ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

BACKGROUND:Research suggests that exercise interventions may be more advantageous than pharmacologic treatments or dietary restriction alone for fat loss,but fewer studies have simultaneously examined the effects of training modalities and training cycles on visceral and subcutaneous fat in obese populations. OBJECTIVE:To investigate the impact of training modalities and training cycles on visceral and subcutaneous fat in recessive obesity female college students. METHODS:Sixty-three female college students with hidden obesity(body mass index<24 kg/m2 and body fat content percentage≥30%)were recruited from Zhengzhou College of Commerce and Industry,and were randomly divided into a high-intensity intermittent training group(n=32)and a moderate-intensity continuous training group(n=31)using the lottery method.Subjects in both groups performed exercise training of corresponding intensity on a running platform and ensured that the exercise volume of both groups was equal,3 times per week,every 4 weeks as one training cycle for 16 weeks.Before training and at the end of each training cycle,subjects'visceral fat,subcutaneous fat,and overall fat were measured using the corresponding test devices. RESULTS AND CONCLUSION:The repeated measures F results indicated that the main effects of training cycles on visceral fat area,visceral fat index,abdominal subcutaneous fat thickness,percentage of body fat and body mass index were significant,while the main effects of training modalities were significant for subcutaneous fat thickness in the triceps brachii and scapula regions.All the interaction effects between training modalities and training cycles were significant(P<0.05).Results from the simple effect tests revealed that the significant simple effects of training modalities at the 4th and 12th weeks for visceral fat area and visceral fat index,at the 8th and 12th weeks for subcutaneous fat thickness in the triceps brachii,scapula region,and abdominal regions,and at the 8th week for the percentage of body fat and body mass index.Simple effects of training cycles were significant for all measures within each group.(3)The results of multiple comparison tests indicated that in the high-intensity intermittent training group,visceral fat area,visceral fat index,percentage of body fat,body mass index and abdominal subcutaneous fat thickness decreased sequentially at the 4th,8th,12th,and 16th weeks,and subcutaneous fat thickness of the triceps brachii and scapula decreased sequentially at the 8th,12th,and 16th weeks(P<0.05).In the moderate-intensity continuous training group,visceral fat area,visceral fat index,subcutaneous fat thickness of the triceps brachii and scapula,percentage of body fat and body mass index decreased successively at the 8th,12th,and 16th weeks,while abdominal subcutaneous fat thickness decreased successively at the 4th,8th,12th,and 16th weeks(P<0.05).To conclude,both training modalities and training cycles are factors influencing visceral and subcutaneous fat in recessive obesity female college students.Training modality primarily affects subcutaneous fat in the triceps brachii and scapular region,but the fat loss effects may converge over a longer training cycle;training cycle primarily affects visceral fat area,visceral fat index,abdominal subcutaneous fat,body fat content,and body mass index.

Objective To explore the correlation of MET status in patients with advanced prostatic acinar adenocarcinoma with the clinical pathological parameters and prognosis. Methods The specimen from 135 patients with advanced prostatic acinar adenocarcinoma was included. The expression of c-MET protein was detected via immunohistochemistry, and MET gene amplification was assessed by fluorescence in situ hybridization. The relationships of c-MET expression and gene amplification with clinicopathological features and prognosis were analyzed. Results The positive expression rate of c-MET was 52.60% (71/135). Compared with the c-MET expression in adjacent tissues, that in tumor tissues showed lower heterogeneous expression. Among the cases, 1.71% (2/117) exhibited MET gene polyploidy, but no gene amplification was detected. Positive c-MET expression was significantly correlated with high Gleason scores and grade groups (P=0.0073). However, no significant relationship was found between c-MET expression and progression-free survival or post-treatment t-PSA levels. Conclusion c-MET protein is expressed at a relatively low level in advanced prostatic acinar adenocarcinoma, suggesting that c-MET may not be a viable target for ADC drug development in prostatic acinar adenocarcinoma.

Objective:To investigate the expression of cellular repressor of E1A-stimulated gene 2 (CREG2) in colon adenocarcinoma (COAD) and its relationship with prognosis and tumor immune microenvironment.Methods:CREG2 mRNA expression data and survival data of patients were collected from 23 pan-cancer cohorts including COAD in the University of California, Santa Cruz (UCSC) database. The difference of CREG2 mRNA expression levels in cancer tissues and paracancerous tissues was compared. Univariate Cox proportional risk model was used to analyze the correlation of high and low expressions of CREG2 mRNA in pan-cancer tissues with the overall survival (OS) of patients; taking the pathological results as the gold standard, the optimal cut-off value of CREG2 mRNA expression level in cancerous and paracancerous tissues obtained by receiver operating characteristic (ROC) curve was used to group the high and low expressions. Kaplan-Meier method was used to draw the OS curve of pan-cancer CREG2 mRNA high and low expression groups, and log-rank test was used for comparison between groups. The data of patients with complete clinical data of the COAD cohort were further extracted from the UCSC database. The distribution of patients with high and low CREG2 mRNA expressions stratified by different clinicopathological characteristics of different groups was compared. Kaplan-Meier method was used to analyze the OS of patients with high and low CREG2 mRNA expressions as well as the OS of patients stratified by different clinicopathological characteristics, and log-rank test was used for comparison between groups; multivariate Cox proportional risk model was used to screen the independent factors influencing the OS of COAD patients. The chips of cancer tissues and the corresponding paracancerous tissues from 58 COAD patients were collected for immunohistochemical staining. CREG2 protein expression level between the cancer tissues and paracancerous tissues was compared. The product of the proportion of positive cells and the staining intensity score was treated as the CREG2 protein expression level in the tissues; taking pathological results as the gold standard, the ROC curve was used to analyze the therapeutic efficacy of CREG2 protein level in judging cancer tissues and paracancerous tissues. Based on the tumor immune assessment resource (TIMER) database, Pearson function was used to analyze the correlation between the CREG2 mRNA expression and the degree of immune cell infiltration in COAD tissues. ESTIMATE algorithm was used to obtain each immune infiltration score, and the immune infiltration score was compared between the CREG2 mRNA high and low expression groups.Results:In the 23 pan-cancer cohorts of UCSC database, the CREG2 mRNA expression level in 17 cohort cancer tissues including COAD was higher than that in paracancerous tissues, and the CREG2 mRNA expression level in 3 cohort cancer tissues including glioma was lower than that in paracancerous tissues, and the differences were statistically significant (all P < 0.05). Univariate Cox regression analysis showed that the OS of patients with high CREG2 mRNA expression was worse than that in those with low CREG2 mRNA expression in 5 cohort cancer tissues including COAD (236 cases, HR = 1.13, 95% CI: 1.01-1.26, P = 0.030), and in glioma cohorts (553 cases, HR = 0.90, 95% CI: 0.85-0.95, P < 0.001), the OS of patients with low CREG2 mRNA expression was worse than that in those with high CREG2 mRNA expression. Kaplan-Meier analysis also showed that the OS of COAD, lung adenocarcinoma, renal papillary cell carcinoma, and hepatocellular carcinoma patients with high CREG2 mRNA expression was worse than that of those with low CREG2 mRNA expression, and the OS of glioma patients with high CREG2 mRNA expression was better than that of those with low CREG2 mRNA expression. In the UCSC database, CREG2 mRNA expression level in 222 COAD patients with complete clinical data was higher than that in paracancerous normal tissues. The proportion of patients with high CREG2 mRNA expression in mismatch repair deficient patients with COAD was higher than that in mismatch repair normal patients [87.2% (34/39) vs. 69.4% (127/183)], and the difference was statistically significant ( P = 0.024). There were no statistically significant differences in the proportion of patients with high CREG2 mRNA expression among patients stratified by age, gender and pathological stage (all P > 0.05). Kaplan-Meier analysis showed that patients with pathological stage Ⅲ-Ⅳ, stage T 4, lymph node metastasis, distant metastasis, and high CREG2 mRNA expression had poor OS, and the differences were statistically significant (all P < 0.05); and multivariate Cox regression analysis showed that distant metastasis ( HR = 3.61, 95% CI: 1.81-7.23, P < 0.01) and high CREG2 mRNA expression ( HR = 4.29, 95% CI: 1.69-10.93, P < 0.01) were independent risk factors for poor OS in COAD patients. The result of immunohistochemistry method detection of clinical samples showed that in cancer and paracancerous tissues, CREG2 protein was mainly expressed in the cytoplasm, and the expression level of CREG2 protein in COAD tissues was higher than that in corresponding paracancerous tissues (164±23 vs. 146±24, P < 0.001). ROC curve analysis showed that CREG2 protein expression in COAD tissues could be used to judge cancer and paracancerous tissues (the area under the curve = 0.720, 95% CI: 0.628-0.814, P < 0.001). The analysis of the data in TIMER database showed that CREG2 expression level in COAD tissues was correlated with infiltration levels of CD8 + T cells ( R = 0.158, P = 0.001), CD4 + T cells ( R = 0.125, P = 0.012), macrophages ( R = 0.202, P < 0.001), neutrophils ( R = 0.248, P < 0.001), and dentritic cells ( R = 0.265, P < 0.001). The matrix score and estimated score in the high CREG2 expression group were higher than those in the low expression group, and the differences were statistically significant (all P < 0.05). Conclusions:CREG2 gene is highly expressed in COAD tissues, and its high expression may be related to the poor prognosis of COAD patients. The microenvironment of COAD with high CREG2 expression has a higher abundance of stromal cells, and CREG2 may be involved in immune regulation and immune response.

Objective To report the diagnosis and treatment processes of a fungal keratitis case caused by Lasio-diplodia theobromae(L.theobromae),and enhance the diagnosis and treatment experience on fungal keratitis caused by this rare pathogen.Methods Corneal scraping specimen from a patient with fungal keratitis was collec-ted.Gram-staining and fluorescence staining were conducted on specimen,followed with direct microscopic observa-tion and isolation culture.The strain was identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)technology and targeted DNA sequencing.Antimicrobial susceptibility testing was conducted.Literatures were summarized and clinical data on fungal keratitis caused by this pathogen were sorted out.Results Septal fungal hyphae could be seen on the corneal scraping specimen microscopically.The strain was identified as L.theobromae by both MALDI-TOF MS and targeted DNA sequencing after culture.Antimicrobial susceptibility testing(microdilution method)showed that the minimal inhibitory concentrations(MICs)of ampho-tericin B,vorionazole,itraconazole,posaconazole,fluconazole,5-fluorocytosine,micafungin,caspofungin,and anidulafungin against this strain were 1,0.25,＞8,0.25,＞256,8,16,2,and 0.25 μg/mL,respectively.Pa-tient recovered well after antifungal treatment plan was adjusted according to antimicrobial susceptibility testing re-sults.Conclusion L.theobromae is a rare pathogen that causes fungal keratitis.Laboratory tests provide rapid mi-croscopic examination results,and take MALDI-TOF MS and targeted DNA sequencing identification techniques as effective means to detect rare pathogen.In vitro antimicrobial susceptibility testing result can provide reference for clinicians to correctly use antifungal agents for treatment of infection due to this pathogen.

Objective : To investigate whole genome information of a newly isolatedBifidobacterium animalissubsp. lactic B4 strain from healthy human feces was analyzed and its probiotic properties. Methods : The antimicrobial resistance, hemolytic, gastric acid tolerance and biochemical characteristics of B. animalis B4 were evaluated byin vitroexperiments, and its whole genome was sequentially sequenced and functional annotation was performed by next and three-generation sequencing technology. Results: Whole genome sequencing of B. animalis B4 showed that its genome size was 1 944 146 bp, with GC content of 60.49%, no plasmid, and a total of 1 642 genes. The results ofin vitroanalysis showed that the B. animalis B4 had good probiotic properties, including non-hemolytic and stomach acid resistance. At the same time, the genome results showed that the B. animalis B4 strain did not have toxin and disease-related genes, drug resistance genes were few and the transmission ability was not high, so it had high safety. Gene annotation of KEGG, COG and GO showed that it contained many biological active enzymes, such as β-galactosidase, L-lactate dehydrogenase and other probiotic genes. Conclusion The B. animalis B4 has good probiotic properties, showing excellent safety at the genetic level, with a probiotic gene sequence.

Premature ovarian insufficiency(POI)is a complex endocrine disorder that significantly affects the physiological and repro-ductive functions of women,and it has become one of the main causes of infertility in women of childbearing age.The clinical features of POI include amenorrhea or oligomenorrhea,low estrogen or estrogen deficiency,and an increase in the level of gonadotropin.The in-cidence rate of POI tends to increase,yet its etiology and pathogenesis remain unclear.At present,the conventional treatment methods for POI have limited efficacy in fundamentally improving ovarian function or addressing the fertility issue.With the development of re-generative medicine,mesenchymal stem cells(MSCs)have become one of the research hotspots in POI therapy,and their exosomes have attracted wide attention as a promising"cell-free therapy".A number of animal experimental studies have shown that MSCs and their exosomes can exert a therapeutic effect on POI by affecting granulosa cell proliferation and apoptosis,promoting ovarian angiogen-esis,reducing oxidative stress and fibrosis,enhancing follicular development,and regulating immunity.In addition,related clinical studies have also made some progress.This article reviews the mechanisms and clinical effect of MSCs and their exosomes in the treatment of POI,in order to provide a reference for further research and bench-to-bedside translation of POI therapies.

This study explores the effect of total secondary ginsenosides(TSG) on apoptosis and energy metabolism in H9c2 cells under hypoxia and its potential mechanisms. H9c2 cell viability was observed and the apoptosis rate was calculated to determine suitable intervention concentrations of TSG, antimycin A complex(AMA), and coenzyme Q10(CoQ10), along with the duration of hypoxia. H9c2 cells at the logarithmic phase were divided into a normal group, a model group, a TSG group, an AMA group, a TSG+AMA group, and a CoQ10 group. All groups, except the normal group, were treated with their respective intervention drugs and cultured under hypoxic conditions. Adenosine triphosphate(ATP) content and creatine kinase(CK) activity were measured using an ATP chemiluminescence assay kit and a CK colorimetric assay kit. Flow cytometry was used to assess apoptosis rates, and Western blot evaluated the expression levels of apoptosis-related proteins, including B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), cysteinyl aspartate-specific protease(caspase)-3, caspase-8, and caspase-9, as well as mitochondrial biogenesis-related proteins peroxisome proliferator-activated receptor-γ coactivator 1α(PGC-1α), estrogen-related receptor-α(ERRα), nuclear respiratory factor(NRF)-1, NRF-2, peroxisome proliferator activated receptor-α(PPARα), and Na~+-K~+-ATPase. RT-PCR was employed to analyze the mRNA expression of mitochondrial biogenesis factors, including PGC-1α, ERRα, NRF-1, NRF-2, PPARα, mitochondrial transcription factor A(TFAM), mitochondrial cytochrome C oxidase 1(COX1), and mitochondrial NADH dehydrogenase subunit 1(ND1), ND2. The selected intervention concentrations were 7.5 μg·mL~(-1) for TSG, 10 μmol·L~(-1) for AMA, and 1×10~(-4) mol·L~(-1) for CoQ10, with a hypoxia duration of 6 h. Compared with the normal group, the model group showed decreased ATP content and CK activity, increased apoptosis rates, decreased Bcl-2 expression, and increased Bax, caspase-3, caspase-8, and caspase-9 expression in H9c2 cells. Additionally, the protein and mRNA expression levels of mitochondrial biogenesis-related factors(PGC-1α, ERRα, NRF-1, NRF-2, PPARα), mRNA expression of TFAM, COX1, and ND1, ND2, and protein expression of Na~+-K~+-ATPase in mitochondrial DNA, were also reduced. In the TSG and CoQ10 groups, ATP content and CK activity increased, and apoptosis rates decreased compared with those in the model group. The TSG group showed decreased protein expression of apoptosis-related proteins Bax, caspase-3, caspase-8, and caspase-9, increased protein and mRNA expression of mitochondrial biogenesis factors PGC-1α, ERRα, NRF-1, and PPARα, and increased NRF-2 protein expression and TFAM mRNA expression in mitochondrial DNA. Conversely, in the AMA group, ATP content and CK activity decreased, the apoptosis rate increased, Bcl-2 expression decreased, and Bax, caspase-3, caspase-8, and caspase-9 expression increased, alongside reductions in PGC-1α, ERRα, NRF-1, NRF-2, PPARα protein and mRNA expression, as well as TFAM, COX1, ND1, ND2 mRNA expression and Na~+-K~+-ATPase protein expression. Compared with the TSG group, the TSG+AMA group exhibited decreased ATP content and CK activity, increased apoptosis rates, decreased Bcl-2 expression, and increased Bax, caspase-3, caspase-8, and caspase-9 expression, along with decreased PGC-1α, ERRα, NRF-1, NRF-2, and PPARα protein and mRNA expression and TFAM, COX1, and ND1, ND2 mRNA expression. Compared with the AMA group, the TSG+AMA group showed increased CK activity, decreased apoptosis rate, increased Bcl-2 expression, and decreased Bax, caspase-8, and caspase-9 expression. Additionally, the protein and mRNA expression of PGC-1α, ERRα, NRF-1, PPARα, mRNA expression of TFAM, COX1, ND1, ND2, and Na~+-K~+-ATPase protein expression increased. In conclusion, TSG enhance ATP content and CK activity and inhibit apoptosis in H9c2 cells under hypoxia, and the mechanisms may be related to the regulation of PGC-1α, ERRα, NRF-1, NRF-2, PPARα, and TFAM expression, thus promoting mitochondrial biogenesis.
Apoptosis/drug effects* ; Ginsenosides/pharmacology* ; Energy Metabolism/drug effects* ; Mitochondria/metabolism* ; Animals ; Rats ; Cell Line ; Cell Hypoxia/drug effects* ; Organelle Biogenesis ; Adenosine Triphosphate/metabolism* ; Humans ; Cell Survival/drug effects*

OBJECTIVE To determine the species,quantity,and dispersion patterns of airborne pollen in two districts and five counties of Handan City,Hebei Province.METHODS The gravity sedimentation method was employed from October 1,2023,to September 30,2024,for daily and consecutive pollen exposure slides in two districts(Congtai District and Yongnian District)and five counties(Qiu County,Daming County,Cheng'an County,Cixian County,and She County)of Handan City,Hebei Province.The types of pollen were counted and identified under an optical microscope,and the pollen data were statistically analyzed.RESULTS The annual pollen quantity in the two districts and five counties of Handan City ranged from 24 031 grains/1 000 mm2 to 85 131 grains/1 000 mm2.The dissemination of major allergenic pollen presented two peaks.The first peak occurred in spring(March to April),mainly composed of Platanus(25.15％),Broussonetia(13.61％),Pinus(8.48％);the second peak occurred in(August to September),mainly consisting of Humulus/Cannabis(13.71％)and Artemisia(8.44％).Pollen in spring was significantly higher than that in summer and autumn.CONCLUSION The major allergenic airborne pollen in the two districts and five counties of Handan City exhibited two peaks in spring and summer-autumn,with Platanus pollen,Broussonetia pollen,Pinus pollen,Humulus/Cannabis pollen,and Artemisia pollne being the dominant pollen types.Among them,Platanus pollen was the absolute dominant pollen in this region.