1.Growth and secretion features of microencapsulated human adrenal pheochromocytoma cells in artificial cerebrospinal fluid
Xiao LI ; Xinsheng ZHANG ; Haijie LUO ; Shuo LUAN ; Qing WAN ; Shaoling WU ; Chao MA
Chinese Journal of Tissue Engineering Research 2015;(38):6114-6120
BACKGROUND:Microencapsulated cels are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cels is unclear.
OBJECTIVE:To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cels in artificial cerebrospinal fluid.
METHODS: The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cels that were then cultured in artificial cerebrospinal fluid. Primary cels were covered with alginate-polylysine-alginate microcapsules, and then the cel morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cel culture medium were detected by enzyme-labeled immunosorbent assay (ELISA). We used cel counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cels in artificial cerebrospinal fluid.
RESULTS AND CONCLUSION:Microcapsulated human pheochromocytoma cels were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cels, the proliferation rate of microcapsulated cels increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cels compared to the non-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cels can survive wel and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cels from different tumor tissues have stable secretory function.
2.Exercise effects on pain relief and extracellular matrix production in a rat model of intervertebral disc degeneration
Haijie LUO ; Songjian KE ; Caina LIN ; Qing WAN ; Xiao LI ; Cuicui LIU ; Chao MA ; Shaoling WU
Chinese Journal of Tissue Engineering Research 2017;21(20):3176-3182
BACKGROUND: Exercise has been proved to accelerate the proliferation of intervertebral disc cells and extracellular matrix production in healthy rats. For the degenerative intervertebral disc, whether exercise also has positive effects on its cell proliferation, extracellular matrix production or pain relief remains unclear. OBJECTIVE: To investigate the effect of exercise on the extracellular matrix production in a rat model of intervertebral disc degeneration.METHODS: A rat model of intervertebral disc degeneration was prepared by Freund's complete adjuvant injection into the intervertebral disc at L5-6 levels. Then, the model rats were allowed to have a rest for 2 weeks. All rats were then randomly divided into exercise and control groups. Rats in the exercise group were forced to run every day, while the controls allowed free activities in the cage. The behavioral tests were performed at 7, 14, 28, 42, 56 and 70 days after modeling; meanwhile, the intervertebral disc samples were collected used for alcian blue staining and immunohistochemical staining to detect the levels of proteoglycan, aggrecan and collagen type Ⅱ in the intervertebral disc cells, respectively.RESULTS AND CONCLUSION: Vocalization threshold on the rat back of punctured disc was significantly decreased, while grooming and wet-dog shaking were significantly increased at 7 days after modeling compared with the baseline (P < 0.05), suggesting that Freund's complete adjuvant injection successfully induces disc degeneration, hyperalgesia and abnormal behaviors. Further, the vocalization threshold and wet-dog shaking in the exercise group showed significant improvement compared with the control group after 14 days of exercise (P < 0.05), while the grooming was significantly reduced until the 28th day (P < 0.01), indicating that exercise can alleviate pain caused by disc degeneration in model rats. At 21 days after modeling, the levels of proteoglycan, aggrecan and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus were significantly decreased compared with the baseline (P < 0.01), indicating the occurrence of disc degeneration. After 14 days of training, the levels of proteoglycan, aggrecan, and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus in the exercise group were significantly increased compared with the control group (P < 0.01). Moreover, after 8-week exercise, the level of proteoglycan in the nucleus pulposus and annulus fibrosus in the exercise group was increased by 4-5 times compared with the control group, and levels of aggrecan and collagen type Ⅱ in the nucleus pulposus in the exercise group also was increased by 3-4 times compared with the control group. To conclude, exercise can promote extracellular matrix increased by production by increasing the levels of proteoglycan, aggrecan, and collagen type II in the degenerative intervertebral disc.
