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To investigate the protective effect and the potential mechanism of leonurine(Leo) against erastin-induced ferroptosis in human renal tubular epithelial cells(HK-2 cells), an in vitro erastin-induced ferroptosis model was constructed to detect the cell viability as well as the expressions of ferroptosis-related indexes and signaling pathway-related proteins. HK-2 cells were cultured in vitro, and the effects of Leo on the viability of HK-2 cells at 10, 20, 40, 60, 80 and 100 μmol·L~(-1) were examined by CCK-8 assay to determine the safe dose range of Leo administration. A ferroptosis cell model was induced by erastin, a common ferroptosis inducer, and the appropriate concentrations were screened. CCK-8 assay was used to detect the effects of Leo(20, 40, 80 μmol·L~(-1)) and positive drug ferrostatin-1(Fer-1, 1, 2 μmol·L~(-1)) on the viability of ferroptosis model cells, and the changes of cell morphology were observed by phase contrast microscopy. Then, the optimal concentration of Leo was obtained by Western blot for nuclear factor erythroid 2-related factor 2(Nrf2) activation, and transmission electron microscope was further used to detect the characteristic microscopic morphological changes during ferroptosis. Flow cytometry was performed to detect reactive oxygen species(ROS), and the level of glutathione(GSH) was measured using a GSH assay kit. The expressions of glutathione peroxidase 4(GPX4), p62, and heme oxygenase 1(HO-1) in each group were quantified by Western blot. RESULTS:: showed that Leo had no side effects on the viability of normal HK-2 cells in the concentration range of 10-100 μmol·L~(-1). The viability of HK-2 cells decreased as the concentration of erastin increased, and 5 μmol·L~(-1) erastin significantly induced ferroptosis in the cells. Compared with the model group, Leo dose-dependently increased cell via-bility and improved cell morphology, and 80 μmol·L~(-1) Leo promoted the translocation of Nrf2 from the cytoplasm to the nucleus. Further studies revealed that Leo remarkably alleviated the characteristic microstructural damage of ferroptosis cells caused by erastin, inhibited the release of intracellular ROS, elevated GSH and GPX4, promoted the nuclear translocation of Nrf2, and significantly upregulated the expression of p62 and HO-1 proteins. In conclusion, Leo exerted a protective effect on erastin-induced ferroptosis in HK-2 cells, which might be associated with its anti-oxidative stress by activating p62/Nrf2/HO-1 signaling pathway.
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Humans , Ferroptosis , Reactive Oxygen Species/metabolism , NF-E2-Related Factor 2/metabolism , Sincalide/pharmacology , Signal Transduction , Epithelial Cells/metabolism , GlutathioneABSTRACT
Objective: To review the clinical data of 7 patients with Danon disease and analyze their clinical characteristics. Methods: The medical records of 7 patients with Danon disease, who were hospitalized in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from April 2008 to July 2021, were reviewed and summarized, of which 6 cases were diagnosed as Danon disease by lysosomal-associated membrane protein-2 (LAMP-2) gene mutation detection and 1 case was diagnosed by clinicopathological features. Clinical manifestations, biochemical indexes, electrocardiogram, echocardiography, skeletal muscle and myocardial biopsy and gene detection results were analyzed, and patients received clinical follow-up after discharge. Results: Six patients were male and average age was (15.4±3.5) years and the average follow-up time was (27.7±17.0) months. The main clinical manifestations were myocardial hypertrophy (6/7), decreased myodynamia (2/7) and poor academic performance (3/7). Electrocardiogram features included pre-excitation syndrome (6/7) and left ventricular hypertrophy (7/7). Echocardiography examination evidenced myocardial hypertrophy (6/7), and left ventricular dilatation and systolic dysfunction during the disease course (1/7). The results of skeletal muscle biopsy in 6 patients were consistent with autophagy vacuolar myopathy. Subendocardial myocardial biopsy was performed in 3 patients, and a large amount of glycogen deposition with autophagosome formation was found in cardiomyocytes. LAMP-2 gene was detected in 6 patients, and missense mutations were found in all these patients. During the follow-up period, implantable cardioverter defibrillator implantation was performed in 1 patient because of high atrioventricular block 4 years after diagnosis, and there was no death or hospitalization for cardiovascular events in the other patients. Conclusion: The main clinical manifestations of Danon disease are cardiomyopathy, myopathy and mental retardation. Pre-excitation syndrome is a common electrocardiographic manifestation. Autophagy vacuoles can be seen in skeletal muscle and myocardial pathological biopsies. LAMP-2 gene mutation analysis is helpful in the diagnose of this disease.
