1.Dual role of high mobility group box 1 (HMGB1) in tumor progression
Yueqi CHEN ; Zhaoliang SU ; Lin XIA
Chinese Journal of Microbiology and Immunology 2021;41(3):221-225
High mobility group box 1 (HMGB1), a ubiquitous non-histone protein in the nuclear chromatin of eukaryotic cells, plays an important role in inflammatory diseases, autoimmune diseases and cancer. The function of HMGB1 in the development and progression of tumor varies with its subcellular localization. On one hand, HMGB1 promotes the growth, proliferation, invasion and metastasis of tumor cells, and mediates immune escape and immune tolerance; on the other hand, HMGB1 possesses anti-tumor activity by maintaining the stability of genome, regulating the expression of tumor suppressor genes, inhibiting cell invasion and metastasis and improving the anti-tumor therapeutic effect. In recent years, increasing evidences have suggested that HMGB1 is closely associated with the development of tumor. Therefore, understanding the function of HMGB1 in cancer will provide enlightenment for preventive and therapeutic strategies. This review summarized the dual role of HMGB1 in tumor progression.
2.Effect of mycophenolate mofetilglial on glial scar formation into the hippocampus and learning and memory functions of the rats with diffuse axonal injury
Yiguo YIN ; Zhaoliang SUN ; Ertao CHEN ; Dongfu FENG
Chinese Journal of Trauma 2015;31(10):949-953
Objective To determine the intervention of mycophenolate mofetil (MMF) in glial scar formation and learning and memory function in a rat model of diffuse axonal injury (DAI).Methods Ninety-six SD rats were randomly divided into sham group, normal saline (NS) group and mycophenolate mofetil (MMF) group according to the random number table, with 32 rats per group.Immunohistochemistry was used to detect activated microglia cells, activated astrocytes and chondroitin sulphate proteoglycanns (CSPGs) in the hippocampus.Inage-Pro Plus software was used to quantitatively assess the changes of activated microglia cells, activated astrocytes and CSPGs.Morris water maze was applied for testing rat learning and memory function.Integrated absorbance (IA) of major constituents (microglia, astrosyte, chondroitin sulphate proteoglycan) of the glial scar was determined and analyzed for the correlation with the parameters of MWM.Results At 7, 14 and 28 days after injury, MMF group showed decreased IA of activated microglia in the hippocampus compared to sham and NS groups (P < 0.05).At 7-11 days after injury, percent distance and percent time in the target quadrant of Morris water maze did not differ significantly among the three groups and were not related to the IA of glial scar.At 28-32 days after injury, percent distance and percent time in the target quadrant of Morris water maze lowered significantly in MMF group.At 28 days after injury, IA of the glial scar had a positive correlation with mean speed and mean escape latency, but negative correlation with percent distance and time in the target quadrant that measured in Morris water maze at 28-32 days after injury.Conclusion MMF significantly attenuates glial scar formation into the hippocampus and improves learning and memory function in rats during the recovery stage when administered in the early stage after DAI.
3.Determination of Organic Acids from Mainstream Cigarette Smoke using Ultra Low Temperature Solvent Extraction Followed by Comprehensive Two-Dimensional Gas Chromatography/Time-of-Flight Mass Spectrometry
Quan ZHANG ; Zhangmin XIANG ; Kai CAI ; Shuping ZHOU ; Zhaoliang GENG ; Yushuang GUO ; Wensheng CHEN
Chinese Journal of Analytical Chemistry 2014;(8):1125-1131
A new method for the determination of organic acids from mainstream cigarette smoke using ultra low temperature solvent extraction-comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry(GC×GC-TOF/MS)was established. The mainstream smoke was directly trapped by ethyl ether with ultra low temperature solvent extraction device, and cleaned up with liquid-liquid extraction. The concentrated extracts were further derived by N,O-bis(trimethylsily)trifluoroacetamide (BSTFA). The good separation of silanized product was achieved by the column set of DB-1 (30 m × 0. 25 mm, 1. 0 μm) as the 1st column and DB-wax (1. 5 m × 0. 1 mm, 0. 1 μm) as the 2nd column with modulation period of 6 s and scan range of m/z 45-450 . The results showed that linearity correlation coefficients were larger than 0 . 99 , and the average recoveries were between 80. 17% and 107. 81% with the relative standard deviations (RSD) in the range of 0 . 4%-12 . 1% ( n=5 ) . The detection limit and the quantitation limit were 1 . 3-24 . 5 μg/kg and 4. 1-77. 1 μg/kg, respectively.
4.HLA-A2-restricted point-mutated Survivin peptide elicited specific cytotoxic T lymphocyte reaction against hepatocarcinoma
Han SHEN ; Zhaoliang HUANG ; Xiaohua CHEN ; Yanmei LIN ; Teng WANG ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Microbiology and Immunology 2012;32(8):696-701
Objective To investigate the anti-hepatocarcinoma(anti-HCC) function of HLA-A2-restricted point-mutated Survivin peptide induced CTLs.Methods The HLA-A2-restricted Survivin nonapeptides were evaluated using bioinformatics software.The binding affinity of Survivin peptide to HLA-A2 molecular was determined with flow cytometry analysis.After peptide-induced CTLs were generated in vitro,flow cytometry and ELISA were performed to detect the levels of IFN-γ,which were secreted by reactive CTLs.Peptide-induced CTLs were co-cultured with hepatoma cell lines HepG2 and BEL-7402.The rates of tumor cells lysis were assayed using CytoTox 96(R) and the morphological changes of tumor cells were observed with inverted phase contrast microscope.Results Point-mutated Survivin nonapeptide Sur79M2 (KMSSGCAFL) was filtered out,which was shown higher scores compared with the wild-type peptide Sur79.Consistent with the results of software analysis,Sur79M2 showed higher binding ability in T2 binding assays.At the same time,Sur79M2-induced CTLs could release a large number of IFN-γ after incubated with target cells rather than Sur79.When co-cultured with HCC cell lines HepG2 and BEL-7402,Sur79M2-induced CTLs effectively lysis HepG2 on HLA-A2-restricted manner without killing effect on BEL-7402 that do not express HLA-A2 molecules.Conclusion Sur79M2 could elicited specific cytotoxic T lymphocytes in vitro,which were able to specifically kill HCC cell lines on HLA-A2-restricted manner.
5.miR-592 induces apoptotic cell death in human U251 cells
Zhaoliang CHEN ; Hong LIU ; Huili YANG ; Yukai GAO ; Juan ZHANG ; Wenyu JI
Chinese Journal of Nervous and Mental Diseases 2017;43(4):234-238
Objective To investigate the role of miR-592 in the Glioma.Methods We first analyzed the expression of miR-592 in Glioma tissues from patients by quantitative PCR.We transfected U251 cells with miR-592 mimics and then detected the growth of cells by MTT assay.We performed dual-luciferase reporter assay and western blot assay to examine whether Runx2 was the direct target of miR-592 in U251 cells.In order to test whether Runx2 was the functional target of miR-592,we determined the cell growth curve by down-regulating the level of Runx2.Moreover,we also detected the apoptosis of U251 after Runx2 knockdown.Results The expression of miR-592 was significantly reduced in glioma tissues (t=2.752,P--0.013).Over-expression miR-592 remarkably increased the apoptotic rate of U251 cells compared with the control group (t=2.127,P=0.031;t=2.284,P=0.026).Flow cytometry analysis showed that MiR-592 significantly promoted apoptotic cell death of U251 cells Apoptosis rate was 7.2%±0.68% in miR-592 group and 17.47%±1.45% in control group (t=3.294,P=0.007).The results of double luciferase assay and Western blot assay showed that miR-592 directly targeted the 3'Runx2 of-UTR to inhibit the level of Runx2 protein.The effect of down-regulation of Runx2 on the growth of U251 cells was detected,the results showed that growth was significantly slower in the cells transfected with Runx2 siRNA than in those without Runx2 siRNA (t=3.124,P=0.O11).Detection of cycle by flow cytometry showed that runx2 down-regulated the apoptosis rate of U251 cells.Tumor growth curve showed that overexpression of miR-592 significantly inhibited tumor growth and the down regulation of Runx2 expresssion also significantly inhibited tumor growth.Conclusion miR-592 suppresses the growth and promotes the apoptotic rate of U251 cells by targeting Runx2.
6.Role of interleukin-17 in spinal dorsal horns in neuropathic pain in rats and its effect on activation of astrocytes
Caixia SUN ; Wen YUAN ; Chunye LI ; Yanfang LIU ; Pan CHEN ; Zhaoliang SU ; Huaxi XU
Chinese Journal of Anesthesiology 2015;35(3):320-325
Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.
7.IL-10 producing regulatory B cells ameliorated cardiac hypertrophy of viral myocarditis mice
Shiqing ZHANG ; Jia CHEN ; Lin XIA ; Zhaoliang SU ; Huaxi XU ; Fang LIU
Chinese Journal of Microbiology and Immunology 2021;41(5):345-352
Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.
8.Initial experience of transurethral enucleation of submucosal bladder leiomyoma
Jianjun SHA ; Jiahua PAN ; Zhaoliang WANG ; Lianhua ZHANG ; Wei CHEN ; Jianwei LV ; Jing LENG ; Juanjie BO ; Dongming LIU ; Yiran HUANG
Chinese Journal of Urology 2011;32(9):636-638
ObjectiveTo evaluate the clinical effecacy and safety of transurethral enucleation of submucosal bladder leiomyoma.MethodsAnalyze retrospectively the clinical data of 6 patients (2 male,4 female) of submucosal bladder leiomyoma. The mean age was 59 years (range 32- 78). The clinical manifestations included dysuresia in 3 cases, irritative bladder in 1 case, gross hematuria in 1 caes, and no clinical symptoms in 1 case. The mean course was 23 months (range 1 week-4 years). All the bladder tumors were indicated by ultrasonography, CT scan and cystoscopy, while 4 cases of bladder filling defects were showed by intravenous urogram. Before the tranaurethral enucleation of the bladder tumor, a deep needle biopsy and pathological examination were carried out to confirm the diagnosis of bladder leiomyoma. For the relatively small lateral leiomyomas, holium laser enucleation was carried out, while for the big ones, resectoscope enucleation was used to remove the mass. Biopsys were performed after complete removal of the tumor.ResultsTransurethral enucleation of 6 cases of submucosal bladder tumors were successful without any postoperative complication. All patients regained normal urination, and lower urinary tract irritation was relieved obviously and hematuria disappeared. No tumor recurrence or metastasis was found during the follow-up of 4 to 158 months.ConclusionsCystoscopy and transurethral biopsy are the most important methods for the diagnosis of submucosal bladder leiomyoma. The transurethral enucleation is a feasible and safe surgical technique for such patients with excellent results.
9.The distribution and structure of class Ⅰ integron in the multidrug-reisistant Pseudomonas aeruginosa
Jianguo CHEN ; Xiaoli DAI ; Yufeng JIANG ; Yingzhao LIU ; Jianren YU ; Zhaoliang SU ; Xinxiang HUANG ; Chiyu ZHANG ; Shengjun WANG ; Qixiang SHAO ; Shihe SHAO ; Huaxi XU
Chinese Journal of Laboratory Medicine 2008;31(8):872-875
Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.
10.Avian Influenza: Should China Be Alarmed?.
Zhaoliang SU ; Huaxi XU ; Jianguo CHEN
Yonsei Medical Journal 2007;48(4):586-594
Avian influenza has emerged as one of the primary public health concern of the 21st century. Influenza strain H5N1 is capable of incidentally infecting humans and other mammals. Since their reemergence in 2003, highly pathogenic avian influenza A (H5N1) viruses have been transmitted from poultry to humans (by direct or indirect contact with infected birds) in several provinces of Mainland China, which has resulted in 22 cases of human infection and has created repercussions for the Chinese economy. People have been concerned whether a new pandemic will occur in the future. The eradication of pathogenic avian influenza viruses appears to be the most effective way to prevent an influenza pandemic. This paper will examine the features of H5N1, including incidence, infection, immunity, clinical management, prevention and control, and therapy in Mainland China.
Adolescent
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Adult
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Animals
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Birds
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Child
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China/epidemiology
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Disease Outbreaks/prevention & control
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Female
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Humans
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Incidence
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*Influenza A Virus, H5N1 Subtype
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Influenza in Birds/prevention & control
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Influenza, Human/*epidemiology/immunology/therapy
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Male
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Zoonoses/epidemiology/transmission/virology