Objective To analyze the detrimental effects of exposure to environmental noise alone and combined with carbon monoxide(CO)on spermatogenesis in male rats,investigate the underlying mechanisms involved in such damage,and evaluate the protective role of pterostilbene(PTE)against these adverse effects.Methods Sixty male SD rats(6～8 weeks old,weighing 200±10 g)were randomly divided normal control group(standard housing),sham-exposure control group(restraint stress only),noise exposure(85 dB),CO exposure(460 mg/m3),combined exposure(simultaneous exposure),and PTE intervention(80 mg/kg pretreatment),with 10 animals in each group.The rats were exposed daily for 2 h via a nose-only inhalation exposure system within a multifactorial environmental simulation chamber for 60 consecutive days.Sperm count and viability were measured after exposure.Histopathological changes of testicular tissues were observed with HE staining.qRT-PCR was used to measure stage-specific mRNA levels in germ cells.Serum sex hormone levels and adenosine triphosphate(ATP)concentrations in testes and sperm were detected.Transmission electron microscopy(TEM)was applied to observe the ultrastructural damage in the spermatocytes.Additionally,transcriptome sequencing was performed on testicular tissue,followed by bioinformatics analysis.Results Compared with the negative control group,the combined exposure group exhibited significant reductions in sperm viability and count(P<0.05),and developmental arrest of immature germ cells in the testicular tissue,with obviously less spermatogonia,spermatocytes,and round/elongated spermatids(P<0.01).Additionally,significantly reduced levels of reproductive-related hormones,such as gonadotropin-releasing hormone,follicle-stimulating hormone,luteinizing hormone and testosterone,and ATP levels in testes and sperm were observed in the mice after combined exposure(P<0.01),accompanied by mitochondrial rupture and cristae disruption in spermatocytes.Conversely,the PTE intervention group showed marked alleviation of these impairments,with parameters recovering almost to normal levels.Transcriptome sequencing identified biological processes related to reproductive development and ATP-dependent pathways as potential contributors to testicular injury induced by noise and CO exposure,with key genes including Nppa,Adm,Gnrh1,Ptafr,Atp13a5,Atp8b1,and LOC102555469.Conclusion Noise and CO exposure induce spermatogenic damage in rats,which may be related with energy metabolism and hormonal regulation,while PTE demonstrates significant protective effects against such reproductive impairments.

Objective To explore the impact of integrated traditional Chinese and Western medicine on the overall survival(OS)of patients with biliary tract carcinoma(BTC),and to analyze the related factors affecting the OS of patients with BTC,so as to provide accurate prognosis assessment for clinical diagnosis and treatment.Methods The medical records of 198 patients with BTC in Henan Provincial Hospital of Traditional Chinese Medicine from February 2018 to October 2023 were retrospectively analyzed.The Cox proportional hazards model was utilized to analyze the independent prognostic factors affecting the OS of BTC patients.Survival curves were constructed using the Kaplan-Meier method,and the Log-rank test was employed to compare median OS differences among various groups.Results A total of 116 patients with BTC were included.There were 59 cases(50.86％)in the Western medicine group,with a median OS of(14±2.77)months,and the 1-,2-and 3-year OS rates were 81.36％,61.02％and 46.11％,respectively.There were 57 cases(49.14％)in the integrated traditional Chinese and Western medicine group,with a median OS of(29±3.91)months,and the 1-,2-and 3-year OS rates were 82.32％,67.64％and 58.00％,respectively.There was a statistically significant difference in survival time between the two groups(P=0.038).Univariate analysis indicated that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy and integrated traditional Chinese and Western medicine were significantly correlated with OS in BTC patients(all P＜0.05).Multivariate analysis revealed that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine had significant effects on the OS of patients with BTC(all P＜0.05).Conclusion The median OS of the traditional Chinese and Western medicine groups was significantly longer than that of the Western medicine group,and the long-term OS rate of the patients was significantly better than that of the Western medicine group.Age,clinical stage,surgery,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine are independent prognostic factors affecting OS in patients with BTC.

AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P＜0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P＜0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P＜0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P＜0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P＞0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.

AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P＜0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P＜0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P＜0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P＜0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P＞0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.

Objective To explore the impact of integrated traditional Chinese and Western medicine on the overall survival(OS)of patients with biliary tract carcinoma(BTC),and to analyze the related factors affecting the OS of patients with BTC,so as to provide accurate prognosis assessment for clinical diagnosis and treatment.Methods The medical records of 198 patients with BTC in Henan Provincial Hospital of Traditional Chinese Medicine from February 2018 to October 2023 were retrospectively analyzed.The Cox proportional hazards model was utilized to analyze the independent prognostic factors affecting the OS of BTC patients.Survival curves were constructed using the Kaplan-Meier method,and the Log-rank test was employed to compare median OS differences among various groups.Results A total of 116 patients with BTC were included.There were 59 cases(50.86％)in the Western medicine group,with a median OS of(14±2.77)months,and the 1-,2-and 3-year OS rates were 81.36％,61.02％and 46.11％,respectively.There were 57 cases(49.14％)in the integrated traditional Chinese and Western medicine group,with a median OS of(29±3.91)months,and the 1-,2-and 3-year OS rates were 82.32％,67.64％and 58.00％,respectively.There was a statistically significant difference in survival time between the two groups(P=0.038).Univariate analysis indicated that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy and integrated traditional Chinese and Western medicine were significantly correlated with OS in BTC patients(all P＜0.05).Multivariate analysis revealed that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine had significant effects on the OS of patients with BTC(all P＜0.05).Conclusion The median OS of the traditional Chinese and Western medicine groups was significantly longer than that of the Western medicine group,and the long-term OS rate of the patients was significantly better than that of the Western medicine group.Age,clinical stage,surgery,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine are independent prognostic factors affecting OS in patients with BTC.

【Objective】 To explore the role of ZFP36 in cardiomyocyte injury and autophagy induced by hypoxia/reoxygenation (H/R) so as to clarify its molecular regulatory mechanism. 【Methods】 H9C2 rat cardiomyocytes were infected with ZFP36 overexpressing lentivirus (OE-ZFP36) or its negative control lentivirus (OE-ZFP36 NC) to construct stable cell lines, respectively. Transfection of ATG4D overexpression plasmid (OE-ATG4D) improved the expression of ATG4D. Hypoxia/reoxygenation (H/R) induced myocardial cell injury. H9C2 cells were mainly divided into control group, H/R group, OE-ZFP36 NC+H/R group, OE-ZFP36+H/R group, OE-ATG4D NC+H/R group, OE-ATG4D+H/R group, OE-ZFP36+OE-ATG4D NC+H/R group, and OE-ZFP36+OE-ATG4D+H/R group. The protein expressions of ATG4D, Beclin1, LC3 and ZFP36 in H9C2 cells were detected by Western blotting. The mRNA levels of ZFP36 and ATG4D in H9C2 cells were detected by Real-time fluorescence quantitative PCR (qPCR). The viability of H9C2 cells was detected by CCK-8 assay. The levels of interleukin (IL-6) and tumor necrosis factor (TNF-α) in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) in H9C2 cells were detected by DCFH-DA method. SOD detection kit was used to detect the SOD level in H9C2 cells. The apoptosis of H9C2 cells was detected by flow cytometry. LC3 autophagosomes in H9C2 cells were detected by cellular immunofluorescence. Dual-luciferase reporter gene assay was used to detect the binding of ZFP36 and ATG4D mRNA in H9C2 cells. 【Results】 Compared with control group, H/R group showed decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels and decreased SOD levels, increased cell apoptosis. Up-regulated ATG4D and Beclin1 protein expression, increased LC3Ⅱ/LC3Ⅰ ratio, as well as upregulated ZFP36 expression were found in H/R group (all P<0.05). Compared with OE-ZFP36 NC+H/R group, elevated cell viability, decreased IL-6 and TNF-α levels, decreased ROS levels and increased SOD levels, reduced cell apoptosis (P<0.05), and downregulated ATG4D and Beclin1 protein expression, decreased LC3Ⅱ/LC3Ⅰ ratio were shown in OE-ZFP36+H/R group (all P<0.05). Compared with infection with OE-ZFP36 NC lentivirus, infection with OE-ZFP36 lentivirus decreased the luciferase activity of ATG4D 3′-UTR reporter gene, decreased the stability of ATG4D mRNA, and downregulated the H/R-induced ATG4D mRNA expression (all P<0.05). Compared with OE-ATG4D NC+H/R group, OE-ATG4D+H/R group had upregulated ATG4D mRNA and protein expression, decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). Compared with OE-ZFP36+OE-ATG4D NC+H/R group, OE-ZFP36+OE-ATG4D+H/R group had decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). 【Conclusion】 The expression of ZFP36 is upregulated in H/R-induced cardiomyocyte injury. The overexpression of ZFP36 inhibits H/R-induced cardiomyocyte injury and autophagy by regulating ATG4D, thus resisting cardiomyocyte H/R injury. It proves that ZFP36 is an important regulatory molecule against MI/RI.

Objective Based on the endoplasmic reticulum stress(ERS)inositol-requiring enzyme-1α(IRE1α)/apoptosis signal-regulated kinase 1(ASK1)/c-Jun N-terminal kinase(JNK)pathway to investigate the intervention effect of Ziziphi Spinosae Semen(ZSS)-Albiziae Flos(AF)on the depression model of rats,which were prepared by solitary rearing combined with chronic unpredictable mild stress(CUMS).Methods A total of 144 rats were randomly divided into normal group,model group,high-,medium-and low-dose groups of ZSS-AF,and Venlafaxine group.In addition to the normal group,the rats in other groups were isolated for 21 days combined with CUMS to prepare the depression model.The behavioral changes of rats were observed by open field test and Morris water maze test.The ultrastructural changes of hippocampal neurons were observed by transmission electron microscope.TUNEL staining was used to observe the apoptosis of nerve cells in hippocampus.The protein expression levels of IRE1α,phosphorylated(P)-IRE1α,ASK1,P-ASK1,JNK,P-JNK,B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteme aspartate specific protease-3(Caspase-3)in hippocampus were detected by Western Blot.The mRNA expression levels of IRE1α,ASK1,JNK,Bax,Bcl-2 and Caspase-3 in hippocampus were detected by Real-Time PCR.Results Compared with the normal group,the scores of horizontal movement and vertical movement in the open field test of rats in the model group were decreased(P＜0.01).In the water maze test,the escape latency was increased and the number of crossing the original platform was decreased(P＜0.01).The number of hippocampal apoptosis was increased(P＜0.01).The protein expression levels of P-IRE1 α/IRE1 α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 and mRNA expressions of IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were increased,while the protein and mRNA expression levels of Bcl-2 were decreased(P＜0.01).Compared with model group,the scores of horizontal movement and vertical movement in the open field test of rats in each dose ZSS-AF groups and Venlafaxine group were increased(P＜0.01).In the water maze test,the escape latency was decreased and the times of crossing the original platform was increased(P＜0.01).The number of hippocampal apoptosis was decreased(P＜0.01).The mRNA expression levels of P-IRE1α/IRE1α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 protein and IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were decreased,while the protein and mRNA expression levels of Bcl-2 were increased(P＜0.05,P＜0.01).The effect of medium-dose ZSS-AF group was better than that of high-and low-dose groups.Conclusion ZSS-AF may play an antidepressant role by regulating IRE1α/ASK1/JNK pathway of endoplasmic reticulum stress.

Objective To explore the clinical application value of emergency bedside echocardiography(EB-Echo)in the diagnosis and treatment of reverse-Takotsubo syndrome(r-TTS).Methods A retrospective study method was conducted,the EB-Echo imaging and clinical data of 10 patients already diagnosed with r-TTS and admitted to the department of critical care medicine of Zhengzhou People's Hospital from January 2014 to December 2021 were selected.The r-TTS group,while data from 33 concurrently admitted patients with classic Takotsubo syndrome(TTS)were selected as the classic TTS group.Comparisons were made in terms of EB-Echo detection rates,diagnosis times,cardiac function indicators[left ventricular ejection fraction(LVEF),left ventricular stroke volume(LVSV)],blood volume indicators[inferior vena cava diameter(IVCD),collapse degree of inferior vena cava(cIVC)],complications[acute left ventricular outflow tract obstruction(ALVOTO),acute mitral regurgitation(AMR),and pulmonary hypertension(PH)],and prognosis between the two groups.Results The detection rate of EB-Echo in the r-TTS group was significantly higher than that in the classic TTS group[100.0%(10/10)vs.66.7%(22/33),P<0.05],and the diagnosis time was significantly shorter(minutes:26.80±3.77 vs.41.18±6.61,P<0.05).In terms of cardiac function indicators,both LVEF and LVSV were significantly lower in the r-TTS group compared to the classic TTS group[LVEF:0.36±0.05 vs.0.41±0.04,LVSV(mL):36.43±4.30 vs.40.65±5.09,both P<0.05].However,there were no significant differences in blood volume indicators(IVCD and cIVC)between the r-TTS group and the classic TTS group[IVCD(mm):15.02±1.88 vs.14.94±1.75,cIVC:(0.43±0.06)%vs.(0.44±0.07)%,both P>0.05].Concerning complications,the incidence of ALVOTO was slightly lower in the r-TTS group compared to the classic TTS group[0%(0/10)vs.21.2%(7/33),P>0.05],while the incidences of AMR and PH were similar between the two groups[AMR incidence:40.0%(4/10)vs.24.2%(8/33),PH incidence:30.0%(3/10)vs.18.2%(6/33),both P>0.05].After a follow-up ranging from 6 months to 3 years,no deaths were reported in the r-TTS group,whereas there was one fatality in the classic TTS group.Nevertheless,the mortality difference between the groups was not statistically significant.Conclusion EB-Echo demonstrates significant clinical value in the early diagnosis,guiding therapy,and enhancing prognosis of r-TTS.

Objective:To study the relationship between the symptoms and medication law of fever treated with modified Zhuye Shigao Decoction by means of data mining.Methods:Clinical medical records of modified Zhuye Shigao Decoction for the treatment of fever were retrieved from CNKI, VIP, Wanfang Data, CBM from the establishment of the databases to October 31, 2022. Excel 2019 was used to conduct statistics on symptoms and drug frequency. SPSS Modeler 18.0 and SPSS Statistics 25.0 were used to build complex networks and system clusters, and R 4.2.1 was used to analyze the association rules of the database. Results:A total of 163 medical records were included, involving 143 symptoms, mainly fever, anorexia, thirst, upset, sweating, etc; 40 tongue and pulse conditions, mainly including pulse number and red tongue; 163 kinds of Chinese materia medica were involved, including 9 main kinds of Chinese materia medica. The frequency and complex network of Chinese materia medica indicated that the main medicines for fever were the original prescriptions of Zhuye Shigao Decoction, and the commonly used doses were Folium Phyllostachydis Henonis 10 g, Gypsum Fibrosum 30 g, Ginseng Radix et Rhizoma or Codonopsis Radix 10 g, Ophiopogonis Radix 15 g, Pinelliae Rhizoma 10 g, Glycyrrhizae Radix et Rhizoma 6 g, and Oryzae Semen 15g; According to the clustering analysis of compatibility drugs, five core compatibility combinations were obtained. "Forsythiae Fructus→Lonicerae Japonicae Flos", "Adenophorae Radix→dry mouth" had strong correlation.Conclusion:The syndrome types of fever treated by Zhuye Shigao Decoction were unclear exogenous heat-qi and yin injury, phlegm and heat obstructing the lung, excessive heat in the interior-fluid deficiency and heat stagnation, stomach yin deficiency and fire hyperactivity, spleen and stomach weakness-edema due to yang deficiency; the cause of fever treated by Zhuye Shigao Decoction could originate from any of the three energizers, and the disease location was mainly in the stomach.

BACKGROUND: Osteopenia has been well documented in adolescent idiopathic scoliosis (AIS). Bone marrow stem cells (BMSCs) are a crucial regulator of bone homeostasis. Our previous study revealed a decreased osteogenic ability of BMSCs in AIS-related osteopenia, but the underlying mechanism of this phenomenon remains unclear. METHODS: A total of 22 AIS patients and 18 age-matched controls were recruited for this study. Anthropometry and bone mass were measured in all participants. Bone marrow blood was collected for BMSC isolation and culture. Osteogenic and adipogenic induction were performed to observe the differences in the differentiation of BMSCs between the AIS-related osteopenia group and the control group. Furthermore, a total RNA was extracted from isolated BMSCs to perform RNA sequencing and subsequent analysis. RESULTS: A lower osteogenic capacity and increased adipogenic capacity of BMSCs in AIS-related osteopenia were revealed. Differences in mRNA expression levels between the AIS-related osteopenia group and the control group were identified, including differences in the expression of LRRC17 , DCLK1 , PCDH7 , TSPAN5 , NHSL2 , and CPT1B . Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed several biological processes involved in the regulation of autophagy and mitophagy. The Western blotting results of autophagy markers in BMSCs suggested impaired autophagic activity in BMSCs in the AIS-related osteopenia group. CONCLUSION Our study revealed that BMSCs from AIS-related osteopenia patients have lower autophagic activity, which may be related to the lower osteogenic capacity and higher adipogenic capacity of BMSCs and consequently lead to the lower bone mass in AIS patients.
Humans ; Adolescent ; Scoliosis/genetics* ; Cell Differentiation/physiology* ; Osteogenesis/genetics* ; Bone Diseases, Metabolic/genetics* ; Kyphosis ; Autophagy/genetics* ; Bone Marrow Cells ; Cells, Cultured ; Doublecortin-Like Kinases