1.Gene Expression Alteration by Non-thermal Plasma-Activated Media Treatment in Radioresistant Head and Neck Squamous Cell Carcinoma
Sicong ZHENG ; Yudan PIAO ; Seung-Nam JUNG ; Chan OH ; Mi Ae LIM ; QuocKhanh NGUYEN ; Shan SHEN ; Se-Hee PARK ; Shengzhe CUI ; Shuyu PIAO ; Young Il KIM ; Ji Won KIM ; Ho-Ryun WON ; Jae Won CHANG ; Yujuan SHAN ; Lihua LIU ; Bon Seok KOO
Clinical and Experimental Otorhinolaryngology 2025;18(1):73-87
Objectives:
. Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes.
Methods:
. After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC.
Results:
. NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways.
Conclusion
. NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.
2.Gene Expression Alteration by Non-thermal Plasma-Activated Media Treatment in Radioresistant Head and Neck Squamous Cell Carcinoma
Sicong ZHENG ; Yudan PIAO ; Seung-Nam JUNG ; Chan OH ; Mi Ae LIM ; QuocKhanh NGUYEN ; Shan SHEN ; Se-Hee PARK ; Shengzhe CUI ; Shuyu PIAO ; Young Il KIM ; Ji Won KIM ; Ho-Ryun WON ; Jae Won CHANG ; Yujuan SHAN ; Lihua LIU ; Bon Seok KOO
Clinical and Experimental Otorhinolaryngology 2025;18(1):73-87
Objectives:
. Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes.
Methods:
. After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC.
Results:
. NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways.
Conclusion
. NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.
3.Gene Expression Alteration by Non-thermal Plasma-Activated Media Treatment in Radioresistant Head and Neck Squamous Cell Carcinoma
Sicong ZHENG ; Yudan PIAO ; Seung-Nam JUNG ; Chan OH ; Mi Ae LIM ; QuocKhanh NGUYEN ; Shan SHEN ; Se-Hee PARK ; Shengzhe CUI ; Shuyu PIAO ; Young Il KIM ; Ji Won KIM ; Ho-Ryun WON ; Jae Won CHANG ; Yujuan SHAN ; Lihua LIU ; Bon Seok KOO
Clinical and Experimental Otorhinolaryngology 2025;18(1):73-87
Objectives:
. Head and neck squamous cell carcinoma (HNSCC) exhibits high recurrence rates, particularly in cases of radioresistant HNSCC (RR-HNSCC). Non-thermal plasma (NTP) therapy effectively suppresses the progression of HNSCC. However, the therapeutic mechanisms of NTP therapy in treating RR-HNSCC are not well understood. In this study, we explored the regulatory role of NTP in the RR-HNSCC signaling pathway and identified its signature genes.
Methods:
. After constructing two RR-HNSCC cell lines, we prepared cell lysates from cells treated or not treated with NTP-activated media (NTPAM) and performed RNA sequencing to determine their mRNA expression profiles. Based on the RNA sequencing results, we identified differentially expressed genes (DEGs), followed by a bioinformatics analysis to identify candidate molecules potentially associated with NTPAM therapy for RR-HNSCC.
Results:
. NTPAM reduced RR-HNSCC cell viability in vitro. RNA sequencing results indicated that NTPAM treatment activated the reactive oxygen species (ROS) pathway and induced ferroptosis in RR-HNSCC cell lines. Among the 1,924 genes correlated with radiation treatment, eight showed statistical significance in both the cell lines and The Cancer Genome Atlas (TCGA) cohort. Only five genes—ABCC3, DUSP16, PDGFB, RAF1, and THBS1—showed consistent results between the NTPAM data sequencing and TCGA data. LASSO regression analysis revealed that five genes were associated with cancer prognosis, with a hazard ratio of 2.26. In RR-HNSCC cells, NTPAM affected DUSP16, PDGFB, and THBS1 as activated markers within 6 hours, and this effect persisted for 12 hours. Furthermore, enrichment analysis indicated that these three DEGs were associated with the extracellular matrix, transforming growth factor-beta, phosphoinositide 3-kinase/protein kinase B, and mesenchymal-epithelial transition factor pathways.
