OBJECTIVE: To investigate whether auraptene (AUR) exerts a protective effect on acute diquat (DQ)-induced liver injury in mice and explore its underlying mechanisms. METHODS: Forty SPF-grade healthy male C57BL/6 mice were randomly divided into normal control group (Control group), DQ poisoning model group (DQ group), AUR treatment group (DQ+AUR group), and AUR control group (AUR group), with 10 mice in each group. The DQ poisoning model was established via a single intraperitoneal injection of 40 mg/kg DQ aqueous solution (0.5 mL); Control group and AUR group received an equal volume of pure water intraperitoneally. Four hours post-modeling, DQ+AUR group and AUR group were administered 0.5 mg/kg AUR aqueous solution (0.2 mL) by gavage once daily for 7 consecutive days, while Control group and DQ group received pure water. Blood and liver tissues were collected after anesthesia on day 7. Liver ultrastructure was observed by transmission electron microscopy. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured via enzyme-linked immunosorbent assay (ELISA). Hepatic glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA) levels were detected using WST-1, thiobarbituric acid (TBA), and enzymatic reaction methods, respectively. Protein expression of nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), Kelch-like ECH-associated protein 1 (Keap1), and activated caspase-9 in liver tissues was analyzed by Western blotting. RESULTS: Transmission electron microscopy revealed that mitochondria in the Control group exhibited mild swelling, uneven distribution of matrix, and a small number of cristae fractures. In the AUR group, mitochondria showed mild swelling, with no obvious disruption of cristae structure. In the DQ group, mitochondria demonstrated marked swelling and increased volume, matrix dissolution, loss and fragmentation of cristae, and extensive vacuolization. In contrast, the DQ+AUR group showed significantly reduced mitochondrial swelling, volume increase, matrix dissolution, cristae loss and fragmentation, and vacuolization compared to the DQ group. Compared with the DQ group, the DQ+AUR group exhibited significantly lower serum AST levels (U/L: 173.45±23.60 vs. 255.33±41.51), ALT levels (U/L: 51.77±21.63 vs. 100.70±32.35), and hepatic MDA levels (μmol/g: 12.40±2.76 vs. 19.74±4.10), along with higher hepatic GSH levels (mmol/g: 37.65±14.95 vs. 20.58±8.52) and SOD levels (kU/g: 124.10±33.77 vs. 82.81±22.00), the differences were statistically significant (all P < 0.05). Western blotting showed upregulated Nrf2 expression (Nrf2/β-actin: 0.87±0.37 vs. 0.53±0.22) and HO-1 expression (HO-1/β-actin: 1.06±0.22 vs. 0.49±0.08), and downregulated Keap1 expression (Keap1/β-actin: 0.82±0.12 vs. 1.52±0.76) and activated caspase-9 expression (activated caspase-9/β-actin: 1.16±0.28 vs. 1.71±0.30) in the DQ+AUR group compared to the DQ group (all P < 0.05). CONCLUSION AUR attenuates DQ-induced acute liver injury in mice by activating the Keap1/Nrf2 signaling pathway.
