OBJECTIVE：To analyze the research status and hotspot frontier of polypharmacy in the elderly in recent ten years ， and to provide reference for polypharmacy study in the elderly in China. METHODS ：The related literatures about polypharmacy in the elderly were retireved from Web of Science core database during Jan. 1st，2010-Dec. 31th，2019. CiteSpace 5.6.R5 software was used to analyze the quantity of pulished literatures ，authors，country/area，institution，co-citation of journals and literatures. The keywords were taken as nodes for co-occurrence analysis ，cluster analysis and Burst analysis ，the visual maps were drawn and the results were analyzed. RESULTS ：A total of 3 521 literatures were included ，and the quantity of published literatures about polypharmacy in the elderly was on the rise ；high-yield authors mainly were Bell JS and Hilmer SN which were the representative with 36 papers published ，accounting for 1.02％ of the total. America （929 literaturesm 26.3％）and University of Sydney （101 literatures，2.87％）were the most high-yield country and institution respectively ；Journal of the American Geriatrics Society was the journal with the most articles （2 202 pieces）. The most frequently cited literature was those published by Radcliff S team in 2015 to update Beers standard to reduce potential inappropriate drug use ；the research on how to optimize the prescription for the elderly published by Spinewine A team in 2007 had the highest co-citation centrality. The hot keywords were elderly ，polypharmacy and risk factor ；keywords with high centrality included quality of life ，polypharmacy and Beers standard. Research hotspots included risk factor ，outcome index and reduction strategy for polypharmacy. Six clusters were formed ，including Breast cancer ， Prescription，Disorder，Schizophrenia，Mediation attachment and Trial ；complications，pain，frailty，health-related quality ， intellectual disability were at the forefront of research. CONCLUSIONS ：In the past 10 years，the research on polypharmacy in the elderly has been paid more and more attention by scholars ；the United States and Australia have made great contributions in this field；the research mainly focused on the impact of polypharmacy on health outcomes and the strategies for reduce polypharmacy. In the future ，more attention will be paid to the related research on psychological ，cognitive and debilitating effects of polypharmacy in the elderly.

Objective:To isolate and purify the chemical constituents from Goodyera schlechtendaliana, and lay a foundation for further research on the effective materials of the plant. Method:The dried grass of G. schlechtendaliana(10 kg)was heated by 95% EtOH for reflux extraction for 4 times at 85℃. The extract liquids were combined, decompressed and concentrated to obtain the crude extract. The crude extract was then suspended in distilled water and extracted with petroleum ether,EtOAc and n-BuOH in turn to obtain corresponding fractions. Silica gel,Sephadex LH-20,ODS and semi-preparation high performance liquid chromatography (HPLC) were used for the separation and purification of EtOAc and n-BuOH fractions. The structures of compounds obtained were identified by ¹H-NMR,¹³C-NMR,MS,physicochemical properties and reference literature. Result:Ten compounds were isolated and identified as 2,5-dihydroxy-4-methoxy-9,10-dihydroxyphenanthrene (1),1,3,5-trimethoxybenzene (2),protocatechuic acid (3),p-hydroxybenzaldehyde (4),p-hydroxybenzoic acid (5),quercetin (6),vanillin(7),daucosterol (8),adenine (9) and adenosine (10). Conclusion:Compounds 1-10 were obtained from the genus of Goodyera for the first time.

BACKGROUND: The dynamic balance between bone formation mediated by osteoblasts and bone resorption mediated by osteoclasts is the basis for maintaining the stability of the body’s bone tissue. The metabolic imbalance between them can cause bone loss and fine structure degeneration of the bone cells, leading to osteoporosis. OBJECTIVE: To review the role of DNA methylation in osteoporosis and to explore the mechanism of exercise affecting DNA methylation and DNA methylation regulating bone metabolism. METHODS: A computer-based search of PubMed and CNKI databases was performed for relevant articles published from January 2002 to April 2020 with “DNA methylation; Osteoporosis; Exercise intervention; Mechanical stress; Osteogenic differentiation” as key words in English and Chinese, respectively. Initially, finally 52 eligible articles were included for result analysis. RESULTS AND CONCLUSION: DNA methylation is a relatively conservative and stable apparent modification, which regulates gene expression, silencing and disease occurrence. Studies have shown that reduced methylation levels of genes such as β-catenin, Runx2, osteopontin (OPG) can promote their expression and activate Wnt Pathway, whereas the reduction of methylation level of Sclerosin, receptor activator of nuclear factor kappa B ligand (RANKL) and other genes can promote their expression, and inhibit Wnt pathway and reduce the ratio of OPG/RANKL, thereby affecting the proliferation, differentiation and function of osteoblasts and osteoclasts, and accordingly regulating dynamic equilibrium between bone formation and bone resorption. Osteoblasts and osteoclasts act as sensitive cells for mechanical stimulation. Bone can transform the mechanical load generated by exercise into biological stimulation that acts on the differentiation and function of related bone cells, thereby regulating bone metabolism. In vitro experiments have indicated that different forms of mechanical stress stimulations can change the methylation level of genes such as OPN and GNAS1 to regulate their expression, which has a positive effect on bone formation. Bone tissue is a mechanically sensitive tissue, and DNA methylation can regulate bone metabolism by regulating a variety of factors.

