1.Study on the Role and Mechanism of METTL3 Mediating the Up-regulation of m6A Modified Long Non-coding RNA THAP7-AS1 in Promoting the Occurrence of Lung Cancer.
Yu ZHANG ; Yanhong WANG ; Mei LIU
Chinese Journal of Lung Cancer 2024;26(12):919-933
BACKGROUND:
Lung cancer is a major threat to human health. The molecular mechanisms related to the occurrence and development of lung cancer are complex and poorly known. Exploring molecular markers related to the development of lung cancer is helpful to improve the effect of early diagnosis and treatment. Long non-coding RNA (lncRNA) THAP7-AS1 is known to be highly expressed in gastric cancer, but has been less studied in other cancers. The aim of the study is to explore the role and mechanism of methyltransferase-like 3 (METTL3) mediated up-regulation of N6-methyladenosine (m6A) modified lncRNA THAP7-AS1 expression in promoting the development of lung cancer.
METHODS:
Samples of 120 lung cancer and corresponding paracancerous tissues were collected. LncRNA microarrays were used to analyze differentially expressed lncRNAs. THAP7-AS1 levels were detected in lung cancer, adjacent normal tissues and lung cancer cell lines by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The diagnostic value of THAP7-AS1 in lung cancer and the relationship between THAP7-AS1 expression and survival rate and clinicopathological parameters were analyzed. Bioinformatics analysis, methylated RNA immunoprecipitation (meRIP), RNA pull-down and RNA-immunoprecipitation (RIP) assay were used to investigate the molecular regulation mechanism of THAP7-AS1. Cell proliferation, migration, invasion and tumorigenesis of SPC-A-1 and NCI-H1299 cells were determined by MTS, colony-formation, scratch, Transwell and xenotransplantation in vivo, respectively. Expression levels of phosphoinositide 3-kinase/protein kenase B (PI3K/AKT) signal pathway related protein were detected by Western blot.
RESULTS:
Expression levels of THAP7-AS1 were higher in lung cancer tissues and cell lines (P<0.05). THAP7-AS1 has certain diagnostic value in lung cancer [area under the curve (AUC)=0.737], and its expression associated with overall survival rate, tumor size, tumor-node-metastasis (TNM) stage and lymph node metastasis (P<0.05). METTL3-mediated m6A modification enhanced THAP7-AS1 expression. The cell proliferation, migration, invasion and the volume and mass of transplanted tumor were all higher in the THAP7-AS1 group compared with the NC group and sh-NC group of SPC-A-1 and NCI-H1299 cells, while the cell proliferation, migration and invasion were lower in the sh-THAP7-AS1 group (P<0.05). THAP7-AS1 binds specifically to Cullin 4B (CUL4B). The cell proliferation, migration, invasion, and expression levels of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphoinositide-3 kinase, catalytic subunit delta (PIK3CD), phospho-phosphatidylinositol 3-kinase (p-PI3K), phospho-protein kinase B (p-AKT) and phospho-mammalian target of rapamycin (p-mTOR) were higher in the THAP7-AS1 group compared with the Vector group of SPC-A-1 and NCI-H1299 cells (P<0.05).
CONCLUSIONS
LncRNA THAP7-AS1 is stably expressed through m6A modification mediated by METTL3, and combines with CUL4B to activate PI3K/AKT signal pathway, which promotes the occurrence and development of lung cancer.
Humans
;
Lung Neoplasms/pathology*
;
RNA, Long Noncoding/metabolism*
;
Phosphatidylinositol 3-Kinases/metabolism*
;
Up-Regulation
;
Proto-Oncogene Proteins c-akt/metabolism*
;
Cell Line, Tumor
;
Cell Proliferation/genetics*
;
Gene Expression Regulation, Neoplastic
;
Methyltransferases/metabolism*
;
Cullin Proteins/genetics*
2. Effect of long-term and short-term mild hypothermia in severe traumatic brain injury: a comparative study
Zhigang LI ; Xirong JI ; Taizu ZHENG ; Yinjun ZHANG ; Zhenchen LI
Chinese Critical Care Medicine 2019;31(12):1440-1444
Objective:
To explore the effects of different mild hypothermia therapy time on the efficacy and complications of patients with severe traumatic brain injury (STBI).