Animals ; Male ; Mice ; Mice, Inbred C57BL ; Liver/pathology* ; Chemical and Drug Induced Liver Injury/drug therapy* ; Diquat/poisoning* ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; Apoptosis ; Coumarins

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective:To investigate the protective effect of quercetin (QR) on acute liver injury induced by diquat (DQ) poisoning in mice and its mechanism.Methods:Eighty healthy male C57BL/6 mice with SPF grade were randomly divided into control group, DQ model group, QR treatment group, and QR control group, with 20 mice in each group. The DQ poisoning model was established by a one-time intraperitoneal injection of DQ solution (40 mg/kg); the control and QR control groups received equivalent amounts of distilled water through intraperitoneal injection. Four hours after modeling, the QR treatment group and the QR control group received 0.5 mL QR solution (50 mg/kg) through gavage. Meanwhile, an equivalent amount of distilled water was given orally to the control group and the DQ model group. The treatments above were administered once daily for seven consecutive days. Afterwards, the mice were anesthetized, blood and liver tissues were collected for following tests: changes in the structure of mice liver tissue were observed using transmission electron microscopy; the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using enzyme linked immunosorbent assay (ELISA); the levels of glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA) in liver tissues were measured using the water-soluble tetrazolium-1 (WST-1) method, the thiobarbituric acid (TBA) method, and enzymatic methods, respectively; the protein expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), Kelch-like ECH-associated protein 1 (Keap1), and activated caspase-9 in liver tissues were detected using Western blotting.Results:Severe mitochondrial damage was observed in the liver tissues of mice in the DQ model group using transmission electron microscopy, yet mitochondrial damage in the QR treatment group showed significant alleviation. Compared to the control group, the DQ model group had significantly increased levels of MDA in liver tissue, serum AST, and ALT, yet had significantly decreased levels of GSH and SOD in liver tissue. In comparison to the DQ model group, the QR treatment group exhibited significant reductions in serum levels of ALT and AST, as well as MDA levels in liver tissue [ALT (U/L): 52.60±6.44 vs. 95.70±8.00, AST (U/L): 170.45±19.33 vs. 251.10±13.09, MDA (nmol/mg): 12.63±3.41 vs. 18.04±3.72], and notable increases in GSH and SOD levels in liver tissue [GSH (μmol/mg): 39.49±6.33 vs. 20.26±3.96, SOD (U/mg): 121.40±11.75 vs. 81.67±10.01], all the differences were statistically significant (all P < 0.01). Western blotting results indicated that the protein expressions of Nrf2 and HO-1 in liver tissues of the DQ model group were significantly decreased compared to the control group. On the other hand, the protein expressions of Keap1 and activated caspase-9 were conspicuously higher when compared to the control group. In comparison to the DQ model group, the QR treatment group showed a significant increase in the protein expressions of Nrf2 and HO-1 in liver tissues (Nrf2/β-actin: 1.17±0.08 vs. 0.92±0.45, HO-1/β-actin: 1.53±0.17 vs. 0.84±0.09). By contrast, there was a notable decrease in the protein expressions of Keap1 and activated caspase-9 (Keap1/β-actin: 0.48±0.06 vs. 1.22±0.09, activated caspase-9/β-actin: 1.17±0.12 vs. 1.59±0.30), the differences were statistically significant (all P < 0.01). Conclusion:QR may reduce acute liver injury induced by DQ poisoning in mice via activating Keap1/Nrf2 signaling pathway.

Objective:To investigate the effect of aucubin on behaviors and excessive activation of astrocytic in attention deficit/hyperactivity disorder (ADHD) model mice.Methods:Twelve wild-type C57BL/6 pregnant mice (female, clean grade) were intraperitoneally administered with esketamine (15 mg/kg) to establish an ADHD model in offspring mice. The offspring mice were divided into control+ saline group, control+ aucubin group, Ketamine+ saline group and Ketamine+ aucubin group according to the nest matching principle with 15 in each group.At 14 days after birth, mice in the control+ aucubin group and Ketamine+ aucubin group were administered with aucubin (5 mg/kg, once a day) by gavage for 5 days. Mice in control+ saline group and Ketamine+ saline group were administered with equal volume of 0.9% sodium chloride solution. The offspring mice were housed with their mothers in the same cage until 21 days after birth. Twenty-one days after birth, the offspring mice were evaluated by open field test and elevated plus maze tests. Immunofluorescence assay was used to detect the expression of glutamate decarboxylase 2 (GAD2), γ- aminobutyric acid (GABA) and glial fibrillary acidic protein (GFAP) in the amygdala. The morphological changes of astrocytes were quantitatively analyzed by Sholl analysis. GraphPad Prim 9.0.1 software was used for statistical analysis. The comparison of