Erratum - LncRNA gadd7 promotes mitochondrial membrane potential decrease and apoptosis of alveolar type II epithelial cells by positively regulating MFN1 in an in vitro model of hyperoxia-induced acute lung injury.

This corrects the article published in European Journal of Histochemistry 2023;67:3535. doi: 10.4081/ejh.2023.3535.

[Design and application of a kind of anti-bedsore turning pad].

Patients who are bedridden are for a long-time prone to develop bedsores, especially in the hip and sacral areas and limbs, which causes eczema, ulcers, infection and other complications, resulting in pain and more medical costs. Therefore, the medical staff of the Second Affiliated Hospital of Zunyi Medical University designed and developed a kind of anti-bedsore turning pad, and has obtained the national utility model patent (ZL 2021 2 3004923.9), which is suitable for various long-term bedridden patients. The anti-bedsore turning pad includes the center axis of the turning pad, and ventilation pad 1 and ventilation pad 2 designed on the left and right of turning pad center axis. Under the ventilation pad 1 and the ventilation pad 2, the air pad 1 and the air pad 2 are respectively designed. There is a bedspread connected with ventilation pad 1 and ventilation pad 2 on the inflatable pad 1 and the inflatable pad 2. Through the design of inflatable pad 1 and inflatable pad 2, the left and right of the anti-bedsore turning pad can be lowered or raised independently, which is convenient for the patient's body to tilt and turn over, and can significantly reduce the number of nursing staff and the burden of nursing staff when turning over. In addition, it is convenient to replace the force site at any time and reduce the occurrence of pressure ulcers caused by long-term pressure on the force site. Through the design of ventilation cushion 1 and ventilation cushion 2, the internal gas flow of the turning pad can be made, and the ventilation between the patient and the turning pad can be kept dry, so as to reduce the occurrence of eczema, ulcers or infection and other complications, and ultimately reduce the occurrence of bedsores. In addition, through the design of the most superficial limb pad, the patient's limb can be appropriately elevated or massaged, which increases the comfort of the patient. The anti-bedsore turning pad is simple and effective, and can be widely used in long-term bedridden patients.

LncRNA gadd7 promotes mitochondrial membrane potential decrease and apoptosis of alveolar type II epithelial cells by positively regulating MFN1 in an in vitro model of hyperoxia-induced acute lung injury.

The mortality and morbidity rates of ovarian cancer (OC) are high, but the underlying mechanisms of OC have not been characterized. In this study, we determined the role of Rho GTPase Activating Protein 30 (ARHGAP30) in OC progression. We measured ARHGAP30 abundance in OC tissue samples and cells using immunohistochemistry (IHC) and RT-qPCR. EdU, transwell, and annexin V/PI apoptosis assays were used to evaluate proliferation, invasiveness, and apoptosis of OC cells, respectively. The results showed that ARHGAP30 was overexpressed in OC tissue samples and cells. Inhibition of ARHGAP30 suppressed growth and metastasis of OC cells, and enhanced  apoptosis. Knockdown of ARHGAP30 in OC cells significantly inhibited the PI3K/AKT/mTOR pathway. Treatment with the PI3K/AKT/mTOR pathway inhibitor buparlisib simulated the effects of ARHGAP30 knockdown on growth, invasiveness, and apoptosis of OC cells. Following buparlisib treatment, the expression levels of p-PI3K, p-AKT, and p-mTOR were significantly decreased. Furthermore, buparlisib inhibited the effects of ARHGAP30 upregulation on OC cell growth and invasiveness. In conclusion, ARHGAP30 regulated the PI3K/AKT/mTOR pathway to promote progression of OC.

USP10 alleviates sepsis-induced acute kidney injury by regulating Sirt6-mediated Nrf2/ARE signaling pathway.

