OBJECTIVES: To observe the effect of 4-(arylethynyl)-pyrrolo[2,3-d] pyrimidine (10b) on post-traumatic stress disorder (PTSD)-like behaviors and ERK1/2-SGK1 signaling pathway in mice. METHODS: C57BL/6 mouse models exposed to single prolonged stress (SPS) were treated with daily gavage of saline, 10b at low, moderate and high doses, or paroxetine for 14 days. The changes in PTSD-like behaviors of SPS mice with different treatments were observed using behavioral tests. Western blotting and immunofluorescence assay were used to detect the protein expression levels of mGluR5, p-ERK, and SGK1 in the hippocampus of the mice. Pathological changes in the liver and kidney tissues of the mice were examined using HE staining. Molecular docking and molecular dynamics analyses were employed to evaluate the binding stability between the compound 10b and mGluR5. RESULTS: Compared to the normal control mice, the SPS mice exhibited obvious PTSD-like behaviors with increased hippocampal expressions of mGluR5 and p-ERK proteins and decreased SGK1 protein expression. Compound 10b significantly ameliorated behavioral abnormalities in SPS mice, inhibited mGluR5 expression, and reversed the dysregulation of p-ERK and SGK1. No obvious liver or kidney toxicity was observed after 10b treatment. Molecular docking and dynamics studies demonstrated a stable interaction between 10b and mGluR5. CONCLUSIONS The compound 10b ameliorates PTSD-like behaviors induced by SPS in mice possibly by inhibiting mGluR5 expression to modulate the ERK1/2-SGK1 signaling pathway.
Animals ; Stress Disorders, Post-Traumatic/drug therapy* ; Receptor, Metabotropic Glutamate 5/metabolism* ; Mice, Inbred C57BL ; Mice ; Protein Serine-Threonine Kinases/metabolism* ; Pyrimidines/pharmacology* ; Immediate-Early Proteins/metabolism* ; Signal Transduction/drug effects* ; MAP Kinase Signaling System/drug effects* ; Male ; Molecular Docking Simulation ; Hippocampus/metabolism*

Objective To study whether emodin,a natural product,can affect the level of histone acetylation in HpG2 hepatocellular carcinoma cells(HCC),and then accelerate the occurrence of pyroptosis and apoptosis in hepatocellular carcinoma cells,so as to provide a new target for the treatment of liver cancer.Methods CCK-8 method was used to detect the effect of different concentrations of emodin on the viability of HpG2 cells.Bioinformatics was used to analyze histone acetylation-related genes in patients with liver cancer in TCGA database.The correlation between the candidate gene lysine acetyltransferase 2A(KAT2A)and the apoptosis pathway was verified.qPCR method was used to detect the mRNA level of KAT2A in HepG2 cells and L02 cells.The effects of emodin on histone acetyltransferase(HAT)and histone deacetyltransferase(HDAC),interleukin 1β(IL-1β)and interleukin 18(IL-18)in HpG2 cells were detected by ELISA.The effect of emodin on the apoptosis of liver cancer cells was detected by flow cytometry.The expression level of cell apoptosis,pyroptosis-associated protein B lymphocytoma-2(Bcl-2),Bcl-2-related X protein(Bax),NOD-like receptor thermal protein domain-related protein 3(NLRP3),Caspase-1,Gasdermin family member DN terminal(GSDMD-N)and KAT2A were detected by Western blot assay.Results Emodin could reduce the activity of HpG2 cells,and the confidence interval of IC5095%was 58.12-66.52μmol/L.Compared with normal liver tissue,the expression of histone acetylation related gene mRNA was increased in HCC tissue,and the change of KAT2A was the highest[log2(Fold Change)=2.010,P＜0.01].In HCC tissue,the expression of KAT2A mRNA was negatively correlated with apoptosis pathway(rs=-0.230,P＜0.01).Compared with L02 cells,the expression of KAT2A mRNA in HepG2 was higher(P＜0.05).Compared with the control group,expression levels of HAT and HDAC decreased in the 60μmol/L emodin intervention group,expression levels of IL-18 and IL-1β increased,the apoptosis rate increased,expression levels of KAT2A and BAX decreased,and expression levels of Bcl-2,NLRP3,GSDMD-N and Caspase-1 increased(P＜0.05).Conclusion Emodin could inhibit the viability of hepatoma cells,accelerate apoptosis and pyroptosis,and its mechanism may be related to the regulation of KAT2A expression.

