1.Bt transgenic crops for insect-resistance and modification of Bt protein and utilization of stacking strategy.
Chinese Journal of Biotechnology 2015;31(1):53-64
Insecticidal protein genes from Bacillus thuringiensis are currently the most widely used insect-resistant genes. They have been transferred to many crops for breeding and production. Among them, cotton, maize, potato and other insect-resistant crops are commercialized, creating considerable economic benefit. In this review, we summarized advances in identifying functional genes and transgenic crops for insect resistance, compared different strategies for enhancing vigor of insecticidal protein and utilizing gene stacking as well as listing valuable groups of stacked genes. In addition, the methods for multiple gene transformation was discussed.
Animals
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Bacterial Proteins
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genetics
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Crops, Agricultural
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genetics
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Endotoxins
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genetics
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Hemolysin Proteins
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genetics
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Insecta
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Plants, Genetically Modified
2.Influence of FOXC1 downregulating on biological behavior of nasopharyngeal carcinoma (NPC) 5-8F cell lines
Bolin CHEN ; Shijiang YI ; Peng LIU ; Lei OUYANG ; Xun LEI
The Journal of Practical Medicine 2016;32(4):527-530
Objective To investigate the Influence of FOXC1 downregulating on biological behavior of nasopharyngeal carcinoma (NPC) 5-8F cell lines. Methods Western blotting was used to detect the protein levels of FOXC1 in human immortalized nasopharyngeal cell lines NP69, nasopharyngeal carcinoma cell lines 6-10B with lower metastasis potential and nasopharyngeal carcinoma cell lines 5-8F with higher metastasis potential. siRNA was used to downregulate the expression of FOXC1 in 5-8F cell lines, and then ability of migration and invasion were observed. Results FOXC1 protein expression level in NP69 cell lines, 6-10B cell lines and 5-8F cell lines were (0.27 ± 0.04), (0.7 ± 20.06), (1.08 ± 0.05) respectively, and the difference was statistically significant (P < 0.05). Also ability of cell migration and invasion significantly weakened after FOXC1 depletion in 5-8F cell lines (P < 0.05). Conclusion FOXC1 might regulate invasion and metastasis of nasopharyngeal carcinoma through changing expression of Fibronectin and Vimentin, and FOXC1 may be an available target for molecular target therapy of nasopharyngeal carcinoma.
3.The function and mechanism of ErbB3 and IGF1R in breast cancer with Herceptin resistance
Ruixin ZHANG ; Min DENG ; Bolin LIU ; Kai LUO ; Zhimin HE
Journal of Chinese Physician 2017;19(2):173-176,181
Objective To investigate the role of epidermal growth factor receptor 3 (ErbB3) and insulin-like growth factor-1 receptor (IGF1R) in enhancing the resistance of Herceptin in human breast cancer.Methods HRG (Heregulin,the ligand of ErbB3) or IGF2 (insulin-like growth factor2,the ligand of IGF1R) was correspondingly added into breast cancer cells SKBR3 and BT474,and then 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and were performed in these cells to evaluate the sensitivity of these cells to Herceptin.Furthermore,we used HRG or IGF2 antibodies to inhibit their joint receptors in Herceptin-resistant breast cancer cells SKBR3/POOL2 and BT474/HR20.Finally,the sensitivity of these treated cells to Herceptin was detected via MTS assay.HRG or IGF2 was added into breast cancer cell BT474,and co-IP assay was used to detect the expressions of ErbB3 and IGF1R which combined with ErbB2.Results The treatment groups used HRG or IGF2 enhanced the resistance of Herceptin in Herceptin-sensitive breast cancer cells.On the other hand,we used antibodies of HRG and IGF2 to block their combining with their receptors in Herceptin-resistant breast cancer cells,the cells became more sensitive to Herceptin.BT474 cell was treated with HRG or IGF2.The expressions of ErbB3 and IGF1R which combined with ErbB2 were increased.Conclusions The formation of heterodimers ErbB2/ErbB3 and ErbB2/IGF1R might enhance the resistance of Herceptin in ErbB2-overexpression human breast cancers.