Conclusion
. NTPAM therapy exerts cytotoxic effects in RR-HNSCC cell lines by inducing specific ROS-mediated ferroptosis. DUSP16, PDGFB, and THBS1 were identified as crucial targets for reversing the radiation resistance induced by NTPAM therapy, providing insights into the mechanisms and clinical applications of NTPAM treatment in RR-HNSCC.
4.Prevalence and genotyping of Cryptosporidium spp. and Giardia lamblia in dogs and cats from a pet hospital in Shanghai Municipality
Jing ZHANG ; Yuan QIN ; Yujuan SHEN ; Yaxue WANG ; Jianping CAO ; Yaxin SU ; Hua LIU
Chinese Journal of Schistosomiasis Control 2023;35(3):258-262
Objective To investigate the prevalence and genotypes of Cryptosporidium spp. and Giardia lamblia in dogs and cats from a pet hospital in Shanghai Municipality. Methods A total of 145 fresh fecal samples were collected from pet dogs and cats in a pet hospital in Shanghai Municipality during the period from November 2021 to June 2022, including 99 dog fecal samples and 46 cat fecal samples. The small subunit ribosomal ribonucleic acid (SSU rRNA) gene of Cryptosporidium and the triose phosphate isomerase (TPI) gene of G. lamblia were amplified using nested PCR assay, and the positive amplification products were sequenced from both directions. The sequence assembly was performed using the software Clustal X 2.1, and sequence alignment was conducted using BLAST. A phylogenetic tree was created with the Neighbor-Joining method using MEGA 11.0 to identify parasite species or genotype. Results The overall prevalence of Cryptosporidium and G. lamblia was 20.00% (29/145) in 145 pet dog and cat fecal samples, with the prevalence of 0.69% (1/145) and 19.31% (28/145) in Cryptosporidium and G. lamblia, respectively. G. lamblia was only detected in dog fecal samples, with prevalence of 18.18% (18/99), while the detection rates of Cryptosporidium and G. lamblia were 2.17% (1/46) and 21.74% (10/46) in cat fecal samples. Nucleotide sequence analysis showed that one Cryptosporidium positive sample was characterized as C. felis, and 28 G. lamblia positive samples were all characterized as Giardia assemblage A, which showed 100% sequence homology with human isolates of Giardia. Phylogenetic analysis revealed that the sequences obtained in this study belonged to the same branch with the reported Giardia assemblage A. Conclusions Cryptosporidium and G. lamblia infection was prevalent in pet dogs and cats from the study pet hospital in Shanghai Municipality, and there is a zoonotic risk for the species and genotype. Intensified surveillance of Cryptosporidium and Giardia infection is recommended in pets and their owners, and improved management of pet keeping is required.
5.Retrospective analysis of multiple factors combination for early identification of pulmonary tuberculosis complicated with pulmonary embolism
Yujuan SHEN ; Jianfeng ZHONG ; Zhaowei TONG ; Fuchu QIAN ; Yong TONG ; Weihong WANG
Chinese Journal of Infectious Diseases 2022;40(6):361-365
Objective:To analyze the independent diagnostic indicators and their diagnostic values for pulmonary tuberculosis complicated with pulmonary embolism.Methods:A total of 34 cases of pulmonary tuberculosis complicated with pulmonary embolism treated in Huzhou Central Hospital from March 2014 to September 2021 were enrolled. And 136 patients with simple pulmonary tuberculosis who were hospitalized during the same period were collected with a ratio of 1∶4 according to the principle of age and gender matching. The general conditions, clinical symptoms, comorbidities and laboratory indicators of the patients were retrospectively analyzed. The univariate analysis was performed using independent samples t test, Mann-Whitney U test and chi-square test. Binary logistic regression was used to analyze the related diagnostic factors for pulmonary embolism in pulmonary tuberculosis patients, and the combined factors were constructed by transforming the model equation, and the receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value and evaluate its diagnostic value. Results:The univariate analysis showed that patients with pulmonary tuberculosis complicated with pulmonary embolism had higher ratio of chest tightness (67.6%(23/34) vs 22.1%(30/136)), syncope (23.5%(8/34) vs 0.7%(1/136)), fever (55.9%(19/34) vs 36.0%(49/136)), hemostatic drug use (100.0%(34/34) vs 13.2%(18/136)), history of venous thrombosis (8.8%(3/34) vs 0.7%(1/136)), atrial fibrillation (11.8%(4/34) vs 2.2%(3/136)) and D-dimer levels (4.090 0(1.035 0, 10.790 0) mg/L vs 0.850 0(0.432 5, 2.145 0) mg/L) than those of simple pulmonary tuberculosis patients, and the differences were all statistically significant ( χ2=26.35, 28.19, 4.47, 96.44, 7.75, 6.30 and Z=-4.65, respectively; all P<0.050). The arterial partial pressure of oxygen (PaO 2)(61.90(52.95, 73.00) mmHg vs 82.00 (75.00, 87.00) mmHg, 1 mmHg=0.133 kPa) and albumin ((28.83±4.98) g/L vs (32.76±5.65) g/L) of patients with pulmonary tuberculosis complicated with pulmonary embolism were lower than those of simple pulmonary tuberculosis patients, and the differences were both statistically significant ( Z=-5.21 and t=3.71, respectively, both P<0.001). Binary regression analysis showed that chest tightness (odds ratio ( OR)=3.494, 95%confidence interval ( CI) 1.208 to 10.100, P=0.021), D-dimer ( OR=1.285, 95% CI 1.079 to 1.530, P=0.005) and PaO 2( OR=0.931, 95% CI 0.895 to 0.970, P=0.001) were the independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. The areas under the ROC curve of chest tightness, D-dimer, PaO 2, and the combination of the three indicators (the combination factor) were 0.728, 0.758, 0.834, and 0.890, respectively. The optimal cut-off value of the combination factor was -3.1, with the sensitivity of 0.824 and the specificity of 0.824. Conclusions:Chest tightness, increased D-dimer and decreased PaO 2 are independent diagnostic indicators for pulmonary embolism in pulmonary tuberculosis patients. It is recommended to perform pulmonary artery computed tomography angiography promptly when the combination factor is higher than -3.1 to determine whether the patient is complicated by pulmonary embolism.
6.Analysis of characteristics of dead on arrival cases in children′s special hospital in the past five years
Li LIU ; Yujuan HUANG ; Tingting CHEN ; Qin XIONG ; Lei SHEN
Chinese Pediatric Emergency Medicine 2021;28(5):367-370
Objective:To analyze the causes, influencing factors and trends of dead on arrival cases in children′s Hospital in the past 5 years, aiming to provide direction and basis for reducing the dead on arrival cases of children.Methods:We collected the dead on arrival cases in the department of emergency at Shanghai Children′s Hospital from January 2015 to December 2019, classifed and analysed the gender, age, native place, death season, time of death, and possible causes of death, and then studied the correlation between above factors and the cases.Results:A total of 151 dead on arrival cases were collected.The annual number decreased year by year, and boys were more than girls in gender.Most of them were infants under 1 year old, and nonlocal children were more than Shanghai native.The above differences were statistically significant, but there was no significant difference in the distribution of death season and death time.In terms of the cause of death, perinatal diseases accounting for 33.8%(51/151), those accompanied with severe underlying diseases accounting for 39.1%(59/151), accidental death accounting for 14.6%(22/151), unexplained deaths accounting for 12.6%(19/151). Those distribution differences were statistically significant( χ2=32.497, P<0.001). Meanwhile, there were statistic differences in gender and age of the cases with severe underlying diseases( χ2=4.898, P=0.027; χ2=32.169, P<0.001), and the year and age distributions of the accidental death cases also had significant differences( χ2=16.636, P=0.002; χ2=14.727, P=0.002). Conclusion:To reduce dead on arrival cases of children, we should do a good job in perinatal health care and screening, reduce premature birth and birth defects, actively conduct propaganda to prevent children′s accidental injuries, popularize medical first aid knowledge, and strengthen children′s transport system.