Objective:To analyze the genetic mutation characteristics of glucose-6-phosphate dehydrogenase (G6PD) deficiency among infants in Kunming.Methods:A total of 15 533 infants (7 994 males and 7 539 females) born in Kunming from January 1, 2018, to December 31, 2020, with an age range of 2 to 44 days, were selected. G6PD enzyme activity and gene mutation types were detected using fluorescence quantitative analysis, multicolor melting curve analysis (MMCA), and Sanger sequencing. Droplet digital PCR (ddPCR) was used for quantitative analysis of a newly identified variant family to determine the mutant allele proportion in family members. Meanwhile,the protein structure model and pathogenicity prediction of the novel variant were analyzed.Data analysis was conducted using SPSS 26.0. Specifically, chi-square tests were used for the detection rates of G6PD enzyme activity and gene mutations between different genders. One-way analysis of variance (ANOVA) was used for the comparison of enzyme activity among different mutation types.Results:Among 15 533 infants, 143 cases (129 males and 14 females) were tested positive for G6PD activity, with a detection rate of 0.92% (143/15 533). The difference in detection rates between males and females was statistically significant (χ 2=96.76, P<0.001). Out of 89 enzyme activity-positive cases (83 males and 6 females) underwent genetic testing, 77 (72 males and 5 females) were detected by MMCAand other 12 negative samples were underwent further Sanger sequencing, revealing mutations in 6 samples, all of which were males. Among the 83 individuals with gene mutations, 78 had heterozygous mutations, 1 had a homozygous mutation, and 4 had compound heterozygous mutations. A total of 12 mutation types were detected, with G6PD c.487G>A, c.1024C>T, c.1388G>A, and c.1376G>T being the most common, accounting for 74.70% (62/83) of all mutation types. The average G6PD enzyme activity of c.1376G>T was the lowest, and the differences were statistically significant compared to the average enzyme activity of the other three mutations ( P<0.05). One male infant with a newly identified G6PD c.242G>C mutation was detected, predicted to be pathogenic. ddPCR confirmed that the mother of the affected child was a c.242G>C mutant chimera, with a chimera proportion of 6.66%. Conclusions:In the Kunming region, the predominant G6PD deficiency gene mutation is c.487G>A, with the detection of a novel G6PD c.242G>C mutation. The application of ddPCR technology can assist in detecting the proportion of mutation chimeras.

Objective:To explore the predictive value of systemic immune-inflammation index (SII) and small and dense low-density lipoprotein-cholesterol (sdLDL-C) on contrast-induced acute kidney injury (CI-AKI) in patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary intervention (PCI).Methods:This retrospective analysis included 674 STEMI patients who underwent emergency PCI in Affiliated Hospital of Xuzhou Medical University from November 2019 to October 2021, all patients were divided into a training cohort ( n=450) and validation cohort ( n=224) at a ratio of 2∶1 according to the chronological sequence. The patients in the training cohort were further divided into CI-AKI group ( n=92) and non-CI-AKI group ( n=358). Information at admission and emergency blood biochemical indexes were collected, and the SII was calculated. Multifactorial logistic regression analysis was used to explore the independent factors influencing the occurrence of CI-AKI in STEMI patients undergoing emergency PCI in the training cohort and a predictive model was established. Receiver operating characteristic (ROC) curve and Hosmer-Lemeshow test were used to evaluate the model discrimination and calibration. Results:The prevalence of CI-AKI was 20.4% (92/450). Age, proportion of women, sdLDL-C, urea, baseline creatinine, uric acid, neutrophil count, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and SII were significantly higher in the CI-AKI group than in the non-CI-AKI group (all P<0.05), and left ventricular ejection fraction (LVEF), high-density lipoprotein cholesterol, estimated glomerular filtration rate (eGFR) and lymphocyte count were significantly lower in the CI-AKI group than in the non-CI-AKI group (all P<0.05). The results of multifactorial logistic regression analysis showed that age ( OR=1.046, P=0.001), LVEF ( OR=0.916, P<0.001), sdLDL-C ( OR=4.754, P<0.001), uric acid ( OR=1.012, P=0.007), eGFR ( OR=0.994, P=0.002), and lnSII ( OR=2.471, P<0.001) were independent determinants of CI-AKI after emergency PCI in STEMI patients. ROC curve analysis showed that area under the curve (AUC) for the diagnosis of CI-AKI was 0.688 with a sensitivity of 73.9% and specificity of 61.5% for the SII cut-off point of 1 179.07×10 9/L. The AUC for the diagnosis of CI-AKI was 0.709 with a sensitivity of 65.2% and specificity of 77.4% for the sdLDL-C cut-off point of 1.147 mmol/L. The AUC for the diagnosis of CI-AKI was 0.847 with a sensitivity of 88.0% and a specificity of 70.6% for the combination of SII and sdLDL-C with age, LVEF, uric acid and eGFR. The Hosmer-Lemeshow test (χ2=6.913, P=0.546) proved the goodness of fit of the model. Conclusions:SII and sdLDL-C have significant clinical value in the prediction of CI-AKI. SII and sdLDL-C combined with age, LVEF, uric acid and eGFR could further improve the predictive efficacy of CI-AKI.

