1.Role of PI3K/Akt Pathway in Epirubicin Resistance in Triple-Negative Breast Cancer Explored Through Transcriptomic Analysis
Lingshan NAN ; Xiaomin WANG ; Xi ZUO ; Haiming LI ; Dong CHEN ; Xiaohui YIN ; Ganlin ZHANG
Cancer Research on Prevention and Treatment 2026;53(5):339-348
Objective To establish an epirubicin (EPI)-resistant murine triple-negative breast cancer (TNBC) (4T1/EPI) cell line and evaluate its biological characteristics and drug resistance. Methods The EPI-resistant cell line 4T1/EPI was developed through intermittent induction with gradually increasing EPI concentrations in vitro. Morphological changes were observed under an inverted microscope. Drug resistance index (MTT assay), cell doubling time (CCK-8 assay), and migration ability (wound healing assay) were evaluated. Western blot was used to detect the expression of drug resistance-related proteins. Transcriptome sequencing and KEGG pathway enrichment analysis were performed to identify the pathways and targets involved in EPI resistance, followed by experimental validation. Results The 4T1 cells eventually grew normally in a medium containing 100 ng/mL EPI, confirming the establishment of the 4T1/EPI resistant cell line. After stable resistance was acquired, morphological alterations were observed. Compared with their parental 4T1 cells, 4T1/EPI cells showed significantly prolonged doubling time (P<0.01) and enhanced migration ability (P<0.05). Expression levels of drug resistance-related proteins MDR1, MRP1 (P<0.01), and ABCG2 (P<0.05) were elevated in 4T1/EPI cells. In vivo models also demonstrated significant EPI resistance in 4T1/EPI tumors in terms of tumor weight and volume. Transcriptome sequencing highlighted the involvement of the PI3K/Akt signaling pathway and ABC transporter pathway. Validation experiments showed the upregulation of Erbb3, Egfr, PI3K, and Akt (P<0.05) and significant downregulation of Fgfr1 (P<0.01) in 4T1/EPI cells. Conclusion The EPI-resistant TNBC cell line 4T1/EPI was successfully established, exhibiting significant resistance in vitro and in vivo. The mechanism may involve the EPI-induced upregulation of Egfr and Erbb3, activating the PI3K/Akt pathway and subsequently enhancing ABC transporter expression.
2.Clinical efficacy of using removable titanium clip combined with rubber band and endoscopic double traction for treating gastric submucosal tumor
Xiaoling CHEN ; Xiaoyan FU ; Lingshan HUANG ; Jing YU
China Journal of Endoscopy 2025;31(5):66-71
Objective To study the clinical efficacy and safety of detachable titanium clip combined with rubber band intracavity dual traction assistance in endoscopic treatment of gastric submucosal tumors(SMT).Methods 52 patients who underwent endoscopic submucosal excavation(ESE)or endoscopic full thickness resection(EFR)for gastric SMT from January 2022 to July 2024 were selected.27 patients using the detachable titanium clip combined with rubber band intracavity dual traction assistance ESE or EFR were selected as experimental group,and 25 patients using the traditional ESE or EFR were selected as control group.Operation time,hospital stay,hospital cost,one time complete excavation rate and complication rates were compared between the two groups.Results The operation time and hospital stay in the experimental group were significantly shorter than those in the control group,and the hospital cost was also significantly lower,with all differences being statistically significant(P<0.05).The comparison of surgical complication rates between the two groups showed a statistically significant difference(P<0.05),with the experimental group having significantly lower total complication rates than that in the control group.However,there was no statistically significant difference(P>0.05)in the rate of one-time complete excavation between the two groups.Conclusion The detachable titanium clip combined with rubber band intracavitary dual-traction-assisted endoscopic treatment for gastric SMT significantly shortens the operation time and hospital stay,reduces hospital costs and surgical complications.It is a simple,effective auxiliary device that worthy for clinical application.
3.Preparation of monoclonal antibody against PRRSV-2 N protein and identification of antigenic epitopes
Yanli PANG ; Jianguang QIN ; Muyang LIU ; Tongwei REN ; Jiaqi LIU ; Lingshan ZHOU ; Ying CHEN ; Kang OUYANG ; Weijian HUANG ; Zuzhang WEI
Chinese Journal of Veterinary Science 2025;45(1):16-21,45
To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.
4.Isolation,identification and biological characteristics of Aeromonas hydrophila from South China tiger
Yuqi LI ; Yali KANG ; Yubin ZHUO ; Lingshan HUANG ; Shuqi QIU ; Yuxi XUE ; Xiaop-ing WU ; Sisi FAN ; Yuting LIAO ; Weiye LIN ; Chan CHEN ; Kaixiong LIN ; Tengteng CHEN ; Xipan LIN ; Kewei FAN
Chinese Journal of Veterinary Science 2025;45(1):46-52,58
The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2%to 99.1%,with the highest homology of 99.1%observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.
5.Clinical efficacy of using removable titanium clip combined with rubber band and endoscopic double traction for treating gastric submucosal tumor
Xiaoling CHEN ; Xiaoyan FU ; Lingshan HUANG ; Jing YU
China Journal of Endoscopy 2025;31(5):66-71
Objective To study the clinical efficacy and safety of detachable titanium clip combined with rubber band intracavity dual traction assistance in endoscopic treatment of gastric submucosal tumors(SMT).Methods 52 patients who underwent endoscopic submucosal excavation(ESE)or endoscopic full thickness resection(EFR)for gastric SMT from January 2022 to July 2024 were selected.27 patients using the detachable titanium clip combined with rubber band intracavity dual traction assistance ESE or EFR were selected as experimental group,and 25 patients using the traditional ESE or EFR were selected as control group.Operation time,hospital stay,hospital cost,one time complete excavation rate and complication rates were compared between the two groups.Results The operation time and hospital stay in the experimental group were significantly shorter than those in the control group,and the hospital cost was also significantly lower,with all differences being statistically significant(P<0.05).The comparison of surgical complication rates between the two groups showed a statistically significant difference(P<0.05),with the experimental group having significantly lower total complication rates than that in the control group.However,there was no statistically significant difference(P>0.05)in the rate of one-time complete excavation between the two groups.Conclusion The detachable titanium clip combined with rubber band intracavitary dual-traction-assisted endoscopic treatment for gastric SMT significantly shortens the operation time and hospital stay,reduces hospital costs and surgical complications.It is a simple,effective auxiliary device that worthy for clinical application.
6.Preparation of monoclonal antibody against PRRSV-2 N protein and identification of antigenic epitopes
Yanli PANG ; Jianguang QIN ; Muyang LIU ; Tongwei REN ; Jiaqi LIU ; Lingshan ZHOU ; Ying CHEN ; Kang OUYANG ; Weijian HUANG ; Zuzhang WEI
Chinese Journal of Veterinary Science 2025;45(1):16-21,45
To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.
7.Isolation,identification and biological characteristics of Aeromonas hydrophila from South China tiger
Yuqi LI ; Yali KANG ; Yubin ZHUO ; Lingshan HUANG ; Shuqi QIU ; Yuxi XUE ; Xiaop-ing WU ; Sisi FAN ; Yuting LIAO ; Weiye LIN ; Chan CHEN ; Kaixiong LIN ; Tengteng CHEN ; Xipan LIN ; Kewei FAN
Chinese Journal of Veterinary Science 2025;45(1):46-52,58
The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2%to 99.1%,with the highest homology of 99.1%observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.
8.A qualitative study on the experience and needs of dignity-conserving care for the disabled elderly people in nursing homes
Lingshan CHEN ; Cuizhen SHEN ; Qianyin ZHU ; Tingyu MU
Chinese Journal of Nursing 2023;58(24):3004-3011
Objective This study aimed to explore the experience and needs of dignity-conserving care of disabled elderly people in nursing homes,and to provide a basis for formulating a quality evaluation system for dignity-conserving care.Methods This descriptive qualitative study,involving a semi-structured in-depth interview of 13 disabled elderly people based on purposive sampling,was conducted from August to November 2022 at a nursing home in Hangzhou.Based on the harmony nursing theory,the directional content analysis method was performed in order to summarize and extract themes from the interview texts.Results The dignity-conserving care experience and needs of disabled elderly people were extracted and classified into 5 themes,including physical safety(physical health maintenance,privacy protection,hygiene and image maintenance),psychological health(psychological counseling,psychological support),social communication(well communication environment,harmonious way of communication),spiritual culture(self-realization,respect for faith),needs of dignity-conserving care(individualized health needs,activity participation needs,dignified communication needs,spiritual needs of respecting the elderly).Conclusion When conducting the dignity-conserving care,nursing homes should pay attention to the physical health of the disabled elderly,actively maintain the image and privacy of the disabled elderly people,pay attention to the needs of psychological health,provide psychological counseling and support,strengthen the construction of barrier-free environments and cultural environments,build mutual respect way of communication,fully respect the religious beliefs of disabled elderly people,promote the realization of self-value of disabled elderly people,broaden the channels of personalized service and activity participation,strengthen the interaction of the dignity and spirit of the respect for the elderly of nursing homes,comprehensively improve the quality of dignity-conserving care in nursing homes.
