Objective To analyze the micronucleus rate of radiation workers and to provide accurate occupational health monitoring basis in radiation workers exposed to low-level ionizing radiation for a long time. Methods The radiation group consisted of 353 radiation workers who had been exposed to ionizing radiation during work, while the control group consisted of 41 radiation workers who had not yet been exposed to ionizing radiation before work. The cytokinesis-block micronucleus method was used to determine the micronucleus rate. Results The average micronucleus rate in the radiation group was significantly higher than that in the control group (t = −2.95, P < 0.05). In the radiation group, the micronucleus rate gradually increased with age, and the difference was statistically significant (F = 8.36, P < 0.05). The micronucleus rates of workers with > 10 and > 30 years of service were significantly higher than those of workers with < 10 years of service (χ² = −44.79, −60.47, P < 0.05). The micronucleus rate in females was significantly higher than that in males (t = 3.93, P < 0.05). The micronucleus rates in the diagnostic radiology group and the industrial detection group were significantly higher than that in the control group (t = 3.51, 3.65, P < 0.05). Conclusion The micronucleus rate has increased among the radiation workers exposed to low-level ionizing radiation for a long time. It is necessary to further strengthen occupational health monitoring and radiation protection education for radiation workers, especially the medical workers that constitute the largest population of radiation exposure workers.

Objective To investigate the relationship between the expression of matrix metalloproteinase(MMP)-2,MMP-9,MMP-13 and disease activity in pemphigus patients.Methods A total of 60 pemphigus patients treated in the dermatology department of the hospital from January 2021 to January 2023 were select-ed as the study group,and another 60 healthy volunteers who underwent physical examination in the same pe-riod were recruited as the control group.The levels of MMP-2,MMP-9 and MMP-13 in serum and the expres-sion of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells were compared between the two groups,and the relationship between the expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of patients with pemphigus was analyzed.Pemphigus patients were divided into acute phase group(n=22),chronic phase group(n=23)and stable phase group(n=15)according to their disease activi-ty.The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of the three groups were compared,as well as the correlation between the three indexes and disease activity.Receiver operating characteristic(ROC)curve was plotted for analysis.Results The levels of MMP-2,MMP-9 and MMP-13 in serum and the expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells in the study group were higher than those in the control group,with statistical significance(P＜0.05).The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells in severe patients were higher than those in moderate and mild patients,and the difference was statistically significant(P＜0.05).The expression of MMP-2,MMP-9 and MMP-13 and disease activity scores of mononuclear cells in acute attack group were higher than those in chronic attack group,and the differences were statistically significant(P＜0.05).The ex-pression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells were positively correlated with disease activity(rMMP-2=0.545,rMMP-9=0.592,rMMP-13=0.580,P＜0.05).ROC curve analysis showed that compared with the single diagnostic efficacy of MMP-2,MMP-9 and MMP-13 expression in peripheral blood mononuclear cells,the three combined diagnostic efficacy was better.Conclusion The expression of MMP-2,MMP-9 and MMP-13 in peripheral blood mononuclear cells of pemphigus patients is related to the disease activity,and these three indicators can be used as reference indicators for the diagnosis of pemphigus disease activity.

Objective:To study the influence of circular RNA hsa_circZDHHC21_004 on the proliferation of human small intestinal epithelial cells HIEC-6 after 60Co γ-rays exposure. Methods:HIEC-6 cells were exposed to 60Co γ-rays at 0, 5, 10, and 15 Gy with a dose rate of 1 Gy/min. The expression level of hsa_circZDHHC21_004 in the irradiated HIEC-6 cell was detected. Hsa_circZDHHC21_004 was knocked-down to investigate the influences of hsa_circZDHHC21_004 on the proliferation of irradiated HIEC-6 cells by CCK-8 assay and colony formation assay. Results:The expression level of hsa_circZDHHC21_004 in HIEC-6 cells was upregulated by (1.00±0.24), (1.34±0.28), (1.85±0.31), and (2.80±0.64) times of control after 0, 5, 10, and 15 Gy irradiation, respectively and there were significant difference between 10 or 15 Gy group and 0 Gy group ( F=10.86, P=0.008). Knockdown of hsa_circZDHHC21_004 significantly increased the proliferation rate of HIEC-6 cells at 24, 48, and 72 h after 10 Gy irradiation compared with non-irradiated control ( t=-6.25, -5.83, -7.75, P < 0.001). Under 2 and 5 Gy irradiation, the clone formation rates of the hsa_circZDHHC21_004 knockdown cells were significantly higher than those of the control ( t=-7.45, -8.83, P<0.01). Conclusions:Hsa_circZDHHC21_004 is increased after irradiation and influenced the proliferation of irradiated HIEC-6 cells.