Methods:
A retrospective research method was used. 132 patients with STBI given mild hypothermia therapy admitted to the Third Affiliated Hospital of Gansu University of Traditional Chinese Medicine from January 2010 to December 2018 were enrolled. According to the days of mild hypothermia therapy, the patients were divided into 2-day mild hypothermia treatment group, 5-day mild hypothermia treatment group and 14-day mild hypothermia treatment group. Glasgow coma score (GCS) after treatment of 10 days and 30 days, 30-day disability rate and mortality, coma time, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), D-dimer, and the incidence of complications such as decreased blood pressure, decreased gastrointestinal motility, infection, nonunion of scalp, cerebrospinal fluid leakage, etc. were compared among three groups.
Results:
Among the 132 patients with STBI, there were 44 cases in the 2-day mild hypothermia treatment group, 45 cases in 5-day mild hypothermia treatment group, and 43 cases in 14-day mild hypothermia treatment group. There was no significant difference in gender, age, GCS score before treatment or time from injury to admission among three groups. Compared with 2-day mild hypothermia treatment group, the GCS score 10 days and 30 days after treatment in 5-day mild hypothermia treatment group and 14-day mild hypothermia treatment group were significantly higher (11.61±2.23, 10.17±2.03 vs. 6.79±1.49; 13.15±2.53, 11.24±2.24 vs. 8.79±1.59), the coma time were shorten (days: 5.79±1.89, 5.45±1.72 vs. 13.65±2.73), and 30-day disability rate and mortality were significantly decreased [13.33% (6/45), 11.63% (5/43) vs. 22.73% (10/44); 17.78% (8/45), 16.28% (7/43) vs. 31.82% (14/44)], PT and APTT were reduced obviously (s: 20.14±4.12, 22.54±3.56 vs. 30.67±5.19; 35.14±12.41, 38.59±13.54 vs. 56.67±10.62), Fib rose obviously (g/L: 1.84±0.25, 1.98±0.27 vs. 0.67±0.12), and D-dimer reduced obviously (mg/L: 53.10±19.84, 49.20±20.13 vs. 102.60±20.13), with statistically significant differences (all
3.Monitoring and analysis of food microbial contamination in rural compulsory education schools in Baiyin City from 2013 to 2021
YANG Junpeng, CHEN Anming, ZHANG Guoqin, ZENG Tongxia, QIANG Weidong
Chinese Journal of School Health 2022;43(6):934-936
Objective:
To analyze the level and trend of microbial contamination in food in rural compulsory education schools in Baiyin City, so as to provide a reference for playing an early warning role in eliminating potential food safety hazards as soon as possible.
Methods:
According to the "National Food Contaminants and Harmful Factors Risk Monitoring Workbook", etc., 8 pathogenic bacteria such as Salmonella, Listeria monocytogenes, Staphylococcus aureus, total bacterial count, and 2 hygienic indicator bacteria of coliform were tested.
Results:
A total of 908 food samples were collected from 2013 to 2021, 47 were positive and the overall positive rate was 5.18%. The positive rate of samples collected in different years showed a downward trend year by year. The detected pathogenic bacteria ranked from high to low as follows: Escherichia coli (1.87%), Bacillus cereus (0.99%), Staphylococcus aureus (0.44%), mold (0.33%), Listeria monocytogenes (0.22%). Hygiene indicator bacteria total number of bacteria exceeded the standard rate was higher (2.09%). Among all kinds of food, milk and dairy products, frozen food, cold food, barbecue, rice and flour products had higher positive rates. According to quarterly statistics, the positive rate in the first and second quarters was relatively high.
Conclusion
There are different degrees of microbial contamination risks in the food of rural compulsory education schools. All relevant departments should pay attention to the current situation of school food hygiene and safety in poverty stricken areas, and strengthen rural compulsory education school food, especially for milk and dairy products, frozen food, cold food, Supervision of key foods such as barbecue, rice and flour products, and efforts to improve their safety standards, do a good job in school food hygiene and safety, and ensure the healthy growth of young people.
