1.Effect of Buyang Huanwu Decoction on Neural Cell Apoptosis After Cerebral Ischemia in Gerbils and Its Mechanism
Zhilong LIU ; Hanping SONG ; Changqing DENG ; Shaojua HAN
Journal of Guangzhou University of Traditional Chinese Medicine 2000;0(04):-
To study the effect of Buyang Huanwu Decoction (BHD) on neural cell apoptosis after cerebral ischemia in gerbils and its mechanism, gerbil model with cerebral ischemia was established by clamping of bilateral carotid arteries and then reperfusion. Neural cell apoptosis of hippocampal gyrus slides was observed under electron microscope 120 hours after reperfusion. The activities of Na + -K + ATPase , Ca 2+ ATPase , Mg 2+ ATPase and NOS, and the contents of lactic acid, NO and amino acid in cerebral tissues were detected 120 hours after reperfusion. The results showed that BHD can reduce the incidence of neural cell apoptosis of hippocampal gyrus, prevent the decrease of Na + -K + ATPase and Ca 2+ ATPase activities, increase NO content and NOS activity, and reduce cerebral glucose content. It is indicated that BHD can counteract neural cell apoptosis after cerebral ischemia in gerbils and its mechanism may be related to the improvement of cerebral energy metabolism, regulation of NO synthesis and antagonism of toxic effect of excitatory amino acid.
2.Analysis of 655 cases of mycoplasma pneumoniae infection in children
Lian YE ; Yibin DENG ; Fang WANG ; Kun FANG ; Hua GOU ; Zhilong CAI
International Journal of Laboratory Medicine 2015;(12):1676-1677,1679
Objective To analyze the MP infection status in children with respiratory tract disease and the correlation with gen‐der ,age ,season and clinical conditions .Methods To investigate the clinical data retrospectively of 655 children with respiratory tract infection from January to December 2013 .Results The positive rate of MP antibody was 48 .09% with a higher incidence in girls than boys ,and those above 3 were more susceptible to it .Winter and spring were the peak seasons .The older group had a higher positive rate of MP antibody ,together with high morbidity of MPP and LP ,the differences were statistically significant (P<0 .05) ,while the younger group was inclined to get higher WBC count and percentage of increasing CRP and neutrophils ,the differ‐ences were statistically significant(P<0 .05) .Conclusion MP is a common pathogenic bacteria causes respiratory tract infection in children above 3 years ,especially in winter and spring .The antibody positive rate rise with age and the infected children are more likely to have pneumonia meanwhile the younger group has higher WBC count ,in which more cases get higher level of CRP and neutrophils .
3.Treatment of multiple level noncontiguous thoracolumbar fractures with posterior pedicle screw
Biao YANG ; Wei JIN ; Ansong PING ; Lin CAI ; Zhilong CHEN ; Zhouming DENG ; Renxiong WEI ; Bin REN
Clinical Medicine of China 2011;27(12):1305-1308
Objective To explore the clinical effect of posterior pedicle screw internal fixation in the treatment of multiple level noncontiguous thoracolumbar fractures.Methods Thirteen patients with multiple level noncontiguous thoracolumbar fractures were treated by posterior pedicle screw internal fixation.The Frankel score,percentage of vertebral compression and Cobb angle of the injured vertebral segment were analyzed to evaluate the surgery efficacy.Results All patients were followed up from 12 to 24 months ( averaged 15 months ).All cases achieved bone fusion with no implant failure.The Cobb angle of the injured vertebral segment was corrected from preoperative(22.2 ±5.3) degree to postoperative(5.3 ±3.5) degree and(6.2 ±3.6) degree at the last follow up.The percentage of vertebral compression was corrected from preoperative (45.7 ± 14.1 )% to postoperative ( 6.1 ± 3.8 ) % and ( 7.2 ± 3.9 ) % at the last follow up.All improvements showed significant differences when compared statistically( t =15.03,t =12.05,Ps <0.05 ).The spinal cord function was improved 1 to 2 degree in all patients except 2 patients of grade A.Conclusion The posterior fixation with pedicle screw is a secure,safe and effective method in treating multiple level noncontiguous thoracolumbar fracture.
