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SUMMARY: The objective of this study was to investigate the therapeutic wound healing potential and molecular mechanisms of shikonin as small molecules in vitro. A mouse burn model was used to explore the potential therapeutic effect of shikonin; we traced proliferating cells in vivo to locate the active area of skin cell proliferation. Through the results of conventional pathological staining, we found that shikonin has a good effect on the treatment of burned skin and promoted the normal distribution of skin keratin at the damaged site. At the same time, shikonin also promoted the proliferation of skin cells at the damaged site; importantly, we found a significant increase in the number of fibroblasts at the damaged site treated with shikonin. Most importantly, shikonin promotes fibroblasts to repair skin wounds by regulating the PI3K/AKT signaling pathway. This study shows that shikonin can effectively promote the proliferation of skin cell, and local injection of fibroblasts in burned skin can play a certain therapeutic role.
El objetivo de este trabajo fue investigar el potencial terapéutico de cicatrización de heridas y los mecanismos moleculares de la shikonina como moléculas pequeñas in vitro. Se utilizó un modelo de quemaduras en ratones para explorar el posible efecto terapéutico de la shikonina; Rastreamos las células en proliferación in vivo para localizar el área activa de proliferación de células de la piel. A través de los resultados de la tinción para patología convencional, encontramos que la shikonina tiene un buen efecto en el tratamiento de la piel quemada y promueve la distribución normal de la queratina de la piel en el sitio dañado. Al mismo tiempo, la shikonina también promovió la proliferación de células de la piel en el sitio dañado. Es importante destacar que encontramos un aumento significativo en la cantidad de fibroblastos en el sitio dañado tratado con shikonina. Lo más importante es que la shikonina promueve la función reparadora de fibroblastos en las heridas de la piel regulando la vía de señalización PI3K/ AKT. Este estudio muestra que la shikonina puede promover eficazmente la proliferación de células de la piel y que la inyección local de fibroblastos en la piel quemada puede desempeñar un cierto papel terapéutico.
Subject(s)
Animals , Mice , Wound Healing/drug effects , Burns/drug therapy , Naphthoquinones/administration & dosage , Skin , In Vitro Techniques , Naphthoquinones/pharmacology , Phosphatidylinositol 3-Kinases , Cell Proliferation/drug effects , Disease Models, Animal , Proto-Oncogene Proteins c-akt , Fibroblasts , Mice, Inbred C57BLABSTRACT
Aim To investigate the effect of silencing circRNA ciRS-7 on cisplatin resistance of gastric cancer cells and its related mechanism. Methods Cisplatin-resistant gastric cancer cells HGC-27/DDP were constructed, ciRS-7 was silenced and HGC-27/DDP cells were treated with cisplatin. Real-time quantitative PCR (qRT-PCR) was used to detect the expression levels of ciRS-7 and microrNA-944 (miR-944). Cell counting kit-8 (CCK-8) was used to detect cell survival rate. Clone formation assay was used to detect the number of clones. Western blot was used to detect the protein expression levels of CyclinD1, proliferating cell nuclear antigen (PCNA), B-lymphocytoma-2-associated X protein (Bax), B-lymphocytoma-2 (Bcl-2) and cleaved aspartate-specific cysteine proteinase 3 (cleaved-caspase-3). Flow cytometry was used to detect cell apoptosis rate. In situ end labeling (TUNEL) was used to detect apoptosis index. Dual luciferase assay was used to verify the targeting relationship between ciRS-7 and miR-944. Results CiRS-7 was highly expressed in cisplatin-resistant gastric cancer tissues and cells, while miR-944 was low expressed in cisplatin-resistant gastric cancer tissues and cells. The survival rate, clonal formation number, protein expression levels of CyclinD1, PCNA and Bcl-2 in cisplatin-treated HGC-27/DDP cells were significantly reduced by silencing ciRS-7, while ciRS-7 expression level, apoptosis rate, apoptosis index, Bax, cleaved-caspase-3 protein expression levels of cisplatin-treated HGC-27/DDP cells were significantly raised. CiRS-7 targetedly inhibited the expression of miR-944. Conclusions Silencing ciRS-7 can reverse cisplatin resistance of gastric cancer cells, and the mechanism may be related to the promotion of miR-944 expression by silencing ciRS-7.
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OBJECTIVE@#To investigate the correlation between single-nucleotide polymorphism (SNP) of ARID5B gene and resistance to methotrexate (MTX) in children with acute lymphoblastic leukemia (ALL).@*METHODS@#A total of 144 children with ALL who were treated in General Hospital of Ningxia Medical University from January 2015 to November 2021 were enrolled and divided into MTX resistant group and non-MTX resistant group, with 72 cases in each group. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) technology was used to measure the SNP of ARID5B gene in all children and analyze its correlation with MTX resistant.@*RESULTS@#There were no significant differences in the genotype and gene frequency of rs7923074, rs10821936, rs6479778, and rs2893881 between MTX resistant group and non-MTX resistant group (P>0.05). The frequency of C/C genotype in the MTX resistant group was significantly higher than that in the non-MTX resistant group, while the frequency of T/T genotype was opposite (P<0.05). The frequency of C allele in the MTX resistant group was significantly higher than that in the non-MTX resistant group, while the frequency of T allele was opposite (P<0.05). Multivariate logistic regression analysis showed that ARID5B gene rs4948488 TT genotype and T allele frequency were risk factors for MTX resistant in ALL children (P<0.05).@*CONCLUSION@#The SNP of ARID5B gene is associated with MTX resistant in ALL children.
Subject(s)
Child , Humans , DNA-Binding Proteins/genetics , Gene Frequency , Genotype , Methotrexate , Polymorphism, Single Nucleotide , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Transcription Factors/genetics , Drug Resistance, NeoplasmABSTRACT
Objective To evaluate the effect of the treatment with low-concentration hydrogen peroxide (H2O2) on the adhesive function of and autophagy in human melanocytes,and to screen long noncoding RNAs (lncRNAs) related to autophagy.Methods Melanocytes were isolated from foreskins of healthy males after circumcision,and subjected to cultivation.Melanocytes at exponential growth phase were divided into 3 groups:control group receiving no treatment,H2O2 group treated with 400 μ mol/L H2O2,and H2O2 + NAC group pretreated with 4 mmol/L NAC for 2 hours followed by the treatment with 400 μmol/L H2O2.After 5-day treatment,immunofluorescence study was performed to determine the expression of Ecadherin,microtubule-associated protein 1 light chain 3 (LC3)and p62,and Western blot analysis to determine the expression of autophagy-related protein LC3 and p62.Cell structures and autophagosomes were observed by transmission electron microscopy,and autophagy-related lncRNAs were screened using gene chip technology.Statistical analysis was done with Graphpad Prism 6 software using one-way analysis of variance for comparison among groups,and Tukey's test for multiple comparisons.Results Under the confocal microscopy,the H2O2 group showed significantly decreased fluorescence intensity of E-cadherin and LC3 in the melanocytes and decreased number of autophagosomes in melanocytes,but significantly increased fluorescence intensity of p62 compared with the control group and H2O2 + NAC group.Western blot analysis showed that the LC3-Ⅱ/LC3-Ⅰ ratio in the melanocytes was significantly lower in the H2O2 group (0.604 ± 0.012) than in the control group (1.200 ± 0.081,q =7.718,P ＜ 0.01) and H2O2 + NAC group (1.017 ± 0.062,q =5.076,P ＜ 0.05),while the p62/β-actin ratio in the melanocytes was significantly higher in the H2O2 group (0.881 ± 0.079) than in the control group (0.456 ± 0.121,q =4.847,P ＜ 0.05) and H2O2 + NAC group (0.492 ± 0.049,q =4.439,P ＜ 0.05).There were no significant differences in the LC3-Ⅱ/LC3-Ⅰ ratio or p62/β-actin ratio between the H2O2 + NAC group and control group (P ＞ 0.05).Gene chip technology showed that 18 autophagy-related lncRNAs were associated with premature senescence of melanocytes and differentially expressed in the H2O2 group compared with the control group,and the autophagy-related lncRNA NONHSAT190308.1 (＞ 10-fold increase) was screened out.Conclusion Lowconcentration H2O2 can decrease the expression of E-cadherin and the level of autophagy in melanocytes,and can up-regulate the expression of autophagy-associated lncRNA NONHSAT190308.1.
