1.To Investigate the Progression Mechanism of Breast Cancer in Mice Induced by Chronic Restraint Stress and the Regulatory Mechanism of Xiaoyaosan Based on TGF-β1/CD147 Signal
Yan LIU ; Gaoyuan LIU ; Mengwei PENG ; Luoyi KUANG ; Zilong LUO ; Tiezhu YANG ; Yaosong WU ; Yulong CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(4):934-943
Objective To investigate the progression of breast cancer in mice induced by chronic binding stress and the regulatory mechanism of Xiaoyao SAN based on TGF-β1/CD147 signal pathway.Methods 40 BABL/c mice were randomly divided into tumor group,model group,Xiaoyaosan group and Mifepristone group,and then 4T1 cell line was inoculated into the armpits of each group of mice.After Tumor formation,mice in all groups except tumor group were subjected to chronic restraint stress for 21 days.Meanwhile,mice in Xiaoyaosan and Mifepristone groups were gavaged with the corresponding drugs,and mice in the other two groups were gavaged with normal saline.After the modeling,the mice were sacrificed after anaesthesia.The weight and volume of the tumors and visceral index of the mice were measured.The contents of serum tumor markers(CA199,CEA,VEGF),serum neurotransmitters(DA and CORT),and inflammatory mediators(TGF-β1 and IL-10)in tumor tissues were detected by Elisa.The expressions of iNOS and Arg-1,the polarization markers of macrophages,and the expressions of CD147 and its downstream signaling molecules MMP2,MMP9 and VEGF in tumor tissues were all detected by immunohistochemistry and Western blot.Results Compared with tumor group,in model group,tumor weight and volume,serum CA199,CEA,VEGF,CORT content,tumor TGF-β1 and IL-10 content were significantly increased;visceral index and serum DA content were significantly reduced;the expression of M2-type polarization marker Arg-1 in tumor macrophages was significantly increased,while the expression of M1-type polarization marker iNOS was significantly decreased;the expressions of CD147 and its downstream signaling molecules MMP2,MMP9 and VEGF were significantly increased.Both Xiaoyaosan and mifepristone could effectively reverse the above changes.Conclusion The mechanism of chronic restraint stress promoting breast cancer progression in mice is related to the increased release of TGF-β1 from M2-type polarization of tumor-associated macrophages,which activates CD147 and its downstream related signals.Xiaoyaosan could relieve the M2-type polarization of macrophages caused by increased corticosterone under stress conditions,reduce the production of TGF-β1,inhibit CD147 and its downstream signal,and thus inhibit the progression of breast cancer caused by chronic restraint stress in mice.
2.Epidemiological characteristics of severe fever with thrombocytopenia syndrome in China, 2018-2021
Xiaoxia HUANG ; Shanshan DU ; Aqian LI ; Chuan LI ; Tingting TIAN ; Tiezhu LIU ; Qin WANG ; Mifang LIANG ; Shiwen WANG ; Jiandong LI ; Dexin LI
Chinese Journal of Epidemiology 2024;45(1):112-116
Objective:To understand the epidemiological characteristics and incidence trend of severe fever with thrombocytopenia syndrome (SFTS) in China.Methods:The incidence data of SFTS in China from 2018 to 2021 were collected from Chinese Disease Prevention and Control Information System for a statistical and descriptive epidemiological analysis by using software such as Excel 2016, Joinpoint 5.0.2, SPSS 26.0, and GraphPad Prism 8.0, especially, the SFTS cases reported monthly by key provinces were analyzed.Results:From 2018 to 2021, a total of 8 835 SFTS cases were reported in 25 provinces and the annual incidence showed an upward trend. The distribution of SFTS cases showed clustering, but the cases were mainly sporadic ones. The cases began to increase in March, mainly occurred during April to October (96.79%,8 551/8 835), and peaked during May to July. The cases were mainly distributed in middle-aged and old farmers, and slight more cases were women. The average case fatality rate was 5.38%, which varied greatly with areas. The case fatality rate tended to increase with age.Conclusion:From 2018 to 2021, the epidemiological characteristics of SFTS in China remained stable, but the number of reported cases gradually increased and the distribution showed an expanding trend, to which close attention should be paid.
