1.Preventive effect of nano-powder of Wugong-sanqi on postoperative intestinal adhesion of rats
Xin HUANG ; Yiming LI ; Fuqin XU ; Mingqian HE ; Ziting QIU ; Feihong BING
International Journal of Traditional Chinese Medicine 2013;(6):513-515
Objective To investigate the preventive effect of nano-powder of Wugong-sanqi (NW),the rhizome of Anemone flaccid,on postoperative intestinal adhesion in rats.Methods Fifty SD rats were subjected to operation with Ellis' method for establishing intestinal adhesion models,then randomly divided into 5 groups (n=10),namely model,positive (Dexamethasone,i.m.10 mg/kg),NW high,medium and low dose group (p.o.450,225 and 112 mg/kg,respectively).Another ten normal rats were selected as normal control group.After administration 3 days pre-operation and 7 days post-operation,all of rats were killed,the intestinal adhesion was graded and the tissues were observed by optical microscope.Results NW evidently reduced the severity of postoperative adhesion (P<0.05 or P<0.01),compared with model group.The histopathologic changes such as proliferation of fibroblast cells and capillary,interstitial granulomas and inflammatory cells infiltration in intestinal tissues were also improved significantly in NW groups.Conclusion NW could inhibit the formation of postoperative intestinal adhesion effectively.
2.Role of JAK-STAT signaling in the interstitial fibrosis of unilateral ureter obstruction mice
Fang WANG ; Niansheng YANG ; Mingqian LUO ; Rong LI ; Lili ZHANG ; Shuang WANG ; Rui ZHANG ; Xueqing YU
Chinese Journal of Nephrology 2008;24(3):168-173
Objective To study the role of JAK-STAT singal transduction pathway in the interstitial fibrosis of unilateral ureter obstruction (UUO)mice. Methods Mice UUO model was established and the phosphorylation of JAK-STAT was examined at day 1,4,7 and 14 after ligation of the ureter.Mice in the treatment group were treated with daily injection of selective JAK2 inhibitor AG490 starting 2 h before ureter ligation until sacrifice while vehicle alone was given to mice in the model control group.Mice were sacrificed at day 14 after the establishment of model.Renal tubular lesion and interstitial fibrosis were assessed on paraffin section.Immunohistochemistry was used to detect renal macrophage infihration and α-SMA expression.The expression of collagen Ⅲ and MCP-1 mRNA was measured by RT-PCR.Phosphorylation of JAK2and STAT1 was examined by Western blotting. Results JAK2-STAT1 signaling transduction pathway was activated in UUO model.The activation of JAK2-STAT1 was closely correlated with the progression of renal injury,tubular histological lesions and interstitial fibrosis.AG490 treatment significantly inhibited the phosphorylation of JAK2 and STAT1 (P<0.01).AG490 treatment also significantly reduced tubular lesions[(21.7 ±1.7)% vs (49.4±1.0)%]and interstitial fibrosis(1.0±0.1 vs 2.3±0.2),α-SMA expression(0.9±0.1 vs 2.1±0.2)and maerophage accumulation[(13.3±1.6)cells/HPF vs (34.4±1.0)cells/HPF](all P<0.01).In addition,AG490 significantly inhibited the expression of collagen Ⅲ and MCP-1 mRNA. Conclusion JAK-STAT signaling plays an important role in renal tubulointerstitial inflammation and fibrosis.
3.Expression of Synaptohydin in Hippocampus of Exercise-fatigue Rats by Sinisan
Wei LI ; Lie KONG ; Shuang YU ; Shuo ZHENG ; Mingqian XU ; Rong ZHANG ; Yan LIU ; Xiaolan LIU ; Feng LI
Chinese Journal of Rehabilitation Theory and Practice 2009;15(8):739-741
Objective To observe the effect of Sinisan on the learning and memorial ability and the expression of synaptohydin in the hippocampus of exercise-fatigue rats. Methods Rats were made into the fatigue model by Exhaustive Swimming. Learning and memorial ability of the rats were observed by moving-back with current stimulation experiment; expression of synaptohydin protein in hippocampus of exercise-fatigue rats treated with Sinisan for 10 days were detected by immunohistochemistry technique and compared to control group and model group.Results The moving-back time and the error times of the exercise-fatigue rats as follows: model group(73.00±61.96) s/(1.67±1.15), control group(144.25±57.14) s/(0.50±0.80),Sinisan group (166.17±47.92) s/(0.38±0.49). Comparison of mean optical density of synaptohydin protein in hippocampus of rats in different groups as follows: control group (0.2636±0.10654) and Sinisan group (0.2555±0.163380) were higher than the model group (0.0474±0.1837)(P<0.05). The synaptohydin staining Results showed that synaptohydin-positive granules around the neuron in hippocampus of rats in control group were the darkest and densest, the Sinisan group were darker than the model group rats.Conclusion Sinisan could improve the learning and memorial ability and increase the synaptohydin protein in the hippocampus of the exercise-fatigue rats.
4.An anti-human ovarian carcinoma and CD3 bispecific single-chain antibody mediates CDR3 spectratype drift of T cell receptor alpha and beta chains.
Wei LUO ; Qian WEN ; Mingqian ZHOU ; Li MA
Journal of Southern Medical University 2012;32(7):919-923
OBJECTIVETo analyze the drift of T cell receptor (TCR) Vα and Vβ gene family CDR3 spectratype in response to ovarian carcinoma cells mediated by an anti-human ovarian carcinoma/CD3 bispecific single-chain antibody (BHL-1), and explore the mechanism of the bispecific single-chain antibody-mediated T cell immune response.
METHODSImmunoscopic spectratyping technique was used to analyze the TCR repertoire diversity (CDR3 spectratype distribution) of the T cells from 6 healthy donors before and after stimulation of the cells with human ovarian carcinoma in the presence of BHL-1. The predominant usage of TCR α and Vβ chain CDR3 was analyzed after the stimulation, and sequence analysis was performed for the CDR3 region of the monoclonal T cells.
RESULTSThe spectratypes of Vα and Vβ gene family TCR CDR3 region showed a Gaussian distribution before stimulation of the T cells from the 6 donors. After stimulation of the T cells, CDR3 spectratype drift occurred in the T cells, and some TCR Vα and Vβ families showed an anomalous and oligoclonal expansion. Different CDR3 sequences of the Vα and Vβ gene family TCR were found in the monoclonal T cells stimulated with BHL-1.
CONCLUSIONCDR3 spectratype drift occurs in TCR α and Vβ chains of T cells after stimulation with human ovarian carcinoma cells and BHL-1, indicating that the predominant usage of TCR Vα and Vβ families is associated with the specific T cell immune response mediated by BHL-1.
Antibodies, Bispecific ; immunology ; Cell Line ; Cell Line, Tumor ; Complementarity Determining Regions ; immunology ; Female ; Humans ; Monocytes ; immunology ; Ovarian Neoplasms ; immunology ; Receptors, Antigen, T-Cell, alpha-beta ; immunology ; Single-Chain Antibodies ; immunology
5.Effect of gene silencing of Bmi-1 on proliferation regulation of CD44+ nasopharyngeal carcinoma cancer stem-like cells.
Xinhua XU ; Yang LIU ; Daojun LI ; Jin SU ; Juan HU ; Mingqian LU ; Fang YI ; Jinghua RENG ; Weihong CHEN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(10):941-947
OBJECTIVE:
To investigate the effect of gene silencing of Bmi-1 on proliferation regulation of CD44+ nasopharyngeal carcinoma cancer stem-like cells (CSC-LCs).
METHOD:
The sequence-specific short hairpin RNA lentivirus targeting at human Bmi-1 gene (LV-Bmi-1shRNA) was constructed and was used to infect CD44+ nasopharyngeal carcinoma cells which were sorted by flow cytometry. A lentiviral which included a random sequence was also designed to serve as a negative control. We employed fluorescence microscope and flow cytometry to detect infection efficiency; real-time PCR was used to detect Bmi-1 and its downstream gene while each protein expression level was confirmed by western blotting protocol; CCK-8 proliferation assay was applied to measure proliferation capacity; tumor spheroid assay was used to evaluate the self-renewal capacity. Colony formation assay was used to measure cell colony formation capability; flow cytometry analyzed cell cycle distribution.
RESULT:
The constructed LV-Bmi-1shRNA successfully infected into the CD44+ nasopharyngeal carcinoma cells. The infection efficiency could reach above 95%; LV-Bmi-lshRNA effectively inhibited Bmi-1 mRNA and protein expression, while the downstream gene p16INK4a and p14ARF mRNA as well as protein expression level were upregulated (P < 0.05). Notablely, the proliferation, colony formation, self-renewal capabilities of the experimental group decreased significantly (P < 0.05). In addition, the cell cycle arrested at the G0-G1 phase.
CONCLUSION
Gene silencing of Bmi-1 inhibited the proliferation, colony formation and self-renewal capabilities of the CD44+ nasopharyngeal carcinoma CSC-LCs, inhibited the cell cycle processes, which may mediate through Bmi1-p16INK4a/p14ARF-p53 pathway. Our experimental results indicated that Bmi-1 gene may play an important role in the maintenance of the stem cell-like characteristics of CD44+ nasopharyngeal carcinoma cells. Bmi-1 gene may be a potential new target for the treatment of nasopharyng al carcinoma in the future.
Carcinoma
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Cell Cycle
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Cell Division
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Cell Line, Tumor
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Cyclin-Dependent Kinase Inhibitor p16
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metabolism
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Gene Silencing
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Humans
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Hyaluronan Receptors
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metabolism
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Lentivirus
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Nasopharyngeal Carcinoma
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Nasopharyngeal Neoplasms
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genetics
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pathology
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Neoplastic Stem Cells
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cytology
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Polycomb Repressive Complex 1
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genetics
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RNA, Messenger
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RNA, Small Interfering
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Tumor Suppressor Protein p14ARF
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metabolism
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Tumor Suppressor Protein p53
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metabolism
6.Application of preoperative and intraoperative electroneurophysiological examination for persons with hemifacial spasm
Ying HE ; Yuzuo LI ; Jing CHEN ; Mingqian PAN ; Yong ZHANG ; Xiaoyi LI
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(6):506-510
Objective:To analyze the diagnostic value of preoperative electromyography and spasticity assessment for patients with hemifacial spasm, and to define a relationship between intraoperative electrophysiological examination and prognosis in order to provide help for clinical diagnosis and treatment.Methods:Thirty-one patients with hemifacial spasm were selected for the clinical spasticity scoring and divided into a general spasm group ( n=27) and a severe spasm group ( n=4). All received preoperative neurophysiological examination to record their twitch discharge, facial nerve conduction velocity (MCV), lateral spread (LSR) of the spasm, brainstem auditory evoked potential (BAEP), and blink reflex. Electrophysiological monitoring then recorded intraoperative LSR. According to whether the LSR disappeared or not, the patients were divided into the LSR disappearance group (of 15) and the LSR residual group (of 16), and facial muscle activity was recorded again one, three and six months after the operation. Results:Preoperative EMG examination of both groups showed positive LSR and that facial nerve MCV was within the normal range. There were, though, significant differences between the two groups in the twitching discharge by needle electromyography, blink reflex and preoperative BAEP. One week after the operation, one member of the residual group and 3 from the disappearance patients of the former and latter group had recovered in terms of LSR, with 3 and 7 cases significantly relieved, respectively. Two months later, the corresponding figures were 5 and 7, 3 and 6, respectively. Half of a year after the surgery, 5 from the residual group and 12 from the disappearance group had fully recovered in terms of LSR, while 9 and 2 cases were significantly relieved. Altogether, there were significant differences within the two groups in terms of recovery among all the time points, with significantly better recovery in the LSR disappearance group than the LSR residual group at 1 week after operation, while there were no significant differences between the two groups in recovery 3 and 6 months after their operation.Conclusions:Preoperative electromyography can provide objective assessments of the scope, severity, and facial nerve excitability of patients with hemifacial spasm. Real-time intraoperative electrophysiology monitoring can help surgeons to objectively assess the effect of decompression and to find and avoid nerve traction injury in surrounding areas quickly.
7.Research Ideas and Evaluation of the Functional Material Base of Traditional Chinese Medicine Formula
Jianxun LIU ; Mingqian SUN ; Junguo REN ; Li LIN
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(4):473-478
The function of Traditional Chinese Medicine (TCM) represents the overall effect of the formula. The effect is the expression of numerous biological effects of chemical components of TCM in certain pathological conditions of the body function adjustment, and these chemicals are the material basis of the TCM function. Limited by many conditions, few studies have been done on the material base of TCM function, and most of them have been supplemented by the substance basis of TCM efficacy. This leads to the disconnection between the theory of traditional Chinese medicine and modern research, and it is difficult to scientifically explain the functional indications of Chinese medicine. This paper discusses and summarizes the ideas and methods of modern research and evaluation of the material base of TCM function.Under the guidance of TCM theory and clinical practice, we set up an index system for evaluating the TCM function, and further explained the material basis and action mechanism of TCM formula. Through the organic combination of multidisciplinary and multi-technology, it provides a reference for the establishment of a comprehensive research model of the functional material basis of TCM formula.
8.The research status and prospects of microRNA-glial regulatory network in radiation-induced brain injury
Mingqian OU ; Furong SUN ; Weihao FAN ; Lili CUI ; Minhua LI ; Meijun LIN ; Yangsheng YU ; Shiyun LIANG ; Haihong ZHOU
Chinese Journal of Radiological Medicine and Protection 2020;40(7):564-569
Radiation-induced brain injury (RBI) is the most serious complication of head and neck tumor after radiotherapy. The pathogenesis of RBI is complicated, and the clinical course is irreversible, while no effective treatment available. The activation of glial cells is one of the main theories of RBI, and the prevention and treatment of RBI by targeting glial cells is the focus of current research. As a post-transcriptional regulatory factor, microRNA (miRNA) has been confirmed to be involved in regulatingglial cell radiosensitivity, inflammation type transformation, autophagy, exosomatic, long non-coding RNA (lncRNA), circular RNA (circRNA) and other related pathways, thereby mediating the occurrence and development of cascade reaction of inflammatory injury and neurological function repair of central nervous system (CNS) disease. Therefore, the establishment of miRNA - glial regulatory network may provide a new strategy for the prevention and treatment of RBI.
9.Alkaloid profiling of the traditional Chinese medicine Rhizoma corydalis using high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry.
Mingqian SUN ; Jianxun LIU ; Chengren LIN ; Lan MIAO ; Li LIN
Acta Pharmaceutica Sinica B 2014;4(3):208-216
Since alkaloids are the major active constituents of Rhizoma corydalis (RC), a convenient and accurate analytical method is needed for their identification and characterization. Here we report a method to profile the alkaloids in RC based on liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS/MS). A total of 16 alkaloids belonging to four different classes were identified by comparison with authentic standards. The fragmentation pathway of each class of alkaloid was clarified and their differences were elucidated. Furthermore, based on an analysis of fragmentation pathways and alkaloid profiling, a rapid and accurate method for the identification of unknown alkaloids in RC is proposed. The method could also be useful for the quality control of RC.
10.Genomic, transcriptomic, and epigenomic analysis of a medicinal snake, Bungarus multicinctus, to provides insights into the origin of Elapidae neurotoxins.
Jiang XU ; Shuai GUO ; Xianmei YIN ; Mingqian LI ; He SU ; Xuejiao LIAO ; Qiushi LI ; Liang LE ; Shiyu CHEN ; Baosheng LIAO ; Haoyu HU ; Juan LEI ; Yingjie ZHU ; Xiaohui QIU ; Lu LUO ; Jun CHEN ; Ruiyang CHENG ; Zhenzhan CHANG ; Han ZHANG ; Nicholas Chieh WU ; Yiming GUO ; Dianyun HOU ; Jin PEI ; Jihai GAO ; Yan HUA ; Zhihai HUANG ; Shilin CHEN
Acta Pharmaceutica Sinica B 2023;13(5):2234-2249
The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.