Objective:To observe the effects of peptidylarginine deiminase 2 (PAD2) inhibitor AFM-30a on silicotic mice and its possible mechanisms.Methods:In May 2022, 40 SPF male C57BL/6J mice were randomly divided into control group, AFM-30a group, silicosis model group and AFM-30a treatment group, with 10 mice in each group. Silicosis model group and AFM-30a treatment group were perfused with silicon dioxide (SiO 2) suspension (10 mg/piece, 50 μl), and the other groups were perfused with an equal amount of sodium chloride solution. After 2 weeks, AFM-30a group and AFM-30a treatment group were intraperitoneally injected AFM-30a (20 mg/kg, 100 μl) daily, and mice of other groups were injected with equal amounts of sodium chloride solution for 4 weeks. Mouse RAW264.7 monocytes/macrophages were cultured in vitro and divided into blank control group, AFM-30a group (5 μmol/L), SiO 2 group (200 μg/ml), and SiO 2+AFM-30a group (200 μg/ml SiO 2 induction for 12 h, followed by 5 μmol/L AFM-30a treatment for 12 h). As well as blank control group, vimentin (Vim) group (2 μg/ml), citrullinated vimentin (Cit-Vim) group (2 μg/ml), and Cit-Vim+TLR4-C34 group (10 μmol/L TLR4-C34 treatment for 1 h, followed by 2 μg/ml Cit-Vim induction for 24 h). Hematoxylin Eosin (HE) and Masson staining were used to observe the pathological morphology of lung. The lung fieldclarity and lung texture of each group was observed by micro-CT. The number of positive cells was detected by tartrate resistant acid phosphatase (TRAP) staining. The localization and expression levels of PAD2, Cit-Vim, toll-like receptor 4 (TLR4) signaling and receptor activator of nuclear factor-κB ligand (RANKL) signaling proteins were measured by Immunofluorescence staining and Western blotting in vitro and in vivo. The experimental data were all presented as Mean±SD. A completely random design of one-way analysis of variance was used among the groups. The pduo comparison was performed using LSD test for homogeneity of variance and Tamhane's test for inconsistency. Results:Compared with the control group, the silicosis model group showed the formation of silicon nodules accompanied by collagen deposition, the silicosis model group showed thickened, and several high-density shadows of varying sizes in the lung field, and the number of TRAP positive cells in silicosis model group were increased significantly, the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signal-related proteins were also significantly increased in silicosis groupmodel ( P<0.05). Compared with the silicosis model group, the AFM-30a treatment group reduced deposition of collagen in lung, and the number of TRAP positive cells was decreased in AFM-30a treatment group. The expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins were significantly decreased in AFM-30a treatment group ( P<0.05). In vitro, compared with the blank control group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2 group were significantly increased ( P<0.05). Compared with the SiO 2 group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2+AFM-30a group were significantly decreased ( P<0.05). Compared with the blank control group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim group were significantly increased ( P<0.05). Compared with the Cit-Vim group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim+TLR4-C34 group were significantly decreased ( P<0.05) . Conclusion:PAD2 inhibitor AFM-30a may play an antagonisticrole in silicotic fibrosis in mice by potentialregulating TLR4 and RANKL signaling pathways.