BACKGROUND: Severe sepsis, a major health problem worldwide, has become one of the leading causes of death in ICU patients. Further study on the pathogenesis and treatment of acute kidney injury (AKI) is of great significance to reduce high mortality rate of sepsis. In this study, the mechanism by which ubiquitin specific peptidase 10 (USP10) reduces sepsis-induced AKI was investigated. Ligation and perforation of cecum (CLP) was employed to establish C57BL/6 mouse models of sepsis. Hematoxylin-eosin (H&E) staining was performed to detect renal injury. The concentrations of serum creatinine (Cr), urea nitrogen (BUN) and cystatin C (Cys C) were determined using a QuantiChrom™ Urea Assay kit. RT-qPCR and western blot were conducted to assess the USP10 expression level. DHE staining was used to detect reactive oxygen species (ROS) levels. H2O2, MDA and SOD levels were assessed using corresponding colorimetric kits. Western blot was used to examine the expression levels of Bcl-2, Bax, cleaved caspase-3, Sirt6, Nrf2 and HO-1. MTT assay was used to determine cell viability, whereas TUNEL staining and flow cytometry were used to assess cell apoptosis. RESULTS: In this study, we found that USP10 was decreased in CLP-induced mouse renal tissues. We identified that USP10 alleviated renal dysfunction induced by CLP. Moreover, USP10 was found to reduce oxidative stress, and abated LPS-induced renal tubular epithelial cell injury and apoptosis. Finally, we discovered that USP10 promoted activation of the NRF2/HO-1 pathway through SIRT6 and attenuated LPS-induced renal tubular epithelial cell injury. CONCLUSIONS: This study found that USP10 activates the NRF2/ARE signaling through SIRT6. USP10 alleviates sepsis-induced renal dysfunction and reduces renal tubular epithelial cell apoptosis and oxidative stress.

Dexmedetomidine versus fentanyl as adjuvants to ropivacaine for epidural anaesthesia: A systematic review and meta-analysis.

BACKGROUND: Several randomized controlled trials (RCTs) have evaluated the use of dexmedetomidine versus fentanyl as adjuvants to ropivacaine for epidural anaesthesia, yet the results are conflicting. We aimed to conduct this meta-analysis to assess the effects and safety of dexmedetomidine and fentanyl as adjuvants to ropivacaine for epidural anaesthesia. METHODS: Cochrane Library et al databases were searched from inception to Jan 28, 2020. The synthesised effect sizes were presented as mean difference (MD) or odd of risk (OR) and 95% confidence intervals (95% CI). RESULTS: Nine RCTs with a total of 672 patients were included. The synthesised results indicated that the mean time to the onset of sensory block (MD: -2.82, 95% CI (-4.84, -0.80)), and the time to maximum motor block (MD: -4.35, 95% CI (-7.31, -1.40)) in dexmedetomidine group was significantly less than that of fentanyl group, while the mean time to rescue analgesia in dexmedetomidine group was significantly increased (MD: 99.13, 95% CI (82.89, 115.37)). The incidence of nausea and vomiting (OR: 0.43, 95% CI (0.29, 0.66)), and shivering (OR: 0.34, 95% CI (0.18, 0.63)) in dexmedetomidine group was significantly reduced, yet the incidence of oral dryness in dexmedetomidine group was significantly increased OR: 5.31, 95% CI (1.69, 16.69)). CONCLUSIONS: Dexmedetomidine is better than fentanyl as adjuvant to ropivacaine for epidural anaesthesia with better effects and less adverse events.

miR-21-5p Suppresses Mitophagy to Alleviate Hyperoxia-Induced Acute Lung Injury by Directly Targeting PGAM5.

Hyperoxia-induced acute lung injury (HALI) is a severe side effect of refractory hypoxemia treatment, for which no effective therapeutic strategy is available. Here, we found that the lung miR-21-5p level was significantly decreased in the rats subjected to hyperoxia. Further, we presented evidence that miR-21-5p was a crucial regulator of mitophagy and mitochondrial dysfunction. Moreover, it proved that miR-21-5p regulated hyperoxia-induced mitophagy and mitochondrial dysfunction by directly binding to the target gene PGAM5. In conclusion, for the first time, we found that miR-21-5p could directly suppress mitophagy and mitochondrial damage during HALI formation.

[Changes of small intestinal villi microcirculation in sidestream dark-field imaging with different target blood pressure in rabbits during endotoxin shock].

