Objective:To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase (HO-1) signaling pathway in the attenuation of endotoxin-induced acute lung injury (ALI) by hydromorphone and the relationship with Golgi apparatus stress (GA stress) in mice.Methods:Eighteen SPF wild-type (WT) and 18 Nrf2 knockout (Nrf2 KO) male C57BL/6J mice, aged 6-8 weeks, weighing 18-20 g, were divided into 3 groups ( n=6 each) using a random number table method: control groups (WT+ Con group, Nrf2 KO+ Con group), ALI groups (WT+ ALI group, Nrf2 KO+ ALI group) and ALI+ hydromorphone groups (WT+ ALI+ HM group, Nrf2 KO+ ALI+ HM group). ALI was induced by injecting lipopolysaccharide (LPS) 15 mg/kg via the tail vein in anesthetized animals. Hydromorphone 120 μg was intraperitoneally injected at 15 min before LPS injection in WT+ ALI+ HM group and Nrf2 KO+ ALI+ HM group, and the equal volume of normal saline was given instead in control groups. The animals were sacrificed after anesthesia at 12 h after LPS injection, and lung tissues were obtained for examination of the pathological changes which were scored and Golgi ultrastructure (with a transmission electron microscope) and for determination of the content of malondialdehyde (MDA), activity of superoxide dismutase (SOD), and expression of Nrf2, HO-1 and Golgi stress-related markers (Golgi matrix protein 130 [GM130], Golgi autoantigen 97 kDa [Golgin-97], ATPase secretory pathway Ca 2+ Transporting 1 [ATP2C1], Golgi phosphoprotein 3 [GOLPH3]) (by Western blot). Results:Compared with WT+ Con group and Nrf2 KO+ Con group, the lung injury scores and content of MDA were significantly increased, the activity of SOD was decreased, the expression of GM130, Golgin-97 and ATP2C1 was down-regulated, the expression of GOLPH3 was up-regulated ( P<0.05), no significant changes were found in the expression of Nrf2 and HO-1 ( P>0.05), and the damage to the Golgi apparatus was aggravated in WT+ ALI group and Nrf2 KO+ ALI group. Compared with WT+ ALI group, the lung injury scores and content of MDA were significantly decreased, the activity of SOD was increased, the expression of Nrf2, HO-1, GM130, Golgin-97 and ATP2C1 was up-regulated, the expression of GOLPH3 was down-regulated ( P<0.05), and the damage to the Golgi apparatus was significantly attenuated in WT+ ALI+ HM group. Compared with Nrf2 KO+ ALI group, the lung injury scores were significantly decreased, and the activity of SOD was increased ( P<0.05), no significant changes were found in the content of MDA and expression of Nrf2, HO-1, GM130, Golgin-97, ATP2C1 and GOLPH3 ( P>0.05), and no significant reduction in the damage to the Golgi apparatus was found in Nrf2 KO+ ALI+ HM group. Compared with WT+ ALI+ HM group, the lung injury scores and content of MDA were significantly increased, the activity of SOD was decreased, the expression of Nrf2, HO-1, GM130, Golgin-97 and ATP2C1 was down-regulated, the expression of GOLPH3 was up-regulated ( P<0.05), and the damage to the Golgi apparatus was aggravated in Nrf2 KO+ ALI+ HM group. Conclusions:Nrf2/HO-1 signaling pathway is involved in the attenuation of endotoxin-induced ALI by hydromorphone, and it is associated with the inhibition of Golgi stress.

