OBJECTIVES: To assess the efficacy and safety of Tiaozhou Ziyin (TZZY) recipe for treatment of premature ovarian insufficiency (POI) and explore the possible mechanisms. METHODS: We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets, which were verified by Western blotting. We tested the efficacy and safety of the recipe in 60 POI patients, who were randomized into control group (n=30) with Femoston treatment and TZZY group (n=30) with additional TZZY recipe treatment for 3 menstrual cycles. RESULTS: The core active ingredients of TZZY recipe included kaempferol, β-sitosterol, luteolin, and quercetin. The core targets included SRC, TP53, STAT3, PIK3CA, and MAPK3, which were involved in positive regulation of cell movement and protein phosphorylation, the cancer pathways and the PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had good binding ability with the core targets. In female rat models of POI, TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins. In the clinical trial, treatment with Femoston and Femoston plus TZZY recipe both significantly increased E₂ levels and reduced FSH and LH levels and Kupperman scores of the patients, and the combined treatment produced significantly stronger effects. Both treatments increased the number of antral follicles of the patients, but the combined treatment also significantly increased the levels of AMH. CONCLUSIONS The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients, multiple therapeutic targets and multiple pathways, and activating the PI3K /Akt pathway is one of its main mechanisms of action, to improve ovarian reserve function, alleviate clinical symptoms, and enhance clinical efficacy in POI patients.
Female ; Primary Ovarian Insufficiency/drug therapy* ; Humans ; Drugs, Chinese Herbal/therapeutic use* ; Animals ; Rats ; Molecular Docking Simulation ; Signal Transduction ; Sitosterols/therapeutic use* ; Kaempferols/therapeutic use*

Objective To investigate the expression of Acyl-CoA synthetase long-chain family member 4(ACSL4)in liver cancer and its role in regulating ferroptosis and proliferation of liver cancer cells.Methods Clinical samples of liver cancer and adjacent normal liver tissues were examined for malondialdehyde(MDA)contents and for expressions of mRNA and protein expressions of ACSL4 and proliferating cell nuclear antigen(PCNA)using RT-qPCR and Western blotting.Human liver cancer Huh-7 cells were treated with Erastin(a ferroptosis inducer),Fer-1(a ferroptosis inhibitor),or both,and the changes in expression levels of MDA,ACSL4 and PCNA were detected,and the cell proliferation was assessed with plate cloning assay.Results MDA contents were lower and ACSL4 and PCNA expressions were higher significantly in liver cancer tissues than in adjacent liver tissues.In Huh-7 cells,Erastin treatment significantly inhibited mRNA and protein expressions of ACSL4 and PCNA,suppressed cell proliferation,and increased MDA contents.Fer-1 alone did not produce significant effect on cell viability but reversed the effect of Erastin on ACSL4 and PCNA expressions,cell proliferation and MDA contents.Conclusion ACSL4 level is significantly overexpressed in liver cancer.Erastin increases MDA contents and down-regulates ACSL4 expression,thereby promoting ferroptosis and inhibiting proliferation of liver cancer cells,and these effects can be reversed by Fer-1.

Objective:To investigate the differences in terms of blood cerebrospinal fluid barrier immune surveillance injury by lead acetate and nano-lead sulfide exposure in order to provide basis for the study of their mechanism of nerve injury caused by exposure to lead and nano lead.Methods:In June 2015, forty-five SPF SD male rats were randomly divided into control group, lead acetate group (20 mg/kg) and nano-lead sulfide group (20 mg/kg), with 15 rats in each group. The rats were intragastric five times a week, for nine weeks. The numbers of CD4 + T lymphocytes in blood and cerebrospinal fluid were detected by flow cytometry. The levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and cerebrospinal fluid were detected by ELISA. The expressions and distribution of intercellular cell adhesion molecule-1 (ICAM-1) and CD4 + T lymphocytes in choroid plexus were detected by laser confocal fluorescence immunoassay. The mRNA expression levels of IL-4, IFN-γ and ICAM-1 in the choroid plexus were detected by real-time PCR. Results:Compared with the control group, the proportion of CD4 + T lymphocytes in blood of rats in lead acetate group was increased, the proportions of CD4 + T lymphocytes in cerebrospinal fluid of rats in lead acetate group and nano-lead sulfide group were increased, the contents of IL-4 and IFN-γ in serum of rats in lead acetate group and nano-lead sulfide group were increased, the content of IL-4 in cerebrospinal fluid of rats in lead acetate group and the contents of IL-4 and IFN-γ in cerebrospinal fluid of rats in nano-lead sulfide group were increased, the differences were statistically significant ( P<0.05). The fluorescence intensity of ICAM-1 and CD4 + T lymphocytes in choriochoroid plexus of rats in lead acetate group and nano-lead sulfide group were stronger than those in control group, and the fluorescence intensity of CD4 + T lymphocytes of rats in nano-lead sulfide group was weaker than that in lead acetate group. Compared with the control group, the mRNA expression levels of ICAM-1, IL-4 and IFN-γ in choriochoroids plexus of rats in lead acetate group and nano-lead sulfide group were increased, and the mRNA expression levels of ICAM-1 and IL-4 in nano-lead sulfide group were higher than those in lead acetate group, while the mRNA expression level of IFN-γ in nano-lead sulfide group was lower than that in lead acetate group ( P<0.05) . Conclusion:Exposure to lead and nano-lead sulfide can cause the increase of CD4 + T lymphocytes, IL-4, IFN-γ and ICAM-1, which may be related to the damage to the immune surveillance of the blood cerebrospinal fluid barrier. And there is a difference in the injury caused by lead and nano-lead sulfide exposure.

Objective To investigate the expression of Acyl-CoA synthetase long-chain family member 4(ACSL4)in liver cancer and its role in regulating ferroptosis and proliferation of liver cancer cells.Methods Clinical samples of liver cancer and adjacent normal liver tissues were examined for malondialdehyde(MDA)contents and for expressions of mRNA and protein expressions of ACSL4 and proliferating cell nuclear antigen(PCNA)using RT-qPCR and Western blotting.Human liver cancer Huh-7 cells were treated with Erastin(a ferroptosis inducer),Fer-1(a ferroptosis inhibitor),or both,and the changes in expression levels of MDA,ACSL4 and PCNA were detected,and the cell proliferation was assessed with plate cloning assay.Results MDA contents were lower and ACSL4 and PCNA expressions were higher significantly in liver cancer tissues than in adjacent liver tissues.In Huh-7 cells,Erastin treatment significantly inhibited mRNA and protein expressions of ACSL4 and PCNA,suppressed cell proliferation,and increased MDA contents.Fer-1 alone did not produce significant effect on cell viability but reversed the effect of Erastin on ACSL4 and PCNA expressions,cell proliferation and MDA contents.Conclusion ACSL4 level is significantly overexpressed in liver cancer.Erastin increases MDA contents and down-regulates ACSL4 expression,thereby promoting ferroptosis and inhibiting proliferation of liver cancer cells,and these effects can be reversed by Fer-1.

Objective:To investigate the differences in terms of blood cerebrospinal fluid barrier immune surveillance injury by lead acetate and nano-lead sulfide exposure in order to provide basis for the study of their mechanism of nerve injury caused by exposure to lead and nano lead.Methods:In June 2015, forty-five SPF SD male rats were randomly divided into control group, lead acetate group (20 mg/kg) and nano-lead sulfide group (20 mg/kg), with 15 rats in each group. The rats were intragastric five times a week, for nine weeks. The numbers of CD4 + T lymphocytes in blood and cerebrospinal fluid were detected by flow cytometry. The levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and cerebrospinal fluid were detected by ELISA. The expressions and distribution of intercellular cell adhesion molecule-1 (ICAM-1) and CD4 + T lymphocytes in choroid plexus were detected by laser confocal fluorescence immunoassay. The mRNA expression levels of IL-4, IFN-γ and ICAM-1 in the choroid plexus were detected by real-time PCR. Results:Compared with the control group, the proportion of CD4 + T lymphocytes in blood of rats in lead acetate group was increased, the proportions of CD4 + T lymphocytes in cerebrospinal fluid of rats in lead acetate group and nano-lead sulfide group were increased, the contents of IL-4 and IFN-γ in serum of rats in lead acetate group and nano-lead sulfide group were increased, the content of IL-4 in cerebrospinal fluid of rats in lead acetate group and the contents of IL-4 and IFN-γ in cerebrospinal fluid of rats in nano-lead sulfide group were increased, the differences were statistically significant ( P<0.05). The fluorescence intensity of ICAM-1 and CD4 + T lymphocytes in choriochoroid plexus of rats in lead acetate group and nano-lead sulfide group were stronger than those in control group, and the fluorescence intensity of CD4 + T lymphocytes of rats in nano-lead sulfide group was weaker than that in lead acetate group. Compared with the control group, the mRNA expression levels of ICAM-1, IL-4 and IFN-γ in choriochoroids plexus of rats in lead acetate group and nano-lead sulfide group were increased, and the mRNA expression levels of ICAM-1 and IL-4 in nano-lead sulfide group were higher than those in lead acetate group, while the mRNA expression level of IFN-γ in nano-lead sulfide group was lower than that in lead acetate group ( P<0.05) . Conclusion:Exposure to lead and nano-lead sulfide can cause the increase of CD4 + T lymphocytes, IL-4, IFN-γ and ICAM-1, which may be related to the damage to the immune surveillance of the blood cerebrospinal fluid barrier. And there is a difference in the injury caused by lead and nano-lead sulfide exposure.