4.Research on Seasonal Variation of Plasma Metabolic Markers of Wei-qi Deficiency
Cuihuan YAN ; Yali WANG ; Xinguo WANG ; Mingquan ZHANG ; Xiang LIU ; Bolin LI ; Wenli CAO
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(1):66-70
Objective To study the plasma metabolomics features of SD rats with wei-qi deficiency respectively in spring, summer, autumn and winter; To seek the potential markers related to wei-qi deficiency; To discuss the essence of wei-qi deficiency.MethodsTwenty male SD rats were adaptively fed 7 days before Chinese Vernal Equinox Day, Summer Solstice, Autumn Equinox and Winter Solstice, synchronous controled with temporal condition. The rats were randomly divided into experimental and control groups according to random number method. Rats in the experimental groupwere made into wei-qi deficiency models through fatigue stimulation alternately with cold and heat stimulation. Blood was collected at 12:00 each solar term. High performance liquid chromatography- mass spectrometry instrument was used to detect plasma metabolites. Partial least squates-discriminant analysis was used to process statistical analysis of the data to compared the original data of plasma metabolomics between the two groups of each season, and explore the difference metabolic markers between each two groups, then speculate the potential biomakers of wei-qi differency.ResultsWith the changes of spring, summer, autumn and winter, significant differences were showed in metabolic profile between the experimental and control groups. Homocysteine showed a significant difference in spring, summer and autumn; Ceramide showed a significant difference in summer and winter; Testosterone showed a significant difference in spring and summer; Cyclic guanosine monophosphate showed a significant difference in autumn and winter; Sarcosine showed a significant difference in spring, summer and winter. The physiological functions of these substances related to lipid metabolism, amino acid metabolism, neurotransmitter and hormone regulation.ConclusionPotential biomakers of wei-qi deficiency changed along with the seasonal variation, which mainly reflected in lipid metabolism, amino acid metabolism, neurotransmitter and hormone regulation.
5.Radiation dose rate received by patients with coronary heart disease during interventional management:comparison between fluoroscopy and movies
Weibin LIU ; Lianjun HUANG ; Jiufang GUO ; Yihui WANG ; Guoliang JIN ; Yujia LI ; Tao SHEN ; Bolin ZHANG ; Shen ZHANG ; Jian CHEN
Journal of Interventional Radiology 2015;(6):524-527
Objective To determine the real time radiation dose rate displayed on DSA equipment in interventional management for patients with coronary heart disease in order to provide medical staff with useful basis to reasonably control the radiation dose. Methods A total of 30 patients with coronary heart disease, who received interventional treatment at authors’ hospital in September 2014 with a GE DSA unit, were randomly selected. Intraoperative radiation dose rates when fluoroscopy was converted to movies , which were displayed on DSA at respective sequence, were determined, and the results were analyzed and compared. Results The fluoroscopy radiation dose rate ranged from 18.5 to 212.0 mGy/min, the mean value was (114.7±42.1) mGy/min;while the radiation dose rate of movies varied from 216 to 1 691 mGy/min, with a mean value of(970.1±298.4) mGy/min. The overall mean fluoroscopy-to-movies radiation dose ratio was 1 ∶8.5. The larger the digital flat panel detector area was , the bigger the mean radiation dose rate would be and the more obvious increase in the mean radiation dose rate of movies than that of the fluoroscopy would be observed; the mean radiation dose rate of fluoroscopy was lower than that of movies , and the difference was significantly. Conclusion During the performance of coronary intervention , the medical staff should make reasonable choice of the equipment and closely observe the dynamic real-time display of the radiation dose rate, promptly adjust the inspection mode and the controllable parameters when it is needed , make immediate evaluation of dose level that might cause radiation injury to the patient , and reasonably control the radiation dose to reduce the effects of ionizing radiation on human health.
6.Effect of cuecumin on mammalian target of rapamyein signaling pathway during ischemia-reperfusion injury in isolated rat lungs
Bolin REN ; Meng WANG ; Xiaofei LIU ; Xihua LU
Chinese Journal of Anesthesiology 2018;38(12):1525-1529
Objective To evaluate the effect of curcumin on the mammalian target of rapamyein (mTOR) signaling pathway during ischemia-reperfusion (I/R) injury in isolated rat lungs.Methods Sixty-four clean-grade healthy male Sprague-Dawley rats,aged 3-4 months,weighing 250-320 g,were divided into 4 groups (n=16 each) using a random number table method:sham operation group (S group),I/R group,low-dose curcumin group (LC group) and high-dose curcumin group (HC group).The rats only received in vitro perfusion without ischemia in S group.Isolated rat lungs were subjected to 60 min of ischemia followed by 75-min reperfusion to establish the lung I/R injury model in I/R group.Curcumin 5 and 10 μmol/L were added to perfusion fluid from the beginning of reperfusion in LC and HC groups,respectively.Airway resistance (Res),lung compliance,perfusion flow (Flow) and pulmonary venous partial pressure of oxygen (PaO2) were recorded at 10 min of first perfusion (T0) and 15,45 and 75 min of reperfusion (T1-3).Wet/dry lung weight ratio (W/D ratio) was measured at the end of reperfusion.The morphological structure and ultrastructure of lung tissues were observed by using a light microscope and a transmission electron microscope,respectively.The expression of mTOR,Tau protein,nuclear factor kappa B (NF-κB) and tumor necrosis factor-alpha (TNF-α) mRNA in lung tissues was detected by real-time polymerase chain reaction.The expression of mTOR,phosphorylated Tau protein (pS396 Tau protein),NF-κB and TNF-α protein in lung tissues was determined by Western blot.Results Compared with S group,Res at T1-3 and W/D ratio at T3 were significantly increased,lung compliance,Flow and PaO2 were decreased at T1-3,and the expression of mTOR,NF-κB and TNF-α protein and mRNA,Tau protein mRNA and pS396 Tau protein was up-regulated at T3 in I/R,LC and HC groups (P<0.05).Compared with I/R group,Res at T1-3 and W/D ratio at T3 were significantly decreased,lung compliance,Flow and PaO2 were increased at T1-3,and the expression of mTOR,NF-κB and TNF-α protein and mRNA,Tau protein mRNA and pS396 Tau protein was down-regulated at T3 in LC and HC groups (P<0.05).Compared with LC group,Res at T1-3 and W/D ratio at T3 were significantly decreased,lung compliance,Flow and PaO2 were increased at T1-3,and the expression of mTOR,NF-κB and TNF-α protein and mRNA,Tau protein mRNA and pS396 Tau protein was down-regulated at T3 in HC group (P<0.05).The microscopic examination showed that the injury to lung tissues was significantly attenuated in LC and HC groups as compared with I/R group.Conclusion The mechanism by which curcumin reduces I/R injury in isolated rat lungs is related to inhibiting mTOR signaling pathway.
7.Monitoring of CT dose and analysis of radiological protection in CT modules applied for COVID-19
Ansheng LIU ; Bolin HUANG ; Cuiling LI ; Kai YANG ; Hanqi DU ; Guilin YI
Chinese Journal of Radiological Medicine and Protection 2020;40(5):338-342
Objective:To investigate the safety and protection level of radiological treatment in the CT modules in the makeshift hosptials in Wuhan during the prevention and treatment of COVID-19 cases.Methods:The layout of the CT modules in makeshift hospitals, radiological protection facilities and personal protective equipment were investigated. Based on the national standards, the CT dose index was estimated and the radiological protection level at the CT modules were measured.Results:The layout of the CT modules in makeshift hospitals is reasonable, with well-equipped radiological protection facilities. Of 23 CT modules, 20 were up to standards with acceptability of 87.0%. The other three were unqualified each with 1 detection points having values in excess of the national standards. Which, after being modified immediately reached the national standards. In addition, CT dose index for 7 CT modules were estimated, with CTDI W within ±7.5%. Conclusions:The CT modules in Wuhan meet the requirements of radiological safety and protection during the prevention and treatment of COVID-19 cases.
8.Isolation and identification of primary brain microvascular endothelial cells of rat using thin-layer cell culture method
Bolin LI ; Fan ZHANG ; Haiqin LIU ; Yuanyu TANG
Chinese Journal of Neuroanatomy 2023;39(6):709-713
Objective:To extract rat brain microvascular endothelial cells(BMECs)using thin-layer cell culture method.Methods:The brain cortex of four-week-old SD rats was obtained,which was chopped,sieved,and digested with type II collagenase to obtain microvascular segments.The amount of culture medium was strictly controlled,and it was inoculated into culture flasks for primary culture.The target cells were morphologically observed by using an invert-ed phase contrast microscope,and the factor Ⅷ related antigen(FⅧRag)was identified by using immunocytochemi-cal staining.Results:The target cells cultured in the inverted phase contrast microscope showed typical endothelial-like monolayer cobble stone-like mosaic growth;FⅧRag positive cells accounted for more than 99%of the total cells.Conclusion:Thin-layer cell culture method can successfully isolate and cultivate high-purity rat BMECs.
9. Clinical significance of cytogenetic monitoring in chronic myeloid leukemia
Chengyun PAN ; Na XU ; Bolin HE ; Rui CAO ; Libin LIAO ; Changxin YIN ; Yangqing LAN ; Ziyuan LU ; Jixian HUANG ; Jin SUN ; Ru FENG ; Qifa LIU ; Xiaoli LIU
Chinese Journal of Hematology 2017;38(2):112-117
Objective:
To analyze the association of cytogenetic abnormalities with the prognosis of chronic myeloid leukemia (CML) patients in tyrosine kinase inhibitors (TKI) era.
Methods:
Karyotype analysis of chromosome G-banding was carried out in 387 newly diagnosed CML patients by short-term culture of bone marrow cells. The correlation of cytogenetic abnormalities and CML progression was explored in combination with ABL tyrosine point mutations.
Result:
Of 387 patients with positive BCR-ABL fusion gene assayed by fluorescence in situ hybridization (FISH) technique, 94.1% (364/387) patients were Ph positive and 5.9% (23/387) Ph negative; 320 patients (87.9%) had a translocation t (9;22) (q34;q11) and 5 (1.4%) a variant translocation t (v;22) . Additional cytogenetic aberrations (ACA) at diagnosis were found in 10.7% (39/387) Ph+ patients, major route ACA in 22 (56.4%) cases and minor route ACA in 15 (38.5%) cases and 2 patients (5.1%) lacked the Y chromosome (−Y) ; 23.4% (71/303) patients occurred ACA during TKI treatment and the most frequent abnormalities were abnormal chromosome numbersd, which were likely associated with high proportion of disease progression (
10. Clinical analysis of adult Philadelphia chromosome-positive acute lymphoblastic leukemia with p16 gene deletion
Bolin HE ; Na XU ; Yuling LI ; Chengyun PAN ; Rui CAO ; Libin LIAO ; Changxin YIN ; Yangqing LAN ; Ziyuan LU ; Jixian HUANG ; Hongsheng ZHOU ; Qifa LIU ; Xiaoli LIU
Chinese Journal of Hematology 2017;38(3):204-209
Objective:
To investigate the clinical implications of p16 gene deletion in adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) .
Methods:
Retrospective analysis of clinical, immunophenotypic, cytogenetics, molecular characteristics and prognosis of 80 newly diagnosed Ph+ ALL patients with p16 deletion.
Results:
Of 80 adult Ph+ ALL, the prevalence of p16 gene deletion was 31.3%. p16 gene deletion carriers frequently accompanied with high WBC counts (WBC≥30×109/L) and CD20 expression. The incidence of complex chromosome abnormality in p16 gene deletion group was higher than that in non-deletion group, with alternations in chromosome 7, 8, 19 and der (22) more frequently observed. There was no difference occurred between patients with or without p16 gene deletion in complete remission (CR) rate following induction chemotherapy combined with tyrosine kinase inhibitors (TKIs) . However, after three cycles of chemotherapy, the MMR and CMR rate in the p16 gene deletion group was lower than patients with wild-type p16 gene (