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Adolescent , Child , Female , Humans , Male , Cardiomyopathies/etiology , Glycogen Storage Disease Type IIb/complications , Hypertrophy, Left Ventricular/etiology , Lysosomal-Associated Membrane Protein 2/genetics , Pre-Excitation Syndromes/geneticsABSTRACT
Objective To establish a discriminative dissolution test method for orlistat capsules and evaluate the similarity of dissolution curves of 6 domestic enterprises and imported orlistat capsules.Methods The HPLC method was used,the chromatographic column was Thermo Hyersil GOLD C18(150 mm×4.6 mm,5 μm)column,the mobile phase was acetonitrile and water(85∶15),the flow rate was 1.0 mL·min-1,the detection wavelength was 195 nm,the column temperature was 30℃,and the injection volume was 20 μL.The effects of different concentrations of sodium dodecyl sulfate,different concentrations of sodium chloride and different pH dissolution media on the dissolution curve were investigated,and the best dissolution conditions were selected.The similarity of the dissolution curve was evaluated using the dissolution curve similarity factor method.Results Orlistat had good linear relationship within the range of 5.989-179.697 μg·mL-1(r=0.999 8),and its average recovery rate was 100.4%,with an RSD of 1.1%(n=9).The optimal dissolution conditions selected were as follows:the pH 6.0 phosphate buffer solution containing 1.0%sodium dodecyl sulfate and 0.5%sodium chloride was as dissolution medium;the conduct dissolution tests conducted under conditions of paddle method,75 r·min-1 and medium 1 000 mL.Only one domestic enterprise had a similar dissolution curve between the product and the reference formulation,while the other five enterprises had inconsistent dissolution behavior between the product and the reference formulation.Conclusion This measurement method can effectively distinguish the dissolution behavior of products from different enterprises,and has certain reference significance to evaluate the consistency of the quality and efficacy of orlistat capsules in China.
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The present study analyzed the potential biomarkers of chronic obstructive pulmonary disease(COPD) with lung-Qi deficiency syndrome by non-targeted metabolomics and explored the biological basis of this syndrome. Blood samples of 96 COPD patients with lung-Qi deficiency syndrome(COPD with lung-Qi deficiency syndrome group) and 106 healthy people(healthy control group) were collected, and the metabolic profiles of both groups were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Multivariate statistical analysis and differential metabolite screening were carried out by using Progenesis QI and Simca-P. Metabolic pathways were constructed through the MetaboAnalyst. Seven potential biomarkers, such as L-cystathionine, protoporphyrinogen Ⅸ, and citalopram aldehyde, were identified. Compared with the results in the healthy control group, the content of citalopram aldehyde, N1-methyl-2-pyridone-5-carboxamide, and 11β,17β-dihydroxy-4-androsten-3-one was significantly up-regulated, while that of the other four compounds such as L-cystathionine, dihydrotestosterone, protoporphyrinogen Ⅸ, and D-urobilinogen was down-regulated. These potential biomarkers involved six metabolic pathways, including cysteine and methionine metabolism, porphyrin and chlorophyll metabolism, drug metabolism of cytochrome P450, steroid hormone biosynthesis, glycine, serine, and threonine metabolism, and nicotinate and nicotinamide meta-bolism. This study is expected to provide a certain scientific basis for the research on traditional Chinese medicine syndrome of COPD with lung-Qi deficiency syndrome from the molecular biology level.
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Humans , Aldehydes , Biomarkers , Chromatography, High Pressure Liquid , Citalopram , Cystathionine , Lung , Metabolomics/methods , Pulmonary Disease, Chronic ObstructiveABSTRACT
Parkinson’s disease (PD)is a complex neurodegenerative disorder by motor impairments and non-motor symptoms. While dopamine-based therapies are effective in fighting the symptoms in the early stages of the disease‚ a lack of neuroprotective drugs means that the disease continues to progress. New disease modifying therapies and novel therapeutic strategies are in high demand for PD patients. Genetic studies indicated that both rare and common genetic variants could induce the development PD. As a risk candidate gene for Parkinson’s disease‚ TMEM175 encodes a lysosomal potassium channel protein with new structures‚ and the protein plays an important role in maintaining lysosomal membrane potential and pH stability. With the in-depth understanding for its structure and function‚ TMEM175 deficiency results in decreased lysosomal catalytic activity and the pathological aggregation of α-synuclein. In view of the importance of lysosome potassium channel TMEM175‚ it could be an interesting target for the development of drugs to treat Parkinson’s disease and other neurodegenerative diseases. Herein we review the structure and function TMEM175‚ and focuses on its involvement in the occurrence and development of PD by affecting the function of lysosome as a homeostatic regulator. Future drug screenings based on lysosome TMEM175 may be carried out to maintain the active state or enhance the expression of TMEM175 to improve the condition of PD patients. Further investigations are needed to study how to maintain the balance between the open and closed state of TMEM175 channels to regulate the ion homeostasis of lysosomes. Studies of this ion channel protein will bring new strategies and ideas for the treatment of PD‚ and provide support for establishing the molecular status of TMEM175 in the diagnosis and treatment of PD.
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Lactic acid bacteria (LAB) are a representative probiotic. As the dominant flora in the human intestinal tract, LAB can regulate the balance of human intestinal flora and improve host health. The purpose of this study was to isolate and screen LAB that are well suited to the intestinal characteristics of the Chinese population, with excellent probiotics and high antibacterial activity. After 16S ribosomal RNA (rRNA) homology and phylogenetic tree analysis, potential probiotics were tested for their antibacterial activity, resistance to artificial gastrointestinal fluid and drugs, surface hydrophobicity, and safety. Three strains of LAB with acid resistance, bile salt resistance, epithelial cell adhesion, and no multidrug resistance were selected:
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objective:To observe the effect of high-flow nasal cannula (HFNC) oxygen therapy on patients with acute respiratory failure after gastrointestinal operation under general anesthesia and its efficacy on prognosis.Methods:Totally 107 patients with acute respiratory failure after gastrointestinal operation under general anesthesia in ICU of our hospital were selected from January 2017 to June 2019. Patients were randomly divided into the HFNC group ( n=57) and non-invasive ventilation (NIV) group ( n=50). The changes of pH, PaO 2, PaCO 2, PaO 2/FiO 2, SpO 2, heart rate (HR), respiration rate (RR), and intra-abdominal pressure (IAP) before and after oxygen treatment were compared. The differences of comfort level, duration of oxygen treatment, re-intubation rate, 48-h pulmonary infection rate, incidence of anastomotic fistula, length of stay in ICU, length of hospital stay, hospitalization cost, and 28-day mortality were compared between the two groups. The t test of two independent samples was used for the comparison of normal measurement data. Non-normal data were expressed by median (quartile), fractional count data by case number (percentage), and comparison between the two groups by Chi-square test. Results:PaO 2, PaCO 2, PaO 2/FiO 2 and SpO 2 were not significantly different after 2-h oxygen therapy. PH of the HFNC group was lower than that of the NIV group (7.39 ± 0.04 vs. 7.42 ± 0.03), PaO 2, PaCO 2, PaO 2/FiO 2 and SpO 2 were higher than that of the NIV group [ (89.22 ± 8.70) vs. (84.99± 9.76) mmHg, (41.3 ± 3.43) vs. (39.34 ± 4.21) mmHg, (250.07 ± 18.34) vs. (237.89±19.38) mmHg, (96.14 ± 2.19) vs. (94.78 ± 2.76)%, P <0.05]; pH, PaO 2, PaCO 2, PaO 2 /FiO 2 and SpO 2 were significantly higher in the HFNC group than those in the NIV group [ (7.39 ± 0.04) vs. (7.36 ± 0.04) ; (97.2 ± 12.45) vs. (93.82 ± 12.54) mmHg; (40.84 ± 5.22) vs. (45.10 ± 6.40) mmHg; (277.16 ± 13.98) vs. (248.86 ± 12.81) mmHg, (98.14 ± 1.64)% vs. (95.48 ± 2.71) %] after 12 h oxygen treatment. The HR, RR and IAP of the HFNC group were lower than those of the NIV group, and the differences were statistically significant ( P <0.05). The duration of oxygen therapy in the HFNC group was shorter than that in the NIV group [(32.01 ± 7.57) vs. (40.88 ± 8.89) h], the reintubation rate was lower than that in the NIV group (1.75% vs. 12%), the pulmonary infection rate within 48 h oxygen therapy was lower than that in the NIV group (8.78% vs.30%), and the length of stay in ICU was shorter than that in the NIV group [(5.61 ± 1.73) vs. (7.60 ± 2.31) d], and the hospitalization cost was lower than that in the NIV group ( t = 4.822, P <0. 05). Conclusions:HFNC can improve the oxygenation index of patients with hypoxemia after gastrointestinal operation under general anesthesia, reduce oxygen treatment time, reduced reintubation rate, reduce pulmonary infection rate within 48 h, and improve the prognosis.
