1.Effects of zinc finger transcription factor 580 in regulating autophagy and extracelluar matrix secretion of PANC1 cells
Baiqing LIU ; Wei XU ; Wencheng ZHANG ; Shihai XIA
Chinese Journal of Pancreatology 2025;25(5):355-360
Objective:To investigate the regulatory effect of zinc finger transcription factor 580 (ZNF580) gene on autophagy and extracellular matrix (ECM) secretion in human pancreatic cancer cells PANC1.Methods:PANC1 cells were transfected with 500 ng/ml short hairpin RNA-ZNF580 (shRNA-ZNF580) and a ZNF580 expression vector with a green fluorescent protein reporter gene (GFP-ZNF580) using lentiviral transfection to establish the ZNF580-silenced group and ZNF580-overexpression group, respectively. PANC1 cells were treated with 10 mmol/L rapamycin (RA), a cell autophagy inducer, and the autophagy inhibitor LY294002 for 2 hours to construct the autophagy-induced group and autophagy-inhibited group, respectively. The autophagy inhibition+ZNF580 silencing group was established by transfecting PANC1 cells with 500 ng/ml sh-ZNF580 using lentiviral transfection while simultaneously adding 10 mmol/L LY294002. PANC1 cells cultured in conventional medium served as control group. The expression levels of ZNF580 protein and autophagy-related proteins ATG7 and LC3 in PANC1 cells from each group were detected by Western blot. The expression changes of ECM secretion-related markers type I collagen (Col-Ⅰ), Col-Ⅲ, fibronectin (FN), and tumor necrosis factor-α (TNF-α) in PANC1 cells were measured by ELISA.Results:Compared with control group, the protein expression levels of ATG7, LC3-Ⅰ, and LC3-Ⅱ in PANC1 cells of the ZNF580-silenced group were significantly decreased (0.40±0.04 vs 0.81±0.13, 0.66±0.08 vs 2.0±0.45, 0.78±0.10 vs 1.89±0.23), while they were significantly increased in the ZNF580-overexpression group (2.07±0.17 vs 0.83±0.09, 1.21±0.37 vs 0.88±0.09, 0.77±0.16 vs 0.37±0.06). The protein expression level of ZNF580 in PANC1 cells of the autophagy inhibition group was significantly down-regulated compared with the control group (0.40±0.15 vs 1.07±0.18), while it was significantly up-regulated in the autophagy induction group (1.59±0.25 vs 0.67±0.09). Compared with the control group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells were significantly decreased in the ZNF580-silenced group (5.02±0.81 vs 8.38±0.83, 6.17±0.83 vs 10.73±1.69, 28.66±2.47 vs 45.20±4.31, 10.09±1.32 vs 19.48±2.77), which were significantly increased in the ZNF580-overexpression group (19.28±2.05 vs 8.38±0.83, 28.29±5.96 vs 10.73±1.69, 103.22±6.37 vs 45.20±4.31, 46.78±6.96 vs 19.48±2.77), significantly decreased in the autophagy inhibition group (5.10±0.66 vs 9.01±1.24, 7.22±0.67 vs 11.83±1.71, 28.45±2.82 vs 43.51±4.38, 12.16±2.13 vs 20.53±3.65, respectively), and significantly increased in the autophagy induction group (20.49±3.68 vs 9.01±1.24, 26.58±3.96 vs 11.83±1.71, 73.18±7.15 vs 43.51±4.38, 41.11±8.87 vs 20.53±3.65). Compared with the autophagy inhibition group and the ZNF580-silenced group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells of the autophagy inhibition+ZNF580 silencing group were significantly decreased (Col-Ⅰ: 3.36±1.25 vs 5.73±0.62 and 5.57±0.35; Col-Ⅲ: 4.15±0.16 vs 6.24±0.90 and 6.71±0.34; FN: 18.31±2.00 vs 26.46±1.18 and 27.09±2.01; TNF-α: 6.81±0.46 vs 9.96±1.87 and 10.62±0.65). All the above differences were statistically significant (all P value <0.05). Conclusions:The transcription factor ZNF580 could positively regulate the levels of autophagy and ECM secretion in PANC1 cells. The combined application of ZNF580 gene silencing and autophagy inhibitors can significantly inhibit ECM secretion in PANC1 cells.
