1.TBX1 inhibits the activation of MAPK and PI3K signaling pathway and reduces the proliferation of colorectal cancer cells thorough activating PARK2
Jingni ZHANG ; Tong HUANG ; Mengge BAI ; Jianling WANG ; Man ZHAO
Cancer Research and Clinic 2024;36(11):808-816
Objective:To investigate the effect of the transcription factor TBX1 on the proliferation of colorectal cancer cells and to explore potential molecular mechanisms.Methods:The mRNA and protein levels of TBX1 in colorectal cancer cell lines HCT116, RKO, SW480, HT29, and LOVO were detected by using reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Colorectal cancer cell lines HCT116 and SW480 cells with low TBX1 expression were transfected with either a pcDNA3.1 plasmid containing TBX1 mimics (TBX1 overexpression group) or an empty pcDNA3.1 plasmid (the control group). LOVO cells with high TBX1 expression were transfected with small interfering RNA (siRNA) targeting TBX1 including si-TBX1-8604A, si-TBX1-8604B, and a negative control siRNA (si-NC), which were treated as si-TBX1-8604A group, si-TBX1-8604B group, and si-NC group. qRT-PCR was used to detect the expressions of transcriptional level TBX1 and PARK2, and Western blot was used to detect the protein levels of TBX1, PARK2, and key factors in the MAPK and PI3K signaling pathways. Methyl Thiazolyl Tetrazolium (MTT) assay and cell colony formation assay were used to detect the cell proliferation. Combining literatures and the JASPAR database, 2 binding sites of TBX1 in the PARK2 promoter region were predicted. Chromatin immunoprecipitation assay was employed to verify the binding sites of TBX1 to PARK2 in HCT116 and SW480 cells. Dual luciferase reporter gene assay was used to verify the targeting relationship between TBX1 and PARK2. The expression of TBX1 and PARK2 in colon cancer tissues was analyzed by using the Cancer Genome Atlas (TCGA) database (September 2023).Results:High TBX1 expression in HCT116 and SW480 cells transfected with TBX1 mimics plasmid was confirmed by qRT-PCR and Western blot, while TBX1 expression was successfully knocked down in LOVO cells transfected with siRNA targeting TBX1. MTT assay indicated that the absorbance values for HCT116 cells in TBX1 overexpression group on d1, d3, d5, and d7 after inoculation, and for SW480 cells on d3, d5, and d7 after inoculation were lower than those in the control group, and the differences were statistically significant (all P < 0.01). LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group exhibited higher absorbance values than the si-NC group on d1, d3, d5, and d7 after inoculation, and the differences were statistically significant (all P < 0.05). Cell colony formation assay revealed that after 14 d, the colony number of HCT116 cells [(387±9) vs. (843±13)] and SW480 cells [(413±9) vs. (931±15)] in TBX1 overexpression group was lower than that in the control group, and the differences were statistically significant (all P < 0.05). The colony number of LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group was (493±77) and (470±32), respectively, which was higher than that in the si-NC group (349±26), and the differences were statistically significant (all P < 0.05). The protein relative expression levels of p-ERK and p-AKT S473 in HCT116 and SW480 cells in TBX1 overexpression group were lower than those in the control group, while protein relative expression levels of p-ERK and p-AKT S473 in LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group were higher than those in the si-NC group, and the differences were statistically significant (all P < 0.05). The relative expression level of PARK2 mRNA in HCT116 and SW480 cells (all P < 0.01) and the protein level in the overexpression group were higher than those in the control group. Chromatin immunoprecipitation assay demonstrated that the enrichment times of TBX1 binding to 2 sites of PARK2 intron in HCT116 and SW480 cells in TBX1 overexpression group were higher than that in the control group, and the differences were statistically significant (all P < 0.05). Dual luciferase reporter gene assay showed that the relative luciferase activity of HCT116 and SW480 cells co-transfected with pcDNA3.1 plasmid containing TBX1 mimics and pGL3 plasmid containing PARK2 mimics was higher than that of cells co-transfected with empty pcDNA3.1 and pGL3 plasmids, co-transfected with empty pcDNA3.1 plasmid and pGL3 plasmid containing PARK2 mimics, co-transfected with pcDNA3.1 plasmid containing TBX1 mimics and empty pGL3 plasmid, and the differences were statistically significant (all P < 0.05). Spearman analysis showed that there was a positive correlation between transcriptional level TBX1 and PARK2 in colon cancer tissues (288 cases) in TCGA database ( r = 0.226, P < 0.001); and the relative expression level of PARK2 mRNA in colon cancer tissues (383 cases) was lower than that in normal intestinal tissues (50 cases), and the difference was statistically significant ( P < 0.001). Conclusions:Elevated expression of transcriptional factor TBX1 inhibits the proliferation of colorectal cancer cells, potentially by activating the downstream target gene PARK2 and inhibiting the phosphorylation of ERK and AKT in the MAPK and PI3K signaling pathways, ultimately affecting the activation of these pathways.
2.A primary application of 99mTc-MDP whole body bone scintigraphy in evaluation of bone and joint injuries in patients with early brucellosis
Hongwei YIN ; Qiang LI ; Liqing XU ; Qiang GUO ; Hongmei XUE ; Jianling WANG ; Jiquan LI ; Zhijun ZHAO
Chinese Journal of Endemiology 2023;42(2):148-151
Objective:To investigate the diagnostic value of 99mTc-methylenediphosphonate(MDP) whole body bone scintigraphy in early brucellosis patients with bone and joint injuries. Methods:According to the Diagnosis for Brucellosis (WS 269-2019), combined with epidemiological history, clinical manifestations and serological tests, from November 2020 to April 2021, 15 early brucellosis patients (the course of disease was within 6 months) who had not received any drug treatment diagnosed at the Department for Brucellosis Prevention and Control, Qinghai Institute for Endemic Disease Prevention and Control were selected as the research subjects, and 99mTc-MDP whole body bone scintigraphy was performed on the patients to evaluate the images and analyze the pathological changes. Results:The 99mTc-MDP whole body bone scintigraphy of 15 patients with early brucellosis showed abnormalities, and the abnormal concentration of radionuclides mainly occurred in the 8th to 12th thoracic vertebrae (T8-12), the 1st to 2nd lumbar vertebrae (L1-2) and L4-5. Among them, the thoracic vertebrae abnormalities were T8, T9, T10, T11 and T12 in 1 case each; lumbar vertebrae abnormalities were 1 case of L1, 1 case of L2, 4 cases of L4, and 5 cases of L5. Conclusions:The 99mTc-MDP whole body bone scintigraphy is abnormal in patients with early brucellosis. Bone scintigraphy has certain value in the diagnosis of bone and joint injuries in patients with early brucellosis.
3.Genotyping and traceability analysis of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau by MLVA
Li MA ; Xuefei ZHANG ; Hongmei XUE ; Aiping ZHANG ; Lingling REN ; Teng QI ; Yuanbo ZHAO ; Jianling WANG ; Xuxin YANG ; Jiquan LI
Chinese Journal of Endemiology 2023;42(4):269-273
Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.
4.HBV infection among blood donors from 18 domestic blood stations of prefecture-level cities
Dingding WANG ; Youhua SHEN ; Jianling ZHONG ; Hui ZHANG ; Zhibin TIAN ; Lin BAO ; Huixia ZHAO ; Jian ZHANG ; Peng WANG ; Yanqin HE ; Wei ZHANG ; Li LI ; Hao LI ; Dexu CHU ; Ying WANG ; Xin ZHANG ; Shouguang XU ; Min HUANG ; Yan QIU
Chinese Journal of Blood Transfusion 2023;36(2):172-176
【Objective】 To analyze the hepatitis B virus (HBV) infection data of blood donors from 18 domestic blood stations, so as to investigate the HBV infection situation of blood donors. 【Methods】 The positive rate of HBV and its distribution characteristics of regions, the percentage of HBsAg+ ELISA in first-time vs repeated blood donors, and the percentage of HBsAg-/HBV DNA+ blood donors of 18 domestic blood stations during 2017 to 2020 were collected from the Working Platform for Practice Comparison of Blood Centers, and the HBV infection among blood donors were statistically analyzed. 【Results】 From 2017 to 2020, the positive rate of HBV in blood donors among 18 domestic blood stations was 13.48/10 000-144.02/10 000, with the average HBV positive rate in eastern, central and western region at 26.14/10 000, 51.98/10 000 and 41.00/10 000, respectively. The HBsAg+ rate by ELISA among first-time and repeated blood donors was 14.55/10 000-305.39/10 000 vs 1.04/10 000-87.43/10 000 The HBsAg-/HBV DNA+ yield was 1.80/10 000-35.31/10 000. 【Conclusion】 The distribution of HBV infection in blood donors has regional characteristics, and HBV prevalence was low in repeated blood donors. HBsAg ELISA combined with HBV DNA detection can better ensure blood safety.
5.Metabolic Disease Management Guideline for National Metabolic Management Center(2nd edition)
Weiqing WANG ; Yufan WANG ; Guixia WANG ; Guang NING ; Dalong ZHU ; Ping LIU ; Libin LIU ; Jianmin LIU ; Zhaoli YAN ; Xulei TANG ; Bangqun JI ; Sunjie YAN ; Heng SU ; Jianling DU ; Sheli LI ; Li LI ; Shengli WU ; Jinsong KUANG ; Yubo SHA ; Ping ZHANG ; Yifei ZHANG ; Lei CHEN ; Zunhai ZHOU ; Chao ZHENG ; Qidong ZHENG ; Zhongyan SHAN ; Dong ZHAO ; Zhigang ZHAO ; Ling HU ; Tingyu KE ; Yu SHI ; Yingfen QIN ; Mingjun GU ; Xuejiang GU ; Fengmei XU ; Zuhua GAO ; Qijuan DONG ; Yi SHU ; Yuancheng DAI
Chinese Journal of Endocrinology and Metabolism 2023;39(6):538-554
The latest epidemiological data suggests that the situation of adult diabetes in China is severe, and metabolic diseases have become significant chronic illnesses that have a serious impact on public health and social development. After more than six years of practice, the National Metabolic Management Center(MMC) has developed distinctive approaches to manage metabolic patients and has achieved a series of positive outcomes, continuously advancing the standardized diagnosis and treatment model. In order to further improve the efficiency, based on the first edition, the second edition guideline was composed by incorporating experience of the past six years in conjunction with the latest international and domestic guidelines.
6.Analysis of surveillance results of human brucellosis in Qinghai Province in 2019 and 2020
Zhijun ZHAO ; Qiang LI ; Jiquan LI ; Jianling WANG ; Li MA ; Hongmei XUE ; Yu SHI ; Di FAN ; Yumin QIN ; Yuanbo ZHAO ; Guang TIAN ; Liqing XU
Chinese Journal of Endemiology 2022;41(3):200-203
Objective:To master the epidemic trend of human brucellosis in Qinghai Province, so as to provide basis for scientific prevention and control of the disease.Methods:In 2019 and 2020, at the national and provincial brucellosis monitoring sites in Qinghai Province, a total of 18 counties (cities and districts, hereinafter referred to as counties), no less than 400 serum samples were sampled every year for brucellosis Rose-Bengal plate agglutination test (RBPT) and serum tube agglutination test (SAT), which would be tested and judged according to the criteria of "Diagnosis for Brucellosis" (WS 269-2019).Results:In 2019, a total of 1 612 people were monitored in national brucellosis monitoring sites, 93 were RBPT positive, 54 were SAT positive, 54 were diagnosed, and the prevalence rate was 3.35% (54/1 612). In 2020, 1 677 people were monitored in national brucellosis monitoring sites, 151 were RBPT positive, 80 were SAT positive, 80 were diagnosed, and the prevalence rate was 4.77% (80/1 677). There were significant differences in RBPT positive rate, SAT positive rate and prevalence rate among national monitoring sites between the two years (χ 2 = 12.52, 4.24, 4.24, P < 0.05). In 2019, a total of 6 043 people were monitored in provincial brucellosis monitoring sites, 128 were RBPT positive, 91 were SAT positive, 87 were diagnosed, and the prevalence rate was 1.44% (87/6 043). In 2020, 5 664 people were monitored, 108 were RBPT positive, 59 were SAT positive, 52 were diagnosed, and the prevalence rate was 0.92% (52/5 664). There was no significant difference in RBPT positive rate among provincial monitoring sites between the two years (χ 2 = 0.66, P = 0.416), and the differences in SAT positive rate and prevalence rate were statistically significant among provincial monitoring sites between the two years (χ 2 = 4.98, 14.57, P < 0.05). Conclusion:In 2019 and 2020, there are human brucellosis in national and provincial brucellosis monitoring sites in Qinghai Province.
7.Analysis of serum erythropoietin test results of adult patients with Kaschin-Beck disease in Qinghai Province
Yu SHI ; Qiang LI ; Xin ZHOU ; Hongmei XUE ; Jianling WANG ; Guanglan PU ; Cuiling LA ; Lansheng HU ; Liqing XU ; Jiquan LI ; Zhijun ZHAO ; Lihua WANG
Chinese Journal of Endemiology 2022;41(6):437-439
Objective:To analyze the results of serum erythropoietin (EPO) in adults patients with Kaschin-Beck disease (KBD) in Qinghai Province.Methods:According to the "Diagnosis of Kaschin-Beck Disease" (WS/T 207-2010), by using clinical examination and X-ray, adults over 20 years old in KBD areas of Xinghai County and Guide County, Hainan Tibetan Autonomous Prefecture, Qinghai Province, were divided into KBD case group ( n = 109) and internal control group ( n = 95) in July 2019. At the same time, healthy people were selected as external control group ( n = 90) in Xunhua County. Then 2 ml fasting cubital venous blood was collected from the target population to separate serum. The serum EPO level was determined by enzyme-linked immunosorbent assay (ELISA). Results:There was no significant difference in age and sex ratio among the 3 groups ( F = 0.73, P = 0.484; χ 2 = 1.03, P = 0.611). There was significant difference in serum EPO levels among the 3 groups [KBD case, internal and external control groups: (30.74 ± 26.23), (19.73 ± 11.53) and (10.83 ± 4.48) U/L, F = 26.51, P < 0.001]. Multiple comparisons showed that there were statistically significant differences in serum EPO levels between KBD case group and the internal and external control groups ( P < 0.05), but there was no significant difference between the internal and external control groups ( P > 0.05). Conclusions:The serum EPO level in adult KBD patients in Qinghai Province is increased significantly.
8.Classification of lower leg osteofacial compartment syndrome
Ying LI ; Junsheng YANG ; Zhiwei YANG ; Liangcheng TONG ; Lei ZHAO ; Jianling WANG ; Zhongyang SUN ; Qing XUE
Chinese Journal of Trauma 2022;38(5):458-461
Osteofacial compartment syndrome (OCS) is one of the serious complications in traumatic orthopedics. If not treated in time, OCS may result in irreversible damage to nerve and muscle,even amputation or death in serious condition. 5P presents to be the classic clinical diagnosis of OCS, but it is highly subjective and cannot timely and accurately judge the progression of the disease. Intracompartment pressure manometry is the main auxiliary method for the diagnosis of OCS. Although there are many manometry methods, there is still no authoritative pressure threshold as the diagnosis standard. Clinicians often aggressively perform fasciotomy to avoid serious complications, leading to unnecessary fasciotomy. The authors retrospectively reviewed the data of patients with OCS treated at Air Force Hospital of Eastern Theater of PLA from March 2010 to March 2020 and found that some patients with OCS had gradual alleviation of clinical symptoms after appropriate conservative treatments such as brace releasing, limb stabilization and swelling subsidence, with no need of fasciotomy. However, the symptoms of some patients progressively aggravated after the above-mentioned traditional treatments and timely fasciotomy was required. The authors graded the severity of OCS and proposed for the first time the OCS grading criteria according to quantitative clinical results and quantitative indicators such as ratio of mean blood flow velocity of bilateral arteries and pulse wave changes, aiming to take corresponding intervention measures for patients with different OCS classifications, carry out more precise treatment and avoid unnecessary fasciotomy.
9.Annual financial expenditure in 24 domestic blood stations: a comparative analysis
Huixia ZHAO ; Pengkun WANG ; Hongjun CAI ; Lina HE ; Qizhong LIU ; Feng YAN ; Jianhua LI ; Jiankun MA ; Jianling ZHONG ; Chaochao LV ; Yu JIANG ; Qingpei LIU ; Li LI ; Jian ZHANG ; Weitao YANG ; Wei ZHANG ; Zhenxing WANG ; Peng WANG ; Wenjie HUANG ; Qingjie MA ; Youhua SHEN ; Zhibin TIAN ; Meihua LUN ; Mei YU
Chinese Journal of Blood Transfusion 2022;35(9):947-949
【Objective】 To study the annual financial expenditure in blood stations with different scales, and to establish the regression equation between blood collection units and total expenditure. 【Methods】 The annual total expenditure, the per capita cost of serving population, as well as the collection units of whole blood and apheresis platelet of 24 blood stations were collected. The financial expenditure required for collecting 10 000U blood was calculated.The statistical analysis was carried out with SPSS statistical software. 【Results】 From 2017 to 2020, the total annual financial expenditure of 24 blood stations showed an upward trend. The total expenditure among blood stations was different. The per capita cost of servicing population in the areas where the 24 blood stations were located had been increasing year by year. The 24 blood stations were divided into two grades according to the blood collection volume as 50 000 U, and the relationship equation between the blood collection volume and the annual total expenditure had been established. After testing, each equation was effective(P<0.05); There was no difference in the financial expenditure required for collecting 10 000U blood among blood stations with different scales. 【Conclusion】 From 2017 to 2020, the blood stations with an annual collection volume more than 50 000 U demonstrated a higher financial expenditure and the per capita cost of serving population than those <50 000 U. The blood collection volume of blood stations is significantly correlated with the annual total expenditure and the per capita cost of serving population.
10.Protective effect of pre-perfusion with improved St.Thomas solution on skeletal muscle ischemia-reperfusion injury in dogs
Lei ZHAO ; Zhiwei YANG ; Chao XIA ; Guojun WANG ; Liangcheng TONG ; Junsheng YANG ; Jianling WANG ; Ying LI
Chinese Journal of Microsurgery 2022;45(6):656-663
Objective:To explore the protective effect and mechanism of improved St. Thomas solution on canine skeletal muscle ischemia-reperfusion injury (IRI).Methods:Between March 2021 and September 2021, in the experimental operating room at the Air Force Hospital of the PLA Eastern Theater Command, 16 Beagles were randomly divided into control group, IRI group, IRI+NS group, and improved St. Thomas group, 4 in each group. The canine skeletal muscle IRI model was established, and the canine vital signs were monitored by pre-perfusion with improved St. Thomas perfusate [potassium chloride (KCl), magnesium sulfate (MgSO 4), and NaHCO 3 (pH adjusted)]. The pathological damage of canine skeletal muscle was explored by hematoxylin eosin (HE) staining, electron microscope detection and tissue wet/dry weight ratio, and blood vessel density. Hypoxia performances were detected by labeling blood vessels and hypoxia-inducible factor-1α (HIF-1α). The IRI model of L6 rat myoblasts was established, and the components of St. Thomas perfusion solution were pre incubated to explore the effect on the inhibition of cell proliferation. And by detecting reduced nicotinamide adenine dinucleotide phosphate (NADPH), F2 isoprostane (F2-isoprostane), interleukin 1β(IL-1β), tumour necrosis factor alpha (TNF-α), myeloperoxide enzyme (MPO), glutathione peroxidase (GSH-Px), etc. to explore its protective mechanism. Statistical software SPSS 23.0 was used for statistical analysis, A P<0.05 was set as statistically significant. Results:In the improved St. Thomas group, the vital signs of the dogs were relatively stable, the amount of maintained dopamine was less, the histopathological structure of the gastrocnemius muscle tended to be intact, the swelling of tissue cells and mitochondria was significantly relieved, and the tissue wet/dry weight ratio was less than that in the IRI group ( P=0.046). Pre-incubated with therapeutic doses of MgSO 4 or NaHCO 3, the proliferation rate of L6 cells was higher than that of IRI group ( P<0.01, P=0.005), NADPH ( P=0.004, P=0.001), F2-isoprostane ( P<0.01, P=0.01), IL-1β ( P=0.02, P=0.015), TNF-α ( P<0.01, P<0.01), MPO ( P<0.01, P<0.01) were all lower than those in the IRI group, except GSH-Px that was higher than what in the IRI group ( P<0.01). Conclusion:Pre-perfusion of the improved St. Thomas solution can stabilise the vital signs of dogs in a short period of time. The solution can improve the state of skeletal muscle cells, improve tissue hypoxia, and reduce the damage of skeletal muscle tissue cells through anti-inflammatory and anti-oxidative stress.

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