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1.
Chinese Pharmacological Bulletin ; (12): 2346-2353, 2023.
Article in Chinese | WPRIM | ID: wpr-1013677

ABSTRACT

Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress. Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly, with six mice in each group. The mice in normal group or high-fat diet group were fed with regular or high-fat diet continuously for five months. The mice were weighed, anesthetized, and euthanized to collect testicular and epididymal tissue for analysis. The testicular tissue was weighed and their indices were calculated. Epididymal tissue was collected for semen analysis. The morphological alterations of testicular tissue were observed using hematoxylin-eosin ( HE ) staining. The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calculated. The expression levels of apoptosis and endoplasmic reticulum stress-related proteins in testicular tissue were detected by Western blot. The protein expression and localization of GRP78 in testicular tissue were further detected by immunofluorescence. Results The results showed that compared to the normal group, the high-fat diet group had a significant increase in body weight, a significant decrease in testicular index, sperm concentration, and sperm vability, loose arrangement of germ cells, significant thinning of the seminiferous epithelium, no significant change in the diameter of seminiferous tubules, a significant increase in germ cell apoptosis , with an increased apoptosis index, and significant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expression. The expression levels of GRP78 , p-IREl, XBP1, and ATF6a proteins were significantly up-regulated, while p-PERK, p-eIF2a, ATF4 protein expression showed no significant changes. Immunofluorescence results further showed a significant increase in the expression of GRP78 protein in the testicular tissue,with no significant changes in the expression location. Conclusions High-fat diet can induce the apoptosis of mouse testicular germ cells, and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 98-103, 2020.
Article in Chinese | WPRIM | ID: wpr-872797

ABSTRACT

Objective::To observe the efficacy of Wenfei Jianpi decoction on moderate to severe persistent allergic rhinitis (AR) with Lung Qi deficiency and cold syndrome and its effect on substance P (SP) in nasal secretions and peripheral blood proinflammatory factors. Method::One hundred and sixty-six patients were randomly divided into control group and observation group by random number table. Patients in control group got budesonide nasal spray, 2 times/days, and cetirizine hydrochloride tablets, 10 mg/days. And patients in observation group got Wenfei Jianpi decoction for oral, 1 dose/day. A course of treatment was 8 weeks. Before and after treatment, nasal obstruction, itching, sneezing, runny nose, itchy eyes, tears and nasal sign were scored, nasal airway resistance was detected, and rhinoconjunctivits quality of life questionnaire (RQLQ) were scored. Before and after treatment, levels of SP immunoglobulin E (IgE) in peripheral blood, interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected, and eosinophil (EOS) were calculated. Result::By rank sum test, the clinical efficacy in observation group was better than that in control group (Z=2.073, P<0.05). Before and after treatment, scores of total score of nasal symptoms, ocular symptoms and subjective symptoms and nasal sign score were all lower than those in control group (P<0.05). Except sleep and emotion, scores of other dimensions and the total scores of RQLQ were all lower than those in control group (P<0.05). EOS in observation group was less than that in control group (P<0.05). And nasal resistance declined in observation group (P<0.05), and levels of IgE, SP, IL-4, IL-5, IL-6, TNF-α and nasal resistance were lower than those in control group (P<0.05, P<0.01). Conclusion::Wenfei Jianpi decoction can be used to treat moderate to severe persistent allergic rhinitis by relieving subjective symptoms and signs, improving quality of life, inhibiting inflammatory factors and reducing allergic reaction.

3.
National Journal of Andrology ; (12): 387-391, 2013.
Article in Chinese | WPRIM | ID: wpr-350894

ABSTRACT

<p><b>OBJECTIVE</b>To construct a mammalian expression plasmid of the BC022687 gene and investigate the expression and localization of the fusion protein in Chinese hamster ovary (CHO) cells.</p><p><b>METHODS</b>The BC022687 coding sequence was amplified by polymerase chain reaction (PCR) and subcloned into the pEGFP-C1 vector carrying the gene of green fluorescence protein (GFP). After the target region was sequenced, the recombinant plasmid was transfected into CHO cells, and its expression in the CHO cells was determined by Western blot. The localization of GFP-tagged BC022687 in the CHO cells was observed by laser scanning confocal microscopy.</p><p><b>RESULTS</b>BC022687 was successfully cloned into the mammalian expression vector pEGFP-C1, with the restriction fragment length of 950 bp. The expression of the fusion protein was confirmed, with the relative molecular weight of 64 000. The GFP-tagged BC022687 protein was mainly localized in the cytoplasm, and also presented in the centrioles in the transfected CHO cells.</p><p><b>CONCLUSION</b>The successful construction of the plasmid expressing BC022687 in CHO cells has laid a foundation for further studies on the role of this protein in ciliogenesis.</p>


Subject(s)
Animals , Cricetinae , Male , Mice , CHO Cells , Centrosome , Metabolism , Cilia , Metabolism , Cricetulus , DNA, Complementary , Genetic Vectors , Plasmids , Recombinant Fusion Proteins , Genetics , Transfection
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