The enterotoxigenic Escherichia coli (ETEC) infection is a major factor restricting the development of animal husbandry. However, the abuse of antibiotics will lead to the antibiotic residues and emergence of antibiotic-resistant bacteria. The existing vaccines face challenges in stimulating intestinal immunity, demonstrating limited prevention effects. Therefore, it is indispensable to develop a new vaccine that is safe and suitable as a feed additive to activate intestinal immunity. This study constructed yeast-cell microcapsules (YCM) carrying the DNA vaccine against ETEC by genetic engineering. Furthermore, animal experiments were carried out to explore the regulatory effects of feeding YCM on the intestinal immune system and intestinal microbiota. Saccharomyces cerevisiae was selected as the oral delivery vehicle (microcapsules) of the DNA vaccine. The codon-optimized nucleic acid sequence of K88, the main antigen of mammal-derived ETEC, was synthesized, and the yeast shuttle vector containing the corresponding DNA vaccine expression cassette was constructed by DNA recombination. The recombinant strain of YCM was prepared by transforming JMY1. Additionally, the characteristics of the YCM strain and its feasibility as an oral vaccine were comprehensively evaluated by the fluorescence reporter assay, gastrointestinal fluid tolerance assay, intestinal epithelial cell adhesion assay, intestinal retention assessment, antiserum detection, and intestinal microbiota detection. The experimental results showed that the DNA vaccine expression cassette was expressed in mammals, and the recombinant strain of YCM could tolerate up to 8 hours of gastrointestinal fluid digestion and had good adhesion to intestinal epithelial cells. The results of mouse feeding experiments indicated that the recombinant strain of YCM could stay in the intestinal tract for at least two weeks, and the DNA vaccine expression cassette carried by YCM entered the intestinal immune system and triggered an immune response to induce the production of specific antibodies. Moreover, feeding YCM recombinant bacteria also improved the abundance of gut microbiota in mice, demonstrating a positive effect in regulating intestinal flora. In summary, we prepared the recombinant strain of YCM carrying the DNA vaccine against ETEC and comprehensively evaluated its characteristics and feasibility as an oral vaccine. Feeding the recombinant YCM could induce specific immune responses and regulate intestinal microbiota. The findings provide a reference for the immunoprevention of ETEC-related animal diseases.
Animals ; Enterotoxigenic Escherichia coli/genetics* ; Saccharomyces cerevisiae/metabolism* ; Vaccines, DNA/genetics* ; Mice ; Escherichia coli Infections/immunology* ; Escherichia coli Vaccines/genetics* ; Capsules ; Mice, Inbred BALB C ; Female

Objective:To evaluate the clinical efficacy of TCM acupoint application combined with Fuyang cupping in the treatment of chronic bronchitis with lung and kidney yin deficiency syndrome.Methods:A randomized controlled trial study was conducted. From July 2022 to July 2024, 96 patients with chronic bronchitis in our hospital were selected as the observation subjects and divided into two groups using a random number table method, with 48 cases in each group. The control group received treatment with Fuyang cupping, while the study group received treatment with Fuyang cupping and acupoint application. Both groups were treated continuously for 14 days. TCM syndrome scores were performed before and after treatment, and serum TNF-α, CRP, and procalcitonin (PCT) levels were detected using ELISA. Arterial oxygen partial pressure (PaO 2) and carbon dioxide partial pressure (PaCO 2) were measured using a blood gas analyzer. FVC, FEV1, and FEV1 percentage of predicted values (FEV1% predicted value) and maximum expiratory flow (PEF) were measured using a pulmonary function analyzer. The relief time of clinical symptoms such as cough, sputum production, and wheezing during the treatment period were observed and recorded, and the clinical efficacy was evaluated. Results:The total effective rate of the research group was 95.83% (46/48), while that of the control group was 79.16% (38/48), with statistical significance ( χ2=6.10, P=0.013). After treatment, the scores of cough, sputum, wheezing, wheezing, and dry throat and mouth in the study group were lower than those in the control group ( t values were 4.86, 5.89, 5.23, 4.10, 5.49, respectively, P<0.01); PaO 2 level [(95.63 ± 6.45) mmHg vs. (88.44 ± 5.76) mmHg, t=5.76] was higher than of the control group ( P<0.01), and the PaCO 2 level [(40.12 ± 4.28) mmHg vs. (45.63 ± 4.36) mmHg, t=6.25] was lower than of the control group ( P<0.01); the levels of FEV1 [(2.64 ± 0.69) L vs. (1.97 ± 0.74) L, t=4.59], FEV1% predicted value (79.25 ± 11.27 vs. 62.55 ± 10.56, t=7.49), FVC [(3.05 ± 1.04) L vs. (2.06 ± 0.96) L, t=4.85], and PEF [(241.54 ± 19.85) L/min vs. (221.36 ± 19.65) L/min, t=5.01] were higher than those in the control group ( P<0.01); the levels of TNF-α, CRP and PCT in the serum were lower than those in the control group ( t values were 4.39, 6.51 and 18.59, respectively, P<0.001). Conclusion:TCM acupoint application combined with Fuyang cupping can effectively improve the TCM syndrome score and lung function, inhibit inflammatory factor levels, and improve clinical efficacy in chronic bronchitis patients with lung and kidney yin deficiency syndrome.

