ObjectiveTo investigate the mechanism of action of Bushen Huoxue prescription （BSHXP） in regulating premature ovarian failure in rats through the PTEN-induced kinase 1 （PINK1）/Parkinson's protein （Parkin） signaling pathway-mediated mitophagy. MethodsA total of 48 rats were randomly divided into a blank group consisting of eight rats， while the remaining 40 rats underwent modeling. The modeling group was intraperitoneally injected with 4 mg·kg^-1 cisplatin solution， followed by a second injection one week later， for a total of two injections. The estrous cycle was observed through vaginal smears for 14 consecutive days to determine whether the modeling was successful. The successfully modeled rats were randomly divided into a model group， groups receiving low， medium， and high doses of BSHXP at 9.72， 19.44， and 38.88 g·kg^-1·d^-1 （BSHXP-L， BSHXP-M， and BSHXP-H groups）， and a positive control group treated with estradiol valerate （0.09 mg·kg^-1·d^-1）， for 21 consecutive days. The body weight of the rats was measured weekly. After the final administration， rats were anesthetized， and their blood and ovaries were collected. The ovarian weight was measured. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of anti-Müllerian hormone （AMH）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E₂）. Assay kits were used to measure the levels of superoxide dismutase （SOD） and malondialdehyde （MDA） in the rat serum. Hematoxylin-eosin （HE） staining was used to observe the morphological changes in the ovaries. Immunohistochemistry （IHC） was performed to detect microtubule autophagy-related protein 1 light chain 3B（LC3B） protein expression in ovarian tissue， and electron microscopy was employed to examine the mitochondrial and autophagosome changes in the rat ovaries. Western blot was used to detect the protein expression of PINK1， Parkin， LC3B， and p62. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of PINK1， Parkin， LC3B， and p62 in ovarian tissue. ResultsCompared with the blank group， the model group showed significant reductions in body weight， weight gain， and ovarian weight （P<0.01）， along with decreased serum AMH and E₂ levels （P<0.01）， while FSH and LH levels were increased （P<0.01）. Serum MDA levels were significantly increased （P<0.01）， and SOD levels were significantly reduced （P<0.01）. The ovarian tissue structure was disordered， and the zona pellucida was wrinkled into an irregular acidophilic annular object， accompanied by an increased number of closed follicles. Electron microscopy showed mitochondrial swelling， unclear structure， and no obvious autophagosomes and autolysosome structures. The proteins and mRNA expression levels of PINK1， Parkin， LC3B， and p62 in the ovarian tissue were significantly reduced （P<0.01）. Compared with the model group， all treatment groups showed varying degrees of increases in body weight and ovarian weight （P<0.05， P<0.01）. Except for the BSHXP-L group， all treatment groups showed increased body weight gain （P<0.01）. All treatment groups showed significantly increased serum AMH and decreased FSH levels （P<0.01）. Except for the BSHXP group， all treatment groups showed varying degrees of increase and decrease in serum E₂ and LH levels （P<0.05， P<0.01）. All treatment groups showed reduced serum MDA levels （P<0.01）， while the BSHXP-M， BSHXP-H， and the positive control groups demonstrated improved serum SOD levels （P<0.05， P<0.01）. All treatment groups showed an increased number of follicles at all stages， visible mature follicles， and a decreased number of closed follicles. Electron microscopy showed relieved mitochondrial swelling， morphology close to normal， clear structure， and visible formation of autolysosomes in all treatment groups. Additionally， the protein and mRNA expression levels of PINK1， Parkin， LC3B， and p62 in ovarian tissue were significantly increased （P<0.05， P<0.01）. ConclusionBSHXP may improve ovarian function in rats with premature ovarian failure by regulating the PINK1/Parkin signaling pathway， activating mitochondrial autophagy， and reducing oxidative damage.

