Objective To exploring the drug rules for treating children's night cough based on the theory of"Shaoyang as the pivot".Methods 189 cases of children with night cough were included,and 224 prescriptions.Used the Traditional Chinese Medicine Inheritance Assistance Platform to analyze the time distribution,syndrome types,and four nature,five flavors,and channel tropism of diseases.Used frequency statistics,association rule analysis,and cluster analysis to extract drug patterns for pediatric night cough.Results Coughing occured most frequently during the Yin period.The syndrome type was mainly Shaoyang syndrome.The high-frequency core drugs were Chaihu,Huangqin,and Banxia,etc..The prescription characteristics were Xiaochaihu decoction without Renshen,Shengjiang,and Dazao,and added Wuweizi,Danggui and Xingren.The drugs were used flexibly according to the syndromes:Maxing Er San decoction was added to the phlegm and drink syndrome,Qumai Er Chen decoction was added to the food stagnation syndrome,Sijunzi decoction was added to the qi deficiency syndrome,Maiwei Dihuang decoction was added to the yin deficiency syndrome,and Weijing decoction was added to the phlegm and heat syndrome.Conclusion Based on the basic principle of harmonizing Shaoyang,and according to the disease mechanism,the classical prescription is flexibly used,forming a night cough treatment system based on the"Shaoyang as the pivot"theory,with distinct clinical characteristics.

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 14(LncRNA SNHG14)on high glucose-induced podocyte injury by targeting microRNA-30a-5p(miR-30a-5p).Methods Podocytes were cultured in vitro and were divided into the following groups:the standard glucose(NG)group,the high glucose(HG)group,the si-NC+HG group,the si-SNHG14+HG group,the miR-NC+HG group,the miR-30a-5p mimics+HG group,the si-SNHG14+inhibitor NC+HG group and the si-SNHG14+miR-30a-5p inhibitor+HG group.Quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect expression levels of LncRNA SNHG14 and miR-30a-5p.Flow cytometry was used to determine cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-6 and IL-1β.Xanthine oxidase method,ammonium molybdate colorimetry and thiobarbituric acid method were respectively used to detect superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA).Dual-luciferase reporter gene was conducted to verify the targeting relationship between LncRNA SNHG14 and miR-30a-5p.Western blot assay was performed to detect expression levels of apoptosis-related proteins.Results Compared with the NG group,the HG group exhibited increased expression levels of LncRNA SNHG14,cell apoptosis rate,as well as levels of TNF-α,IL-6,IL-1β and MDA,whereas the expression level of miR-30a-5p and levels of SOD and CAT were decreased(P＜0.05).Compared with the HG group and the si-NC+HG group,the si-SNHG14+HG group exhibited decreased expression levels of LncRNA SNHG14,apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins,while the expression level of miR-30a-5p,levels of SOD and CAT and the expression level of Bcl-2 protein were increased(P＜0.05).Compared with the HG group and the miR-NC+HG group,the miR-30a-5p mimics+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05).Meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were increased,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were decreased(P＜0.05).Compared with the si-SNHG14+HG group and the si-SNHG14+inhibitor NC+HG group,the si-SNHG14+miR-30a-5p inhibitor+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05),meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were reduced,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were increased(P＜0.05).The dual-luciferase reporter gene assay confirmed that LncRNA SNHG14 targeted and negatively regulated miR-30a-5p.Conclusion The inhibition of LncRNA SNHG14 can target miR-30a-5p to alleviate high glucose induced podocyte injury.

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 14(LncRNA SNHG14)on high glucose-induced podocyte injury by targeting microRNA-30a-5p(miR-30a-5p).Methods Podocytes were cultured in vitro and were divided into the following groups:the standard glucose(NG)group,the high glucose(HG)group,the si-NC+HG group,the si-SNHG14+HG group,the miR-NC+HG group,the miR-30a-5p mimics+HG group,the si-SNHG14+inhibitor NC+HG group and the si-SNHG14+miR-30a-5p inhibitor+HG group.Quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect expression levels of LncRNA SNHG14 and miR-30a-5p.Flow cytometry was used to determine cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-6 and IL-1β.Xanthine oxidase method,ammonium molybdate colorimetry and thiobarbituric acid method were respectively used to detect superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA).Dual-luciferase reporter gene was conducted to verify the targeting relationship between LncRNA SNHG14 and miR-30a-5p.Western blot assay was performed to detect expression levels of apoptosis-related proteins.Results Compared with the NG group,the HG group exhibited increased expression levels of LncRNA SNHG14,cell apoptosis rate,as well as levels of TNF-α,IL-6,IL-1β and MDA,whereas the expression level of miR-30a-5p and levels of SOD and CAT were decreased(P＜0.05).Compared with the HG group and the si-NC+HG group,the si-SNHG14+HG group exhibited decreased expression levels of LncRNA SNHG14,apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins,while the expression level of miR-30a-5p,levels of SOD and CAT and the expression level of Bcl-2 protein were increased(P＜0.05).Compared with the HG group and the miR-NC+HG group,the miR-30a-5p mimics+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05).Meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were increased,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were decreased(P＜0.05).Compared with the si-SNHG14+HG group and the si-SNHG14+inhibitor NC+HG group,the si-SNHG14+miR-30a-5p inhibitor+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05),meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were reduced,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were increased(P＜0.05).The dual-luciferase reporter gene assay confirmed that LncRNA SNHG14 targeted and negatively regulated miR-30a-5p.Conclusion The inhibition of LncRNA SNHG14 can target miR-30a-5p to alleviate high glucose induced podocyte injury.

Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.

Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.

To report a case of an immunocompetent young adult male patient diagnosed with intracranial Aspergillus flavus infection， and to investigate the clinical features of this disease and related experience in diagnosis and treatment.A retrospective analysis was performed for the clinical data of a patient who had the initial presentation of high fever and headache and then progressed to meningoencephalitis， and the results of cerebrospinal fluid （CSF） metagenomic next-generation sequencing （mNGS） and treatment outcomes were summarized.The patient had an acute onset， with no response to empirical anti-infective therapy in the incipient stage， and then he gradually developed disturbance of consciousness and meningeal irritation sign. CSF analysis showed inflammatory changes， while conventional pathogen tests yielded negative results， and mNGS detected 27 specific sequences of Aspergillus flavus. The symptoms of the patient was significantly improved after antifungal therapy with voriconazole， with no recurrence after follow-up for 3 months.For unexplained central nervous system infections， especially those with negative results from conventional tests， mNGS can improve the detection rate of rare pathogens（e.g.，Aspergillus flavus）. Early diagnosis and targeted antifungal therapy are crucial for improving prognosis. This case highlights that invasive fungal infections should be considered even in immunocompetent individuals.
Aspergillus flavus

OBJECTIVE: While higher therapeutic doses of toxic cardiac glycosides derived from Cerbera odollam are frequently employed in cases of suicide or homicide, ongoing research is investigating the potential anticancer properties of low-concentration extracts obtained from the fruits of C. odollam. The present study aimed to determine the enhanced anticancer effects and minimize potential side effects of combining extracts from C. odollam fruits from Thailand with sorafenib against HCT116 and HepG2 cells. METHODS: The dried powder of fresh green fruits of C. odollam was fractionated, and its phytochemical contents, including total cardiac glycosides, phenolics, flavonoids, and triterpenoids, were quantified. The cytotoxic effects of these fractions were evaluated against HCT116 and HepG2 cells using the MTT assay. The fractions showing the most significant response in HCT116 and HepG2 cells were subsequently combined with sorafenib to examine their synergistic effects. Apoptosis induction, cell cycle progression, and mitochondrial membrane potential (MMP) were then assessed. The underlying mechanism of the apoptotic effect was further investigated by analyzing reactive oxygen species (ROS) generation and the expression levels of antioxidant proteins. RESULTS: Phytochemical analysis showed that C. odollam-ethyl acetate fraction (COEtOAc) was rich in cardiac glycosides, phenolics, and flavonoids, while the dichloromethane fraction (CODCM) contained high levels of triterpenoids and saponins. Following 24 h treatment, HCT116 showed the most significant response to COEtOAc, while HepG2 responded well to CODCM with IC₅₀ values of (42.04 ± 16.94) μg/mL and (123.75 ± 14.21) μg/mL, respectively. Consequently, COEtOAc (20 μg/mL) or CODCM (30 μg/mL), both administered at sub-IC₅₀ concentrations, were combined with sorafenib at 6 μmol/L for HCT116 cells and 2 μmol/L for HepG2 cells, incubated for 24 h. This combination resulted in a significant suppression in cell viability by approximately 50%. The combination of treatments markedly enhanced apoptosis, diminished MMP, and triggered G₀/G₁ phase cell cycle arrest compared to the effects of each treatment administered individually. Concurrently, increased formation of ROS and decreased expression of the antioxidant enzymes superoxide dismutase 2 and catalase supported the proposed mechanism of apoptosis induction by the combination treatment. Importantly, the anticancer effect demonstrated a specific targeted action with a favorable safety profile, as evidenced by HFF-1 cells displaying IC₅₀ values 2-3 times higher than those of the cancer cells. CONCLUSION Utilizing sub-IC₅₀ concentrations of COEtOAc or CODCM in combination with sorafenib can enhance targeted anticancer effects beyond those achieved with single-agent treatments, while mitigating opposing side effects. Future research will focus on extracting and characterizing active constituents, especially cardiac glycosides, to enhance the therapeutic potential of anticancer compounds derived from toxic plants.

Objective:To investigate the diagnostic value of plasma cytokines such as interleukin (IL)-2, IL-6 and interferon (IFN)-γ in non-Hodgkin lymphoma (NHL).Methods:A retrospective case-control study was conducted. A total of 48 NHL patients admitted to the Second Affiliated Hospital of Xuzhou Medical University between January 2020 and December 2022 were selected as NHL group, and another 34 healthy people who underwent physical examimation during the same period were selected as the healthy control group. The levels of IL-2, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor (TNF) - α and IFN-γ in the plasma of patients at first admission and healthy subjects during physical examination were detected by using flow cytometry. The differences in general data and all cytokines levels of both groups were compared. The collinearity stepwise screening was made in 7 cytokines levels, and the screened variables were included in multivariate binary logistic regression model. Plasma cytokines with independent effects on the pathogenesis of NHL were screened. Taking local biopsy, histopathological examination or immunohistochemical examination as the gold standard, the receiver operating characteristic (ROC) curves of individual and combined diagnosis of NHL based on the selected cytokines were drawn to judge the diagnostic effect of all indicators on NHL.Results:There were 32 males (66.7%) and 16 females (33.3%) in NHL group, with the median age [ M ( Q1, Q3)] of 56.50 (45.75, 67.50) years; there were 28 males (82.4%) and 6 females (17.6%) in the healthy control group, with the median age of 52.00 (47.50, 55.50) years. There were no statistically significant differences in age and gender composition between the 2 groups (all P > 0.05). The levels of IL-2 [1.44 (1.36, 1.85) pg/ml vs. 1.19 (0.86, 1.68) pg/ml] and TNF-α [3.46 (2.68, 4.06) pg/ml vs. 2.23 (1.52, 3.46) pg/ml] in NHL group were higher than those in the healthy control group, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in IL-4, IL-6, IL-10, IL-17, IFN-γ levels (all P > 0.05). According to collinear stepwise screening of independent variables, IL-4 and TNF-α were excluded from 7 cytokines, and the other 5 cytokines were included in multivariate logistic regression model, and the result showed that the decreased level of IL-2 ( OR = 0.20, 95% CI: 0.08-0.53, P = 0.001) and the increased levels of IL-6 ( OR = 1.18, 95% CI: 1.04-1.33, P = 0.009) and IFN-γ ( OR = 1.26, 95% CI: 1.08-1.46, P = 0.003) were independent risk factors for the onset of NHL. The results showed that the area under the curve of IL-2, IL-6, IFN-γ and the combination of 3 indexes for the diagnosis of NHL was 0.760 (95% CI: 0.651-0.870), 0.595 (95% CI: 0.468-0.722), 0.508 (95% CI: 0.373-0.642), 0.847 (95% CI: 0.763-0.930), and the optimal cut-off value of the combination of 3 indexes was 0.730 which was calculated by logistic regression model formula; the corresponding sensitivity and specificity were 70.2% and 94.1%, respectively. Conclusions:The decreased level of IL-2 and increased levels of IL-6 and IFN-γ at initial diagnosis are risk factors for the onset of NHL. The combined detection of the 3 indexes shows a good value in the diagnosis of NHL.

Objective: To explore the influencing factors for sarcopenia among elderly male patients with type 2 diabetes (T2DM), so as to provide the basis for the early prevention and treatment of sarcopenia. Methods: Male T2DM patients aged 60 and above admitted to Hangzhou First People's Hospital from January to December 2024 were selected as the study subjects. Demographic data, T2DM complications, and blood biochemical parameters were collected. Physical activity levels were assessed using the Chinese version of the International Physical Activity Questionnaire. The diagnosis of sarcopenia was made according to the diagnostic procedures and criteria established by the Asian Working Group for Sarcopenia in 2019. Factors affecting sarcopenia among elderly male patients with T2DM were analyzed using multivariable logistic regression model. Results: A total of 455 elderly male patients with T2DM were surveyed, with a mean age of (71.80±9.55) years. The predominant physical activity level was moderate with 226 cases accounting for 49.67%. The disease course of T2DM was mainly from 10-<20 years, with 229 cases accounting for 50.33%. There were 140 cases of T2DM complications, accounting for 30.77%. A total of 138 cases of sarcopenia were detected, with a prevalence of 30.33%. Multivariable logistic regression analysis showed that age (OR=1.077, 95%CI: 1.003~1.156), body mass index (<18.5kg/m2, OR=11.056, 95%CI: 3.343~36.547; 18.5~<25.0 kg/m2, OR=2.633, 95%CI: 1.420~4.881), physical activity level (low, OR=2.469, 95%CI: 1.421~4.292), chronic obstructive pulmonary disease (yes, OR=1.871, 95%CI: 1.091~3.206), T2DM complications (yes, OR=3.015, 95%CI: 1.516~6.001), glycated hemoglobin (≥7%, OR=2.822, 95%CI: 1.423~5.590) and albumin (OR=0.810, 95%CI: 0.662~0.991) were factors affecting sarcopenia among elderly male patients with T2DM (P<0.05). Conclusion Advanced age, body mass index <25.0 kg/m2, low physical activity level, chronic obstructive pulmonary disease, T2DM complications, high glycated hemoglobin and low albumin are associated with a higher risk of sarcopenia in elderly male patients with T2DM.

Objective:To investigate the role of mechanosensor Yes-associated protein 1 (YAP1) in urothelial cells in inducing bladder dysfunction in a partial bladder outlet obstruction (pBOO) model.Methods:Ten female C57BL/6 mice were included in this study and randomly divided into pBOO and sham groups based on body weight using a stratified pairing method, with 5 mice in each group. The pBOO group underwent proximal urethral ligation surgery, while the sham group underwent a sham operation. Two weeks after surgery, the urinary pattern was analyzed using the urine spot test. The significant increase in urine spot numbers indicated the successful establishment of the pBOO model. The mice were then sacrificed, and bladder tissues were weighed and stained with hematoxylin and eosin (HE) to observe morphological changes. The bladder urothelial layer was further isolated, and total cell proteins were extracted to detect the expression levels of YAP1 protein using Western blotting. Mouse immortalized bladder urothelial cells were divided into three experimental groups: the negative control (NC) group, which was treated with YAP1-NC lentivirus; the overexpression (OE) group, which was treated with YAP1-OE lentivirus to induce YAP1 protein overexpression; and the verteporfin treatment (VP) group, which was treated with verteporfin on the basis of the OE group. Real-time quantitative PCR and Western blotting were used to verify the transcription and expression levels of YAP1 protein, the co-transcriptional activator TEAD4 protein, and the phosphorylated protein DRP1-616 (at serine 616) of dynamin-related protein 1 (DRP1). An ATP detection kit was used to measure the ATP release concentration in the NC, OE, and VP groups. The interaction between YAP1 and TEAD4 was investigated using co-immunoprecipitation, and the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 20 (Tom20) was observed using immunofluorescence staining.Results:The results of the urine spot test showed that the number of urine spots on the filter paper in the pBOO group was higher than that in the sham group within 6 hours [(283.0±9.1) spots vs. (3.7±0.3) spots, P<0.01], and the urine spots were scattered. The bladder wet weight in the pBOO group was significantly higher than that in the sham group [(105.70±6.84) mg vs. (22.33±1.20) mg, P<0.01]. Histological observations revealed reduced bladder mucosal folds and increased detrusor muscle thickness in the pBOO group. The expression of YAP1 protein in the bladder urothelial cells of the pBOO group was significantly upregulated compared to the sham group [(1.26±0.08) vs. (0.50±0.04), P<0.01]. In vitro experiments showed that compared to the NC group, the OE group had significantly increased expression of DRP1-616 [(0.94±0.05) vs. (0.33±0.01), P<0.01] and higher ATP release concentration [(24.45±0.16) μmol/mg vs. (19.67±0.42) μmol/mg, P<0.01]. In contrast, the VP group had significantly decreased expression of DRP1-616 [(0.29±0.04) vs. (0.94±0.05), P<0.01] and lower ATP release concentration [(10.55±0.01) μmol/mg vs. (24.45±0.16) μmol/mg, P<0.01] compared to the OE group. Co-immunoprecipitation experiments using YAP1 and TEAD4 antibodies showed that YAP1 and TEAD4 proteins could interact and form a transcriptional complex to regulate ATP release. Immunofluorescence staining revealed increased expression of Tom20 in the OE group compared to the NC group [(104.20±3.28) vs. (74.51±3.87), P<0.01]. Conclusions:In the pBOO-induced bladder dysfunction model, YAP1 is highly expressed in urothelial cells. YAP1 forms a transcriptional complex with TEAD4 to regulate ATP release by promoting mitochondrial fission via DRP1-616 expression, which is a key mechanism underlying pBOO-induced bladder dysfunction.