multiple groups was conducted by one-way ANOVA or Kruskal-Wallis test.Results:(1)The results of behavioral experiments showed that the total distance traveled in the open field test and the residence time in open arm of the elevated plus maze were statistically significant ( F=236.90, H=39.92, both P<0.001). The total distance ((7 044±249)mm, (22 891±2 175)mm, P<0.05) and the residence time in open arm(12.69(9.86, 17.24)s, 2.72(0.57, 3.87)s, P<0.05) of mice in Ketamine+ saline group were both higher than those in control+ saline group.The total distance((22 891±2 175)mm, (8 252±839)mm, P<0.05) and the the residence time in open arm(5.45(1.13, 10.99)s, 12.69(9.86, 17.24)s, P<0.05) of Ketamine+ aucubin group were both lower than those of Ketamine+ saline group.(2)The immunofluorescence results showed that the levels of GAD2, GABA and GFAP intensity in amygdala of mice in the four groups were statistically significant ( F=145.50, 50.08, 53.83, all P<0.05). Compared with control+ saline group, the fluorescence intensities of GAD2 ((100.00±9.60)%, (24.86±4.14)%, P<0.05) and GABA ((100.00±16.84))%, (25.48±5.70)%, P<0.05) of Ketamine+ saline group were down-regulated, and the GFAP((100.00±18.02)%, (223.80±25.85)%, P<0.05) was up-regulated. Compared with Ketamine+ saline group, the fluorescence intensities of GAD2 ((24.86±4.14)%, (56.08±6.55)%, P<0.05) and GABA((25.48±5.70)%, (52.59±15.74)%, P<0.05) in Ketamine+ aucubin group were up-regulated, but the fluorescence intensity of GFAP ((223.80±25.85)%, (157.10±22.10)%, P<0.05) was down-regulated.(3)Sholl analysis indicated that the number of the intersections between the astrocyte processes or the branches of astrocyte processes was statistically significant in the 4 groups ( F=12.47, P<0.05). Compared with control+ saline group, the number of the intersections in Ketamine+ saline group((2.07±0.48), (1.67±0.72), P<0.05) increased. While the number of the intersections in Ketamine+ aucubin group was lower than that of Ketamine+ saline group ((1.20±0.78), (2.07±0.48), P<0.05). Conclusion:Aucubin administration can alleviate ADHD-like behaviors in offspring mice, and the mechanism may be associated with the inhibition of excessive astrocytic activation.

Objective:To observe and evaluate the clinical effect of hyperbaric oxygen combined therapy on patients with non-arteritic anterior ischemic optic neuropathy (NAION).Methods:A total of 110 patients with NAION admitted to the Department of Ophthalmology of Zhengzhou Second Hospital from January 2017 to January 2019 who met the inclusion criteria were selected and divided into control group (55 cases) and hyperbaric oxygen group (55 cases) by random number table method. The control group was given conventional therapies such as hormone shock, microcirculation improvement, and nerve nutrition, while the hyperbaric oxygen group was given conventional treatment and hyperbaric oxygen therapy, with chamber pressure of 2.0 ATA, once a day, 10 times for a course of treatment for 30 days. The visual acuity, mean sensitivity (MS), mean defect (MD), pattern visual evoked potential (PVEP) amplitude, and latency period of the two groups were observed and analyzed to evaluate the efficacy of treatment. The effect of hyperbaric oxygen combined therapy on serum human chondroglycoprotein-39 (chtinase-3-like-1 protein, YKL-40) in patients with NAION was also observed.Results:Before treatment, there were no statistically significant differences in visual acuity, mean sensitivity, mean defect, PVEP amplitude, latency period, and serum YKL-40 between the two groups ( P>0.05). The visual MD in the hyperbaric oxygen group after treatment was lower than that in the control group after treatment and also lower than that before treatment in the same group, and the differences were statistically significant ( P<0.05). The visual acuity and MS in the hyperbaric oxygen group after treatment were higher than those in the control group and also those before treatment in the same group, and the differences were statistically significant ( P<0.05). The PVEP amplitudes of the patients in the hyperbaric oxygen group were higher than those in the control group and also those before treatment in the same group, and the latency period was earlier than that in the control group and also that before treatment in the same group, with statistically significant differences ( P<0.05). The level of serum YKL-40 reached the peak after 7 days of treatment. After 3, 7, 15 and 30 days of treatment, the levels of serum YKL-40 were all lower than those of the control group on the same day, with statistically significant differences ( P<0.05). After treatment, the total effective rate of the hyperbaric oxygen group (83.63%) was higher than that of the control group (74.55%), but there was no statistically significant difference ( P>0.05). Conclusion:Hyperbaric oxygen combined therapy can significantly improve the vision acuity, MS, MD, PVEP amplitude, and latency period of the patients with NAION, and reduce inflammatory response, which is beneficial to improve clinical efficacy.