OBJECTIVE: Changes of small intestine villus microcirculation perfusion in sidestream dark-field (SDF) imaging in the rabbits during endotoxic shock after fluid resuscitation with different target mean arterial pressure (MAP), and evaluation of feasibility of monitoring small intestine villus microcirculation by SDF were studied. METHODS: Sixty standard New Zealand white rabbits were randomly divided into two groups: low target MAP group (group A, n = 30) and high target MAP group (group B, n = 30). Fistula operation of ileum was made in vitro, and lipopolysaccharide (LPS, 2 mg/kg) was injected to establish endotoxic shock model. Group A was administered with the lower dose fluid resuscitation (lactated Ringer solution, 20 mL×kg-1×h-1) for target MAP of 65 mmHg (1 mmHg = 0.133 kPa); group B was administered with the higher dose fluid resuscitation (lactated Ringer solution, 30 mL×kg-1×h-1) for MAP of 80 mmHg. Continuous norepinephrine intravenous injection (0.5-1.0 µg×kg-1×min-1) was administered only after fluid therapy couldn't reach the target MAP. The changes of small intestine villus microcirculation perfusion indexes such as vessels per villus (VV), proportion of perfused villi (PPV), microvascular flow index (MFI), borders of villus score (BVS), vessels villus score (VVS) were continuously observed and recorded before the shock, during the shock and after fluid resuscitation using SDF imaging. The differences of microcirculation perfusion were compared between two groups using the specific parameter evaluation system to determine severity of villi microcirculation and injury scores at different stages. RESULTS: VV and borders of villus were clear and contact before shock in two groups. After shock, VV, PPV were significantly decreased in both two groups, the borders of villus were destroyed, MFI, BVS, VVS and the total score of villi injury microcirculation were obviously and severely decreased. Partial blood flow of villous capillaries after fluid resuscitation was recovered in two groups, but the perfusion of some region was un-balanced with the outworn borders of villus. VV were rose as compared before and after fluid resuscitation in groups A and B (vessels: 1.21±0.22 vs. 0.81±0.12, 1.54±0.28 vs. 0.79±0.13), and PPV [(31±4)% vs. (12±2)%, (38±5)% vs. (13±3)%], MFI (1.55±0.09 vs. 1.09±0.03, 1.97±0.11 vs. 1.05±0.03), VVS (points: 1.22±0.08 vs. 0.89±0.02, 2.06±0.15 vs. 0.90±0.02) and the sum of MFI, BVS, VVS (3.70±0.19 vs. 2.85±0.07, 5.01±0.29 vs. 2.88±0.08) were significant rose (all P < 0.05). The recovery of group B was better than that of group A, and the injury score was reduced. But BVS were not increased in both groups compared with before and after shock (points: 0.93±0.05 vs. 0.87±0.03, 0.98±0.09 vs. 0.93±0.05, both P > 0.05). CONCLUSIONS: For the small intestine villus microcirculation perfusion, the higher target MAP (80 mmHg) after fluid resuscitation or/and vasoconstrictor drugs usage were probably better than the relatively lower target MAP (65 mmHg) during endotoxic shock. SDF imaging is a very promising technique for intestinal villi microcirculatory visualization and assessment.

[Screening type II alveolar epithelial cell apoptosis related microRNA].

OBJECTIVE: To screen type II alveolar epithelial cell (AECII) apoptosis related microRNA (miRNA) with gene chip technology, provide a new strategy for the prevention and treatment of hyperoxia-induced acute lung injury(HALI). METHODS: AECII of male Sprague-Dawley (SD) rats was primarily cultured for 36 hours, then exposed to 0.5 mmol/L H2O2 to establish apoptosis model. Transmission electron microscope (TEM) was used to identify AECII and to observe apoptosis cell morphology. Before and after H2O2 injury for 2.5, 6, 12 and 24 hours, fluorescence-activated cell sorting (FACS) was employed to detect apoptosis rate. Additionally, gene chip technology and real time polymerase chain reaction (RT-PCR) were used to screen and verify apoptosis related miRNA respectively. RESULTS: Microvilli and osmiophilic multilamellar body were found under TEM, which were the characteristic structure of AECII. This proved that the cells cultured were AECII. After H2O2 injury for 24 hours, cytoplasmic retraction, chromatin condensation and margination, microvilli and osmiophilic multilamellar body disappearance could be found under TEM. Compared with the blank control group, the apoptosis rate of AECII was significantly increased after exposed to 0.5 mmol/L H2O2, and gradually increased with time [the early apoptosis rate before and after H2O2 injury for 2.5, 6, 12 and 24 hours were (9.43±1.02)%, (18.38±2.91)%, (28.57±1.18)%, (35.83±2.66)% and (57.68±2.22)%, respectively, all P<0.05]. Compared with before H2O2 injury, cells at 24 hours accompanied a significantly changed miRNA expression profiling, in which apoptosis related miRNA had been screened, they were rno-miR-449a-5p, rno-miR-34b/c-5p, rno-miR-200a/c-3p, rno-miR-146a-5p, rno-miR-141-3p, rno-miR-21-5p, rno-miR-375-3p, rno-miR-29b-3p, rno-miR-214-5p, rno-miR-210-5p and rno-miR-214-3p,and the results of RT-PCR were consistent with the gene chip results. CONCLUSIONS: Screened out AECII apoptosis related miRNA, and miR-34 family may play a key role in AECII apoptosis regulation. Rno-miR-21-5p may be an important anti-apoptotic gene in AECII apoptosis regulation.

[The effect of heme oxygenase-1 on apoptosis and aquaporin-1 expression in type II primary alveolar epithelial cells in rat].

OBJECTIVE: To explore the effect of heme oxygenase-1 (HO-1) on apoptosis and expression of aquaporin-1 (AQP-1) in primary type II alveolar epithelial cells (AECII) in rats with hydrogen peroxide (H2O2) induced oxidative damage. METHODS: Lung tissue of male Sprague-Dawley (SD) rats was collected. Primary AECII were isolated, purified, and cultured for 24 hours, then they were divided into four groups: (1) normal group (treated with normal saline); (2) H2O2 injury group (treated with H2O2 0.5 mmol/L); (3) HO-1 control group (treated with HO-1 1 µmol/L); (4) HO-1 protection group (treated with HO-1 1 µmol/L and H2O2 0.5 mmol/L). Cells of each group were cultured for 12 hours after various treatment. The cell suspension was collected before and 1, 3, 6, 12 hours after treatment, the expression of AQP-1 was determined by Western blotting and the apoptosis rate was assessed with flow cytometer. RESULTS: The expression of AQP-1 in H2O2 injury group was significantly declined with time, and was lower than that in normal group at each time point after treatment. The expression of AQP-1 in HO-1 control group and HO-1 protection group was significantly increased with time, and was higher than that of other groups at each time point after treatment. The expression of AQP-1 in HO-1 protection group was significantly up-regulated compared with that in H2O2 injury group (1 hour: 60.81 ± 5.78 vs. 46.21 ± 4.81, 3 hours: 63.05 ± 9.61 vs. 39.32 ± 4.96, 6 hours: 92.59 ± 8.21 vs. 36.82 ± 4.32, 12 hours: 86.16 ± 14.84 vs. 34.88 ± 2.66, all P<0.05). No significant difference in apoptosis rate was found between normal group and HO-1 control group. The apoptosis rate in H2O2 injury group was increased with time, and was significantly higher than that of normal group at each time point. The apoptosis rate in HO-1 protection group was gradually increased within 3 hours after treatment, then decreased and remained stable after 6 hours, while it was significantly lower than that of H2O2 injury group at each time point [1 hour: (9.04 ± 2.17)% vs. (15.14 ± 2.47)%, 3 hours: (12.90 ± 2.04)% vs. (22.37 ± 4.84)%, 6 hours: (10.42 ± 1.68)% vs. (27.83 ± 3.93)%, 12 hours: (11.97 ± 1.91)% vs. (33.63 ± 6.61)%, all P<0.05]. A negative correlation was found between AQP-1 and apoptosis rate in H2O2 injury group (r=-0.723, P<0.001), and a regression correlation was found [y=672.548(0.914) (x), R²=0.597]. AQP-1 was not correlated with apoptosis rate in HO-1 protection group (r=0.210, P=0.193), but a regression correlation was found [y=e((3.130 - 59.654/x )), R²=0.225]. CONCLUSIONS: HO-1 could increase the expression of AQP-1 in H2O2 injured AECII of rat, and lower its apoptosis rate. Increase in the expression of AQP-1 may be the underlying mechanism of anti-oxygenation property of HO-1.

[The role of heme oxygenase 1 in hydrogen peroxide induced apoptosis and mitochondrial trans membrane potential change in rat primary type II alveolar epithelium cells].

OBJECTIVE: To understand the role of heme oxygenase-1 (HO-1) in hydrogen peroxide [H(2)O(2)] induced apoptosis and mitochondrial trans-membrane potential (MTMP) change in primary alveolar epithelial cell type II(AEC II). METHODS: Primary AEC II collected from healthy Sprague Dawley (SD) rats were cultured for 24 hours, then divided into four groups to be treated with: (1) saline; (2) H(2)O(2) (0.5 mmol/L); (3) H(2)O(2) +HO-1 (0.2 mmol/L); (4) H(2)O(2) +zinc original porphyrin IX (HO-1 inhibitor, 20 µmol/L). The morphology of cells in the cultures was examined by fluorescent microscopy 2.5 hours later, and the number of apoptotic cells / the MTMP determined by flow-cytometry 0.5, 1.0, 1.5, 2.0 and 2.5 hours later. RESULTS: Large number of cells in with green (early apoptotic) or red (later apoptotic) fluorescence were observed by microscope in cultures treated with H(2)O(2) , and H(2)O(2) + HO-1 inhibitor, but such cells were obviously fewer in HO-1 treated cultures. Compared with saline treated cells, H(2)O(2) treated cells had significantly higher apoptosis rate, that increased with time, reaching peak value 2.5 hours into the treatment [0.5 hour: (30.27 ± 0.74)% vs. (3.76 ± 0.81)%, 2.5 hours: (40.46 ± 0.91)% vs. (22.74 ± 0.60)%, both P < 0.05], while the rate of MTMP depolarization was significantly lower (0.99 ± 0.21 vs. 1.91 ± 0.16, P < 0.05) in these cells. Compared with H(2)O(2) treated cells, the apoptosis rate in HO-1 treated cells was significantly lower [0.5 hour: (5.99 ± 0.60)% vs. (30.27 ± 0.74)%, 2.5 hours: (22.69 ± 1.69)% vs. (40.46 ± 0.91)%, both P < 0.05], and their rate of MTMP depolarization higher (2.02 ± 0.12 vs. 0.99 ± 0.21, P < 0.05). Compared with HO-1 treated cells, HO-1 inhibitor treated cells had significantly higher apoptosis rate which reached peak value 2.5 hours into the treatment [0.5 hour: (30.73 ± 1.08)% vs. (5.99 ± 0.60)%, 2.5 hours: (41.38 ± 0.57)% vs. (22.69 ± 1.69)%, both P < 0.05], while rate of MTMP depolarization in these cells was significantly lower (0.98 ± 0.09 vs. 2.02 ± 0.12, P < 0.05). CONCLUSION: HO-1 could maintain the integrity of AEC II and stabilize their mitochondria membrane potential, protecting the cells from H(2)O(2) induced damage.

[The effect on anti oxidative damage with aspirin pretreatment in primary cultured rat type II alveolar epithelial cell].

OBJECTIVE: To investigate the protective effect of aspirin on primary cultured type II alveolar epithelial cell (AEC II), and the mechanism of its effect on anti oxidation damage. METHODS: The original generation of adult rat AEC II were cultured and purified. They were divided into normal saline (NS) group, hydrogen peroxide injury group (H(2)O(2) group), and 1, 2, 3 aspirin pretreatment groups (A1-3 groups).In H(2)O(2) group, 0.5 mmol/L H(2)O(2) was added to AEC II after 40 hours of culture to reproduce a cell oxidative injury model. In NS group, only NS was added to AEC II culture. To the A1-3 groups aspirin 50, 100 and 200 µmol/L were added respectively. Cell form, cell count and cell survival rate were observed at 3 hours after H(2)O(2) was given . Immunohistochemical and polymerase chain reaction (PCR) methods were used for the determination of heme oxygenase-1 (HO-1) protein and HO-1 mRNA (20, 40, 60 hours of culture). RESULTS: With trypsin digestion and immune adherence method AEC II could be harvested (2.0-2.5)× , and the purity and activity were both over 90%. Compared with NS group, gaps between cells were widened in H(2)O(2) group, cell account was reduced, and the survival rate (A value) was reduced significantly (0.054 6±0.004 0 vs. 0.103 8±0.009 9, P<0.01). Compared with H(2)O(2) group, in A1-3 groups the number of adherent cells was increased, cell morphology was intact, and no obvious cell shrinkage was found. Higher survival rate (A value) was found in A1-3 groups than that of H(2)O(2) group (0.066 9±0.003 9, 0.071 0±0.006 5, 0.078 7±0.009 2 vs. 0.054 6±0.004 0, all P<0.01). Compared with NS group, HO-1 protein and HO-1 mRNA expression in AEC II after 20, 40 and 60 hours of culture reached peak level at 60 hours, and they were increased significantly in A1-3 groups [protein (A value): 1.59±0.12, 1.60±0.09, 1.61±0.08 vs. 1.25±0.11; mRNA (the ratio of Ct value: 24.31±1.74, 30.45± 2.53, 32.63±3.74 vs. 22.99±1.95, all P<0.05]. There was no significant difference in HO-1 protein expression among A1-3 groups. CONCLUSION: There are significant protective effects of aspirin against anti oxidative damage in cultured AEC II cell. As expression of HO-1 is increased in aspirin groups, it may be considered as a protective factor against anti oxidative damage in AEC II cell culture.