1.Study on histone H3 acetylation of BDNF gene promoter in SAMP8 mice
Chengzhan WANG ; Gengdi HUANG ; Qing FANG ; Jijian SI ; Jianli YANG
Chinese Journal of Behavioral Medicine and Brain Science 2016;25(4):295-299
Objective To explore the role of histone H3 acetylation modification of brain derived neurotrophic factor (BDNF) in the pathogenesis of Alzheimer's disease (AD).Methods 2 months and 8 months SAMP8 mice were used as AD model.Morris water maze was used to detect the impairment of learning and memory.Western blot was used to detect BDNF protein expression in the hippocampus,and chromatin immunoprecipitation (CHIP) was applied to study the changes of histone H3 acetylation in different BDNF promoters.Results The results of water maze test showed that the time across the target quadrant in 8 months SAMP8 mice(0.9±0.4) was significant declined compared with that of 2 months SAMP8 mice(3.7 ± ±0.9) and 8 months SAMR1 mice (3.3±0.6)(all P<0.05).Meanwhile,compared with 2 months SAMP8 mice ((23.9±4.0) s) and 8 months SAMR1 mice ((21.5± 2.3) s),target quadrant time in the 8 months SAMP8 mice((11.7±2.8) s) was also significantly reduced(both P<0.05).The western blot showed the expression of BDNF in the hippocampus of 8 months SAMP8 mice was significantly decreased compared with that of 2 months SAMP8 mice and 8 months SAMR1 mice(P<0.05).Lastly,CHIP assays showed that histone H3 acetylation of BDNF exon Ⅳ and Ⅵ in the hippocampus of 8 months SAMP8 mice were remarkably decreased(P<0.05) compared with that of 2 months SAMP8 mice and 8 months SAMR1 mice.There was no significant change of histone H3 acetylation of BDNF exon Ⅰ and Ⅲ among all groups(P>0.05).Conclusion Histone H3 acetylation of BDNF exon Ⅳ and Ⅵ is reduced during the development of AD,which may be the mechanism underlying the impairment of learning and memory in AD.
2.Clinical study of correlation of blood perfusion characteristics in liver cancer by contrast-enhanced ultrasonography and individualized interventional therapy
Qin SI ; Xiaoli QIAN ; Xushun LIU ; Yanli HUANG ; Zengcai LI ; Lu YANG
Chinese Journal of Ultrasonography 2010;19(11):948-951
Objective To identify the blood perfusion characteristics of different kinds of liver cancer and individualize interventional treatment models by color Doppler ultrasound(CDUS) and contrast-enhanced ultrasonography (CEUS). Methods The blood perfusion characteristics of 315 patients with liver cancer comfirmed by pathology were analyzed and classified using CDUS and CEUS. Individual interventional therapy models were selected based on the classification. Therapeutic effects were evaluated by EUS,contrast-enhanced CT(CECT) and DSA simultaneously. Results Liver cancer can be classified by the blood perfusion characteristics from CEUS examination. The individualized interventional treatment models were selected and evaluated according to the characteristics:①Solitary small HCC of diameter ≤3 cm group were treated via percutaneous ethanol injection(PEI),radiofrequency(RF) or percutaneous microwave coagulation therapy(PMCT). Necrosis rates of the lesions were 95.0% - 97.9%, 1 and 3 years survival rates were 98.0% and 87.8% ,respectively. ②Diameter ≤5 cm and lesions ≤3 group were treated with RF or PMCT combined PEI. Necrosis rates of the lesions were 93.7% - 94.8% ,1 and 3 years survival rates were 89.8% and 81.4% ,respectively. ③Diameter >5 cm and hypervascular tumor main fed by the hepatic artery group were treated with TACE,PEI combined RF or PMCT. Necrosis rates of the lesions were 71.4% - 73.8%, 1 and 3 years survival rates were 66.2% and 47.6%, respectively. ④ Diameter >5 cm and hypervascular tumor fed by double blood supply or accompanied by portal vein tumor thrombus group were treated with selective portal vein embolization(SPVE) based on the above treatments. Necrosis rates of the lesions were 53.3% - 55.6%, 1 and 3 years survival rates were 64.7% and 40.0%, respectively. Conclusions Classifying the blood perfusion characteristics and choosing individualized interventional treatment models by CEUS are of important clinical significance in non-surgical treatment of liver cancer.
3.The effects of neonatal MK-801 treatment on expression of NMDA receptors and NGF in rats
Jiao JIA ; Jitao LI ; Runhu HUANG ; Hong YANG ; Yunai SU ; Tianmei SI
Chinese Journal of Nervous and Mental Diseases 2013;(12):733-738
Objective To investigate the effects of repeated neonatal administration of dizocipline maleate (MK-801), the N-methyl-D-aspartate (NMDA) receptor antagonist, on the expression of NMDA receptor subunits NMDAR 1 (NR1), NMDAR2A (NR2A), NMDAR2B (NR2B) and the protein levels of nerve growth factor (NGF) in neonatal rats. Methods Neonatal Sprague-Dawley (SD) rats were randomly divided into research group and control group, with 15 ani-mals in each group. Rats were administrated subcutaneously with MK-801 or normal saline from postnatal day (PND) 5 to PND14 (0.25 mg/kg, twice a day). The expression levels of NR1, NR2A, NR2B and NGF were examined on PND15, PND42 and PND70 in the prefrontal cortex and hippocampus. Results At PND15 (neonatal period), there were no signifi-cant differences in the expression levels of NR1, NR2A, NR2B and NGF in the prefrontal cortex and hippocampus be- tween the two groups (P>0.05). At PND42 (adolescence), NGF protein levels in the prefrontal cortex was significantly low-er in research group than in control group [(56.19±37.87) vs. (152.54±53.92), P<0.01]. At PND70 (adulthood), the expres-sion of NR1, NR2A in the hippocampus was significantly higher in research group than in control group [NR1:(149.55%± 27.00%) vs. (100.00%±32.08%);NR2A:(171.54%±19.85%) vs. (100.00%±51.04%). P<0.05]. Conclusion Neonatal re-peated treatment of MK-801 increases the expression of NMDA receptor subunits NR1, NR2A in the hippocampus in adulthood while decreases the expression of NGF in the prefrontal cortex in adolescence, suggesting that neonatal block-ade of the NMDA receptor may influence the growth and development of the nervous system.
4.Saffold virus detection and epidemiological characteristics in children with acute respiratory infection or digestive tract infection in Tianjin
Wei WANG ; Jingfu HUANG ; Shuxiang LIN ; Ping SI ; Shengying LI ; Yimin ZHU ; Hongjiang YANG
Tianjin Medical Journal 2016;44(9):1164-1168
Objective To investigate prevalence and epidemiologic features of Saffold virus (SAFV) in hospitalized children with acute respiratory infection or digestive tract infection Tianjin area. Methods Nasopharyngeal aspirates from children with acute respiratory infection and fecal samples from children with digestive tract infection in Tianjin Children ’s Hospital were collected from January 2013 to December 2013. Viral nucleic acid was extracted, and SAFV infection was determined by using real-time quantitative PCR. Positive PCR products were sequenced. The sequencing results were aligned with known gene sequences of SAFV sequences in GenBank. The positive viral infection rate of nasopharyngeal aspirates and fecal samples, viral positive constituent ratio and positive detection rate in different age groups, seasonal distribution of SAFV infection were calculated. Other common respiratory tract or digestive tract viruses were also detected. Results Fourty-three (11.9%) nasopharyngeal aspirates from children with acute respiratory infection tested positive for SAFV. There was no significant difference between male and female infected children (aged between 6 d and 12 years old). The 79%(34/43) of the patients with SAFV infection aged under 1 year old. The infection most occurred in summer and winter. The 63 (16.4%) fecal samples from children with digestive tract infection tested positive for SAFV. There was significant difference between male and female infected children (aged between 5 h and 11 years old). SAFV infection was found to be year round. There was no significant difference in different age groups of nasopharyngeal aspirates and fecal samples. The mixed infection rate with SAFV and other respiratory tract or digestive tract viruses were 7.0%(3/43)and 12.7%(8/63), respectively. Conclusion Infection of SAFV had occurred in children with acute respiratory infection or digestive tract infection in Tianjin. SAFV has high detection rate in these children and is more common in children
aged under 1 year old. The data suggest that some of acute respiratory infection or digestive tract infections in pediatric patients are related to SAFV. The Clinical doctors should pay attention to them .
