1.Effect of Shenqi Yiliu Prescription Combined with Cisplatin on Tumor in Hepatoma H22-bearing Mice Based on PTEN/PI3K/Akt Signaling Pathway
Xin FENG ; Yongqiang DUAN ; Min BAI ; Yuping YANG ; Liren CAO ; Junrui HU ; Yanhua SI ; Jing CHEN ; Zihan GONG ; Lan MA
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(3):96-103
ObjectiveTo investigate the tumor-suppressing effect of Shenqi Yiliu prescription combined with cisplatin in hepatoma H22-bearing mice based on the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) pathway. MethodH22-bearing mice were prepared and randomized into model group, cisplatin group, and cisplatin combined with high-, medium-, and low-dose Shenqi Yiliu prescription groups, with 10 mice in each group. Another 10 healthy mice were randomly selected as normal group. Shenqi Yiliu prescription was given by gavage with the high, medium, low dose of 54.06, 27.03, 13.515 g·kg-1·d-1, respectively, and cisplatin (2.5 mg·kg-1) was administered by intraperitoneal injection, twice a week. Normal group and model group received normal saline. After 13 days of treatment, mice were killed and the tumor inhibition rate was calculated. The pathomorphological changes of tumor were observed based on hematoxylin-eosin (HE) staining, and enzyme-linked immunosorbent assay (ELISA) and immunofluorescence method were used to detect the content of cyclin-dependent kinase inhibitor 1A (p21) and cyclin-dependent kinase inhibitor 1B (p27) in tumor tissue of mice. The levels of PTEN, PI3K and phosphorylated protein kinase B (p-Akt) in tumor tissue were measured by Western blot. ResultCompared with the model group, cisplatin alone and cisplatin in combination with the high-, medium-, and low-dose Shenqi Yiliu prescription decreased tumor mass (P<0.05), particularly the cisplatin in combination with the high-dose Shenqi Yiliu prescription. Necrosis of the tumor tissue was observed in each group, especially the cisplatin combined with high-dose Shenqi Yiliu prescription group. As compared with the model group, cisplatin alone and cisplatin in combination with the high-, medium-, and low-dose Shenqi Yiliu prescription raised the expression of p21, p27, and PTEN (P<0.05) and lowered the expression of PI3K and p-Akt (P<0.05), particularly the cisplatin in combination with high-dose Shenqi Yiliu prescription. ConclusionShenqi Yiliu prescription may regulate the expression of key molecules in PTEN/PI3K/Akt signaling pathway, thereby upregulating the expression of downstream proliferation inhibitors p21 and p27, further suppressing the tumor in H22-bearing mice, and enhancing the effect of chemotherapy.
2.Establishment and identification of a mitochondrial tracking system
Lin LYU ; Sihan WANG ; Quan ZENG ; Han DUAN ; Zhuang MAO ; Changyao WANG ; Xuetao PEI ; Hua WANG ; Yanhua LI
Chinese Journal of Pharmacology and Toxicology 2023;37(12):928-935
OBJECTIVE To observe whether mitochondria can be transferred from mesenchymal stem cells(MSCs)to irradiated cells by establishing a mitochondrial fluorescent reporting system.METHODS The lentiviral vector pSIN-EF1α-COX8A-DsRed2(named COX8A-DsRed2)that might guide the expres-sion of red fluorescence protein in the membrane of mitochondria was constructed.A lentivirus(named Lv-COX8A-DsRed2)was prepared in 293T cell line.Dental pulp stem cells(DPSCs)(named DPSC-COX8A-DsRed2)was infected with Lv-COX8A-DsRed2.The intracellular expression of the red fluores-cence protein in DPSC was observed under fluorescence microcopy.The mitochondrial localization of the expressed red fluorescent probe in DPSC-COX8A-DsRed2 was confirmed according to TOMM20 immunostaining and MitoTracker Green staining results,which could specifically label mitochondria.The IEC-6 cells that received 10 Gy X-ray radiation were used as an injured cell model.The co-culture system was established by supplementing DPSC-COX8A-DsRed2 into the culture plate with the irradi-ated IEC-6 labelled by CFSE for 24 h.RESULTS The imaging results of fluorescent microcopy obser-vation showed that DPSC-COX8A-DsRed2 expressed the mitochondrial fluorescent reporting system,which was co-located with TOMM20 protein and Mito Tracker Green.The imaging results of confocal fluorescence microcopy showed that the mitochondria with red fluorescent protein were transferred from DPSC-COX8A-DsRed2 to the irradiated IEC-6 cells,suggesting that the established mitochondrial fluorescent reporting system could indicate mitochondrial transfer from donor cells to injured ones.CONCLUSION DPSC-COX8A-DsRed2 stably expressing the mitochondrial fluorescent reporting system is established,which can be used to track mitochondrial transfer.