7.Pressurizing and Stretching Experiment in Arterialized Veins of Rabbits
Yuhao ZHANG ; Pei NIU ; Zhimin ZHANG ; Xiaolong NIU ; Wenzeng SHEN ; Liangchao HAO ; Yujuan ZHOU ; Fulin IU
Journal of Medical Biomechanics 2020;35(2):E156-E162
Objective To establish the model of rabbit vena arterialization, so as to investigate the difference of mechanical parameters between arteries and veins as well as before and after arterialization. Methods Twenty-four rabbits were randomly divided into experimental group (n=12) and control group (n=12). By establishing the rabbit vena arterialization model for experimental group, the arterial blood could flow into the veins. After model creation, the vein would be removed 4 weeks after surgery. In the meantime, the external jugular veins and cephalic arteries extracted from control group were acquired. Compressive pressurizing and stretching tests on all vessels were conducted at the same time(including arteries, veins and arterialized veins). Observation was supported by HE staining and immune tissue chemical techniques. Results There were no deaths among the 24 rabbits, with unobstructed blood flow in veins. With the increase of intravascular pressure, the outer diameter of veins changed at first and then stabilized at a fixed value. The elasticity of veins was worse than that of arteries. The external diameter of veins increased rapidly with internal pressure of veins increasing and reached its extreme elasticity. Comparatively, the elasticity of arteries increased slowly. HE staining results showed that thickness of the vascular wall was thinner, while it became thicker after vena arterialization. After vena arterialization, proliferating cell nuclear antigen(PCNA) and α-actin showed positive results. It further proved that proliferation existed among smooth muscle cells, and veins showed the tendency of restenosis again. The elasticity of veins after transplantation into the arterial system was improved compared with that before transplantation. Conclusions Accompanied by the increasing pressure, the vein could reach its elasticity extremity faster than the artery. Under such a long-term high pressure, vein intima was vulnerable. After vena arterialization, with the gradual thickening of vein intima, the tendency of vessel restenosis was obvious, and the elasticity of veins has been improved after transplantation.
8.Clinical characteristics of 10 cases of brucellosis in Huzhou City, Zhejiang Province
Yujuan SHEN ; Jianfeng ZHONG ; Zhaowei TONG ; Longqi ZHANG ; Weihong WANG
Chinese Journal of Endemiology 2020;39(11):821-824
Objective:To analyze the clinical characteristics of brucellosis in Huzhou City, Zhejiang Province, so as to provide basis for clinicians to improve their understanding and diagnosis and treatment of brucellosis.Methods:The data of 10 cases of brucellosis in Huzhou Central Hospital from 2014 to 2019 were collected, and the epidemiological characteristics, clinical manifestations, laboratory examination results, imaging examination results, etiological examination results, diagnosis and treatment process and prognosis were summarized and analyzed by retrospective analysis method.Results:Among the 10 patients with brucellosis, 6 were males and 4 were females, aged from 32 to 71 years old, and 9 patients had a history of contact with goat, and one case was considered as food borne infection. The main clinical manifestations were fever (10 cases), sweating (2 cases), joint pain (7 cases). In imaging examination, 5 cases with splemomegaly, 1 cases with lymphadenopathy, 3 patients presented with spondylitis (including 1 case with paravertebral abscess) and 2 cases with prostatitis; Brucella was cultured in 2 patients and Brucella antibodies were positive in 10 patients. The treatment drugs included doxycycline, rifampicin and levofloxacin in two or three combinations. Eight patients were followed up for more than 1 month to more than 5 years, and the prognosis was good. Conclusions:Brucellosis lacks specificity in clinical features. Most patients with brucellosis are associated with osteoarthritis or urinary system diseases. Physicians should be vigilant, combine epidemiological features, conduct laboratory tests in time, and diagnose as soon as possible.
