Charcoal-processed traditional Chinese herbal medicine has various therapeutic effects， including astringing， hemostasis， anti-diarrhea， clearing heat， tonifying， and warming the interior. This paper summarizes the clinical application features， compatible experiences， dosages， and precautions for over 20 types of charcoal-processed herbal medicine in the treatment of gynecological bleeding disorders caused by dysfunctions such as dysfunctional uterine bleeding， endometriosis， uterine incision pseudocavity， and vaginal bleeding resulting from threatened miscarriage. The charcoal-processed herbal medicine include Huangqin （Scutellaria Baicalensis） Charcoal， Dahuang （Rheum Palmatum） Charcoal， Cebai （Platycladus Orientalis） Charcoal， Diyu （Sanguisorba Officinalis） Charcoal， Daji （Cirsium Setosum） Charcoal， Xiaoji （Cirsium Japonicum） Charcoal， Shengdi （Rehmannia Glutinosa） Charcoal， Aiye （Artemisia Argyi） Charcoal， Paojiang （Zingiber Officinale） Charcoal， Xuduan （Dipsacus Asper） Charcoal， Duzhong （Eucommia Ulmoides） Charcoal， Qiancao （Rubia Cordifolia） Charcoal， Puhuang （Typha Angustifolia） Charcoal， Shanzha （Crataegus Pinnatifida） Charcoal， Jingjie （Schizonepeta Tenuifolia） Charcoal， Xueyu （Carthamus Tinctorius） Charcoal， Zonglyu （Areca Catechu） Charcoal， Wumei （Prunus Mume） Charcoal， Shudahuang （Rheum Officinale） Charcoal， Lianfang （Nymphaea Alba） Charcoal， Mianmaguanzhong （Clematis Armandii） Charcoal， and Oujie （Nelumbo Nucifera） Charcoal.

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.

This article summarized Professor You Zhaoling's clinical experience in the treatment of polycystic ovary syndrome(PCOS).It is believed that kidney deficiency and blood stasis are the basic etiology and pathogenesis,with liver and spleen disorders.Clinical treatment is based on tonifying the kidney and replenishing essence,promoting blood circulation and resolving stasis,supplemented by soothing the liver,strengthening the spleen,and resolving dampness;treatment should also be based on the patients'stage of illness,age,and body type;basic body temperature and B-ultrasound should be skillfully uses to monitor ovulation,and four methods of regulating bubbles were creatively created;an original hypothetical menstrual stimulation method for PCOS patients with menstrual disorders was created.Professor You's clinical experience has demonstrated the characteristics of TCM treatment,providing a new approach for the TCM treatment of PCOS.

Objective:To establish a nomogram prediction model for recurrence within 2 years after radical resection of hepatocellular carcinoma (HCC) based on clinical and ultrasonographic characteristics.Methods:Clinical data from 405 HCC patients (including 327 males and 78 females), aged 60 (53, 66) years old, who underwent radical hepatectomy in the Zhongshan Hospital, Fudan University, from January to December 2021, were retrospectively collected. The patients were divided into two groups: the training group ( n=283) and the validation group ( n=122). Based on recurrence within 2 years after surgery, the 283 patients in the training group were further categorized into the recurrence group ( n=73) and the non-recurrence group ( n=210). Among the 122 patients in the validation group, 33 had recurrence within 2 years, while 89 did not. Data on age, microvascular invasion, alpha-fetoprotein (AFP), AFP lentil lectin-reactive fraction (AFP-L3), protein induced by vitamin K absence or antagonist-II (PIVKA-II), tumor number, and enhancement homogeneity were collected. Logistic regression analysis was performed on the training group to identify risk factors associated with postoperative recurrence, and a nomogram model for predicting HCC recurrence was constructed based on these factors. Calibration curves were used to compare the consistency between predicted and actual outcomes in both the training and validation groups. Results:Multivariate logistic regression analysis revealed that younger age ( OR=0.976, 95% CI: 0.953-1.000, P=0.004), higher AFP-L3 ( OR=1.066, 95% CI: 1.014-1.120, P=0.012), higher PIVKA-II ( OR=1.000, 95% CI: 1.000-1.001, P=0.042), multiple tumors ( OR=0.399, 95% CI: 0.225-0.706, P=0.038), and heterogeneous enhancement ( OR=0.472, 95% CI: 0.243-0.916, P=0.045) were significant risk factors for recurrence after partial hepatectomy in HCC patients. The nomogram constructed based on these variables had a C-index of 0.87 (95% CI: 0.81-0.93) in the training group and 0.83 (95% CI: 0.77-0.89) in the validation group. The calibration curves for predicting recurrence within 2 years after partial hepatectomy in HCC patients showed a high degree of fit in both the training and validation groups, indicating a good agreement between predicted and actual outcomes. Conclusion:The nomogram model constructed based on preoperative clinical and ultrasonographic characteristics can effectively predict the risk of recurrence within 2 years after radical resection of HCC.