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Tumor immune therapy has been remarkably successful in recent years and several kinds of PD-1/PD-L1 (programmed death-1/programmed death-ligand 1) antibody drugs have been approved by the FDA for treatment of advanced malignant neoplasms. However, as biomacromolecules these antibody drugs have certain drawbacks such as high cost, injection-only administration and immunogenicity; thus, we turned to small molecules that have lower immune risks and better modifiability. Considering the structural diversity of natural products, we chose to investigate the active components in Panax ginseng, a famous and highly valued traditional Chinese medicine. Nine compounds were separated and identified in this research using a HPLC-coupled MS system, and 3 PD-1 binding compounds were identified using the SPR method. The PD-1/PD-L1 inhibitory ability of ginsenoside Rg1, as a representative ginsenoside, was verified by cytopharmacological methods. This research provides a new method for the identification of immune blockade inhibitors in natural products.
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Objective: To establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus,analyze the content of total polysaccharides in samples with different sources and grades. Method: The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system.Dextran with similar molecular weight as polysaccharides was selected as the reference substance.Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus.Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630 nm. Result: The linearity,stability,precision,repeatability and recovery rate of the established method all reached the standards,respectively.The content of total polysaccharides in samples from different areas ranged from 0.87% to 1.39%.The content of total polysaccharides in samples with different grades was 1.40% for first-grade pieces,1.21% for second-grade pieces, and 1.03% for third-grade pieces. Conclusion: The established method is simple,accurate and reproducible,and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly.The content of polysaccharides in samples with different grades shows a certain regularity.The content of polysaccharides is the highest in the first-grade pieces,followed by the content in the second-grade,and the lowest in the third-grade.The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.
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@#【Objective】To screen survival-related differential expression of long non-coding RNA(lncRNA)and its co-expressed genes in breast cancer patients and to verify their expression in breast cancer cells.【Methods】RNA-seq data of 943 cases(837 breast cancer + 106 normal controls)by the TCGA database were screened,and found that long non-coding MAPT-AS1 highly expressed,and breast cancer patients had longer survival. The long non-coding MAPT- AS1 overexpression and interference plasmid was constructed,and the constructed plasmid was transfected into breast cancer cell line T47D,and the stably expressed T47D cell line was screened by puromycin. The expression of long non-coding MAPT-AS1 and its co-expressed genes was verified by the methods of RT-qPCR.【Results】Fluorescence microscopy and RT-qPCR confirmed that the long non-coding MAPT-AS1 overexpression and interference-transfected breast cancer cell line T47D were successfully constructed,and the long non-coding MATS-AS1 interference fragment shRNA3 with the highest interference efficiency was screened. The expression of MAPT ,MAPT- IT1 and NXNL2 in the co-expressed gene was decreased after transfection of the shRNA3 interference fragment ,which was consistent with the expression trend of the long non-coding MAPT-AS1.【Conclusion】The long non-coding MAPT-AS1 overexpression and interference plasmid transfected breast cancer cell line T47D were successfully constructed,and the expression of the co- expressed gene was consistent with the database. The study laid the foundation for further study of the mechanism of action of long non-coding MAPT-AS1 gene in breast cancer.