Objective To investigate the effects of microRNA497 (miR-497) on the metastasis of gastric cancer and its possible molecular mechanism. Methods SGC-7901 gastric cancer parent cells were cultured in an ultra-low adhesion environment, and the anoikis resistance model of SGC-7901 cells was created after re-adhesion. Clone formation assay, flow cytometry, Transwell^TM test and scratch healing test were used to detect the differences of biological behavior compared with their parent cells. Fluorescence quantitative PCR was performed to detect the expression of miR-497. Western blot analysis was used to detect the changes of key proteins of Wnt/β-catenin signaling pathway and epithelial mesenchymal transformation (EMT) related proteins such as vimentin and E-cadherin. Parent cells and anoikis resistant SGC-7901 cells were transfected with miR-497 inhibitor or miR-497 mimic, and CCK-8 assay was used to detect the proliferation activity. Transwell^TM invasion assay was performed to detect the invasion ability of cells. Transwell^TM migration test and scratch healing assay was used to determine the migration ability. Western blot analysis was used to detect the expressions of Wnt1, β-catenin, vimentin and E-cadherin. By transfecting miR-497 mimic into the anoikis resistance SGC-7901 cells and inoculating them subcutaneously in nude mice, the changes in the volume and mass of tumor tissues were measured and recorded. Western blot analysis was used to determine the expressions of Wnt1, β-catenin, vimentin and E-cadherin of tumor tissues. Results Compared with the parent cells, the anoikis resistance SGC-7901 gastric cancer cells had faster proliferation rate, stronger colony formation, lower apoptosis rate, stronger invasion and migration ability. The expression of miR-497 was significantly decreased. After down-regulation of miR-497, the proliferation ability, invasion and migration ability were significantly enhanced. The expressions of Wnt1, β-catenin and vimentin increased significantly, while E-cadherin decreased notably. The results of up-regulation miR-497 were the opposite. The tumor growth rate, tumor volume and mass of miR-497 overexpression group were significantly lower than those of control group. The expressions of Wnt1, β-catenin and vimentin decreased significantly, while the expression of E-cadherin increased significantly. Conclusion The expression of miR-497 is low in the anoikis resistance SGC-7901 cells. miR-497 can inhibit the growth and metastasis of gastric cancer cells by blocking Wnt/β-catenin signaling pathway and EMT.
Animals ; Mice ; Humans ; beta Catenin/metabolism* ; MicroRNAs/metabolism* ; Vimentin/metabolism* ; Stomach Neoplasms/pathology* ; Anoikis/genetics* ; Wnt Signaling Pathway/genetics* ; Mice, Nude ; Cell Proliferation/genetics* ; Cadherins/genetics* ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition/genetics* ; Cell Movement/genetics*

Objective:To investigate the changes of CPT1A and CPT1B protein expression in rat intestinal epithelial cells (IEC-6) after 60Co γ-ray irradiation, and the mechanism of the influence of carnitine palmitoyltransferase 1 (CPT1) on the proliferation of irradiated IEC-6 cells. Methods:IEC-6 cells were cultured in serum-normal medium or in serum-starved medium overnight, and pretreated with 20 μmol/L palmitic acid (PA) before irradiation with 0, 5, 10, and 15 Gy. At 24 h after irradiation, the cellular protein was collected for the measurement of CPT1A and CPT1B proteins by Western blot. The influences of ETO, an inhibitor of CPT1, on the survival and proliferation of irradiated IEC-6 cells were analyzed by colony formation assay and CCK-8 assay. The protein expressions and phosphorylation levels of the extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK) in 5 Gy irradiated IEC-6 cells pre-treated with ETO were analyzed by Western blot at 48 h after radiation.Results:When IEC-6 cells were cultured in serum-normal medium together with PA, the protein level of CPT1A was significantly increased after 15 Gy irradiation ( t=-2.82, P<0.05). When IEC-6 cells were cultured in serum-starved medium, the protein level of CPT1A was significantly increased at 5, 10, and 15 Gy ( t=-3.28, -8.72, -8.67, P<0.05). When IEC-6 cells were cultured in serum-starved medium together with PA, the protein levels of CPT1A were significantly increased at 5, 10 and 15 Gy ( t=-10.69, -7.02, -8.23, P<0.05), the protein levels of CPT1B were significantly increased at 10 and 15 Gy ( t=-3.73, -5.05, P<0.05). After irradiation, the survival and proliferation of IEC-6 cells in ETO group were significantly lower than those in control group ( t=5.46, 13.22, P<0.05), and the protein level of ERK1/2 and p-JNK in ETO group were significantly lower than those in control group ( t=4.01, 3.29, 10.68, 14.44, P<0.05). Conclusions:CPT1 promoted radiation-induced IEC-6 injury cells survival and proliferation by enhancing the expression level of ERK1/2 protein and the activity of JNK.

Objective:To explore the characteristics of lipid metabolism in rat plasma after total body irradiation(TBI) in order to provide scientific evidence of radiation biomarkers.Methods:For the non-targeted lipidomics study, 50 SD rats were divided into 6 groups and irradiated with 0, 1, 2, 3, 5 or 8 Gy 60Co γ-rays, respectively. For the targeted lipidomics study, 25 rats were divided into 5 groups and irradiated with 0, 0.5, 2.5, 4 or 6 Gy. Venous blood samples were collected and plasma were separated 4 h after TBI. Radiation-sensitive lipids were screened and their concentrations were determined. Receiver operating characteristic curve (ROC) and dose-response were analyzed. Results:A total of 15 radiation differential lipids were screened out based on non-targeted lipidomics study and 7 of them were identified as radiosensitive lipids by targeted lipidomics analysis. The ROC of radiosensitive lipids distinguished area under curve (AUC) of samples in 0 Gy group and > 0 Gy group, < 2 Gy group and ≥ 2 Gy group were all > 0.75. The AUC values were increased to 0.96 and 0.94 after the panel of radiation sensitive lipids ROC analysis. The concentrations of LysoPC(18: 2), LysoPC(22: 0), PC(18: 0/18: 2), PE(18: 2/16: 0) and PE(18: 2/18: 0) decreased with irradiation dose within 0-6 Gy.Conclusions:A total of 7 plasma radiosensitive lipids in rat plasma were identified 4 h after TBI, and the panel of them could be used for specific dose classification. Five of the lipids had good dose-response relationship.

Objective:To investigate the metabolite changes in rat plasma after total body irradiation (TBI) and to explore dose classification based on radiation sensitive metabolites.Methods:The differential metabolites induced by radiation were screened and verified by metabolomics. In the discovery stage, 50 SD rats were irradiated with 0, 1, 2, 3, 5 and 8 Gy of 60Co γ-rays. In the verification stage, 25 rats were irradiated with 0, 0.5, 2.5, 4 and 6 Gy. Peripheral blood samples were collected 4 h after irradiation, and plasma was separated. Radiation-induced differential metabolites were identified and their concentrations were determined. Receiver operating characteristic (ROC) curve of the differential metabolites was used to classify dose range. Results:In the discovery stage, 8 radiation-induced differential metabolites in rat plasma were identified and four of them (cytosine, L-hexylcarnitine, Linoelaidylcarnitine and L-palmitylcarnitine) were upregulated, which was confirmed in the verification stage. The area under the curve (AUC) for the specific dose was >0.75. After combining these four metabolites, the AUC value to classify the radiation dose of 0 Gy versus >0 Gy, <2 Gy versus ≥2 Gy, <5 Gy versus ≥5 Gy were 0.96, 1 and 0.94, respectively.Conclusions:The metabolites in rat plasma changed significantly at 4 h after TBI, where 8 differential metabolites were identified. Cytosine, L-hexylcarnitine, linoelaidylcarnitine and L-palmiylcarnitine were stably over-expressed in the plasma after irradiation. The combination of these four compounds had high classification accuracy and thus may applicable as radiation sensitive biomarkers for dose classification.