5.The experimental study of histology and apoptosis after the bone cement leakage into the intervertebral disc
Hui ZHAO ; Cai-Fang NI ; Long CHEN ; Tian-Si TANG ; Hui-Lin YANG ; Jian-Fei HUANG ;
Chinese Journal of Radiology 2000;0(12):-
Objective To discuss the histology change,apoptosis state and Bcl-2,Bax expression after the bone cement leakage into the intervertebral disc in vertebroplasty.Methods Eight majority canis familiaris were studied.Three lumbar intervertebral discs(L2 to L5)in each dog were randomly classified into three groups(control group,PMMA group,and CPC group),the canine discs were stabbed by 18-gauge needle,and 0.1 ml cement was injected into them.Control discs were only stabbed and injected with nothing.Histology of all discs was studied 24 weeks after the operation.Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and immunohistochemisty were used to detect apoptosis and Bcl-2,Bax expression in the discs.The data were statistically analyzed by SPSS 12.0.Results Intervertebral disc degeneration was not found in control groups.In bone cement groups,however,ruptured or serpentine patterned fibers,decreased cellularity of the nucleus pulposus and condensed matrix of the nucleus pulposus were found in histologic results.The Bax protein decreased in the order of control group, CPC group,and PMMA group.However,the Bcl-2 protein increased in the order of control group,CPC group,and PMMA group.The histology grade was significantly different among the three groups under ANOVA analysis(P
6.Clinical grade of generation of dendritic cells for immunotherapy.
Duozhuang, TANG ; Si, TAO ; Yang, CAO ; Jianfeng, ZHOU ; Ding, MA ; Wei, HUANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(3):265-8
In order to develop a protocol for clinical grade generation of dendritic cells (DCs) for cancer immunotherapy, aphereses were performed with the continuous flow cell separator and materials were derived from 10 leukemia patients that had achieved complete remission. Peripheral blood monocytes were cultured in vitro with GM-CSF, IL-4 for 6 days, then TNF-(the TNF-group) or TNF-, IL-1, IL-6, PGE2 (the cytokine mixture group) were added to promote maturation. Cell number was counted by hematology analyzer, and phenotype study (CD1a, CD14, CD83) was carried out by flow cytometry, and the function of DCs was examined by mixed lymphocyte reaction. The results showed that (0.70+/-0.13)x10(7)/mL (the TNF-alpha group) and (0.79+/-0.04)x10(7)/mL (the cytokine mixture group) DCs were generated respectively in peripheral blood obtained by leucapheresis. The phenotypes were as follows: CD1a+ (74.65+/-4.45)%, CD83+ (39.50+/-4.16)%, CD14+ (2.90+/-1.76)% in TNF-alpha group, and CD1a+ (81.86+/-5.87)%, CD83+ (81.65+/-6.36)%, CD14+ (2.46+/-1.68)% in the cytokine mixture group. It was concluded that leucapheresis may be a feasible way to provide large number of peripheral blood monocytes for DC generation, and combined administration of TNF-, IL-1, IL-6, and PGE2 may greatly promote maturity.