4.Expression of BLM mRNA in leukemia and its clinical significance
Yongjuan ZHANG ; Xiaodong HE ; Yujie SUN ; Baiyin ZHANG ; Rong HUANG ; Jinling JI ; Zuojun SHEN
Chinese Journal of Laboratory Medicine 2012;(12):1130-1134
Objective To investigate the correlation between the expression level of BLM and its clinical significance in leukemia.Methods 125 bone marrow specimens of inpatients and outpatients with leukemia were collected in Anhui provincial hospital from January 2011 to December 2011.125 leukemia patients were diagnosed and classified into acute leukemia (AL,n =66) and chronic myelogenous leukemia (CML,n =59) by Morphologic and Immunologic criteria,5 non-tumor individuals were included as control group.The BLM mRNA expressions were by reverse transcription-polymerase chain reaction(RT-PCR).The specimens were devided into groups according to the age,gender,leukemia type,peripheral blood leukocyte counts,hepatomegalia and(or) splenomegaly,fusion gene,chromosome karyotype,whether first visit and transplantation.The expression of BLM gene in each group and the correlation with above factors were retrospectively analysed.The statistical methods such as chi-square test,single factor variance analysis,t test and Pearson correlation test were mainly used.Results BLM mRNA was detected in leukemia.In bone marrow cells,the BLM gene expression was positive in 71 patients and negative in 54 patients.But none of 5 non-tumor bone marrow cells expressed BLM gene.The difference of BLM expression between patients and controls was statistically significant in two groups,i.e.peripheral blood leukocyte counts and fusion gene (x2 =14.730,22.399 ; P < 0.05),but there is no statistical significant differences in other groups.The expressions of BLM mRNA in leukemia patients who had been treated with chemotherapy were lower than those newly diagnosed (0.1788 ± 0.1091 vs 0.3276 ± 0.2016 ; P < 0.05).Moreover,BLM mRNA level in post-bone marrow transplant patients was lower than those not treated (0.1271 ± 0.1009 vs 0.2902 ±0.2034 ; P < 0.05).Pearson correlation analysis showed that higher BLM mRNA expression positively correlated with fusion gene (r =0.357,P < 0.01) and chromosome abnormality (r =0.279,P < 0.05).Conclusion The BLM mRNA expression level of measurement can be used as judgment for leukemia patients disease severity and the index of prognosis,testing the level may provide a basis for clinical and curative effect judgment.
5.The relationship between IgA nephropathy and low molecular weight urinary protein
Baiyin QIAN ; Hongyun LU ; Shuang LI ; Hua ZHANG ; Zhonghe LI ; Hequn ZOU
Chongqing Medicine 2014;(32):4291-4293
Objective To investigate the relationship among urinary protein molecular weight of IgA nephropathy ,renal tubu‐lointerstitium damage and clinical index ,and discuss the role of low molecular weight urinary protein in the mechanism of IgA pro‐gression .Methods A total of 34 patients with biopsy proven IgAN were studied .We detect the molecular weight of urinary protein by SDS‐PAGE .Data were processed with the classes of tubulointerstitial lesions and laboratory tests such as 24h urinary protein quantitation and serum creatinine(Scr) .Results The urinary protein were divided into four types according the SDS‐PAGE:the ball urinary protein group ,physiologic urinary protein group ,23 × 103 proteinuria group and 10 × 103 proteinuria group .Scores of tubu‐lointerstitial lesions ,twenty four hour urine protein quantitation and Scr level were all significantly higher in 10 × 103 proteinuria group than 23 × 103 proteinuria group (P<0 .05) .Conclusion 10 × 103 proteinuria may have close relationship with the progression of IgAN ,and it may be a useful index for the degree of tubulointerstitial lesions of IgAN .
6.Iatrogenic lipogranuloma of the penis.
