1.Explore the mechanism of Huangqi Jiedu decoction in the treatment of breast cancer syndrome of Zheng-deficiency and toxic-incandescence based on network pharmacology and molecular docking technology
Lingjia TIAN ; Zihao YIN ; Liang ZHOU ; Qin ZHANG ; Lixin LIANG ; Yongjie CAO ; Hua WANG ; Lifang LIU
Journal of Chinese Physician 2024;26(3):360-365
Objective:To explore the mechanism of Huangqi Jiedu Decoction (HQJD) in the treatment of breast cancer with the syndrome of Zheng deficiency and toxic incandescence by network pharmacology and molecular docking technology.Methods:The main active ingredients and targets of HQJD were screened through the traditional Chinese medicine (TCM) systematic pharmacology database and analysis platform. The relevant targets of breast cancer with the syndrome of Zheng-deficiency, toxic-incandescence were obtained using OMIM, GeneGards and Drugbank databases, and the relevant targets of HQJD for the treatment of breast cancer with the syndrome of Zheng-deficiency and toxic incandescence were obtained by intersection; The Cytoscape 3.9.1 software was used to build the protein protein interaction (PPI) network and the " drug active component target disease" network on the basis of String 11.0 database, and the core active components and core targets of HQJD in treating breast cancer with the syndrome of Zheng-deficiency and toxic-incandescence were inferred according to the topological parameters. gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on core targets using R language; and molecular docking verification on the main active ingredients and core targets were conducted.Results:230 effective targets of active ingredients of HQJD were screened, and 15 467 active ingredients of breast cancer with syndrome of Zheng-deficiency/toxic-incandescence were obtained; 217 intersection targets; GO function enrichment analysis showed that the treatment of HQJD for breast cancer with the syndrome of Zheng-deficiency and toxic-incandescence mainly involved oxidative stress and cytochemical stress; The enrichment analysis of KEGG pathway showed that HQJD treatment of breast cancer with the syndrome of Zheng-deficiency and toxic-incandescence was mainly related to phosphatidylinositol 3-protein kinase B (PI3K-Akt), interleukin-17 (IL-17) and other signal pathways. The molecular docking results showed that the main active ingredients such as β-sitosterol, stigmasterol, luteolin had good binding ability with core targets.Conclusions:HQJD has the characteristics of multi-component, multi target and multi pathway in the treatment of breast cancer with syndrome of Zheng-deficiency and toxic-incandescence, and its main mechanism may be related to PI3K-Akt, IL-17, P53 and other signal pathways.
2.Preliminary application of pseudovirus in evaluating the immunogenicity of recombinant poliovirus vaccines and in rat potency tests
Yueyue LIU ; Lifang DU ; Shi CHEN ; Rongrong ZHAO ; Fan ZHENG ; Ling DING ; Zhijing MA ; Jing ZHANG ; Meng MA ; Suhua ZHAO ; Yu LIANG ; Qiming LI ; Yaru QUAN
Chinese Journal of Microbiology and Immunology 2024;44(9):778-783
Objective:To preliminarily investigate the applicability of a poliovirus pseudovirus-based neutralization assay in evaluating the immunogenicity of recombinant poliovirus vaccines and their in vivo potency in rats. Methods:Serum samples from rats immunized with recombinant poliovirus vaccines were tested using both the pseudovirus neutralization assay and the live-virus neutralization assay with Sabin strain. The consistency and correlation of the two methods were analyzed using the Kappa test and Spearman′s rank correlation.Results:For the neutralizing antibodies against typeⅠ, Ⅱ, and Ⅲ polioviruses, the Kappa values for consistency analysis of the two methods were 0.914, 1.000, and 0.751, respectively ( P<0.001), and the correlation coefficients ( R values) were 0.833, 0.927, and 0.859, respectively ( P<0.001). Conclusions:The test results of the two methods are consistent and show a good correlation, indicating that the pseudovirus neutralization assay can be applied to evaluating the immunogenicity of poliovirus vaccines and also can be used in rat potency tests.
