemcitabine treatment markedly restricted the tumor growth compared with gencitabin or PAB alone groups. CONCLUSION: PAB could inhibit the proliferation and invasion ability of pancreatic cancer cells through activating Hippo-YAP pathway and inhibiting the process of EMT.
Animals
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Antineoplastic Agents/pharmacology
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Antineoplastic Agents/therapeutic use
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Biomarkers, Tumor/metabolism
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Cadherins
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Cell Line, Tumor
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Cell Movement
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Cell Proliferation/drug effects
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Cytokines
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Deoxycytidine/analogs & derivatives
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Deoxycytidine/pharmacology
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Deoxycytidine/therapeutic use
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Diterpenes/pharmacology
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Diterpenes/therapeutic use
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Epithelial-Mesenchymal Transition/drug effects
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Female
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Humans
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Mice, Nude
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Neoplasm Invasiveness
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Pancreatic Neoplasms/diet therapy
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Pancreatic Neoplasms/pathology
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Real-Time Polymerase Chain Reaction
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Signal Transduction/drug effects
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Vimentin/metabolism
6.Evaluation of performance between ABI 7500 and TL 988 fluorescent re-al-time PCR instrument
Qianglong PAN ; Chenglin ZHOU ; Xianzhi YANG ; Guoliang XIE ; Dawei CUI
China Modern Doctor 2015;(2):155-156,160
Objective To explore the differences between ABI 7500 and TL 988 fluorescent quantitative instrument for polymerase chain reaction. Methods Based on EP9-A2 Files, quantitative influenza A virus positive controls of five kinds of concentrations were detected by two kinds of fluorescence quantitative PCR instruments at the same time, and each sample was detected 20 times. The precision and sensitivity between the two kinds of instruments were compared. Results Inner group CV of TL 988 ranged from 0.68% to 1.52%. The CV for ABI 7500 ranged from 1.93% to 2.69%. The high (107, 106, 105 copies/mL) levels of IFVA positive control were detected by ABI 7500 and TL 988 instruments, and there were no significant difference between the results(P>0.05) while the low(104, 103 copies /mL)levels were de-tected by ABI 7500 and TL 988 instruments, and there was a significant difference between the results(P<0.05). When samples were detected at the level of 103 copies/mL, the positive detection rate was 100% with TL 988 instrument, but the positive detection rate was 0 with ABI 7500 instrument. The related index of ABI 7500 and TL 988 was better (r=0.998). Conclusion The performances in precision and sensitivity of TL 988 PCR instrument are better than ABI 7500 PCR.
7.Peripheral blood stem cell transplantation for treating nervous system disease: Possibility, feasibility, and confidence
Shaoyu ZHOU ; Chengwen ZHANG ; Bijia TANG ; Xianzhi ZENG
Chinese Journal of Tissue Engineering Research 2010;14(1):142-146
BACKGROUND: Peripheral blood stem cells (PBSCs) can be enhanced and differentiates into all kinds of neural cells and produce several kinds of neural growth factors in vitro, and have been used to treat several kinds of neural diseases and they showed satisfactory outcomes. It is now a research focus to optimize the culture conditions to induce the stem cell to differentiate into neural cells. On the other hand, it also becomes a focus of its amplification and differentiation. OBJECTIVE: To make a review on researches on treating brain lesion, genetic defection or degenerating diseases using PBSCs during recent years.METHODS: Researching Pubmed database(1999-01/2009-09), using "peripheral blood stem cell, neural, repair" as research words and these Chinese words in CNKI database(1999-01/2009-09). RESULTS AND CONCLUSION: 221 articles were collected altogether including 41 Chinese articles and 180 English articles. Totally 27 articles were adapted altogether after rejecting published early, repeat and similar articles. PBSCs can be enhanced and differentiates into all kinds of neural cells in vitro, and delivered into central neural system and improve the function of lesion neural area. It is not fully understood the differentiation mechanism of peripheral blood stem cells, but numerous studies have shown that surface marker antigen of peripheral blood stem cells is closely associated with its biological characteristics. There are many studies addressing surface marker of peripheral blood stem cells, but researchers cannot identify a reliable marker that can directly label peripheral blood stem cells. Function of CD34 remains unclear, so the principle of CD34 as a marker of hematopoietic stem cells receives challenge. It is necessary to do further researches on issues on how to enhance survival rate of PBSCs in vivo and the tendency to differentiate into neural cells.
8.Analysis on "Heliang & Heli" Fund Award in military health system
Chinese Journal of Medical Science Research Management 2010;23(2):113-115
By analyzing the Military Health System winning "Heliang & Heli" Fund Award between 1994 and 2009,we found that the Military Health System possessed a good scientific basis and high level,but with an unbalanced scientific research development,which suggests the Military Health System should further strengthen its independent innovation ability.
9.Effects and adverse drug reactions of mtrisone in the treatment of patients with severe acute respiratory syndrome
Rui WANG ; Xiaoqing ZHOU ; Jun DONG ; Rong WEI ; Xiutang CAO ; Yachun ZHOU ; Jin WANG ; Daihong GUO ; Kun CHEN ; Jian ZHOU ; Jiesong WANG ; Xiumei ZHU ; Beibei LIANG ; Yanping XU ; Xianzhi ZHOU
Chinese Journal of Clinical Pharmacology and Therapeutics 2004;0(09):-
AIM: To study effects and adverse drug reactions of mtrisone in the treatment of patients with severe acute respiratory syndrome. METHODS: The information of the medications in 680 patients with severe acute respiratory syndrome (SARS) in Xiaotangshan Hospital was collected by HIS system and the effects and ADRs of metrisone were staiated. RESULTS: The kinds of drugs of SARS patients who had been cured by metrisone were more than those which were not cured by metrisone. Condition of SARS patients who had been cured by metrisone was more serious than those which were not cured by metrisone. The ADRs rate, blood glucose and leukocyte of SARS patients who had been cured by metrisone are higher than those which were not cured by metrisone while blood K+ is lower. CONCLUSION: The utilization of metrison to SRAS patient should be more cautious to balance the effects and ADRs of metrisone.
10.Construction of recombinant adenovirus vector Ad-elafin for nature antibiotic elafin gene and its expression in primary airway epithelium
Journal of Third Military Medical University 2003;0(11):-
Objective To construct the recombinant adenovirus for nature antibiotic elafin protein and observe its expression in primary airway epithelia.Methods The elafin gene was amplified by RT-PCR from lung tissues,then the fragment was inserted into the multiple clone site of pAdTrack-CMV.The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183.The candidate clone was further analyzed by restriction endonuclease digestion,PCR,and sequencing.Then the recombinant adenovirus plasmid was digested with PacⅠ and transfected into 293 cells for packaging and amplification.Infection titer and rate were monitored by green fluorescent protein(GFP)expression.Finally the primary airway epithelium was infected with Ad-elafin,and the GFP expression was observed by fluorescence microscope in epithelial cells 24 h after transfection.The elafin protein was detected by ELISA in the supernatant.Results Restriction endonuclease and PCR confirmed that elafin gene was cloned into the adenovirus vector successfully.Twenty-four hours after transfection,the GFP expression was seen by fluorescence microscopy.The concentration of elafin protein was(2.2?0.4)ng/ml in supernatant of transfected group,while that of control group was(1.9?0.3)ng/ml,P

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