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ObjectiveTo explore the inhibitory effect of water extract of Broussonetiae Fructus on hepatocellular carcinoma （HCC） induced by diethyl nitrosamine （DEN） in mice based on homologous phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B （PTEN/PI3K/Akt） signaling pathway. MethodThe primary HCC mouse model was constructed by intraperitoneal injection of DEN solution， and the HCC mice were randomly divided into model group， sorafenib group （0.01 g·kg-1·d-1）， low-dose Broussonetiae Fructus water extract group （0.9 g·kg-1·d-1）， medium-dose Broussonetiae Fructus water extract group （1.8 g·kg-1·d-1）， and high-dose Broussonetiae Fructus water extract group （3.6 g·kg-1·d-1）， with 10 mice in each group. Another 10 C57BL/6 mice were selected as a control group and intraperitoneally injected with an equal volume of normal saline. Mice were treated with different concentrations of Broussonetiae Fructus water extract when liver cancer-like white nodules appeared. sorafenib group was treated with sorafenib. The control group and model group were intraperitoneally injected with normal saline. The activities of alkaline phosphatase （ALP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， and γ-glutamyl transferase （γ-GT） in the serum of mice were detected by the biochemical analyzer. The expression levels of alpha-fetoprotein （AFP） and carcinoembryonic antigen （CEA） were detected by enzyme-linked immunosorbent assay （ELISA）. The degree of hepatocyte canceration and hepatocyte injury were observed by Hematoxylin-eosin （HE） and Masson staining. The proliferation of HCC cells was observed by immunohistochemical staining. The apoptosis of HCC cells in mice was observed by erminal-deoxynucleotidyl transferase mediated nick end labelling （TUNEL） staining. The expression levels of PTEN， PI3K， Akt， and p-Akt proteins related to the PTEN/PI3K/Akt signaling pathway were detected by Western blot. ResultCompared with the control group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the model group were significantly increased （P<0.01）. Carcinogenesis and inflammatory cell infiltration were obvious in liver tissue of mice， and a large number of blue collagen fiber hyperplasia was found. The number of Ki67 positive cells was significantly increased （P<0.01）， and the expression level of PTEN protein was significantly decreased， while PI3K and p-Akt protein expression was increased （P<0.01）. Compared with the model group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the medium-dose and high-dose Broussonetiae Fructus water extract groups were significantly decreased （P<0.05， P<0.01）. The degree of carcinogenesis and inflammatory cell infiltration in liver tissue were reduced， and the collagen fiber hyperplasia was significantly reduced. The number of Ki67 positive cells was significantly decreased， and the number of TUNEL positive apoptotic cells was significantly increased （P<0.05， P<0.01）. PTEN protein expression was increased， while p-Akt protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionThe water extract of Broussonetiae Fructus has a significant inhibitory effect on DEN-induced primary HCC in mice， and its mechanism may be related to the regulation of key protein expressions in the PTEN/PI3K/Akt signaling pathway.
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ObjectiveAt present, there are few reports on the effect of tRF 31 on breast cancer. This study aims to detect and analyze the expression level of tRF 31 in breast cancer tissues and to explore its effect on the proliferation of breast cancer cells.Methods32 tumor tissue samples and the corresponding para-cancer tissues from breast cancer patients in The Affiliated Cancer Hospital of Nanjing Medical University from November 2017 to February 2019 were collected. Six out of the thirty two samples were selected for high-throughput sequencing and at last tRF 31 (FC=6.5781, P=0.023) was selected as the research object. The expression of tRF-31 in cancer and para-cancer tissues was measured by RT-PCR. The sensitivity and specificity of tRF-31 expression in the breast cancer diagnosis were analyzed by ROC curve. Two kinds of human breast cancer cell lines (MDA-MB-231 and MCF-7) were divided into two groups: control group (transfected with negative control) and tRF-31 group (transfected with tRF-31 inhibitor). The proliferation of transfected breast cancer cells was detected by CCK-8 and clonal formation assay. The expression changes of threonine kinase (AKT) and mammalian target of rapamycin (mTOR) in breast cancer cells after transfection were measured to explore the relationship between TRF-31 and AKT/mTOR signaling pathway.ResultsThe expression of tRF-31 in cancer tissues (0.103±0.207) was significantly higher than that in para-cancerous ones (0.028±0.039). ROC curve showed that the sensitivity and specificity of tRF-31 in detecting breast cancer were 90.63% and 53.13%, respectively. In MDA MB 231 cells, the expression of tRF-31 in the tRF-31 group was significantly lower than that in the control group [(0.267±0.012) vs (1±0.040), P<0.01)], and in MCF-7 cells, the expression of tRF-31 in the tRF-31 group was also significantly lower than that in the control group. In MDA-MB-231 and MCF-7 cells, the proliferation ability of the tRF-31 group was lower than that of the control group at 0h, 24h, 48h and 72h. In MDA-MB-231 cells, the clonal formation rate of tRF-31 group [(43.67±3.29)%] was significantly lower than that of the control group [(100±3.74)%] (P<0.01). In MCF-7 cells, the clonal formation rate of tRF-31 group [(49±2.94)%] was significantly lower than that of the control group [(100±4.89)%] (P<0.01). In MDA-MB-231 and MCF-7 cells, the protein levels of AKT and mTOR in tRF-31 group were significantly lower than those in the control group.ConclusiontRF-31 is highly expressed in breast cancer tissues and can promote the proliferation of breast cancer cells. This result is expected to provide a new target for the treatment of breast cancer.
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Objective: To comprehensively evaluate the different passivation materials on growth and cadmium uptake and accumulation of Ophiopogon japonicus cultivated in cadmium contaminated soil of. Methods: Using one-year-old O. japonicus as experimental material, the effects of five different passivation materials (white marble Ar, straw biochar Br, fly ash Fh, bacterial residue Me, diatomite Dm) on the growth, physiological and biochemical indexes and cadmium absorption and accumulation of O. japonicus were studied by soil pot experiment. Results: The biomass, chlorophyll, soluble sugar and soluble protein of O. japonicus could be significantly increased in soils contaminated with two kinds of Cd under different passivation materials. Among them, Ar and Me were the most improved. In addition, the activities of antioxidant enzyme system of O. japonicus were significantly decreased, and the content of MDA had no noteworthy change. Different passivation materials remarkably increased the content of flavonoids in the underground part of O. japonicus, among which Ar and Fh treatments increased the most. The contents of heavy metal Cd in all parts of O. japonicus were decreased by different passivation materials, and the contents of Cd in aboveground and underground parts of O. japonicus were significantly lower than those of the control, and the effects of Ar, Br and Fh treatments were better than those of the control. Conclusion: Studies have shown that by combining and comparing treatments of different passivating materials, in terms of the biomass of O. japonicus treated with Me, the biomass improvement effect of Me was better than that of Ar and Br. Aiming at the effect of reducing Cd absorption and accumulation of O. japonicus and improving the effective component flavone of O. japonicus, it can be concluded that Ar and Fh have better effects in treating Cd polluted soil and improving the quality of O. japonicus.
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Objective@#To compare and analyze patient’s general condition, laboratory testing and therapeutic responses of isolated immunoglobulin G4- related sclerosing cholangitis (IgG4-SC) and immunoglobulin G4 sclerosing cholangitis combined autoimmune pancreatitis (IgG4-SC/AIP).@*Methods@#A retrospective study was conducted on IgG4-SC patients who attended outpatient and inpatients department of our hospital from April 2014 to March 2018 and their demographic characteristics, laboratory testing, and therapeutic responses were statistically analyzed. Normal distribution of continuous variables was compared with t-test, non-normal distribution of continuous variables was compared using the Mann-Whitney U test, and the categorical variables were compared with χ 2 test.@*Results@#29 IgG4-SC patients were included, including 19-isolated IgG4-SC and 10 IgG4-SC combined AIP (IgG4-SC/AIP). The average age of onset in the isolated IgG4-SC group was (46.06±19.03) years which was lower than IgG4-SC/AIP group (62.60±15.11), t = -2.360, P < 0.05. The median IgG4 in IgG4-SC/AIP patients is higher than that in isolated IgG4-SC, respectively 10.87 (3.73 ~ 20.13) and 3.14 (2.37 ~ 4.78)g/L(U = 159.000, P < 0.05). IgG4/IgG ratio is higher in IgG4-SC/AIP, than that in isolated IgG4-SC, respectively 0.62(0.23 ~ 0.86) and 0.16(0.10 ~ 0.21), U = 130.000, P < 0.05. Liver cirrhosis was more common in isolated IgG4-SC group (47%) than the IgG4-SC/AIP group (0), χ 2 = 9.637, P < 0.05. The median biochemical response time of isolated IgG4-SC group was 3.00 (2.00 to 4.00) months, which was longer than 1.00 (1.00 to 1.25) months of IgG4-SC/AIP group, U = 30.000, P < 0.05. The biochemical recurrence rate of isolated IgG4-SC group was 32%, which was lower than that of IgG4-SC/AIP (χ 2 = 6.461, P < 0.05).@*Conclusion@#Serum IgG4 level and IgG4/IgG ratio were higher in patients with IgG4-SC/AIP group, and therapeutic responses in isolated IgG4-SC patients were worse than that of IgG4-SC/AIP patients. The efficacy of glucocorticoid monotherapy and immunosuppressive agents combined with glucocorticoid therapy demonstrated no considerable difference in IgG4-SC patients.
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Objective Few studies are reported on the correlation of Spinster homolog2(Spns2) with cancer.This study aims to investigate the expression of Spns2 in gastric cancer and the adjacent tissue and explore its effects on the proliferation, clone formation and migration of human gastric cancer cell line SGC-7901.Methods Samples of gastric cancer and the adjacent tissue were collected from 20 patients (12 males and 8 females) after surgery between February 2016 and August 2016. The expression of Spns2 in the gastric cancer and adjacent tissues was determined by immunohistochemistry. Eukaryotic expression vector pEX-1 (pGCMV/MCS/EGFP/Neo)-Spns2 and siRNA were constructed and transfected into the human gastric cancer cell line SGC-7901. The cells were divided into a pEX-1- Spns2 (siRNA-Spns2) group and a negative control pEX-1 (siRNA-NC) group. The mRNA and protein expressions of Spns2 were detected by RT-PCR and Western blot, respectively. And the effects of Spns2 on the migration, clone formation and proliferation of the SGC-7901 cells after Spns2 over-expressed and down-regulated were evaluated by cell counting kit-8 (CCK-8) assay, clone formation assay, transwell experiment and wound healing scratch assay.Results The expression of Spns2 was significantly lower in the gastric cancer than in the adjacent tissue (0.39±0.04 vs 0.45±0.02, P<0.05). After transfection, both Spns2 mRNA and protein levels were remarkably higher in the pEX-1-Spns2 (21.96±2.08 and 10.71±2.78) than in the pEX-1 group (0.88±0.12 and 0.83±0.16) (P<0.05), but markedly lower in the siRNA-Spns2 (0.35±0.07 and 0.53±0.07) than in the siRNA-NC group (0.91±0.09 and 0.92±0.08) (P<0.05). At 48, 72 and 96 hours, the proliferation of the SGC-7901 cells was significantly decreased in the pEX-1-Spns2 group as compared with the pEX-1 group (P<0.05) but increased in the siRNA-Spns2 group in comparison with the siRNA-NC group (P<0.05); the clone formation rate was reduced in the pEX-1-Spns2 group as compared with the pEX-1 group (\[33.30±3.81\]％ vs \[48.00±4.60\]％, P<0.05) but elevated in the siRNA-Spns2 group in comparison with the siRNA-NC group (\[48.38±4.22\]％ vs \[26.25±4.88\]％, P<0.05).Conclusion Spns2 is lowly expressed in gastric cancer, and its over-expression in SGC-7901 cells can reduce its proliferation, clone formation and migration. Spns2 may play a role in inhibiting the development and progression of gastric cancer.
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Autoimmune hepatitis (AIH)is a chronic progressive immune-mediated inflammatory liver disease. Both adults and children could be involved and women are more often affected. AIH is characterized by elevation of serum IgG/hypergammaglobulinemia,autoantibodies and interface hepatitis with diverse clinical spectrum. The incidence of AIH is increasing in recent years. If not treated timely,it may lead to cirrhosis and liver failure. Immunosuppressive therapy is crucial for improving patients'survival rate. This article summarized the advances in diagnosis and treatment of AIH.
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OBJECTIVE@#To investigate the distribution and contents of vimentin (Vim) and glial fibrillary acidic protein (GFAP) immunoreactivities in the central nervous system (CNS) of normal newborn, adult and aged rats.@*METHODS@#In this study, thirty healthy and normal Sprague-Dawley rats were simply classified into three groups: Newborn (7 days aged), adult (5 months aged) and aged (24 months aged) rats. Brains and spinal cord were dissected and cut into frozen sections. The expression of Vim and GFAP in CNS were detected by confocal immunofluorescence.@*RESULTS@#In each group, Vim was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was highest in neuron-like cell of newborn rats, while Vim was mainly expressed in cell bodies in adult and aged rats. GFAP was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was in astrocytes of aged rats. In the third ventricle, Vim was detected in all groups, and only observed in neuron-like cells of newborn. Meanwhile, the GFAP expression showed no significant differences between adult and aged rats in this region. The co-localization of Vim and GFAP were mainly observed in hippocampus and cerebral cortex of newborn, but this co-localization was found in the third ventricle of the rats in all groups.@*CONCLUSION@#Our data demonstrate for the first time that the expression of Vim and GFAP in the rat's CNS during development. This data may provide a foundation for the further mechanistic studies of these two main intermediate filaments during development of CNS.
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Objective To investigate the distribution and contents of vimentin (Vim) and glial fibrillary acidic protein (GFAP) immunoreactivities in the central nervous system (CNS) of normal newborn, adult and aged rats. Methods In this study, thirty healthy and normal Sprague–Dawley rats were simply classified into three groups: Newborn (7 days aged), adult (5 months aged) and aged (24 months aged) rats. Brains and spinal cord were dissected and cut into frozen sections. The expression of Vim and GFAP in CNS were detected by confocal immunofluorescence. Results In each group, Vim was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was highest in neuron-like cell of newborn rats, while Vim was mainly expressed in cell bodies in adult and aged rats. GFAP was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was in astrocytes of aged rats. In the third ventricle, Vim was detected in all groups, and only observed in neuron-like cells of newborn. Meanwhile, the GFAP expression showed no significant differences between adult and aged rats in this region. The co-localization of Vim and GFAP were mainly observed in hippocampus and cerebral cortex of newborn, but this co-localization was found in the third ventricle of the rats in all groups. Conclusion Our data demonstrate for the first time that the expression of Vim and GFAP in the rat's CNS during development. This data may provide a foundation for the further mechanistic studies of these two main intermediate filaments during development of CNS.