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To evaluate the therapeutic effect of traditional Chinese medicine( TCM) retention enema in treating ulcerative colitis( UC) by Meta-analysis method. Randomized controlled trials( RCTs) of TCM retention enema in treatment of UC were retrieved from databases as CNKI,Wan Fang,CBM,VIP and PubMed from inception to June 2019. The quality of RCTs was assessed by using the Cochrane collaboration's tool for assessing risk of bias,Meta-analysis were performed with Rev Man 5. 3 software and publication bias was tested by using Stata 15. 1 software. There were twenty-eight articles enrolled,and 2 477 patients were included. The result of Meta-analysis showed that retention enema with TCM had significantly better effectiveness in overall curative effect( RRSASP= 1. 18,95%CI[1. 13,1. 22],Z = 8. 32,P < 0. 01; RR5-ASA= 1. 13,95% CI[1. 03,1. 21],Z = 2. 61,P < 0. 01) symptom curative effect( RR =1. 44,95%CI[1. 22,1. 71],Z = 4. 25,P<0. 01) than those of the control group,and the treatment group was lower than the control group in terms of recurrence( RR = 0. 31,95% CI[0. 17,0. 56],Z = 3. 88,P< 0. 01) and adverse events( RR = 0. 38,95% CI[0. 18,0. 78],Z = 2. 64,P<0. 01),with statistically significant differences. However,there was no significant difference in Meta-analysis result of colonoscopic mucosal change between the two groups. TCM enema is an effective method to treat ulcerative colitis.
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Humans , Male , Colitis, Ulcerative , Drugs, Chinese Herbal , Enema , Medicine, Chinese TraditionalABSTRACT
Background@#Betel quid chewing has been a major risk factor for oral cancer (OC) in southern China. This study aimed to analyze the scientific publications on the relationship between betel quid chewing and OC and construct a model to quantitatively and qualitatively evaluate pertinent publications from 1998 to 2017.@*Methods@#The publications from 1998 to 2017 were retrieved from the Web of Science Core Collection database. Microsoft Excel, Thomson Data Analyzer, VOSviewer, and CiteSpace software were used to analyze the publication outcomes, journals, countries/regions, institutions, authors, research areas, and research frontiers.@*Results@#A total of 788 publications on the relationship between betel quid chewing and OC published until October 25, 2017, were identified. The top 4 related journals were Journal of Oral Pathology Medicine, Oral Oncology, Plos One, and International Journal of Cancer. The top five countries engaged in related research included China, India, the United States, the United Kingdom, and Malaysia. The corresponding disciplines, such as oncology, oral surgery, pathology, environmental and occupational health, and toxicology, were mainly concentrated in three disciplines. The subject terms squamous cell carcinoma, OC, betel quid, expression, oral submucous fibrosis, India, and p53 ranked first among research hotspots. The burst terms squamous cell carcinoma, OC, betel quid, and expression ranked first in research frontiers.@*Conclusions@#Research in this area emphasized hotspots such as squamous cell carcinoma, OC, oral submucosal fibrosis, betel quid, and tobacco. The annual number of publications steadily decreased from 1998 to 2017, with a lack of a systematic study from interdisciplinary perspectives, inadequate pertinent journals, limited regions with the practice of betel quid chewing, and insufficient participation of researchers, which indicate that as the prevalence of OC increases, particularly in China, research in this area warrants further expansion.
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Humans , Areca , Bibliometrics , China , Epidemiology , Malaysia , Epidemiology , Mouth Neoplasms , Epidemiology , Risk Factors , United Kingdom , EpidemiologyABSTRACT
The particularity of healthcare markets made the pricing of medical services a contentious issue.It reviewed the characteristics,technical and institutional arrangements for the outpatient service price setting across Organization for Economic Co-operation and Development(OECD) countries,aiming at providing international experiences for medical service pricing reform in China.
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Objective To investigate the prevalence of primary teeth caries and oral hygiene status of preschool children in Kunming.Methods 6958 children aged 3-5 years from 30 kindergartens in Kunming were randomly selected for this study.The reserch of caries prevalence rate and soft dirt were investigated.Results The caries prevalence rate and the mean decayed,missing and filled teeth (XDMFT) values in primary teeth were 58.68%and 2.61 respectively.We can see the difference of significant between Dental caries prevalence and mean debris index simplified ( DI - S) scores between age groups, and there was no statistical significance of the same index between sex. There was no correlation between the investigation of dental caries and oral hygiene status. Conclusion The caries prevalence rate in 3-5 year-old children in the downtown area of Kunming city is very high,pointing out that preventive treatment against primary teeth caries should be strengthened.