Objective:To observe the clinical effect of the treatment combined with hyperbaric oxygen (HBO) on patients with herpes zoster.Methods:A total of 72 patients with herpes zoster treated in the Department of Dermatology of Zhengzhou Second Hospital from January 2018 to December 2019 were selected and divided into HBO group and control group by random number table method, with 36 patients in each group. The control group received acyclovir and other conventional treatments, while the HBO group received HBO on the basis of the treatments of the control group. The improvement of the skin lesions, scores of numeric rating scales (NRS) on pain, and efficacy of the two groups were observed and evaluated after treatment. After a 3-month follow-up, the occurrence of postherpetic neuralgia (PHN) was statistically analyzed.Results:The time of blistering relieving, incrustation, and complete incrustation of the HBO group were all lower than those of the control group with statistically significant difference ( P<0.05). The NRS scores of both groups were all lower than those before treatment with significantly statistical difference ( P<0.01); and the NRS scores of the HBO group were significantly lower than those of the control group after treatment ( P<0.05). The total effective rate of the HBO group (91.67%) was significantly higher than that of the control group (83.33%). The difference was statistically significant ( χ2=4.31, P=0.04). After treatment, 3 cases of PHN occurred in the HBO group, while 7 cases in the control group; the PHN incidence of the HBO group was lower than that of the control group with significantly statistical difference ( P<0.05). Conclusion:In treating herpes zoster, the treatment combined with HBO can achieve a satisfactory therapeutic effect, which is worthy of popularization and application in clinic.

Objective:To observe the clinical effect of the treatment combined with hyperbaric oxygen (HBO) on patients with herpes zoster.Methods:A total of 72 patients with herpes zoster treated in the Department of Dermatology of Zhengzhou Second Hospital from January 2018 to December 2019 were selected and divided into HBO group and control group by random number table method, with 36 patients in each group. The control group received acyclovir and other conventional treatments, while the HBO group received HBO on the basis of the treatments of the control group. The improvement of the skin lesions, scores of numeric rating scales (NRS) on pain, and efficacy of the two groups were observed and evaluated after treatment. After a 3-month follow-up, the occurrence of postherpetic neuralgia (PHN) was statistically analyzed.Results:The time of blistering relieving, incrustation, and complete incrustation of the HBO group were all lower than those of the control group with statistically significant difference ( P<0.05). The NRS scores of both groups were all lower than those before treatment with significantly statistical difference ( P<0.01); and the NRS scores of the HBO group were significantly lower than those of the control group after treatment ( P<0.05). The total effective rate of the HBO group (91.67%) was significantly higher than that of the control group (83.33%). The difference was statistically significant ( χ2=4.31, P=0.04). After treatment, 3 cases of PHN occurred in the HBO group, while 7 cases in the control group; the PHN incidence of the HBO group was lower than that of the control group with significantly statistical difference ( P<0.05). Conclusion:In treating herpes zoster, the treatment combined with HBO can achieve a satisfactory therapeutic effect, which is worthy of popularization and application in clinic.