Objective:To observe the effects of peptidylarginine deiminase 2 (PAD2) inhibitor AFM-30a on silicotic mice and its possible mechanisms.Methods:In May 2022, 40 SPF male C57BL/6J mice were randomly divided into control group, AFM-30a group, silicosis model group and AFM-30a treatment group, with 10 mice in each group. Silicosis model group and AFM-30a treatment group were perfused with silicon dioxide (SiO 2) suspension (10 mg/piece, 50 μl), and the other groups were perfused with an equal amount of sodium chloride solution. After 2 weeks, AFM-30a group and AFM-30a treatment group were intraperitoneally injected AFM-30a (20 mg/kg, 100 μl) daily, and mice of other groups were injected with equal amounts of sodium chloride solution for 4 weeks. Mouse RAW264.7 monocytes/macrophages were cultured in vitro and divided into blank control group, AFM-30a group (5 μmol/L), SiO 2 group (200 μg/ml), and SiO 2+AFM-30a group (200 μg/ml SiO 2 induction for 12 h, followed by 5 μmol/L AFM-30a treatment for 12 h). As well as blank control group, vimentin (Vim) group (2 μg/ml), citrullinated vimentin (Cit-Vim) group (2 μg/ml), and Cit-Vim+TLR4-C34 group (10 μmol/L TLR4-C34 treatment for 1 h, followed by 2 μg/ml Cit-Vim induction for 24 h). Hematoxylin Eosin (HE) and Masson staining were used to observe the pathological morphology of lung. The lung fieldclarity and lung texture of each group was observed by micro-CT. The number of positive cells was detected by tartrate resistant acid phosphatase (TRAP) staining. The localization and expression levels of PAD2, Cit-Vim, toll-like receptor 4 (TLR4) signaling and receptor activator of nuclear factor-κB ligand (RANKL) signaling proteins were measured by Immunofluorescence staining and Western blotting in vitro and in vivo. The experimental data were all presented as Mean±SD. A completely random design of one-way analysis of variance was used among the groups. The pduo comparison was performed using LSD test for homogeneity of variance and Tamhane's test for inconsistency. Results:Compared with the control group, the silicosis model group showed the formation of silicon nodules accompanied by collagen deposition, the silicosis model group showed thickened, and several high-density shadows of varying sizes in the lung field, and the number of TRAP positive cells in silicosis model group were increased significantly, the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signal-related proteins were also significantly increased in silicosis groupmodel ( P<0.05). Compared with the silicosis model group, the AFM-30a treatment group reduced deposition of collagen in lung, and the number of TRAP positive cells was decreased in AFM-30a treatment group. The expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins were significantly decreased in AFM-30a treatment group ( P<0.05). In vitro, compared with the blank control group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2 group were significantly increased ( P<0.05). Compared with the SiO 2 group, the number of TRAP positive cells and the expression levels of PAD2, Cit-Vim, TLR4 and RANKL signaling related proteins in the SiO 2+AFM-30a group were significantly decreased ( P<0.05). Compared with the blank control group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim group were significantly increased ( P<0.05). Compared with the Cit-Vim group, the expression levels of TLR4 and RANKL signaling related proteins in the Cit-Vim+TLR4-C34 group were significantly decreased ( P<0.05) . Conclusion:PAD2 inhibitor AFM-30a may play an antagonisticrole in silicotic fibrosis in mice by potentialregulating TLR4 and RANKL signaling pathways.

Background The senescence of alveolar type II epithelial cells is an important driving factor for the progression of silicotic fibrosis, and the regulatory effects of oxamate on the senescence of alveolar type II epithelial cells is still unclear. Objective To explore whether lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting senescence of alveolar type II epithelial cellsMethods This study was divided into two parts: in vivo experiments and in vitro experiments. In the first part, forty SPF C57BL/6J male mice were randomly divided into four groups with 10 in each group: control group, silicosis model group, low-dose oxamate treatment group, and high-dose oxamate treatment group. The silicotic mouse model was established by intratracheal instillation of 50 μL SiO₂ suspension (100 mg·mL⁻¹). The treatment models were prepared by intraperitoneal injection of 100 μL oxamate (225 mmol·L⁻¹ and 1125 mmol·L⁻¹). In the second part, induction of MLE-12 mouse alveolar type II epithelial cells was conducted with SiO₂. The in vitro experimental groups were ① SiO₂ induction groups: control group, 50 μg·mL⁻¹ SiO₂ group, 100 μg·mL⁻¹ SiO₂ group, and 200 μg·mL⁻¹ SiO₂ group, and ② oxamate treatment groups: control group, SiO₂ group (100 μg·mL⁻¹), low-dose oxamate (25 mmol·L⁻¹) treatment group, and