Lactic Acid Inhibits the Cytotoxicity of NK-92 Cells via Activating the Lactate Receptor GPR81

Aims: Lactate acid functions as not only an energy source but a signaling molecule through the lactate receptor GPR81 under physiological conditions. However, the pathological role of lactic acid in the tumor microenvironment remains unclear, particularly for immune cells. Methodology: NK-92 cells were treated with L-lactic acid solutions at final concentrations of 10, 20, 30, and 40 mM, and its cell viability and cytotoxicity on A549 cells and A375 cells were evaluated by CCK8 assay and crystal violet assay, respectively. Furthermore, qPCR was used to assess the expression of GPR81 and cytotoxicity-related genes in NK-92 cells treated with antagonist and agonist. And their relationship between lactate/GPR81 pathway and cytotoxicity-related genes were analyzed by Pearson’s correlation. Results: The viability of NK-92 cells was inhibited by L-lactic acid with increasing concentration. Additionally, the cytotoxic activity against tumor cells of NK-92 cells treated with L-lactic acid decreased with increasing concentration. Moreover, qPCR results demonstrated that GPR81 can be activated by lactic acid or agonist (3,5-DHBA) and downregulate the expression cytotoxicity-related genes which included FASLG gene(Fas Ligand),TNF-? gene(Tumor necrosis factor-?), INFG gene (Interferon-?), RPF1 gene (Perforin 1), GZMA gene (Granzyme A), GZMB gene (Granzyme B), GZMH gene (Granzyme H), GAMK gene (Granzyme K) and GZMM gene (Granzyme M). And the expression of GPR81 returned to near-control level when treated with L-lactic acid in the presence of antagonist (3-OBA), the expression of cytotoxicity-related genes did as well. Pearson’s correlation analysis of cytotoxicity-related genes with GPR81 revealed that their correlation coefficient seems negative. Conclusion: Lactic acid can activate the GPR81 to downregulate the expression of cytotoxicity-related genes, subsequently lower the cytotoxicity of NK-92 cells.

Construction of a prognostic model for hepatocellular carcinoma based on pyroptosis-related genes / 中华肝脏病杂志

Objective: To study the construction of a prognostic model for hepatocellular carcinoma (HCC) based on pyroptosis-related genes (PRGs). Methods: HCC patient datasets were obtained from the Cancer Genome Atlas (TCGA) database, and a prognostic model was constructed by applying univariate Cox and least absolute shrinkages and selection operator (LASSO) regression analysis. According to the median risk score, HCC patients in the TCGA dataset were divided into high-risk and low-risk groups. Kaplan-Meier survival analysis, receiver operating characteristic (ROC) curves, univariate and multivariate Cox analysis, and nomograms were used to evaluate the predictive ability of the prognostic models. Functional enrichment analysis and immune infiltration analysis were performed on differentially expressed genes between the two groups. Finally, two HCC datasets (GSE76427 and GSE54236) from the Gene Expression Omnibus database were used to externally validate the prognostic value of the model. Univariate and multivariate Cox regression analysis or Wilcoxon tests were performed on the data. Results: A total of 366 HCC patients were included after screening the HCC patient dataset obtained from the TCGA database. A prognostic model related to HCC was established using univariate Cox regression analysis, LASSO regression analysis, and seven genes (CASP8, GPX4, GSDME, NLRC4, NLRP6, NOD2, and SCAF11). 366 cases were evenly divided into high-risk and low-risk groups based on the median risk score. Kaplan-Meier survival analysis showed that there were statistically significant differences in the survival time between patients in the high-risk and low-risk groups in the TCGA, GSE76427, and GSE54236 datasets (median overall survival time was 1 149 d vs. 2 131 d, 4.8 years vs. 6.3 years, and 20 months vs. 28 months, with P = 0.000 8, 0.034 0, and 0.0018, respectively). ROC curves showed good survival predictive value in both the TCGA dataset and two externally validated datasets. The areas under the ROC curves of 1, 2, and 3 years were 0.719, 0.65, and 0.657, respectively. Multivariate Cox regression analysis showed that the risk score of the prognostic model was an independent predictor of overall survival time in HCC patients. The risk model score accurately predicted the survival probability of HCC patients according to the established nomogram. Functional enrichment analysis and immune infiltration analysis showed that the immune status of the high-risk group was significantly decreased. Conclusion: The prognostic model constructed in this study based on seven PRGs accurately predicts the prognosis of HCC patients.