Objective:To observe and evaluate the clinical effect of hyperbaric oxygen combined therapy on patients with non-arteritic anterior ischemic optic neuropathy (NAION).Methods:A total of 110 patients with NAION admitted to the Department of Ophthalmology of Zhengzhou Second Hospital from January 2017 to January 2019 who met the inclusion criteria were selected and divided into control group (55 cases) and hyperbaric oxygen group (55 cases) by random number table method. The control group was given conventional therapies such as hormone shock, microcirculation improvement, and nerve nutrition, while the hyperbaric oxygen group was given conventional treatment and hyperbaric oxygen therapy, with chamber pressure of 2.0 ATA, once a day, 10 times for a course of treatment for 30 days. The visual acuity, mean sensitivity (MS), mean defect (MD), pattern visual evoked potential (PVEP) amplitude, and latency period of the two groups were observed and analyzed to evaluate the efficacy of treatment. The effect of hyperbaric oxygen combined therapy on serum human chondroglycoprotein-39 (chtinase-3-like-1 protein, YKL-40) in patients with NAION was also observed.Results:Before treatment, there were no statistically significant differences in visual acuity, mean sensitivity, mean defect, PVEP amplitude, latency period, and serum YKL-40 between the two groups ( P>0.05). The visual MD in the hyperbaric oxygen group after treatment was lower than that in the control group after treatment and also lower than that before treatment in the same group, and the differences were statistically significant ( P<0.05). The visual acuity and MS in the hyperbaric oxygen group after treatment were higher than those in the control group and also those before treatment in the same group, and the differences were statistically significant ( P<0.05). The PVEP amplitudes of the patients in the hyperbaric oxygen group were higher than those in the control group and also those before treatment in the same group, and the latency period was earlier than that in the control group and also that before treatment in the same group, with statistically significant differences ( P<0.05). The level of serum YKL-40 reached the peak after 7 days of treatment. After 3, 7, 15 and 30 days of treatment, the levels of serum YKL-40 were all lower than those of the control group on the same day, with statistically significant differences ( P<0.05). After treatment, the total effective rate of the hyperbaric oxygen group (83.63%) was higher than that of the control group (74.55%), but there was no statistically significant difference ( P>0.05). Conclusion:Hyperbaric oxygen combined therapy can significantly improve the vision acuity, MS, MD, PVEP amplitude, and latency period of the patients with NAION, and reduce inflammatory response, which is beneficial to improve clinical efficacy.

Objective: To observe and explore the therapeutic efficacy of hyperbaric oxygen (HBO) in the treatment of non-arteritic anterior ischemic optic neuropathy (NAION). Methods: A total of 139 NAION patients were randomly divided into a control group of 72 and a hyperbaric oxygen group of 67. Both groups were given conventional drugs including prednisolone, mecobalamin and compound anisodine, while the hyperbaric oxygen group was additionally provided with hyperbaric oxygen treatment at a pressure of 0.2MPa once a day for 30 days. Each day′s treatment lasted for 110 minutes, including 20 minutes at increasing pressure, 20 minutes decreasing and 60 minutes with the pressure stable at 0.2MPa. Before and after the 30-day treatment, the visual acuity and visual mean sensitivity (MS) of the two groups were observed and compared. Results: There was no significant difference between the control group and the hyperbaric oxygen group in terms of average visual acuity or visual MS before the treatment. Afterward the average visual acuity (4.88±0.25) and visual MS (16.68±1.19) of the hyperbaric oxygen group were significantly higher than before the treatment and significantly better than those of the control group. The total effective rate of the hyperbaric oxygen group was 91%, significantly higher than that of the control group (75%). Conclusions Conventional treatment combined with hyperbaric oxygen therapy can significantly promote the visual acuity and visual MS of NAION patients.