3.Clinical application of EGFR co-mutation gene analysis in lung cancer patients based on circulating tumor DNA
Dan LI ; Liang SHAN ; Lifang MA ; Jiayi WANG
Chinese Journal of Laboratory Medicine 2024;47(11):1286-1291
Objective:To investigate the clinical significance of gene mutations in epidermal growth factor receptor (EGFR), tumor suppressor gene P53 (TP53), phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), or SMAD family member 4 (SMAD4) in patients with advanced non-small cell lung cancer (NSCLC) based on circulating tumor DNA (ctDNA) analysis.Methods:A single-center, retrospective study was conducted among patients with NSCLC who visited Shanghai Chest Hospital from June 2022 to January 2024. Peripheral blood was collected from patients prior to targeted therapy, and cell-free DNA (cfDNA) and leukocyte DNA were isolated for next-generation sequencing. ctDNA data was obtained through comparative analysis. Patient gene mutations were analyzed according to the guidelines for the interpretation of somatic variants jointly established by the Association for Molecular Pathology/College of American Pathologists/American Society of Clinical Oncology. Analysis of variance and Kaplan-Meier survival curves were applied to assess the partial response (PR) rate among patients with EGFR gene mutations and those with concurrent mutations in other genes.Results:Among 126 patients, the rate of carrying only EGFR mutations was 46.0% (58/126), with a PR rate of 74.1% (43/58). The rate of double mutations in EGFR and TP53 was 37.3% (47/126), with a PR rate of 53.2% (25/47). The rate of triple mutations in EGFR, TP53, and concurrent mutations in PIK3CA or SMAD4 was 16.7% (21/126), with a PR rate of 28.6% (6/21). The comparison of progression-free survival curves among the three groups showed a statistically significant difference (χ 2=23.81, P<0.01). Conclusions:next-generation sequencing detection technology based on ctDNA can detect concurrent mutations in TP53, PIK3CA, or SMAD4 along with EGFR mutations. Multiple gene mutations are significant factors for poor PR rates and progression-free survival curves in patients.
4.Immunogenicity and immune persistence of purified Vero cell rabies vaccine with Zagreb and Essen regimens
Jingxia LUO ; Xinjun LYU ; Lifang ZHANG ; Weixian LIANG ; Xuemei ZENG ; Haijin LU ; Liling WEI ; Guirong LIANG ; Qinglian LEI
Chinese Journal of Experimental and Clinical Virology 2021;35(6):612-618
Objective:To evaluate the immunogenicity and immune persistence of purified Vero cells rabies vaccine (PVRV) with Zagreb and Essen regimen.Methods:Prospective study: Patients with first Class II exposure to rabies were recruited from the Dog Injury Cinic of Guangxi Center for Disease Control and Prevention (Guangxi CDC) and randomly divided into the Zagreb (2-1-1) and Essen (1-1-1-1-1) regimen group. All patients were inoculated with the vaccines from the same manufacturer and batch, and 3 ml serum was collected at the 45th day and in 1-year, 2-year and 3-year after immunization. Rapid fluorescent inhibition test (RFFIT) was used to detect rabies virus neutralizing antibody (RVNA). The attenuation of RVNA positive rate and geometric mean titer (GMT) with time was analyzed. Retrospective study: The informed consents for rabies vaccine in the Dog Injury Clinic of Guangxi CDC were checked out. The patients who were injected with PVRV (same manufacturer but unlimited batch) but without passive immune agents for the first time within 3 years were selected and divided into 1-year, 2-year and 3-year group. Each group was further divided into Zagreb and Essen regimen group. The serum (3 ml) was collected at 1 year, 2 years and 3 years after immunization and detected the RVNA by RFFIT.Results:Prospective study: The RVNA positive rates on the 45th day and in 1-year, 2-year and 3-year after immunizationin in the Zagreb and Essen regimen group were 100%, 95%, 85%, 80% and 98.25%, 89.47%, 89.47%, 85.96%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT on the 45th day and in 1-year, 2-year, and 3-year in the Zagreb and Essen regimen group were 11.32 IU/ml, 1.69 IU/ml, 1.30 IU/ml, 1.30 IU/m and 13.18 IU/ml, 2.13 IU/ml, 1.87 IU/ml, 1.84 IU/m, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( F=1.971, P=0.164). The RVNA GMT levels in the two regimen groups had the same trend of attenuation over time (time*group F=0.702, P=0.435). Retrospective study: The RVNA positive rates in 1-year, 2-year and 3-year after immunization in the Zagreb and Essen regimen group were 100%, 95%, 91.43%和94.73%, 86.21%, 87.5%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT in 1-year, 2-year, and 3-year groups after immunization in the Zagreb and Essen regimen group were 2.65 IU/ml, 2.03 IU/ml, 1.57 IU/ml和3.2 IU/ml, 2.58 IU/ml, 2.45 IU/ml, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( P>0.05). Conclusions:The PVRV showed the same excellent immunogenicity and immune persistence after the vaccination with the Zagreb and Essen regimens.
