1.Inhibitory effect of rosuvastatin on migration and invasion of U87 glioma cells in vitro
Chinese Journal of Comparative Medicine 2018;28(3):52-56
Objective To investigate the inhibitory effect of rosuvastatin on the migration and invasion of U87 glioma cells and its related mechanisms. Methods Cultured U87 cells were treated with 0, 5, 10 and 20 μmol/L rosuvastatin for 24,48 and 72 h. Cell viability was measured by CCK-8 assay. Migration ability of the cells was detected by scratch assay,and invasion ability of the cells was detected by Transwell assay. The expression of matrix metalloproteinase 2(MMP2),MMP9 and the Wnt/β-catenin signaling pathway-related proteins was detected by Western blot assay. Results Compared with the control group,the viability of U87 cells in the 5,10 and 20 μmol/L rosuvastatin groups was decreased (P < 0.01)in a time- and dose-dependent manner. Migration and invasion abilities of the cells were decreased(P <0.01). In addition,the expression levels of MMP2,MMP9,Wnt1,Wnt3a,Wnt7a,and β-catenin were decreased as well (P < 0.01). Conclusions Rosuvastatin can significantly inhibit the migration and invasion abilities of U87 glioma cells, probably related to the blocking of Wnt/β-catenin signaling pathway.
2.Clinical Observation of Sofren Injection Combined with Vinpocetine Injection in the Treatment of Acute Massive Cerebral Infarction
Bing CAO ; Qi DING ; Xipeng LIU ; Chunjiang LIU ; Songhua HU
China Pharmacy 2017;28(32):4527-4529
OBJECTIVE:To investigate clinical efficacy of Sofren injection combined with Vinpocetine injection in the treatment of acute massive cerebral infarction,and its effects on hemorheological indexes and serum NOS.METHODS:A total of 60 patients with acute massive cerebral infarction in our hospital during Jan.2014-Jun.2016 were selected as research objects and divided into trial group and control group according to random number table,with 30 cases in each group.Control group was given Citicoline injection 0.5 g,ivgtt,qd.Trial group was additionally given Vinpocetine injection 20 mg added into 0.9% Sodium chloride injection 250 mL,ivgtt,qd;1 h later washing tube,they were given Sofren injection 10 mL added into 0.9% Sodium chloride injection 250 mL,ivgtt,for consecutive 14 d.Clinical efficacies and safety of 2 groups were observed,and hemorheological indexes and NOS levels were observed before and after treatment.RESULTS:The total response rate (83.33%)of trial group was significantly higher than that (50.00%) of control group,with statistical significance (P<0.05).Before treatment,there was no statistical significance in hemorheological indexes or serum NOS levels between 2 groups (P>0.05).After treatment,hemorheological indexes of 2 groups were decreased significantly,and the trial group was significantly lower than the control group.The level of serum NOS in 2 groups were increased significantly,and the trial group was significantly higher than the control group,with statistical significance (P<0.05).No obvious ADR was found in 2 groups.CONCLUSIONS:Sofren injection combined with Vinpocetine injection show significant therapeutic efficacy for acute massive cerebral infarction,can reduce blood viscosity and increase blood perfusion with good safety.
3. An analysis of assessment results of prevention and control of occupational hazards in a city
Lin YANG ; Xipeng SU ; Fei YE ; Wei ZHANG ; Yang XING ; Hui LI ; Zheng JI ; Qian LIU ; Bingbing LIU ; Jie NIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(12):899-903
Objective:
To investigate the current status of prevention and control of occupational hazards in a city in 2017, to understand the capability to prevent and control occupational hazards and the level of occupational health supervision and management, and to propose measures to urge employers to assume the main responsibility for the prevention and control of occupational diseases.
Methods:
An analysis of the main factors influencing the prevention and control of occupational hazards in the city was performed to screen out six semi-quantitative assessment indicators (including the training of the main responsible persons and occupational health management personnel in companies) and four qualitative assessment indicators (including the coverage of supervision and inspection of occupational hazards performed by the district safety supervision department) , which could be used to measure the prevention and control effects of regional occupational hazards. Each indicator was scored. The typical investigation method was used to do data review and on-site inspection of 170 companies, 17 district-level occupational health supervision departments, and 16 sub-district (township) occupational health supervision departments in the city from October to December, 2017. The prevention and control of occupational diseases in each district was scored, and the completion rate and completion situation of each indicator in the city were analyzed.
