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Abstract Factor X deficiency ranks among the rarest coagulopathies and has a variable presentation spectrum. We intend to present a proposal for anesthesia protocol for individuals with the coagulopathy. The excision of an ovarian neoplasm was proposed for a 26-year-old, female, ASA II patient, with congenital Factor X deficiency. Physical examination and lab tests were normal, except for Prothrombin Time (PT) 22.1s (VR: 8-14s), International Normalized Ratio (INR) 1.99 (VR: 0.8-1.2) and Activated Partial Thromboplastin Time (aPTT) 41.4s (VR: 25-37s). We concluded that a history of bleeding should always be investigated, along with a pre-anesthetic coagulation study.
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Humans , Female , Adult , Blood Coagulation Disorders/diagnosis , Blood Coagulation Disorders/ethnology , Factor X Deficiency/complications , Anesthesia/adverse effects , Partial Thromboplastin Time , Prothrombin TimeABSTRACT
The MYC gene, one of the most common dysregulated driver genes in human cancers, is composed of three paralogous genes C-MYC, N-MYC and L-MYC. It is abnormally activated in more than half of cancer types. Since MYC plays an important role in the formation, maintenance and progression of cancer, targeting MYC is an effective strategy for cancer treatment. As a potential anti-cancer target, MYC is considered "undruggable" because it lacks a suitable pocket for accommodating small molecule inhibitors. Recently, under the guidance of protein structure information and many computational tools, many indirect strategies to inhibit MYC have emerged and shown favorable anti-cancer effects in tumor models. In this paper, the recent small molecules that indirectly target MYC are divided into inhibitors acting on the protein-protein interaction (PPI) among MYC and other proteins, and targeting inhibitors regulating MYC action. Additionally, the introduction and assessment towards compounds with different mechanisms are summarized to provide reference for the further research of MYC inhibitors.
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A retrospective analysis of 7 patients of multiple myeloma (MM) with initial manifestation of bleeding and coagulation abnormalities were performed. Clinical manifestations, laboratory and imaging examinations were collected. The activity of coagulation factors was measured before the treatment. Single factor X deficiency was seen in one patient. Two cases had factor Ⅶ deficiency, while four other patients had multiple factor deficiency. The time from onset of symptoms to diagnosis ranged from 2 to 10 months. After anti-MM treatment started and plasma or coagulation factors were transfused, the prolonged coagulation time returned to normal from 28-84 days. Most of these patients presented large, deep and multiple sites of hematoma, which caused concerns of bone marrow puncture and may direct to other differential diagnoses. This is helpful to improve physicians′ understanding of the special clinical characteristics in MM patients.
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The transforming growth factor-β (TGF-β) is a polypeptide cytokine, belonging to the TGF-β superfamily. TGF-β signaling pathway can regulate cell biological activity and play a bidirectional regulatory role in the development and progression of cancer. TGF-β signaling pathway can cause the occurrence and development of ovarian cancer through various processes including inducing cell excessive proliferation, influencing tumor microenvironment and regulating immune evasion. Targeted therapy with TGF-β as a biomar-ker for ovarian cancer provides a new way to improve the prognosis of patients.
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Objective@#To explore the effect of the umbilical cord mesenchymal stem cells(UC-MSCs) on the pulmonary fibrosis in silicosis rats.@*Methods@#SPF male Sprague Dawley rats were randomly divided into control group, silica model group and UC-MSCs treatment group with 12 rats each group. SiO2 intra-tracheal injection(0.5 ml of 50 mg/ml/rat) were applied to silica model group and UC-MSCs treatment groups. After that UC-MSCs treatment group received 1 ml UC-MSCs suspension (3×106 cells/ml) by tail vein injection on the 29th, 36th, 43th and 50th day after exposure to the first silica suspension. On the 60th and 75th day after exposure to silica suspension, all animals were examed for pulmonary CT. Then the rats were euthanized on 75th day after the first exposure to silica.Lung's histopathological examination of the rats from all the groups were carried out. The content of hydroxyproline in lungs, TGF-β1 and IL-6 in serum were examined.@*Results@#The lung's histopathological examination showed no obvious inflammatory cell and no fibrosis in the lung tissue of the control group, there were a lot of inflammatory cell aggregation and collagen fiber deposition in silica model group, while in the UC-MSCs intervention group and treatment group, there were less inflammatory cells and collagen fiber. The rats from silica model groups had higher HYP, TGF-β1 and IL-6 than the rats from UC-MSCs treatment group and control group. Lung fields of rats in the control group were clear and no obvious high-density shadow. Different-sized granular high-density shadows or reticular fibrous shadows were found diffusely distributed in the lungs of the rats in silica model group. Lung field of rats in UC-MSCs intervention group and treatment group were less high density shadows, and more clear.@*Conclusion@#UC-MSCs can alleviate the pulmonary fibrosis in silica model rats through regulating the secretion of some fibrosis related cytokines.