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<p><b>Background</b>The clinical efficacy of fentanyl for pain control differs greatly across individuals. The purpose of this study was to investigate the impact of CYP3A4*1G polymorphism including wild-type homozygote (CYP3A4*1/*1, GG), mutant heterozygote (CYP3A4*1/*1G, GA), and mutant homozygote (CYP3A4*1G/*1G, AA) on fentanyl analgesia in Chinese patients undergoing hysteroscopy by the assessment of analgesia nociception index (ANI).</p><p><b>Methods</b>A total of 200 gynecologic patients scheduled for elective hysteroscopy under general anesthesia at Peking University People's Hospital from May to December in 2017 were enrolled in this study. Venous blood was withdrawn for genotyping of CYP3A4*1G before operation. Fentanyl 1 μg/kg was administered preoperatively followed by target-controlled infusion of propofol for induction and maintenance. Intraoperative analgesic efficacy of fentanyl was assessed by ANI monitoring at T0 (entering room), T1 (cervical dilation), T2 (start of cervical aspiration), and T3 (end of cervical aspiration) time points. The duration of propofol infusion and total dosage of propofol were recorded as well.</p><p><b>Results</b>The patients were divided into three groups according to CYP3A4*1G polymorphism, including 143 in GG group, 47 in GA group, and 10 in AA group. There was no significant difference in clinical demographics among three groups. The frequency of CYP3A4*1G variant alleles accounted for 16.8% and the distribution of variant alleles was consistent with Hardy-Weinberg equilibrium. Using a multilevel model, ANI values at T1 (63.81 ± 19.61), T2 (63.63 ± 17.82), and T3 (65.68 ± 17.79) were significantly lower than that at T0 (77.16 ± 12.93) in the study population (F = 23.50, P < 0.001), suggesting that higher levels of pain at T1, T2, and T3 than T0. Patients with GG genotype showed significantly lower ANI than those with GA or AA genotypes during hysteroscopy under the same dose of fentanyl.</p><p><b>Conclusion</b>CYP3A4*1G polymorphism associated with the analgesic efficacy of intraoperative fentanyl in the patients undergoing hysteroscopy under general anesthesia.</p>
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Objective To evaluate the effect of hydrogen peroxide (H2O2) on autophagy in melanocytes,and to explore its possible regulatory mechanisms.Methods Normal human melanocytes at exponential growth phase were divided into several groups:blank control group receiving no treatment,positive control group treated with 100 nmol/L sirolimus solution,and experiment groups treated with H2O2 solution at different volume fractions of 10-7-10-3 respectively.After 4-hour treatment,cell counting kit-8 (CCK-8) assay and flow cytometry were performed to evaluate the cellular proliferative activity and detect apoptosis of melanocytes respectively.Acridine orange staining was performed to detect autophagosome formation,transmission electron microscopy to observe ultrastructural changes of autophagosomes,and Western blot analysis to measure the expression of autophagy-specific protein Beclin 1 and microtubuleassociated protein 1 light chain 3B (LC3B).A total of 84 autophagy-related genes were analyzed by RT2 Profiler PCR Array,so as to screen differentially expressed autophagy-related genes.Results After the treatment with H2O2 at different volume fractions of 10-3,5 × 10-4,10-4,5 × 10-5,10-5,5 × 10-6 and 10-6,experiment groups showed significantly decreased cellular proliferative activity,but significantly increased apoptosis rate compared with the blank control group (F =286.95,301.23,respectively,both P ＜ 0.05).With the increase in volume fractions of H2O2,the cellular proliferative activity was significantly gradually decreased (P ＜ 0.05),while the apoptosis rate showed an opposite trend (P ＜ 0.05),except that the 5 ×10-6 H2O2 group showed no significant differences in the apoptosis rate compared with the 10-5 H2O2 group and 10-6 H2O2 group.Acridine orange staining and electron microscopy showed autophagosome formation in the 10-5 H2O2 group,10-6 H2O2 group and positive control group.Western blot analysis revealed that Beclin1 expression and LC3B-Ⅱ/LC3B-Ⅰ ratio were significantly higher in the 10-5 H2O2 group,10-6 H2O2 group and positive control group than in the blank control group (all P ＜ 0.05).RT2 Profiler PCR Array showed significant up-regulation of ATG12,ATG3,ULK1,PIK3CG,PTEN and PIK3C3 genes and significant downregulation of EIF2AK3 gene in the 10-5 H2O2 group,10-6 H2O2 group and positive control group compared with the blank control group.In the 10-5 H2O2 group and positive control group,the mTOR gene was significantly up-regulated,and the ULK2 gene was significantly down-regulated.The 10-6 H2O2 group showed no obvious changes in the expression of mTOR gene,but significant up-regulation of AMPK and JNK1 genes.Conclusion H2O2 at volume fractions of 10-5 and 10-6 can induce autophagy in melanocytes,likely by influencing the expression of some related signaling molecules.
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Objective To investigate the clinical and radiographic features of anterior internuclear ophthalmoplegia caused by acute brainstem infarction. Methods Four cases of anterior internuclear ophthalmoplegia were retrospectively reviewed by analyzing their clinical and radiographic data together with anatomical features. Results All 4 cases were caused by brainstem ischemic lesions, demonstrating dotted infraction on thin-sliced diffused-weighted imaging at the dorsal side of pontine and the ventromedial side of the fourth ventricle. Among 4 cases, 1 was cured and the other 3 cases achieved improved adduction and diplopia. Conclusion Anterior internuclear ophthalmoplegia is characterized by its dotted lesion in the brainstem at the dorsal side of pontine and the ventromedial side of the fourth ventricle. Thin-sliced MRI facilitates the detection of these lesions.
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<p><b>OBJECTIVE</b>To observe the clinical efficacy of penyan pill (PP) in treating ureaplasma urealyticum (UU) infection patients of qi deficiency blood stasis syndrome (QDBSS).</p><p><b>METHODS</b>Totally 188 UU infection patients of QDBSS were randomly assigned to two groups, the treatment group and the control group. Patients in the treatment group were treated with PP (10 g each time, thrice daily, 14 consecutive days as one therapeutic course), while those in the control group took azithromycin (10 g each day, 7 consecutive days as one therapeutic course). They were continually treated for 3 therapeutic courses. The clinical symptom integrals were observed in the two groups before and after treatment. The short-term efficacy was judged. Their recurrence rates were followed-up to assess their long-term efficacies.</p><p><b>RESULTS</b>The total effective rate of the comprehensive efficacy in the treatment group was 91.4%, while it was 79. 3%in the control group, showing no statistical difference between the two groups (P > 0.05). Better effects were obtained in improving Chinese medical clinical symptoms in the treatment group (P <0.01). There was no statistical difference in the negative conversion rate between the two groups after treatment (P >0. 05). There was statistical difference in the recurrence rate between the two groups (12. 82% vs 54.76%,P <0. 05).</p><p><b>CONCLUSIONS</b>PP showed equivalent effects in treating UU infection patients of QDBSS to those of azithromycin. But PP showed obvious advantages over azithromycin in improving Chinese medical syndromes.</p>
Subject(s)
Adult , Female , Humans , Middle Aged , Azithromycin , Therapeutic Uses , Drugs, Chinese Herbal , Therapeutic Uses , Medicine, Chinese Traditional , Phytotherapy , Ureaplasma Infections , Diagnosis , Drug Therapy , Ureaplasma urealyticumABSTRACT
<p><b>OBJECTIVE</b>To develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood.</p><p><b>METHODS</b>A pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616).</p><p><b>RESULTS</b>In the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized.</p><p><b>CONCLUSION</b>This assay has a broad application for clinical diagnosis and surveillance investigation.</p>
Subject(s)
Humans , Base Sequence , Molecular Sequence Data , Reverse Transcriptase Polymerase Chain Reaction , Severe acute respiratory syndrome-related coronavirus , Genetics , Severe Acute Respiratory Syndrome , DiagnosisABSTRACT
Objective To study the protective effect of Astragalus membrance on mice infected with tachyzoites of Toxoplasma gondii. Methods ICR mice were infected intraperitoneally with 10 5, 10 3, 10 2 tachyzoites of virulent RH strain of Toxoplasma gondii, and the mice were orally treated with Astragalus membrance 0 075 g/d per mouse or Azithromycin [150 mg/(d?kg)] each day starting from day 1 post-infection for 10 days. The survival rate and period were investigated. The parasite loads of livers and lungs of the mice infected with 10 2 tachyzoites were determined by fluorescence PCR methods at 4 day-post-infection (dpi) and 8 dpi. Results When infected with 10 5, 10 3 tachyzoites, treated with Astragalus membrance, the average survived days of the mice were 5 57 days and 6 23 days, and treated with azithromycin were 6 96 days and 8 12 days respectively. The azithromycin group but not the astragli group survived significantly longer than the control(P