Objective:To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase (HO-1) signaling pathway in the attenuation of endotoxin-induced acute lung injury (ALI) by hydromorphone and the relationship with Golgi apparatus stress (GA stress) in mice.Methods:Eighteen SPF wild-type (WT) and 18 Nrf2 knockout (Nrf2 KO) male C57BL/6J mice, aged 6-8 weeks, weighing 18-20 g, were divided into 3 groups ( n=6 each) using a random number table method: control groups (WT+ Con group, Nrf2 KO+ Con group), ALI groups (WT+ ALI group, Nrf2 KO+ ALI group) and ALI+ hydromorphone groups (WT+ ALI+ HM group, Nrf2 KO+ ALI+ HM group). ALI was induced by injecting lipopolysaccharide (LPS) 15 mg/kg via the tail vein in anesthetized animals. Hydromorphone 120 μg was intraperitoneally injected at 15 min before LPS injection in WT+ ALI+ HM group and Nrf2 KO+ ALI+ HM group, and the equal volume of normal saline was given instead in control groups. The animals were sacrificed after anesthesia at 12 h after LPS injection, and lung tissues were obtained for examination of the pathological changes which were scored and Golgi ultrastructure (with a transmission electron microscope) and for determination of the content of malondialdehyde (MDA), activity of superoxide dismutase (SOD), and expression of Nrf2, HO-1 and Golgi stress-related markers (Golgi matrix protein 130 [GM130], Golgi autoantigen 97 kDa [Golgin-97], ATPase secretory pathway Ca 2+ Transporting 1 [ATP2C1], Golgi phosphoprotein 3 [GOLPH3]) (by Western blot). Results:Compared with WT+ Con group and Nrf2 KO+ Con group, the lung injury scores and content of MDA were significantly increased, the activity of SOD was decreased, the expression of GM130, Golgin-97 and ATP2C1 was down-regulated, the expression of GOLPH3 was up-regulated ( P<0.05), no significant changes were found in the expression of Nrf2 and HO-1 ( P>0.05), and the damage to the Golgi apparatus was aggravated in WT+ ALI group and Nrf2 KO+ ALI group. Compared with WT+ ALI group, the lung injury scores and content of MDA were significantly decreased, the activity of SOD was increased, the expression of Nrf2, HO-1, GM130, Golgin-97 and ATP2C1 was up-regulated, the expression of GOLPH3 was down-regulated ( P<0.05), and the damage to the Golgi apparatus was significantly attenuated in WT+ ALI+ HM group. Compared with Nrf2 KO+ ALI group, the lung injury scores were significantly decreased, and the activity of SOD was increased ( P<0.05), no significant changes were found in the content of MDA and expression of Nrf2, HO-1, GM130, Golgin-97, ATP2C1 and GOLPH3 ( P>0.05), and no significant reduction in the damage to the Golgi apparatus was found in Nrf2 KO+ ALI+ HM group. Compared with WT+ ALI+ HM group, the lung injury scores and content of MDA were significantly increased, the activity of SOD was decreased, the expression of Nrf2, HO-1, GM130, Golgin-97 and ATP2C1 was down-regulated, the expression of GOLPH3 was up-regulated ( P<0.05), and the damage to the Golgi apparatus was aggravated in Nrf2 KO+ ALI+ HM group. Conclusions:Nrf2/HO-1 signaling pathway is involved in the attenuation of endotoxin-induced ALI by hydromorphone, and it is associated with the inhibition of Golgi stress.

Objective To investigate and observe the influence degree of continue nursing for the self -effi-cacy and compliance of patients with coronary heart disease.Methods 90 patients with coronary heart disease were selected as the research subjects,and 90 cases were divided into control group(conventional nursing of coronary heart disease group) 45 cases and observation group(continue nursing group) 45 cases by the method of random number table.Then,the self -efficacy,treatment compliance and satisfaction degree for the nursing of the two groups before the nursing and at first,second and fourth week after the nursing were compared.Results The self -efficacy and treatment compliance of the two groups before the nursing had no significant differences (all P >0.05),while the very high rates of the observation group's self -efficacy at first,second and fourth week after the nursing were 40.00%, 57.78% and 77.78% respectively,the higher rates of treatment compliance were 64.44%,84.44% and 97.78%respectively,the total satisfaction rates for the nursing were 97.78%,100.00%,100.00% respectively,which were all higher than those of the control group(all P <0.05),the differences were significant.Conclusion The influence of continue nursing for the self -efficacy and compliance of patients with coronary heart disease are more active,and it is more definitely to the patients,so the nursing mode is more suitable to patients with coronary heart disease.