OBJECTIVE To explore the pharmacokinetic characteristics of 3 blood-absorbed components of Xiangshao sanjie oral liquid in rats with hyperplasia of mammary gland （HMG）. METHODS Female SD rats were divided into control group and HMG group according to body weight， with 6 rats in each group. The HMG group was given estrogen+progesterone to construct HMG model. After modeling， two groups were given 1.485 g/kg of Xiangshao sanjie oral liquid （calculated by crude drug） intragastrically， once a day， for 7 consecutive days. Blood samples were collected before the first administration （0 h）， and at 5， 15， 30 minutes and 1， 2， 4， 8， 12， 24 hours after the last administration， respectively. Using chlorzoxazone as the internal standard， the plasma concentrations of ferulic acid， paeoniflorin and rosmarinic acid in rats were detected by UPLC-Q/TOF-MS. The pharmacokinetic parameters [area under the drug time curve （AUC0－24 h， AUC0－∞）， mean residence time （MRT0－∞）， half-life （t1/2）， peak time （tmax）， peak concentration （cmax）] were calculated by the non-atrioventricular model using Phoenix WinNonlin 8.1 software. RESULTS Compared with the control group， the AUC0－24 h， AUC0－∞ and cmax of ferulic acid in the HMG group were significantly increased （P＜0.05）； the AUC0－24 h， AUC0－∞ ， MRT0－∞ ， t1/2 and cmax of paeoniflorin increased， but there was no significant difference between 2 groups （P＞0.05）； the AUC0－24 h and MRT0-∞ of rosmarinic acid were significantly increased or prolonged （P＜0.05）. C ONCLUSIONS In HMG model rats， the exposure of ferulic acid， paeoniflorin and rosmarinic acid in Xiangshao sanjie oral liquid all increase， and the retention time of rosmarinic acid is significantly prolonged.

OBJECTIVES: This study aims to investigate the genome-wide DNA methylation and transcriptome expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with systemic sclerosis (SSc) with interstitial lung disease (ILD), and to analyze the effects of DNA methylation on Wnt/β-catenin and chemokine signaling pathways. METHODS: PBMCs were collected from 19 patients with SSc (SSc group) and 18 healthy persons (control group). Among SSc patients, there were 10 patients with ILD (SSc with ILD subgroup) and 9 patients without ILD (SSc without ILD subgroup). The genome-wide DNA methylation and gene expression level were analyzed by using Illumina 450K methylation chip and Illumina HT-12 v4.0 gene expression profiling chip. The effect of DNA methylation on Wnt/β-catenin and chemokine signal pathways was investigated. RESULTS: Genome-wide DNA methylation analysis identified 71 hypermethylated CpG sites and 98 hypomethylated CpG sites in the SSc with ILD subgroup compared with the SSc without ILD subgroup. Transcriptome analysis distinguished 164 upregulated genes and 191 downregulated genes in the SSc with ILD subgroup as compared with the SSc without ILD subgroup. In PBMCs of the SSc group, 35 genes in Wnt/β-catenin signaling pathway were hypomethylated, while frizzled-1 (FZD1), mitogen-activated protein kinase 9 (MAPK9), mothers against DPP homolog 2 (SMAD2), transcription factor 7-like 2 (TCF7L2), and wingless-type MMTV integration site family, member 5B (WNT5B) mRNA expressions were upregulated as compared with the control group (all P<0.05). Compared with the SSc without ILD subgroup, the mRNA expressions of dickkopf homolog 2 (DKK2), FZD1, MAPK9 were upregulated in the SSc with ILD subgroup, but the differences were not statistically significant (all P>0.05). In PBMCs of the SSc group, 38 genes in chemokine signaling pathway were hypomethylated, while β-arrestin 1 (ARRB1), C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 16 (CXCL16), FGR, and neutrophil cytosolic factor 1C (NCF1C) mRNA expressions were upregulated as compared with the control group (all P<0.05). Compared with the SSc without ILD subgroup, the mRNA expressions of ARRB1, CXCL10, CXCL16 were upregulated in the SSc with ILD subgroup, but the differences were not statistically significant (all P>0.05). CONCLUSIONS There are differences in DNA methylation and transcriptome profiles between SSc with ILD and SSc without ILD. The expression levels of multiple genes in Wnt/β- catenin and chemokine signaling pathways are upregulated, which might be associatea with the pathogenesis of SSc.
Humans ; DNA Methylation ; Transcriptome ; beta Catenin ; Leukocytes, Mononuclear ; Ligands ; DNA ; RNA, Messenger/genetics*

Objective:To investigate the characteristics of wideband acoustic immittance（WAI） measurements in patients with unilateral Ménière's disease（MD） and evaluate the clinical value of WAI in diagnosis of MD. Methods:WAI was performed in 30 patients with unilateral MD（30 ears for symptomatic and 30 ears for asymptomatic） and in 26 healthy individuals（52 ears）（control group）. The WAI measurements, including the frequency first appearing two peaks in energy absorbance（EA） tympanogram, resonance frequency（RF）, the peak value of absorbance（PVA）, the integral area of absorbance（IAA）, EA curve at peak pressure, were analyzed. Results:①The occurrence of two peaks in EA tympanogram in both the MD symptomatic and asymptomatic ear was observed in 27 ears（84.4%）, and 38 ears（70.4%） in the control group, with no significant difference in the frequency of first appearing in two peaks onset between the groups（all P>0.05）. ②The RF of the MD symptomatic ears was significantly lower than that of the asymptomatic ears（t=-3.544, P=0.001） and that of the control subjects（t=2.084, P=0.041）; there was no difference of RF between the MD asymptomatic ears and the control group（P>0.05）. ③The PVA were significantly lower in both MD symptomatic（t=4.240, P<0.01） and asymptomatic ears（t=4.202, P=0.001） than in controls. ④The IAA in MD symptomatic（t=3.295, P=0.001） and asymptomatic ears（t=3.193, P=0.003） was significantly lower than in the control group. ⑤Comparison of the EA curve at peak pressure of the three groups: the EAs of MD symptomatic ears were lower than those of the control group at the range of 1 059-2 911 Hz（all P<0.05）; the EAs of MD symptomatic ears were lower than those of MD asymptomatic ears within 1 000 Hz and 1 834-2 119 Hz（all P<0.05）; the EAs of MD asymptomatic ears were lower than those of the control group at the range of 515-2 748 Hz（all P<0.05）. Conclusion:Symptomatic ears in unilateral MD patients show alterations in some WAI measurements compared to asymptomatic ears and/or controls, suggesting that middle ear mechanical fuction of the affected side may be modified due to the endolymphatic hydrops. The clinical significance of WAI needs to be further explored in the context of evaluating MD.
Humans ; Meniere Disease/diagnosis* ; Endolymphatic Hydrops/diagnosis* ; Ear ; Hearing Tests ; Acoustics

【Objective】 To explore the expressions of PBRM1 and PD-L1 in renal cell carcinoma (RCC), and the molecular mechanism of PBRM1 regulating PD-L1, in order to provide experimental results for clinical immunotherapy. 【Methods】 The protein expressions of PBRM1 and PD-L1 were detected with immunohistochemistry, and their mRNA expressions were determined by analyzing TCGA database. Meanwhile, the relationship between overall survival and mRNA expressions of PBRM1 and PD-L1 were analyzed in TCGA database. The exosomes were extracted with exoEasy Maxi Kit. Expressions of exosomal biomarkers CD63 and CD9 were detected with Western blotting, and their morphology was observed with transmission electron microscope. PD-L1 expression after IFN-γ, IL-6 and TNF-α treatment was detected with Western blotting. 【Results】 The expression of PBRM1 was significantly lower in canver tissue than in paracancerous tissue (P<0.001). The loss rate of PBRM1 was up to 76.4%. PBRM1 expression was not correlated with PD-L1 expression. PBRM1 deletion activated TNF-α/exosome signaling pathway, leading to increase of PD-L1 secretion in exosomes, which was then transported to the outside of cells. 【Conclusion】 There is no relationship between PBRM1 and PD-L1 protein expressions in RCC. However, PBRM1 mutation can lead to inflammatory signaling pathway activation, inducing PD-L1 secretion, which is encapsulated in exosomes and transported to the outside of cells, and affects the response of immunotherapy.

【Objective】 To evaluate the predictive value of isoform [-2] proprostate-specific antigen, p2 PSA (p2PSA) and its derived indexes for prostate cancer in a Chinese cohort with PSA 4-20 ng/mL. 【Methods】 A total of 139 males scheduled for biopsy were enrolled in the prospective study from Nov.2021 to Jun.2022. The total PSA (tPSA), free PSA (fPSA), fPSA/tPSA (f/t) and p2PSA were collected, and the percentage of p2PSA(%p2PSA) and prostate health index(PHI) were calculated. The predictive value of p2PSA and its derived indexes were compared with traditional indexes with receiver operating characteristic (ROC) curve and Logistic analysis. 【Results】 Prostate cancer was found in 54 cases (38.8%). There were significant statistical differences in tPSA(10.68 vs.8.14, P=0.021), f/t(0.13 vs.0.16, P=0.006), p2PSA(30.25 vs.19.81, P<0.001), %p2PSA(21.52 vs.13.15, P<0.001) and PHI(64.3vs.38.2, P<0.001) between prostate cancer patients and non-prostate cancer patients. The area under the ROC curve (AUC) of tPSA, fPSA, %fPSA, p2PSA, %p2PSA and PHI were 0.63, 0.51, 0.63, 0.71, 0.73, and 0.80, respectively. The inclusion of %p2PSA and PHI significantly increased the prediction efficiency of the basic prediction model (AUCbase+PHI=0.81, AUCbase+%p2PSA=0.78, AUCbase=0.67). With 35 as the recommended cut-off value of PHI, the incidence of meaningless puncture was reduced by 25.8%(36/139). 【Conclusion】 The application of p2PSA and its derived indexes have good predictive value for patients with PSA 4-20 ng/mL. The combined detection of %p2PSA and PHI can significantly increase the detection efficiency of prostate cancer and reduce the incidence of meaningless prostate puncture by 25.8%.

Objective To investigate the effect of compound Fufangteng mixture-containing serum on the proliferation of bone marrow mesenchymal stem cell (BMSC) and its mechanism. Methods Rat BMSC were isolated, cultured and purified in vitro by direct adherence method. Cell morphology was observed. Surface markers were identified by flow cytometry. The rats were treated with compound Fufangteng mixture at a dose of 3 mL/(kg·d) by gavage for 14 d, and then the drug-containing serum was collected. BMSC were divided into the blank control group, drug-containing serum group, Notch1 small interfering ribonucleic acid (siRNA) group and Notch1 siRNA+drug-containing serum group. The proliferation rate of BMSC was detected and the relative expression levels of Notch1 signaling pathway-associated messenger ribonucleic acid (mRNA) and proteins were measured in each group. Results Microscopic observation showed that the first generation BMSC were seen in the long spindle shape, and grown in the parallel or spiral pattern. The third generation BMSC positively expressed CD90 and CD44, whereas were negative for CD45. Compared with the blank control group, the proliferation rate of BMSC in the drug-containing serum group and Notch1 siRNA+ drug-containing serum group was significantly increased, whereas that of BMSC was significantly decreased in the Notch1 siRNA group (all P < 0.05). Compared with the Notch1 siRNA group, the proliferation rate of BMSC was significantly increased in the Notch1 siRNA+drug-containing serum group (P < 0.05). Compared with the blank control group, the relative expression levels of Hey1 and Delta-like ligand (DLL)1 mRNA and proteins were significantly up-regulated in the drug-containing serum group, whereas those were significantly down-regulated in the Notch1 siRNA group and Notch1 siRNA+drug-containing serum group (all P < 0.05). Compared with the Notch1 siRNA group, the relative expression levels of Hey1 and DLL1 mRNA and proteins were significantly up-regulated in the Notch1 siRNA+drug-containing serum group (all P < 0.05). Conclusions Compound Fufangteng mixture-containing serum may promote the proliferation of rat BMSC, and its mechanism is probably associated with the activation of Notch1 signaling pathway.

Objective: The etiology of schizophrenia is unknown and is associated with abnormal spontaneous brain activity. There are no consistent results regarding the change in spontaneous brain activity of people with schizophrenia. In this study, we determined the specific changes in the amplitude of low-frequency fluctuation/fractional amplitude of low-frequency fluctuation (ALFF/fALFF) and regional homogeneity (ReHo) in patients with drug-naïve first-episode schizophrenia (Dn-FES). Methods: A comprehensive search of databases such as PubMed, Web of Science, and Embase was conducted to find articles on resting-state functional magnetic resonance imaging using ALFF/fALFF and ReHo in schizophrenia patients compared to healthy controls (HCs) and then, anatomical/activation likelihood estimation was performed. Results: Eighteen eligible studies were included in this meta-analysis. Compared to the spontaneous brain activity of HCs, we found changes in spontaneous brain activity in Dn-FES based on these two methods, mainly including the frontal lobe, putamen, lateral globus pallidus, insula, cerebellum, and posterior cingulate cortex. Conclusion We found that widespread abnormalities of spontaneous brain activity occur in the early stages of the onset of schizophrenia and may provide a reference for the early intervention of schizophrenia.

OBJECTIVE：To study the effec ts of Tibetan Codonopsis tralictrifolia extract （called“ZDS”for short ） on collagen-induced arthritis （CIA）model rats and its mechanism. METHODS ：Eight of 48 rats were randomly selected as normal control group （normal saline ），and the remaining 40 rats were used to establish CIA model. After successful modeling ，the rats were randomly divided into model group （normal saline ），ZDS low-dose ，medium-dose and high-dose groups （0.44，0.88，1.76 g/kg，by crude drug ），dexamethasone group （positive control ，0.002 5 g/kg），with 8 rats in each group. They were given relevant medicine intragastrically ，the volume of 400 μL，once a day ，for consecutive 28 days. The body weight of rats were weighed before medication （0 d），7，14，21 and 28 days after medication ；and arthritis indexes were scored. The pathological changes of the knee joint synoviual tissue were observed after last medication. The thymus index ，spleen index ，the levels of serum inflammatory factors （IL-1β，TNF-α，IL-6），protein expressions of NF-κB p65，p-NF-κB p65，IκB and p-IκB in synovial tissue were detected. RESULTS ：Compared with normal control group ，the body weight （14，21，28 days after administration ）of rats in model group was significantly reduced （P＜0.05）；the arthritis index score （before administration and different administration time）was significantly increased （P＜0.05）；the joint synovial tissue was pathologically damaged ；the thymus and spleen index ， inflammation factor level ，the protein expression of p-NF-κB p65 and p-IκB were increased significantly（P＜0.05），while the protein expression of IκB was decreased significantly（P＜0.05）. Compared with model group ，the level of IL- 1β was decreased significantly in ZDS low-dose group （P＜0.05）. Body weight of rats （21，28 days after administration ）were increased significantly in ZDS medium-dose and high-dose groups ，dexamethasone group （P＜0.05），while arthritis index score （14，21，28 days after administration ） was decreased significantly （P＜0.05）. The pathological damage of joint synovial tissue was significantly relieved ；thymus and spleen index ，inflammation factor level ，the protein expression of p-NF-κB p65 and p-IκB were significantly reduced，while the protein expression of IκB was significantly increased （P＜0.05）. CONCLUSIONS ：ZDS can improve CIA model rats to some extent ，and its mechanism may be related to the inhibition of NF-κB signaling pathway.

Objective:To evaluate the role of silent information regulator 1 (SIRT1)/nuclear factor E2 related factor 2 (Nrf2) signaling pathway in sleep deprivation-induced cognitive dysfunction in rats.Methods:Thirty-six adult male Sprague-Dawley rats, aged 54-56 weeks, weighing 600-750 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), sleep deprivation group (group SD) and sleep deprivation+ SIRT1 agonist Srt1720 group (group SD+ Srt). Sleep deprivation model was established by modified multi-platform water environment method.Srt1720 10 mg/kg was injected intraperitoneally every 12 h starting from 24 h before establishing the model in group SD+ Srt, while the equal volume of 0.9% normal saline was injected intraperitoneally in C and SD groups.After the end of sleep deprivation, Morris water maze test was performed to evaluate the cognitive function.Animals were then sacrificed, and their hippocampi were removed for determination of neuronal degeneration rate in hippocampal CA1 region (using HE staining), the apoptosis rate in hippocampal CA1 (using TUNEL assay ), the expression of SIRT1 and Nrf2 in hippocampal CA1 (by immunohistochemistry) and the contents of reactive oxygen species (ROS) and superoxide dismutase (SOD) (by microplate method). Results:Compared with group C, the escape latency was significantly prolonged, the time of staying at the platform quadrant was shortened, and the frequency of crossing the platform was decreased on 2-4 days, the apoptotic rate and neuronal degeneration rate were increased, the expression of SIRT1 and Nrf2 was down-regulated, ROC content was increased, and SOD content was decreased in SD and SD+ Srt groups ( P<0.05). Compared with group SD, the escape latency was significantly shortened, the time of staying at the platform quadrant was prolonged, and the frequency of crossing the platform was increased on 3 and 4 days, the apoptotic rate and neuronal degeneration rate were decreased, the expression of SIRT1 and Nrf2 was up-regulated, ROC content was decreased, and SOD content was increased in group SD+ Srt ( P<0.05). Conclusions:Sleep deprivation can induce oxidative stress response in hippocampus by inhibiting the activation of SIRT1/Nrf2 signaling pathway, leading to cognitive dysfunction in rats.

Objective:To examine treatment outcomes of breast phyllodes tumors and the prognosis factors of local recurrence.Methods:This retrospective cohort study included 276 patients who underwent surgical resection at Breast Center, Peking University People′s Hospital from January 2011 to December 2019. Tumor subtype and histopathological features were determined from pathology reports, and the deadline of follow-up was September 30 th, 2020. All 276 patients underwent open surgery, including 17 patients of mastectomy, and 259 patients of lumpectomy. The enrolled patients were all female, with age of (41.5±11.3) years (rang: 11 to 76 years), and tumor diameter of 35(28) mm ( M( Q R)). The Kaplan-Meier method and Log-rank test were used for survival analysis. The multivariate analysis was implemented using the Cox proportional hazard model. Results:According the pathologic test, there were 191 patients of benign phyllodes tumor, 67 patients of borderline tumor and 18 patients of malignant tumor. There were 249 patients with a follow-up of more than 6 months, and 14.1% (35/249) had local recurrence. The time-to-recurrence was (28.6±22.2) months (range: 2 to 96 months), (29.1±18.1) months (range: 2 to 80 months), (32.1±30.1) months (range: 5 to 96 months) and (12.0±6.9) months (range: 8 to 20 months) for benign, borderline and malignant phyllodes tumors. Tumor diameter (≥100 mm vs.<50 mm, HR=3.968, 95%CI: 1.550 to 10.158, P=0.004) and malignant heterologous element (yes vs. no, HR=26.933, 95%CI: 3.105 to 233.600, P=0.003) were prognosis factors of local recurrence. One death from malignant phyllodes occurred after distant metastasis. The 3-year disease-free survival rates of benign, borderline and malignant phyllodes tumor were 88.2%, 81.7% and 81.4% ( P=0.300). Conclusion:Phyllodes tumors have a considerable local recurrence rate, which may be associated with tumor diameter and malignant heterologous element.