Risk modeling based on HER-2 related genes for bladder cancer survival prognosis assessment / 北京大学学报(医学版)

OBJECTIVE@#To investigate the correlation between the human epidermal growth factor receptor-2-related genes (HRGs) and survival prognosis of bladder cancer and to construct a predictive model for survival prognosis of bladder cancer patients based on HRGs.@*METHODS@#HRGs in bladder cancer were found by downloading bladder tumor tissue mRNA sequencing data and clinical data from the cancer genome atlas (TCGA), downloading HER-2 related genes from the molecular signatures database (MsigDB), and crossing the two databases. Further identifying HRGs associated with bladder cancer survival (P < 0.05) by using single and multi-factor Cox regression analysis and constructing HRGs risk score model (HRSM), the bladder cancer patients were categorized into high-risk and low-risk groups accor-ding to the median risk score. Survival analysis of the patients in high- and low-risk groups was conducted using R language and correlation of HRGs with clinical characteristics. A multi-factor Cox regression analysis was used to verify the independent factors affecting the prognosis of the patients with bladder cancer. The area under the curve (AUC) of the receiver operating characteristic curve (ROC) of HRSM was calculated, and a nomogram was constructed for survival prediction of the bladder cancer patients. Analysis of HRSM and patient immune cell infiltration correlation was made using the TIMER database.@*RESULTS@#A total of 13 HRGs associated with patient survival were identified in this study. Five genes (BTC, CDC37, EGF, PTPRR and EREG) were selected for HRSM by multi-factor Cox regression analysis. The 5-year survival rate of the bladder cancer patients in the high-risk group was significantly lower than that of the patients in the low-risk group. High expression of PTPRR was found to be significantly and negatively correlated with tumor grade and stage by clinical correlation analysis, while EREG was found to be the opposite; Increased expression of EGF was associated with high grade, however, the high expression ofCDC37showed the opposite result. And no significant correlation was found between BTC expression and clinical features. Correlation analysis of HRSM with immune cells revealed a positive correlation between risk score and infiltration of dendritic cells, CD8+T cells, CD4+T cells, neutrophils and macrophages.@*CONCLUSION@#HRGs have an important role in the prognosis of bladder cancer patients and may serve as new predictive biomarkers and potential targets for treatment.

Changes of hippocampal gray matter volume and immune related genes in dizocilpine-induced schizophrenia model rats / 中华行为医学与脑科学杂志

Objective:To investigate the changes of hippocampal gray matter volume and expression of candidate immune related genes in a rat model of schizophrenia established by repeated administration of dizocilpine(MK-801).Methods:Thirty SPF grade Sprague-Dawley male rats at postnatal day 28 were randomly divided into MK-801 medium-dose (0.25 mg/kg) group, MK-801 high-dose(0.50 mg/kg) group and normal saline (5 mL/kg) group according to random number table method, with 10 in each group.Rats were given continuous intraperitoneal administration according to grouping once a day for 14 days.Open field test, novel object recognition test and Y-maze test were used at postnatal day 60 to detect spontaneous activity, exploration ability, anxiety level, object recognition memory ability and spatial working memory of rats, respectively.At postnatal day 67, structural magnetic resonance imaging was used to detect the changes of hippocampal gray matter volume in rat.And at postnatal day 70, qRT-PCR was used to detect the expression of candidate immune-related genes in rat hippocampus.SPSS 25.0 was used for statistical analysis, one-way ANOVA was used for comparison among multiple groups, and Tukey test was used for further pairwise comparisons.Results:(1)The behavioral results showed that there were significant differences in the total movement distance, central area activity time, novel object recognition index, and spontaneous correct alternation rate among the three groups ( F=11.15, 10.11, 13.62, 11.99, all P<0.05). The total movement distances in MK-801 medium-dose group and MK-801 high-dose group ((21.44±2.17) m, (22.87±1.96)m) were higher than that in the normal saline group ((18.70±1.88) m) (both P<0.05). The activity time of the central area in the MK-801 medium-dose group and MK-801 high-dose group((3.24±1.58) s, (2.50±1.32) s) were lower than that of the normal saline group ((6.05±2.48)s) (both P<0.01). Novel object recognition indexes in the MK-801 medium-dose group and MK-801 high-dose group((56.10±3.99)%, (54.00±6.41)%) were both lower than that in the normal saline group ((65.90±5.65)%)(both P<0.01), and the rates of spontaneous correct alternation ((54.60±7.03)%, (51.60±8.84)%) in the two groups were lower than that of the normal saline group ((68.40±8.57)%) (both P<0.01). (2) The results of structural magnetic resonance imaging showed that there were significant differences in the volume of hippocampal gray matter among the three groups ( F=9.24, P<0.001). The volumes of hippocampal gray matter in MK-801 medium-dose group and MK-801 high-dose group were lower than that in normal saline group(both P<0.001). (3)By constructing protein-protein interaction network, four candidate immune related genes were screened out: neuropeptide Y (NPY), somatostatin (SST), cholecystokinin (CCK) and tachykinin 1 (TAC1). The results showed that the mRNA expression levels of NPY, SST and CCK in the hippocampus of the three groups were significantly different ( F=11.41, 10.43, 5.85, all P<0.05), but there was no statistical difference in the TAC1 mRNA expression level ( F=0.08, P>0.05). The mRNA levels of NPY, SST and CCK in the hippocampus of rats in the MK-801 high-dose group were lower than those in the normal saline group (all P<0.05). Conclusion:Both medium dose and high dose MK-801 administration can reduce the volume of hippocampal gray matter in schizophrenia model rats, but they have different effects on the expression of hippocampal immune related genes, of which high dose administration has a greater effect.

Quantification of phytochemicals in Muli bamboo [Melocanna baccifera (Roxb.) Kurz] leaf and shoot extracts and immunomodulatory effect of dietary ethanolic leaves extract on Labeo rohita (Hamilton)

Traditionally, bamboo has immense medicinal values owing to their rich bioactive compounds. On the other hand, scientific developments in global aquaculture have revealed the significance of dietary nutraceuticals in aquaculture. In this study, efforts have been made to evaluate the properties of shoot and leaf extracts of Melocanna baccifera (Roxb.) Kurz (Muli bamboo) and demonstrated its immunomodulatory potential. At first, four different extracts of bamboo leaf and shoot were prepared through water and ethanolic solvents followed by phytochemical profiling, antioxidant and antimicrobial activities of extracts. Based on the in vitro evaluation, BLAL (bamboo leaf alcoholic) extract was chosen for further in vivo evaluation. Second experiment was carried to assess the toxicity of BLAL extract on the Indian major carp, Labeo rohita (Hamilton), locally called ‘rohu’. No mortality was observed up to 20 g of extract kg-1 body weight. Additionally, haemolytic assay was also conducted to ascertain the cellular toxicity of extract. Third experiment was conducted to find out the effect of dietary BLAL extract [doses: control (0.0%), T1 (0.01%), T2 (0.1%) and T3 (1%) of extract kg-1 feed] on immune related genes (HSP70, IL-1? and TNF-?) expression in L. rohita. The present study confirms the presence of vital phytochemicals in bamboo extracts and immunomodulatory potential of ethanolic leaf extract in roh

Prognosis prediction model for hepatocellular carcinoma based on autophagy related genes / 生物医学工程学杂志

Autophagy is a programmed cell degradation process that is involved in a variety of physiological and pathological processes including malignant tumors. Abnormal induction of autophagy plays a key role in the development of hepatocellular carcinoma (HCC). We established a prognosis prediction model for hepatocellular carcinoma based on autophagy related genes. Two hundred and four differentially expressed autophagy related genes and basic information and clinical characteristics of 377 registered hepatocellular carcinoma patients were retrieved from the cancer genome atlas database. Cox risk regression analysis was used to identify autophagy-related genes associated with survival, and a prognostic model was constructed based on this. A total of 64 differentially expressed autophagy related genes were identified in hepatocellular carcinoma patients. Five risk factors related to the prognosis of hepatocellular carcinoma patients were determined by univariate and multivariate Cox regression analysis, including TMEM74, BIRC5, SQSTM1, CAPN10 and HSPB8. Age, gender, tumor grade and stage, and risk score were included as variables in multivariate Cox regression analysis. The results showed that risk score was an independent prognostic risk factor for patients with hepatocellular carcinoma ( HR = 1.475, 95% CI = 1.280-1.699, P < 0.001). In addition, the area under the curve of the prognostic risk model was 0.739, indicating that the model had a high accuracy in predicting the prognosis of hepatocellular carcinoma. The results suggest that the new prognostic risk model for hepatocellular carcinoma, established by combining the molecular characteristics and clinical parameters of patients, can effectively predict the prognosis of patients.

A novel metastasis-related genes-based signature for predicting progression-free interval of papillary thyroid carcinoma / 中华内分泌外科杂志

Objective:We aimed to build a novel model with metastasis-related genes (MTGs) signature for predicting progression-free interval (PFI) of papillary thyroid carcinoma (PTC) .Methods:We integrated PTC datasets with the MTGs to identify differentially expressed MTGs (DE-MTGs), then we established a novel MTGs based signature and validated it in external datasets and cell lines. Finally, we established a signature and clinical parameters-based nomogram for predicting the PFI of PTC.Results:We identified 155 DE-MTGs related to PFI in PTC. The functional enrichment analysis showed that the DE-MTGs were associated with essential oncogenic processes. Consequently, we established and optimized a novel 10-gene signature. The novel signature had a C-index of 0.76 and the relevant nomogram had a C-index of 0.80. Also, it was closely related to pivotal clinical characters of multiple datasets and invasiveness of PTC cell lines. And the signature was an independent prognostic factor in PTC. Finally, we built a nomogram including the signature and relevant clinical factors. The efficacy was satisfying in predicting PTC’s PFI.Conclusions:The MTG signature and nomogram were closely associated with PTC prognosis and may help clinicians improve the individualized prediction of PFI.

Effect of Jiangtang Xiaozhi Tablet on Liver Circadian Clock-related Genes CLOCK， BMAL1， REV-ERBα， and REV-ERBβ in Mice with Metabolic Dysfunction-associated Fatty Liver Disease / 中国实验方剂学杂志

ObjectiveTo explore the effect of Jiangtang Xiaozhi tablet （JTXZT） on metabolic dysfunction-associated fatty liver disease and to study the mechanism from the perspective of circadian clock-related genes such as circadian locomotor output cycles kaput （CLOCK）， brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 （BMAL1）， reverse-eritroblastosis receptor （REV-ERB）α and β. MethodA total of 50 male SPF C57BL/6J mice were randomized into normal group （n=10） and modeling group （n=40）. The normal group was fed with normal diet， and the modeling group with high-fat diet for 4 weeks. Then the model mice were randomly classified into model group， high-dose （12.5 g·kg-1） and low-dose （6.25 g·kg-1） Jiangtang Xiaozhi tablet groups， and orlistat group （70 mg·kg-1）， with 10 mice in each group. The normal group and model group received equivalent volume of distilled water （8 weeks）. Then， the body weight of mice was measured， and the content of serum triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） was determined with biochemical method. Serum content of free fatty acid （FFA） and leptin was detected by enzyme-linked immunosorbent assay （ELISA）. Pathological changes of liver tissue and epididymal adipose tissue were observed based on hematoxylin-eosin （HE） staining. Liver fibrosis was examined based on Masson's trichrome staining， and changes of lipids based on oil red O staining. The expression of CLOCK， BMAL1， REV-ERBα， and REV-ERBβ was detected by Western blot and immunohistochemistry assay. ResultCompared with the normal group， the model group had high content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin （P<0.05， P<0.01）， showed ballooning degeneration and focal microvesicular steatosis of liver cells， enlarged adipocytes， and inflammatory cell clusters and fibrous tissue hyperplasia， and displayed increased protein expression of sterol regulatory element binding protein （SREBP） 1 and peroxisome proliferators-activated receptor （PPAR）γ （P<0.01） and decreased protein expression of PPARα （P<0.05）， CLOCK， BMAL1， REV-ERBα and β （P<0.05， P<0.01）. Compared with the model group， JTXZT-H group down-regulated the content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin in mice （P<0.05， P<0.01）， and the JTXZT groups demonstrated reduction in the degree and range of ballooning degeneration of liver tissue， alleviation of the compression of hepatic sinusoidal tissue， unobvious inflammatory cell infiltration and fibrous tissue proliferation， reduction in the expression of SREBP1 and PPARγ （P<0.05， P<0.01）， and rise of the protein expression of PPARα （P<0.01）， CLOCK， BMAL1， REV-ERBα， and REV-ERBβ （P<0.05， P<0.01）. ConclusionJTXZT can significantly alleviate the metabolic dysfunction-associated fatty liver disease in mice caused by high-fat diet. The mechanism is the likelihood that it regulates downstream related lipid metabolism proteins （such as SREBP1， PPARγ， and PPARα）.

Effect of Jiangtang Xiaozhi Tablet on Liver Circadian Clock-related Genes CLOCK， BMAL1， REV-ERBα， and REV-ERBβ in Mice with Metabolic Dysfunction-associated Fatty Liver Disease / 中国实验方剂学杂志

ObjectiveTo explore the effect of Jiangtang Xiaozhi tablet （JTXZT） on metabolic dysfunction-associated fatty liver disease and to study the mechanism from the perspective of circadian clock-related genes such as circadian locomotor output cycles kaput （CLOCK）， brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 （BMAL1）， reverse-eritroblastosis receptor （REV-ERB）α and β. MethodA total of 50 male SPF C57BL/6J mice were randomized into normal group （n=10） and modeling group （n=40）. The normal group was fed with normal diet， and the modeling group with high-fat diet for 4 weeks. Then the model mice were randomly classified into model group， high-dose （12.5 g·kg-1） and low-dose （6.25 g·kg-1） Jiangtang Xiaozhi tablet groups， and orlistat group （70 mg·kg-1）， with 10 mice in each group. The normal group and model group received equivalent volume of distilled water （8 weeks）. Then， the body weight of mice was measured， and the content of serum triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） was determined with biochemical method. Serum content of free fatty acid （FFA） and leptin was detected by enzyme-linked immunosorbent assay （ELISA）. Pathological changes of liver tissue and epididymal adipose tissue were observed based on hematoxylin-eosin （HE） staining. Liver fibrosis was examined based on Masson's trichrome staining， and changes of lipids based on oil red O staining. The expression of CLOCK， BMAL1， REV-ERBα， and REV-ERBβ was detected by Western blot and immunohistochemistry assay. ResultCompared with the normal group， the model group had high content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin （P<0.05， P<0.01）， showed ballooning degeneration and focal microvesicular steatosis of liver cells， enlarged adipocytes， and inflammatory cell clusters and fibrous tissue hyperplasia， and displayed increased protein expression of sterol regulatory element binding protein （SREBP） 1 and peroxisome proliferators-activated receptor （PPAR）γ （P<0.01） and decreased protein expression of PPARα （P<0.05）， CLOCK， BMAL1， REV-ERBα and β （P<0.05， P<0.01）. Compared with the model group， JTXZT-H group down-regulated the content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin in mice （P<0.05， P<0.01）， and the JTXZT groups demonstrated reduction in the degree and range of ballooning degeneration of liver tissue， alleviation of the compression of hepatic sinusoidal tissue， unobvious inflammatory cell infiltration and fibrous tissue proliferation， reduction in the expression of SREBP1 and PPARγ （P<0.05， P<0.01）， and rise of the protein expression of PPARα （P<0.01）， CLOCK， BMAL1， REV-ERBα， and REV-ERBβ （P<0.05， P<0.01）. ConclusionJTXZT can significantly alleviate the metabolic dysfunction-associated fatty liver disease in mice caused by high-fat diet. The mechanism is the likelihood that it regulates downstream related lipid metabolism proteins （such as SREBP1， PPARγ， and PPARα）.

Application of prognostic model of fatty acid metabolismrelated genes in renal cell carcinoma / 西安交通大学学报(医学版)

【Objective】 To establish a prognostic model of fatty acid metabolism related genes for predicting the prognosis of renal clear cell carcinoma. 【Methods】 The differentially expressed fatty acid metabolism-related genes in renal clear cell carcinoma samples and normal samples in TCGA database were screened by R language software. The Cox proportional hazard regression model was used to select and establish a multigene prognostic model and the prognostic score was calculated. Patients were divided into high-risk group and low-risk group according to the median prognostic score. Kaplan-Meier survival curve was used to analyze the difference in two groups. The clinical pathological factors and prognostic score factors were included in the Cox regression model to analyze the factors affecting the survival of patients with renal clear cell carcinoma. ROC receiver operating curve analysis was used to evaluate the accuracy of the prognostic prediction model. The prognostic model of fatty acid metabolism-related genes and their correlation with clinical factors were analyzed. GSEA enrichment analysis analyzed the differences of gene sets in risk groups. 【Results】 A total of 4 differential genes (CPT1B, HADH, CYP4A11, and ACADSB) were selected to establish a prognostic model for genes related to fatty acid metabolism in renal cell carcinoma. The prognostic risk score (RS) formula is as follows: RS=0.490×CPT1B-0.428×HADH-0.11 × CYP4A11-0.372 × ACADSB. Kaplan-Meier survival analysis confirmed that the overall survival rate of patients with low-risk prognostic score was significantly higher in patients with overall renal clear cell carcinoma, and the difference was statistically significant (P<0.001). Cox regression analysis showed that the prognostic model of genes related to age and fatty acid metabolism is an independent influencing factor for the prognosis of patients with renal clear cell carcinoma (P<0.01). The 5 years’ AUC of the renal clear cell carcinoma ROC curve of the renal cancer fatty acid metabolism related gene model was 0.802. GSEA analysis showed that the difference of 81 gene sets in the low-risk group was statistically significant (P<0.05). 【Conclusion】 The prognostic model of renal cancer fatty acid metabolism-related genes can be used to predict the prognosis of patients with renal clear cell carcinoma, which is conducive to further guide clinical treatment.

Research progress of the relationship between autophagy and inflammatory bowel disease / 中国临床药理学与治疗学

Autophagy is regulated by autophagy-related genes (ATGs), so that the aged and damaged cellular substances in cells are sent to lysosomes for degradation and recycling, thereby maintaining cellular homeostasis. Inflammatory bowel disease (IBD) is an autoimmune disease, and the ability of autophagy to selectively target intracellular pathogens for destruction is considered a key aspect of the innate immune response. Genetic studies suggest that several autophagy-related genes are clinically relevant in the pathogenesis of IBD. This article will describe the role of autophagy in IBD and the latest advances in its clinical relevance, as well as autophagy-targeted therapeutic strategies for IBD.

Expression of miR-378 in cervical cancer and its effects on the proliferation and invasion of cancer cells as well as the underlying mechanism / 中国基层医药

Objective:To detect the expression of miR-378 in cervical cancer and investigate its effects on the proliferation and invasion of cancer cells as well as the underlying mechanism.Methods:A total of 185 cervical tissue samples of women who received gynecological examination in Qilu Hospital of Shandong University from January 2012 to January 2016 were included in this study. Reverse transcription-quantitative polymerase chain reaction was performed to determine the expression of miR-378 in cervical tissue and C-33A cells. Western blot assay was performed to detect the expression of different cancer genes ATG12, CCND1 and pRb in C-33A cells. BrdU cell proliferation and Transwell invasion assay were performed to determine cell proliferation and invasion. Target Scan was used to predict and screen miR-378 gene targets and verified by a dual-luciferase reporter assay system.Results:The expression of miR-378 in cervical intraepithelial neoplasia (CIN) III lesioned tissue and cervical cancer tissue was significantly higher than that in normal cervical tissues ( F = 103.091, t = 9.381, 8.936, both P < 0.05). The expression of miR-378 in cervical cancer tissues with positive lymph node metastasis was significantly higher than that in cervical cancer tissues with negative lymph node metastasis ( t = 1.007, P < 0.01). The overexpression of miR-378 in cervical cancer tissues significantly promoted the migration and invasion of C-33A cells ( t = 5.285, P < 0.05), while low expression of miR-378 in cervical cancer tissues significantly inhibited the migration and invasion of HeLa cells ( t = 2.941, P < 0.05). The overexpression of miR-378 in C-33A cells significantly decreased the expression of ATG12, CCND1and pRb ( t = 1.382, 1.431 and 2.086, all P < 0.05). The low expression of miR-378 in C-33A cells significantly increased the expression of ATG12, CCND1 and pRb ( t = 3.961, 3.062 and 2.894, all P < 0.05). Conclusion:miR-378 can greatly promote the metastasis of cervical cancer cells. ATG12, as a direct target of miR-378, provides new insights into the molecular mechanism underlying cervical cancer pathology and therapeutic target.

Effects of the ITPR1 gene overexpression on Ca²⁺ concentration, lipid content and calcium transport-related genes in duck uterine epithelial cells / 生物工程学报

