1.Expression of N-methyl-D-aspartic acid receptor and endoplasmic reticulum stress related pathway proteins in brain tissue of fluorosis rats
Chun YANG ; Jianxia WEN ; Jianglong FENG ; Zhizhong GUAN ; Na WEI
Chinese Journal of Tissue Engineering Research 2024;28(7):1070-1075
BACKGROUND:Previous studies have shown that N-methyl-D-aspartic acid receptor(NMDA)receptors are associated with fluorine,but the role in fluoride-induced endoplasmic reticulum stress remains unclear. OBJECTIVE:To observe the changes of excitatory neurotransmitter NMDA receptor and endoplasmic reticulum stress IRE1α-ASK1-JNK pathway protein expression in brain tissue of rats with experimental fluorosis,and to investigate the pathogenesis of neurological injury in fluorosis by giving NMDA receptor inhibitor to SH-SY5Y cells. METHODS:(1)Animal model:18 1-month-old SD rats were randomly divided into control group(drinking water fluoride content<0.5 mg/L),low fluoride group(drinking water fluoride content 10.0 mg/L)and high fluoride group(drinking water fluoride content 100.0 mg/L),with 6 rats in each group,half of each sex.After 6 months of fluoride intake,the rats were observed for the occurrence of dental fluorosis,and the 24-hour urinary fluoride content was measured.After anesthesia and euthanasia,the brain tissue of rats was taken to observe the pathological changes.Western blot assay was used to detect NMDA receptors and IRE1α,ASK1 and JNK protein expression in the brain tissue.(2)Cell model:SH-SY5Y cells were cultured in vitro and treated with sodium fluoride at final concentrations of 0.3 mmol/L and 3 mmol/L.The fluoride-stained cells were interfered with 10 μmol/L NMDA receptor antagonists Ifenprodil and MK-801 to observe the relevant protein changes. RESULTS AND CONCLUSION:(1)The incidence of dental fluorosis and urinary fluoride level in rats in the high fluoride group were significantly higher than that in the control and low fluoride groups(P<0.05).(2)Compared with the control group,the cytoplasm of neuronal cells in the CA3 area of the hippocampus in the low fluoride group was slightly more basophilic,while the neuronal cells in the CA3 area of the high fluoride group were disorganized,with increased basophilicity and some of the nuclei solidified.(3)In rat brain tissue,the expressions of NR2A in the high fluoride group and NR2B in the low fluoride group were significantly higher compared with the control group(P<0.05),and NR2B,IRE1,ASK1,and p-JNK protein expression levels were increased in the high fluoride group compared with the control and low fluoride groups(P<0.05).(4)In SH-SY5Y cells,NR1,NR2A and NR2B protein expressions were significantly increased in the high fluoride group compared to the control group(P<0.05).The protein levels of NR1 and NR2A were significantly reduced in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).NR2B protein expression was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(5)In SH-SY5Y cells,IRE1,ASK1,and p-JNK protein expression was significantly higher in the high fluoride group compared with the control group(P<0.05),while ASK1 and p-JNK protein expressions were significantly decreased in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).IRE1 protein level was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(6)It is concluded that excessive fluorine intake activates NMDA receptors in the central nervous system,causing increased expression of endoplasmic reticulum stress IRE1α,ASK1,and p-JNK proteins,and the use of NMDA receptor inhibitors has a mitigating effect on endoplasmic reticulum stress caused by fluorosis.
2.Effects of different doses of fluoride on expression of IRE1α-ASK1-JNK proteins in rat renal cortex
Jianxia WEN ; Li HE ; Jianglong FENG ; Na WEI
Chinese Journal of Endemiology 2022;41(2):100-104
Objective:To investigate the expression and significance of endoplasmic reticulum stress apoptosis pathway related proteins in renal cortex of rats with chronic fluorosis.Methods:Twenty four healthy SD rats were divided into 4 groups (6 rats/group, half male and half female) according to their body mass (100 - 120 g) by random number table method, rats in control group drank tap water (fluoride content < 0.5 mg/L), and in low, medium and high fluoride groups drank tap water with fluoride content (sodium fluoride) of 10, 50 and 100 mg/L, respectively. After 180 days of feeding, dental fluorosis was examined, 24-hour urine sample was collected and the content of fluoride in urine was detected by fluoride ion selective electrode method. Renal tissue was taken after anesthesia, and the pathological changes of renal cortex were observed by hematoxylin-eosin (HE) staining. The expressions of endoplasmic reticulum stress apoptosis pathway related proteins [inositol-requiring enzyme 1α (IRE1α), apoptosis signal-regulating kinase 1 (ASK1), C-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK)] were determined by immunohistochemical staining in rat renal cortex.Results:No dental fluorosis was found in control group. The incidence of dental fluorosis in low, medium and high fluoride groups were 2/6, 5/6 and 6/6, respectively. Compared with control group [(5.707 ± 1.190) mg/L], the urinary fluoride in low, medium and high fluoride groups [(17.028 ± 3.006), (34.378 ± 12.045), (94.759 ± 31.773) mg/L] was significantly higher ( P < 0.05), and the urinary fluoride in high fluoride group was higher than that in low and medium fluoride groups ( P < 0.05). HE staining showed that, compared with control group, the cell volume of renal tubules and glomeruli in medium and high fluoride groups increased, the cells arranged closely, and the eosinophilia of the cytoplasm increased. The immunohistochemical staining results showed that there was no significant difference in the expression of JNK protein in rat renal cortex between control group and low, medium and high fluoride groups ( F = 0.07, P > 0.05). The expressions of IRE1α, ASK1 and P-JNK proteins in rat renal cortex in high fluoride group were higher than those of control, low and medium fluoride groups ( P < 0.05), and the expressions of IRE1α and ASK1 proteins in medium fluoride group were significantly higher than those in control and low fluoride groups ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to renal cortex injury in rats, and the mechanism of injury may be related to the activation of IRE1α-ASK1-JNK endoplasmic reticulum stress apoptosis pathway.