Objective:To analyze the viscosity characteristics of liver tumors and investigate the clinical value of shear wave dispersion (SWD) in the differentiation of benign and malignant liver tumors.Methods:A total of 103 patients with focal liver lesions were prospectively collected in Zhongshan Hospital Affiliated to Fudan University from October 2020 to July 2021, including 80 cases with single lesion and 23 cases with multiple lesions, and only the largest lesion was observed in patients with multiple lesions. SWD values were measured within the tumor and in the liver parenchyma 2 cm away from the tumor, and were compared between benign tumor group and maligant tumor group. The ROC curves of SWD value, SWD ratio and their combination in differentiating benign and malignant liver tumors were plotted respectively, and the optimal diagnostic threshold, the sensitivity, specificity and accuracy of different diagnostic methods were analyzed.Results:Among the 103 patients, 35 were benign and 68 were malignant. The SWD value of liver benign tumor group was lower than that of liver malignant tumor group [(16.38±3.58)m·s -1·kHz -1 vs (18.59±3.12)m·s -1·kHz -1], the SWD value of liver parenchyma background in liver benign tumor group was lower than that in liver malignant tumor group [(10.88±3.37)m·s -1·kHz -1 vs (14.31±3.34)m·s -1·kHz -1], and the differences were statistically significant (all P<0.05). The SWD ratio of benign tumor to surrounding liver parenchyma was higher than that of malignant tumor group [1.57(1.25, 2.00) vs 1.27(1.06, 1.57)], and the difference was statistically significant ( P<0.05). When the SWD value >15.60 m·s -1·kHz -1 was used as the cut-off value, the area under ROC curve (AUC) was 0.72, the sensitivity was 88.2%, the specificity was 51.4%, and the accuracy was 75.7%. The sensitivity, specificity, accuracy and AUC were 58.8%, 74.2%, 63.1%, and 0.68, respectively, when the ratio of SWD value<1.32 was used as the cut-off value. SWD value combined with SWD ratio for the diagnosis of liver malignant tumor, the AUC was 0.88, the sensitivity was 82.3%, the specificity was 83.0%, and accuracy was 81.6%. The diagnostic efficacy of the two in combination for liver malignant tumor was superior to SWD value ( Z=2.678, P=0.007 4) and SWD value ratio ( Z=3.822, P=0.000 1). Conclusions:SWD imaging can reflect the viscosity information of liver tumors and surrounding liver parenchyma, and has potential clinical application value in the differentiation of benign and malignant tumors.

Objective:To investigate the contrast-enhanced ultrasound (CEUS) features of inflammatory hepatocellular adenoma (I-HCA).Methods:The contrast-enhanced ultrasound features I-HCA of 28 cases from April 2009 to November 2019 in Zhongshan Hospital, Fudan University were retrospectively analyzed, including arterial phase enhancement pattern, the homogeneity of enhancement, subcapsular enhancement, and the internal perfusion defect. All I-HCA lesions were divided into >5 cm group( n=9) and ≤5 cm group( n=19), the CEUS features between the two groups were compared. Results:All I-HCA lesions were hyper-enhanced in the arterial phase, among which 39.3% (11/28) showed diffuse filling, 39.3%(11/28) showed centripetal filling, and 21.4%(6/28) showed centrifugal filling pattern. Twenty-five percent (7/28) of I-HCAs showed heterogeneous enhancement, 10.7% (3/28) revealed unenhanced areas within the lesions. Subcapsular vessels were observed in 67.7 (21/31) of I-HCA lesions. Heterogeneous enhancement and unenhanced areas were more frequently detected in lesions >5 cm ( P=0.020, 0.026, respectively), while there was no difference in the enhancement pattern and subcapsular vessels between the two groups ( P>0.05). Inportal venous phase, 42.9%(12/28) of I-HCAs showed hypo-enhancement, and 57.1%(16/28) of lesions showed washout in late phase. According to "hyper-enhancement in arterial phase, sustained hyper- or iso-enhancement in portal venous and late phase" by CEUS, the diagnostic accuracy of benign lesion was 42.9%(12/28). According to any of hyper-enhancement pattern in arterial phase, subcapsular vascular enhancement, and sustained hyper- or iso-enhancement in portal venous and late phase, the diagnostic accuracy of I-HCA was 71.4% (20/28). Conclusions:CEUS is valuable in the diagnosis of inflammatory hepatocellular adenoma.

ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.

Objective To investigate the role of combined analysis of E2F3a and CASP8AP2 expression in prognosis evaluation in pediatric acute lymphoblastic leukemia (ALL). Methods The study included 141 newly diag-nosed pediatric ALL patients enrolled at the Hematology Center,Beijing Children′s Hospital,Capital Medical Universi-ty between March 2008 and July 2010,including 97 boys and 44 girls(aged 1. 2 - 15. 5 years,median 5. 2 years). E2F3a and CASP8AP2 expressions were quantified in 141 children with ALL by adopting real - time quantitative poly-merase chain reaction (qPCR). Receiver operating characteristic (ROC)curve was used to find the best cut - off point to divide the patients into E2F3a or CASP8AP2 low - and high - expression groups,and the treatment outcome between the groups was compared. Cox regression was used to analyze the prognostic significance of the combined expression of E2F3a and CASP8AP2. Results The estimated 5 - year relapse free survival(RFS)rate,event free survival(EFS) rate and overall survival (OS)rate of patients with low - E2F3a and low - CASP8AP2 expression were (58. 9 ± 10. 0)％,(56. 0 ± 9. 9)％ and (72. 0 ± 9. 0)％,respectively. They were significantly lower than those of patients with high - E2F3a and high - CASP8AP2 expression,whose RFS,EFS and OS were (94. 9 ± 2. 5)％,(93. 7 ± 2. 7)％ and (96. 2 ± 2. 2)％,and the differences were all statistically significant(all P < 0. 05),respectively. Compared with other patients,the one with low expression of both E2F3a and CASP8AP2 had a poorer prognosis. In addition to MLL rear-rangements and minimal residual disease level at the end of remission induction,low expression of both E2F3a and CASP8AP2 remained as independent prognostic factors. Conclusion Low expressions of E2F3a and CASP8AP2 pre-dict poor prognosis in pediatric ALL. Combined assessment of E2F3a and CASP8AP2 expression could predict poor prognosis and relapse more accurately.

INTRODUCTIONKnowledge of stroke symptoms is associated with seeking medical attention early, and knowledge of risk factors is an essential factor in stroke prevention. In this study, we evaluated the level of stroke literacy in Singapore.

MATERIALS AND METHODSA cross-sectional study of Singapore citizens and permanent residents aged 21 years and above was conducted in a public housing estate. Participants were randomly sampled using multi-stage stratified sampling. Assessment of awareness of stroke symptoms and risk factors was performed using open-ended questions. In total, 687 respondents were recruited, with a response rate of 69.7%.

RESULTSOverall, 52.4% of respondents identified the brain as the source of pathology, and 47.6% could cite at least 1 of the 3 FAST symptoms (facial droop, arm weakness and speech difficulty), while 40% could name 2 or more of 7 established risk factors for stroke (high blood pressure, high cholesterol, cigarette smoking, diabetes mellitus, older age, previous heart attacks and stroke). Respondents at higher risk of stroke (older individuals and those with stroke risk factors) did not have greater awareness of stroke symptoms and risk factors. The majority of respondents reported they would seek immediate medical care if they experienced stroke symptoms. Only 59.4% knew the emergency ambulance service telephone number.

CONCLUSIONIn a sample of Singaporean adults residing in a public housing estate, we found evidence of poor stroke literacy, highlighting the need for comprehensive population-based education efforts. There is a role for opportunistic education among those at higher risk of stroke.

Adult ; Aged ; Cross-Sectional Studies ; Female ; Health Literacy ; Humans ; Male ; Middle Aged ; Public Housing ; Risk Factors ; Singapore ; Socioeconomic Factors ; Stroke ; Surveys and Questionnaires ; Young Adult