5.Application of scenario simulation teaching in cardiopulmonary resuscitation training for junior surgical residents
Suhua KUANG ; Shaobo XIE ; Lifang WENG ; Xiaoshan REN ; Rong LIU ; Qing LIANG
Chinese Journal of Medical Education Research 2019;18(6):636-640
Objective To explore the effect of scenario simulation teaching method in the training of cardiopulmonary resuscitation (CPR) for junior surgical residents. Methods 133 junior residents (working life<3 years) in the author's hospital rotating Cardiac Surgery department were selected and divided into control group (n=65) and observation group (n=68), in which the control group adopted traditional classroom teaching: teacher explanation-demonstration-student practice-teacher counseling; the observation group adopted scenario simulation teaching: teacher explanation-demonstration-student practice-teacher counseling scenario simulation. Before and after training, two groups both received the CPR theory and double operation assessment , comparing the difference of assessment scores between groups before and after training . Results There was no statistically significant difference in the demographic characteristics between the two groups. The assessment scores of CPR theory and operation were improved in both groups after training, and there were significant differences (P=0.000). There were no statistically significant differences in the CPR theory and operation assessment scores between the two groups before training (P>0.05);compared with the scores of CPR operation between the two groups after training, the observation group [(84.62±3.94)] was significantly higher than that of the control group [(79.68±5.45)] and there were significant statistical differences (P=0.000), while there was no statistically significant difference in CPR theory assessment scores between the two groups (P>0.05). Conclusions Both traditional classroom teaching and scenario simulation teaching methods could improve the CPR theory and skill level of the surgical junior resident, but situa tional simulation teaching method is better than traditional classroom teaching method in improving students' CPR performance.
6.The in vivo thrombosis evaluation for the biodegradable polymer stent.
Liping ZHENG ; Lifang JIA ; Tun YUAN ; Jie LIANG
Journal of Biomedical Engineering 2019;36(2):232-237
New biodegradable intravascular stent can reduce risk of foreign bodies retained, thus, it is widely concerned and some of the products have been introduced into the clinic. However, the characteristic of biodegradable may lead to more safety concerns associated with thrombosis. To ensure the safety, the thrombus formation experiment needs to be carefully designed and evaluated based on GB/T 16886.4 standard, but current standard do not provide explicit testing and evaluating methods. Establishing animal model with experimental pigs, the study compares biodegradable coronary stents and metal stents by simulating clinical implantation on the thrombus formation in the implanting process, and after the short-term and long-term implantation. The evaluation methods include gross observation, digital subtraction angiography intraoperative analysis, optical coherence tomography analysis, scanning electron microscopy and so on. The results show that combining these methods could comprehensively evaluate the whole process of the thrombus formation from the beginning of implantation to the end of preclinical animal experiments, so that, it may better predict the clinical thrombosis risk, and the selection of the control was very important. The study tries to use the comparison examples of thrombosis on the new medical instrument to provide the clue for thrombosis evaluation on similar instruments and show the methodology on the preclinical evaluation.