3.Exploration of mouse model of breast cancer complicated with depression induced using 4T1 breast cancer cell inoculation combined with chronic restraint stress
Yan LIU ; Mengwei PENG ; Gaoyuan LIU ; Tiezhu YANG ; Wenxian ZHANG ; Yaosong WU ; Yulong CHEN
Chinese Journal of Comparative Medicine 2024;34(3):36-44,50
Objective To study the core behavioral symptoms,biological indicators,and pathological changes of a mouse model of breast cancer complicated with depression induced using 4T1 breast cancer cell inoculation combined with chronic restraint stress(CRS).Methods BABL/c mice were randomly divided into Control,Stress,Tumor,and stress combined with tumor(S+T)groups.Mice in the tumor and S+T groups were inoculated under the front legs with breast cancer 4T1 cells.After tumor formation,mice in the stress and S+T groups were subjected to CRS for 21 days.The body weight and food intake of each group were monitored during modeling.After the experiment,the occurrence of depression-like behavior of mice in each group was evaluated by sucrose preference test,open field test,elevated plus-maze test,and forced swimming test.After the mice were decapitated,the weights and volumes of the tumors were measured.Concentrations of serum tumor markers,including carbohydrate antigen(CA199),carcinoembryonic antigen(CEA),and vascular endothelial growth factor(VEGF),and related neurotransmitters,including 5-hydroxytryptamine(5-HT),norepinephrine(NE),and corticosterone(CORT),were determined using ELISA.HE staining was used to observe histopathological changes to the hippocampus and tumor.Results In S+T group mice,body weight and food intake were significantly decreased,tumor weight and volume were significantly increased,serum tumor marker(CA199,CEA,VEGF)levels were significantly increased,enthusiasm and desire to explore a new environment were reduced,stress and despair behaviors were significantly increased,and levels of the serum neurotransmitters 5-HT and NE and levels of CORT were significantly increased.In addition,the cell arrangement in the tumor tissue was loose,the amount of intercellular substance decreased,the pathological nuclear classification phase was increased,the arrangement and morphology of neurons in the CA3 region of the hippocampus were disordered,and there were obvious nuclear vacuolation-like changes.Conclusions A mouse model of breast cancer complicated with depression induced by 4T1 breast cancer cell inoculation combined with CRS showed the typical dual symptoms and biological indicators of breast cancer and depression and can be used as a good reference model for experimental studies of breast cancer complicated with depression.
4.Preliminary investigation and risk analysis of the infection with lymphocytic choriomeningitis virus and hantavirus in 139 cross-border migrant workers
Hao LI ; Meijun GUO ; Xiaoxia HUANG ; Shanshan DU ; Wei WU ; Aqian LI ; Qin WANG ; Chuan LI ; Tiezhu LIU ; Tingting TIAN ; Lina SUN ; Shiwen WANG ; Mifang LIANG ; Dexin LI ; Jiandong LI
Chinese Journal of Experimental and Clinical Virology 2023;37(1):45-49
Objective:To investigate the prevalence of lymphocytic choriomeningitis virus (LCMV) and hantavirus (HV) specific antibodies among cross-border migrant workers for assessment of the risk of rodents-borne virus infection.Methods:From 2019 to 2020, a survey was conducted on cross border migrant workers engaged in outdoor activities, and serum samples were collected, LCMV specific IgG antibody was detected by an indirect ELISA and Western blot based on recombinant nucleoprotein, and indirect immunofluorescence assay (IFA) based on recombinant expressed glycoprotein. HV IgG antibody in serum was detected by a commercial indirect IgG ELISA kit and IFA based on hantavirus infected Vero cells.Results:A total of 139 cross-border workers, aged 25~57, were surveyed; 64% (89/139) had working experience in multiple countries, involving 26 countries, including 14 countries in Asia and 12 countries in Africa; 11.51% (16/139) of serum samples were tested positive for LCMV antibodies, and the positive samples were verified by Western blot and IFA. The antibody detection rate was slightly higher than the published infection rate from other similar studies. And, HV antibodies were detected from one serum sample (0.72%, 1/139) by ELISA and IFA. However, it was still uncertain when and where the viral infections were acquired.Conclusions:Through this serological cross-sectional preliminary analysis, the infection status and existing risks of LCMV and HV viruses among cross border migrant workers were revealed, which suggested the necessity of strengthening the prevention and control of rodents borne diseases in outdoor engineering sites.
5.Monochromatic images based on spectral CT for stent evaluation after transjugular intrahepatic portosystemic shunt
Jiawei WANG ; Tiezhu LI ; Xiaohui ZHANG ; Pingping WANG ; Bin FENG ; Zhou LIU ; Ping CHEN ; Donghong WEI ; Peng YANG ; Haitao ZHAO ; Baoying CHEN
Chinese Journal of Radiology 2022;56(6):673-677
Objective:To investigate the optimal monochromatic level for evaluation of in-stent lumen after transjugular intrahepatic portosystemic shunt (TIPS) by dual-layer detector CT.Methods:Twenty-nine patients after TIPS were retrospectively enrolled who underwent abdomen enhanced examinations with portal venous phases by a dual-layer detector CT between December 2019 and July 2021. The mixed iterative image (conventional group) and monochromatic images (40 keV group, 50 keV group, 60 keV group and 70 keV group) were obtained by reconstruction. Circular regions of interest were placed in the in-stent of the cross-sectional reconstructed image and in the vertical spinal muscle on the same plane to obtain the corresponding average CT value and noise. The contrast to noise ratio (CNR) and signal to noise ratio (SNR) were calculated. Then 4-point scale was performed to evaluate image quality subjectively by 2 physicians blindly and separately. One-way ANOVA or Kruskal Wallis H rank-sum test was used for the overall analysis between groups, and LSD test or Dunn′s Bonforoni test was used for pairwise comparison within groups. Results:There was no significant difference in noise values among the 5 groups ( P>0.05). The difference of CNR and SNR between the 5 groups was statistically significant ( F=72.28, 56.45, P<0.001). The CNR and SNR in the 40 keV group were the highest, which were 50.4±15.7 and 59.3±18.4 respectively, and the difference was statistically significant ( P<0.001). Subjective scores showed statistically significant differences among the 5 groups (χ2=101.61, P<0.001). The score of the 40 keV group was higher than that of the 60 keV group, 70 keV group, and conventional group ( P<0.001), and there was no significant difference when compared with the subjective score of the 50 keV group ( P>0.05). Conclusions:The 40 keV monochromatic image of dual detector spectral CT is the best image to observe the lumen of the stent after TIPS.
6.Research progress in life cycle and antiviral drugs for severe fever with thrombocytopenia syndrome virus
Tiezhu LIU ; Mifang LIANG ; Shiwen WANG
Chinese Journal of Experimental and Clinical Virology 2022;36(4):475-480
Severe fever with thrombocytopenia syndrome virus (SFTSV), is a tick-borne novel Bunyavirus, which was first discovered in the area of Dabie Mountain in China. It can cause severe fever and thrombocytopenia clinically, with a fatality rate of 10% to 30%. As the pathogenesis and infection mechanism of SFTSV are still unclear, there are no specific drugs and vaccines for SFTSV. In this paper, the mechanism of entry, replication, assembly and budding of SFTSV is reviewed, and the research progress of the treatment antibody and antiviral drugs of SFTSV is summarized, which will provide reference for the development of vaccines and antiviral drugs for SFTSV in China.
7.Determination of three constituents in compound ketoconazole ointment by RP-HPLC
Bo WU ; Tiezhu WANG ; Nian BAI ; Bing LIU ; Hongtao SONG
Journal of Pharmaceutical Practice 2021;39(2):152-156
Objective To establish a RP-HPLC method for determination of ketoconazole, mupirocin and mometasone furoate in compound ketoconazole ointment. Methods RP-HPLC was conducted on a Intersil ODS-3 column (250 mm×4.6 mm, 5 μm), with methanol-PBS with pH 5.5 (65:35) as the mobile phase and the column temperature was 45 ℃. The flow rate was 1.0 ml/min, and the detection wavelength was 248 nm. Results The methodological verification showed that ketoconazole, mupirocin and mometasone furoate had a good linearity (r≥0.9995). The inter/intra-day precisions were less than 3.0%, The recovery rates were between 90% and 108%. The stability and repeatability of RSD were also less than 3.0%, which met the requirements of method validation. The contents of the three components in three batches were determined by the new method. Conclusion The method is simple and reliable. It can provide a basis for the quality control of compound ketoconazole ointment and lay a foundation for its quality standard research.