high-dose oxamate (50 mmol·L⁻¹) treatment group. Pathological morphology of lung tissues was evaluated after hematoxylin-eosin (HE) staining; deposition of collagen in lung tissues was evaluated after sirius red staining; positive co-expression of prosurfactant protein C (Pro-SPC) and β-galactosidase was detected by immunofluorescence staining; positive expression of β-galactosidase in MLE-12 cells was detected by immunofluorescence staining. The protein expression levels of collagen type I (CoL I), fibronectin1 (FN1), hexokinase 2 (HK2), pyruvate kinase isozyme type M2 (PKM2), lactate dehydrogenase A (LDHA), p-ataxia telangiectasia and Rad3-related kinase (ATR), and cyclin-dependent kinase inhibitors p21, and p16 were detected by Western blotting. Results Compared with the control group, the protein expression levels of HK2, PKM2, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group and the SiO₂-induced MLE-12 cells (P＜0.05). The in vivo studies showed that, compared with the control group, the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the protein expression levels of CoL I, FN1, LDHA, p-ATR, p21, and p16 were significantly upregulated in the silicosis model group (P＜0.05). Compared with the silicosis model group, the oxamate treatment groups showed significant downregulation of the silicon nodule area, the collagen deposition area, the proportion of β-galactosidase positive cells, and the the CoL I, FN1, LDHA, p-ATR, p21, and p16 protein expression levels, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). The in vitro studies showed that, compared with the control group, the proportion of β-galactosidase positive cells and the protein expression levels of p-ATR, p21, and p16 were significantly upregulated in the SiO₂-induced group (P＜0.05). Compared with the SiO₂ group, the proportion of β-galactosidase positive cells and the LDHA, p-ATR, p21 and p16 protein expression levels were significantly downregulated in the oxamate treatment groups, and the high-dose oxamate treatment group showed a higher efficacy on these indicators than the low-dose oxamate treatment group (P＜0.05). Conclusion Lactate dehydrogenase inhibitor oxamate can alleviate silicotic fibrosis in mice by inhibiting the senescence of alveolar type II epithelial cells.

Background Pneumoconiosis is the most serious occupational disease in China, and silicosis accounts for about half of it. Any intervention effect of physical exercise as the key and core of lung rehabilitation training on silicosis is still unclear. Objective To explore potential intervention effect of physical exercise on silicotic mice. Methods Forty SPF C57BL/6 male mice were randomly divided into four groups, 10 in each group, including a control group, a physical exercise group, a silicosis model group, and a silicosis model + physical exercise intervention group. Silicotic mouse model was established by using 50 μL SiO₂ suspension (200 mg·mL⁻¹). A treadmill was used to prepare mice receiving physical exercise at 0° inclination, 12.3 m·min⁻¹, 60 min·d⁻¹, 5 d·week⁻¹ for 4 weeks. Pathological morphology of lung tissues was evaluated after hematoxylin-eosin (HE) staining; deposition of collagen in lung tissues was evaluated after Van Gieson (VG) staining; expression of p-protein kinase R-like endoplasmic reticulum kinase (PERK) was detected by immunofluorescence staining; expressions of cyclin dependent kinase inhibitors (p21) and p-p38 mitogen activated protein kinase (p38) were detected by immunohistochemistry. The protein expressions of endoplasmic reticulum stress signal factors [p-inositol-requiring enzyme-1α (p-IRE-1α), p-PERK, and p-eukaryotic initiation factor-2α (p-eIF-2α)], senescence signal factors (p-p53, p21, and p16), mitogen-activated protein kinase (MAPK) signal factors [p-p38, p-extracellular regulated protein kinases (p-ERK), and p-stress-activated protein kinase (p-JNK)] were detected by Western blotting. Results After designed acute SiO₂ exposure, the images of micro computed tomography (CT) showed high density shadows in lung tissues of the silicotic mice and less shadows in lung tissues of the physical exercise intervention mice. After HE staining, the proportions of silicotic nodule area in lung tissues was (18.67±3.89) % in the silicosis model group, and significantly decreased to (8.78±1.05) % in the silicosis model + physical exercise intervention group (P<0.05). After VG staining, the proportion of collagen fiber area of lung tissues was (10.37±2.18) % in the silicosis model group, and significantly decreased to (4.35±0.89) % in the silicosis model + physical exercise intervention group (P<0.05). The results of immunofluorescence staining showed that in the silicosis model group, the expression of p-PERK increased at the location of silicotic nodules, while in the silicotic model + physical exercise intervention group, the expression of p-PERK decreased. The immunohistochemical staining results showed that the expression of