Results:
The mean score of prevention and control of occupational hazards in the city was 84.9. The scores of two districts were relatively high, being 88.9 and 88.7, respectively; the scores of 9 (52.9%) districts were higher than 85. The pass rate of training for the main responsible persons and occupational health management personnel in companies in the city was 95.9%, the pass rate of occupational health training for workers was 84.7%, the pass rate of occupational health examination for workers was 96.5%, the pass rate for the inspection of occupational hazardous factors in workplaces was 95.3%, the pass rate for notifying occupational hazards in workplaces is 95.9%, and the pass rate for applying the warning signs of occupational hazards in workplaces was 76.5%.
Conclusion
The awareness of the importance of prevention and control of occupational hazards in all districts of the city has been increased, but the effectiveness of occupational health training for workers needs to be strengthened. The supply and demand of occupational health technical services are still not matched, and the ways and methods of occupational health supervision and inspection need to be improved.
4.Effect of diosmetin on ferroptosis of hippocampal cells in rats with bacterial meningitis and the mechanism of SIRT1-Nrf2 signaling pathway
Lu ZHANG ; Ziting WANG ; Xipeng LIU ; Xiufeng ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(7):584-591
Objective:To explore the effect and mechanism of diosmetin (Dio) on neuronal ferroptosis in rats with bacterial meningitis (BM).Methods:Male SD rats aged 6-7 weeks of SPF grade were selected for the experiment. The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum. Sixty BM model rats were successfully modeled and divided into model group, low-dose Dio group, medium-dose Dio group, high-dose Dio group and inhibitor group according to the random number table method, with 12 rats in each group. Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group, medium-dose Dio group, high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg, 100 mg/kg, 200 mg/kg and 200 mg/kg, respectively. The rats in the control group were intragastrically administered with an equal volume of 0.9 % sodium chloride solution. On the day of intragastric administration, the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527 (10 mg/kg), and the rats in the other groups were injected with an equal volume of 0.9% sodium chloride solution. The above interventions were performed once a day for 28 consecutive days. Loeffler neurological score was used to evaluate the neurological impairment in rats. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cerebrospinal fluid of rats were detected by ELISA. The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer. Glutathione (GSH) was detected by micro-enzyme labeling method, malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, reactive oxygen species(ROS) was detected by colorimetry, and Fe 2+ level was detected by ferrozine method. Hematoxylin-eosin staining, Prussian blue staining and TUNEL staining were used to observe the pathological damage, iron accumulation and apoptosis in the hippocampus, respectively.Western blot was applied to measure the expression of transferrin (Tf), proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), caspase-3 and SIRT1/Nrf2/HO-1/Gpx4 signaling pathway proteins. Graphpad Prism 9.0 was used for data analysis. One-way ANOVA was used for statistical analysis, and SNK- q test was used for further pairwise comparisons. Results:(1) There was a statistically significant difference in neurological function scores among the 6 groups of rats ( F=125.451, P<0.001). The neurological function score of the model group was lower than that of control group, while the neurological function scores of the low-dose Dio group, medium-dose Dio group, and high-dose Dio group were higher than those of the model group (all P<0.05). The neurological function score of the inhibitor group ((2.57±0.26)) was lower than that of high-dose Dio group ((4.34±0.48)) ( P<0.05). (2) There were statistically significant differences in the levels of IL-6, TNF-α and the number of white blood cells in the cerebrospinal fluid of rats among the 6 groups ( F=127.817, 102.413, 180.967, all P<0.001). The levels of IL-6, TNF-α and the number of white blood cells in model group were higher than those of control group(all P<0.05). The levels of IL-6, TNF-α and the number of white blood cells in low-dose Dio group, medium-dose Dio group and high-dose Dio group were lower than those of model group (all P<0.001), and those in inhibitor group were all higher than those in high-dose Dio group(all P<0.001). (3) There were statistically significant differences in iron deposition rate and neuronal apoptosis rate among the 6 groups of rats ( F=90.857, 88.835, both P<0.001). The iron deposition rate ((18.37±3.14)%) and neuronal apoptosis rate ((27.58±2.63)%) in the inhibitor group were higher than those in the high-dose Dio group ((6.35±1.08)%, (14.02±1.87)%) (both P<0.05). (4) The levels of GSH, ROS, MDA, and Fe 2+ in the hippocampus of the 6 groups of rats showed statistically significant differences ( F=54.465, 106.453, 55.969, 105.457, all P<0.001). The GSH content in the inhibitor group ((103.48±8.76) mmol/g) was lower than that in the high-dose Dio group ((133.97±10.54) mmol/g), while the contents of ROS, MDA, Fe 2+ ((225.17±16.32) μmol/mg, (10.73±1.58) μmol/mg, (62.71±5.43) μg/g) were higher than those of the high-dose Dio group ((131.87±11.67) μmol/mg, (4.35±0.87) μmol/mg, (34.86±2.95) μg/g) (all P<0.05). (5)There were statistically significant differences in the protein levels of Tf, PCNA, Bax, caspase-3, SIRT1, Nrf2, HO-1 and Gpx4 in the hippocampus of the 6 groups of rats ( F=120.179, 107.568, 157.265, 98.031, 90.932, 52.283, 59.424, 114.539, all P<0.001). The protein levels of Tf, Bax and caspase-3 in the hippocampus of inhibitor group were higher than those of the high-dose Dio group, while the protein levels of PCNA, SIRT1, Nrf2, HO-1, Gpx4 were lower than those of the high-dose Dio group (all P<0.05). Conclusion:Diosmetin can activate SIRT1/Nrf2/HO-1/Gpx4 signaling pathway, thereby inhibiting neuronal ferroptosis in BM rats.
5.Mechanism of silymarin on malignant growth of glioma cells by regulating miR-124-3p/WEE1 axis
Ming LIU ; Xipeng LIU ; Chun LI ; Xiufeng ZHANG ; Bing CAO ; Jianxin QIAO ; Xue WANG
Journal of China Medical University 2024;53(2):142-148
Objective To investigate the impact of silymarin(SM)on the malignant growth of glioma cells and the regulatory mechanism on the miR-124-3p/WEE1 axis.Methods Glioma U87 cells were grouped into control,SM low,medium,and high concentration groups,and SM high concentration + miR-124-3p inhibitor group(SM high + miR-124-3p inhibitor group).CCK-8 was used to measure the proli-feration rate of cells;Transwell? assay was applied to assay the migration and invasion of cells;cell cycle progression was detected by flow cytometry;Western blotting was applied to measure the expression of cyclin D1 and apoptosis-related proteins;the levels of miR-124-3p and WEE1 mRNA were determined by qRT-PCR;and a luciferase activity test was applied to verify the targeting relationship between miR-124-3p and WEE1;in addition,the establishment,administration,and analysis of a NOD/SCID mouse model of intracranial trans-planted tumor were conducted.Results Compared with the control group,the cell proliferation,the numbers of migrating and invading cells,the expression of cyclin D1,and the level of WEE1 mRNA in the various SM treatment groups decreased,the number of cells in G0/G1 phase,the expression of cleaved caspase-8,cleaved caspase-9,cleaved caspase-3 and miR-124-3p increased(P<0.05);furthermore,transfection of miR-124-3p inhibitor reversed the inhibitory effect of SM on the malignant behavior of glioma cells.In vivo experiments with mice showed that the weights and volumes of tumors in the SM treatment group were lower than those in the model group(P<0.05),and there was no discernible change in the weight of the mice(P>0.05).Conclusion SM can inhibit the malignant growth of glioma cells by upregulating miR-124-3p and downregulating WEE1.
6.Relation of genetic polymorphism of NQO1 and GSTT1 with risks of chronic benzene poisoning.
Junxiang WAN ; Jinxiu SHI ; Jiru GUAN ; Rong YE ; Xiaoling GAO ; Weiwei LIU ; Lijian HUI ; Duozhi CAO ; Xipeng JIN ; Gengxi HU ; Zhaolin XIA
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(5):340-343
OBJECTIVETo explore the relation between genetic polymorphisms of NQO1, GSTT1 and risks of chronic benzene poisoning (BP).
METHODSA case-control study was conducted. 152 BP patients and 152 workers occupationally exposed to benzene without poisoning manifestations were investigated. Polymerase chain reaction (PCR), denaturing high-performance liquid chromatography(DHPLC) and sequencing were used to detect the single nucleotide polymorphisms(SNPs) of the promoter and complete coding-region of NQO1 gene. Multiple PCR was used to detect GSTT1 genotype.
RESULTSIn smoking population, there was 7.73-fold (95% CI: 1.71-34.97, P = 0.010) of risk in BP subjects carrying NQO1c. 609 T/T genotype, compared with those carrying C/C and C/T. genotype. In drinking population, the individuals carrying the 6th extron of NQO1c. 609 T/T homozygote genotype had a 11.00-fold(95% CI: 1.89-63.83, P = 0.005) risk of BP compared to those with NQO1c. 609 C/T and C/C genotypes.
CONCLUSIONThe subjects carrying NQO1c. 609 T/T genotype and together with the habit of smoking or drinking may be more susceptible to BP.
Benzene ; poisoning ; Case-Control Studies ; Ethanol ; adverse effects ; Genotype ; Glutathione Transferase ; genetics ; Humans ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; Occupational Diseases ; genetics ; Occupational Exposure ; Polymorphism, Single Nucleotide ; Smoking ; adverse effects
7.Clinicopathological characteristics of colorectal cancer complicated with type 2 diabetes mellitus: analysis of clinicopathological data from 3, 202 colorectal cancer patients.
Rui LIU ; Liling HU ; Gang LI ; Lizhong ZHAO ; Sha LI ; Xipeng ZHANG ; Qinghuai ZHANG ; Lina WU
Chinese Journal of Oncology 2014;36(1):74-77
OBJECTIVEthe aim of this study was to determine the clinicopathological characteristics of colorectal cancer (CRC) patients complicated with type 2 diabetes mellitus (T2DM ).
METHODSA total of 3, 202 patients with CRC confirmed pathologically in Tianjin Union Medicine Center from January 2005 to December 2009 were included in this study. We analyzed the differences in clinicopathological features between T2DM patients and non-diabetic patients according to age of diagnosis, gender, tumor site, stage, gross type, histological type, and differentiation.
RESULTSFrom 2005 to 2009, the number of CRC patients increased yearly. The high incidence age of all CRC patients was 51 to 80 years old. The male to female ratio was 1.18:1, showing that the number of female patients with CRC was increased significantly compared with males. The CRC distribution of T2DM patients and non-diabetic patients showed a predominance of rectal cancer (64.4%, 68.7%), followed by sigmoid colon cancer (12.5%, 13.0%), and moderately differentiated ulcer-type adenocarcinoma. Compared with non-diabetic patients, T2DM patients were older (66.2 years versus 62.7 years, P < 0.001) and had more multiple CRCs (3.5% versus 1.6%, P < 0.001). Moreover, the proportion of lymph node or organ metastasis in T2DM patients was higher than that in non-diabetic patients (52.6% versus 45.6%, P < 0.05). No significant differences were observed between both groups in terms of gender, gross type, histological type, and differentiation(P > 0.05 for all).
CONCLUSIONSCRC incidence shows an increasing trend with age. CRC patients with T2DM have an older age of onset, higher proportion of lymph node and distant organ metastasis than in non-diabetic patients.
Adenocarcinoma ; complications ; pathology ; Aged ; Colorectal Neoplasms ; complications ; pathology ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Male ; Middle Aged
8.Characterization of a family B DNA polymerase from Thermococcus eurythermalis A501 and its application in PCR.
Chinese Journal of Biotechnology 2022;38(2):807-819
DNA polymerases are widely used in PCR and play important roles in life science research and related fields. Development of high-performance DNA polymerases is of great commercial interest as the current commercial DNA polymerases could not fully satisfy the requirements of scientific research. In this study, we cloned and expressed a family B DNA polymerase (NCBI accession number TEU_RS04875) from Thermococcus eurythermalis A501, characterized its enzymatic property and evaluated its application in PCR. The recombinant Teu-PolB was expressed in E. coli and purified with affinity chromatography and ion-exchange chromatography. The enzymatic properties of Teu-PolB were characterized using fluorescence-labeled oligonucleotides as substrates. The application potential of Teu-PolB in PCR was evaluated using the phage λ genomic DNA as a template. Teu-PolB has DNA polymerase and 3'→5' exonuclease activities, and is highly thermostable with a half-life of 2 h at 98 ℃. The most suitable PCR buffer is consisted of 50 mmol/L Tris-HCl pH 8.0, 2.5 mmol/L MgCl2, 60 mmol/L KCl, 10 mmol/L (NH4)2SO4, 0.015% Triton X-100 and 0.01% BSA, and the optimal extension temperature is 68 ℃. Under the optimized conditions, a 4 kb target fragment was successfully amplified with an extension rate of 2 kb/min. The yield of the Teu-PolB amplified-DNA was lower than that of Taq DNA polymerase, but its extension rate and fidelity was higher than that of Taq and Pfu DNA polymerases. The biochemical properties of Teu-PolB demonstrate that this enzyme can be used in PCR amplification with high thermostability, good salt tolerance, high extension rate and high fidelity.
DNA-Directed DNA Polymerase/genetics*
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Escherichia coli/genetics*
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Polymerase Chain Reaction/methods*
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Temperature
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Thermococcus/genetics*
9.Therapeutic efficacy and mechanism of dandelion extract on intracerebral hemorrhage rats
Xiufeng ZHANG ; Xiaofei LI ; Jinghui WANG ; Huijing WANG ; Liwen WANG ; Cheng ZHEN ; Pengfei WANG ; Xipeng LIU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(2):212-216
Objective To investigate the therapeutic efficacy of dandelion extract on intracerebral hemorrhage(ICH)rats and its effect on nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods Stereotaxic intracranial injection of type Ⅳ col-lagenase was used to establish rat ICH model.Then 48 ICH rats were randomly divided into mod-el group,dandelion extract group,Nrf2 inhibitor(ML385)group and dandelion extract+ML385 group,with 12 rats in each group.Another 12 rats served as sham operation group.After treat-ment,neurological deficits was evaluated and scored for all groups of rats.Blood-brain barrier(BBB)function,neuronal apoptotic rate in the hippocampus,serum levels of COX-2,IL-6 and iNOS,cerebral contents of CAT,GSH-Px,ROS and MAD,and protein levels of Nrf2/HO-1 signal pathway were detected.Results Compared with sham operation group,the neurological deficit score,Evans blue exudation,appptotic rate of hippocampal neurons,serum COX-2,IL-6,iNOS levels,brain tissue reactive oxygen species(ROS)and malondialdehyde level in the model group were significantly increased(P<0.05),and the expression levels of CAT,GSH-Px,Nrf2 and HO-1 proteins were significantly decreased(P<0.05).Compared with dandelion extract group,combination of dandelion extract and ML385 significantly increased the neurological deficit score(2.54±0.23 vs 1.43±0.19),Evans blue exudation[(22.15±3.61)ng/mg vs(6.54±1.24)ng/mg],apoptotic rate[(31.97±5.26)%vs(3.51±0.94)%],serum COX-2[(5.82±1.16)ng/ml vs(1.34±0.42)ng/ml],IL-6[(1.47±0.31)ng/ml vs(0.43±0.14)ng/ml]and iNOS levels[(59.91±10.36)U/ml vs(13.94±3.78)U/ml],brain tissue ROS[(4.70±0.45)U/kg vs(1.70± 0.51)U/kg]and MDA levels[(3.72±0.52)nmol/mg vs(1.17±0.34)nmol/mg],and decreased expression levels of CAT[(2.54±0.59)U/mg vs(5.68±1.04)U/mg],GSH-Px[(8.01±0.86)U/mg vs(16.97±3.03)U/mg],Nrf2(0.67±0.13 vs 1.07±0.19)and HO-1(0.55±0.07 vs 0.86± 0.10,P<0.05).Conclusion Dandelion extract can enhance the antioxidant activity in ICH rats by activating Nrf2/HO-1 signaling pathway,prevent the progression of inflammation and oxida-tive stress,inhibit neuronal apoptosis in hippocampus,repair blood-brain barrier function,and thus improve nerve function.
10.Analysis of occupational radiation sicknesses diagnosed in China from 2013 to 2017
Xiaoliang LI ; Yinping SU ; Shujie LEI ; Xipeng ZHAO ; Yinghua FU ; Shuxia HAO ; Jianxiang LIU ; Quanfu SUN
Chinese Journal of Radiological Medicine and Protection 2018;38(10):779-783
Objective To analyze the changes in distribution of occupational radiation cases reported from 2013 to 2017 in China and learn about the occupational health risks of radiation workers.Methods Descriptive analyses were made of regional distribution,disease category distribution,occupation category distribution and exposure mode distribution of these cases,according to the reports (2013-2017) of occupational radiation sickness from " Occupational Health of Radiation Workers Management System".Results There were 54 diagnostic radiology agencies for occupational radiation sickness in China that covered all provinces,autonomous regions and municipalities except Tibet and Production and Construction Corps of Xinjiang.A total of 106 new cases were reported from 2013 to 2017.Most of the cases were radiogenic neoplasm (43.40%),and chronic radiation sickness were from external exposure (16.98%) and radiation cataract (16.04%).Most of the cases (70.75%) were engaged in medical application and a small part of the cases (13.21%) engaged in industry application.Chronic exposure (80.19%) was the most frequent form of exposure mode,but acute exposure (5.66%) was very few.A part of cases (14.15%) were reported without exposure mode.Conclusions The morbidity of occupational radiation sickness declined generally in China and occupational health management of key workers should be strengthened continuously.