3.A survey on human resources of dentists who deal with periodontal disease in Beijing
Xianghui FENG ; Jianxia HOU ; Jingran ZHANG ; Rongsen LIU ; Fengqiu ZHANG ; Jiang LIN ; Xiangying OUYANG ; Yi LIU ; Zuomin WANG ; Qingxian LUAN
Chinese Journal of Stomatology 2021;56(11):1109-1114
Objective:To investigate the status quo of human resources of dentists who deal with periodontal disease in Beijing area through an online survey, which may hopefully provide a preliminary basis for the decision-making of administrative departments and the formulation of periodontal professional development plan.Methods:The dentists who deal with periodontal disease at least half a day per week in Beijing area were investigated. A questionnaire was designed by the chairman of the Periodontology Committee of Beijing Stomatological Association. The questionnaire was sent to and finished by the dentists via "WenJuanXing" online survey software. The contents of the survey included general condition, the property of practice unit, title and position of the dentist, membership of professional society, time and content of periodontal treatment, adoption of new technology and new method of periodontal therapy during the past one year, status of periodontal treatment in the local population and reasons, understanding and influencing factors of periodontal professional development.Results:A total of 1 255 dentists completed the survey, who came from all 16 districts in Beijing, mainly Haidian, Chaoyang, Dongcheng and Xicheng Districts [The total percentage of these four main districts was 70.3% (882/1 255)]. The mean age of the dentists was (36.1±8.3) years. Among the dentists, 71.1% (892/1 255) were females, 88.1% (1 106/1 255) got a Bachelor′s degree or above. It was estimated that 35.4% (444/1 255) of the dentists had received standardized periodontal training ever. The percentage of dentists carrying out new technology in the past one year was as high as 68.1% (855/1 255). There were only 163 periodontal specialists (13.0%) out of the dentists in the survey. Only 15.9% (200/1 255) of the dentists routinely performed periodontal surgery. The majority of the dentists [82.8% (1 039/1 255)] were from the state-owned hospitals. Fifty-four point seven percemt (686/1 255) of the dentists thought that lack of knowledge was the main reason why the general public failed to receive periodontal treatment. As for the biggest bottleneck affecting periodontal professional development, fifty-one point zero percent (640/1 255) of the dentists attributed it to the public neglect.Conclusions:The periodontal practitioners in Beijing are young, highly educated, unevenly distributed in 16 districts and mostly females. State-owned oral health institutions are an important force in periodontal diagnosis and treatment services in Beijing. The number of periodontal specialists need to be improved. Promotion of standardized periodontal surgery and the popularization of healthcare knowledge on periodontal disease should also be the focus in the future.