2.Effects of zinc finger transcription factor 580 in regulating autophagy and extracelluar matrix secretion of PANC1 cells
Baiqing LIU ; Wei XU ; Wencheng ZHANG ; Shihai XIA
Chinese Journal of Pancreatology 2025;25(5):355-360
Objective:To investigate the regulatory effect of zinc finger transcription factor 580 (ZNF580) gene on autophagy and extracellular matrix (ECM) secretion in human pancreatic cancer cells PANC1.Methods:PANC1 cells were transfected with 500 ng/ml short hairpin RNA-ZNF580 (shRNA-ZNF580) and a ZNF580 expression vector with a green fluorescent protein reporter gene (GFP-ZNF580) using lentiviral transfection to establish the ZNF580-silenced group and ZNF580-overexpression group, respectively. PANC1 cells were treated with 10 mmol/L rapamycin (RA), a cell autophagy inducer, and the autophagy inhibitor LY294002 for 2 hours to construct the autophagy-induced group and autophagy-inhibited group, respectively. The autophagy inhibition+ZNF580 silencing group was established by transfecting PANC1 cells with 500 ng/ml sh-ZNF580 using lentiviral transfection while simultaneously adding 10 mmol/L LY294002. PANC1 cells cultured in conventional medium served as control group. The expression levels of ZNF580 protein and autophagy-related proteins ATG7 and LC3 in PANC1 cells from each group were detected by Western blot. The expression changes of ECM secretion-related markers type I collagen (Col-Ⅰ), Col-Ⅲ, fibronectin (FN), and tumor necrosis factor-α (TNF-α) in PANC1 cells were measured by ELISA.Results:Compared with control group, the protein expression levels of ATG7, LC3-Ⅰ, and LC3-Ⅱ in PANC1 cells of the ZNF580-silenced group were significantly decreased (0.40±0.04 vs 0.81±0.13, 0.66±0.08 vs 2.0±0.45, 0.78±0.10 vs 1.89±0.23), while they were significantly increased in the ZNF580-overexpression group (2.07±0.17 vs 0.83±0.09, 1.21±0.37 vs 0.88±0.09, 0.77±0.16 vs 0.37±0.06). The protein expression level of ZNF580 in PANC1 cells of the autophagy inhibition group was significantly down-regulated compared with the control group (0.40±0.15 vs 1.07±0.18), while it was significantly up-regulated in the autophagy induction group (1.59±0.25 vs 0.67±0.09). Compared with the control group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells were significantly decreased in the ZNF580-silenced group (5.02±0.81 vs 8.38±0.83, 6.17±0.83 vs 10.73±1.69, 28.66±2.47 vs 45.20±4.31, 10.09±1.32 vs 19.48±2.77), which were significantly increased in the ZNF580-overexpression group (19.28±2.05 vs 8.38±0.83, 28.29±5.96 vs 10.73±1.69, 103.22±6.37 vs 45.20±4.31, 46.78±6.96 vs 19.48±2.77), significantly decreased in the autophagy inhibition group (5.10±0.66 vs 9.01±1.24, 7.22±0.67 vs 11.83±1.71, 28.45±2.82 vs 43.51±4.38, 12.16±2.13 vs 20.53±3.65, respectively), and significantly increased in the autophagy induction group (20.49±3.68 vs 9.01±1.24, 26.58±3.96 vs 11.83±1.71, 73.18±7.15 vs 43.51±4.38, 41.11±8.87 vs 20.53±3.65). Compared with the autophagy inhibition group and the ZNF580-silenced group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells of the autophagy inhibition+ZNF580 silencing group were significantly decreased (Col-Ⅰ: 3.36±1.25 vs 5.73±0.62 and 5.57±0.35; Col-Ⅲ: 4.15±0.16 vs 6.24±0.90 and 6.71±0.34; FN: 18.31±2.00 vs 26.46±1.18 and 27.09±2.01; TNF-α: 6.81±0.46 vs 9.96±1.87 and 10.62±0.65). All the above differences were statistically significant (all P value <0.05). Conclusions:The transcription factor ZNF580 could positively regulate the levels of autophagy and ECM secretion in PANC1 cells. The combined application of ZNF580 gene silencing and autophagy inhibitors can significantly inhibit ECM secretion in PANC1 cells.