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Objective@#To investigate the effects and the potential mechanism of shikonin on rat random flaps.@*Methods@#Seventy-two wistar male rats in grade SPF were randomly divided into negative control group, tetramethylpyrazine group (TMP group) or shikonin treatment group. The random skin flap sized 8 cm×2 cm, with a cephalic based pedicle, was performed on the back of the rat. Each group was administered accordingly by intraperitoneal injection once per day for 7 days: shikonin treatment group (1 mg/kg), TMP group (10 mg/kg) and control group (1 ml/kg). Morphological changes of skin flaps were observed by HE staining. The positive expression of iNOS and COX-2 in skin flap tissues after operation were detected by immunofluorescence staining. The serum contents of TNF-α and IL-6 were detected by ELISA.@*Results@#Inflammatory cell infiltration and inflammatory reaction of flap was more severe in control group at different time points after operation. The number of inflammatory cells in shikonin treatment group and TMP group were significantly decreased, compared with controls (P<0.01). The decrease of the inflammatory cell numbers in shikonin treatment group was more significant. The proliferation of granulation tissue and fibroblast in skin flap was obvious, and the survival rates of the skin flap were significantly increased in shikonin treatment group and TMP group (both P<0.01). The numbers of iNOS or COX-2 positive cells in shikonin treatment group and TMP group were significantly decreased, when compared with controls (both P<0.01). Compared with TMP group, the numbers of iNOS and COX-2 positive cells in shikonin treatment group were significantly decreased (P<0.01 and P<0.05, respectively)in early period after operation. Compared with the control group, serum levels of TNF-α and IL-6 in shikonin treatment group and TMP group were significantly decreased (all P<0.01). The shikonin treatment group developed more significant reduce.@*Conclusions@#Shikonin can significantly improve inflammatory response of skin flap in rats, which might be related to inhibiting the expression of iNOS and COX-2, decreasing serum levels of TNF-α and IL-6 , and reducing the release of inflammatory cytokines .
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Objective@#To explore the relationship between protease-chymase secreted by mast cells, and activated transforming growth factor-β1(TGF-β1), in skin with secondary lymphedema(SLE)in lower extremity, so as toidentify the key factors in fibrosis of lymphedema.@*Methods@#In this study, the affected limb skin of 7 SLE patients was includedas the experimental group, and normal skin tissue of the lower limb of 7 volunteers was used as controls. The skin samples were assayed by Masson staining, and the expressions of chymase and TGF-β1 were assayed by immunohistochemistry, immunofluorescent staining and enzyme-linked immunosorbent assay.@*Results@#There was obvious fibrosis in the skin of lower extremity in patients with lymphedema. The number of MCs andthe expressions of chymase, latency-associated peptide TGF-β1 (LAP TGF-β1) and TGF-β1 were all significantly increased in fibrotic skin in lymphedema, compared with those in normal skin. At the same time, the chymase-containing mast cells accumulated in the lymphatic vessels, with higher expression of TGF-β1.@*Conclusions@#The expression of chymase and TGF-β1 was significantly increased in the fibrotic skin insecondary lower extremity lymphedema. The increased expression of chymase in the skin may activate more TGF-β1 expression, and the increased TGF-β1 may promote skin fibrosis in SLE.
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Objective@#To evaluate the clinical efficacy and safety of nimodipine combine with butylphthalide in the treatment of patients with mild to moderate vascular cognitive impairment(VCI).@*Methods@#From January 2012 to December 2016, 100 patients with mild to moderate VCI in Jinhua Municipal Central Hospital were randomly divided into control group(n=50) and treatment group(n=50) according to the random number table method.The control group received butylphthalide capsule, 200 mg po tid.The treatment group received nimodipine tablets, 40mg po tid, on the basis of the control group.The two groups of patients were treated for 24 weeks.Montreal cognitive assessment(MoCA), activities of daily living(ADL), serum hs-CRP, IL-6, TNF-α, clinical efficacy and adverse drug reactions were compared after treatment.@*Results@#After treatment, the scores of MoCA and ADL in the treatment group were (24.32±2.87)points, (59.22±6.17)points, respectively, which were significantly higher than those in the control group[(22.76±2.67)points, (55.63±6.3)points, t=2.814, 2.870, all P<0.05]. The effective rates in the treatment group and control group were 74.00%(37/50), 52.00%(26/50), respectively, and there was statistically significant difference between the two groups(χ2=5.191, P<0.05). After treatment, the levels of hs-CRP[(189.51±23.27)mg/L vs.(211.51±25.51)mg/L], IL-6[(76.42±9.86)ng/L vs.(95.85±10.23)ng/L], TNF-α[(0.24±0.08)ng/L vs.(0.32±0.10)ng/L] between the treatment group and the control group had statistically significant differences(t=4.505, 9.670, 4.417, all P<0.05). The adverse drug reactions were nausea and vomiting in 3 cases in the control group(6.00%), nausea and vomiting in 3 cases and hypotension in 1 case in the treatment group(8.00%), and there was no statistically significant difference between the two groups(P>0.05).@*Conclusion@#Nimodipine combined with butylphthalide in the treatment of mild to moderate VCI is effective and has high safety.