9.Clinicopathological characteristics and risk factors of female breast hyperplasia in different age groups
Fei WANG ; Shuohao SHEN ; Liyuan LIU ; Qi WANG ; Hongchuan JIANG ; Lisong TENG ; Zhimin FAN ; Cuizhi GENG ; Feng JIN ; Lili TANG ; Jianguo ZHANG ; Xiang WANG ; Shu WANG ; Xuening DUAN ; Lixiang YU ; Fei ZHOU ; Yujuan XIANG ; Shuya HUANG ; Chao ZHENG ; Qinye FU ; Dezong GAO ; Liang LI ; Zhongbing MA ; Qiang ZHANG ; Zhigang YU
Chinese Journal of General Surgery 2019;34(3):235-239
Objective To analyze clinicopathological characteristics and the potential risk-related factors of female breast hyperplasia in different age groups.Method From Jan 2015 to Dec 2016,patients diagnosed with breast hyperplasia in 12 hospitals were evaluated.All patients completed the self-designed questionnaires on women'health,including basic demographic information,clinic examination information,radiologic information and pathologic results.The patients were divided into a young group (< 45 years old) and an elderly group (from 45 to 75 years old).Results There were 3 684 cases of breast hyperplasia,including 2 291 cases in young group and 1 393 cases in elder group,respectively Clinically breast pain type were most commonly observed in both young and older groups (50.3% vs.42.7%,P < 0.001).While pathological research based on biopsy showed that breast adenopathy were the most common changes in both groups (67.9% vs.61.7%,P <0.001).More breast cancer cases were identified in elder group than that in young group,especially in clinically lump type patients (9.4% vs.4.2%,P < 0.001).Compared with elder group,patients in young group have different distribution characteristics regarding to fertility factors,lifestyle factors and psychology scale including anxiety and depression.Conclusion Distributions of clinicopathological characteristics and risk factors of female breast hyperplasia differ across different age groups.
10.Cyclospora cayetanensis infections among diarrheal outpatients in Shanghai: a retrospective case study.
Yanyan JIANG ; Zhongying YUAN ; Guoqing ZANG ; Dan LI ; Ying WANG ; Yi ZHANG ; Hua LIU ; Jianping CAO ; Yujuan SHEN
Frontiers of Medicine 2018;12(1):98-103
Cyclospora cayetanensis is a foodborne and waterborne pathogen that causes endemic and epidemic human diarrhea worldwide. A few epidemiological studies regarding C. cayetanensis infections in China have been conducted. During 2013, a total of 291 stool specimens were collected from patients with diarrhea at a hospital in urban Shanghai. C. cayetanensis was not detected in any of the stool specimens by traditional microscopy, whereas five stool specimens (1.72%, 5/291) were positive by PCR. These positive cases confirmed by molecular technology were all in the adult group (mean age 27.8 years; 2.94%, 5/170) with watery diarrhea. Marked infection occurred in the rainy season of May and July. Sequence and phylogenetic analyses of the partial 18S rRNA genes of C. cayetanensis isolated showed intra-species diversity of this parasite. This study showed, for the first time, that C. cayetanensis is a pathogen in outpatients with diarrhea in Shanghai, albeit at a low level. However, the transmission dynamics of this parasite in these patients remain uncertain.
Adolescent
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Adult
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China
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epidemiology
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Cyclospora
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genetics
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isolation & purification
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Cyclosporiasis
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epidemiology
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Diarrhea
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etiology
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parasitology
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Feces
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parasitology
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Female
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Humans
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Male
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Middle Aged
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Outpatients
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Phylogeny
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Polymerase Chain Reaction
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RNA, Ribosomal, 18S
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analysis
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Retrospective Studies
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Young Adult

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