3.Phenotype and genotyping of Yersinia pestis in China
Xuefei ZHANG ; Jian HE ; Wenyuan XIN ; Juan JIN ; Xiaoyan YANG ; Sheng LI ; Qi ZHANG ; Youquan XIN ; Qingwen ZHANG ; Ruixia DAI ; Baiqing WEI
Chinese Journal of Endemiology 2023;42(7):517-524
Objective:To study the phenotype and genotype distribution of Yersinia pestis ( Y. pestis) in different natural foci of plague in China, so as to provide scientific basis for plague prevention and control. Methods:A total of 2 184 strains of Y. pestis isolated from different time periods, regions, hosts and vectors in 11 plague natural foci of China since 1943 were selected for biochemical type identification, glycolysis test, virulence factor test [capsule antigen (F1), pesticin Ⅰ (Pst Ⅰ), virulence antigen factor (VWa), pigmentation factor (Pgm)], different region (DFR) typing and clustered regularly interspaced short palindromic repeats (CRISPR) typing. Results:There were 16 biochemical types of Y. pestis in the natural foci of plague in China, and each biochemical type showed obvious regional distribution in each foci. Most strains were positive for ass hide glue glycolysis (89.79%, 1 961/2 184), maltose (80.13%, 1 750/2 184), glycerol (94.23%, 2 058/2 184), and denitrification (82.78%, 1 808/2 184), and negative for rhamnose (88.78%, 1 939/2 184) and melibiose (85.62%, 1 870/2 184). Virulence factor test results showed that 99.95% (2 183/2 184) of Y. pestis were F1 positive; 99.73% (2 178/2 184) of Y. pestis can produce Pst Ⅰ; 73.31% (1 601/2 184) of Y. pestis were VWa positive and 26.69% (583/2 184) were VWa negative; Pgm positive strains accounted for 72.62% (1 586/2 184), Pgm negative strains accounted for 21.52% (470/2 184), and Pgm mixed type strains accounted for 5.86% (128/2 184). According to DFR typing results, there were 52 genotypes in 2 184 strains of Y. pestis, of which 19 were major genotypes and 33 were minor genotypes. CRISPR typing revealed 16 major genotypes, of which 7 were newly discovered. Conclusion:The phenotypes and genotypes of Y. pestis in various natural foci of plague in China are diverse and have geographical distribution characteristics.
4.Studies on drug and disinfectant resistance related genes of Yersinia pestis in China
Ruixia DAI ; Jian HE ; Xiaoyan YANG ; Youquan XIN ; Sheng LI ; Juan JIN ; Qi ZHANG ; Jixiang BAI ; Baiqing WEI
Chinese Journal of Endemiology 2021;40(1):27-31
Objective:To understand whether there are drug resistant and disinfectant resistant Yersinia pestis strains in China, and to provide accurate information for clinical treatment of plague. Methods:A total of 2 753 Yersinia pestis strains isolated from 10 natural plague foci in China from 1943 to 2016 were collected. According to National Center for Biotechnology Information (NCBI) released sequences of aminoglycoside streptomycin resistant genes strA, strB, β-lactam antibiotics resistant genes TEM, SHV and CTX-M, sulfamilamide resistant genes sul1, sul2 and sul3, and disinfectant resistant gene qacE△1-sul1, a pair of primers of each gene was designed for above-mentioned genes. Genomic DNA of 2 753 strains of Yersinia pestis was extracted, and the 9 target genes of all DNA samples were amplified by PCR. Results:Negative and positive controls of PCR detection were established. No corresponding target bands of aminoglycoside streptomycin resistant genes strA, strB, β-lactam antibiotics resistant genes TEM, SHV and CTX-M, sulfamilamide resistant genes sul1, sul2 and sul3, and disinfectant resistant gene qacE△1-sul1 were found in the DNA samples of 2 753 strains of Yersinia pestis.Conclusion:The above-mentioned genes of drug resistance and disinfectant resistance have not been detected in Yersinia pestis of China, but the monitoring of drug resistance of Yersinia pestis still needs to be carried out continuously.
5.Etiology of Yersinia pestis in a plague natural foci of Qinghai-Tibet Plateau
Ruixia DAI ; Jian HE ; Youquan XIN ; Xiaoyan YANG ; Cunxiang LI ; Haoming XIONG ; Juan JIN ; Jianguo YANG ; Baiqing WEI ; Wei LI
Chinese Journal of Endemiology 2021;40(6):454-459
Objective:To analyze the pathogenic characteristics of Yersinia pestis in a plague natural foci in Qinghai-Tibet Plateau. Methods:In this study, 1 378 strains of Yersinia pestis isolated from different regions, hosts and vectors in Qinghai-Tibet Plateau from 1954 to 2016 were taken as the research objects. Phenotypic characteristics, plasmid spectrum and genotype of the strains were studied by using conventional techniques and molecular biological techniques. The etiology and geographical distribution of the plague were studied. Results:There were 6 biochemical types of Yersinia pestis in Qinghai-Tibet Plateau, namely Qinghai-Tibet Plateau, Qilian Mountain, Gangdis Mountain, Kunlun Mountain A, Kunlun Mountain B and Chuanqing Plateau. This study found that the Qinghai-Tibet Plateau type strain was not only distributed in north Tibet Plateau, but also distributed in south Tibet, and the distribution of Gangdis Mountain type strain extended to south Tibet. Four virulence factors (capsule antigen, yersinin, virulence antigen and pigmentation factor) were found in 79.97% (1 102/1 378) Yersinia pestis. The results also showed that there were 12 kinds of plasmids carried by Yersinia pestis strains in Qinghai-Tibet Plateau, which constituted 17 kinds of plasmid spectrum. There were 3 kinds of the largest plasmids with taxonomic properties, forming their respective relatively independent distribution areas. The study of different regions (DFR) type showed that 5, 8, 14, 19, 32 and 44 of 1 378 strains were the main genotypes, and the main genome types had obvious geographical distribution. Conclusions:All the tested strains have the characteristics of plague pathogen in Qinghai-Tibet Plateau. The polymorphism of the main hosts, vectors and the ecological landscape of plague geography in the plague foci in Qinghai-Tibet Plateau may lead to the diversity of biochemical characters, plasmid spectrum and geno types of Yersinia pestis.