0bjective To explore the effectiveness of caregivers based on health education under the guidance of timing theory in the nursing of elderly patients after hip replacement.Methods By convenient sampling method,eighty elderly patients undergoing hip replacement were divided into control group and observation group,40 cases in each group.The control group was given routine orthopedic care,and the observation group was based on health management under the guidance of the timing theory(total hip replacement,TIR)based on routine orthopedic care.The Harris Hip Score,Activities of Daily Living Scale(Barthel index),Caregiver Burden Inventory and Family Caregiver Task Inventory were used to compare and evaluate the application effect of health education of two groups of caregivers in the nursing of elderly patients after hip replacement.Results The patients were observed one month and three months after discharge,the Harris score and Barthel index of the observation group were significantly higher than those of the control group(P<0.01).At discharge,1 month after discharge and 3 months after discharge,the FCTI score(P<0.01)and CBI score(P<0.01)of family caregivers in the observation group were significantly lower than those in the control group.Conclusion Caregivers based on health education under the guidance of timing theory can effectively reduce the care burden of caregivers,enhance the care ability of caregivers,improve the rehabilitation effect of patients,and ameliorative the quality of life of patients.

Objective To explore the diagnostic value of Likert score and EPE grade score based on multiparameter magnetic resonance imaging(mpMRI)for extracapsular extension in prostate cancer(PCa).Methods The MR imaging and histopathology data from 272 PCa patients were analyzed retrospectively.All patients underwent mpMRI examination within 2 months before radical prostatectomy.Two radiologists with over 10 years of experience assessed the mpMRI images according to the Likert score and EPE grade score,respectively,and compared with pathological findings.The consistency between the two radiologists was evaluated by weighted Kappa test.The statistical analysis was performed using MedCalc 20.0 software.The sensitivity,specificity and other indicators were calculated to analyze the optimal cut-off value of Likert score and EPE grade score for diagnosing extracapsular extension in PCa.The area under the curve(AUC)was used to compare the diagnostic performance of the two scoring systems for extracapsular extension in PCa.Results Among 272 PCa patients,there were 45 cases with extracapsular extension and 227 cases without extracapsular extension.The weighted Kappa coefficients were 0.730 and 0.820 for Likert score and EPE grade score,respectively,indicating good consistency.The optimal cut-off values for diagnosing extracapsular extension in PCa were Likert score 3 and EPE grade score 2.The sensitivity and specificity were 68.8%and 77.5%for Likert score 3,and 64.4%and 84.5%for EPE grade score 2,respectively.Both Likert score(AUC=0.780)and EPE grade score(AUC=0.797)had high accuracy in predicting extracapsular extension in PCa,with no significant difference(P>0.05).Conclusion Both Likert score and EPE grade score have good diagnostic performance in detecting extracapsular extension in PCa,which provides important diagnostic basis for clinical staging of PCa.

The pathogenesis theory of"spleen deficiency and stasis toxin"in gastric cancer holds that spleen is the source of generation and transformation of qi and blood,that spleen deficiency is the internal basis of disease and throughout the disease.Stasis toxin is based on spleen deficiency,which is the fundamental pathogenesis of gastric cancer.In the pathological process of gastric cancer,a variety of metabolic substances in tumor cells and tumor microenvironment,mainly glucose metabolic reprogramming,undergo metabolic changes to reconstruct the phenotype and function of tumor-related macrophages,which is consistent with the pathogenesis theory of"spleen deficiency and stasis toxin".Therefore,this article focused on the reprogramming of glucose metabolism in tumor microenvironment to drive the phenotypic remodeling of tumor-related macrophages,explored the scientific connotation of the pathogenesis theory of"spleen deficiency and stasis toxin"of gastric cancer,and provided references for the theoretical and clinical research on the treatment of gastric cancer by TCM.

OBJECTIVE To investigate the neuroprotective effect of curculigoside （CUR） on rats with spinal cord injury （SCI） based on phosphatase and tensin homologue deleted on chromosome ten gene-induced putative kinase 1 （PINK1）/E3 ubiquitin ligase Parkin signaling pathway. METHODS Taking male SD rats as subjects， 15 rats were randomly selected as sham operation group； the rest rats were chosen to establish SCI model by spinal cord impact method， and then were divided into model group， CUR low-dose group （36 mg/kg CUR， gavage）， CUR high-dose group （72 mg/kg CUR， gavage） and CUR high-dose+3- methyladenine （3-MA） group （72 mg/kg CUR， gavage+20 mg/kg autophagy inhibitor 3-MA， intraperitoneal injection）， with 15 rats in each group. Rats in each group were given corresponding liquid/normal saline， once a day， for 28 consecutive days. Basso- Beattie-Bresnahan （BBB） score and Rivlin inclined plate experiment were performed on the 14th and 28th day after administration； the pathological changes of spinal cord tissue in rats were observed in each group； the apoptosis of spinal cord tissue， the levels of oxidative stress factors [malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione （GSH）]， and the protein expressions of brain-derived neurotrophic factor （BDNF）， glial fibrillary acidic protein （GFAP）， PINK1， Parkin， p62 and microtubule- associated protein light chain 3 （LC3） were all determined. RESULTS Compared with the sham operation group， obvious edema and bleeding in the spinal cord tissue of rats were observed in the model group， accompanied by a large number of inflammatory cell infiltration； BBB score and inclined plate angle， SOD and GSH levels， the protein expressions of BDNF， PINK1 and Parkin， and LC3Ⅱ/Ⅰ ratio were significantly reduced； the apoptosis rate， MDA level， the protein expressions of GFAP and p62 in spinal cord tissue were significantly increased （P＜0.05）. Compared with the model group， the edema， bleeding and infiltration of inflammatory cells in the spinal cord tissue of rats were reduced in the administration groups， and the above quantitative indicators had been significantly improved （P＜0.05）； 3-MA could significantly reverse the improvement effects of the above indexes by CUR （P＜0.05）. CONCLUSIONS CUR can promote the recovery of neurological and motor functions in SCI rats， improve the pathological injury of the spinal cord and inhibit apoptosis， which may be related to mitochondrial autophagy mediated by activating the PINK1/Parkin signaling pathway.

Objective To explore the application efficacy and safety of oblique ultrasound-guided techniques in PICC puncture,in order to provide guidance and references for clinical application.Methods Through convenient sampling,654 patients from a tertiary A hospital in Zhejiang Province from March to December 2022 were selected as the study subjects.The random numbers were generated through Excel table functions and they were randomly grouped into 3 groups:A,B,and C.The ultrasound short axis method,long axis method,and oblique axis method were employed to guide PICC puncture catheterization,respectively.The success rate of PICC puncture,the number of subcutaneous adjustments of the puncture needle,puncture time,and the occurrence of puncture complications(such as hematoma,puncture of the posterior wall of blood vessels,accidental injury to arteries,and accidental injury to nerves)were recorded during the catheterization process in 3 groups.Results A total of 654 patients completed the study,including 215 in group A,219 in group B,and 220 in group C.The success rate of first-time puncture in the group C(86.36％)was higher than that in group A(73.95％)and group B(63.93％),and there was a statistically significant difference among 3 groups(P＜0.001).The subcutaneous adjustment frequency of the puncture needle was 1(1,1)in group C,1(1,2)in group A,and 1(1,2)in group B.The difference between 3 groups was statistically significant(P＜0.001);the puncture time of group C was shorter than that of group A and group B,and the difference was statistically significant(P＜0.001).There was a statistically significant difference in the puncture time between 3 groups(P＜0.017);the pairwise comparison of the number of subcutaneous needle adjustments and the success rate of a puncture between 3 groups showed that there was a statistical difference between group C and group A,and between group C and group B(P＜0.017),while there was no statistical difference between group A and group B(P＞0.017).There was statistical significance(P＜0.05)among 3 groups in terms of complications such as accidental nerve injury and puncture of the contralateral vascular wall by puncture needle,but there was no statistical significance in terms of accidental arterial injury and hematoma occurrence among 3 groups.Conclusion Compared with the short axis approach and the long axis approach,the ultrasound oblique axis approach guided PICC puncture has statistical differences in the success rate of a puncture and the incidence of puncture complications,etc.It is recommended to use the ultrasound oblique axis approach during PICC puncture.