Autoimmune liver disease is a refractory disease clinically, and there is no particularly effective drug at present. Therefore, it is of important clinical value to develop new effective intervention drugs for the prevention and treatment of autoimmune liver disease. In order to investigate the potential protective effect of artesunate (Art) on concanavalin A (Con A)-induced autoimmune liver injury, different doses of Art (27, 54, 108 mg·kg⁻¹) were orally administered to mice for consecutive 7 days, respectively. Then the Con A was injected into mice via tail vein to induce liver injury models. 8 h after modeling, the mice were sacrificed. The serum and liver tissue were collected for detecting the level of alanine aminotransferase (ALT), and aspartate transaminase (AST), liver pathological histopathology, inflammatory cytokines and nuclear factor (NF-κB) key protein expression level. The results showed that 108 mg·kg⁻¹ Art remarkably reduced Con A-induced liver indexes and serum transaminase levels (ALT and AST) as compared with model group(<0.01). Meanwhile, the liver histopathological changes were obviously alleviated with a significant decrease of pro-inflammatory cytokine including tumor necrosis factor (TNF-α), interferon (IFN-γ), interleukin (IL-6), IL-17 and a higher increase of anti-inflammatory cytokine IL-10 (<0.01). Western blot results showed that 108 mg·kg⁻¹ Art markedly inhibited the expressions of p-p65 and p-IκBα proteins (<0.01). The specific inhibitor of NF-κB, pyrrolidinedithiocarbamate (PDTC) could also significantly inhibit the expressions of p-p65 and p-IκBα with and alleviate liver injuries. Therefore, our results indicated that Art may have a protective action against Con A-induced autoimmune liver injury mainly by suppressing NF-κB signal pathway in mice. The study provides scientific reference for artesunate usage in preventing autoimmune liver injury.

Objective: To analyze the expression differences and clinical significance of galectin 9(Gal-9) in different cervical lesions and human papillomavirus(HPV) type infection. Methods: From May 2016 to April 2018, 42 cases of cervical cancer(CC group), 45 cases of high level cervical intraepithelial neoplasia (HSIL group), 45 cases of lower cervical intraepithelial neoplasia (LSIL group) in Yangzhou Maternal and Child Health-Care Hospital were selected, and 45 cases of normal outpatient medical examination in our hospital were selected as the control group(NC group). Enzyme-linked immunosorbent assay(ELISA) method was used to detect the serum level of Gal-9.The gene amplification technique and the principle of diversion hybridization were used to detect the 37 types of HPV. Results: Of women with cervical lesions, 50 cases(28.3%) were 50 years or more, the CC group had 32 cases (76.2%), HSIL group had 10 cases (22.2%), LSIL group had 8 cases (17.8%), the age of CC group compared with HSIL group and LSIL group had significant differences(t=7.959, 10.440, all P<0.01). The serum level of Gal-9 in CC group [(22.27±1.72)ng/L], HSIL group [(15.43±2.14)ng/L], LSIL group [(15.24±1.89)ng/L] was significantly higher than that in NC group [(12.08±2.13)ng/L](t=16.360, 18.100, 24.440, all P<0.05). The HPV infection rate in patients with cervical lesions reached 84.84%(112/132), the positive rate of HPV16 in CC group was 83.3%(35/42), HSIL group was dominated by HPV16(28.8%), HPV33(20.0%), HPV52(17.7%) and HPV58(13.3%), LSIL group was dominated by HPV52(26.7%), HPV58(24.4%), HPV16(20.0%) and HPV18(8.9%). There was no statistically significant difference in the levels of Gal-9 of HPV16, HPV58, HPV52 and HPV33 patients, indicated that there was no statistically significant correlation between Gal-9 level and the type of HPV infection. Conclusion Early intervention in patients with high HPV infection and Gal-9 levels can help reduce the incidence of cervical cancer.

Objective    To investigate the prevalence and related factors of primary palmar hyperhidrosis in adolescents in Yangzhou. Methods    On-site questionnaire survey was performed on students selected by cluster random sampling from the two colleges and two high or middle schools, with each class as a unit. Data were collected through the questionnaire to make the diagnosis and severity grading. Results    A total of 3 487 copies of the questionnaire were distributed in the survey and 3 299 were finished, among which 3 083 were effective with an effective rate of 88.41%. Among them, 1 358 respondents were males and 1 725 were females; 933 were middle school students, 809 high school students, and the remaining 1 341 college students. According to the diagnostic criteria, 104 respondents were diagnosed with palmar hyperhidrosis with an overall prevalence of 3.37%. There were 60 (4.41%) males and 44 (2.55%) females. Although the prevalence of palmar hyperhidrosis in males was higher than that of females (χ2=8.130，P<0.05), severe palmar hyperhidrosis was more often to be observed in females than in males, and females were also more likely to have hyperhidrosis in other parts of the body. In addition, the age of the first onset of the disease was mainly 10 to 20 years old and 36.54% of the patients had a family history. Conclusion    The prevalence of palmar hyperhidrosis in adolescents in Yangzhou was 3.37%, and there is a significant difference in the gender. The palmar hyperhidros is often accompanied by hyperhidrosis symptoms of other parts of body, and the disease shows an obvious genetic predisposition.

Objective: To explore the relationship between cerebral hemorrhagic transformation (HT) and angiographic early venous filling (EVF) following mechanical thrombectomy for acute ischemic stroke. Methods: A retrospective imaging analysis was performed in the consecutive patients treated from January 2015 to November 2018 for acute anterior circulation large vessel occlusion using mechanical thrombectomy on the Affiliated Hospital of Yangzhou University. The demography, vascular risk factors and other clinical data of the patients were also collected. According to the experimental study of European Cooperative Acute Stroke Study Ⅱ (ECASS Ⅱ), the modified classification of HT after mechanical thrombectomy was divided into HT negative, HT-Ⅰ type and HT-Ⅱ type. The differences in EVF, clinical and demographic characteristics were compared. Multivariate logistic regression analysis was used to identify the independent risk factors for HT and clinical outcome. Diagnostic test characteristics of EVF for HT-Ⅱ type were determined using a receiver operating characteristic curve (ROC) analysis. Results: A total of 98 patients with acute ischemic stroke who received mechanical thrombectomy were enrolled, including HT negative in 48 cases (49.0%, 48/98), HT-Ⅰ in 40 cases (40.8%, 40/98) and HT-Ⅱ in 10 cases (10.2%, 10/98). Significant differences were noted in age, and incidence of atrial fibrillation, EVF and poor outcomes among three groups (P<0.05). Multivariate logistic regression analysis showed that EVF [odds ratio (OR) 5.960, 95%CI 1.750-8.960, P=0.001] and atrial fibrillation (OR 3.485, 95%CI 1.962-18.986, P=0.028) were risk factors for the occurrence of HT-Ⅱ after mechanical thrombectomy. No risk factor for HT-Ⅰ was noted. Baseline National Institute of Health Stroke Scale (NIHSS) score (OR 1.162, 95%CI 1.021-1.345, P=0.038), EVF (OR 5.358, 95%CI 1.665-13.653, P=0.006) and HT-Ⅱ (OR 1.326, 95%CI 1.226-2.038, P=0.032) were independent risk factors for poor outcomes. And the sensitivity and specificity of EVF in prediction for HT-Ⅱ were 80.0% and 86.4% respectively, with the area under the ROC curve of 0.832. Conclusion Presence of EVF after mechanical thrombectomy may be the predictor for HT-Ⅱ, which indicates the poor clinical outcomes for acute ischemic stroke patients.