3.Effect of electroacupuncture at Baihui ameliorated neurologic deficit and hemodynamic stability in rat model of post-cardiac arrest syndrome
Ruifeng ZENG ; Yuejia ZHONG ; Yanhua WU ; Peng WANG ; Chengzhi LAI ; Xiangqi LIU ; Yunbiao DUAN ; Zunjiang LI ; Tao YU ; Banghan DING
Chinese Critical Care Medicine 2022;34(12):1285-1290
Objective:To observe the results of electroacupuncture (EA) on the resuscitation of a rat model of asphyxia cardiac arrest (CA). And to explore its effect on the neurologic deficits and hemodynamic instability of post-cardiac arrest syndrome (PCAS).Methods:A total of 107 male SD rats were randomly divided into sham, CA, and EA groups. Each group received arterial catheterization and tracheal intubation. The sham group was not induced asphyxia. Asphyxial cardiac arrest was established by endotracheal tube clamping. Rats in the CA group received basic respiratory support and fluid resuscitation in return of spontaneous circulation (ROSC) and rats in the EA group received EA at Baihui based on the treatment of CA group after ROSC, with a dense-dispersed wave at frequencies of 4-20 Hz, while the current intensity was adjusted minimum to induce a twitch of the scalp, the course of treatment was 30 minutes. The baseline data, hemodynamics after ROSC, neurological deficit score (NDS), pathological changes of brain tissue, and levels of serum biomarker were recorded and compared among the three groups. The 72-hour survival of rats was analyzed by Kaplan-Meier survival curve. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of necrotic neurons in the hippocampal CA1 region of rat brain. Meanwhile, Nissl staining and TdT-mediated dUTP nick-end labeling (TUNEL) were used to detect cell apoptosis and injury.Results:Compared with the CA group, the mean arterial pressure (MAP) in the EA group increased significantly at 15 minutes after ROSC [mmHg (1 mmHg≈0.133 kPa): 125.00 (94.00, 136.25) vs. 92.00 (72.00, 122.50), P < 0.05]. There was no significant difference in the NDS score between the EA group and the sham group. Still, the NDS score of the rats in the CA group at 6 hours after ROSC were significantly lower than that in the sham group (46.00±10.61 vs. 80.00±0.00, P < 0.05). Kaplan-Meier survival curve analysis showed that EA did not improve the 72-hour survival rate of rats (100% in the sham group, 25% in the CA group, and 30% in the EA group, P > 0.05). The analysis by TUNEL showed that the apoptosis rate of neurons in CA1 region of the hippocampus in EA group at 6 hours after ROSC was significantly lower than that in CA group [(62.84±2.67)% vs. (71.29±3.70)%, P < 0.05]. Compared with the CA group, the level of serum S100 calcium binding protein B (S100B) in the EA group at 6 hours after ROSC was significantly lower (ng/L: 19.30±13.87 vs. 132.28±31.67, P < 0.05), but there were no significant differences in the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) between these two groups. Conclusions:In the present study, EA at Baihui can stabilize the hemodynamic, moreover, it has a particular neuroprotective effect on PCAS rats. Still, EA at Baihui does not reduce the systemic inflammatory response and improve the survival rate of rats, and its mechanism remains to be verified in further research.