p21 and p-p38 increased in the lung tissues of the silicosis model group; the expression of p21 and p-p38 decreased in the lung tissues of the silicosis model + physical exercise intervention group. The results of Western blotting showed that compared with the control group, the expression levels of p-IRE-1α (0.11±0.03), p-PERK (0.95±0.40), p-eIF-2α (3.53±0.91), p-p53 (1.78±0.07), p21 (1.98±0.10), p16 (1.26±0.17), p-p38 (0.41±0.09), p-ERK (0.42±0.05), and p-JNK (3.20±1.23) of the silicosis model group were all upregulated (P<0.05). Compared with the silicosis model group, the expression levels of p-IRE-1α (0.03±0.01), p-PERK (0.31±0.12), p-eIF-2α (0.30±0.06), p-p53 (0.76±0.08), p21 (0.18±0.11), p16 (0.70±0.24), p-p38 (0.03±0.00), p-ERK (0.19±0.03), and p-JNK (0.46±0.21) of the silicosis model + physical exercise intervention group were downregulated (P<0.05). Conclusion Physical exercise may alleviate pulmonary fibrosis in silicotic mice, and inhibit abnormal expressions of endoplasmic reticulum stress signal, MAPK signal, and senescent signal.

Humans ; COVID-19 ; Quarantine ; Renal Dialysis ; SARS-CoV-2 ; Hospitals

Objective:To investigate the clinical efficacy and action mechanism of hyperbaric oxygen (HBO) combined with microsurgical lysis in the treatment of lipoma-type tethered cord syndrome (TCS).Methods:The clinical data of 59 patients with lipoma-type TCS treated in the Department of Orthopedics of Yantai Mountain Hospital and the Department of Orthopedics of Yantai Yuhuangding Hospital from January 2015 to January 2020 were retrospectively analyzed. According to the treatment, they were divided into observation group ( n=33) and control group ( n=26). The control group was treated with conventional microsurgical lysis, while the observation group was treated with HBO on the basis of the treatment in the control group. The clinical efficacy was evaluated according to the Kirollos grading system of postoperative TCS lysis. The optical densities ( D) of malondialdehyde (MDA) and superoxide dismutase (SOD) in cerebrospinal fluid were measured by spectrophotometer before microsurgical lysis and after 3 courses of treatment. Somatosensory evoked potential (SEP) was used to monitor and judge the efficacy of operation and the recovery of neurological function after treatment, including the amplitude, latency, and conduction velocity of posterior tibial nerve sensory evoked potential (PTNSEP). The reflexes, motor function, and the bladder and bowel function were evaluated according to the spina bifida neurological scale (SBNS). The bladder compliance, detrusor reflex, and detrusor strength after operation were measured by multifunctional urodynamic manometry. Postoperative complications, such as headache, nerve damage, cerebrospinal fluid leakage pseudocyst, and re-tethering were evaluated. Results:The total effective rate of the observation group (87.88%) was significantly higher than that of the control group (73.08%) ( χ2=4.991, P<0.05). There were significant differences in detrusor hyperreflexia, decreased bladder compliance, and detrusor weakness between the two groups after treatment ( F=6.534, P<0.05). After three weeks of treatment, compared with those before treatment, the levels of MDA in cerebrospinal fluid of the two groups were significantly decreased, and the levels of SOD of the two groups were significantly increased than those before treatment; the level of MDA in cerebrospinal fluid in the observation group was significantly lower than that of the control group, and the level of SOD in the observation group was significantly higher than that in the control group ( P<0.05). After 3 courses of treatment, compared with those before treatment, the conduction velocity and evoked potential amplitude of the two groups were significantly increased, and the latency was significantly shortened in the two groups; and the conduction velocity and evoked potential amplitude of the observation group were significantly increased than those in the control group, and the latency of the observation group was significantly lower than that of the control group ( P<0.05). In the six-month follow-up after operation, the total SBNS score of the observation group [(14.49±1.76) points] was significantly higher than that of the control group [(10.86±1.44) points], with a statistically significant difference ( χ2=5.782, P<0.05). The incidences of headache, cerebrospinal fluid leakage pseudocyst, nerve damage, and re-tethering in the observation groups were significantly lower than those in the control group ( F=6.395, P<0.05). Conclusion:HBO combined with microsurgical lysis is effective in the treatment of lipoma-type TCS, which can improve the anoxic-ischemic state of spinal cord neurons, accelerate the tibial nerve conduction velocity, significantly improve the comprehensive score of SBNS, and significantly reduce the incidence of postoperative complications. Thus, it is worthy of clinical application.