3.Correlation between the MRI-based grading system and F wave as well as H-reflex in patients with lumbar disc herniation
Xiao LI ; Caina LIN ; Haijie LUO ; Qing WAN ; Yuting RUAN ; Xinsheng ZHANG ; Shaoling WU ; Chao MA
Chinese Journal of Tissue Engineering Research 2016;20(42):6343-6350
BACKGROUND:Lumbar spine MRI and electrophysiological test are reliable methods for evaluating nerve root injury caused by lumbar disc herniation.
OBJECTIVE:To analyze the correlation between the MRI-based grading system and the latency and frequency of F wave as wel as latency and amplitude of H-reflex in patients with lumbar disc herniation.
METHODS:MRI imaging of the lumbar spine was performed with a 3.0-T imager and a dedicated TCL coil to classify lumbar disc herniation and nerve root compression. F wave and H reflex were detected on the patient bilateral tibial nerves using Oxford myoelectricity evoked potential instrument.
RESULTS AND CONCLUSION:Spearman correlation analysis showed that the MRI-based grading of patients with lumbar disc herniation had a negative correlation with F wave frequency (r=-0.594 0, P<0.000 1), and a positive correlation with F wave latency (r=0.825 6, P<0.000 1) and H-reflex latency (r=0.875 0, P<0.000 1), but no correlation with H-reflex amplitude (R=0.117 4, P=0.257 3). With MRI grading increased, F wave frequency was decreased, and F wave and H-reflex latency were prolonged gradual y, indicating aggravating nerve root compression.
4.Animal experiment and clinical study of Argon plasma coagulator in simple enucleation for small renal cell carcinoma
Changwen ZHANG ; Yong XU ; Kuo YANG ; Baojie MA ; Baomin QIAO ; Ting ZHANG ; Wenguang FAN ; Yan LIU ; Haijie XIE ; Yang SHI
Chinese Journal of Urology 2010;31(10):662-665
Objective To evaluate the feasibility and clinical effect of Argon plasma coagulator in simple enucleation for small renal cell carcinoma. Methods On the basis of successful performing the animal experience of coagulating therapy on the wound tissue during partial nephrectomy with Argon plasma coagulator in rabbit models, 10 cases of simple enucleation for small renal cell carcinoma with Argon plasma coagulator were accomplished. Results Both with the standard of stopping bleeding of wound tissue by Argon plasma coagulator and with the standard of re-spraying the wound tissue for 2 s after stopping bleeding using Argon plasma coagulator, the depth of wound tissue necrosis without blocking the renal pedicle is deeper than that with blocking the renal pedicle(P=0. 012 and P=0. 002, respectively).If the wound tissue was re-sprayed for 2 s after stopping bleeding by Argon plasma coagulator, the depth of the wound tissue necrosis without blocking the renal pedicle was deeper than that just with blocking the renal pedicle(P=0. 007 and P=0. 002,respectively). In the part of application in clinical, all procedures were successfully completed. The mean operative time was 163 min (range, 100-210 min) and mean blood loss was 230 ml (range, 100-400 ml). Drainage tube was pulled out 1 month after operation in 1 case for being allergic to absorbable hemostatic gauze, and the mean pulling drainage tube out time in others was 4. 2 d (range, 3-5 d). During a mean follow-up of 22 months (range, 10-38 months), no local tumor recurrence and distant metastasis was found. Conclusion Argon plasma coagulator can be used in simple enucleation for small renal cell carcinoma, and the clinical effectiveness is ideal.
5.Proanthocyanidins inhibit pancreatic cancer AsPC-1 cell growth and migration through up-regulation of let-7a.
Jia MA ; Binbin FANG ; Cong MA ; Haijie PANG ; Fanpeng ZENG ; Jun XIA
Journal of Southern Medical University 2015;35(8):1110-1115
OBJECTIVETo ascertain whether proanthocyanidins inhibit cell growth and migration by increasing let-7a expression in pancreatic cancer AsPC-1 cells.