4.Evaluation on the feasibility of total-body PET/CT imaging with short acquisition time in lungs and parenchymal organs
Minjie ZHAO ; Keyu ZAN ; Zhaoping CHENG ; Xiao CUI ; Leiying CHAI ; Kun LI ; Min GE ; Yanhua DUAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(12):713-718
Objective:To investigate the effects of total-body PET/CT imaging with short acquisition time on image quality and lesion detectability in lungs and parenchymal organs.Methods:Sixty patients (31 males, 29 females, age (61.1±11.8) years) with pulmonary nodules (PN) and 53 patients (29 males, 24 females, age (56.7±17.2) years) with parenchymal organ lesions (POL) who underwent total-body PET/CT imaging in the First Affiliated Hospital of Shandong First Medical University between October 2021 and April 2022 were retrospectively analyzed. The acquisition time with PET was 600 s, and the reconstructed images were divided into 6 groups based on different duration (30, 60, 120, 180, 300 and 600 s), namely G30, G60, G120, G180, G300 and G600 groups. The subjective analysis was carried out with the 5-point Likert scale in 3 aspects: the overall impression of image quality, noise, and lesion conspicuity. The objective analysis indicators included the SUV mean of the mediastinal blood pool (MBP); the SUV mean, standard deviation (SD) and signal-to-noise ratio (SNR) of the liver; SUV max and target-to-background ratio (TBR) of the lesions. Differences of the indicators among 6 groups were analyzed by Friedman test with Bonferroni correction. G600 served as the reference for the other 5 groups to test their lesion detectability. Results:The subjective image quality of different groups for PN and that of G120, G180, G300 groups for POL could meet the needs of clinical diagnosis in terms of the overall image quality, noise, and lesion conspicuity (all scores>3). There was no significant difference in the SUV mean of MBP among different time groups (median for PN: 1.52-1.56, median for POL: 1.35-1.47; χ2 values: 10.23, 11.02, both P>0.05). Difference was not found in SUV mean of the liver either (median for PN: 2.51-2.56, median for POL: 2.33-2.40; χ2 values: 8.35, 8.93, both P>0.05). The liver SD significantly increased along with the shortened acquisition time ( χ2 values: 400.99, 400.00, both P<0.001; z values: from -16.90 to -3.15, all P<0.003). The SNR significantly decreased along with the shortened acquisition time ( χ2 values: 397.32, 400.00, both P<0.001; z values: 2.98-16.90, all P<0.003). The SUV max (median for PN: 3.55-4.01, median for POL: 5.77-6.08; χ2 values: 8.58, 3.02, both P>0.05) and TBR (median for PN: 2.42-2.81, median for POL: 2.36-2.45; χ2 values: 9.83, 3.69, both P>0.05) of lesion were not significantly different among 6 groups. Taking G600 group as a reference, the lesion detection rates were 100% in G30 group and other 4 groups for PN (81/81) and in G120, G180, G300 groups for POL (80/80). Conclusion:Total-body PET/CT imaging with acquisition time of 30 s for lungs and that with acquisition time of 120 s for parenchymal organs are feasible for clinical use, with the PET image quality and lesion detectability maintained.
5.Influenza B virus monitoring and analysis of variation and evolution of hemagglutinin (HA) and neuraminidase (NA) genes in Jining city from 2017 to 2020
Tihui WANG ; Yanhua DUAN ; Shuqi HAN ; Shengnan WANG ; Yajuan JIANG ; Boyan JIAO
Chinese Journal of Microbiology and Immunology 2021;41(4):301-305
Objective:To analyze the variation and evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of influenza B virus circulating in Jining from 2017 to 2020.Methods:Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals and influenza outbreaks in Jining from 2017 to 2020 and tested for influenza B virus nucleic acid. After virus isolation, 20 representative strains of influenza B virus were selected to sequence the full length of HA and NA genes. Phylogenetic trees were constructed and the molecular characteristics were analyzed using bioinformatics software. Results:A total of 4 575 specimens were collected and 842 of them were positive for influenza virus, including 398 (8.7%, 398/4 575) influenza B virus-positive specimens. The positive rate of influenza B virus was 47.27% (398/842). The isolated influenza B virus strains of Victoria (BV) and Yamagata (BY) lineages from 2017 to 2020 shared 98.7%-98.8% and 98.5%-99.1% homology in HA gene with vaccine strains, respectively. The BV lineage strains isolated from 2018 to 2020 belonged to Victoria clade 1A branch and the BY lineage strains isolated from 2017 to 2018 belonged to Yamagata clade 3 branch. Mutations were detected in several antigenic sites, but not in the sites related to NA inhibitor resistance. Conclusions:Mutations in several antigenic sites caused antigenic changes in influenza B virus of BV and BY lineages, which might be related to the outbreaks of influenza B virus infection in Jining during 2017 to 2020.
