1.Drug resistance induction and analysis of differential expression protein on adult Schistosoma j aponicum induced by ED50 PZQ
Lanlan DONG ; Jing XU ; Bo ZHAO ; Song LIANG ; Yanyan WANG ; Zhixun GUAN ; Yun CAO ; Chaoming XIA
Chinese Journal of Zoonoses 2014;(12):1171-1180
ABSTRACT:In the present study ,we aimed to identify differentially expressed proteins between induced worms (the infec‐ted mice were treated intragastrically with ED50 PZQ) and uninduced worms (control group) for clarifying the mechanism of PZQ .ED50 PZQ was used to administrate mice that were infected with S .japonicum via intragastric incubation for consecutive‐ly 30 days .Twenty‐one days later ,mice were sacrificed after treatment with 200 mg/kg PZQ for continuously five days ,and the male worms were obtained and some of them were subjected in DMEM medium with different concentrations of PZQ in vitro for 16 hours .Then the worms were washed twice and incubated in PZQ‐free medium for 72 hours .Compared with control group ,the induced worms had lesser sensitivity to PZQ .The survival rate of induced worms was 75 .6% in vitro when the con‐centration of PZQ was 112 mol/L (the concentration was 8 times of uninduced worms Lethal Concentration ) ,significantly higher than that in the uninduced worms (11 .1% ,P<0 .05) ,showing obviously tolerance .The other induced and uninduced worms were acquired and collected for 2D‐DIGE and MALDI‐TOF‐MS ,and combined with bioinformatics to analyse the func‐tion of the identified protein .Thirty differential expression proteins were confirmed between induced and uninduced worms ,in‐cluding 12 proteins up‐regulated and 18 proteins down‐regulated .These proteins respectively ascribed to cytoskeleton‐associat‐ed protein ,glucose and energy metabolism enzymes ,stress proteins ,thioredoxin peroxidase enzymes ,and other protease .Up‐or down‐regulation of these differential proteins indicated that PZQ promote or inhibit the expression of some specific genes . These findings may help to clarify the mechanism of PZQ ,simultaneously ,providing a scientific basis for exploring new vaccine candidate antigens and targets for drug therapy .
2.Electrospun materials applied in drug delivery system
Zhifeng ZHAO ; Zhixun YANG ; Feng LI ; Hamushan MUSHA ; Yifan SHI ; Jian FEI ; Jun ZHANG ; Wenguo CUI
Chinese Journal of Tissue Engineering Research 2017;21(2):303-308
BACKGROUND:Recently, electrospun materials have been extensively applied in the drug delivery system. OBJECTIVE:To overview the application prospect of electrospun materials in drug delivery systems. METHODS:A computer-based search of PubMed and NCBI databases was performed for literatures about the research progress of electrospinning in tissue engineering and chemotherapy published within the past 10 years using the keywords of“electrospinning, drug delivery system, nanofibers, electrospun materials”.RESULTS AND CONCLUSION:Compared with traditional materials, electrospun stents hold good versatility and control able parameters, thus granting its unique advantage under various physiological conditions. Current drug-loaded materials composed of natural products, synthetic polymers and blended materials;as to drugs, there are antibiotics, chemotherapy medication, DNA and protein. Electrospun materials have been used in tissue engineering, cancer chemotherapy and wound healing. We focus on not only the application progress of electrospun materials in traditional treatments, but also its usage, condition-control ed drug release and living-cel carrying. Electrospun materials combined with various drug-loaded present a broad prospect.
3.Progression of diagnosis and treatment in primary malignant small bowel tumor.
Zhixun ZHAO ; Xu GUAN ; Yinggang CHEN ; Xishan WANG
Chinese Journal of Gastrointestinal Surgery 2017;20(1):117-120
Primary malignant small bowel tumor as a rare kind of intestinal tumor is associated with a poor prognosis. The pathological types were various and complicated, such as adenocarcinoma, neuroendocrine tumor, malignant lymphoma, and malignant stromal tumor. The atypical early stage symptom resulted in difficult diagnosis at early stage, high misdiagnosis rate and lack of standard therapy schemes and means. In the past, X-ray, CT, MRI, and PET-CT were the main examination methods for primary small bowel tumor. However, with the development of radiology, a series of new diagnosis methods, including electronic enteroscopy, capsule endoscopy, multi-slice spiral CT enteroclysis and so on, promotes the diagnosis accurate rate. Surgery is still the most important method in the small bowel tumor treatment, and the alternative of the surgical method should depend on the tumor location, size and relationship with the adjacent organs. Application of the laparoscopic surgery for the small bowel tumor is still in the initial stage. Besides, some researches have confirmed that chemotherapy, radiotherapy, target therapy and endocrinotherapy have effects on the specific kind of small bowel tumor. Therefore this article will review the epidemiology, pathology, diagnosis and treatment of the primary malignant small bowel tumors.