6.Establishment and application of detection method for minimum inhibitory concentration of Yersinia pestis
Jian HE ; Xiaoyan YANG ; Youquan XIN ; Juan JIN ; Sheng LI ; Qi ZHANG ; Jixiang BAI ; Hailian WU ; Yong JIN ; Xuefei ZHANG ; Ruixia DAI ; Baiqing WEI
Chinese Journal of Endemiology 2020;39(11):842-846
Objective:To establish the minimum inhibitory concentration (MIC) detection method of Yersinia pestis by determining MIC of 11 kinds of antibiotics against Yersinia pestis, to master the inhibition range of common antibiotics on Yersinia pestis, and provide baseline data for clinical treatment of plague. Methods:According to Clinical Labor Standard Institution (CLSI), the agar plate dilution method was used to determine the MIC of 11 kinds of antibiotics against 118 strains of Yersinia pestis, including ofloxacin, ciprofloxacin, trimethoprim-sulfamethoxazole, kanamycin, streptomycin, ceftriaxone, ampicillin, chloramphenicol, spectinomycin, cefuroxime and tetracycline. MIC 50 and MIC 90 (the minimum concentration of drug which could inhibit 50% and 90% of bacterial growth) were calculated. The consistency was observed by comparing the results with those of the disk diffusion method. One hundred and eighteen strains of Yersinia pestis were isolated from natural plague foci of Qinghai Province and preserved by Qinghai Institute for Endemic Disease Prevention and Control. Results:Among 118 strains of Yersinia pestis tested, no single or multiple strains of Yersinia pestis resistant to 11 kinds of antibiotics were found, which was consistent with the results of the disk diffusion method. The MIC 50 and MIC 90 of 11 kinds of antibiotics against 118 strains of Yersinia pestis were obtained. Conclusions:The MIC detection method of Yersinia pestis is successfully established. This method can be used to measure the MIC of antibiotics against Yersinia pestis in high throughput and evaluate the sensitivity of Yersinia pestis to antibiotics. It is an efficient, economical and practical experimental method.
7. Study on the characteristics of major birth defects in 1.69 million cases of fetus in Guangxi Zhuang Autonomous Region
Baiqing DONG ; Biyan CHEN ; Qiuyu LIANG ; Sheng HE ; Wei LYU ; Botong LIU ; Yangjin ZUO ; Li LIN ; Hui WEI ; Jie WEI ; Xiuning HANG ; Xiaoxia QIU
Chinese Journal of Epidemiology 2019;40(12):1554-1559
Objective:
Tracking the information on 1.69 million fetal cases across Guangxi Zhuang Autonomous Region (Guangxi) so as to study the occurrences of total and major birth defects in order to evaluate the ability on related prevention and control programs in Guangxi.
Methods:
Using the self-developed "Gui Women’s System" to establish a database of 1.69 million fetal cases in Guangxi and to analyze the distribution of time, space and population, as well as the outcomes of pregnancy, using the big data.
Results:
During the 29 months of observation, the overall live birth rate was 99.25
8. Study on the distribution and related factors of birth weight of live births in Guangxi Zhuang Autonomous Region of China from 2016 to 2018
Baiqing DONG ; Jianpeng YOU ; Qiuyu LIANG ; Wei LYU ; Jinfeng MA ; Hongwei WEI ; Hai LI
Chinese Journal of Preventive Medicine 2019;53(12):1259-1264
Objective:
To investigate the distribution and related factors of birth weight of live births and full-term infants in Guangxi Zhuang Autonomous Region of China.
