BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

OBJECTIVES: To explore the therapeutic mechanism of Chaihu Shugan (CHSG) Decoction for improving cognitive impairment in rats with epilepsy induced by lithium chloride and pilocarpine. METHODS: Male SD rat models of cognitive impairment model after epilepsy induced by intraperitoneal injection with lithium chloride and pilocarpine were randomly divided into 5 groups (n=12) for treatment with daily gavage of saline, donepezil (90 mg/kg), or CHSG Decoction at 2.5, 5.0, 10, 20 and 40 g/kg for 4 consecutive weeks, with 10 rats with intraperitoneal injection with saline as the blank control group. Morris water maze test was used to evaluate cognitive and behavioral changes of the rats after treatment. The mRNA and protein expressions of ASIC1, NR1, NR2A and NR2B in the hippocampus of rats were detected using RT-qPCR and Western blotting. RESULTS: Compared with those with saline treatment, the rat models treated with CHSG Decoction at 5 and 10 g/kg showed significantly shortened escape latency and prolonged stay in the target quadrant with increased number of platform crossings in Morris water maze test. CHSG Decoction treatment at the two doses significantly increased ASIC1, NR1, NR2A and NR2B protein expressions in the hippocampus of the rat models, and their mRNA expression levels were all increased significantly after the treatment at the doses above 2.5 g/kg. CONCLUSIONS CHSG Decoction can improve cognitive impairment in rats after epilepsy possibly by regulating the expression and channel activity of NMDAR protein and its subunit protein via upregulating ASIC1 to modulate neuronal excitability and synaptic plasticity in the hippocampus.
Animals ; Hippocampus/drug effects* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Acid Sensing Ion Channels/metabolism* ; Rats, Sprague-Dawley ; Male ; Rats ; Epilepsy/complications* ; Cognitive Dysfunction/drug therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Up-Regulation ; Maze Learning

Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

Objective To explore the therapeutic mechanism of Chaihu Shugan(CHSG)Decoction for improving cognitive impairment in rats with epilepsy induced by lithium chloride and pilocarpine.Methods Male SD rat models of cognitive impairment model after epilepsy induced by intraperitoneal injection with lithium chloride and pilocarpine were randomly divided into 5 groups(n=12)for treatment with daily gavage of saline,donepezil(90 mg/kg),or CHSG Decoction at 2.5,5.0,10,20 and 40 g/kg for 4 consecutive weeks,with 10 rats with intraperitoneal injection with saline as the blank control group.Morris water maze test was used to evaluate cognitive and behavioral changes of the rats after treatment.The mRNA and protein expressions of ASIC1,NR1,NR2A and NR2B in the hippocampus of rats were detected using RT-qPCR and Western blotting.Results Compared with those with saline treatment,the rat models treated with CHSG Decoction at 5 and 10 g/kg showed significantly shortened escape latency and prolonged stay in the target quadrant with increased number of platform crossings in Morris water maze test.CHSG Decoction treatment at the two doses significantly increased ASIC1,NR1,NR2A and NR2B protein expressions in the hippocampus of the rat models,and their mRNA expression levels were all increased significantly after the treatment at the doses above 2.5 g/kg.Conclusion CHSG Decoction can improve cognitive impairment in rats after epilepsy possibly by regulating the expression and channel activity of NMDAR protein and its subunit protein via upregulating ASIC1 to modulate neuronal excitability and synaptic plasticity in the hippocampus.

Objective To establish an optimal limiting dose for dose limiting rings placed in the anterior and posterior regions of the neck for reducing head and neck lymphedema under intensity-modulated radiation therapy(IMRT)for early nasopharyngeal carcinoma(NPC)from a dosimetric perspective.Method Fifteen newly diagnosed early-stage nasopharyngeal carcinoma patients who underwent CT localization for radiotherapy at the Cancer Hospital of Guangzhou Medical University from January to September 2022 were included in the study.Each case was designed with five sets of radiotherapy plans.Plan A consisted of conventional unlimited-field plans,while Plans B-E consisted of limited-field plans with dose constraints set at 20,18,16,and 14 Gy,respectively,with the remaining parameters consistent with Plan A.The impact on target coverage and organ-at-risk constraints was evaluated through variance analysis and pairwise multiple comparisons using a randomized block design to determine the optimal dose limits.Results The gradient of 16Gy was determined as the optimal dose limiting cutoff point for achieving the balance between target coverage and organ limiting dose.Compared with the conventional plan,The plans with the placement of a cervical anterior and posterior dose limiting ring(16Gy)did not change the target dose coverage(P>0.05),but only yielded a slight change in the homogeneity index(P<0.05).It did not cause any changes of the dosage in the inner ear,mandible,and brainstem(all P>0.05),but lead to statisti-cally significant reductions in the oral cavity,throat,and thyroid(all P<0.05).It caused a slight increase of the dose in the parotid gland and spinal cord(both P<0.05),but the increased dose was anyhow within the tolerance range.Conclusion The dosimetric investigation determines an optimal dose limit cutoff point for the cervical ante-rior and posterior dose limiting rings.It is expected to provide a design method for IMRT plans to reduce head and neck lymphedema after radiotherapy for early NPC.

Objective:To investigate the related factors of pleasure loss in patients with human immunodefi-ciency virus(HIV)/acquired immune deficiency syndrome(AIDS).Methods:Totally 237 patients with HIV/AIDS from a certain infectious disease hospital were selected and surveyed with a self-designed general information ques-tionnaire,the Temporal Pleasure Experience Scale(TEPS),Self Acceptance Scale(SAQ),Discrimination Percep-tion Scale(SIS),and Perceived Social Support Scale(PSSS).Results:The patient's TEPS score was(73.4±16.1).Stepwise linear regression analysis showed that the PSSS total scores,education level,and personal monthly income were positively correlated with the TEPS total scores(β=0.41,5.17,4.63),and age was negatively corre-lated with the TEPS total scores(β=-0.30).Conclusion:It suggests that more attention should be paid to the lack of pleasure in patients with HIV/AIDS,and the lack of pleasure is related to personal monthly income,educa-tion level,age and perceived social support.

Objective:To explore the optimal insertion point of reverse sacral alar-iliac screws (RSAIS) in sacroiliac joint fixation for Tile type C pelvic fracture and the effectiveness of its preliminary clinical application.Methods:CT data from 90 patients with no abnormal pelvic structures in the Radiology Department database of Liyang People's Hospital from January 2022 to January 2023 were selected. There were 45 males and 45 females, aged 21-69 years [(45.5±6.4)years]. With CT digital reconstruction technology, the RSAIS fixation was simulated and the anterior insertable area was measured. Five insertion points were at the ilium-acetabular recess in the area superior posterior to the acetabulum (A1), at 1 cm (A2) and 2 cm (A3) posterior transversal to A1, at 1 cm (A4) and 2 cm (A5) inferior longitudinal to A1. With the entry point of S 1 pedicle screw as the exit point, the transverse and sagittal CT scans were conducted on 5 screw trajectories of anterior posterior lines (corresponding to five groups). The anatomical parameters of the screw trajectories, including the length, width and height of the screw trajectories were measured by two observers independently and the intraclass correlation coefficient (ICC) between the observers was analyzed. Tile type C pelvic fracture models were established from 20 3D-printed L 5-pelvis-femur specimens and divided into 5 groups, with 4 specimens in each group. The anterior rings were fixed with cross screws while the posterior rings with one RSAIS in each group using one of the aforementioned 5 screw trajectories. The overall displacement and local displacement of the sacroiliac space under 300 N vertical load in all the groups were measured with a biomechanical machine. The screw trajectory was designed according to the optimal insertion point shown in the above anatomical measurements and mechanical experiments. An RSAIS with a diameter of 7.3 mm was inserted percutaneously to treat a patient with Tile type C1 pelvic fracture. The fluoroscopy time of screw insertion and screw position were recorded as well as bone reunion, numerical rating scale for pain, Oswestry dysfunction index (ODI) and Majeed functional score of pelvic fracture at the last follow-up. Results:The anterior insertable area was (1236.64±12.04)mm 2. The trajectory lengths for the three transversal insertion points gradually decreased from front to back, with A1 being the longest (104.9±10.4)mm, followed by A2 (98.5±9.8)mm and A3 the shortest (92.7±9.7)mm ( P<0.01), while there were no significant differences in the widths or heights ( P>0.05). There were no significant differences in the length, width or height of the screw trajectories of the three longitudinal insertion points (groups A1, A4 and A5) ( P>0.05). Compared with A4 and A5, A1 was farther from the margin of the acetabulum. The range of ICC of the anatomical parameters of the 3 screw trajectories measured by the two observers was 0.88-0.98, 95% CI 0.90, 0.96, and 95% CI 0.95, 0.99, indicating high consistency. After the screws were placed and fixed at the 3 transversal insertion points, the overall displacement and local displacement of the sacroiliac space of groups A1, A2 and A3 gradually increased from front to back. They were (2.93±0.09)mm and (1.49±0.14)mm in group A1, (3.14±0.12)mm and (1.63±0.54)mm in group A2 and (3.23±0.12)mm and (1.67±0.67)mm in group A3 respectively. There was a significant difference in the overall displacement among three groups ( P<0.01), while the local displacement of the sacroiliac space in group A1 was decreased compared with that of group A2 and A3 ( P<0.01). After the screws were placed and fixed at the 3 longitudinal insertion points, there were no significant differences in the overall displacement and local displacement of the sacroiliac space of group A1, A4 and A5 ( P>0.05). The fluoroscopy time of RSAIS inserted from A1 for the treatment of patients with Tile C1 pelvic fracture was 66 seconds, with fine screw position. At the last follow-up, the fracture was healed, with numerical rating scale for pain decreased from 6 points preoperatively to 1, ODI improved from 41 preoperatively to 18, and the Majeed functional score of 81 points. Conclusions:For Tile type C pelvic fracture, the screw trajectory from the iliac-acetabular recess located in the superior anterior part of the insertable area is the longest, with the best stability and relatively good safety of the screw, making it the optimal insertion point for RSAIS. The effect of preliminary clinical application of RSAIS from the optimal insertion point is satisfactory.

High-altitude regions,characterized by their elevated altitude,are subject to a complex set of environmental conditions including intense ultraviolet radiation,low oxygen levels,low temperatures,and low humidity.These distinctive environmental features lead to unique dietary patterns,lifestyles,and physiological adaptations.Notably,individuals who have just moved into high-altitude areas and those who live there on a long-term basis undergo specific adaptive adjustments in glucose metabolism.Typically,newcomers experience transient elevations in blood glucose levels,which gradually decline after prolonged residence at high altitudes to levels even lower than those found at low altitudes.In general,current findings of observational studies generally suggest a decreased risk of diabetes mellitus among populations inhabiting high-altitude regions.However,the glucose metabolism varies among populations from different high-altitude regions across the world,which indicates that the reshaping of glucose metabolism induced by high altitudes is a complicated phenomenon.This article provides an overview of the impact of various components of high-altitude environment,characteristic lifestyle factors,and socioeconomic development levels on glucose metabolism and the related diseases and the potential mechanisms involved.The aim is to offer valuable insights for researchers investigating glucose metabolism in high-altitude settings.

Objective:To explore the predictive value of peripheral blood cytokine models on organ functional impairment after chimeric antigen receptor T(CAR-T) cell therapy in children with B-lineage lymphocytic leukemia.Methods:The clinical data of 44 children with acute B-lineage lymphoblastic leukemia who received CAR-T cell therapy at Children′s Hospital of Soochow University from September 2018 to October 2020 were retrospectively analyzed.Peripheral blood cytokines, including interleukin(IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor-α, interferon(IFN)-γ and IL-17A, were measured daily for 14 days after receiving CAR-T cell therapy.The trend of peripheral blood cytokine levels was analyzed at the endpoint of organ function recovery or death within 14 days after CAR-T cell treatment.Receiver operating characteristic curve was used to establish a mathematical prediction model to predict the occurrence of organ damage in the children.Results:Of the 44 children, 31 cases were boys and 13 cases were girls, with a median age of 7.96 (5.19, 11.48)years.Cytokine release syndrome(CRS) response occurred in 95.5% (42/44) children, with 88.1% (37/42) had a grade 1-3 CRS response, and 16.7% (7/42) had a severe grade 4-5 CRS response.Using IL-6>3 892.95 pg/mL as cut-off value, the area under the curve(AUC) for predicting acute respiratory failure was 0.818, with a sensitivity of 0.8 and a specificity of 0.735, while combining IFN-γ>414.4 pg/mL, IL-6>3 892.95 pg/mL and IL-2>27.05 pg/mL were the three cut-off values, with an AUC of 0.741, sensitivity of 0.6 and specificity of 0.912 for predicting acute respiratory failure. Using IFN-γ>1 699.5 pg/mL as cut-off value, the AUC for predicting shock was 0.908, with a sensitivity of 0.722 and a specificity of 1.With IL-6>4 607.3 pg/mL as cut-off value, the AUC for predicting liver injury was 0.964, with a sensitivity of 1 and a specificity of 0.906, while combining both IL-6>4 607.3 pg/mL and IFN-γ>1 446.2 pg/mL as cut-off values, the AUC for predicting liver injury was 0.977, with a sensitivity of 1 and a specificity of 0.906.Combining both IL-6>6 972.2 pg/mL and IFN-γ>3 981.5 pg/mL predicted a positive predictive value of 62.5% and a negative predictive value of 94.4% for grade 4-5 CRS response, with an AUC of 0.846, a predictive sensitivity of 0.714 and a specificity of 0.838, and all children had a combination of two or more organ function injuries.Conclusion:The combination of IL-6 and IFN-γ can effectively predict the incidence of liver injury and cytokine release syndrome.The combination of peripheral blood cytokines IFN-γ, IL-6 and IL-2 can be used to predict the incidence of acute respiratory failure after the treatment of CAR-T cells in children with acute B-lineage lymphoblastic leukaemia.IFN-γ single index can be used to predict the incidence of shock.The combination of IL-6 and IFN-γ can be used to predict the incidence of liver injury and the severity of CRS.

Objective:To investigate the value of monitoring on serum silent information regulator-related enzyme 3 (SIRT3), glucagon-like peptide-1 (GLP-1) and angiopoietin-like protein 4 (ANGPTL4) in patients with acute ischemic stroke (AIS).Methods:Eighty patients with AIS who treatment in Qiongzhong Li and Miao Autonomous County People′s Hospital from May 2019 to April 2022 were selected retrospectively as the observation group, and 60 healthy volunteers who underwent physical examination during the same period were selected as the normal control group. The levels of serum SIRT3, GLP-1, and ANGPTL4 between the two groups were compared. The neurological deficit degree of AIS patients was evaluated by National Institutes of Health Stroke Scale(NIHSS) and the correlation of SIRT3, GLP-1 and ANGPTL4 with neurological deficit degree were analyzed. The levels of serum SIRT3, GLP-1 and ANGPTL4 before and after treatment and their difference value were compared between different clinical outcome of AIS patients, the risk factors for poor clinical outcome of AIS patients were analyzed by Logistic regression analysis, the value of prediction was analyzed by receiver operating characteristic (ROC) curve.Results:The level of serum GLP-1 in the observation group was lower than that in the normal control group: (50.37 ± 5.69) nmol/L vs. (34.89 ± 4.26) nmol/L; and the levels of serum SIRT3 and ANGPTL4 in the observation group were higher than those in the normal control group: (50.37 ± 5.69) ng/L vs. (34.89 ± 4.26) ng/L, (15.07 ± 3.12) μg/L vs. (11.15 ± 2.63) μg/L, there were statistical differences ( P<0.05). The results of correlation analysis showed that the levels of serum SIRT3 and ANGPTL4 were positively correlated with the degree of neurological impairment in AIS patients( r = 0.631, 0.776, P<0.05), and the level of serum GLP-1 was negatively correlated with the degree of neurological impairment in AIS patients ( r = - 0.693, P<0.05). After treatment, 66 patients obtained good clinical outcome, the good outcome rate was 82.50%(66/80). The levels of serum SIRT3 and ANGPTL4 in the poor clinical outcome patients were higher than those in the good clinical outcome patients: (41.33 ± 4.74) ng/L vs. (37.82 ± 4.05) ng/L, (12.98 ± 2.17) μg/L vs. (11.69 ± 2.06) μg/L; the level of serum GLP-1 in the poor clinical outcome patients was lower than that in the good clinical outcome patients: (592.33 ± 98.44) nmol/L vs. (709.41 ± 125.31) nmol/L; the difference value of SIRT3, GLP-1 and ANGPTL4 before and after treatment in the poor clinical outcome patients were lower than those in the good clinical outcome patients: (10.22 ± 2.05) ng/L vs. (12.31 ± 2.94) ng/L, (268.21 ± 70.12) nmol/L vs. (379.92 ± 85.33) nmol/L, (2.18 ± 0.65) μg/L vs. (3.36 ± 0.94) μg/L, there were statistical differences ( P<0.05). The results of Logistic regression analysis showed that differences value of SIRT3, GLP-1 and ANGPTL4 before and after treatment were all independent influencing factors of poor clinical outcome in patients with AIS ( P<0.05). The results of ROC curve analysis showed that the area under the curve (AUC) of differences value of SIRT3, GLP-1 and ANGPTL4 before and after treatment in predicting poor clinical outcome were 0.701, 0.758 and 0.844, respectively, and had certain predictive value, the AUC of joint evaluation was the largest (0.912). Conclusions:The levels of serum SIRT3 and ANGPTL4 in patients with AIS are increased, and the level of serum GLP-1 is decreased, and they are related to the degree of neurological deficit. Clinical monitoring of their level changes is helpful for clinical evaluation of the clinical outcome of patients with AIS.