Adenocarcinoma
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diagnosis
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epidemiology
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pathology
;
therapy
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Capsule Endoscopy
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Gastrointestinal Stromal Tumors
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diagnosis
;
epidemiology
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pathology
;
therapy
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Humans
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Intestinal Neoplasms
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diagnosis
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epidemiology
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pathology
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therapy
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Intestine, Small
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diagnostic imaging
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pathology
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Laparoscopy
;
methods
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Lymphoma
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diagnosis
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epidemiology
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pathology
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therapy
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Neuroendocrine Tumors
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diagnosis
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epidemiology
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pathology
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therapy
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Tomography, Spiral Computed
4.The clinical significance of lateral pelvic sentinel lymph node biopsy using indocyanine green fluorescence navigation in laparoscopic lateral pelvic lymph node dissection
Hao SU ; Zheng XU ; Mandula BAO ; Shou LUO ; Jianwei LIANG ; Wei PEI ; Xu GUAN ; Zheng LIU ; Zheng JIANG ; Mingguang ZHANG ; Zhixun ZHAO ; Weisen JIN ; Haitao ZHOU
Chinese Journal of Oncology 2024;46(2):140-145
Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.
5.The clinical significance of lateral pelvic sentinel lymph node biopsy using indocyanine green fluorescence navigation in laparoscopic lateral pelvic lymph node dissection
Hao SU ; Zheng XU ; Mandula BAO ; Shou LUO ; Jianwei LIANG ; Wei PEI ; Xu GUAN ; Zheng LIU ; Zheng JIANG ; Mingguang ZHANG ; Zhixun ZHAO ; Weisen JIN ; Haitao ZHOU
Chinese Journal of Oncology 2024;46(2):140-145
Objectives:This study aims to explore the clinical significance of lateral pelvic sentinel lymph node biopsy (SLNB) using indocyanine green (ICG) fluorescence navigation in laparoscopic lateral pelvic lymph node dissection (LLND) and evaluate the accuracy and feasibility of this technique to predict the status of lateral pelvic lymph nodes (LPLNs).Methods:The clinical and pathological characteristics, surgical outcomes, lymph node findings and perioperative complications of 16 rectal cancer patients who underwent SLNB using ICG fluorescence navigation in laparoscopic LLND in the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College during April 2017 and October 2022 were retrospectively collected and analyzed. The patients did not receive preoperative neoadjuvant radiotherapy and presented with LPLNs but without LPLN enlargement (MRI showed the maximum short axes of the LPLNs were ≥5 mm and <10 mm at first visit).Results:All 16 patients were successfully performed SLNB using ICG fluorescence navigation in laparoscopic LLND. Three patients underwent bilateral LLND and 13 patients underwent unilateral LLND. The lateral pelvic sentinel lymph nodes (SLNs) were clearly fluorescent before dissection in 14 patients and the detection rate of SLNs for these patients was 87.5%. Lateral pelvic SLN metastasis was diagnosed in 2 patients and negative results were found in 12 patients by frozen pathological examinations. Among the 14 patients in whom lateral pelvic SLNs were detected, the dissected lateral pelvic non-SLNs were all negative. All dissected LPLNs were negative in two patients without fluorescent lateral pelvic SLNs. The specificity, sensitivity, negative predictive value, and accuracy was 85.7%, 100%, 100%, and 100%, respectively.Conclusions:This study indicates that lateral pelvic SLNB using ICG fluorescence navigation shows promise as a safe and feasible procedure with good accuracy. This technique may replace preventive LLND for locally advanced lower rectal cancer.
6.Expression and refolding of OLA Ⅰ protein with peptides derived from sheeppox virus.
Zhanhong WANG ; Zhixun ZHAO ; Guohua WU ; Yang DENG ; Guoqiang ZHU ; Fangyan ZHAO ; Zengjun LU ; Qiang ZHANG
Chinese Journal of Biotechnology 2022;38(1):139-147
The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals
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Capripoxvirus
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Epitopes, T-Lymphocyte/genetics*
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Peptides/genetics*
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Poxviridae Infections
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Sheep
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Sheep Diseases
7.A method for immortalizing swine monoclonal B cells secreting anti-PRRSV antibodies.
Jian WANG ; Jing ZHANG ; Kun LI ; Pu SUN ; Guoxiu LI ; Jiaoyang LI ; Yimei CAO ; Zhixun ZHAO ; Hong YUAN ; Yuanfang FU ; Pinghua LI ; Dong LI ; Zaixin LIU ; Zengjun LU
Chinese Journal of Biotechnology 2022;38(8):2872-2882
Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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Antibodies, Viral
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Humans
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Porcine Reproductive and Respiratory Syndrome/prevention & control*
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Porcine respiratory and reproductive syndrome virus/genetics*
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Swine