4.Expression of ERAP1, TNFR-Ⅰ, TNFR-Ⅱ in psoriasis vulgaris
Jianxia CHEN ; Xinmei LIU ; Jianyong LIU ; Peng WANG ; Xiaomin CHEN ; Qingyu MA ; Yanyan FENG
Journal of Chinese Physician 2020;22(10):1488-1492
Objective:To explore the possible role of endoplasmic reticulum aminopeptidase 1 (ERAP1), tumor necrosis factor-α receptor Ⅰ (TNFR-Ⅰ), tumor necrosis factor-α receptor Ⅱ (TNFR-Ⅱ) in the pathogenesis of psoriasis vulgaris.Methods:15 cases of normal skin and 20 cases of psoriasis vulgaris in Xinjiang Uygur Autonomous Region People's hospital were enrolled. Immunohistochemical staining techniques were used to detect the expression of ERAP1, TNFR-Ⅰ, TNFR-Ⅱ protein in the two groups. Statistical analysis was then performed to compare the difference in expression between the two groups.Results:⑴ In normal skin , ERAP1 is mainly expressed in the basal layer of the epidermis. In the psoriatic vulgaris lesions, ERAP1 was diffusely positive. The expression of ERAP1 in the psoriatic lesions was stronger than that of normal tissues, with stastically significant difference ( Z=-4.170, P<0.05). ⑵ TNFR-Ⅰ is diffusedly expressed in the normal skin tissues. However, in the epidermis of psoriatic lesions, the expression of TNFR-Ⅰ in the spinous layer was stronger than that of the basal layer (χ 2=17.740, P<0.05). ⑶ In the normal skin, TNFR-Ⅱ was not expressed; in the psoriatic lesions, TNFR-Ⅱ was diffusely expressed in the basal cell layer and acanthosis of the epidermis in varying degrees, and the difference between normal and psoriatic skin was statistically significant (χ 2=17.500, P<0.001). ⑷ The expression of ERAP1 in the epidermis of psoriasis vulgaris was negatively correlated with TNFR-Ⅰ ( rs=-0.662, P=0.001). There was no significant correlation between ERAP1 and TNFR-Ⅱ ( rs=0.343, P=0.139). Conclusions:ERAP1, TNFR-Ⅰ, and TNFR-Ⅱ may play important roles in the pathogenesis of psoriasis vulgaris. They may interact with each other to regulate the proliferation and apoptosis of keratinocyte in order to maintain the abnormal proliferation and benign proliferation of psoriasis vulgaris epidermis.
5.Preparation of colloidal gold test strip against human SMCY antigen
Zhiyuan AN ; Huaigu ZHOU ; Jianxia WANG ; Xiaoyan FENG ; Dong ZHAO
Chinese Journal of Forensic Medicine 2018;33(1):51-54
Objective To establish a rapid method for the detection of SMCY antigen. Methods To Use the technology of colloidal gold immunochromatography with double antibody sandwich assay, the gold labeled pad was coated with colloidal gold labeled SMCY rabbit polyclonal antibody, colloidal gold strip was made for detection of serum, which include 12 serum samples of pig, cattle, dog, chicken and mice and 50 serum samples of human. Results The colloidal gold test strips showed obvious specificity in human and common animal sera and could distinguish between male and female sera. Conclusion This method can be used to identify the serum of women and men, and has certain species specificity, which provides a direction for forensic science to rapidly identify gender of human samples.
6.Effects of continuous nursing care on postoperative urinary incontinence in patients with prostate cancer
Haiyan LI ; Yao FENG ; Jianxia CHEN ; Haiwei ZHANG ; Rongrong WU ; Zhiliang WENG ; Haihong JIANG ; Wei CHEN
Chinese Journal of Modern Nursing 2018;24(3):339-343
Objective To explore the effects of continuous nursing on postoperative urinary incontinence in patients with prostate cancer, so as to provide clinical evidence to improve the quality of postoperative and extended nursing care.Methods A total of 120 patients with prostate cancer underwent radical prostatectomy in the First Affiliated Hospital of Wenzhou Medical University from January 2016 to January 2017, who met the inclusion criteria were recruited in the research. The patients were randomly divided into experimental group and control group by sortition randomization method, with 60 cases in each. The patients in control group were given routine discharge health education, and the experimental group received continuous nursing on the basis of routine discharge nursing. Three months after discharge, the incidence, duration, frequency and quantity of urinary incontinence in the two groups were recorded by the International Consultation on Incontinence Questionnaire Short Form (ICI-Q-SF). A self-designed prostate cancer postoperative-related knowledge questionnaire, which evaluated the patients understanding prostate cancer care, was completed by the patients of the two groups. A self-designed satisfaction questionnaire was used to evaluate and compare the satisfaction of the two groups of patients to the quality of nursing.Results The incidence rate and the duration of postoperative urinary continence in the experimental group [16.7%,(4.1±2.8) d] were significantly lower than those in the control group (χ2=9.076,t=-2.630;P<0.05). After 3 months of discharge, the score of related knowledge and the satisfaction of nursing work in the experimental group were both higher than those of the control group, and the differences were statistically significant (P<0.05).Conclusions Continuous nursing care for discharged patients after prostatectomy can enhance patients' knowledge of disease, effectively reduce the incidence of postoperative incontinence, reduce urinary incontinence duration, and improve patients' satisfaction with nursing quality.
7.Qualitative study on psychological experience in elderly patients with prostatic cancer after castration
Haiyan LI ; Haihong JIANG ; Yao FENG ; Jianxia CHEN ; Rongrong WU ; Wei CHEN
Chinese Journal of Modern Nursing 2018;24(23):2777-2779
Objective To explore the real psychological experience in elderly patients with prostatic cancer after castration and to offer reference for formulating targeted interventions.Methods Totally 15 elderly prostatic cancer patients who received castration and total androgen blockade in the First Affiliated Hospital of Wenzhou Medical University from October 2014 to June 2016 were selected by convenient sampling and interviewed intensively.The data were analyzed by Colaizzi's seven-step method for qualitative studies.Results Totally 3 topics were summarized:the sense of uncertainty in elderly prostatic cancer patients after castration;changes in social adaptation;self-image disorder.Conclusions Various psychological crises,especially depression,are experienced in elderly prostatic cancer patients after castration.Therefore,continuous and extensive support is required for elderly prostatic cancer patients after castration.
8.The application of the cloning and expression of SMCY gender-specific fusion antigens with the corresponding polyclonal antibodies
Zhiyuan AN ; Lu TIAN ; Jianxia WANG ; Xiaoyan FENG ; Jianping TANG ; Xin CHEN ; Ronghua CHEN ; Gang BI ; Xiong XIAO
Chinese Journal of Forensic Medicine 2017;32(4):375-378
Objective with the advantages of rapidity in detection protein, We selected the gender-specific amino acid sequence based on human SMCY and SMCX, cloned and expressed SMCY gender-specific fusion antigens. The rabbits were immunized with purified antigens to obtain the polyclonal antibodies. A new method was established for rapidly sex identification of forensic evidence samples by detecting SMCY antigens with the corresponding polyclonal antibodies. Methods We found three differential fragments by analyzing the sequence of human SMCY and SMCX. Then we cloned this three fragments and ligated as a new recombinant.This SMCY gender-specific fusion antigen gene was sub-cloned into pET-28a and expressed in Escherichia coli. The fusion antigen was purified by Ni-NTA column. The rabbits were immunized with purified antigen to produce the specific polyclonal antibodies.The reactivity of the polyclonal antibody was evaluated by ELISA and Western blotting. We developed a colloidal gold test strip for detecting the gender of human samples. Results We successfully selected gender-specific amino acid sequence, the SMCY gender-specific fusion antigen was expressed by prokaryotic expression and the polyclonal antibody was prepared by immunizing rabbit. The results of colloidal gold strip tests showed that there is a significant difference between male and female serums. Conclusion The results showed that the SMCY gender-specific fusion antigen could be recognized by the polyclonal antibody.The colloidal gold strip tests made by SMCY gender-specific fusion antigens and the corresponding polyclonal antibodies could be used for rapidly determining the gender of forensic evidence samples.
9.The role of SR-BⅡ in the oxidized low density lipid induced foam cell formation promoted by porphyromonas gingivalis
Xiaoli HE ; Dongyu LIANG ; Feng LIU ; Jianxia CHEN ; Lijun LUO
Journal of Practical Stomatology 2017;33(3):306-311
Objective:To investigate the role of scavenger receptor BⅡ(SR-BⅡ) in the oxidized low density lipid(ox-LDL) induced foam cells formation promoted by porphyromonas gingivalis(P.g).Methods:Peritoneal macrophages were stimulated with P.g and ox-LDL,and then foam cells formation were checked.The expression of SR-BⅡ was detected by Western blot and real-time PCR.Next,siRNA targeting SR-BⅡ was used to detect the change of foam cells formation.Results:After being stimulated with P.g and ox-LDL,the foam cells formation was significantly increased.During the process of foam cells formation,P.g infection increased the expression of SR-BⅡ.And the knockdown of SR-BⅡ by siRNA significantly reduced the foam cells formation.Conclusion:P.g infection can increase the expression of SR-BⅡ and the regulation of SR-BⅡ expression can change the foam cells formation.
10.Efficacy of alendronate sodium combined with dietary and exercise intervention in postmenopausal osteoporosis
Jing GAO ; Chunying MAO ; Jianxia ZHANG ; Hui JIN ; Yujin FENG ; Xiaojun RAO
Chinese Journal of Biochemical Pharmaceutics 2017;37(1):163-164,168
Objective To evaluate the efficacy of alendronate sodium combined with dietary and exercise intervention in postmenopausal osteoporosis. Methods 88 postmenopausal osteoporosis patients were enrolled and treated with alendronate sodium and Vitamin D combined with dietary and exercise intervention for 12 months. Bone mineral density (BMD) of lumber spine and femur neck were measured before and after the treatment. Results Medication combined with dietary and exercise intervention significantly increased bone mineral density of postmenopausal osteoporosis patients, and the serum levels of bone alkaline phosphatase (BLAP), alkaline phosphatase (AKP) and tartrate resistant acid phosphatase (TRACP) were significantly higher than those pre-treatment, with significant differences (P<0.05). Conclusion Compared to simple medication treatment, medication combined with dietary and exercise intervention is more effective to enhance medication efficacy and BMD level.

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