8. Methods of packaging lentivirus library for CRISPR/cas9 -based screening
Tiezhu LIU ; Aqian LI ; Naizhe LI ; Yuanyuan QU ; Chuan LI ; Quanfu ZHANG ; Yang LIU ; Dexin LI ; Mifang LIANG ; Shiwen WANG
Chinese Journal of Experimental and Clinical Virology 2019;33(2):207-211
Objective:
To obtain the optimum of lentiviral library packaging based on CRISPR/cas9 (Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9).
Methods:
Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence antibody (IFA) and enzyme linked immunosorbent assay (ELISA) were used to detect the lentivirus titers in condition of different ratio of packaging plasmids, different addition of lipofectamine 3000 reagent and different time points post-transfection. Then, high-throughput sequencing was performed to evaluate the representation and distribution of single guide (sg)RNAs in the library.
Results:
The lentivirus titer was the highest when the molar ratio of psPAX2∶pMD2.0G∶Lentivirus library was 2∶1∶1, and the optimum addition of Lipofectamine 3000 reagent was 10 μl, while the result of ELISA were correspondent to that of RT-PCR. The IFA result showed that the lentivirus titer was the highest at 60 h post-transfecion. The coverage of sgRNAs in the lentivirus library packaged with the optimum we obtained was 99.3%, and the read counts of sgRNAs was observed in a normal distribution.
Conclusions
The optimal lentivirus library packaging was obtained, and this can provide basis for CRISPR/cas9-based screening.
9.Whole-sequence Analyses for 12 HBV C/D Recombinants from a Population in Tibet (China).
Tiezhu LIU ; Liping SHEN ; Wenjiao YIN ; Feng WANG ; Fuzhen WANG ; Guomin ZHANG ; Hui ZHENG ; Duoji DUNZHU ; Shengli BI ; Fuqiang CUI
Chinese Journal of Virology 2016;32(2):156-160
We wished to undertake molecular genetic typing and evaluate recombinants of the hepatitis-B virus (HBV) in Tibet (China). Multistage random sampling was used to collect HBsAg-positive samples. Nested polymerase chain reactions were used to amplify the whole sequence of the HBV. DNAstar, MEGA6 and SimPlot were used to assemble sequences, create phylogenetic trees, and undertake recombination analyses. Twelve whole sequences of the HBV of a Tibetan population were collected using these methods. Results showed that all 12 strains were C/D recombinants. Nine of the recombinations were at nt750, and the other three at nt1526. Therefore, the 12 strains could be divided into two types of recombinants: C/Da and C/Db. Analyses of the sequence of the whole genome revealed that the 12 strains belonged to genotype C, and that the nucleotide distance was > 4% between the 12 strains and sub-genotypes C1 to C15 in Genbank. The most likely sub-genotype was C1. Individuals with C/Da were from central and northern Tibet (e.g., Lasa, Linzhi, Ali) and those with C/Db recombinants were from Shannan in southern Tibet. These data suggest that the two types of recombinants had a good distribution in Tibet. Also, they can provide important information for studies on HBV recombination, gene features, virus evolution, as well as the control and prevention of HBV infection in Tibet.
Adult
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Female
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Genotype
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Hepatitis B
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virology
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Hepatitis B virus
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classification
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genetics
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isolation & purification
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Humans
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Male
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Phylogeny
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Recombination, Genetic
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Sequence Analysis, DNA
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Tibet
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Young Adult
10.Human enhancement technology and its military applications and impacts
Tiezhu LOU ; Shu LIU ; Tianxi DIAO
Military Medical Sciences 2014;(1):6-9
Human enhancement technology is an emerging field that aims to enhance human physical and mental power of humans with the latest developments of the life science , information science , electronic science , cognitive neuroscience and other fields .This paper provides an overview of status quo of human enhancement technology and its military applica -tions, and analyzes the factors that may adversely affect its applications .

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