4.Application of Q self-traction method in endoscopic submucosal dissection to the treatment of large early esophageal cancer (with video)
Zhilong CHEN ; Xiaolu LIN ; Wanyin DENG ; Jinhui ZHENG ; Xianbin GUO ; Guowei WANG ; Haining LIN ; Wei LIANG
Chinese Journal of Digestive Endoscopy 2020;37(3):163-168
Objective:To preliminarily study the effect of Q self-traction endoscopic submucosal dissection (Q-ESD) on treatment of large early esophageal cancer (EEC).Methods:A retrospective analysis was performed on the data of 82 cases of large EEC (single lesion>1/2 cross-section diameter or longitudinal diameter length >5 cm) who underwent ESD on Fujian Provincial Hospital between January 2015 and December 2018. According to the treatment schedule, patients were divided into the conventional ESD group (n=44) and the Q-ESD group (n=38). The procedural area, time, and speed, en bloc resection rate, complete resection rate and complications of the two groups were analyzed.Results:All of the 82 lesions were resected completely under endoscope. There was no statistical difference in the procedural area [779.8 (329.9-2 552.5)mm 2 VS 875.7 (417.8-1 914.8)mm 2, U=155, P=0.636], procedural time [63 (41-177)min VS 59 (42-169)min, U=171, P=0.167] and complete resection rate [94.7% (36/38) VS 93.2% (41/44), χ2=0.086, P=0.769] between the Q-ESD group and the conventional ESD group. Compared with the conventional ESD group, the Q-ESD group had a faster dissection speed [14.9 (5.4-20.8) mm 2/min VS 9.0 (5.0-19.5) mm 2/min, U=142, P=0.035], lower muscularis propria injury rate [7.9% (3/38) VS 27.3% (12/44), χ2=5.123, P=0.023], and a lower stricture rate [5.3% (2/38) VS 20.5% (9/44), χ2=4.051, P=0.044]. No other adverse events occurred except for one case of perforation in the conventional ESD group. Conclusion:The new traction technique of Q-ESD is a safe and effective treatment for large EEC.
5.lncRNA THAP7-AS1 affects the glycolysis of gastric cancer cells by regulating METTL3 mediated m6A modification
Zhilong DENG ; Xianmo YANG ; Can WANG ; Xian SU ; Lingfei GUO
Tumor 2023;43(10):781-798
Objective:To explore the effect of long noncoding RNA(lncRNA)THAP7-AS1 on the glycolysis of gastric cancer(GC)cells by regulating methyltransferase-like 3(METTL3)mediated N6-methyladenosine(m6A)modification. Methods:Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze the expression levels of THAP7-AS1 and METTL3 in GC tissues and their relationship with the overall survival of GC patients.Real-time fluorescence quantitative PCR and Western blotting were used to analyze the expression of THAP7-AS1,METTL3 mRNA,glucose transporter 1(GLUT1)mRNA,and METTL3 protein in GC tissues and paracancerous tissues samples collected from 80 GC patients in Department of Oncology,The Third Affiliated Hospital of Zunyi Medical University(the First People's Hospital of Zunyi),and the relationship between THAP7-AS1 levels and the clinicopathological characteristics of GC patients was analyzed.Real-time fluorescence quantitative PCR and Western blotting were used to verify the expression of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA,and METTL3 protein in GES-1,BGC-823 and SGC-7901 cells.Lentiviral infection was used to knock-down THAP7-AS1 or overexpress METTL3 BGC-823 and SGC-7901 cells,and real-time fluorescence quantitative PCR and Western blotting were used to examine effect of different treatment on the expression of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA,and METTL3 protein;colorimetry assay was used to detect the m6A modification level in the total RNA;methylated RNA immunoprecipitation(MeRIP)-quantitative PCR(qPCR)was used to detect the GLUT1 m6A modification level;glycolysis stress test kits were used to detect the extracellular acidification rate(ECAR),glucose uptake and lactate production of treated GC cells;Western blotting was used to examine the expression levels of METTL3,GLUT1,M2 type pyruvate kinase(PKM2)and lactic dehydrogenase(LDHA)proteins in treated GC cells;EdU staining,wound healing assay and Transwell invasion assay were used to evaluate the proliferation,migration and invasion of treated GC cells.Finally,a mouse model of subcutaneously transplanted GC tumor was established using nude mice,and the effect of knocking-down THAP7-AS1 was assessed by measuring the tumor volume and weight,as well as the expression levels of METTL3 and GLUT1 proteins in transplanted GC tumor tissues. Results:Analysis of the GEPIA database showed that the expression levels of THAP7-AS1 and METTL3 was higher in GC tissue than those in normal gastric tissues,and the expression levels of THAP7-AS1 and METTL3 are negatively correlated with overall survival of GC