6.Dose-effect relationship between serum polycyclic aromatic hydrocarbon adducts and serum complements among children in a city of East China
Zhenjie WANG ; Yang LIN ; Jingjing YIN ; Liya ZHANG ; Ting WANG ; Yanhua WANG ; Nan WU ; Fanling KONG ; Huawei DUAN
Chinese Journal of Preventive Medicine 2021;55(5):653-659
Objective:To investigate the dose-response relationship between serum polycyclic aromatic hydrocarbon adducts and serum complement C3 and C4 levels among children from a city in East China.Methods:In September 2016, two boarding schools in the air pollution exposure area and the control area (beyond the upwind of 30 km in the air pollution exposure area) in a city in East China were selected as the research site, and the eligible school-age children were recruited as the research objects. A total of 273 children were included, including 163 in the exposure group and 110 in the control group. The annual air pollutant data (PM 2.5, PM 10 and NO 2) of the two regions during the study period were collected. The exposure level of tobacco was evaluated by cotinine in urine. The levels of serum complement C3 and C4 were determined by automatic biochemical analyzer. The serum anti-7, 8, -dihydrodiol-9, 10-epoxide benzo[a]pyrene (BPDE)-albumin adduct levels were detected by ELISA. Linear regression model was used to explore the dose-response relationship between BPDE-albumin adducts and serum complement C3 and C4. Results:The age of 273 subjects was (13.67±0.37) years old, including 165 boys (60.4%). The average annual exposure levels of PM 2.5, PM 10 and NO 2 and the level of serum BPDE-albumin adducts in the exposure group were higher than those in the control group ( P<0.05). The results of linear regression model analysis showed that after adjusting age, sex, BMI z-score and urinary cotinine level, when the serum BPDE-albumin adduct level increased by 10%, the serum complement C4 level decreased by 1.2% ( P=0.017). After adjusting age, BMI z-score and urinary cotinine level, for every 10% increase in serum BPDE-albumin adduct level in boys, the serum complement C4 level decreased by 1.68% ( P=0.024). After adjusting age, sex and BMI z-score, the levels of serum complement C3 and C4 decreased by 1.31% and 3.57% respectively for every 10% increase in serum BPDE-albumin adducts among children in the urinary cotinine detection group ( P<0.05). Conclusion:There is a significant dose-response relationship between serum BPDE-albumin adducts and the complement C4 among children.
7.Dose-effect relationship between serum polycyclic aromatic hydrocarbon adducts and serum complements among children in a city of East China
Zhenjie WANG ; Yang LIN ; Jingjing YIN ; Liya ZHANG ; Ting WANG ; Yanhua WANG ; Nan WU ; Fanling KONG ; Huawei DUAN
Chinese Journal of Preventive Medicine 2021;55(5):653-659
Objective:To investigate the dose-response relationship between serum polycyclic aromatic hydrocarbon adducts and serum complement C3 and C4 levels among children from a city in East China.Methods:In September 2016, two boarding schools in the air pollution exposure area and the control area (beyond the upwind of 30 km in the air pollution exposure area) in a city in East China were selected as the research site, and the eligible school-age children were recruited as the research objects. A total of 273 children were included, including 163 in the exposure group and 110 in the control group. The annual air pollutant data (PM 2.5, PM 10 and NO 2) of the two regions during the study period were collected. The exposure level of tobacco was evaluated by cotinine in urine. The levels of serum complement C3 and C4 were determined by automatic biochemical analyzer. The serum anti-7, 8, -dihydrodiol-9, 10-epoxide benzo[a]pyrene (BPDE)-albumin adduct levels were detected by ELISA. Linear regression model was used to explore the dose-response relationship between BPDE-albumin adducts and serum complement C3 and C4. Results:The age of 273 subjects was (13.67±0.37) years old, including 165 boys (60.4%). The average annual exposure levels of PM 2.5, PM 10 and NO 2 and the level of serum BPDE-albumin adducts in the exposure group were higher than those in the control group ( P<0.05). The results of linear regression model analysis showed that after adjusting age, sex, BMI z-score and urinary cotinine level, when the serum BPDE-albumin adduct level increased by 10%, the serum complement C4 level decreased by 1.2% ( P=0.017). After adjusting age, BMI z-score and urinary cotinine level, for every 10% increase in serum BPDE-albumin adduct level in boys, the serum complement C4 level decreased by 1.68% ( P=0.024). After adjusting age, sex and BMI z-score, the levels of serum complement C3 and C4 decreased by 1.31% and 3.57% respectively for every 10% increase in serum BPDE-albumin adducts among children in the urinary cotinine detection group ( P<0.05). Conclusion:There is a significant dose-response relationship between serum BPDE-albumin adducts and the complement C4 among children.