ObjectiveTo investigate the effect and mechanism of pachymic acid （PA） in Poria on the invasion and metastasis of renal carcinoma cells. MethodThe effect of PA （0， 20， 40， 80， 160 μmol·L^-1） on cell viability was detected by cell counting kit-8（CCK-8）， and the dose of PA was selected for subsequent experiments. The effect of PA （0， 20， 40， 80 μmol·L^-1） on cell proliferation was evaluated by colony formation assay. The effect of PA （0， 20， 40， 80 μmol·L^-1） on cell adhesion ability was observed by cell adhesion assay. The effect of PA （0， 20， 40， and 80 μmol·L^-1） on cell invasion and metastasis was investigated by Wound healing assay and Transwell invasion assay. The inhibitory effect of PA （0， 20， 40， 80 μmol·L^-1） on cell motility was further observed and verified by high-content imaging technology. The effects of PA （0， 20， 40， 80 μmol·L^-1） on the expression of matrix metalloproteinase （MMP）/tissue inhibitor of metalloproteinasas （TIMP） related to invasion and metastasis and Smads were detected by Western blot. ResultCCK-8 results showed that compared with the blank group， the PA groups showed decreased cell viability（P<0.01）， with the half-maximal inhibitory concentration （IC₅₀） of ACHN cells of 70.42 μmol·L^-1 at 24 h. Colony formation assay showed that the number of cell clonal groups in the PA groups was reduced compared with that in the blank group（P<0.01）. Cell adhesion assay showed that compared with the blank group， the PA groups displayed reduced cell adhesion（P<0.01）. Wound healing assay showed that the wound healing rate of cells in the PA groups was lower than that in the blank group （P<0.05，P<0.01）. Transwell invasion assay showed that compared with the blank group， the number of transmembrane cells in PA groups was reduced（P<0.01）. High-content imaging showed that the cumulative migration distance of cells in the PA groups was shorter than that in the blank group（P<0.01）. The results of Western blot showed that the protein expression of MMP-2 and MMP-9 in the PA groups decreased （P<0.01）， and TIMP-1 protein expression increased （P<0.01） compared with those in the blank group. In addition， compared with the blank group， the PA groups showed decreased protein expression of Smad2 and Smad3 （P<0.01）. ConclusionPA can inhibit the invasion and metastasis of renal carcinoma cells presumably through regulating the homeostasis of MMP/TIMP by Smad2/3.

OBJECTIVETo examine the role of Cd-induced reactive oxygen species (ROS) generation in the apoptosis of neuronal cells.

METHODSNeuronal cells (primary rat cerebral cortical neurons and PC12 cells) were incubated with or without Cd post-pretreatment with rapamycin (Rap) or N-acetyl-L-cysteine (NAC). Cell viability was determined by MTT assay, apoptosis was examined using flow cytometry and fluorescence microscopy, and the activation of phosphoinositide 3'-kinase/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and mitochondrial apoptotic pathways were measured by western blotting or immunofluorescence assays.

RESULTSCd-induced activation of Akt/mTOR signaling, including Akt, mTOR, p70 S6 kinase (p70 S6K), and eukaryotic initiation factor 4E binding protein 1 (4E-BP1). Rap, an mTOR inhibitor and NAC, a ROS scavenger, blocked Cd-induced activation of Akt/mTOR signaling and apoptosis of neuronal cells. Furthermore, NAC blocked the decrease of B-cell lymphoma 2/Bcl-2 associated X protein (Bcl-2/Bax) ratio, release of cytochrome c, cleavage of caspase-3 and poly(ADP-ribose) polymerase (PARP), and nuclear translocation of apoptosis-inducing factor (AIF) and endonuclease G (Endo G).

CONCLUSIONCd-induced ROS generation activates Akt/mTOR and mitochondrial pathways, leading to apoptosis of neuronal cells. Our findings suggest that mTOR inhibitors or antioxidants have potential for preventing Cd-induced neurodegenerative diseases.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Caspases ; metabolism ; Mitochondria ; drug effects ; Neurons ; drug effects ; PC12 Cells ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism

【Objective】 To explore the relationship between hidden blood loss （HBL） and risk factors in percutaneous endoscopic interlaminar discectomy （PEID）. 【Methods】 We retrospectively analyzed 113 patients （67 males and 46 females， aged 21-71 years old） with lumbar disc herniation （L5-S1 level） who underwent PEID with complete clinical data in the Department of Spinal Surgery of Subei People’s Hospital Affiliated to Yangzhou University from January 2018 to August 2020. The collected data included general patient indicators （age， body mass index and gender）， underlying diseases （hypertension and diabetes）， laboratory tests （prothrombin time， activated partial thromboplastin time， fibrinogen， hematocrit and hemoglobin level）， imaging related parameters （grade of intervertebral disc degeneration， interlaminar space height and soft tissue thickness of interlaminar approach）， intraoperative significant blood loss and operation time. According to the Gross formula， total blood loss and HBL during operation were calculated. And the risk factors of HBL was analyzed. 【Results】 The total perioperative blood loss was （394.85±130.50） mL， and HBL was （337.09±124.98） mL which accounted for 85.4% of the total blood loss. Multivariate linear regression analysis showed that operation time （P<0.001） and disc degeneration grade （P=0.020） were independent risk factors of HBL. 【Conclusion】 HBL is the main cause of blood loss during PEID， and operation time and disc degeneration grade are independent risk factors that affect the amount of HBL.