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Objective@#To investigate the effects of remifentanil combined with propofol on hemodynamics, inflammatory stress response and immune function in patients with acute abdomen complicated with septic shock.@*Methods@#From June 2017 to August 2018, 112 patients with acute abdomen complicated with septic shock who admitted to the First Affiliated Hospital of Xiamen University were enrolled in the study.They were randomly divided into observation group and control group according to the digital table, with 56 cases in each group.The control group was anesthetized with sevoflurane combined with propofol.The observation group was anesthetized with remifentanil combined with propofol.The hemodynamic parameters of the patients entering the operating room(T0), 0.5h(T1), 1h(T2) and awake(T3) after anesthesia were recorded.The intraoperative norepinephrine dosage was recorded.The inflammatory response, stress response and immune function indicators at T0, T2 and T3 were recorded.@*Results@#Compared with T0, T1 and T2, the MAP of the two groups was higher at T3, and the differences were statistically significant(tcontrol group=4.834, 4.484, 5.378, tobservation group=6.420, 7.006, 6.152, all P<0.05). Compared with T0, the HR of the two groups was higher at T3, and the differences were statistically significant(tcontrol group=5.943, tobservation group=7.722, all P<0.05). Compared with T1 and T2, CO of the two groups was higher at T3, and the differences were statistically significant(tcontrol group=4.276, 2.262, tobservation group=6.318, 5.132, all P<0.05). There were no statistically significant differences in MAP, HR and CO between the two groups at T0, T1, T2 and T3(all P>0.05). The doses of norepinephrine in the observation group and the control group were (1 587.7±287.5)μg and (1 937.9±397.6)μg, respectively, and the difference was statistically significant(t=5.341, P<0.05). The serum levels of C reactive protein(CRP), tumor necrosis factor alpha(TNF-α) and interleukin 6(IL-6) increased with time in the two groups, and the differences were statistically significant(tcontrol group=17.06, 36.13, 19.07, 3.822, 9.466, 2.874, 14.18, 26.87, 16.21, tobservation group=11.72, 20.79, 11.01, 2.810, 6.559, 3.716, 10.52, 24.56, 17.64, all P<0.05). At T2 and T3, the serum levels of CRP, TNF-α and IL-6 in the observation group[T2: CRP (89.63±17.65)mg/L, TNF-α (51.16±10.16)ng/L, IL-6 (34.26±6.25)ng/L, T3: CRP (136.15±26.25)mg/L, TNF-α (58.64±11.12)ng/L, IL-6 (67.56±12.67)ng/L]were lower than those in the control group[T2: CRP (112.15±22.34)mg/L, TNF-α (59.56±11.58)ng/L, IL-6 (42.65±8.37)ng/L, T3: CRP (175.16±34.75)mg/L, TNF-α (65.79±11.35)ng/L, IL-6 (79.02±14.56)ng/L], the differences were statistically significant(t=5.919, 6.703, 4.080, 3.367, 6.010, 4.443, all P<0.05). Compared with T0, serum levels of epinephrine(E), cortisol(Cor) increased in two groups at T2 and T3, and the differences were statistically significant(tcontrol group=10.03, 8.096, 8.679, 7.029, tobservation group=6.473, 4.728, 6.330, 4.727, all P<0.05). Compared with T2, serum levels of E and Cor in two groups at T3 were decreased, and the differences were statistically significant(tcontrol group=2.400, 2.638, tobservation group=2.260, 2.162, all P<0.05). At T2 and T3, the serum levels of E, Cor in the observation group[T2: E (286.36±41.02)ng/L, Cor (262.52±29.89)μg/L, T3: E (270.35±33.59)ng/L, Cor (253.23±30.28)μg/L]were lower than those in the control group[T2: E (312.56±38.75)ng/L, Cor (287.56±38.76)μg/L, T3: E (295.79±35.12)ng/L, Cor (270.25±30.15)μg/L], the differences were statistically significant (t=3.457, 3.917, 3.828, 2.981, all P<0.05). At T0, T2 and T3, there were no statistically significant differences in CD3+, CD4+, CD8+ and CD4+/CD8+ between the two groups(all P>0.05).@*Conclusion@#Remifentanil combined with propofol anesthesia can make hemodynamics more stable in patients with acute abdomen complicated with septic shock, and can alleviate inflammation and stress response.