Methods:
Based on Guangxi women and children information system from 2016 to 2018, a large real-time database about maternal and live-birth information was established. It covered 1 712 midwifery institutions in Guangxi. A total of 2 394 240 cases of live births were collected and 2 243 129 cases of which were full-term infants. The multivariate logistic regression model was used to analyze the related factors of low birth weight.
Results:
The birth weight of 2 394 240 live births, (3 123.49±461.08) g, in Guangxi was approximately normal distribution with a peak distribution to the left. The incidence of low birth weight was 8.05%, and the incidence of macrosomia was 2.07%. The incidence of low birth weight was 10.92% for the puerpera with body mass index (BMI, kg/m2) <18.5, 16.82% for the puerpera with height
9.Characteristics of etiology and drug and disinfectant resistant genes of Yersinia pestis in natural plague foci of Haixi Prefecture, Qinghai Province
Sheng LI ; Xiaoyan YANG ; Jian HE ; Cunxiang LI ; Haoming XIONG ; Youquan XIN ; Juan JIN ; Qi ZHANG ; Jixiang BAI ; Baiqing WEI ; Ruixia DAI
Chinese Journal of Endemiology 2019;38(1):15-20
Objective To analyze the biological characteristics of Yersinia pestis strains in Haixi Prefecture,Qinghai Province,in order to provide a scientific basis for plague prevention and control in future.Methods Totally 181 strains were separated from variety kinds of host in Haixi Prefecture,Qinghai Province from 1957 to 2011,and these strains were conducted biochemical test,virulence factor evaluation,plasmid analysis,different region (DFR) genotyping,drug and disinfectant resistant genes detection;79 of the 181 strains were examined by toxicity test and classified according to the criteria (minimum lethal dose:MLD≤ 10 000 was velogenic strain,10 000 < MLD < 100 000 was moderate virulence strain,MLD ≥ 100 000 was hypovirulent strain).Results According to six biochemical typing about gelatin candy,rhamnose,maltose,melibiose,glycerin and denitrification,the 181 strains of Yersinia pestis were antique biovar and Qing-Tibet Plateau ecotype.Aproportion of 81.22% (147/181) of Yersiniapestis strains contained all the four virulence factors (F1,Pst Ⅰ,VW,Pgm).Totally 63.54% (115/181) of the strains contained 3 kinds of plasmid-6 × 106,45 × 106,and 52 × 106;31.49% (57/181) of the strains contained 3 kinds of plasmid-6 × 106,45 × 106,and 65 × 106.The strains had 8 genomovars,and were given priority to genomovar 8 (109 strains),secondly,genomovar 32 (33 strains),genomovar 5 (20 strains),genomovar 1b(i4 strains),genomovar 44 (2 strains),genomovar 7 (1 strain),genomovar 37 (1 strain),and genomovar 49 (1 strain).Among the 181 Yersinia pestis strains,strains with genes related to streptomycin resistance,sulfanilamide resistance,beta lactam resistance and disinfectant resistance were not found;and 75 of 79 strains were velogenic strains by toxicity test (MLD ≤ 10 000),accounted for 94.94% (75/79).Conclusion The strains separated in Haixi Prefecture,Qinghai Province have the characteristics of Qinghai-Tibet Plateau plague's pathogen and have strong toxicity;all strains don't have the characteristics of drug and disinfectant resistance genes.
10.Cyst of the hip ligamentum teres: a case report and literature review
Chunbao LI ; Baiqing ZHANG ; Yujie LIU ; Wei QI ; Bo HU ; Feng GAO ; Ming LU ; Yufeng LIU ; Mingyang AN
Chinese Journal of Orthopaedics 2019;39(7):436-439
According to the literature review, no cyst of the hip ligamentum teres has been reported. In this study, a 50?year?old female patient was admitted to the hospital because of sprained left hip pain with movement restriction for 15 months. The unilateral hip joint MRI showed cyst of the hip ligamentum teres, which was diagnosed as left hip joint cyst in the outpatient de?partment. Thus, the patient was admitted to the hospital. Physical examination showed that the left hip joint movement was slightly restricted with positive FADDIR test and FABER test. Arthroscopic surgical examination revealed a 1.5 cm×1.0 cm cyst at the lo?cation of the round ligament which was then cleared with a planer knife and radiofrequency. The treatment results were satisfacto?ry. The present study reviewed the literature about anatomical and biomechanical physicochemical properties of the ligamentum teres. The function of the ligamentum teres is important for the hip joint. In diagnosis of hip disease, unilateral hip MRI plays an important role in accurate diagnosis of such diseases. The hip arthroscopy provide a minimal invasive technique in treating cyst of the hip ligamentum teres. With the application of the hip arthroscopy, it will undoubtedly improve the diagnosis and treatment of hip diseases.

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