Zheng-Chao ZHANG ; Tong-Wei ZENG ; Jun-Ping HAN ; Yong-Qiang XIE
Asian Journal of Andrology 2023;25(1):147-148
Male
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Humans
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Penis
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Granuloma/etiology*
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Pelvis
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Iatrogenic Disease
7.Development of a real-time fluorescence quantitative PCR method for detection of FPGS mRNA expression in methotrexate enantiomer-resistant A549 cell lines and patients with leukemia
Li SUN ; Xiaodong HE ; Yujie SUN ; Weidong XU ; Daojing LI ; Baiyin ZHANG ; Yongjuan ZHANG ; Rui LIU ; Zuojun SHEN
Chinese Journal of Laboratory Medicine 2011;34(8):722-726
Objective To establish a real-time fluorescence quantitative PCR method for detection of the different expression level of FPGS in methotrexate enantiomer-resistant A549 cell lines,and observe FPGS mRNA expression in patients with leukemia.Methods A real-time fluorescence quantitative PCR method for detection FPGS mRNA was established using SYBR Green Ⅰ as fluorescence and β-actin as reference.The method was evaluated by Ct,correlation coefficient,slope,repeatability curve,melting curve and amplification efficiency curve.The expression levels of FPGS gene in methotrexate enantiomer-resistant A549 cell lines and methotrexate resistant leukemia cells in bone marrow were detected by the method.Results The standard curves had a high linear relationship between cycle threshold and template concentration.The correlation coefficients of FPGS and β-actin were 0.996 8 and 0.998 7,and the slopes were -3.595 and -3.740,respectively.The inter-coefficient of variation was from 1.27% to 2.95%.The intra-coefficient of variation was 3.82%.The method was characterized with specific melting curve and similar amplification efficiency(slope was 0.021 7).The relative contents of FPGS mRNA were(3.51 ±0.66),(0.16 ±0.01) and(1.00 ±0.31) in L-(+)-MTX/A549 cells(L),D-(-)-MTX/A549 cells(D)and A549 parent cells,and there was statistically difference among the three groups(F = 64.45 ,P< 0.01)Statistical difference was observed between L and D(q =9.29,P<0.01).After treated with MTX,the expression level of FPGS mRNA was(0.35 ± 0.04) in methotrexate resistant leukemia patients,compared with(1.00 ± 0.44) before treatment.Statistical difference was observed(t = 8.83 ,P< 0.01).Conclusions The real-time fluorescence quantitative PCR is suitable for the quantification of FPGS.The expression levels of FPGS in methotrexate resistant leukemia cells in bone marrow and drug resistant cells are different.Two enantiomer forms of methotrexate may play different roles in drug resistance mechanisms.
8.Brain protective effect of ginkgo biloba extract on patients with meningioma resection
Botao CAI ; Cunyan LI ; Zhiying DENG ; Jiwei TANG ; Baiyin ZHANG
Journal of Chinese Physician 2019;21(4):516-520
Objective To investigate the effects of ginkgo biloba extract on the early postoperative cognitive function and the serum neuron specific enolase (NSE),S100β,tumor necrosis factor-α (TNF-α),interleukin (IL)-6,superoxide dismutase (SOD) and malondialdehyde (MDA) in patients with meningioma resection.Methods 60 patients with meningioma scheduled for elective intracranial tumor resection were enrolled and randomly divided into two groups (n =30 each):ginkgo biloba extract group (group G) and control group (group C).Ginkgo biloba extract was infused intravenously 30 min before anesthesia induction in group G,while group C received the equal volume of normal saline.Venous blood sample were taken at three time points:30 min before induction (T1),extubation (T2) and 24 h after operation (T3)for determination of serum concentration of NSE,S100β,TNF-α,IL-6,SOD and MDA.Cognitive function was evaluated at 1 d before and 3 d after surgery using Mini-mental State Examination (MMSE),Wechsler Adult Intelligence Scale (WAIS) and Wechsler Memory Scale (WMS).Results Compared with T1,the concentration of NSE,S100β,TNF-α,IL-6 and MDA in serum were significantly increased at T2 and T3 in the two groups,while SOD was decreased (P < 0.05).Compared with group C,the concentration of NSE,S100β,TNF-α,IL-6 and MDA were significantly decreased at T2 and T3 in group G,while SOD was increased (P < 0.05).Compared with 1 d before surgery,the scores of MMSE,digit accumulation,digit breadth (forward),vocabulary association and digit symbol replacement were significantly decreased and tracing connection time was significantly increased on the 3rd day after surgery in the two groups.The scores of digital breadth (reverse) and visual regeneration were significantly decreased in group C (P < 0.05).Compared with group C,the scores of digit accumulation,digit breadth (forward and reverse) and visual regeneration were significantly increased on the 3rd day after surgery in group G (P < 0.05).The incidence of postoperative cognitive dysfunction (POCD) at 3 d after surgery in group G was significantly lower than group C (P < 0.05).Conclusions The pretreatment of ginkgo biloba extract can decrease the incidence of early POCD in patients with meningioma resection,and its mechanism may be related to the reduction of inflammatory reaction and antioxidation.
