1.Quantitative analysis of cell tracing by in vivo imaging system.
Junmeng, ZHENG ; Lijun, XU ; Hongmin, ZHOU ; Weina, ZHANG ; Zhonghua, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):541-5
In vivo imaging system (IVIS) is a new and rapidly expanding technology, which has a wide range of applications in life science such as cell tracing. By counting the number of photons emitted from a specimen, IVIS can quantify biological events such as tumor growth. We used B16F10-luc-G5 tumor cells and 20 Babl/C mice injected subcutaneously with B16F10-luc-G5 tumor cells (1x10(6) in 100 muL) to develop a method to quantitatively analyze cells traced by IVIS in vitro and in vivo, respectively. The results showed a strong correlation between the number of tumor cells and the intensity of bioluminescence signal (R (2)=0.99) under different exposure conditions in in vitro assay. The results derived from the in vivo experiments showed that tumor luminescence was observed in all mice by IVIS at all days, and there was significant difference (P<0.01) between every two days from day 3 to day 14. Moreover, tumor dynamic morphology could be monitored by IVIS when it was invisible. There was a strong correlation between tumor volume and bioluminescence signal (R (2)=0.97) by IVIS. In summary, we demonstrated a way to accurately carry out the quantitative analysis of cells using IVIS both in vitro and in vivo. The data indicate that IVIS can be used as an effective and quantitative method for cell tracing both in vitro and in vivo.
2.Repairing atrial septal defects with bilateral lung transplantation in treatment of ASD with Eisenmenger's syndrome
Junmeng ZHENG ; Jinxiong ZHEN ; Baoping DENG ; Weizhao HUANG ; Hongyu YE ; Yi LIANG ; Haiming JIANG
Chinese Journal of Organ Transplantation 2016;37(11):668-671
Objective To investigate the feasibility and the perioperative treatments of repairing atrial septal defects and bilateral lung transplantation in treatment of ASD with Eisenmenger's syndrome.Methods On May 24th,2016,one case of atrial septal defect with Eisenmenger's syndrome underwent repair for the atrial septal defect with bilateral sequential lung transplantation aided by extracorporeal circulation ECMO in Zhongshan People's Hospital.In strict accordance to the procedure,the lung was collected and preserved with the HTK solution.The operation went smoothly,the cold ischemic time for the right lung was 120 min and that for the left lung was 260 min.Postoperatively,Tacrolimus (FK506) + Mycophenolate Mofetil (MMF) + adrenal cortical hormone Sanlian immunosuppression was used.Results At 60th h post-operation the patient was hemodynamically stable,with good oxygenation after removal of the ECMO.At 7th day post-operation the patient was extubated.At 12th-20th day post-operation bacterial culture of the sputum was found to be positive and the patient was treated with the corresponding sensitive antibiotics and recovered.Cardiac ultrasound before discharge showed that there was no residual shunt.One month after discharge the cardiopulmonary function of the patient was quite good,chest X-ray showed that the bilateral transplanted lungs were clear and the patient's quality of life was much better.Conclusion Cardiac malformation repairs with bilateral lung transplantation is a feasible treatment for patients with end-stage Eisenmenger's syndrome with bi-directional shunts.The key to success is sufficient preoperative preparation,and a smooth operation with the timely and effective treatment of postoperative complications.
3.Clinical application value of serum exosomal circRPS6 in the diagnosis and prognosis assessment of patients with colorectal cancer
Liuchang ZHENG ; Junmeng LI ; Peiming ZHENG
Chinese Journal of Laboratory Medicine 2024;47(3):272-277
Objective:To investigate the expression and clinical application value of exosomal circRPS6 in serum of colorectal cancer (CRC) patients.Methods:Peripheral serum samples were collected from 115 CRC patients admitted to Henan Provincial People′s Hospital from January 2019 to December 2020. There were 68 males and 47 females, aged (63.0±9.5) years. Meanwhile, one hundred and twenty healthy subjects from the same period wereenrolled, with 70 males and 50 females, aged (61.0±10.7) years. In addition, sixty pairs of tumor and adjacent tissue specimens from CRC patients undergoing surgical treatment were collected. The circRPS6 expression in serum exosome and tissue of CRC patients were detected via real-time fluorescence quantitative PCR (RT-qPCR), and its relationship with clinicopathological features and prognosis of CRC patients were also investigated. The levels of CEA and CA19-9 in serum were detected by electrochemiluminescence assay. The ROC curve and AUC were used to estimate the diagnostic capacity. Univariate and multivariate regression analysis was performed using Cox proportional hazard analysis.Results:The expression level of circRPS6 in CRC tissue was significantly higher than that in adjacent tissue( Z=5.38, P<0.001). Compared with healthy control, the expression of serum exosomal circRPS6 was significantly upregulated in the CRC group( t=14.52, P<0.001). ROC curve analysis results showed that the AUC of exosomal circRPS6 was 0.882, which had a higher diagnostic efficacy in CRC patients than CEA and CA19-9 detection. There was a positive correlation between the expression level of exosomal circRPS6 with TNM stage and lymph node metastasis and distant metastasis( P<0.05). Kaplan-Meier survival analysis revealed that CRC patients with low exosomal circRPS6 levels had a much longer average survival time compared with those in high group. Moreover,multivariate analysis results indicated that exosomal circRPS6 was an independent prognostic factor in colorectal cancer. Conclusion:Exosomal circRPS6 is highly expressed in the serum of CRC patients and correlated with malignant progression and poor prognosis, which is expected to be a potential marker for the diagnosis and prognosis evaluation of CRC patients.
4.A novel bone marrow transplantation strategy for donor-specific tolerance induction after heart transplantation
Kequan GUO ; Xu MENG ; Yuanlong YU ; Jie HAN ; Haiming JIANG ; Xiaojun XU ; Xiaojun LU ; Yixin JIA ; Junmeng ZHENG ; Haibo ZHANG ; Yan LI ; Tie ZHENG ; Chunlei XU ; Wen ZENG ; Jiangang WANG ; Yongqiang CUI ; Tiange LUO ; Jun WANG ; Susumu IKEHARA
Chinese Journal of Organ Transplantation 2011;32(1):32-35
Objective To investigate a new strategy of bone marrow transplantation (BMT) for donor-specific tolerance induction after heart transplantation. Methods Donor bone marrow cells (BMCs)were harvested simultaneously with donor cardiac graft using modified perfusion method (PM) ,then stored in a -80 ℃ refrigerator after filtration and centrifugation. Whole BMCs (IBM-BMT) (monocytes 1.2 ×107/kg,CD34+ cells 2.38× 105/kg) in host iliac bones were injected into the bone marrow cavity 40 days after heart transplantation. Preconditoning regimens that consisted of fludarabine, antithymoctye globin and total lymphoid irradiation were performed 3 days before BMT. Tacrolimus (Tac) was administrated intravenously after BMT or orally in conjunction with mycophenolate mofetil (MMF) 3 weeks later.Cyclosporine and MMF were orally administrated 6 weeks later. Donor chimerism was detected using short tandem repeats-polymerase chain reaction in monocytes from peripheral blood at the 2nd,4th, 8th or 12th week after BMT or BMCs at the 4th, 8th or 12th week after BMT. Intramyocardium electrocardiography examination or endomyocardial biopsy was performed weekly or monthly respectively. Mixed lymphocyte reactions (MLR) were performed 3 months after BMT. Results Donor chimerism in monocytes in peripheral blood or BMCs in iliac bones measured at the 1 st,2nd and 3rd month after BMT was 26.3%, 19.1%,4.8% ,and 46.3%, 24.4%, 7.6%, respectively. After 3-month follow-up, there was no rejection confirmed by endomyocardial biopsy or intramyocardium electrocardiography. Echocardiography revealed that the diastolic and systolic function of the cardiac graft was maintained well 3 months after BMT. MLR revealed donor-specific hyporesponsiveness while immunocompetence was preserved to third-party antigens. Conclusion These findings indicate that the two-stage BMT strategy is a safe and feasible method for the induction of donor-specific tolerance via stable mixed chimerism and needs to be further confirmed after a long-term observation.
5.Quantitative Analysis of Cell Tracing by in vivo Imaging System
ZHENG JUNMENG ; XU LIJUN ; ZHOU HONGMIN ; ZHANG WEINA ; CHEN ZHONGHUA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):541-545
In vivo imaging system(IVIS)is a new and rapidly expanding technology,which has a wide range of applications in life science such as cell tracing.By counting the number of photons emitted from a specimen,IVIS can quantify biological events such as tumor growth.We used B16F10-luc-G5 tumor cells and 20 Babl/C mice injected subcutaneously with B16F10-luc-G5 tumor cells(1×106 in 100 μL)to develop a method to quantitatively analyze cells traced by IVIS in vitro and in vivo,respectively.The results showed a strong correlation between the number of tumor cells and the intensity of bioluminescence signal(R2=0.99)under different exposure conditions in in vitro assay.The results derived from the in vivo experiments showed that tumor luminescence was observed in all mice by IVIS at all days,and there was significant difference(P<0.01)between every two days from day 3 to day 14.Moreover,tumor dynamic morphology could be monitored by IVIS when it was invisible.There was a strong correlation between tumor volume and bioluminescence signal(R2=0.97)by IVIS.In summary,we demonstrated a way to accurately carry out the quantitative analysis of cells using IVIS both in vitro and in vivo.The data indicate that IVIS can be used as an effective and quantitative method for cell tracing both in vitro and in vivo.
6.Application and research progress of nanotechnology in atherosclerosis
Tingting WANG ; Lili YU ; Xiangli SHEN ; Junmeng ZHENG ; Yushan CHEN ; Shasha SHANG ; Jianru WANG
The Journal of Practical Medicine 2024;40(1):53-58
Atherosclerosis(AS)is a common cardiovascular disease,and its treatment and prevention have been the focus of medical research.AS an emerging technology,nanotechnology has unique advantages and plays an important role in the prevention,diagnosis and treatment of AS.This paper reviews the latest research on the application of nanotechnology in AS diseases,systematically discusses the role of nanotechnology in the diag-nosis and treatment of AS,and comprehensively analyzes the effects of nano-drug carriers based on different sur-face trimmers,loading diagnostic and therapeutic drugs so as to monitordisease progression of AS and its targeted treatment.The aim is to provide new thought for the clinical treatment of AS.
7.Ablation of Akt2 and AMPKα2 rescues high fat diet-induced obesity and hepatic steatosis through Parkin-mediated mitophagy
Shuyi WANG ; Jun TAO ; Huaguo CHEN ; Machender R KANDADI ; Mingming SUN ; Haixia XU ; Gary D LOPASCHUK ; Yan LU ; Junmeng ZHENG ; Hu PENG ; Jun REN
Acta Pharmaceutica Sinica B 2021;11(11):3508-3526
Given the opposing effects of Akt and AMP-activated protein kinase (AMPK) on metabolic homeostasis, this study examined the effects of deletion of Akt2 and AMPKα2 on fat diet-induced hepatic steatosis. Akt2–Ampkα2 double knockout (DKO) mice were placed on high fat diet for 5 months. Glucose metabolism, energy homeostasis, cardiac function, lipid accumulation, and hepatic steatosis were examined. DKO mice were lean without anthropometric defects. High fat intake led to adiposity and decreased respiratory exchange ratio (RER) in wild-type (WT) mice, which were ablated in DKO but not Akt2−/− and Ampkα2−/− mice. High fat intake increased blood and hepatic triglycerides and cholesterol, promoted hepatic steatosis and injury in WT mice. These effects were eliminated in DKO but not Akt2−/− and Ampkα2−/− mice. Fat diet promoted fat accumulation, and enlarged adipocyte size, the effect was negated in DKO mice. Fat intake elevated fatty acid synthase (FAS), carbohydrate-responsive element-binding protein (CHREBP), sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), peroxisome proliferator-activated receptor-α (PPARα), PPARγ, stearoyl-CoA desaturase 1 (SCD-1), phosphoenolpyruvate carboxykinase (PEPCK), glucose 6-phosphatase (G6Pase), and diglyceride O-acyltransferase 1 (DGAT1), the effect was absent in DKO but not Akt2−/− and Ampkα2−/− mice. Fat diet dampened mitophagy, promoted inflammation and phosphorylation of forkhead box protein O1 (FoxO1) and AMPKα1 (Ser485), the effects were eradicated by DKO. Deletion of Parkin effectively nullified DKO-induced metabolic benefits against high fat intake. Liver samples from obese humans displayed lowered microtubule-associated proteins 1A/1B light chain 3B (LC3B), Pink1, Parkin, as well as enhanced phosphorylation of Akt, AMPK (Ser485), and FoxO1, which were consolidated by RNA sequencing (RNAseq) and mass spectrometry analyses from rodent and human livers. These data suggest that concurrent deletion of Akt2 and AMPKα2 offers resilience to fat diet-induced obesity and hepatic steatosis, possibly through preservation of Parkin-mediated mitophagy and lipid metabolism.
8.Erratum: Author correction to 'Ablation of Akt2 and AMPKα2 rescues high fat diet-induced obesity and hepatic steatosis through Parkin-mediated mitophagy' Acta Pharmaceutica Sinica B 11 (2021) 3508-3526.
Shuyi WANG ; Jun TAO ; Huaguo CHEN ; Machender R KANDADI ; Mingming SUN ; Haixia XU ; Gary D LOPASCHUK ; Yan LU ; Junmeng ZHENG ; Hu PENG ; Jun REN
Acta Pharmaceutica Sinica B 2023;13(2):897-898
[This corrects the article DOI: 10.1016/j.apsb.2021.07.006.].