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Objective@#To clarify the effect of TRAF2 in the biological behavior of gastric cancer and explore the mechanism.@*Methods@#TRAF2 stably depleted AGS cell was established. Cell growth was monitored by x-CELLigence system. Cell proliferation was detected using cell viability assay. The apoptosis and cell cycle were detected by flow cytometry. The difference of migration and invasion abilities were measured by real-time xCELLigence system and Transwell. The expression and activity of NF-κB signaling pathway were measured by western blot and TransAM assay. The expression of TRAF2 in gastric cancer tissue and its clinical significance were detected by immunohistochemistry.@*Results@#The cell index of AGS-siTRAF2 cells was significantly lower than that of AGS-sictrl cells at 8 h. In the cell viability assay, the A values of AGS-siTRAF2 cells were 51 296.00±2 631.06, 68 389.25±6 703.21 and 65 559.50±6 339.22 at 24 h, 48 h and 72 h. The values of the viability of AGS-siTRAF2 cells were significantly lower than those of AGS-sictrl cells (P<0.001). The results of flow cytometry showed that the apoptosis rates of AGS-siTRAF2 cells were (1.42±0.07)%, (2.98±0.11)% and (1.56±0.03)% at 24 h, 48 h and 72 h, respectively, which were significantly higher than those of AGS-sictrl cells (all P<0.05). The distribution of S phase in AGS-siTRAF2 cells was (23.57±1.12)%, while that in the AGS-sictrl cells was (19.49±1.19)%. The difference was statistically significant (P=0.012). AGS-siTRAF2 cells migrated much slower than AGS-sictrl cells from 3 h and the number of migrated AGS-sictrl cells was 121.7±6.7 while that of AGS-siTRAF2 cells was 84.0±6.6 (P=0.002). The cell index of AGS-siTRAF2 cells was less than that of AGS-sictrl cells from 3 h. In Transwell assay, the number of invaded AGS-sictrl cells was 109.3±3.1 after 24 h of culture, significantly higher than 79.0±6.2 of AGS-siTRAF2 cells (P=0.002). Western blot analysis showed that the expression levels of RelA, RelB, p50 and p52 in AGS-siTRAF2 cells were significantly lower than those in AGS-sictrl cells. The activities of RelA, RelB, p50 and p52 in AGS-siTRAF2 cells were 0.01±0.00, 0.01±0.01, 0.92±0.01 and 0.53±0.03, respectively, significantly lower than those of AGS-sictrl cells (all P<0.001). High TRAF2 expression (TRAF2-high) was found in 53.0% of GC samples, while TRAF2-high was only observed in 38.0% of the paired adjacent tissues (P=0.033). The expression of TRAF2 was significantly higher in the tubular adenocarcinoma, poor differentiation advanced T, advanced N, and clinical staging (P<0.05). The median survival time were 17 months and 78 months in the TRAF2 high-expression and low-expression groups, respectively, and the difference was statistically significant (P=0.010).@*Conclusions@#Depletion of TRAF2 inhibits the AGS cell growth, migration and invasion. The expression of TRAF2 is increased in gastric tumor tissue. The expression of TRAF2 is associated with the prognosis of gastric cancer.
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Objective@#To evaluate the protective effect of ulinastatin (UTI) on myocardial injury after post-cardiac arrest syndrome (PCAS) of cardiopulmonary resuscitation in pigs.@*Methods@#Twelve male 3-4 months pigs were randomly divided into two groups, UTI group and control group. The ventricular fibrillation (VF) animal model was replicated by programmed stimulation method. Among the 12 pigs, 11 pigs were successfully resuscitated by CPR after 5 min VF, of which, 6 pigs were in the UTI group and 5 pigs in the control group. Immediately after resuscitation, pigs in the UTI group was given 100 kU dissolved in 5 mL normal saline with slowly injection every 3 h until 24 h after recovery (no drug was given at 24 h). In the control group, 5 mL normal saline was given with same delivery time and frequency as that in the UTI group. The venous blood of the pigs was collected at VF, 2, 4, 6, 12, and 24 h after ROCS, and tumor necrosis factor -α (TNF-α), interleukin-6 (IL-6), superoxide dismutase (SOD), malondialdehyde (MDA), and ischemia modified albumin (IMA) were measured by enzyme-linked immunosorbent assay (ELISA) method. Because IMA was sensitive and increased rapidly, venous blood was detected at 1 h after ROSC, and the rest test time were same with the rest of the blood factors. Statistical analysis was performed using LSD-t test and variance analysis. The pigs were sacrificed 24 h after ROSC, and specimens from heart tissue were taken for HE staining.@*Results@#Before ventricular fibrillation in three groups, there was no significant difference in serum levels of inflammatory cytokines, oxidative stress indexes, and myocardial ischemia markers between the two groups (P>0.05). At 2 h after ventricular fibrillation, the levels of TNF-α and IL-6 level in the UTI group was significantly lower than those in the control group. At 4 h, MDA level in the UTI group was significantly lower than that in the control group (P<0.05); IMA was significantly increased at 1 h after ROSC, and the level of 1 h in the UTI group was significantly lower than that in the control group (P<0.05), but was not statistically significant at 12 h between the two groups (P>0.05). HE staining results showed that the damage degree of myocardial tissue in the UTI group was significantly lower than that in the control group at 24 h after ROSC.@*Conclusions@#UTI can significantly antagonize inflammatory response, reduce oxidative stress, and improve the myocardial tissue injury after resuscitation. It can protect myocardial injury after cardiopulmonary resuscitation.
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Objective@#To investigate the intervention effect of SB431542, which inhibits the TGF-β/Smad3 signaling pathway, on silicotic fibrosis in rats.@*Methods@#A total of 40 specific pathogen-free Sprague-Dawley rats were divided into normal saline control group, model group, SB431542 inhibitor group, and SB431542 inhibitor control group using a random number table, with 10 rats in each group. All rats except those in the normal saline control group were given non-exposed single intratracheal instillation of free silicon dioxide dust suspension 1 mL (50 mg/mL) ; the rats in the SB431542 inhibitor group were given intraperitoneal injection of SB431542 (5 mg/kg) on days 7 and 30 after dust exposure, those in the SB431542 inhibitor control group were given intraperitoneal injection of SB431542 cosolvent (5 mg/kg) on days 7 and 30 after dust exposure, and those in the normal saline control group were given intratracheal instillation of an equal volume of normal saline (5 mg/kg). On day 60 after dust exposure, the paraffin-embedded section of the right upper lobe of lung was collected for HE staining; the left upper lobe of lung was collected to measure the mRNA levels of fibronectin (FN) , collagen type I (COL-I) , and collagen type III (COL-III) by quantitative real-time PCR; the right inferior lobe of lung was collected to measure the protein levels of FN, COL-I, COL-III, phosphorylated Smad3 (p-Smad3) , and Smad3.@*Results@#Compared with the normal saline control group, the model group had nodules with various sizes in lung tissue, with rupture of some alveolar septa, emphysema changes, and pulmonary interstitial fibrosis, as well as significant increases in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . Compared with the SB431542 inhibitor control group, the SB431542 inhibitor group had a relatively complete structure of lung tissue without marked nodules and with a small amount of exudate in alveolar space and the lumen of bronchioles, as well as significant reductions in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 in lung tissue (P<0.05) . There were no significant differences in the mRNA expression of FN, COL-I, and COL-III and the protein expression of FN, COL-I, COL-III, p-Smad3, and Smad3 between the model group and the SB431542 inhibitor control group (P>0.05) .@*Conclusion@#SB431542 exerts an intervention effect on silicotic fibrosis by blocking the TGF-β/Smad3 signaling pathway and reducing the expression of the downstream fibrosis factors FN, COL-I, and COL-III.
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Klotho(KL)protein is a membrane-bound protein mainly expressed in the kidney and brain.It can regulate the expression of multiple signal pathways and target genes, such as ion channels, transforming growth factor(TGF), and exert anti-aging effects, kidney protection, anti-tumor and regulating metabolism.Recent studies have found that KL protein plays an important role in renal diseases such as renal interstitial fibrosis(RIF). KL deficiency can induce and promote the progress of RIF, and KL overexpression or supplementation can prevent RIF.This review focuses on the anti-RIF effect of KL protein through multiple signaling pathways, and aims to provide a new direction for anti-RIF therapy.
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Objective@#To investigate the effect of TNF-α on the expressions of tight junction protein ZO-1 and Claudin-4 in rat alveolar epithelial typeⅡ cells (AEC-Ⅱ).@*Methods@#Rat AEC-Ⅱ cells were cultured in vitro and divided into the control group, and TNF-α 24 h, 48 h, 72 h groups. The control group was cultured with DME F12 medium containing fetal bovine serum, and the TNF-α groups were intervened by TNF-α with a concentration of 10 ng/mL for 24 h, 48 h, and 72 h, respectively. After co-culture, the transmission electron microscopy was used to identify and observe the ultrastructural changes of rat AEC-Ⅱ cells. The cell inhibition rate was determined by MTT assay, and the cell apoptosis rate was measured by flow cytometry. The expression and distribution of tight junction protein ZO-1 and Claudin-4 in each group were observed by immunofluorescence method. The transcriptional levels of ZO-1 and Claudin-4 were detected by real-time fluorescent quantitative PCR. The expressions of ZO-1 and Claudin-4 were detected by Western blot. One-way analysis of variance (ANOVA) was used for comparisons among groups and SNK-q test was used for pairwise comparisons between groups. The non-parametric test of rank transformation was used when homogeneity of variance were not met. The value of P<0.05 was considered significantly different.@*Results@#Transmission electron microscopy showed that the AEC-Ⅱ cells in the control group obtained characteristic lamellar bodies of different sizes. The lamellar bodies in each TNF-α group were gradually emptied and apoptotic cells were observed under the visual field. The cell inhibition rate and early apoptosis rate of each TNF-α group were significantly higher than those of the control group (all P<0.05). Confocal microscopy showed that ZO-1 was linearly distributed along the cell membrane, and Claudin-4 was scattered along the cell membrane. In the TNF-α groups, the fluorescence intensity of ZO-1 was weakened and the continuity was broken as well as the linear structure. The fluorescence intensity and density of Claudin-4 were decreased in the TNF-α groups. Besides, the transcription and expression of ZO-1 in the TNF-α 24 h group were lower than those in the control group, but there was no significant difference (P>0.05). However, the mRNA level of ZO-1 in the TNF-α 48 h group (0.28±0.06) and 72 h group (0.13±0.07) were significantly lower than those in the control group. And their protein levels of TNF-α 48 h group (0.44±0.09) and of TNF-α 72 h group (0.2±0.01) were lower than that of the control group (0.69±0.12). Compared with the control group, the transcription level (24 h: 0.16±0.03; 48 h: 0.04±0.01; 72 h: 0.01±0.00 vs 1.00±0.00) and expression (24 h: 0.49±0.08; 48 h: 0.34±0.05; 72 h: 0.04±0.01 vs 0.96±0.13) of Claudin-4 in each TNF-α group were significantly decreased, and showed a decreasing trend with time (all P<0.05).@*Conclusions@#TNF-α can damage the pulmonary epithelial barrier by damaging alveolar epithelial typeⅡ cells and down-regulating the expression and distribution of ZO-1 and Claudin-4.
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Objective@#To analyze the changes of protein expression in aqueous humor of patients with primary open angle glaucoma (POAG), and to explore the biological markers and potential therapeutic targets associated with disease occurrence.@*Methods@#A retrospective case-control study was adopted.Ten patients with age-related cataract and 10 patients with POAG combined with cataract.were collected at the Tianjin Medical University Eye Hospital from October 2016 to December 2017.In the process of surgery, 100 μl of aqueous humor were collected with 1 ml syringe and No.1 needle through the surgical access.Those proteins extracted from aqueous humor were analyzed by quantitative proteomic mass spectrometry.The differential significance test was performed by Maxquant significances A. The differential proteins of the two groups were screened and determined with the conditions of P<0.05 and difference multiple>2.The biological big data was annotated by the function of GO and KEGG.The study was approved by the Ethics Committee of the Tianjin Medical University Eye Hospital (No.2013KY[L]-10). Patients and their guardians all signed the informed consent.@*Results@#Ninty-seven differential proteins were detected in this proteomic analysis, including 48 up-regulated proteins and 49 down-regulated proteins.GO analysis significantly differs in protein function involved in many aspects, some different proteins including lipopolysaccharide-binding protein (LBP), cluster of Differentiation 163(CD163), C-reactive protein (CRP), annexin A1 (ANXA1) were involved in inflammation; redox-related proteins were glutathione S-transferase P (GSTP1), thioredoxin (TXN), some different proteins related with cell adhesion movement were cartilage oligomeric matrix protein (COMP), desmocollin-2 (DSC2) and laminin subunit beta-2 (LAMB2); procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1), solid protein (tenascin, TNC) are associated with fibrosis; some different proteins related to nerve growth were reelin (RELN), semaphorin-3F(SEMA3F), semaphorin-4B(SEMA4B). Metabolism-related proteins included pyruvate kinase (PKM), carb oxypeptidase N subunit 2 (CPN2) and so on.KEGG analysis indicated that the expression of NrF2/ERK signaling pathway and TGF-β/NF-κB signaling pathway were different between the two groups.@*Conclusions@#The expression of GSTP1 and TXN in the aqueous humor of POAG is significantly decreased, which may regulate cell adhesion and activity and expression of extracellular matrix by regulating NrF2/ERK1/2 and TGF-β/NF-κB signaling pathways.GSTP1 and TXN may serve as a potential biomarker and therapeutic target of POAG.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) on behavioral changes, and expression of tyrosine hydroxylase (TH), α-synuclein(α-syn), transcription activating factor 6 (ATF6) and transcription factor X box binding protein 1 (XBP-1) in the substantia nigra of Parkinson's disease (PD) rats, so as to explore its mechanisms underlying improvement of motor function. METHODS: Thirty-six male SD rats were randomly divided into control, model and EA groups (n=12 rats in each group). The PD model was established by subcutaneous injection of rotenone (2 mg/kg) at the neck and back, once a day for 28 days. EA (2 Hz, 1 mA) was applied to "Fengfu" (GV16) and bilateral "Taichong" (LR3) for 20 min, once a day for 14 successive days. The voluntary motor behavioral changes (total distance, average speed, total movement time, total rest time in 8 min) were detected by open field tests. The immuno-activity of TH and α-syn in the substantia nigra was detected by immunohistochemistry, and the expression of ATF6 mRNA and XBP-1 mRNA detected by fluorescence real-time quantitative PCR. RESULTS: Following modeling and compared with the control group, the total distance, average speed and total movement time of voluntary movement were significantly decreased (P<0.01), and the total rest time was significantly increased (P<0.01). Moreover, the expression of TH was significantly decreased (P<0.01), and that of α-syn protein, ATF6 mRNA and XBP-1 mRNA significantly increased in the model group in comparison with the control group (P<0.01). After the intervention, the total distance, average speed, and total movement time of voluntary movement in the EA group were considerably higher than those in the model group (P<0.01), and the total rest time was obviously decreased in the EA group (P<0.01). The expression level of TH was significantly increased (P<0.01), and those of α-syn, ATF6 mRNA and XBP-1 mRNA were notably decreased in the EA group compared with the model group (P<0.01). CONCLUSION: EA intervention can improve the locomotor function in PD model rats, which is associated with its functions in up-regulating the expression of TH protein and down-regulating the expression of α-syn protein, and ATF6 mRNA and XBP-1 mRNA in the substantia nigra of mesencephalon.
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@# Objective: To validate the effect and the possible mechanism of human regulatory factor X1 (RFX1) over-expression on the proliferation and invasion of glioma F98 cells. Methods: RFX1-overexpressed F98 cells (F98-RFX1 group) were constructed by lentivirus transfection, a control group (F98-Vector group) and normal group (F98 group) were established. The effect of RFX1 over-expression on F98 cell proliferation was observed with counting method, cell apoptosis was determined by AnnexinV-PI staining, and the cell invasion was observed with Transwell method. Results: F98 cell line over-expressing RFX1 was successfully established. The proliferation capacity of F98-RFX1 group was significantly lower than that of F98 group (48 h: [12.08±2.17]×104/ml vs [23.67±4.51]×104/ ml, P<0.05] and F98-Vector group (96 h: [8.17±0.31]×104/ml vs [18.58±1.18]×104/ml, P<0.05); The apoptosis level of cells in F98RFX1 group was significantly increased ([21.89±2.33]% vs [3.38±1.39]%, [10.42±1.83]%, P<0.05]; The invasiveness of cells in F98RFX1 group was significantly reduced ([33.3±7.99] vs [56.5±13.9], [60.6±11.8], P<0.01). Conclusion: RFX1 can regulate the expression of genes related with proliferation and invasion, thereby inhibiting the proliferation and invasion ability of glioma cells and promote cell apoptosis.
ABSTRACT
Liver fibrosis is a common pathological response in chronic liver injury. In the pathological process of hepatic injury, signaling pathways associated with hepatic fibrosis, which mediates the repair, proliferation and fibrosis of the liver secrete different cytokines. In these pathways, transforming growth factor beta (TGFβ) and signal transducer and activator of transcription 3 (STAT3) play key roles in the proliferation and activation of hepatic stellate cells (HSCs) and promote epithelial mesenchymal transition. In addition, it is also involved in the process of proliferation and transformation of collagen and extracellular matrix molecules into myofibroblasts. TGFβ and STAT3 molecular-related signaling pathways mediate the loss of epithelial phenotype and gene expression in mature epithelial cells, transforming them into mesenchymal cells, and producing anti-apoptosis to hepatocytes and promoting the proliferation of HSCs. However, the mechanisms by which STAT3 and TGFβ molecules are involved in the development and progression of liver fibrosis are not sound distinct. In this review, we attempt to know the mechanisms and interactions of TGFβ and STAT3 molecules that mediate potential liver fibrosis, and promote their role in promoting HSCs production and epithelial mesenchymal transition.
