Objective To explore the medication patterns as well as the potential mechanism of action of traditional Chinese medicine in the treatment of presbycusis based on data mining and network pharmacology.Methods Search for the literatures on the treatment of presbycusis with traditional Chinese medicine in CNKI,Wanfang Data Knowledge Service Platform and VIP,conduct visual analyses such as association rule analysis and cluster analysis on them,and summarize the medication patterns and core drug combinations.Predict the action targets of drugs and diseases through network pharmacology,and obtain core targets and pathways.Results A total of 22 literatures were included,and 22 traditional Chinese medicine prescriptions were obtained,involving 72 kinds of traditional Chinese medicines,with a total frequency of administration of 238 times.The four natures of the medicine were mainly warm,the five flavors were mainly sweet,bitter and pungent,and channel tropism were mainly liver meridian and kidney meridian.The core drug combination was Shanzhuyu and Shudihuang.Network pharmacological analysis showed that this combination may improve presbycusis by influencing core targets such as protein kinase B(Akt)1 and interleukin-6,thereby intervening in related pathways such as advanced glycation end product/advanced glycation end product receptor signaling pathway and the phosphoinositide 3-kinase/Akt signaling pathway.Conclusion The treatment of presbycusis with traditional Chinese medicine mainly focuses on invigorating kidney for consolidating semen,while also circulate and nourish blood.Network pharmacology predicts the key mechanisms of core high-frequency drug treatment for presbycusis,providing data support and theoretical basis for the clinical treatment of traditional Chinese medicine.

Objective To study the immune regulatory effect of total flavonoids from houttuynia cordata on herpes simplex keratitis(HSK)mice and preliminarily explore its mechanism.Methods A surgical blade to mark the"#"character on the corneal epithelial layer of Balb/c mice and inoculate with I type herpes simplex virus(HSV-1)was used to construct an HSK model.The successfully identified HSK mice were randomly divided into model group,low(75 mg/kg)and high-dose(300 mg/kg)total flavonoids from houttuynia cordata groups,and high-dose total flavonoids from houttuynia cordata+toll like receptor(TLR)2 agonist Pam3CSK4(0.3 mg/kg)group.A control group was also established with 12 mice in each group.During the modeling and administration period,the corneal lesions in mice were observed daily using a slit-lamp microscope and scored.After administration,HE staining was used to observe the pathological changes of the mouse cornea.ELISA was used to detect the levels of interferon-γ(INF-γ),interleukin-2(IL-2),IL-4,and IL-6 in mouse serum.Western blot was used to detect the protein expression levels of TLR2,myeloid differentiation factor 88(MyD88),nuclear factor-kappaB p65(NF-κB p65)and p-NF-κB p65 in mouse corneal tissue.Results Compared with the control group,the model group mice showed corneal edema,ulcers,and neovascularization,with severe corneal damage and a large number of inflammatory cell infiltration.The serum levels of INF-γ and IL-2 significantly decreased,while the levels of IL-4 and IL-6 significantly increased(P<0.01).The protein expression levels of TLR2 and MyD88 and protein ratios of p-NF-κB p65/NF-κB p65 in corneal tissue significantly increased(P<0.01).Compared with the model group,the low and high dose of total flavonoids from houttuynia cordata groups showed improved corneal injury,the corneal lesion score significantly dereased(P<0.01).The serum levels of INF-γ and IL-2 significantly increased,while the levels of IL-4 and IL-6 significantly decreased(P<0.01).The protein expression levels of TLR2 and MyD88 and protein ratios of p-NF-κB p65/NF-κB p65 in corneal tissue significantly decreased(P<0.01).However,the TLR2 agonist Pam3CSK4 partially reversed the improvement effects of total flavonoids from houttuynia cordata on keratitis and immune imbalance in HSK mice.Conclusion The total flavonoids of houttuynia cordata can effectively improve corneal damage,regulate immune response,and suppress inflammation levels in HSK mice.Its mechanism of action may be related to the regulation of TLR2/NF-κB signaling pathway.

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

Objective To study the immune regulatory effect of total flavonoids from houttuynia cordata on herpes simplex keratitis(HSK)mice and preliminarily explore its mechanism.Methods A surgical blade to mark the"#"character on the corneal epithelial layer of Balb/c mice and inoculate with I type herpes simplex virus(HSV-1)was used to construct an HSK model.The successfully identified HSK mice were randomly divided into model group,low(75 mg/kg)and high-dose(300 mg/kg)total flavonoids from houttuynia cordata groups,and high-dose total flavonoids from houttuynia cordata+toll like receptor(TLR)2 agonist Pam3CSK4(0.3 mg/kg)group.A control group was also established with 12 mice in each group.During the modeling and administration period,the corneal lesions in mice were observed daily using a slit-lamp microscope and scored.After administration,HE staining was used to observe the pathological changes of the mouse cornea.ELISA was used to detect the levels of interferon-γ(INF-γ),interleukin-2(IL-2),IL-4,and IL-6 in mouse serum.Western blot was used to detect the protein expression levels of TLR2,myeloid differentiation factor 88(MyD88),nuclear factor-kappaB p65(NF-κB p65)and p-NF-κB p65 in mouse corneal tissue.Results Compared with the control group,the model group mice showed corneal edema,ulcers,and neovascularization,with severe corneal damage and a large number of inflammatory cell infiltration.The serum levels of INF-γ and IL-2 significantly decreased,while the levels of IL-4 and IL-6 significantly increased(P<0.01).The protein expression levels of TLR2 and MyD88 and protein ratios of p-NF-κB p65/NF-κB p65 in corneal tissue significantly increased(P<0.01).Compared with the model group,the low and high dose of total flavonoids from houttuynia cordata groups showed improved corneal injury,the corneal lesion score significantly dereased(P<0.01).The serum levels of INF-γ and IL-2 significantly increased,while the levels of IL-4 and IL-6 significantly decreased(P<0.01).The protein expression levels of TLR2 and MyD88 and protein ratios of p-NF-κB p65/NF-κB p65 in corneal tissue significantly decreased(P<0.01).However,the TLR2 agonist Pam3CSK4 partially reversed the improvement effects of total flavonoids from houttuynia cordata on keratitis and immune imbalance in HSK mice.Conclusion The total flavonoids of houttuynia cordata can effectively improve corneal damage,regulate immune response,and suppress inflammation levels in HSK mice.Its mechanism of action may be related to the regulation of TLR2/NF-κB signaling pathway.

Objective To explore the medication patterns as well as the potential mechanism of action of traditional Chinese medicine in the treatment of presbycusis based on data mining and network pharmacology.Methods Search for the literatures on the treatment of presbycusis with traditional Chinese medicine in CNKI,Wanfang Data Knowledge Service Platform and VIP,conduct visual analyses such as association rule analysis and cluster analysis on them,and summarize the medication patterns and core drug combinations.Predict the action targets of drugs and diseases through network pharmacology,and obtain core targets and pathways.Results A total of 22 literatures were included,and 22 traditional Chinese medicine prescriptions were obtained,involving 72 kinds of traditional Chinese medicines,with a total frequency of administration of 238 times.The four natures of the medicine were mainly warm,the five flavors were mainly sweet,bitter and pungent,and channel tropism were mainly liver meridian and kidney meridian.The core drug combination was Shanzhuyu and Shudihuang.Network pharmacological analysis showed that this combination may improve presbycusis by influencing core targets such as protein kinase B(Akt)1 and interleukin-6,thereby intervening in related pathways such as advanced glycation end product/advanced glycation end product receptor signaling pathway and the phosphoinositide 3-kinase/Akt signaling pathway.Conclusion The treatment of presbycusis with traditional Chinese medicine mainly focuses on invigorating kidney for consolidating semen,while also circulate and nourish blood.Network pharmacology predicts the key mechanisms of core high-frequency drug treatment for presbycusis,providing data support and theoretical basis for the clinical treatment of traditional Chinese medicine.

Objective:To explore the robust relationship between insomnia and type 2 diabetes mellitus by two-sample Mendelian randomization analysis to overcome confounding factors and reverse causality in observational studies.Methods:We identified strong,independent single nucleotide polymorphisms(SNPs)of insomnia from the most up to date genome wide association studies(GWAS)within European ancestors and applied them as instrumental variable to GWAS of type 2 diabetes mellitus.After excluding SNPs that were significantly associated with smoking,physical activity,alcohol consumption,educational attainment,obesity,or type 2 diabetes mellitus,we assessed the impact of insomnia on type 2 diabetes mellitus using inverse variance weighting(IVW)method.Weighted median and MR-Egger regression analysis were also conducted to test the robustness of the association.We calculated the F statistic of the selected SNPs to test the applicability of instrumental variable and F statistic over than ten indicated that there was little possibility of bias of weak instrumental variables.We further examined the existence of pleiotropy by testing whether the intercept term in MR-Egger regression was significantly different from ze-ro.In addition,the leave-one-out method was used for sensitivity analysis to verify the stability and relia-bility of the results.Results:We selected 248 SNPs independently associated with insomnia at the genome-wide level(P＜5 ×10-8)as a preliminary candidate set of instrumental variables.After clum-ping based on the reference panel from 1000 Genome Project and removing the potential pleiotropic SNPs,a total of 167 SNPs associated with insomnia were included as final instrumental variables.The F statistic of this study was 39.74,which was in line with the relevance assumption of Mendelian randomi-zation.IVW method showed insomnia was associated with higher risk of type 2 diabetes mellitus that po-pulation with insomnia were 1.14 times more likely to develop type 2 diabetes mellitus than those without insomnia(95％CI:1.09-1.21,P＜0.001).The weighted median estimator(WME)method and MR-Egger regression showed similar causal effect of insomnia on type 2 diabetes mellitus.And MR-Egger re-gression also showed that the effect was less likely to be triggered by pleiotropy.Sensitivity analyses pro-duced directionally similar estimates.Conclusion:Insomnia is a risk factor of type 2 diabetes mellitus,which has positively effects on type 2 diabetes mellitus.Our study provides further rationale for indivi-duals at risk for diabetes to keep healthy lifestyle.

Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.

Objective:To evaluate the efficacy and safety of Xiao′er Huangjin Zhike Granules in the treatment of cough caused by acute bronchitis in children, which is defined in TCM terms as a syndrome of phlegm-heat obstructing the lung.Methods:This was a block-randomized, double-blind, placebo-controlled, multicenter clinical trial.From January 2022 to September 2023, 359 children aged 3 to 7 years old diagnosed as acute bronchitis (lung-obstructing phlegm-heat syndrome) were enrolled from 21 participating hospitals and randomly assigned to the experimental group and placebo group in a 3︰1 ratio, and respectively treated with Xiao′er Huangjin Zhike Granules and its matching placebo.Cough resolution/general resolution rate after 7 days of treatment was used as the primary efficacy outcome for both groups.Results:(1)On the seventh day of treatment, the rate of cough disappearance/basically disappearance in the experimental group and placebo group were 73.95% and 57.61% retrospectively, which had statistically significance ( P=0.001).(2)After 7 days of treatment, the median duration of cough disappearance/basic disappearance were 5 days and 6 days in the two groups , with a statistically significant difference ( P=0.006).The area under the curve of cough symptom severity time was 7.20 ± 3.79 in the experimental group and 8.20±4.42 in the placebo group.The difference between the two groups was statistically significant ( P=0.039).(3) After 7 days of treatment, the difference between TCM syndrome score and baseline was -16.0 (-20.0, -15.0) points in the experimental group and -15.0 (-18.0, -12.0) points in the placebo group, with significant difference between the two groups ( P=0.004).In the experimental group, the clinical control rate, the markedly effective rate, the effective rate and the ineffective rate were 49.04%, 28.35%, 16.48% and 6.13% severally; and in the placebo group, the clinical control rate, the markedly effective rate, the effective rate and the ineffective rate were 38.04%, 26.09%, 29.35%, and 6.52% separately, which had statistically significant ( P=0.014).(4) There was no significant difference in the incidence of adverse events or adverse reactions during the trial between both groups.Moreover, while adverse reactions in the form of vomiting and diarrhea were occasionally reported, no serious drug-related adverse event or adverse reaction was reported.(5)The tested drug provided good treatment compliance, showing no statistically significant difference from the placebo in terms of compliance rate. Conclusions:Based on the above findings, it can be concluded that Xiao′er Huangjin Zhike Granules provides good safety, efficacy, and treatment compliance in the treatment of cough caused by acute bronchitis, and lung-obstructing phlegm-heat syndrome, in children.

Purpose: This study aimed to identify the altered pathways and genes associated with freezing damage in human sperm during cryopreservation by multiomics analysis. Materials and Methods: Fifteen fresh human semen samples were collected for transcriptomic analysis, and another 5 fresh human semen samples were obtained for metabolomic analysis. For each semen sample, 1 mL was cryopreserved, and another 1 mL was left untreated for paired design. The results were then combined with previously published proteomic results to identify key genes/pathways. Results: Cryopreservation significantly reduced sperm motility and mitochondrial structure. Transcriptomic analysis revealed altered mitochondrial function, including changes in tRNA-methyltransferase activity and adenosine tri-phosphate/adenosine di-phosphate transmembrane transporter activity. Metabolomic analysis showed that the citrate cycle in mitochondria was significantly altered. Combining transcriptomic, proteomic, and metabolomic analyses revealed 346 genes that were altered in at least two omics analyses. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that metabolic pathways were significantly altered and strongly associated with mitochondria. Five genes were altered in all three omics analyses: COL11A1, COL18A1, LPCAT3, NME1, and NNT. Conclusions Five genes were identified by multiomics analysis in human cryopreserved sperm. These genes might have specific functions in cryopreservation. Explorations of the functions of these genes will be helpful for sperm cryopreservation and sperm motility improvement or even for reproduction in the future.

Objective To explore the effect of G-protein pathway suppressor 2(GPS2)on the proliferation and migration of HepG2 cells and the underlying mechanism.Methods GPS2 expression was analyzed via The Cancer Genome Atlas(TCGA)and Clinical Proteomic Tumor Analysis Consortium(CPTAC)online database.HepG2 cells with stable knockdown or overexpression of GPS2 were established with lentivirus.The protein and mRNA expression levels of GPS2 were detected by Western blotting and real-time quantitative PCR(qPCR)while cell proliferation was verified by cell proliferation assay.Cell migration was tested by Transwell and scratch assay.Epithelial-mesenchymal transition(EMT)biomarkers and the expression of matrix metalloproteinase(MMP)were detected by qPCR.Finally,the expressions of phosphorylation of protein kinase B(AKT)(p-AKT)and phosphorylation of extracellular signal-regulated kinase(ERK)(p-ERK)were detected by Western blotting.Results Based on the analysis of TCGA and CPTAC online database,GPS2 was highly expressed in human liver cancer tissues.Knockdown of GPS2 inhibited the proliferation and migration of HepG2 cells,while overexpression of GPS2 promoted the proliferation and migration of HepG2 cells.Silence of GPS2 up-regulated the mRNA level of E-cadherin(E-CAD),down-regulated the mRNA levels of N-cadherin(N-CAD),Vimentin(VIM),MMP2 and MMP9,and reduced the p-AKT and p-ERK.In contrast,overexpression of GPS2 decreased the mRNA level of E-CAD,increased the mRNA levels of N-CAD,VIM,MMP2 and MMP9,and elevated the protein levels of p-AKT and p-ERK.Conclusion GPS2 can promote the proliferation and migration of HepG2 cells,which might be attributed to increased activation of MAPK/ERK and PI3K/AKT signaling pathways and the EMT process.