Rho-associated kinases (ROCKs) are serine-threonine protein kinases that act downstream of small Rho GTPases to regulate the dynamics of the actin cytoskeleton. Two ROCK isoforms (ROCK1 and ROCK2) are expressed in the mammalian central nervous system. Although ROCK activity has been implicated in synapse formation, whether the distinct ROCK isoforms have different roles in synapse formation and function in vivo is not clear. Here, we used a genetic approach to address this long-standing question. Both Rock1 and Rock2 mice had impaired glutamatergic transmission, reduced spine density, and fewer excitatory synapses in hippocampal CA1 pyramidal neurons. In addition, both Rock1 and Rock2 mice showed deficits in long-term potentiation at hippocampal CA1 synapses and were impaired in spatial learning and memory based on the water maze and contextual fear conditioning tests. However, the spine morphology of CA1 pyramidal neurons was altered only in Rock2 but not Rock1 mice. In this study we compared the roles of ROCK1 and ROCK2 in synapse formation and function in vivo for the first time. Our results provide a better understanding of the functions of distinct ROCK isoforms in synapse formation and function.

'Three-acupoint and Five-needle Method' is a summary of Prof. Shao Jing-ming's clinical experience in the treatment of asthma for years. It is used to alleviate asthma during seizure and to improve the pulmonary functions, strengthen the body constitution and prevent recurrence in the remission stage. This article introduces the theoretic source and basic principle, and concrete operating method, i.e. puncturing bilateral Feishu (BL 13), Dazhui (GV 14) and bilateral Fengmen (BL 12), and adding acupoints based upon pattern identification, in combination of moxibustion and cupping therapy for promoting the clinical application of this method.

Objective To study the therapeutic effect of IGF?1R inhibitor TAE226 on malignant pleural effusion ( MPE) in nude mice. Methods Human lung carcinoma A549 cells were injected into the pleural cavity of nude mice to establish MPE model. The mice were randomly divided into model group and treatment group, and were orally administered with distilled water and TAE226 ( 20 mg/kg ) in the same volume, respectively. The volume of pleural effusion and tumor weight of the two groups were observed. HE staining was used to reveal the histological changes and enzyme?linked immunosorbent assay ( ELISA) was used to detect the IGF?1R protein expression. IGF?1R mRNA level in the tumor tissue was determined by RT?PCR. Microvessel density (MVD) and cell proliferation index (PI) were assessed by immunohistochemical analysis. The protein expression levels of IGF?1R, p?IGF?1R, PI3K and p?PI3K in the tumor tissue were determined by Western blotting. Results The volumes of pleural effusion were ( 241. 4 ± 89. 7 ) μl and (121.7±78.8) μl in the model and treatment groups, respectively (P<0.05). The tumor weight of treatment group was (316.7±186.3) mg, significantly lower than that of the model group (671.4±281.4) mg (P<0.05) . RT?PCR analysis showed that IGF?1R mRNA level was 0. 914 ± 0. 029 in the treatment group, significantly lower than that of the model group (1.152±0.037, P<0.01). The ELISA data revealed that IGF?1R protein expression level of the model group was significantly higher than that of the treatment group [(41.0±4.7) μg/L vs. (24.0±3.1) μg/L, P<0.01]. Immunohistochemical analysis showed that there were significant differences between MVD and PI in the model and treatment groups [ MVD, 34. 75 ± 3. 49 vs. 22.25±3.63;PI, (75.25±7.15)% vs. (45.75±5.12)%;P<0.01 for both). Western blot data showed that IGF?1R and PI3K protein expression levels were not significantly different between the model and treatment groups (1.03±0.33 vs. 0.98±0.37 and 1.05±0.28 vs. 0.98±0.19), respectively (P>0.05), but p?IGF?1R and p?PI3K protein expression levels had significant differences between the two groups (1.08±0.10 vs. 0.51± 0.08 and 1.12±0.09 vs. 0.86±0.09), respectively (P<0.01 for both). Conclusions The IGF?1R inhibitor can effectively inhibit the formation of malignant pleural effusion. Its mechanism may be related to the suppression of tumor cell proliferation, invasion and angiogenesis through inhibition of PI3K signaling. TAE226 treatment may be a potential therapeutic regimen of treating malignant pleural effusion.

Objective To study the therapeutic effect of IGF?1R inhibitor TAE226 on malignant pleural effusion ( MPE) in nude mice. Methods Human lung carcinoma A549 cells were injected into the pleural cavity of nude mice to establish MPE model. The mice were randomly divided into model group and treatment group, and were orally administered with distilled water and TAE226 ( 20 mg/kg ) in the same volume, respectively. The volume of pleural effusion and tumor weight of the two groups were observed. HE staining was used to reveal the histological changes and enzyme?linked immunosorbent assay ( ELISA) was used to detect the IGF?1R protein expression. IGF?1R mRNA level in the tumor tissue was determined by RT?PCR. Microvessel density (MVD) and cell proliferation index (PI) were assessed by immunohistochemical analysis. The protein expression levels of IGF?1R, p?IGF?1R, PI3K and p?PI3K in the tumor tissue were determined by Western blotting. Results The volumes of pleural effusion were ( 241. 4 ± 89. 7 ) μl and (121.7±78.8) μl in the model and treatment groups, respectively (P<0.05). The tumor weight of treatment group was (316.7±186.3) mg, significantly lower than that of the model group (671.4±281.4) mg (P<0.05) . RT?PCR analysis showed that IGF?1R mRNA level was 0. 914 ± 0. 029 in the treatment group, significantly lower than that of the model group (1.152±0.037, P<0.01). The ELISA data revealed that IGF?1R protein expression level of the model group was significantly higher than that of the treatment group [(41.0±4.7) μg/L vs. (24.0±3.1) μg/L, P<0.01]. Immunohistochemical analysis showed that there were significant differences between MVD and PI in the model and treatment groups [ MVD, 34. 75 ± 3. 49 vs. 22.25±3.63;PI, (75.25±7.15)% vs. (45.75±5.12)%;P<0.01 for both). Western blot data showed that IGF?1R and PI3K protein expression levels were not significantly different between the model and treatment groups (1.03±0.33 vs. 0.98±0.37 and 1.05±0.28 vs. 0.98±0.19), respectively (P>0.05), but p?IGF?1R and p?PI3K protein expression levels had significant differences between the two groups (1.08±0.10 vs. 0.51± 0.08 and 1.12±0.09 vs. 0.86±0.09), respectively (P<0.01 for both). Conclusions The IGF?1R inhibitor can effectively inhibit the formation of malignant pleural effusion. Its mechanism may be related to the suppression of tumor cell proliferation, invasion and angiogenesis through inhibition of PI3K signaling. TAE226 treatment may be a potential therapeutic regimen of treating malignant pleural effusion.

Adult ; Branchial Region ; Fistula ; surgery ; Humans ; Male ; Neck ; surgery ; Pharynx ; surgery

OBJECTIVETo study the protective effect and mechanism of salvianolic acid B (Sal B) on glutamate-induced excito-toxicity.

METHODGlutamate-induced PC12 cell injury model was established to detect the cell survival rate by MTT, the leakage rate of lactic dehydrogenases using LDH, and the cell apoptosis by using AO/EB double staining for fluorescence microscope and PI single staining flow cytometry which was also used to detect the content of intracellular reactive oxygen species. The expression of Caspase-3 protein was also detected by the Western blotting method.

RESULTSal B is proved to inhibit glutamate-induced PC12 cells from injury and prevent them from releasing LDH within the range from 50 micromol x L(-1) to 200 micromol x L(-1). Meanwhile, Sal B has an effect on significantly reducing the expression of inhibit glutamate-induced active Caspase-3 protein, inhibiting accumulated glutamate-induced ROS and decreasing PC12 cell apoptosis rate within the range from 50 micromol x L(-1) to 200 micromol x L(-1).

CONCLUSIONThe study proves that Sal B prevented against glutamate-induced cell injury via inhibiting ROS formation and Caspase-3 pathway-dependent apoptosis in PC12 cells.

Action Potentials ; drug effects ; Animals ; Apoptosis ; drug effects ; Benzofurans ; pharmacology ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Excitatory Amino Acid Antagonists ; pharmacology ; Glutamic Acid ; adverse effects ; Lactate Dehydrogenases ; metabolism ; PC12 Cells ; Pheochromocytoma ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism