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Prostate cancer is now the second most common malignancy in men worldwide, with an increasing incidence in China. Most prostate cancer patients receive whole-gland therapy after diagnosis, but patients with localized prostate cancer may not benefit from the treatment due to side effects. With the development of imaging technology and the theory of "index lesion," focal therapy has been greatly developed, which includs high intensity focused ultrasound, focal laser ablation, cryotherapy, irreversible electroporation and photodynamic therapy. This study reviews the clinical trials in recent years and reveals that high intensity focused ultrasound and focal laser ablation have better failure-free survival and postoperative functional control compared with other focal therapy techniques.
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ObjectiveTo explore the mechanism of aerobic capacity on depression in school-age children, and the multiple mediators of the five dimensions of psychosocial functioning (emotional symptoms, conduct problems, peer problems, prosocial behavior and hyperactivity) between aerobic capacity and depression. MethodsFrom October to December, 2021, pupils of Grade two to Grade five from two primary schools were chester-sampled and investigated using 20-meter multistage shuttle run test, Depression Self-Rating Scale for Children, Self-reported Strengths and Difficulties Questionnaire. ResultsA total of 391 pupils underwent 20-meter multistage shuttle run test, and 312 out of them answering the questionnaires, and 294 questionnaires were valid. Aerobic capacity, depression, emotional symptoms, peer problems, prosocial behavior and hyperactivity were significantly correlated with each other (|r| > 0.127, P < 0.05) (except aerobic capacity and peer problems, and emotional symptoms and prosocial behavior). The results of the multiple mediation effect model showed that aerobic capacity could directly and negatively predict depression, and the mediating effects of emotional symptoms, peer problems, prosocial behavior and hyperactivity were significant, accounting for 34.37%, 12.54%, 34.06% and 17.80% of the total mediating effect, respectively. ConclusionThe aerobic capacity could not only directly affect depression of school-age children, but also improve their psychosocial functioning by reducing emotional symptoms, peer problems and hyperactivity, and increasing prosocial behavior, to indirectly affect their depression.
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Brain edema belongs to the category of "stroke" and "true headache", while Traditional Chinese Medicine mostly understands its core disease mechanisms from the perspectives of stasis, deficiency, and heat, and mostly treats the disease by using warming yang to induce diuresis and eliminating stasis to remove water. Wuling Powder has been lauded as the "first party to typhoid and relieving diuresis", which is used to cure clearing damp and promoting diuresis and warming yang and transforming qi, and has been clinically used in the treatment of brain edema caused by various causes such as head trauma, intracerebral hemorrhage, cerebral infarction, and intracranial space occupying, all with remarkable efficacy. Wuling Powder improves cellular energy supply, scavenges excess oxygen radicals and calcium ions in brain tissue, and reduces the damage to brain tissue caused by vascular inflammatory factors and regulates aquaporins and vascular endothelial growth factor, thereby achieving therapeutic effects.
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Cancer stem cell-like cells (CSCs) play an integral role in the heterogeneity, metastasis, and treatment resistance of head and neck squamous cell carcinoma (HNSCC) due to their high tumor initiation capacity and plasticity. Here, we identified a candidate gene named LIMP-2 as a novel therapeutic target regulating HNSCC progression and CSC properties. The high expression of LIMP-2 in HNSCC patients suggested a poor prognosis and potential immunotherapy resistance. Functionally, LIMP-2 can facilitate autolysosome formation to promote autophagic flux. LIMP-2 knockdown inhibits autophagic flux and reduces the tumorigenic ability of HNSCC. Further mechanistic studies suggest that enhanced autophagy helps HNSCC maintain stemness and promotes degradation of GSK3β, which in turn facilitates nuclear translocation of β-catenin and transcription of downstream target genes. In conclusion, this study reveals LIMP-2 as a novel prospective therapeutic target for HNSCC and provides evidence for a link between autophagy, CSC, and immunotherapy resistance.
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Humans , Autophagy , Carcinoma, Squamous Cell/pathology , Cell Line, Tumor , Glycogen Synthase Kinase 3 beta/metabolism , Head and Neck Neoplasms/pathology , Neoplastic Stem Cells/pathology , Squamous Cell Carcinoma of Head and Neck/pathology , Lysosomal Membrane ProteinsABSTRACT
Objective:To determine the expression of microRNA-188-5p (miR-188-5p) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to assess the effect of its downregulation on the proliferation and invasion of CSCC cells.Methods:From November 2012 to October 2018, 50 surgically resected CSCC tissue specimens and 50 paracancerous normal skin tissue specimens were collected from the First Affiliated Hospital of Xinxiang Medical College in Henan Province. Real-time fluorescence-based quantitative PCR (qPCR) was employed to determine the expression of miR-188-5p in CSCC tissues, paracancerous normal skin tissues, CSCC cell lines SCL-1, A431 and HSC-5, and a human immortalized keratinocyte line HaCaT. Cultured A431 and HSC-5 cells were both divided into 2 groups: miR-188-5p inhibitor group and negative control group, which were transfected with a miR-188-5p inhibitor and its negative control respectively. Then, qPCR was performed to determine the relative expression level of miR-188-5p (expressed as 2 -△△Ct), and cell counting kit-8 (CCK8) and Transwell assays were conducted to assess cellular proliferative activity and invasive ability respectively in the above groups. Dual-luciferase reporter assay was performed to investigate interactions between miR-188-5p and phosphatase and tensin homologue deleted on chromosome 10 (PTEN), and Western blot analysis to determine the protein expression of PTEN, total Akt (t-Akt) and phosphorylated Akt (p-Akt). Two independent samples were compared by using t test. Results:The relative expression level of miR-188-5p was significantly higher in the CSCC tissues (5.213 ± 3.138) than in the paracancerous normal skin tissues (1.010 ± 0.364, t = 9.187, P < 0.001), and significantly higher in the SCL-1, A431 and HSC-5 cells (3.858 ± 0.163, 7.068 ± 0.262 and 4.572 ± 0.413, respectively) than in the HaCaT cells (1.079 ± 0.300, t = 17.890, 21.110 and 8.737, respectively, all P < 0.05). Compared with the negative control group, the miR-188-5p inhibitor group showed significantly decreased miR-188-5p expression in both A431 and HSC-5 cells (both P < 0.01), and decreased proliferative activity and invasive ability of both A431 and HSC-5 cells (all P < 0.05). Dual-luciferase reporter assay showed that the downregulation of miR-188-5p significantly increased the expression of PTEN, but inhibited the expression of p-Akt in A431 and HSC-5 cells. Conclusion:MiR-188-5p is highly expressed in CSCC tissues and cells, and the downregulation of miR-188-5p may inhibit the proliferative activity and invasive ability of CSCC cells by regulating the PTEN/Akt pathway.
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Objective:To assess the translation activity of circ-PTPN22, and to investigate its relationship with systemic lupus erythematosus (SLE) .Methods:From May 10, 2019 to September 30, 2019, whole blood samples were collected from 6 female patients with SLE and 9 healthy female controls in the Department of Rheumatology and Immunology, Integrated Traditional Chinese and Western Medicine, and Physical Examination Center of Southwest Hospital, respectively. Peripheral blood mononuclear cells (PBMCs) were isolated from these blood samples, and PBMCs from 3 cases of SLE and 3 healthy controls were sorted into T, B and NK cells by using magnetic beads. The circ-PTPN22 internal ribosomal entry site (IRES) sequence and protein sequence translated from it were predicted in the circRNADb database, rabbit anti-circ-PTPN22pro polyclonal antibodies were prepared against the specific amino acid sequence at the circ-PTPN22 splice site, and Western blot analysis was performed to determine the protein expression of circ-PTPN22pro in PBMCs and T, B and NK cell subsets of the healthy controls and patients with SLE. Cultured Jurkat cells were divided into 4 groups to be transfected with recombinant lentiviral vectors carrying circ-PTPN22-FLAG, circ-PTPN22-NC-FLAG, circ-PTPN22-shRNA-FLAG, circ-PTPN22-shRNA-NC-FLAG respectively, with the normally cultured cells as the cell control group. Then, Western blot analysis was performed to determine the protein expression of circ-PTPN22pro in Jurkat cells, flow cytometry to evaluate the effect of circ-PTPN22 on cell activation and apoptosis. Statistical analysis was carried out by using two-way repeated measures analysis of variance and two-independent-sample t test. Results:Based on the circRNADb database, circ-PTPN22 was predicted to have a translation ability, and Western blot analysis showed that the relative molecular mass of circ-PTPN22pro was 20 000. Forty-eight hours after transfection, circ-PTPN22pro expression was significantly higher in the circ-PTPN22-FLAG group than in the circ-PTPN22-NC-FLAG group and cell control group. At 24, 48 and 72 hours after transfection, the interleukin 2 (IL-2) expression was significantly lower in the circ-PTPN22 group (22.20% ± 8.92%, 31.10% ± 5.88%, 53.20% ± 10.25%, respectively) than in the circ-PTPN22-NC Group (30.90% ± 11.00%, 51.23% ± 10.70%, 69.67% ± 9.00%, respectively; F = 284.881, P = 0.003) , but significantly higher in the circ-PTPN22-shRNA group (35.57% ± 8.79%, 78.10% ± 10.08%, 88.63% ± 3.89%, respectively) than in the circ-PTPN22-shRNA-NC group (26.73% ± 4.92%, 41.03% ± 10.45%, 41.33% ± 4.96%, respectively; F = 293.818, P = 0.003) . After 48, 72 and 96 hours after transfection, the apoptosis rate was significantly higher in the circ-PTPN22 group than in the circ-PTPN22-NC group ( F = 81.287, P = 0.012) , as well as in the circ-PTPN22-shRNA group than in the circ-PTPN22-shRNA-NC group ( F = 111.813, P = 0.009) . The SLE group showed decreased (almost no) circ-PTPN22pro expression in PBMCs compared with the healthy control group. The circ-PTPN22pro expression in T and B cells was significantly lower in the SLE group than in the healthy control group ( t = 3.047, 4.806, both P <0.05) , and there was no significant difference in the circ-PTPN22pro expression in NK cells between the two groups ( t = 0.582, P > 0.05) . Conclusions:Circ-PTPN22 can be translated into circ-PTPN22pro protein, and can inhibit the activation of Jurkat cells. The circ-PTPN22pro expression is lower in PBMCs of the SLE patients than in those of the healthy controls, suggesting that circ-PTPN22 may be related to the occurrence and development of SLE.
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Objective:To evaluate associations of psoriasis with adverse pregnancy outcomes.Methods:Databases, including CNKI, Wanfang, VIP, PubMed, Embase and Cohrane Library databases, were searched for published articles on associations between psoriasis and adverse pregnancy outcomes between January 1980 and December 2018. Quality of included articles was assessed by using MOOSE checklist. Statistical analysis was carried out with Review Manager 5.3 software.Results:One cohort study, 4 case-control studies and 2 cross-sectional studies meet the inclusion criteria. After heterogeneity test, meta-analysis was carried out by using a random effect model. The risks of cesarean ( OR= 1.17, 95% CI: 1.01- 1.37) , eclampsia or preeclampsia ( OR= 1.34, 95% CI: 1.00- 1.79) and premature birth ( OR= 1.09, 95% CI: 1.00- 1.19) were significantly higher in patients with psoriasis than in individuals without psoriasis (all P < 0.05) . There was no significant difference between patients with psoriasis and individuals without psoriasis in the risks of spontaneous abortion, low birth weight, high birth weight, stillbirth, congenital malformation, placental abruption, overdue delivery, low Apgar score (< 7) , polyhydramnios and oligohydramnios. Conclusion:The risks of cesarean, eclampsia or preeclampsia, and premature birth are higher in patients with psoriasis than in individuals without psoriasis.
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Human adenovirus is one of the most important pathogen causing acute respiratory infections in infants and young children, and is easy to cause severe adenovirus pneumonia and respiratory distress syndrome, which can cause death and varying degrees of sequelae.The pathogenesis of human adenovirus-induced severe adenovirus pneumonia remains unclear.It is currently believed that the main predisposing &factors for the occurrence and development of adenovirus pneumonia are immune dysfunction and immune dysfunction after viral infection.This article reviewed the research progress on the pathogenesis of adenovirus pneumonia induced by human adenovirus.
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OBJECTIVE： To evaluate the efficacy and safety of progesterone in the treatment of acute traumatic brain injury systematically， and to provide reference for clinical use. METHODS： Retrieved from Cochrane library， ClinicalTrials， Web of Science， PubMed， Embase， CBM， CNKI and Wanfang database， randomized controlled trial （RCTs） about progesterone （trial group） versus placebo or blank control （control group） in the treatment of acute traumatic brain injury were collected. After literature screening， data extraction and quality evaluation by risk bias evalution tool of Cochrane systematic evaluator manual 5.1.0， Meta-analysis was performed by using Rev Man 5.3 statistical software. RESULTS： A total of 10 RCTs were included， involving 2 652 patients. Results of Meta-analysis showed that there was no statistical significance in mortality[RR＝0.77，95％CI（0.56，1.07），P＝0.12]， the incidence of septicemia [RR＝1.11，95％CI（0.77，1.60），P＝0.59] or elevated liver enzymes[RR＝1.30，95％CI（0.68，2.50），P＝0.43]. The number of patients with favorable neurological outcome[RR＝1.23，95％CI（1.05，1.43），P＝0.008] in trial group was significantly more than control group. Results of subgroup analysis of mortality showed that there was no statistical significance in the mortality of patient’s GCS≤8 [RR＝0.79，95％CI（0.57，1.10），P＝0.16]， that of patient’s GCS≤12[RR＝0.69，95％CI（0.23，2.10），P＝0.52] or that of patient’s GCS ranging from 9 to 12 [RR＝0.78，95％CI（0.26，2.35），P＝0.65] between 2 groups. Results of subgroup analysis of neurological outcome showed that there was no statistical significance in the number of favorable neurological outcome of patient’s GCS≤8 [RR＝1.18， 95％CI（0.98，1.43），P＝0.09]， the number of favorable outcome of patient’s GCS≤12[RR＝1.15，95％CI（0.87，1.51），P＝0.32] and the number of favorable neurological outcome of patient’s GCS ranging from 9 to 12[RR＝2.07，95％CI（0.24，17.71），P＝0.51]. CONCLUSIONS： Progesterone can improve the prognosis of neurological function in patients with acute traumatic brain injury with good safety but cannot reduce mortality.
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Objective To evaluate the effect of intraperitoneally injected dexmedetomidine on abdominal adhesions in rats and the role of cholinergic anti-inflammatory pathway.Methods Forty cleangrade healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 4 groups (n =10 each) using a random number table method:sham operation group (Sham group),abdominal adhesion group (AA group),dexmedetomidine group (DEX group) and dexmedetomidine plus methyllycaconitine group (DEX-M group).The rat model of abdominal adhesions was established by cecal friction method.In Sham group,abdominal cavity was only opened and then sutured.Normal saline 2 ml was injected into the abdominal cavity and tail vein in group AA.In DEX and DEX-M groups,normal saline 2 ml and α7 nicotinic acetylcholine receptor antagonist methyllycaconitine 2.4 μg/g (dissolved in 2 ml normal saline) were injected,respectively,and dexmedetomidine 10μg/kg (dissolved in 2 ml normal saline) was intraperitoneally injected at the same time.The abdominal incision was opened under anesthesia at 7 days after establishing the model to observe the formation of abdominal adhesion,Phillips method was used for scoring,and enzyme-linked immunosorbent assay was used to determine the transforming growth factor-beta1 (TGF-β1) concentrations in ascites and tumor necrosis factor-alpha (TNF-α) concentrations in serum.The rats were then sacrificed,and the caecum tissue and its contralateral peritoneum and adhesion fibrous strips were obtained for examination of the pathological changes with a light microscope.Results Compared with group Sham,the abdominal adhesion score and serum TNF-α concentrations were significantly increased in AA and DEX-M groups,and the TGF-β1 concentration in ascites was significantly increased in AA,DEX and DEX-M groups (P＜0.05).Compared with group AA,the serum TNF-α concentrations and TGF-β1 concentration in ascites were significantly decreased in group DEX-M,and the abdominal adhesion score was significantly decreased (P＜0.05),and the pathological changes of caecum tissue,contralateral peritoneum and adhesion fibrous strips were significantly attenuated in group DEX.Compared with group DEX-M,the serum TNF-o concentrations were significantly increased (P＜0.05),no significant change was found in TGF-1 concentration in ascites (P＞0.05),and the pathological changes of caecum tissue,contralateral peritoneum and adhesion fibrous strips were accentuated in group DEX.Conclusion Intraperitoneally injected dexmedetomidine can mitigate abdominal adhesions,and the mechanism is related to activating cholinergic anti-inflammatory pathway and reducing systemic inflammatory response in rats.
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Objective@#To evaluate the effect of intraperitoneally injected dexmedetomidine on abdominal adhesions in rats and the role of cholinergic anti-inflammatory pathway.@*Methods@#Forty clean-grade healthy adult male Sprague-Dawley rats, weighing 220-250 g, were divided into 4 groups (n = 10 each) using a random number table method: sham operation group (Sham group), abdominal adhesion group (AA group), dexmedetomidine group (DEX group) and dexmedetomidine plus methyllycaconitine group (DEX-M group). The rat model of abdominal adhesions was established by cecal friction method.In Sham group, abdominal cavity was only opened and then sutured.Normal saline 2 ml was injected into the abdominal cavity and tail vein in group AA.In DEX and DEX-M groups, normal saline 2 ml and α7 nicotinic acetylcholine receptor antagonist methyllycaconitine 2.4 μg/g (dissolved in 2 ml normal saline) were injected, respectively, and dexmedetomidine 10 μg/kg (dissolved in 2 ml normal saline) was intraperitoneally injected at the same time.The abdominal incision was opened under anesthesia at 7 days after establishing the model to observe the formation of abdominal adhesion, Phillips method was used for scoring, and enzyme-linked immunosorbent assay was used to determine the transforming growth factor-beta1 (TGF-β1) concentrations in ascites and tumor necrosis factor-alpha (TNF-α) concentrations in serum.The rats were then sacrificed, and the caecum tissue and its contralateral peritoneum and adhesion fibrous strips were obtained for examination of the pathological changes with a light microscope.@*Results@#Compared with group Sham, the abdominal adhesion score and serum TNF-α concentrations were significantly increased in AA and DEX-M groups, and the TGF-β1 concentration in ascites was significantly increased in AA, DEX and DEX-M groups (P<0.05). Compared with group AA, the serum TNF-α concentrations and TGF-β1 concentration in ascites were significantly decreased in group DEX-M, and the abdominal adhesion score was significantly decreased (P<0.05), and the pathological changes of caecum tissue, contralateral peritoneum and adhesion fibrous strips were significantly attenuated in group DEX.Compared with group DEX-M, the serum TNF-α concentrations were significantly increased (P<0.05), no significant change was found in TGF-β1 concentration in ascites (P>0.05), and the pathological changes of caecum tissue, contralateral peritoneum and adhesion fibrous strips were accentuated in group DEX.@*Conclusion@#Intraperitoneally injected dexmedetomidine can mitigate abdominal adhesions, and the mechanism is related to activating cholinergic anti-inflammatory pathway and reducing systemic inflammatory response in rats.
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Objective@#To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.@*Methods@#A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using t test.@*Results@#According to the MTT assay results, EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments. Compared with the control group, the EGCG group showed significantly decreased number of Th1 cells (t = 3.43, P = 0.026) , increased number of Th2 cells (t = 6.68, P = 0.026) , and decreased Th1/Th2 ratio (P < 0.05) . The levels of IL-2 and IFN-γ in the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L, 1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ± 196.45 ng/L, 3 287.63 ± 235.54 ng/L respectively) , while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L, 285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L, 165.46 ± 32.25 ng/L respectively) . Compared with the control group, the EGCG group showed significantly decreased T-bet mRNA expression (t = 11.99, P < 0.001) , but increased GATA3 mRNA expression (t = 18.62, P < 0.001) .@*Conclusion@#EGCG can reduce the number of Th1 cells, inhibit the production of Th1 cytokines and transcription factors, and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors, followed by the modulation of Th1/Th2 immune imbalance.
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Objective To investigate the clinical pathologic features of a distinct variant of focal cortical dysplasia (FCD) characterized by neuronal loss of layer four.Methods Between 2005 and 2017,approximately 3 000 surgeries were performed for the treatment of intractable epilepsy at Xuanwu Hospital,Capital Medical University and Yuquan Hospital,Tsinghua University.Retrospective analysis of clinic-pathological data of patients with epilepsy surgery was made and histological manifestations of neuronal loss of cortical layer four were included in this study.Results In this cohort,25 patients (22 males and three females) were identified with early onset pharmaco-resistant epilepsy and regionally circumscribed neuronal loss of cortical layer four in surgical specimens from the occipital lobe.Histologically,except for neuronal loss in cortical layer four in all cases,glial scar lesions were found in some patients.Thus the histology of those cases can be subdivided into two groups:group A (13 cases):neuronal loss of cortical layer four without glial scar lesions;and group B (12 cases):neuronal loss of cortical layer four with glial scar lesions.Due to the prominent horizontal disorganization of cortical layering and lack of any other microscopically visible principle lesion,group A should be classified hitherto as FCD International League Against Epilepsy (ILAE) type Ⅰ b,however,group B with scar lesions and cortical dysplasia around the main leision,should be classified as FCD ILAE type Ⅲd.This retrospective analysis of clinical histories revealed a perinatal distress in 20 patients (80％),suggesting an acquired pathomechanism.Magnetic resonance imaging revealed abnormal signals in the occipital lobe in all patients,and signal changes suggestive of encephalomalacia were found in 18 patients.Surgical treatment achieved favorable seizure control (Engel class Ⅰ and Ⅱ) in 18 patients (75％ among 24 available follow up).Comparion of the two groups with age at epilepsy onset (group A:5.00±2.76,group B:5.01±3.78),the proportion of perinatal distress (group A:11/13,group B:9/12) and the follow-up results (favorable seizure control of the two groups was 9/13,9/11 respectively) showed that there was no statistically significant difference between the two groups.Conculsion Neuronal loss of cortical layer four in the occipital lobe should be classified as a distinct variant of FCD ILAE type Ⅲd.
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Objective To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.Methods A total of 33 patients with plaque-type psoriasis vulgaris were enrolled,and peripheral blood mononuclear cells (PBMC) were isolated and cultured.The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay.PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours.Flow cytometry was performed to determine proportions of Th 1 and Th2 cells,enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin [IL]-2,interferon [IFN]-γ) and Th2 cytokines (IL-4,IL-10) in the cell culture supernatant,and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor).Statistical analysis was carried out by using t test.Results According to the MTT assay results,EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments.Compared with the control group,the EGCG group showed significantly decreased number of Th1 cells (t =3.43,P =0.026),increased number of Th2 cells (t =6.68,P =0.026),and decreased Th1/Th2 ratio (P ＜ 0.05).The levels of IL-2 and IFN-γin the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L,1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ±196.45 ng/L,3 287.63 ± 235.54 ng/L respectively),while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L,285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L,165.46 ± 32.25 ng/L respectively).Compared with the control group,the EGCG group showed significantly decreased T-bet mRNA expression (t =11.99,P ＜ 0.001),but increased GATA3 mRNA expression (t =18.62,P ＜ 0.001).Conclusion EGCG can reduce the number of Th1 cells,inhibit the production of Th 1 cytokines and transcription factors,and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors,followed by the modulation of Th 1/Th2 immune imbalance.
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Objective To evaluate the effect of downregulation of microRNA (miR)-373 expression on cell cycle and apoptosis of a cutaneous squamous cell carcinoma (CSCC) cell line A431.Methods A431 cells at exponential growth phase were classified into 3 groups:miR-373 inhibitor group and negative control group transfected with miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.At 48 hours after the transfection,real-time PCR was performed to determine the expression of miR-373 in the above 3 groups,cell counting kit-8 (CCK-8) assay to evaluate the effect of downregulated expression of miR-373 on the proliferation of A431 cells,flow cytometry to investigate the distribution of cell cycle and changes in apoptosis of A431 cells in different treatment groups,and colorimetric analysis to detect the changes in caspase-3 activity in different treatment groups.Statistical analysis was carried out with SPSS 17.0 software by using two-sample t test for the comparison between two groups,one-way analysis of variance (ANOVA) for the comparison among 3 groups,and least significant difference (LSD)-t test for multiple comparisons.Results The expression of miR-373 was significantly lower in the miR-373 inhibitor group (0.120 ± 0.036) than in the untreated group (1.002 ± 0.022) and negative control group (1.037 ± 0.028,LSD-t =36.21,34.83,respectively,both P ＜ 0.001).At 48,72 and 96 hours,the miR-373 inhibitor group showed significantly decreased proliferative activity of A375 cells compared with the untreated group and negative control group (F =10.805,13.720 and 30.907 respectively,P =0.038,0.010 and 0.001 respectively).The proportion of A375 cells in G0/G1 phase was significantly higher in the miR-373 inhibitor group (64.69％ ± 1.18％) than in the untreated group (52.74％ ± 0.66％,t =15.51,P ＜ 0.001) and negative control group (53.80％ ± 0.80％,t =13.24,P ＜ 0.001).The proportion of total apoptotic cells and activity of caspase-3 in the miR-373 inhibitor group were 22.69％ ± 1.24％ and 1.238 ± 0.057 respectively,which were significantly higher than those in the untreated group (9.62％ ± 1.14％,0.413 ± 0.028 respectively,both P ＜ 0.001)and negative control group (9.66％ ± 0.97％,0.437 ± 0.036 respectively,both P ＜ 0.001).Conclusion MiR-373 may play an important role in the regulation of cell cycle and induction of apoptosis of the CSCC cell line A431.
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The theories of pathophysiology come from experimental research,and experimental teaching is an important part of pathophysiology course.Experimental teaching can cultivate the abilities of independent thinking and comprehensive analysis in students,improve their practical skills,and enhance their understanding and application of theoretical knowledge.However,teaching reform should be carried out due to the drawbacks of current pathophysiological experimental teaching.With the teaching idea centered on learning,the quality of pathophysiological experimental teaching can be enhanced by rational arrangement of experimental courses,optimization of teaching contents,and comprehensive application of various teaching models,so as to effectively improve the level of theoretical knowledge and comprehensive practical ability among students.
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The expression or dysfunction of long non-coding RNAs (lncRNAs) is closely related to various hereditary diseases, autoimmune diseases, metabolic diseases and tumors. LncRNAs were also recently recognized as functional regulators of fibrosis, which is a secondary process in many of these diseases and a primary pathology in fibrosis diseases. We review the latest findings on lncRNAs in fibrosis diseases of the liver, myocardium, kidney, lung and peritoneum. We also discuss the potential of disease-related lncRNAs as therapeutic targets for the clinical treatment of human fibrosis diseases.
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Humans , Autoimmune Diseases , Fibrosis , Genetic Diseases, Inborn , Kidney , Liver , Lung , Metabolic Diseases , Myocardium , Pathology , Peritoneum , RNA, Long NoncodingABSTRACT
Objective To investigate the expression of microRNA-373 (miR-373) in cutaneous squamous cell carcinoma (CSCC) tissues and cells,and to explore its effects on cell invasion.Methods Real-time PCR was performed to determine the expression of miR-373 in CSCC tissues and paralesional normal skin tissues,as well as in CSCC cell lines (A431 and SCL-1) and HaCaT cells.A431 cells were divided into 4 groups:miR-373 mimic group,miR-373 inhibitor group and negative control group which were transfected with miR-373 mimic,miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.Cell invasion assay was performed to evaluate effects of miR-373 downregulation on cell invasion.Western blot analysis was conducted to assess effects of miR-373 downregulation on the protein expression of matrix metalloproteinase-2 (MMP-2) and MMP-9.Results Expression of miR-373 was significantly higher in the CSCC tissues (2.465 ± 0.218) than in the paralesional normal skin tissues (1.000 ± 0.000,P ＜ 0.05),and higher in SCL-1 cells (1.864 ± 0.178) and A431 cells (2.919 ± 0.277) than in HaCaT cells (1.000 ± 0.000,P ＜ 0.05).Most notably,miR-373 expression was also markedly higher in metastatic CSCC tissues than in non-metastatic CSCC tissues (3.323 ± 0.344 vs.1.914 ± 0.161,t =4.158,P =0.000 4).Compared with the untreated group and negative control group,the miR-373 mimic group showed significantly increased miR-373 expression and invasive ability,while the miR-373 inhibitor group showed markedly decreased miR-373 expression and invasive ability (all P ＜ 0.05).Conclusion MiR-373 downregulation can significantly suppress the invasion of A431 cells,and obviously decrease the protein expression of MMP-2 and MMP-9.
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Objective To investigate the expression of microRNA-373 (miR-373) in cutaneous squamous cell carcinoma (CSCC) tissues and cells,and to explore its effects on cell invasion.Methods Real-time PCR was performed to determine the expression of miR-373 in CSCC tissues and paralesional normal skin tissues,as well as in CSCC cell lines (A431 and SCL-1) and HaCaT cells.A431 cells were divided into 4 groups:miR-373 mimic group,miR-373 inhibitor group and negative control group which were transfected with miR-373 mimic,miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.Cell invasion assay was performed to evaluate effects of miR-373 downregulation on cell invasion.Western blot analysis was conducted to assess effects of miR-373 downregulation on the protein expression of matrix metalloproteinase-2 (MMP-2) and MMP-9.Results Expression of miR-373 was significantly higher in the CSCC tissues (2.465 ± 0.218) than in the paralesional normal skin tissues (1.000 ± 0.000,P ＜ 0.05),and higher in SCL-1 cells (1.864 ± 0.178) and A431 cells (2.919 ± 0.277) than in HaCaT cells (1.000 ± 0.000,P ＜ 0.05).Most notably,miR-373 expression was also markedly higher in metastatic CSCC tissues than in non-metastatic CSCC tissues (3.323 ± 0.344 vs.1.914 ± 0.161,t =4.158,P =0.000 4).Compared with the untreated group and negative control group,the miR-373 mimic group showed significantly increased miR-373 expression and invasive ability,while the miR-373 inhibitor group showed markedly decreased miR-373 expression and invasive ability (all P ＜ 0.05).Conclusion MiR-373 downregulation can significantly suppress the invasion of A431 cells,and obviously decrease the protein expression of MMP-2 and MMP-9.
ABSTRACT
Objective Streptococcus sanguis is a possible candidate bacterium for the caries replacement therapy, which has no advantages in the acidic environment.The aim of the study was to construct acid-resistant strains of Streptococcus sanguis, determine its acid tolerance, and explore the mechanism of its antagonism against Sterptococcus mutans.Methods By gradually reducing the pH value of the medium, we constructed acid-resistant strains of Streptococcus sanguis, observed their growth and measured their acid tolerance according to their survival rate against lethal pH.We evaluated the competitive relationship between Streptococcus sanguis and Streptococcus mutans by plate experiment and detected the changes of related acid resistance genes by real-time quantitative PCR.Results The growth of Streptococcus sanguis and its acid-resistant strains were limited by the pH value, and that of Streptococcus sanguis was better in either acidic or normal environment.The lethal pH value of Streptococcus sanguis was 3.6, that of its acid-resistant strains was 2.3, and the survival rate of the acid-resistant strains was 66.59% in the pH 3.6 environment.In comparison, the lethal pH value of Streptococcus mutans was 2.5, that of its acid-resistant strains was 2.1, and the survival rate of the acid-resistant strains was 2.55% in the pH 2.5 environment.In the presence of chloramphenicol, the acid-resistant strains could not survive in the original lethal pH.In the sub-lethal pH environment, the expressions of the acid resistance-related genes Groel and Dnak in the acid-resistant strains were significantly up-regulated as compared with those in the original Streptococcus sanguis (P<0.05).Conclusion Streptococcus sanguis has an acid adaptability and can enhance acid resistance in the sub-lethal pH environment.Acid-resistant Streptococcus sanguis in the replacement therapy may provide some new ideas for the treatment of dental caries.