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Objective To investigate the frequency and location of cerebral microbleeds in cerebral autosomal dominant arteriopathy with subcortical infarct and leukoencephalopathy (CADASIL) to understand the imaging and clinical features of the disease.Methods Cranial magnetic resonance imaging and susceptibility-weighted imaging were assessed in seven symptomatic CADASIL patients in People's Hospital of Zhengzhou University from 2014 to 2017.Imaging features and clinical significance of these patients were analyzed retrospectively.Results The seven patients were diagnosed by Notch3 gene detection.Mutations were found in exon 11 in four cases,and in exon 4 in three cases.All the seven patients with CADASIL had cerebral microbleeds,the number of which was 108 (4-36).The number of cerebral microbleeds was found to be higher in cortico-subcortical region than in any other regions.One of CADASIL patients with cerebral microbleeds had intracerebral hemorrhage located in external capsule.The patient with intracerebral hemorrhage had hypertension and multiple cerebral microbleeds.Conclusions Cerebral microbleeds are common imaging characteristics in symptomatic CADASIL,most of which locate in cortico-subcortical region.Cerebral hemorrhage is one of the clinical manifestations of CADASIL patients.
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The aim of this study was to investigate the expression of macrophage migration inhibitory factor (MMIF),hypoxia-inducible factor-1 α (HIF-1 αt) and vascular endothelial growth factor (VEGF) in the serum and endometrial tissues of patients with endometriosis (EM) and the clinical significance.Eighty EM patients [American Reproductive Association stage I (n=20),stage Ⅱ (n=22),stage Ⅲ (n=21) and stage Ⅳ (n=17)] were enrolled and divided into mild (10-14 points,n=28),moderate (16-24 points,n=27) and severe (26-30 points,n=25) dysmenorrhea groups.The control group included 40 healthy women of childbearing age who underwent routine healthcare examinations in the enrolment period.The expression of MMIF,HIF-1α and VEGF in the serum and endometrial tissues was measured by enzyme-linked immunosorbent assay and Western blotting,respectively.Meanwhile,the sensitivity and specificity of serum MMIF,HIF-1α,and VEGF when separately used as single indexes or jointly used as one index were examined as well.The results showed that serum concentrations of MMIF,HIF-1α,and VEGF were significantly higher in EM patients than in controls (P<0.05).The expression of all three proteins in both serum and endometrial tissues increased significantly with the R-AFS stage (P<0.05) and with dysmenorrheal severity (P<0.05).The sensitivity and specificity of the combined detection of serum MMIF,HIF-1α,and VEGF levels were significantly higher than those of single index detection (P<0.05).In conclusion,the expression of MMIF,HIF-1α,and VEGF in the serum and endometrial tissues may be used to assess the stage of EM and the severity of dysmenorrhea.Combined evaluation of MMIF,HIF-1α,and VEGF significantly improves the diagnostic sensitivity and specificity.
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The aim of this study was to investigate the expression of macrophage migration inhibitory factor (MMIF),hypoxia-inducible factor-1 α (HIF-1 αt) and vascular endothelial growth factor (VEGF) in the serum and endometrial tissues of patients with endometriosis (EM) and the clinical significance.Eighty EM patients [American Reproductive Association stage I (n=20),stage Ⅱ (n=22),stage Ⅲ (n=21) and stage Ⅳ (n=17)] were enrolled and divided into mild (10-14 points,n=28),moderate (16-24 points,n=27) and severe (26-30 points,n=25) dysmenorrhea groups.The control group included 40 healthy women of childbearing age who underwent routine healthcare examinations in the enrolment period.The expression of MMIF,HIF-1α and VEGF in the serum and endometrial tissues was measured by enzyme-linked immunosorbent assay and Western blotting,respectively.Meanwhile,the sensitivity and specificity of serum MMIF,HIF-1α,and VEGF when separately used as single indexes or jointly used as one index were examined as well.The results showed that serum concentrations of MMIF,HIF-1α,and VEGF were significantly higher in EM patients than in controls (P<0.05).The expression of all three proteins in both serum and endometrial tissues increased significantly with the R-AFS stage (P<0.05) and with dysmenorrheal severity (P<0.05).The sensitivity and specificity of the combined detection of serum MMIF,HIF-1α,and VEGF levels were significantly higher than those of single index detection (P<0.05).In conclusion,the expression of MMIF,HIF-1α,and VEGF in the serum and endometrial tissues may be used to assess the stage of EM and the severity of dysmenorrhea.Combined evaluation of MMIF,HIF-1α,and VEGF significantly improves the diagnostic sensitivity and specificity.
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Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell (SKOV3). Methods:Cell proliferation was detected by MTT method. The cell apoptosis was detected by FCM Annexin V/PI double staining and transmission electron microscopy. The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR. The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR. Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0. 05) in dose-and time-de-pendent manner. When combined with cisplatin, berberine showed synergistic anticancer effects. Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene. In addition, berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA. Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.
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AIM:To investigate whether Yiqi-Wenyang-Huoxue-Huatan formula ( YWHHF) attenuats brain injury induced by hypoxia-hypercapnia pulmonary hypertension ( HHPH) in the rats by inhibiting excessive endoplasmic re-ticulum stress response .METHODS:Healthy SPF male SD rats ( n=50 ) were randomly divided into 5 groups: control group, hypoxia-hypercapnia group, low-dose YWHHF group, middle-dose YWHHF group and high-dose YWHHF group. The rats in control group lived in normal environment , while the rats in other 4 groups were raised for 4 weeks in oxygen tank with low oxygen concentration and high CO 2 concentration .YWHHF was perfused in the rats of low-, middle-and high-dose groups at 0.15, 0.3 and 0.6 g/kg daily, respectively.The rats in hypoxia-hypercapnia group were given isomet-ric distilled water.The surgery was performed on the rats after 4 weeks, and the brain and lung tissues were quickly collect-ed to detect brain water content and observe the morphological changes after mean pulmonary artery pressure recording and heart perfusion .The caspase-3 activity and the apoptotic index of the brain cells were determined .The expression of c-Jun N-terminal kinase (JNK), caspase-12, C/EBP homologous protein (CHOP) and glucose-regulated protein 78 (GRP78) at protein and mRNA levels in brain tissues was detected by Western blot and RT-PCR.RESULTS:Compared with con-trol group, mean pulmonary artery pressure, brain water content, brain apoptotic index, caspase-3 activity, and the protein and mRNA levels of JNK, caspase-12, CHOP and GRP78 in the rest 4 groups were increased , and the brain and lung tis-sues had obvious damage under light microscope .Compared with hypoxia-hypercapnia group , mean pulmonary artery pres-sure, brain water content , brain apoptotic index , caspase-3 activity, and the protein and mRNA expression of JNK , caspase-12, CHOP and GRP78 in low-, middle-and high-dose YWHHF groups were decreased , and the pathological dam-age of the brain and lung tissues was obviously reduced under light microscope .These changes in middle-dose YWHHF group were the most significant .CONCLUSION:YWHHF effectively relieves the brain injury induced by HHPH in rats , which may be associated with inhibiting excessive endoplasmic reticulum stress response .
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Objective To establish a hanging drop 3D cell culture model of human colon cancer cell (HT29) in 48-well cell culture plate,at the same time,through the comparison of several cell viability detection methods to determine the appropriate one for this cell culture way.Methods HT29 cells of 2 375,3 164,4 218,5 625,7 500 and 10 000/well were seeded in the bottom of the 48-well culture plate to form droplets.After 2 d of inversion culture,the cell spheroids were formed and incubated in medium for another 3 d.The volume of cell spheroids were measured,and the absorbance (A) values were detected through APH assay,MTT assay,MTT assay after digestion,CCK-8 assay and CCK-8 assay after digestion.The results were compared among different methods.Results After 5 d of culture,the cell spheroids were formed perfectly at the density of 2 375-10 000/well,and the volumes were in good linear with the original cell inoculation number at the density of 2 375-7 500/well.The A values of APH assay,MTT assay after digestion and CCK-8 assay after digestion increased with the increase of cell inoculation amount;But the cell ball digestion process was complex,and the cell viability was damaged.However,the A values of MTT and CCK-8 assay increased slowly.Conclusion The method of a hanging drop 3D cell culture model in 48-well culture plate combining with APH assay to detect cell viability is economical,accurate and easy to operate.