;Aim To study the effect of exopolysaccharide from Trichoderma pseudokoningii (EPS) on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods The proliferation of HCT116 cells treated with EPS was examined by CCK-8 assay. The effect of EPS on the clone formation of HCT116 cells was detected by crystal violet staining. Cell apoptotic rate was determined by flow cytometry. The changes of mitochondrial membrane potential were observed by JC-1. The expressions of apoptosis-related proteins Bcl-2 and Bax were analyzed by Western blot, and caspase-9, caspase-8 and caspase-3 expressions were detected by the kit. Results EPS dose- and time-dependently inhibited the proliferation of the HCT116 cells in the range of 0 -800 mg • L " 1 . With increase of EPS concentration, the colony-formation ability of HCT116 cells decreased; the proportion of apoptotic cells increased and mitochondrial membrane potential decreased; the expression of Bcl-2 protein decreased, while the expression of Bax protein increased; the ratio of Bcl-2/Bax decreased gradually; caspase-9, caspase-8 and caspase-3 activities significantly increased. Conclusions EPS can inhibit proliferation of HCT116 and induce its apoptosis by up-regulating expression of Bax, caspase-9,caspase-8, caspase-3 and down-regulating the expression of Bcl-2.

AIM: To study protective effects of the extract from Toona sinensis seeds on gastric mucosal injury in experimental mice. METHODS: The mice were given 200 mg/kg, 400 mg/kg of Toona sinensis seeds every day for 2 weeks and ethanol or aspirin was used to induce gastric mucosa injury model. The gastric mucosal injury index and injury inhibition rate were calculated, the levels of SOD, MDA in serum and the contents of ET-1, PGE

Objective To investigate the expression, function and significance of long non-coding RNA (lncRNA) CASC19 in colorectal cancer (CRC). Methods Real-time quantitative polymerase chain reaction was employed to determine the expression of CASC19 in 40 paired samples from CRC surgical specimens and 5 CRC cell lines. The correlations of CASC19 expression with clinicopathologic parameters were analyzed. Transwell assay was applied to detect the migration ability of CRC cells after the CASC19 expression was knocked down by small interfering RNA. Results The expression of CASC19 in colorectal cancer was significantly higher than those in adjacent normal mucosa tissues (t=5.527, P<0.000 1) and was associated with metastasis (P=0.044). Knockdown of CASC19 expression in CRC inhibited the migration ability of CRC in vitro. Conclusions The expression of CASC19 increases in CRC. CASC19 expression is not associated with age, gender, or tumor site/differentiation but with tumor size, lymph node metastasis, and distant metastasis, suggesting high CASC19 expression may promote CRC metastasis.

AIM: To study the polymorphism distribution of methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR) genes and their influence on serum homocysteine (Hcy) concentration. METHODS: A total of 148 patients diagnosed with ischemic stroke from November 2020 to February 2021 in Yijishan Hospital of Wanan Medical College were selected for the study, and patients were typed for MTHFR 677C/T and MTRR 66A/G genes using fluorescent staining in situ hybridization technique. Serum Hcy concentrations were measured in 21 patients using a circulating enzyme assay. The distribution of MTHFR 677C/T and MTRR 66A/G gene polymorphisms were analyzed, and the differences in serum Hcy concentrations between patients with different genotypes were compared. RESULTS: The mutation rates of MTHFR 677C/T and MTRR 66A/G genes were 42.57% and 26.01%, respectively, and no significant differences in gene distribution frequencies were observed between men and women (P>0.05). The mean Hcy serum concentration was (16.04±4.34) μmol/L in 21 patients, including 8 patients (38.10%) with <15 μmol/L and 13 patients (61.90%) with ≥15 μmol/L. The Hcy serum concentrations in patients with different genotypes of MTHFR were TT (18.91±5.34) μmol/L, CT (14.38±1.84) μmol/L and CC (13.58±2.86) μmol/L, respectively, and were statistically different (P<0.001). Serum Hcy concentrations in patients with different genotypes of MTRR were not statistically different (P>0.05). CONCLUSION: MTHFR gene polymorphisms can affect serum Hcy concentrations. The MTHFR genotyping can be considered for individualized folic acid supplement. This conclusion should be further verified by expanding the clinical sample size.

Rationale: Trichosporon, an anamorphic fungus, proliferates under high humidity, causing serious opportunistic infections collectively called trichosporonosis. Among the Trichosporon species causing trichosporonosis are Trichosporon (T.) asahii, T. asteroides, T. cutaneum etc. Patient concerns: A 38-year-old Chinese male with severe aplastic anemia was admitted due to multiple joints pain, poor appetite, and right ankle swelling. One year earlier he had undergone allogeneic hematopoietic stem cell transplantation. Diagnosis: T. asahii infection and severe aplastic anemia. Interventions: Combined treatment of amphotericin B liposomes (55 mg/24 h) and voriconazole (200 mg/12 h) for 8 days. Outcomes: The symptoms of the patient's ankle were relieved and effusion cultures showed no T. asahii. Lessons: To the best of our knowledge, T. asahii ankle cavity effusion infections are rare. Trichosporon infections may be attributed to risk factors such as improper long-term use of antimicrobials for an underlying disease (e.g., anemia, hypoalbuminemia). Attention should be paid to prevent and control Trichosporon infections in order to avoid comorbidities.

OBJECTIVE: To explore the effect of the composition ratio on substitution of sulfate group in sulfated exopolysaccharide (EPS) from and how sulfate modification affects the anti-tumor activity of EPS. METHODS: We used a chlorosulfonic acid-pyridine method to modify EPS and analyzed the effect of esterification ratio on the degree of sulfate substitution using barium chloride turbidimetry. The sulfate groups binding with EPS were analyzed with infrared spectrum analysis. CCK-8 assay was used to evaluate the inhibitory effect of EPS sulfate (SEPS) on the proliferation of human colon cancer HCT 116 cells, and annexin V-FITC/PI double staining was used to assess the pro-apoptotic effect of SEPS in the cells. RESULTS: The esterifying agent and EPS at the composition ratios of 1:1 and 2:1 resulted in sulfate substitution of 0.98% (SEPS-1) and 1.18% (SEPS-2), respectively, and the substitution was improved by increasing the ratio of the esterifying agent ( < 0.05). Infrared spectrum analysis showed that the S=O stretching vibration absorption peak of -OSO appeared near 1249 cm, indicating that the sulfate group combined with EPS to form sulfate. CCK-8 assay showed that SEPS-1 produced stronger inhibitory effects on the proliferation of HCT 116 cells than EPS within the concentration range of 0.02-0.10 mg/L ( < 0.05). At the concentrations of 0.04-0.08 mg/L, SEPS-2 showed a lower anti-tumor activity than SEPS-1 ( < 0.05). SEPS-1 also showed stronger pro-apoptotic effect than EPS, and as its concentration increased, SEPS-1 dose-dependently increased the ratio of early apoptotic cells and necrotic cells; the cells treated with 0.06, 0.08 and 0.10 mg/mL SEPS-1 showed early apoptotic rates of 6.38%, 11.8% and 12.5%, and late apoptotic and necrotic rates of 5.26%, 8.04% and 6.80%, respectively. CONCLUSIONS The composition ratio of the esterifying agent has a direct impact on the degree of substitution of EPS, which can be improved by increasing the ratio of the esterifying agent. Sulfate modification of EPS can enhance its antitumor activity, which, however, is not directly related with the degree of substitution.

Objective: To investigate the values of T lymphocyte-bound complement activation products such as T-C3d and T-C4d, B lymphocyte-bound complement activation products such as B-C3d and B-C4d and erythrocyte-bound complement activation products such as E-C3d and E-C4d in the diagnosis of systemic lupus erythematosus (SLE). Methods: Peripheral blood samples from 68 SLE patients, 70 patients with non-SLE autoimmune diseases and 68 healthy controls were collected randomly, and the expression levels of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in these samples were detected by flow cytometry. Meanwhile, antinuclear antibodies (ANA), anti-double stranded DNA antibodies (anti-dsDNA), peripheral blood cell count and other markers were also detected. The differences of cell-bound complement activation products in three groups were analyzed with the area under the receiver operating characteristic curve (AUC), nonparametric test, sensitivity and specificity. Results: The specific median fluorescence intensity (SMFI) of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d in SLE patients were significantly higher than those in the patients with non-SLE autoimmune diseases and healthy controls (all P＜0.05). The SMFI (median \[P 25, P 75\]) of T-C4d, B-C4d and E-C4d in SLE patients were 3.8(1.2, 13.1), 22.1(6.2, 67.9) and 19.6(1.8, 52.4), respectively. The SMFI of T-C4d, B-C4d and E-C4d in SLE patients with reduced red blood cells and/or lymphocytes were significantly higher than that with normal red blood cell and lymphocyte count (all P＜0.05). The AUCs of T-C4d, B-C4d, E-C4d, T-C3d, B-C3d and E-C3d were 0.711, 0.763, 0.663, 0.631, 0.611 and 0.615, respectively (all P＜0.05). The sensitivity of the combination of T-C4d with B-C4d (73.5%) in the diagnosis of SLE was superior to that of anti-dsDNA (36.8%). Conclusion The cell-bound complement activation products (CB-CAPs) are specifically expressed in SLE patients, and their combination detection is helpful for the diagnosis of SLE.

OBJECTIVE: To investigate the spectrum-effect relationship between the total anthraquinone extract of Cassia seeds and fluorouracil (5-Fu)-induced liver injury in mice and identify the effective components in the extract. METHODS: A mouse model of liver injury was established by intraperitoneal injection of 5-Fu, with bifendate as the positive control. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and myeloperoxidase (MPO), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in the liver tissue were detected to investigate the effect of the total anthraquinone extract of Cassia seeds (0.4, 0.8 and 1.6 g/kg) on liver injury induced by 5-Fu. HPLC fingerprints of 10 batches of the total anthraquinone extracts were established to analyze the spectrum- effectiveness of the extract against 5- Fu- induced liver injury in mice and screen the effective components using the grey correlation method. RESULTS: The 5- Fu- treated mice showed significant differences in liver function parameters from the normal control mice (P < 0.05), suggesting successful modelling. Compared with those in the model group, serum ALT and AST activities were decreased, SOD and T- AOC activities significantly increased, and MPO level was significantly lowered in the mice treated with the total anthraquinone extract (all P < 0.05). HPLC fingerprints of the 31 components in the total anthraquinone extract of Cassia seeds showed good correlations with the potency index of 5-Fu-induced liver injury but with varying correlation strengths. The top 15 components with known correlations included aurantio-obtusina (peak 6), rhein (peak 11), emodin (peak 22), chrysophanol (peak 29) and physcion (peak 30). CONCLUSION The effective components in the total anthraquinone extract of Cassia seeds, including aurantio-obtusina, rhein, emodin, chrysophanol, and physcion, are coordinated to produce protective effects against 5-Fu-induced liver injury in mice.
Animals ; Mice ; Emodin ; Cassia ; Chemical and Drug Induced Liver Injury, Chronic ; Anthraquinones ; Antioxidants ; Fluorouracil/adverse effects* ; Plant Extracts/pharmacology*

This study aimed to assess whether chrysin(ChR) can inhibit epithelial-mesenchymal transition(EMT) of type Ⅱ alveolar epithelial cell and produce anti-pulmonary fibrosis effect by regulating the NF-κB/Twist 1 signaling pathway. Sixty rats were randomly divided into the control group, the bleomycin(BLC) group, BLC+ChR(50 mg·kg~(-1)) group and BLC+ChR(100 mg·kg~(-1)) group, with 15 rats in each group. The pulmonary fibrosis model was induced by intratracheal injection of BLC(7 500 U·kg~(-1)). Rats were orally administered with different doses of ChR after BLC injection for 28 days. The cells were divided into control group, TGF-β1 group(5 ng·mL~(-1)), and TGF-β1+ChR(1, 10, 100 μmol·L~(-1)) groups. The type Ⅱ alveolar epithelial cells were treated with TGF-β1 for 24 h, and then treated with TGF-β1 for 48 h in the presence or absence of different doses of ChR(1, 10 and 100 μmol·L~(-1)). The morphological changes and collagen deposition in lung tissues were analyzed by HE staining, Masson staining and immunohistochemistry. The mRNA and protein expression levels of collagen Ⅰ, E-cadherin, zonula occludens-1(ZO-1), vimentin, alpha smooth muscle actin(α-SMA), inhibitor of nuclear factor kappa B alpha(IκBα), nuclear factor-kappa B p65(NF-κB p65), phospho-NF-κB p65(p-p65) and Twist 1 in lung tissues and cells were detected by qPCR and Western blot, respectively. The animal experiment results showed that as compared with the BLC group, after administration of ChR for 28 days, bleomycin-induced pulmonary fibrosis in rats was significantly relieved, collagen Ⅰ expression in lung tissues was significantly reduced(P<0.05 or P<0.01), and EMT of alveolar epithelial cells was obviously inhibited [the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], concomitantly with significantly reduced IκBα and p65 phosphorylation level in cytoplasm and decreased NF-κB p65 and Twist 1 expression in nucleus(P<0.05 or P<0.01). The cell experiment results showed that different doses of ChR(1, 10 and 100 μmol·L~(-1)) significantly reduced TGF-β1-induced collagen Ⅰ expression(P<0.05 or P<0.01), significantly inhibited EMT of type Ⅱ alveolar epithelial cells[the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], and inhibited IκBα and p65 phosphorylation in cytoplasm and down-regulated NF-κB p65 and Twist 1 expression in nucleus induced by TGF-β1(P<0.05 or P<0.01). The results suggest that ChR can reverse EMT of type Ⅱ alveolar epithelial cell and alleviate pulmonary fibrosis in rats, and its mechanism may be associated with reducing IκBα phosphorylation and inhibiting NF-κB p65 phosphorylation and nuclear transfer, thus down-regulating Twist 1 expression.
Alveolar Epithelial Cells/metabolism* ; Animals ; Epithelial-Mesenchymal Transition ; Flavonoids ; NF-kappa B/metabolism* ; Rats ; Signal Transduction ; Transforming Growth Factor beta1/genetics*

To investigate the effects of salt-inducible kinase 2 (SIK2) on energy metabolism in rats with cerebral ischemia-reperfusion. Adult SD male rats were divided into 5 groups: sham group, ischemia group, reperfusion group, adenovirus no-load group, and SIK2 overexpression group with 5 animals in each group. The middle cerebral artery occlusion (MCAO) was induced with the modified Zea-Longa line thrombus method to establish the cerebral ischemia reperfusion model. Eight days before the MCAO, SIK2 overexpression was induced by injecting 7 μL adenovirus in the right ventricle, then MCAO was performed for followed by reperfusion HE staining was used to observe the pathological changes of cerebral tissue in rats; TTC staining was used to observe the volume of cerebral infarct. The levels of adenosine triphosphate (ATP) and adenosine diphosphate (ADP) in rat brain tissue were detected by ELISA; the levels of SIK2 and hypoxia-inducible factor 1α (HIF-1α) in the rat brain tissues were detected by RT-qPCR and Western blotting. Compared with the sham group, SIK2 level was decreased in the ischemia group, and it was further declined in the reperfusion group (<0.05). Compared with the sham group and ischemic group, the pathological injury in reperfusion group were more severe, and the infarct size was larger; compared with the reperfusion group and adenovirus no-load group, the pathological injury of the SIK2 overexpression group was milder, and the infarct size is less. Compared with the sharn group, HIF-1α was increased in both ischemia group and reperfusion group, especially in ischemia group (all <0.05); HIF-1α level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05). ATP level in ischemia group and reperfusion group was lower than that in the sham group, and the reperfusion group decreased more significantly than the ischemia group (<0.05); ADP content was increased in the ischemia and reperfusion group, and the ADP content in reperfusion group was significantly higher than that in the ischemia group (<0.05). ATP level in the SIK2 overexpression group was higher than that in the reperfusion group and adenovirus no-load group (all <0.05), and ADP was decreased in the SIK2 overexpression group (all <0.05). SIK2 can up-regulate the ATP level and down-regulate the ADP level in rat brain tissue and alleviate cerebral ischemia-reperfusion injury by increase the level of HIF-1α.
Animals ; Brain Ischemia ; Energy Metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics* ; Infarction, Middle Cerebral Artery ; Male ; Protein-Serine-Threonine Kinases ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury