1.Production and identification of polyclonal antibody against HSF1 in sheep and immunohistochemical analysis
Wenyu FAN ; Lanying GU ; Qinchuan ZHANG ; Shiyi LI ; Yanbing ZHANG ; Yanming SUN
Chinese Journal of Veterinary Science 2025;45(11):2394-2404
Heat shock factor 1(HSF1)is a core transcription factor in cellular stress response and protein homeostasis maintenance,and is widely involved in biological processes such as cell growth,differentiation,and metabolism.This study aims to design antigenic peptides of the sheep HSF1 protein and prepare polyclonal antibodies through bioinformatics analysis and immunologi-cal techniques,and verify their application effects in sheep lung and testis tissues.Firstly,the se-quence,structure,and evolution of the sheep HSF1 gene were analyzed using bioinformatics meth-ods,and key antigenic epitopes were predicted.Then,suitable peptide fragments were selected for synthesis,emulsified with Freund's complete adjuvant or incomplete adjuvant,and used to immu-nize New Zealand rabbits.The specificity and effectiveness of the antibodies were verified by indi-rect ELISA,Western blot,and immunohistochemistry(IHC)experiments.The results showed that the HSF1 protein is 564 amino acids(aa)in length,with a molecular weight of 60.76 kDa,a theo-retical isoelectric point(pI)of 5.32,and is hydrophilic and unstable.Phylogenetic tree analysis revealed that HSF1 is highly conserved among most members of the Bovidae family,as well as in humans,mice,rats,and other species,but with local site differences.Sheep HSF1 is most closely related to goat HSF1,followed by Canadian bighorn sheep and goral.The phosphorylation sites of sheep HSF1 are predominantly concentrated in the middle and C-terminal regions,which is largely consistent with the predicted results for human HSF1,though some local differences exist.The prepared polyclonal antibodies exhibited a titer of over 1:8000 in recognizing the sheep HSF1 pro-tein,and Western blot experiments confirmed clear bands with consistent molecular weight,indica-ting high specificity of the antibodies.Furthermore,the expression and specific distribution of HSF1 were detected in sheep lung and testis tissues.This study successfully prepared polyclonal antibodies based on the sheep HSF1 antigen peptide and,for the first time,detected the immuno-histochemical distribution of HSF1 in the testis and lung tissues of Argali hybrid sheep.This pro-vides an important tool for further exploring the physiological role of HSF1 in Argali hybrid sheep and its potential applications in stress response,disease prevention,and treatment.
2.Development process of family resilience in children with congenital pseudarthrosis of the tibia based on grounded theory
Lan YIN ; Jianhui XIE ; Jialing LONG ; Le XU ; Lanying WANG ; Yaqi OUYANG ; Yinzhi YI ; Can GU
Chinese Journal of Modern Nursing 2025;31(12):1599-1604
Objective:To explore the development process of family resilience in children with congenital pseudarthrosis of the tibia (CPT) and to understand the long-term challenges and coping strategies that CPT imposes on the families of affected children.Methods:This study combined purposive sampling and theoretical sampling. It selected 15 caregivers of CPT children from Hunan Children's Hospital for semi-structured interviews. Grounded theory was used to analyze the interview results.Results:The development of family resilience in CPT children's families occurred in three stages: pre-formation stage, formation stage, and maintenance stage. In facing negative emotions and family challenges, caregivers first needed to rebuild their inner beliefs. They then adjusted the family organizational model, adopted open and inclusive communication, and actively sought external support to foster the development of family resilience. Ultimately, caregivers were able to self-regulate their emotions, accumulate caregiving experience, and begin to shift their life perspective.Conclusions:The development of family resilience in CPT children's families is a dynamic, multi-stage process with interactions of multiple factors. Healthcare providers should offer professional health guidance according to the different stages of family development. Moreover, the government and schools should increase their attention and support, working together with families and healthcare providers to enhance family resilience for children with CPT.
3.Production and identification of polyclonal antibody against HSF1 in sheep and immunohistochemical analysis
Wenyu FAN ; Lanying GU ; Qinchuan ZHANG ; Shiyi LI ; Yanbing ZHANG ; Yanming SUN
Chinese Journal of Veterinary Science 2025;45(11):2394-2404
Heat shock factor 1(HSF1)is a core transcription factor in cellular stress response and protein homeostasis maintenance,and is widely involved in biological processes such as cell growth,differentiation,and metabolism.This study aims to design antigenic peptides of the sheep HSF1 protein and prepare polyclonal antibodies through bioinformatics analysis and immunologi-cal techniques,and verify their application effects in sheep lung and testis tissues.Firstly,the se-quence,structure,and evolution of the sheep HSF1 gene were analyzed using bioinformatics meth-ods,and key antigenic epitopes were predicted.Then,suitable peptide fragments were selected for synthesis,emulsified with Freund's complete adjuvant or incomplete adjuvant,and used to immu-nize New Zealand rabbits.The specificity and effectiveness of the antibodies were verified by indi-rect ELISA,Western blot,and immunohistochemistry(IHC)experiments.The results showed that the HSF1 protein is 564 amino acids(aa)in length,with a molecular weight of 60.76 kDa,a theo-retical isoelectric point(pI)of 5.32,and is hydrophilic and unstable.Phylogenetic tree analysis revealed that HSF1 is highly conserved among most members of the Bovidae family,as well as in humans,mice,rats,and other species,but with local site differences.Sheep HSF1 is most closely related to goat HSF1,followed by Canadian bighorn sheep and goral.The phosphorylation sites of sheep HSF1 are predominantly concentrated in the middle and C-terminal regions,which is largely consistent with the predicted results for human HSF1,though some local differences exist.The prepared polyclonal antibodies exhibited a titer of over 1:8000 in recognizing the sheep HSF1 pro-tein,and Western blot experiments confirmed clear bands with consistent molecular weight,indica-ting high specificity of the antibodies.Furthermore,the expression and specific distribution of HSF1 were detected in sheep lung and testis tissues.This study successfully prepared polyclonal antibodies based on the sheep HSF1 antigen peptide and,for the first time,detected the immuno-histochemical distribution of HSF1 in the testis and lung tissues of Argali hybrid sheep.This pro-vides an important tool for further exploring the physiological role of HSF1 in Argali hybrid sheep and its potential applications in stress response,disease prevention,and treatment.
4.Development process of family resilience in children with congenital pseudarthrosis of the tibia based on grounded theory
Lan YIN ; Jianhui XIE ; Jialing LONG ; Le XU ; Lanying WANG ; Yaqi OUYANG ; Yinzhi YI ; Can GU
Chinese Journal of Modern Nursing 2025;31(12):1599-1604
Objective:To explore the development process of family resilience in children with congenital pseudarthrosis of the tibia (CPT) and to understand the long-term challenges and coping strategies that CPT imposes on the families of affected children.Methods:This study combined purposive sampling and theoretical sampling. It selected 15 caregivers of CPT children from Hunan Children's Hospital for semi-structured interviews. Grounded theory was used to analyze the interview results.Results:The development of family resilience in CPT children's families occurred in three stages: pre-formation stage, formation stage, and maintenance stage. In facing negative emotions and family challenges, caregivers first needed to rebuild their inner beliefs. They then adjusted the family organizational model, adopted open and inclusive communication, and actively sought external support to foster the development of family resilience. Ultimately, caregivers were able to self-regulate their emotions, accumulate caregiving experience, and begin to shift their life perspective.Conclusions:The development of family resilience in CPT children's families is a dynamic, multi-stage process with interactions of multiple factors. Healthcare providers should offer professional health guidance according to the different stages of family development. Moreover, the government and schools should increase their attention and support, working together with families and healthcare providers to enhance family resilience for children with CPT.
5.Influential factors of psychological state and their correlation with nutritional risk in patients undergoing thoracic tumor radiotherapy
Xiaoying SHEN ; Xi YANG ; Yuyan GU ; Fang NIU ; Lanying QIU ; Shuping XIE
Chinese Journal of Primary Medicine and Pharmacy 2022;29(4):538-543
Objective:To investigate the prevalence of anxiety and depression in patients undergoing thoracic tumor radiotherapy, preliminarily evaluate the nutritional risk of patients, and analyze the possible influential factors of anxiety and depression and their correlation with nutritional risk, providing a theoretical basis for further formulating a personalized psychological nursing plan for patients undergoing thoracic tumor radiotherapy.Methods:A total of 121 patients who underwent thoracic tumor radiotherapy between April 2019 and March 2020 in Cancer Hospital of The University of Chinese Academy of Sciences were selected for this study. We evaluated patients' psychological statuses using the Hospital Anxiety and Depression Scale and patients' nutritional risk using the Nutrition Risk Screening. We performed univariate analysis, unconditional logistic regression analysis, and contingency analysis to analyze the factors that affected the psychological status of patients undergoing thoracic tumor radiotherapy and correlate patients' psychological status with nutritional risk.Results:Among 121 patients, 99 (81.8%) patients undergoing thoracic tumor radiotherapy would develop or had developed anxiety, and 42 (34.7%) patients were at the boundary of depression or had depression. Correlation analysis revealed that nutritional risk was correlated with the prevalence of anxiety and depression. Logistic regression analysis showed that surgery, first radiotherapy, and nutritional risk were the factors that affected the prevalence of anxiety. Marital status, location of tumors, surgery, first radiotherapy, and nutritional risk affected the prevalence of depression.Conclusion:Patients undergoing thoracic tumor radiotherapy have serious psychological problems. Nutritional risk is positively correlated with the prevalence of anxiety and depression. Surgery and first radiotherapy are the factors that affect the prevalence of anxiety and depression.
6.The potential role of lung dendritic cells and Th17/regulatory T cells in patients with chronic obstructive pulmonary disease
Lanying ZHANG ; Jie CHEN ; Yi ZHANG ; Yanhui GU ; Ximin RAO ; Yao OUYANG
Chinese Journal of Internal Medicine 2019;58(2):125-132
Objective To explore the role of lung dendritic cells (DCs) and Th17/regulatory T cells (Treg) pathway in the pathogenesis of chronic obstructive pulmonary disease(COPD).Methods COPD patients who received lobectomy from Sep.2015 to Mar.2016 in our hospital were enrolled and classified into non-smoking non-COPD group,smoking without COPD group and COPD group.The expression of CD80,chemokine recepter-6 (CCR6),interleukin-17A (IL-17A) and fork-head transcription factor P3 (FoxP3) were detected by immunohistochemistry (IHC) in lung tissue.Mature DCs (mDCs),immature DCs (imDCs),Th17 cells and Treg cells in lung tissue were detected by flow cytometry (FCM) and the correlation between Th17/ Treg cells with lung function was analyzed.Results (1) The expression of CD80 and FoxP3 in COPD group was decreased,while the expression of CCR6 and IL-17A was increased (P<0.05).(2) The percentage of mDCs and Treg in lung tissue of COPD group was significantly decreased.In contrast,the proportion of imDCs and Th17 cells in COPD group was significantly increased (P<0.05).(3) The imbalance of Th17/Treg ratio in lung tissue was seen in patients with COPD,suggesting the potential mechanism of Th17 cell-mediated proinflammatory response.(4) The percentage of Th17 cells and Th17/Treg ratio in COPD patients was negatively correlated with forced expiratory volume in the first second (FEV1) as a percentage of predicted value (FEV1% pred),forced vital capacity(FVC) as a percentage of predicted value (FVC% pred),FEV1/FVC.On the other hand,the percentage of Treg cells was positively correlated with FEV1% pred,FVC% pred,FEV1/FVC.Conclusions The data in this study demonstrate the maturation disorder of dendritic cells in lung tissue of COPD patients.The imbalance of Th17/Treg ratio suggests that Th17 cell-mediated proinflammatory response may be involved in the pathogenesis of COPD.
7.A study of inductive factors of embryonic stem cells differentiating into cardiac myocytes in vitro
Lanying LIU ; Kun YANG ; Zhengyu ZHU ; Yuchuan LIU ; Xiyong YU ; Jian TANG ; Wei YIN ; Jianquan MA ; Jun GU
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To study cellular and molecular mechanisms of cardiac development associated genes ex- pression and its function during early stage cardiomyogenesis. METHODS: (1 ) Mouse embryonic stem cells (ESC) line D3 culture. (2) Inductive culals of ESC differentiated into cardiac myocytes in vitro.(3) Identification of ESC -derived cardiac myocytes: RNA isolation; synthesis of specific primer and RT - PCR; Label of RT - PCR products with [? - 32P] dATP as probes, purifyed by sephadex G - 50 columns, determined the yield of DNA. RNA dot hy- bridization. RESULTS: 80% of ESC differentiated into cardiomyocytes by improved conditional medium. Cardiomy- ocytes contraCted in a synchronous manner. The results of RT - PCR and RNA blot showed that cardiac genes were expressed abundantly and specifically during the early cardiomyogenesis. CONCLUSIONS: ESC were able to be dif- ferentiate into cardiomyocytes. Different concentrations and components of RA, DMSO and FCS affected ESC car- diomyogenesis in de. The optimal result obtained was from the conditional medium, a mixturce of 2 nmol/L retinoic acid (RA), 0.6% dimethyl sulfoxide (DMSO) and 20% fend calf serum (FCS).

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