8.A preliminary study on the dynamic imaging of chest high resolution CT in patients with mild COVID-19
Zhaoping CHENG ; Yan LI ; Yanhua DUAN ; Hui GU ; Changhu LIANG ; Yumao YAN ; Baojuan GAO ; Shaoxian LI ; Linlin SUN ; Ximing WANG
Chinese Journal of Radiology 2020;54(6):548-551
Objective:To explore the evolution of imaging findings on dynamic chest high resolution CT(HRCT) in patients with mild COVID-19.Methods:The data of epidemiology, clinical data and continuous dynamic chest high-resolution CT images of the patients with mild COVID-19 were analyzed retrospectively. Twenty-five patients (including 24 common type and 1 mild) were enrolled in the group, including 14 males and 11 females, with age of (42±12) years and hospital stay of (19±5) days. The basic images and dynamic images of HRCT were analyzed and compared by the radiologists.Results:The clinical manifestations were fever (22 cases), cough (18 cases), expectoration (8 cases), pharyngeal pain (6 cases). Most laboratory tests lacked specificity. There were no significant abnormalities on chest CT of one mild patient. HRCT findings of the common type were as follows: (1) the distribution of the lesions: most of the multiple lesions involved both lungs (19 cases), with average of 3 (3±1) lobes, located in the peripheral pulmonary zone near the pleura (22 cases); (2) the morphology and density of the lesions: most of the lesions were ground glass density foci (22 cases), which were patchy and massive (18 cases), nodular (10 cases) and arc broadband (7 cases); with the development of the disease, the density of some lesions increased with localized pulmonary consolidation (10 cases), accompanied by air bronchus sign (5 cases) and halo sign (5 cases). Dynamic changes of HRCT images in the chest: the positive manifestations were found on the 5th (5, 6) day after the onset of the disease, the progressive time of CT lesions was 5 (5, 7) days, the peak time of CT lesions was 11 (10, 13) days, and the turning time of CT lesions was 9 (8, 11) days.Conclusions:Dynamic chest HRCT can monitor the basic evolution process of the disease in patients with mild COVID-19, and provide a more intuitive basis for clinical early diagnosis and treatment.
9.Research progress on ViewRay magnetic resonance guided radiotherapy system
Yanchen YING ; Hua CHEN ; Hao WANG ; Hengle GU ; Yanhua DUAN ; Yan SHAO ; Aihui FENG ; Hongxuan LI ; Xiaolong FU ; Zhiyong XU
Chinese Journal of Radiological Medicine and Protection 2019;39(4):316-320
ViewRay magnetic resonance (MR) guided radiotherapy system not only solves the problem of imaging dose,but also can set up accurately,online adaptive radiotherapy and gated irradiation according to magnetic resonance imaging (MRI).The development of this system provides a new technical means of accurate radiotherapy.This review describes the main structure of the ViewRay system,and summarizes quality assurance (QA),dosimetric comparison,respiratory motion management,online adaptive radiotherapy,and preliminary treatment effect.
10.Clinical value of 70 kV combined with low contrast agent CT scan in the diagnosis of pediatric coronary artery disease
Yanhua DUAN ; Li CHEN ; Guangming LU ; Dawei WU ; Zhaoping CHENG ; Ximing WANG
Chinese Journal of Radiology 2018;52(7):507-512
Objective DSA and/or surgery was used as a control to evaluate the image quality, radiation dose and diagnostic efficacy of 70 kV combined with low contrast agent dose and 80 kV dual-source CT scanning of coronary artery. Methods Between Apr, 2012 and Apr, 2017, a total of 150 consecutive pediatrics with suspected or confirmed coronary arterial diseases underwent DSCTCA, and the patients were randomly divided into 3 groups by different scanning protocols. Group A (n=50): 80 kV with 1.5 ml/kg contrast agent;Group B (n=50): 70 kV with 1.5 ml/kg contrast agent;Group C (n=50): 70 kV with 1.0 ml/kg contrast agent. ANOVA was used to compare the differences of subjective coronary arteries image quality between the three groups; The t-test was used to compare the difference in effective radiation dose between the 70 kV group and the 80 kV group; The surgery and/or angiography results were used as gold standard to evaluate the diagnostic efficacy of the three groups. Results All patients underwent DSCTCA successfully and satisfactory diagnostic images were gained . The mean scores of subjective image quality of coronary artery were 3.5±0.7, 3.4±0.6, 3.7±0.8, respectively. There was no significant difference between the 3 groups (F=2.042, P=0.133). The diagnostic accuracy of the three groups for coronary artery disease was 100%, as the surgical and/or angiographic results were used as gold standard. The effective radiation doses in 80 kV group (A group) and the 70 kV group (B+C group) were (0.49 ± 0.04) and (0.30 ± 0.03) mSv, respectively, with a statistically significant difference (t=4.037, P=0.001). Conclusions DSCTCA can better display and diagnose pediatric coronary artery lesions with 70 kV tube voltage combined with low contrast agent, and has a high diagnostic accuracy. It is a reliable method for diagnosing pediatric coronary artery disease.

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