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Objective@#To investigate the effect of transforming growth factor(TGF)-β1 stimulation on the proliferation of lung fibroblasts and the pathogenesis of bronchopulmonary dysplasia(BPD).@*Methods@#The lung fibroblasts of newborn rats were isolated, purified and identified on the 1st day after birth.The primary lung fibroblasts were stimulated by TGF-β1, and the protein expression of cyclin-dependent kinase-2(CDK2) and p27 were detected by immunohistochemistry and Western blotting.Real-time fluorescence quantitative polymerase chain reaction(PCR) was used to detect the mRNA expression of CDK2 and p27.CCK8 kit was used to detect cell proliferation, and cell cycle was detected by flow cytometry.@*Results@#The expression of CDK2 protein and mRNA in primary fibroblasts was increased after TGF-β1 stimulation in lung fibroblasts of newborn rats, accompanied by the decrease of p27 protein and mRNA.CCK8 showed that the value of A increased significantly, proliferation significantly increased the proportion of S phase cells at the same time, G0/G1 phase decreased.The percentage of S phase cells after stimulation with 0 μg/L, 10 μg/L TGF-β1 was 4.63%, 67.09%, respectively.The percentage of G0/G1 phase was 83.67% and 67.09%, respectively.The percentage of S phase cells was 7.64% and 9.11% respectively after stimulated with 10 μg/L TGF-β1 for 12 h and 48 h, respectively.The percentage of G0/G1 phase was 73.99% and 59.81% respectively.With the increase of TGF-β1 levels (0 μg/L, 5 μg/L and 10 μg/L), CDK2 protein and mRNA expression further increased; and the same level of TGF-β1 stimulation, stimulated for 12, 24 and 48 h, the expression of CDK2 protein and mRNA increased further, and increased significantly at 48 h (P<0.05 and <0.01 respectively). The levels of p27 protein and mRNA decreased further(all P<0.01).@*Conclusions@#TGF-β1 can up-regulate the expression of CDK2/p27 in primary lung fibroblasts of neonatal rats, accelerating cell cycle progression and over-proliferation of fibroblasts.
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Objective@#To investigate the effect and regulatory mechanism of Smurf2 on the negative regulator Smad7 of TGF-β1/Smad signaling pathway in hypertrophic scar fibroblasts.@*Methods@#From January to October 2014, 8 patients with hypertrophic scar after burn were admitted. The age of patients ranged from 1 year 8 months to 7 years, and the time of scar hyperplasia ranged from 2 to 12 months. The residual normal skin of the same patient was used as the control. The fibroblasts were isolated from hypertrophic scar and normal skin respectively and cultured. The third to fifth passage cells were used for the experiments. ① The protein expression of Smad7 in the two groups was detected by Western Blot. ② Hypertrophic scar fibroblasts and normal skin fibroblasts were treated with exogenous TGF-β1 at concentration of 10 ng/ml for 0 min, 5 min, 15 min, 30 min, 1 h, 2 h and 12 h. The expression of Smad7 protein and mRNA were detected by Western Blot and RT-PCR, respectively. ③ The cell lysates of the two groups were collected and incubated with the ubiquitin mixture for 1 h, 2 h and 6 h at 37℃, respectively. The degradation level of Smad7 protein was detected by Western Blot. ④ The cell lysate of hypertrophic scar fibroblasts was collected and incubated with ubiquitin mixture with or without proteasome inhibitor (MG132: MG115=1: 1) to study its inhibitory effect on the degradation of Smad7 in vitro. ⑤ Immunoprecipitation (IP) technique was used to detect the interaction between Smad 7 and E3 ubiquitin ligase Smurf2 in hypertrophic scar fibroblasts. ⑥ The expression of Smad7 protein in hypertrophic scar fibroblasts stimulated by TGF-β1 after Smurf2 silencing by small interference RNA (siRNA) technique were detected by Western blot.@*Results@#① There was no significant difference in the expression level of Smad7 protein between hypertrophic scar and normal skin fibroblasts(t=0.76, P=0.46). ② Expressions of Smad7 mRNA and protein in normal skin fibroblasts stimulated by exogenous TGF-β1 gradually increased in a time-dependent manner(P<0.05). The expression of Smad7 mRNA in the hypertrophic scar fibroblasts increased at all-time points except at 5min , (P<0.05), while there was no significant difference in the expression level of Smad7 protein at all-time points with or without TGF-β1 stimulation(P>0.05). ③ Degradation of Smad7 protein was enhanced in hypertrophic scar fibroblasts (the expression level of Smad 7 protein at each time point was compared with that of the control group and the last time point, P<0.05), while there was no significant difference in Smad7 protein degradation in normal skin fibroblasts(P=0.162). ④ Enhanced degradation of Smad7 in hypertrophic scar fibroblasts was blocked by the addition of the proteasome inhibitors MG132/MG115. ⑤ In hypertrophic scar fibroblasts, the Smurf2-Smad7 complex was detected, which indicated the interaction between Smurf2 and Smad7 in hypertrophic scar fibroblasts. ⑥ The expression of Smad7 protein was not increased in the hypertrophic scar fibroblasts stimulated by TGF-β1, whereas the stimulation of TGF-β1 increased the expression of Smad7 protein after silencing of Smurf2 gene expression.@*Conclusions@#In the hypertrophic scar fibroblasts, Smurf 2 attenuates the inhibitory effect of Smad 7 on TGF-β1 signaling pathway through the degradation of Smad7 by ubiquitination, which may be involved in the formation of hypertrophic scar.