ObjectiveTo investigate the mechanism of action of Bushen Huoxue prescription （BSHXP） in regulating premature ovarian failure in rats through the PTEN-induced kinase 1 （PINK1）/Parkinson's protein （Parkin） signaling pathway-mediated mitophagy. MethodsA total of 48 rats were randomly divided into a blank group consisting of eight rats， while the remaining 40 rats underwent modeling. The modeling group was intraperitoneally injected with 4 mg·kg^-1 cisplatin solution， followed by a second injection one week later， for a total of two injections. The estrous cycle was observed through vaginal smears for 14 consecutive days to determine whether the modeling was successful. The successfully modeled rats were randomly divided into a model group， groups receiving low， medium， and high doses of BSHXP at 9.72， 19.44， and 38.88 g·kg^-1·d^-1 （BSHXP-L， BSHXP-M， and BSHXP-H groups）， and a positive control group treated with estradiol valerate （0.09 mg·kg^-1·d^-1）， for 21 consecutive days. The body weight of the rats was measured weekly. After the final administration， rats were anesthetized， and their blood and ovaries were collected. The ovarian weight was measured. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of anti-Müllerian hormone （AMH）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E₂）. Assay kits were used to measure the levels of superoxide dismutase （SOD） and malondialdehyde （MDA） in the rat serum. Hematoxylin-eosin （HE） staining was used to observe the morphological changes in the ovaries. Immunohistochemistry （IHC） was performed to detect microtubule autophagy-related protein 1 light chain 3B（LC3B） protein expression in ovarian tissue， and electron microscopy was employed to examine the mitochondrial and autophagosome changes in the rat ovaries. Western blot was used to detect the protein expression of PINK1， Parkin， LC3B， and p62. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of PINK1， Parkin， LC3B， and p62 in ovarian tissue. ResultsCompared with the blank group， the model group showed significant reductions in body weight， weight gain， and ovarian weight （P<0.01）， along with decreased serum AMH and E₂ levels （P<0.01）， while FSH and LH levels were increased （P<0.01）. Serum MDA levels were significantly increased （P<0.01）， and SOD levels were significantly reduced （P<0.01）. The ovarian tissue structure was disordered， and the zona pellucida was wrinkled into an irregular acidophilic annular object， accompanied by an increased number of closed follicles. Electron microscopy showed mitochondrial swelling， unclear structure， and no obvious autophagosomes and autolysosome structures. The proteins and mRNA expression levels of PINK1， Parkin， LC3B， and p62 in the ovarian tissue were significantly reduced （P<0.01）. Compared with the model group， all treatment groups showed varying degrees of increases in body weight and ovarian weight （P<0.05， P<0.01）. Except for the BSHXP-L group， all treatment groups showed increased body weight gain （P<0.01）. All treatment groups showed significantly increased serum AMH and decreased FSH levels （P<0.01）. Except for the BSHXP group， all treatment groups showed varying degrees of increase and decrease in serum E₂ and LH levels （P<0.05， P<0.01）. All treatment groups showed reduced serum MDA levels （P<0.01）， while the BSHXP-M， BSHXP-H， and the positive control groups demonstrated improved serum SOD levels （P<0.05， P<0.01）. All treatment groups showed an increased number of follicles at all stages， visible mature follicles， and a decreased number of closed follicles. Electron microscopy showed relieved mitochondrial swelling， morphology close to normal， clear structure， and visible formation of autolysosomes in all treatment groups. Additionally， the protein and mRNA expression levels of PINK1， Parkin， LC3B， and p62 in ovarian tissue were significantly increased （P<0.05， P<0.01）. ConclusionBSHXP may improve ovarian function in rats with premature ovarian failure by regulating the PINK1/Parkin signaling pathway， activating mitochondrial autophagy， and reducing oxidative damage.

OBJECTIVE: To evaluate the effect of palliative care on drug use, medical service utilization and medical expenditure of patients with advanced cancer. METHODS: A cohort of patients including pal-liative care and standard care was constructed using the medical records of the patients in Peking University Cancer Hospital from 2018 to 2020, and coarsened exact matching was used to match the two groups of patients. The average monthly opioid consumption, hospitalization rate, intensive care unit (ICU) rate and operation rate, and the average monthly total cost were selected to evaluate drug use, medical service utilization and medical expenditure. Chi-square test and Wilcoxon signed rank test were used to compare the differences between the two groups before and after exposure and the change in the palliative care group. The net impact of palliative care on the patients was calculated using the difference-in-differences analysis. RESULTS: In this study, 180 patients in the palliative care group and 3 101 patients in the stan-dard care group were finally included in the matching, and the matching effect of the two groups was good (L₁ < 0.1). Before and after exposure, the average monthly opioid consumption in the palliative care group was significantly higher than that in the standard care group (Before exposure: 0.3 DDD/person-month vs. 0.1 DDD/person-month, P < 0.01; After exposure: 0.7 DDD/person-month vs. 0.1 DDD/person-month, P < 0.01; DDD refers to defined daily dose), palliative care significantly increased the average monthly opioid consumption in the patients (0.3 DDD/person-month, P < 0.01). The hospitalization rate (48.9% vs. 74.3%, P < 0.01) and operation rate (3.9% vs. 8.8%, P < 0.01) of the patients in palliative care group were significantly lower than those in standard care group, and the ICU rate became similar between the two groups (1.1% vs. 1.6%, P=0.634). Palliative care significantly reduced the patients ' hospitalization rate (-25.6%, P < 0.01), ICU rate (-4.9%, P < 0.01) and operation rate (-14.5%, P < 0.01). Before and after exposure, the average monthly total costs of pal-liative care group were slightly higher than those of standard care group (Before exposure: 20 092.3 yuan vs. 19 132.8 yuan, P=0.725; After exposure: 9 719.8 yuan vs. 8 818.8 yuan, P=0.165). Palliative care increased the average monthly total cost by 2 208.8 yuan, but it was not statistically significant (P=0.316). CONCLUSION Palliative care can increase the opioid consumption in advanced cancer patients, reduce the rates of hospitalization, ICU and surgery, but has no significant effect on medical expenditure.
Humans ; Palliative Care/economics* ; Neoplasms/drug therapy* ; Analgesics, Opioid/economics* ; Male ; Female ; Middle Aged ; Aged ; Hospitalization/economics* ; Intensive Care Units/statistics & numerical data* ; Health Expenditures/statistics & numerical data* ; Adult ; Drug Utilization/statistics & numerical data* ; Patient Acceptance of Health Care/statistics & numerical data*

Natural products (NPs) have historically been a fundamental source for drug discovery. Yet the complex nature of NPs presents substantial challenges in pinpointing bioactive constituents, and corresponding targets. In the present study, an innovative natural product virtual screening-interaction-phenotype (NP-VIP) strategy that integrates virtual screening, chemical proteomics, and metabolomics to identify and validate the bioactive targets of NPs. This approach reduces false positive results and enhances the efficiency of target identification. Salvia miltiorrhiza (SM), a herb with recognized therapeutic potential against ischemic stroke (IS), was used to illustrate the workflow. Utilizing virtual screening, chemical proteomics, and metabolomics, potential therapeutic targets for SM in the IS treatment were identified, totaling 29, 100, and 78, respectively. Further analysis via the NP-VIP strategy highlighted five high-confidence targets, including poly [ADP-ribose] polymerase 1 (PARP1), signal transducer and activator of transcription 3 (STAT3), amyloid precursor protein (APP), glutamate-ammonia ligase (GLUL), and glutamate decarboxylase 67 (GAD67). These targets were subsequently validated and found to play critical roles in the neuroprotective effects of SM. The study not only underscores the importance of SM in treating IS but also sets a precedent for NP research, proposing a comprehensive approach that could be adapted for broader pharmacological explorations.

Objective To investigate the role of Kelch-like-epichlorohydrin-associated protein 1/nuclear factor erythroid-derived factor-2-related factor 2(Keap1-Nrf2)and nuclear factor-κB(NF-κB)signaling pathways in sepsis-associated encephalopathy(SAE).Methods Male C57BL/6J mice of SPF were randomly divided into four groups(n=10):the control group and LPS 6 h,24 h and 48 h groups.The behavioral changes of the mice were assessed based on their general conditions and open field test(OFT).ELISA was used to measure the levels of pro-inflammatory cytokines in mouse serum,and the antioxidant capacity assay kit to examine antioxidant activity in brain tissues of mice.Real-time quantitative PCR(qPCR)was adopted to detect the mRNA levels of toll-like receptor4(Tlr4),NF-κB,Keap1 and Nrf2 in the hippocampus,and to determine protein expressions of NF-κB a Nrf2、Keap1 and Tlr4 with Western blotting.Results Compared to the control group,the serum concentrations of interleukin 6(IL-6)in lipopolysaccharide(LPS)groups increased at 6 h,and reached the peak at 24 h and 48 h(P＜0.01).The levels of serum interleukin 18(IL-18)in the LPS groups increased significantly at 6 h and 24 h(P＜0.01)but there was no statistically significant difference compared with the 48h group.The results indicated the activity of superoxide dismutase(SOD)and glutathione(GSH)in brain tissues in LPS groups increased(P＜0.01).OFT results showed the time spent in the center of the open field,the distance covered around the center,and total distance covered by mice in LPS groups were significantly reduced(P＜0.01),except for the time spent in the center of the open field in the LPS 24 h group.The mRNA expressions of Tlr4 and(LPS 6 h,48 h)NF-κB in the hippocampus tissue of mice in LPS groups were elevated(P＜0.05),so were the mRNA expressions of Keap1 and Nrf2 in LPS 6 h group.Additionally,the protein expressions of NF-κB,Keap1 and Tlr4 increased in LPS groups,so did the protein expression of Nrf2 in LPS 24 h and 48 h groups(P＜0.05).Conclusion Keap1-Nrf2 and NF-κB signaling pathways may play a certain role in SAE.

Objective To determine the optimal conditions for CXCR4 upregulation by comparing the expression levels of chemokine(C-X-C motif)receptor 4(CXCR4)in MSCs cultured with varying concentrations of oxygen and hydrogen peroxide(H2O2).Methods MSCs were cultured with 0.1％,1％,or 3％O2 and 50 μmol/L H2O2 for different lengths of time(3,6,12,and 24 h).The mRNA and protein expressions of CXCR4 in MSCs were measured by real-time quantitative PCR(qPCR),Western blotting,and immunofluorescence staining.The viability and chemotactic ability of MSCs were measured using CCK-8,wound-healing and Transwell migration assays.Results Both hypoxia and H2O2 treatment were found to upregulate MSC expressions of CXCR4 to some extent.The mRNA and protein levels of CXCR4 were higher after 6-12 h of culture of MSCs with 3％O2,and significantly higher when treated with H2O2 for 6 h.Cell viability was significantly increased after culture with 3％O2 compared with the control group and both 3％O2 and H2O2 pretreatment could enhance chemotactic migration in MSCs.Conclusion Culture with 3％O2 and H2O2 pretreatment can upregulate CXCR4 expressions in MSCs and enhance migration in cells,with superior effects observed with 3％O2.Therefore,treatment with 3％O2 represents the best choice for upregulating the chemotactic ability of MSCs.

Objective To explore the activation of tumor necrosis factor-α(TNF-α)signaling pathway and the expressions of the associated inflammatory factors in NPHP1-defective renal tubular epithelial cells.Methods A human proximal renal tubular cell(HK2)model of lentivirus-mediated NPHP1 knockdown(NPHP1KD)was constructed,and the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors CXCL5,CCL20,IL-1β,IL-6 and MCP-1 were detected using RT-qPCR,Western blotting or enzyme-linked immunosorbent assay.A small interfering RNA(siRNA)was transfected in wild-type and NPHP1KDHK2 cells,and the changes in the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors were examined.Results NPHP1KDHK2 cells showed significantly increased mRNA expressions of TNF-α,C/EBPβ,CXCL5,IL-1β,and IL-6(P<0.05),protein expressions of phospho-p38 and C/EBPβ(P<0.05),and IL-6 level in the culture supernatant(P<0.05),and these changes were significantly blocked by transfection of cells with siRNA-C/EBPβ(P<0.05).Conclusion TNF-α signaling pathway is activated and its associated inflammatory factors are upregulated in NPHP1KDHK2 cells,and C/EBPβ may serve as a key transcription factor to mediate these changes.

An efficient vacuum suction system is a necessary prerequisite for the smooth operation of the oral diagnosis and treatment.During the use of the dental units,there is often a situation of vacuum suction weakness,resulting in the inability to discharge the mixture of blood,saliva,dental tissue and other mixtures in time,which affects the doctor's treatment field and increases the risk of aspiration pneumonia and cross-infection in patients.The working principle,pipeline system,filters and other aspects of the vacuum suction system that may affect the suction efficiency was analyzed.The causes and solutions of vacuum suction weakness were discussed,and operation suggestions were proposed to ensure the safe and effective use of equipment and ensure the safety of diagnosis and treatment.

Objective To explore the activation of tumor necrosis factor-α(TNF-α)signaling pathway and the expressions of the associated inflammatory factors in NPHP1-defective renal tubular epithelial cells.Methods A human proximal renal tubular cell(HK2)model of lentivirus-mediated NPHP1 knockdown(NPHP1KD)was constructed,and the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors CXCL5,CCL20,IL-1β,IL-6 and MCP-1 were detected using RT-qPCR,Western blotting or enzyme-linked immunosorbent assay.A small interfering RNA(siRNA)was transfected in wild-type and NPHP1KDHK2 cells,and the changes in the expressions of TNF-α,p38,and C/EBPβ and the inflammatory factors were examined.Results NPHP1KDHK2 cells showed significantly increased mRNA expressions of TNF-α,C/EBPβ,CXCL5,IL-1β,and IL-6(P<0.05),protein expressions of phospho-p38 and C/EBPβ(P<0.05),and IL-6 level in the culture supernatant(P<0.05),and these changes were significantly blocked by transfection of cells with siRNA-C/EBPβ(P<0.05).Conclusion TNF-α signaling pathway is activated and its associated inflammatory factors are upregulated in NPHP1KDHK2 cells,and C/EBPβ may serve as a key transcription factor to mediate these changes.