Absorbable Implants
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adverse effects
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Animals
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Drug-Eluting Stents
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adverse effects
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Polymers
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Swine
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Thrombosis
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etiology
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Tomography, Optical Coherence
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Treatment Outcome
7.Correlation analysis and clinical significance of lymph node metastasis in right recurrent laryngeal nerve of papillary thyroid carcinoma
Hongcun CHEN ; Liang LI ; Ming JIANG ; Jun ZHANG ; Baozhong YAO ; You JIANG ; Lifang LIAO
Journal of International Oncology 2018;45(7):391-394
Objective To explore the correlation factors and clinical significance of lymph node metastasis in right recurrent laryngeal nerve of thyroid papillary carcinoma (PTC).Methods Ninty-eight consecutive patients with PTC who were underwent total thyroidectomy with routine central lymph node dissection in the Second People's Hospital of Hefei from January 2015 to August 2017 were analyzed.The right paratracheal lymph nodes in the central compartment lymph nodes were divided into the level Ⅵ-A (anterior) and level Ⅵ-B (posterior,that was lymph node posterior to recurrent laryngeal nerve) compartments by recurrent laryngeal nerve.The lymph node metastasis of Ⅵ-B area during central compartment lymph node dissection was analyzed.We drew the receiver-operating characteristic curve (ROC) for right neck Ⅵ-A number of lymph node metastasis,and calculated the area under the curve (AUC) and Youden index.Results Among 98 cases,16 cases occurred Ⅵ-B district lymph node metastasis (16.33%).Single factor analysis results showed that lymph node metastasis in Ⅵ-B area of PTC patients were related to the tumor size (x2 =12.864,P <0.001),tumor capsular invasion (x2 =16.354,P < 0.001),the right neck Ⅵ-A area lymph node metastasis (x2 =16.065,P < 0.001),tumor number (x2 =15.593,P < 0.001) and neck lymph node metastasis (x2 =21.098,P <0.001),but they were not related to the patients' gender,age and lesion location (all P > 0.05).Lymph node metastasis in Ⅵ-B area of PTC patients were related to the number of right neck Ⅵ-A area lymph node metastasis.When the number of right neck Ⅵ-A metastatic lymph nodes was 2.5,the sensitivity and specificity were 70.60% and 70.00% respectively,AUC was 0.754,and Youden index was 0.406.Conclusion For patients with PTC,primary tumor diameter > 1 cm,tumor extracapsular invasion,Ⅵ-A area lymph node metastasis,multiple tumor and lateral cervical lymph node metastasis were the predictive factors for the lymph node metastasis in Ⅵ-B area.When the number of right neck Ⅵ-A area metastatic lymph nodes was greater than 3,we should dissect Ⅵ-B area.
8.Application of miRNA-34a, miRNA-34c and miRNA-135a in early diagnosis of mild cognitive impairment and Alzheimer's disease
Lifang CHEN ; Lu ZHANG ; Manfu HAN ; Junming YIN ; Shiyu HU ; Zhuoyuan LIANG
Journal of Chinese Physician 2017;19(5):683-686
Objective To investigate the clinical value of miRNA-34a,miRNA-34c,and miRNA-135a in the early diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods The patients were collected in the outpatient and inpatient of Neurology during Aug 2014 to May 2016.The levels of miRNA-34a,miRNA-34c,an dmiRNA-135a were detected with quantitative real-time polymerase chain reaction (qRT-PCR).Results AD patients had higher level of miRNA-34a than the controls.The levels of miRNA-34c were higher in MCI and AD patients,while lower levels of miRNA-135a compared to the controls.Conclusions The miRNA 34a and miRNA-135a were likely to became the biomarker in early diagnosis of MCI and AD.
9.Expression and clinical significance of MIP-1 alpha in serum of asthmatic patients
Xinhuai XIAO ; Xiaomin LIANG ; Lifang SU
Journal of Chinese Physician 2017;19(8):1200-1202,1208
Objective To investigate the levels of serum macrophage inflammatory protein-1α (MIP-1α),inflammatory factors and pulnonary function in patients with asthma,and the correlation between thenm.Methods Eighty patients with asthma were selected who were treated in our hospital from March 2015 to August 2016,and the data of 80 healthy adults were selected as control.The differences of serum MIP-1α,inflammatory factors,imnmunoglobulin and lung function were observed in two groups.The correlation between MIP-1α and inflammatory factors,lung function and immunoglobulin were analyzed.Results The levels of MIP-1α,interleukin (IL)-6,and tumor necrosis factor-α (TNF-α) in patients with asthma were significantly higher than those in control group (t =-207.04,-33.209,-55.132,P < 0.01);The levels of forced vital capacity (FVC),the forced expiratory volume in one second (FEV1),FEV1/FVC,maximal mid expiratory flow (MMEF),and peak expiratory flow (PEF) in the asthma group were significantly lower than those in the control group (t =17.100,39.154,22.791,25.391,19.356,P < 0.01).The IgG and IgM of asthma group were significantly lower than those of the control group (t =9.564,7.528,P < 0.01).The IgE level was significantly higher than that of the control group (t =-82.683,P <0.01).The level of MIP-1α was positively correlated with the levels of IL-6,TNF-0 and IgE,and negatively correlated with FVC,FEV1,FEV1/FVC,MMEF,PEF,IgG and IgM in patients with asthma.Conclusions The level of serum MIP-1 α in asthmatic patients is high,and is closely related to the inflammatory cytokines and lung function in patients with asthma.
10.Constructing a recombinant adenovirus carrying human cytomegalovirus UL138 gene and evaluating the effects of UL138 gene on the functions of monocytes
Gangqiang GUO ; Jing CHEN ; Wenjing CHEN ; Lulu YE ; Xiangwei SUN ; Liang ZHANG ; Huidi ZHANG ; Qiao′ai LIN ; Xian SHEN ; Lifang ZHANG ; Xiangyang XUE
Chinese Journal of Microbiology and Immunology 2016;36(9):667-675
Objective To construct a recombinant adenovirus carrying UL138 gene, which was re-lated to the latent infection of human cytomegalovirus, and to investigate the effects of UL138 gene on the functions of THP-1 mononuclear cells. Methods The recombinant adenovirus expressing the UL138 gene was packaged. The titer of the recombinant adenovirus was determined by calculating 50% tissue culture in-fective dose ( TCID50 ) . THP-1 mononuclear cells were infected with the recombinant adenovirus at different multiplicity of infection (MOI) and the optimal MOI was determined (100 PFU/cell) by observing the ex-pression of green fluorescent protein ( GFP ) . Changes in the expression of proinflammatory cytokines by THP-1 mononuclear cells that was induced by overexpressed UL138 were analyzed by quantitative PCR. The expression of chemokines and their receptors were measured by quantitative PCR array. Results The re-combinant adenovirus carrying the UL138 gene was successfully constructed with a titer of 1×1011 PFU/ml. The rate of THP-1 mononuclear cells that was infected with the recombinant adenovirus was 60% at the MOI of 1 ∶ 100. Results of the RT-PCR analysis and Western blot assay further confirmed that the recombinant adenovirus could infect THP-1 mononuclear cells successfully and the expression of UL138 protein increased gradually over time. The overexpressed UL138 in THP-1 mononuclear cells significantly inhibited the expres-sion of IL-18, IL-1β, IL-6, IL-8 and TNF-α as indicated by the results of quantitative PCR. Results ob-tained from the quantitative PCR array analysis showed that most of the chemokines and their receptors were downregulated in the transfected THP-1 mononuclear cells except for the chemokines of CCL17, CCL21, CCL2 and XCL2 and the receptors of CCR2, CXCR1, CXCR2, CXCR4 and CX3CR1 which were upregulat-ed. Conclusion We successfully constructed the recombinant adenovirus carrying UL138 gene which could be used to infect THP-1 mononuclear cells. Overexpressed UL138 in THP-1 mononuclear cells significantly affected the functions of THP-1 mononuclear cells.

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