patients(P<0.05).Compared with the paracancerous tissues(or normal gastric epithelial cells),the expression levels of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA and METTL3 protein was significantly increased in GC tissues(or GC cells),and the higher the expression of THAP7-AS1,the higher the TNM stage,the lower the degree of tumor differentiation,and the easier the occurrence of microvascular infiltration and lymph node metastasis(P<0.05).Knocking-down of THAP7-AS1 down-regulated the expression levels of METTL3 mRNA,GLUT1 mRNA and METTL3 protein,the m6A modification levels in total RNA and GLUT1,the ECAR levels,the glucose uptake,the lactate production,EdU positive rate,scratch healing rate,the number of invaded cells,and the expression levels of glycolysis-related proteins(METTL3,GLUT1,PKM2 and LDHA)in GC cells(P<0.05).Overexpression of METTL3 could partially reverse these effects of THAP7-AS1 knock-down(P<0.05).In vivo experiments showed that THAP7-AS1 knock-down can obviously inhibit the growth of transplanted GC tumors(P<0.05). Conclusion:lncRNA THAP7-AS1 can promote the glycolysis which further promotes the proliferation,migration and invasion of GC cells by regulating METTL3 mediated m6A modification.
6.Identification and functional analysis of soybean stearoyl-ACP Δ⁹ desaturase (GmSAD) gene family.
Mimi DENG ; Baoling LIU ; Zhilong WANG ; Jin'ai XUE ; Hongmei ZHANG ; Runzhi LI
Chinese Journal of Biotechnology 2020;36(4):716-731
Stearoyl-ACP Δ⁹ desaturase (SAD) catalyzes the synthesis of monounsaturated oleic acid or palmitoleic acid in plastids. SAD is the key enzyme to control the ratio of saturated fatty acids to unsaturated fatty acids in plant cells. In order to analyze the regulation mechanism of soybean oleic acid synthesis, soybean (Glycine max) GmSAD family members were genome-wide identified, and their conserved functional domains and physicochemical properties were also analyzed by bioinformatics tools. The spatiotemporal expression profile of each member of GmSADs was detected by qRT-PCR. The expression vectors of GmSAD5 were constructed. The enzyme activity and biological function of GmSAD5 were examined by Agrobacterium-mediated transient expression in Nicotiana tabacum leaves and genetic transformation of oleic acid-deficient yeast (Saccharomyces cerevisiae) mutant BY4389. Results show that the soybean genome contains five GmSAD family members, all encoding an enzyme protein with diiron center and two conservative histidine enrichment motifs (EENRHG and DEKRHE) specific to SAD enzymes. The active enzyme protein was predicted as a homodimer. Phylogenetic analysis indicated that five GmSADs were divided into two subgroups, which were closely related to AtSSI2 and AtSAD6, respectively. The expression profiles of GmSAD members were significantly different in soybean roots, stems, leaves, flowers, and seeds at different developmental stages. Among them, GmSAD5 expressed highly in the middle and late stages of developmental seeds, which coincided with the oil accumulation period. Transient expression of GmSAD5 in tobacco leaves increased the oleic acid and total oil content in leaf tissue by 5.56% and 2.73%, respectively, while stearic acid content was reduced by 2.46%. Functional complementation assay in defective yeast strain BY4389 demonstrated that overexpression of GmSAD5 was able to restore the synthesis of monounsaturated oleic acid, resulting in high oil accumulation. Taken together, soybean GmSAD5 has strong selectivity to stearic acid substrates and can efficiently catalyze the biosynthesis of monounsaturated oleic acid. It lays the foundation for the study of soybean seed oleic acid and total oil accumulation mechanism, providing an excellent target for genetic improvement of oil quality in soybean.
Fatty Acid Desaturases
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genetics
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metabolism
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Gene Expression Profiling
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Oleic Acid
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biosynthesis
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Phylogeny
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Plant Proteins
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genetics
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Seeds
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chemistry
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Soybeans
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classification
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enzymology
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genetics