7.Expression of osteopontin and matrix metalloproteinase-9 in human malignant melanoma
Chang-Zheng HUANG ; Jing YANG ; Juan TAO ; Si-Yuan CHEN ; Ya-Ting TU ;
Chinese Journal of Dermatology 1994;0(06):-
Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P<0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P>0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.
8.Diagnosis and microsurgical treatment of intramedullary hemangioblastoma in cervical spinal cord
Xu-Hui HUI ; Chao YOU ; Rui-Chun LI ; Si-Qing HUANG ; Kai-Yong YANG ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To investigate the diagnosis and the microsurgical treatment of intramedullary hemangioblastoma in cervical spinal cord.Methods The signs of MRI,and the results of operations were analysed in 26 patients with the tumors.Rusults The tumors can be classified into two types:Solid type (14 cases)and cystic type(12 eases).All the tumors underwent total removal and were all hemangioblastoma confirmed by histopathologic examinations.Postoperatively,neurological status were improved in 17 patients, remained in 7 cases and worse in 2 cases.Conclusion For intrameduUary hemangioblastoma of cervical spinal cord MRI is of significant importance in the diagnosis of localization and the nature of the tumors which is conductive to selecting appropriate operative methods.There is high risk in operating at cervical section,but microsurgical total resection is the optimal method to stop the development of the clinical presentation.Opera- tive methods varied with the different typer of the tumor.It is the most important principal that dissection is performed along the correct interface and the tumor should be removed en bloc after it is devascularized.
9.Clinical Application Analysis of Andrographolide Total Ester Sulfonate Injection, a Traditional Chinese Medicine Licensed in China
ZHAO YING ; HUANG PU ; CHEN ZHE ; ZHENG SI-WEI ; YU JIN-YANG ; SHI CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(2):293-299
Andrographolide total ester sulfonate (ATES) injection is one of the products of traditional Chinese medicine (TCM) currently used against viral infection in China.ATES injection was approved for manufacturing and marketing in January 2002.It is indicated for acute respiratory infections,tonsillitis,chronic obstructive pulmonary disease,influenza,foot and mouth disease,bronchiolitis,herpangina,mumps,infectious mononucleosis and psychosis.However,its usage also carries risk.We investigated the use of ATES at the Wuhan Union Hospital from January 2014 to December 2014 and evaluated its real-word clinical application using the hospital centralized monitoring method.A total of 848 cases were enrolled in this study.In these cases,it was mainly used for postoperative anti-inflammation and treating upper respiratory infection,pneumonia and bronchitis.Among them,39.86% were contraindicated.Irregular medication of adults and children accounted for 1.91% and 23.38%,respectively.Improper choice of solvent accounted for 3.18%.The choice of intravenous drip versus aerosol inhalation was reasonable.A case of adverse events (AEs) was observed in the monitoring period,and the incidence of adverse drug reaction (ADR) of ATES injection was 0.12%.ATES injection in our hospital is relatively safe with a low incidence of adverse reactions.The study assesses the clinical usage and adverse reactions of ATES injection,and provides suggestions for rational use in clinical practice.
10.Effects of miR-20a-5p targeting KDM6B on the proliferation, migration and invasion of osteosarcoma cells
Bingliang LI ; Ya YANG ; Yingli HUANG ; Wen SI ; Xingwei LI ; Yuanmin ZHANG ; Jichao BIAN ; Yu CHEN
Journal of International Oncology 2021;48(2):65-73
Objective:To investigate the expressions of miR-20a-5p and lysine (K) demethylase 6B (KDM6B) in osteosarcoma tissues and the effects of miR-20a-5p targeting KDM6B on the proliferation, migration and invasion of osteosarcoma cells and tumor growth.Methods:The clinicopathological and paracancerous tissues of 20 patients with osteosarcoma admitted to the First Affiliated Hospital of Chinese Medical University from January 2017 to March 2019 were collected. Quantitative real-time PCR (qRT-PCR) was used to detect the expression levels of miR-20a-5p and KDM6B mRNA in tissues. The osteosarcoma MG63 cells were divided into control group, mimic NC group, miR-20a-5p mimic group, and NC+ empty vector group, miR-20a-5p+ empty vector group, miR-20a-5p+ KDM6B group. The expression levels of miR-20a-5p and KDM6B mRNA of all groups were detected by qRT-PCR. Western blotting was used to detect the expression level of KDM6B. CCK-8 assay, cell scratch test and Transwell test were used to detect cell proliferation, migration and invasion ability. According to the random number table method, nude mice were divided into NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group, with 5 mice in each group. Tumor growth ability was detected by tumor xenograft nude mouse models.Results:The relative expression level of miR-20a-5p mRNA in osteosarcoma tissues was 0.55±0.27, and that in paracancerous tissues was 1.22±0.28, with a statistically significant difference ( t=7.701, P<0.001). The relative expression level of KDM6B mRNA in osteosarcoma tissues was 1.66±0.19, and that in paracancerous tissues was 1.00±0.15, with a statistically significant difference ( t=12.219, P<0.001). After transfection of miR-20a-5p, KDM6B mRNA and protein expression levels decreased with the increase of miR-20a-5p expression level. After miR-20a-5p transfection for 48 h, the cell proliferation abilities of the blank control group, mimic NC group and miR-20a-5p mimic group were 0.83±0.04, 0.81±0.03 and 0.52±0.01 ( F=89.655, P<0.001), compared with the blank control group and mimic NC group, the cell proliferation ability was significantly inhibited in the miR-20a-5p mimic group (both P<0.001). The cell proliferation abilities of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were 0.83±0.05, 0.52±0.01 and 0.67±0.05 ( F=43.919, P<0.001), compared with the NC+ empty vector group, the cell proliferation ability was significantly inhibited in the miR-20a-5p+ empty vector group ( P<0.001); compared with the miR-20a-5p+ empty vector group, the cell proliferation ability of miR-20a-5p+ KDM6B group increased significantly ( P<0.001). The scratch healing rates of the blank control group, mimic NC group and miR-20a-5p mimic group were (32.51±2.73)%, (30.26±3.22)% and (13.52±1.77)% ( F=46.314, P<0.001), compared with the control group and the mimic NC group, the scratch healing rate of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The scratch healing rates of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (31.34±3.11)%, (12.15±1.64)% and (28.93±2.89)% ( F=47.511, P<0.001), compared with the NC+ empty vector group, the scratch healing rate of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the scratch healing rate of miR-20a-5p+ KDM6B group was significantly increased ( P=0.001). The numbers of transmembrane cells in the blank control group, mimic NC group and miR-20a-5p mimic group were 114±16, 108±11 and 42±6 ( F=36.282, P<0.001), compared with the control group and mimic NC group, the number of transmembrane cells of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The numbers of transmembrane cells in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group was 143±11, 39±4 and 139±12 ( F=112.120, P<0.001), compared with the NC+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ KDM6B group was increased significantly ( P<0.001). The tumor volumes of mice for 21 d in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (1 667.50±250.40) mm 3, (129.20±21.00) mm 3 and (775.41±77.51) mm 3 respectively, with a statistically significant difference ( F=77.651, P<0.001). The tumor weights of the 3 groups were (1.35±0.18) g, (0.12±0.01) g and (0.61±0.03) g respectively, with a statistically significant difference ( F=104.191, P<0.001). Conclusion:The expression of miR-20a-5p is significantly decreased in osteosarcoma tissues, and the expression of KDM6B is significantly increased in osteosarcoma tissues. Overexpression of miR-20a-5p may inhibit the proliferation, migration and invasion of osteosarcoma cells and tumor growth by targeting to reduce the expression of KDM6B.