Objective To study the diversities of imaging, symptoms, electrophysiology and clinical value of the stereoelectroencephalography(SEEG) in patients with mesial temporal lobe epilepsy.Methods Eight patients with intractable epilepsy in Epilepsy Center of Yuquan Hospital of Tsinghua University who underwent mesial temporal lobectomy were recruited in this study, and their epileptic foci could not be accurately positioned.Therefore stereotactic brain electrodes were implanted, and their usual attack originated from mesial temporal lobe structure were confirmed.There was no seizure in the one year follow-up.Results Symptoms of the eight patients behaved differently, and the onset of the seizures in scalp electroencephalograph or SEEG showed diversities.Epileptic discharges were found originated from the mesial temporal lobe after implanting electrodes: in the early stage of discharges, four cases had the conduction to insular lobe structure;two cases had the conduction to contralateral mesial temporal lobe;one case had the conduction to retrosplenial cortex;one case had the conduction to parietal lobe;one case had the conduction to frontal lobe and rapid generalization (one case had the conduction to insular lobe and contralateral mesial temporal lobe meanwhile).Conclusions There is difference in clinic, imaging and electrophysiology of the patients with mesial temporal lobe epilepsy The non-specificity can be explained by the evolution of the intracranial electroencephalography, which can help us know its network conduction pattern Insular lobe is the most common conduction approach of mesial temporal lobe epilepsy in early stage SEEG can be used as a microinvasive, accurate preoperative localization method, which can help us to locate accurately and understand the discharges and conduction mode.

Objective:To investigate the biological characteristics of monoclonal antibodies against avian influenza virus (AIV).Methods:Monoclonal antibodies (mAbs) against AIV H5N1 were prepared and it′s characteristics were identified including subtype,titer,hemagglutination inhibition activity and cross-reactivity with other influenza viruses.Besides,Western blot and immuno-histochemical staining methods were conducted to test the combination of antibodies and antigen ( H5N1 ) and human normal tissues.Results:Immunohistochemical analysis showed that 2 mAbs (H5-32 and H5-35) cross-reacted with human tissues kidney and pancreas respectively.Conclusion:These data indicated that there have some association between the AIV H 5N1 with human tissues, which may provide reference for the study on avian influenza virus infection and pathogenicity .

Objective To test the immune therapeutic effects of PICKCa adjuvant rabies vaccine for the post-exposure mice and beagle dogs and immune memory effect for human.Methods For the mice and beagle dogs first to infect with rabies wild virus and then immunize with PICKCa rabies vaccine.For volunteers, first to immunize with PICKCa rabies vaccine after 2.5 years to take peripheral blood mononuclear cell and then to check cells secreted interferon-gwith flow cytometry. Results In 3 independent assays of post-explosure immunization of mice and beagle dogs PICKCa rabies vaccine were much better then commercial adjuvant-free rabies vaccines, the protective rate from 20%-30%to 70%-100%Also, in the assay of cells secreted IFN-γby peripheral blood mononuclear cells from volunteers immunized by PICKCa rabies vaccine the IFN-γcell levels were much higher than control.Conclusions Compared with commercial adjuvant-free rabies vaccines the PICKCa rabies vaccine had significant immune therapeutic effects in the mice and dogs.Also PICKCa rabies vaccine had good immune memory effect in human.