9.Serologic characteristics and population distribution of subtypes B2 and AB2 of ABO blood group.
Fu-Cai DUAN ; Ming-Lu WANG ; Ke-Li ZHOU ; Da-Yuan LI ; Qin-Yong ZHANG ; Ai-Ping MA ; He-Ying YANG ; Jian-Hua LI ; Yuan-Yuan LIU ; Fang XIAO ; Ying-Xue GAO
Journal of Experimental Hematology 2010;18(5):1331-1334
This study was aimed to investigate the serologic characteristics, genetic background and population distribution of B2 and AB2 subtype in Chinese ABO blood group. The classic blood group serological technology was used to detect ABO blood group of the propositus and their family members, the anti-B1 serum prepared by yourself was used to investigate the distribution of B1/B2 and AB1/AB2 subtype of the blood donor. The results indicated that the antigen of propositus was AB2 subtype and that of his child was B2 subtype. The anti-B1 antibody was detected in blood serum of propositus; the antigen of 3 from 2318 blood donors with B blood group were found to be B2 subtype, the antigen of 2 from 826 blood donors with AB blood group were found to be AB2 subtype. The investigation on propositus and the 3 B2 blood donor families showed that B2 antigen displays genetic characteristics of blood group. It is concluded that B2/AB2 subtype is from family inheritance, while B2 subtype is amounted to 0.129% in B blood group, and AB2 subtype is amounted to 0.224% in AB blood group.
ABO Blood-Group System
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classification
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genetics
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immunology
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Blood Grouping and Crossmatching
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China
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Female
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Genetics, Population
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Humans
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Male
;
Middle Aged
10. Resource Investigation and Conservation of Medicinal Plants in Hasi Mountain Nature Reserve of Gansu Province
Qi-li ZHANG ; Lu-ming REN ; Ding HAN ; Xiao-rong ZHANG ; Fu-yun LI ; Zhi-jia CUI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(24):131-135
Objective:To explore the wild medicinal plant species and utilization of Hasi mountain nature reserve,in order to provide the references for reasonable utilization and protection of the medicinal plant resources in this area. Method:The survey was conducted based on the technical scheme of the Fourth National Survey on Chinese Material Medica Resources. By consulting literature,collecting medicinal plants specimen and visiting survey area,researchers collected and summarized the wild medicinal plant species, and analyzed the results of reserves. Result:The results showed 7 varieties in 247 kinds of wild medicinal plants in Hasi mountain nature reserve,which belonged to 161 genera in 61 families;by the medicinal parts of all wild medicinal plants,the whole plant of a total of 124 species could be used,accounting for 48.82% of the total species,by traditional Chinese medicine efficacy, antipyretic herbs were the majority,accounting for 32.68% of the total. In addition,there were 8 kinds of rare and endangered wild medicinal plants,such as Ephedra sinica,Gentiana dahurica and Epipactis helleborine. Conclusion:Hasi mountain nature reserve has rich wild medicinal plant resources,and is an important part of medicinal plant resources and protection areas of Jingyuan county and Loess Plateau. However, because they are serious affected by incontinent excavation and insect pest,efforts shall be made in scientific protection and rational development.