Objective:To investigate the clinical efficacy and action mechanism of hyperbaric oxygen (HBO) combined with microsurgical lysis in the treatment of lipoma-type tethered cord syndrome (TCS).Methods:The clinical data of 59 patients with lipoma-type TCS treated in the Department of Orthopedics of Yantai Mountain Hospital and the Department of Orthopedics of Yantai Yuhuangding Hospital from January 2015 to January 2020 were retrospectively analyzed. According to the treatment, they were divided into observation group ( n=33) and control group ( n=26). The control group was treated with conventional microsurgical lysis, while the observation group was treated with HBO on the basis of the treatment in the control group. The clinical efficacy was evaluated according to the Kirollos grading system of postoperative TCS lysis. The optical densities ( D) of malondialdehyde (MDA) and superoxide dismutase (SOD) in cerebrospinal fluid were measured by spectrophotometer before microsurgical lysis and after 3 courses of treatment. Somatosensory evoked potential (SEP) was used to monitor and judge the efficacy of operation and the recovery of neurological function after treatment, including the amplitude, latency, and conduction velocity of posterior tibial nerve sensory evoked potential (PTNSEP). The reflexes, motor function, and the bladder and bowel function were evaluated according to the spina bifida neurological scale (SBNS). The bladder compliance, detrusor reflex, and detrusor strength after operation were measured by multifunctional urodynamic manometry. Postoperative complications, such as headache, nerve damage, cerebrospinal fluid leakage pseudocyst, and re-tethering were evaluated. Results:The total effective rate of the observation group (87.88%) was significantly higher than that of the control group (73.08%) ( χ2=4.991, P<0.05). There were significant differences in detrusor hyperreflexia, decreased bladder compliance, and detrusor weakness between the two groups after treatment ( F=6.534, P<0.05). After three weeks of treatment, compared with those before treatment, the levels of MDA in cerebrospinal fluid of the two groups were significantly decreased, and the levels of SOD of the two groups were significantly increased than those before treatment; the level of MDA in cerebrospinal fluid in the observation group was significantly lower than that of the control group, and the level of SOD in the observation group was significantly higher than that in the control group ( P<0.05). After 3 courses of treatment, compared with those before treatment, the conduction velocity and evoked potential amplitude of the two groups were significantly increased, and the latency was significantly shortened in the two groups; and the conduction velocity and evoked potential amplitude of the observation group were significantly increased than those in the control group, and the latency of the observation group was significantly lower than that of the control group ( P<0.05). In the six-month follow-up after operation, the total SBNS score of the observation group [(14.49±1.76) points] was significantly higher than that of the control group [(10.86±1.44) points], with a statistically significant difference ( χ2=5.782, P<0.05). The incidences of headache, cerebrospinal fluid leakage pseudocyst, nerve damage, and re-tethering in the observation groups were significantly lower than those in the control group ( F=6.395, P<0.05). Conclusion:HBO combined with microsurgical lysis is effective in the treatment of lipoma-type TCS, which can improve the anoxic-ischemic state of spinal cord neurons, accelerate the tibial nerve conduction velocity, significantly improve the comprehensive score of SBNS, and significantly reduce the incidence of postoperative complications. Thus, it is worthy of clinical application.

Objective To explore the key initiatives and effective methods for preparing the constructions of emergency-oriented hospitals under COVID-19 pandemic. Methods The wartime mechanism was strengthened by adhering to unified leadership, trengthening the top-level design and clarifying the division of responsibilities. Objective management was used as a means to take into account the key of personnel allocation and training, prevention and control of hospital infection, transformation of contagious ward, logistic support, equipment and material supply and construction of system and process. Results The preparations and constructions of the emergency-oriented hospitals were completed in 72 hours，which passed the acceptance and inspections from infection control experts，who appraised our work to be “the highest in difficulty, the fastest in project progress and the highest in quality". Totally, upon to the preparations，14 medical teams were set up and the layout process reestablishment of 14 wards was completed, the installation and preparation of nearly 10000 sets/pieces of medical equipment and medical materials were completed as well and more than 80 work systems and process systems for 9 major modules were established. Conclusion The preparations and constructions of emergency-oriented hospitals should be performed upon the thorough implementation of the decisions and arrangements by the municipal Party committee and the municipal government, insisting on the wartime thinking and establishment of high-quality management team and effective goal management focusing on details and actual needs of medical staff.

Objective: To study the related factors of radiation induced lung injury(RILI) after three-dimensional conformal radiotherapy(3D-CRT) in the elderly patients with advance esophageal cancer, thus to provide reference for radiotherapy planning. Methods: The clinical data and physical parameters of 72 elderly patients with advance esophageal cancer were analyzed, including gender, age, performance status scoring, smoking history, tumor location, T staging, underlying disease of the lung, radiation dose, the whole lung accepted 5 Gy exposure volume(V5), V10, V15, V20, V25, V30, V40 and mean lung dose(MLD). Results: Among 72 patients, 15 cases developed RILI.It was revealed by univariate analysis that there were statistically significant differences in advanced age, underlying disease of the lung, V5, V10, V15, V20, V25, MLD with RILI(χ²=5.098, P=0.026; χ²=3.598, P=0.030; t=3.854, P=0.034; t=4.901, P=0.022, t=4.638, P=0.029, t=5.122, P=0.015, t=3.652, P=0.041, t=5.760, P=0.010). Conclusion It should be payed more attention to factors such as advanced age, underlying disease of the lung, V5, V10, V15, V20, V25 and MLD when elderly patients with advance esophageal cancer patients were treated with 3D-CRT in order to prevent and decrease the risks of RILI.

Objective To establish a simple ,low cost and stable method to detect right ventricular pressure in mice .Methods A PE-50 duct length 15 cm(outside diameter :0 .9 mm ,inner diameter :0 .5 mm) was bent on one terminal and the other terminal was inserted into a 7# syringe needle to connect to a pressure transducer .This duct was intubated into right ventricle via right external jugular vein to detect right ventricular pressure in 80 SPF grade male C57BL/6 mice .Successful cases and operation time were re-corded .Besides ,40 SPF grade male C57BL/6 mice were randomized into the control group (n=20) and chronic hyperbaric hypoxia group(n= 20) .Mice in chronic hyperbaric group were raised in a hyperbaric chamber of simulated 5 000 m high altitude for 4 weeks .The control group was raised outside the chamber simultaneously .Right ventricular systolic pressure was detected with the PE duct .Left and right ventricles were detached and weighed ,and Hermann-Willson index was calculated .Results With this PE duct ,right ventricular intubation success rate was 90% (72/80) ,the operation cost approximately 3 to 5 min each mouse from the separation of blood vessels to detect the time needed for the right ventricle waveform .right ventricular systolic pressure[(39 .52 ± 4 .34 )mm Hg] and Hermann-Willson index(0 .356 ± 0 .039)of chronic hyperbaric hypoxia group were significantly higher than that of control group [(21 .24 ± 2 .7)mm Hg and (0 .256 ± 0 .020)] ,which has significant positive correlation (P<0 .01) .Conclusion It is simple ,fast ,stable ,costing low and of high success ratio to detect right ventricular pressure with this method .