9.Carotid stiffening predicts cardiovascular risk stratification in mid-life: non-invasive quantification with ultrafast ultrasound imaging
Zhengqiu ZHU ; Lingshan CHEN ; Wenjun LIU ; Yiyun WU ; Chong ZOU ; Xinyi ZHANG ; Shanshan HE ; Yinping WANG ; Bixiao SHEN ; Xuehui MA ; Hui GAO ; Yun LUAN ; Hui HUANG
Ultrasonography 2022;41(3):462-472
Purpose:
The present study investigated the association between Systematic COronary Risk Evaluation (SCORE)-estimated cardiovascular risk and carotid stiffening in a middle-aged population using ultrafast pulse wave velocity (ufPWV).
Methods:
This study enrolled 683 participants without known cardiovascular disease or diabetes mellitus who underwent ufPWV measurements. Clinical interviews, physical examinations, laboratory findings, carotid intima-media thickness (cIMT), pulse wave velocity (PWV) at the beginning of systole (PWV-BS), and PWV at the end of systole (PWV-ES) were assessed. Each participant underwent an assessment of SCORE risk based on major cardiovascular risk factors (CVRFs), including age, sex, smoking, systolic blood pressure (SBP), and total cholesterol (TC). Crude and adjusted odds ratios (ORs) with 95% confidence intervals and ordinal logistic regression were used. Overall CVRFs were adjusted to assess ORs.
Results:
cIMT and carotid stiffening in PWV-BS and PWV-ES were significantly different between sex subgroups (all P<0.05), but only PWV-ES increased gradually in age and SCORE-estimated risk subgroups (all P<0.05). Compared with cIMT (r=0.388, P<0.001) and PWV-BS (r=0.159, P<0.001), PWV-ES was more strongly correlated with SCORE categories (r=0.405, P<0.001). Higher PWV-ES values were associated with SCORE categories independently of sex, SBP, TC, and smoking in moderate-risk and high-risk subgroups (OR, 1.63; P<0.001 and OR, 2.12; P=0.024, respectively), but were not independent of age in all risk subgroups (all P>0.05).
Conclusion
Carotid stiffening quantified by ufPWV is linked to SCORE categories, and elevated PWV-ES may aid in cardiovascular risk stratification.
10.Effects of Human Immunodeficiency Virus-positive Mothers Receiving Antiretroviral Therapy to Prevent Mother-to-child Transmission on the Growth and Development of 18-month-old Children in Lingshan County of Guangxi.
Xian-Min GE ; Mei LIN ; Jin-Mei CHEN ; Jin-Hui ZHU ; Guang-Hua LAN ; Yu YAN ; Huan-Huan CHEN ; Liu-Hong LUO ; Shan-Shan LI ; Bin LI ; Guang-Hua HUANG ; Jia-Wei LI ; Yue-Qin DENG ; Ping-Jing WEN ; Yu-Man HUANG ; Qiu-Ying ZHU ; Qin MENG ; Shuai-Feng LIU ; Xiu-Ling WU ; Jing-Hua HUANG ; Hua-Xiang LU
Acta Academiae Medicinae Sinicae 2021;43(5):743-748
Objective To evaluate the effects of antiretroviral therapy(ART)for the prevention of mother-to-child transmission(PMTCT)of acquired immune deficiency syndrome(AIDS)on the growth and development of 18-month-old children born by human immunodeficiency virus(HIV)-positive pregnant women in Lingshan County,Guangxi Zhuang Autonomous Region,and provide scientific evidence for improving the ART medication plan for PMTCT.Methods Lingshan County,ranking the first in the HIV-epidemic counties of Guangxi,was selected as the research site.According to the design of retrospective case-control study,we assigned all the subjects into the case group and the control group:(1)The case group included the HIV-positive pregnant women who had received ART for PMTCT and their HIV-negative infants in Lingshan County from 2010 to 2017.The historical cards and PMTCT data of them were collected from the national PMTCT database.(2)The control group included the healthy pregnant women and their healthy babies born in the Lingshan Maternity and Infant Hospital in 2017,and the children's growth and development data were collected.The stunted growth in children was defined as at least one of the three main indicators of body height,body weight,and head circumference below the normal range.Results The number of HIV-positive mothers and their infants in the case group was 391 and 368,respectively,and 87.21%(341/391)and 95.38%(351/368)of mothers and infants respectively received ART medication.The HIV positive rate,mortality rate,and mother-to-child transmission rate of 18-month-old children were 1.36%(5/368),4.35%(16/368),and 2.01%(5/249),respectively.The incidence of stunted growth of 18-month-old children in the case group and the control group was 42.12%(155/368)and 23.06%(101/438),respectively,with significant difference(
Case-Control Studies
;
China/epidemiology*
;
Female
;
Growth and Development
;
HIV
;
HIV Infections/prevention & control*
;
Humans
;
Infant
;
Infectious Disease Transmission, Vertical/prevention & control*
;
Mothers
;
Pregnancy
;
Pregnancy Complications, Infectious/drug therapy*
;
Retrospective Studies

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