METHODSThe proliferation rate, cell apoptosis rate and cell migration ability of AsPC-1 cells treated with proanthocyanidins were measured by MTT assay, Annexin V-FITC/PI staining, and Transwell migration assay, respectively. The expression of let-7a AsPC cells was detected by miRNA real-time RT-PCR after proanthocyanidins treatment. The changes in the biological behaviors of AsPC-1 cells were evaluated after transfection with let-7a mimics.
RESULTSCompared with the control group, proanthocyanidins treatment caused dose-dependent decrements of the proliferation rate and migration ability and increased the apoptosis rate in AsPC-1 cells. AsPC-1 cells with proanthocyanidins treatment showed increased expression of let-7a. Transfection with let-7a mimics resulted in obvious decreases in the cell growth rate and migration ability, and proanthocyanidins treatment significantly enhanced the inhibitory effect of let-7a mimics.
CONCLUSIONProanthocyanidins-induced cell growth and migration inhibition are partially mediated by up-regulation of let-7a expression in AsPC-1 cells.
Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Humans ; MicroRNAs ; metabolism ; Pancreatic Neoplasms ; pathology ; Proanthocyanidins ; chemistry ; Transfection ; Up-Regulation
6.Grape seed proanthocyanidins extract inhibits pancreatic cancer cell growth through down-regulation of miR-27a expression.
Jia MA ; Binbin FANG ; Fanpeng ZENG ; Haijie PANG ; Cong MA ; Jun XIA
Journal of Central South University(Medical Sciences) 2015;40(1):46-52
OBJECTIVE:
To explore the eff ect of grape seed proanthocyanidins extract (GSPE) on the growth of pancreatic cancer cells and the underlying mechanisms.
METHODS:
The pancreatic cancer AsPC-1 cells were cultured in vitro. The effects of GSPE on cell proliferation, apoptosis and migration were analyzed by MTT, Annexin V-FITC/PI and Transwell migration assay, respectively. The expression of miR-27a and FOXO1 in AsPC-1 cells was determined by real-time RT-PCR and Western blot, respectively. The miR-27a inhibitors were applied to verify the role of miR-27a in mediation of GSPE effects.
RESULTS:
GSPE inhibited cell growth in a dose-dependent manner. This inhibitory effect was significant when the dosage of GSPE was more than 50 μg/mL (P<0.05 vs control). GSPE also could induce apoptosis and inhibit cell migration. MiR-27a expression was notably down-regulated when the dosage of GSPE was 75 μg/mL (P<0.01 vs control). Compared with the control group, cell proliferation inhibition was significantly increased in the miR-27a inhibitor group, the GSPE group and the miR-27a inhibitor plus GSPE group (P<0.01), while cell migration was significantly decreased (P<0.01). Compared with the GSPE or the miR-27a inhibitor group, the growth and migration inhibitory effects in the miR-27a inhibitor plus GSPE group were more obviously (P<0.01). Both GSPE and miR-27a inhibitor alone could up-regulate FOXO1 expression. But these effects were more apparent when they are applied in combination.
CONCLUSION
GSPE inhibites AsPC-1 cells' growth and migration partly through down-regulation of miR-27a expression.
Apoptosis
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Cell Line, Tumor
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drug effects
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Cell Movement
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Cell Proliferation
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Down-Regulation
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Grape Seed Extract
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pharmacology
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Humans
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MicroRNAs
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genetics
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metabolism
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Pancreatic Neoplasms
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pathology
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Proanthocyanidins
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pharmacology
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Up-Regulation
7.The accuracy of tumor size evaluation on invasive breast cancer based on cone beam breast CT
Yue MA ; Zhaoxiang YE ; Aidi LIU ; Lu YIN ; Peng HAN ; Haijie LI
Chinese Journal of Radiology 2019;53(4):286-291
Objective To evaluate the accuracy of cone?beam breast CT (CBBCT) on tumor sizing in patients with invasive breast carcinoma and analyze the influence factors. Methods The preoperative CBBCT images of 82 female patients (85 breast lesions) with invasive breast carcinoma confirmed by pathology from November, 2011 to December, 2017 in Tianjin Medical University Cancer Hospital were included in this retrospective study. All the patients underwent the pathology and immunohistochemical test after operation. Tumor size estimation were performed on preoperative CBBCT images. Referring to tumor size measurement on pathology, all the lesions were divided into two groups. Concordance was defined as the discrepancy in diameter less than 0.5 cm, and the discordance was more than 0.5 cm. Pathology examination was performed after resection, and estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER?2) and Ki?67 result were recorded. All the lesions were classified into molecular subtype, including 14 Luminal A, 50 Luminal B, 11 HER?2?enriched and 10 triple?negative. Intraclass correlation coefficient (ICC) and Pearson correlation coefficient were used to analyze the reliability of CBBCT on tumor sizing. CBBCT?pathology discordance was analyzed based on the clinical, histopathology and CBBCT features by using t test, Chi?square and Fisher exact test. ROC curve was used to analyze the cut?off value between tumor size and CBBCT?pathology discordance. Results The agreement between CBBCT (2.155 ± 0.799) cm and pathology (1.986 ± 0.933) cm measurement was on moderate degree based on the ICC value (ICC=0.781, P<0.01) and had positive correlation (r=0.803, P<0.01). CBBCT?pathology concordance was found in 71 lesions, and discordance in 14 lesions. The factors of family history, symptom, pathology type, molecular subtypes, histological grade, surrounding fat invasion, lymphatic invasion, axillary lymph node metastasis, HER?2 positive and Ki?67 high expression had no significant difference between two groups. ER or PR positive had significant difference, suggesting that the accuracy of evaluation on ER or PR negative lesions could be reduced. The cut?off value of ROC curve between tumor size and CBBCT?pathology discordance was 2.08 cm, and the area under curve was 0.70. In 85 lesions. 66 of 75 mass lesions and 5 of 10 non?mass lesions were consistent. The lesion type had significant difference between two groups (χ2=6.705, P=0.010), which suggested the CBBCT evaluation on non?mass could have discrepancy with pathology. Conclusion CBBCT has high accuracy on tumor size evaluation on invasive carcinoma. ER or PR negative, large lesions or non?tumor lesions should be alert to the impact of CBBCT?pathology discordance before surgery which may cause the alteration of surgical protocols.
8.Effect of miR-23b on the malignant phenotype and the sensitivity of lenvatinib in human hepatocellular carcinoma cells
Wanneng YAN ; Shibo LI ; Haijie MA ; Dongdong CHEN ; Jie WANG ; Ting LE ; Guoqiang ZHANG
Chinese Journal of Hepatology 2021;29(5):433-438
Objective:To investigate the effect of miR-23b on the malignant phenotype and the sensitivity of lenvatinib in human hepatocellular carcinoma cells.Methods:Human hepatocellular carcinoma cell line HepG2, SMMC-7721 and QGY-7703 were transfected with miR-23b mimic and its control, respectively. CCK-8 and EdU assay were used to detect cell proliferation. Transwell assay were used to detect changes in cell migration and invasion. Tube formation assay were used to detect vasculogenic mimicry formation. The comparison of the mean between groups was analyzed by t-test.Results:CCK-8 results showed that the A values ??of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were 0.325 ± 0.011 and 0.537 ± 0.026, respectively, which were significantly lower than the control group 0.430±0.017 and 0.752 ± 0.051 ( P < 0.05). Transwell assay result showed that the number of cell migration of human hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group was (517.220 ± 32.873) and (242.327 ± 20.793), respectively, which were significantly lower than that of the control group (724.130 ± 15.142) and (424.432 ± 27.212) ( P < 0.01). Simultaneously, the number of cell invasions in the miR-23b mimic group were (55.671 ± 7.514) and (64.670 ± 6.011), respectively, which were significantly lower than those in the control group (124.320 ± 11.782) and (156.204 ± 12.501) ( P < 0.01). Tube formation assay showed that the number of tube forming branches of hepatocellular carcinoma cell line QGY-7703 and SMMC-7721 in the miR-23b mimic group was (489.824 ± 42.035) and (435.201 ± 44.143), respectively, which were significantly lower than that of the control group (878.620 ± 31.618) and (785.430 ± 38.723) ( P ??< 0.01). In addition, EdU results showed that after miR-23b combined with lenvatinib, the positive rates of EdU staining of hepatocellular carcinoma cell line HepG2 and SMMC-7721 in the miR-23b mimic group were (32.905 ± 1.342)% and (24.811 ± 0.820)%, respectively, which were significantly lower than the control group (52.623 ± 2.441)% and (38.702 ± 1.312)% ( P < 0.05). Conclusion:miR-23b can inhibit the proliferation, migration, invasion and vasculogenic mimicry formation, and enhance the sensitivity of lenvatinib drug in human hepatocellular carcinoma cells.
9.Personal glucose meters coupled with signal amplification technologies for quantitative detection of non-glucose targets:Recent progress and challenges in food safety hazards analysis
Feng HE ; Haijie WANG ; Pengfei DU ; Tengfei LI ; Weiting WANG ; Tianyu TAN ; Yaobo LIU ; Yanli MA ; Yuanshang WANG ; A.m.abd EL-ATY
Journal of Pharmaceutical Analysis 2023;13(3):223-238
Ensuring food safety is paramount worldwide.Developing effective detection methods to ensure food safety can be challenging owing to trace hazards,long detection time,and resource-poor sites,in addition to the matrix effects of food.Personal glucose meter(PGM),a classic point-of-care testing device,possesses unique application advantages,demonstrating promise in food safety.Currently,many studies have used PGM-based biosensors and signal amplification technologies to achieve sensitive and specific detection of food hazards.Signal amplification technologies have the potential to greatly improve the analytical performance and integration of PGMs with biosensors,which is crucial for solving the challenges associated with the use of PGMs for food safety analysis.This review introduces the basic detection principle of a PGM-based sensing strategy,which consists of three key factors:target recog-nition,signal transduction,and signal output.Representative studies of existing PGM-based sensing strategies combined with various signal amplification technologies(nanomaterial-loaded multienzyme labeling,nucleic acid reaction,DNAzyme catalysis,responsive nanomaterial encapsulation,and others)in the field of food safety detection are reviewed.Future perspectives and potential opportunities and challenges associated with PGMs in the field of food safety are discussed.Despite the need for complex sample preparation and the lack of standardization in the field,using PGMs in combination with signal amplification technology shows promise as a rapid and cost-effective method for food safety hazard analysis.
10.Progress in the Study of Spindle Assembly Checkpoint in Lung Cancer.
Xinchen QIN ; Yao ZHANG ; Haijie YU ; Lijuan MA
Chinese Journal of Lung Cancer 2023;26(4):310-318
Spindle assembly checkpoint (SAC) is a protective mechanism for cells to undergo accurate mitosis. SAC prevented chromosome segregation when kinetochores were not, or incorrectly attached to microtubules in the anaphase of mitosis, thus avoiding aneuploid chromosomes in daughter cells. Aneuploidy and altered expression of SAC component proteins are common in different cancers, including lung cancer. Therefore, SAC is a potential new target for lung cancer therapy. Five small molecule inhibitors of monopolar spindle 1 (MPS1), an upstream component protein of SAC, have entered clinical trials. This article introduces the biological functions of SAC, summarizes the abnormal expression of SAC component proteins in various cancers and the research progress of MPS1 inhibitors, and expects to provide a reference for the future development of lung cancer therapeutic strategies targeting SAC components.
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Humans
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Cell Cycle Proteins/metabolism*
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Spindle Apparatus/metabolism*
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Protein Serine-Threonine Kinases/metabolism*
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M Phase Cell Cycle Checkpoints/genetics*
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Lung Neoplasms/metabolism*