Inositol 1,4,5-trisphosphate receptor 1 (ITPR1) is an important intracellular channel for releasing Ca²⁺. In order to investigate the effects of the ITPR1 overexpression on Ca²⁺ concentration and lipid content in duck uterine epithelial cells and its effects on calcium transport-related genes, the structural domain of ITPR1 gene of duck was cloned into an eukaryotic expression vector and transfected into duck uterine epithelial cells. The overexpression of the ITPR1 gene, the concentration of Ca²⁺, the lipid content, and the expression of other 6 calcium transport-related genes was determined. The results showed that the concentration of Ca²⁺ in uterine epithelial cells was significantly reduced after transfection (P<0.05), the triglyceride content was significantly increased (P<0.01), and the high-density lipoprotein content was significantly decreased (P<0.01). The correlation analysis results showed that the overexpression of the C-terminal half of the ITPR1 gene was significantly positively correlated with the total cholesterol content (P<0.01), which was significantly positively correlated with the low-density lipoprotein content (P<0.05). The overexpression of the N-terminal half of the ITPR1 gene was significantly positively correlated with the triglyceride content (P<0.01), which was significantly negatively correlated with the concentration of Ca²⁺ (P<0.05). RT-qPCR results of 6 calcium transport-related genes showed that the overexpression of the C-terminal half of the ITPR1 gene significantly inhibited the expression of the IP3R2, VDAC2 and CAV1 genes, and the overexpression of the N-terminal half of the ITPR1 gene significantly promoted the expression of the IP3R3 and CACNA2D1 genes. In conclusion, the ITPR1 gene overexpression can promote Ca²⁺ release in duck uterus epithelial cells, promote the synthesis of triglyceride, low-density lipoprotein and cholesterol, and inhibit the production of high-density lipoprotein, and the ITPR1 gene overexpression affected the expression of all 6 calcium transport-related genes.

Recommendations for the use of the acetaminophen hepatotoxicity model for mechanistic studies and how to avoid common pitfalls

Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, which is safe at therapeutic doses but can cause severe liver injury and even liver failure after overdoses. The mouse model of APAP hepatotoxicity recapitulates closely the human pathophysiology. As a result, this clinically relevant model is frequently used to study mechanisms of drug-induced liver injury and even more so to test potential therapeutic interventions. However, the complexity of the model requires a thorough understanding of the pathophysiology to obtain valid results and mechanistic information that is translatable to the clinic. However, many studies using this model are flawed, which jeopardizes the scientific and clinical relevance. The purpose of this review is to provide a framework of the model where mechanistically sound and clinically relevant data can be obtained. The discussion provides insight into the injury mechanisms and how to study it including the critical roles of drug metabolism, mitochondrial dysfunction, necrotic cell death, autophagy and the sterile inflammatory response. In addition, the most frequently made mistakes when using this model are discussed. Thus, considering these recommendations when studying APAP hepatotoxicity will facilitate the discovery of more clinically relevant interventions.

Natural compounds modulate the autophagy with potential implication of stroke

Stroke is considered a leading cause of mortality and neurological disability, which puts a huge burden on individuals and the community. To date, effective therapy for stroke has been limited by its complex pathological mechanisms. Autophagy refers to an intracellular degrading process with the involvement of lysosomes. Autophagy plays a critical role in maintaining the homeostasis and survival of cells by eliminating damaged or non-essential cellular constituents. Increasing evidence support that autophagy protects neuronal cells from ischemic injury. However, under certain circumstances, autophagy activation induces cell death and aggravates ischemic brain injury. Diverse naturally derived compounds have been found to modulate autophagy and exert neuroprotection against stroke. In the present work, we have reviewed recent advances in naturally derived compounds that regulate autophagy and discussed their potential application in stroke treatment.

Construction and Validation of Prognostic Risk Score Model of Autophagy Related Genes in Lung Adenocarcinoma / 中国肺癌杂志

BACKGROUND@#Autophagy related genes (ARGs) regulate lysosomal degradation to induce autophagy, and are involved in the occurrence and development of a variety of cancers. The expression of ARGs in tumor tissues has a great prospect in predicting the survival of patients. The aim of this study was to construct a prognostic risk score model for lung adenocarcinoma (LUAD) based on ARGs.@*METHODS@#5,786 ARGs were obtained from GeneCards database. Gene expression profiles and clinical data of 395 LUAD patients were collected from The Cancer Genome Atlas (TCGA) database. All ARGs expression data were extracted, and The ARGs differentially expressed were identified by R software. Survival analysis of differentially expressed ARGs was performed to screen for ARGs with prognostic value, and functional enrichment analysis was performed. The least absolute selection operator (LASSO) regression and Cox regression model were used to construct a prognostic risk scoring model for ARGs. The receiver operating characteristic (ROC) curve was drawn to obtain the optimal cut-off value of risk score. According to the cut-off value, the patients were divided into high-risk group and low-risk group. The area under curve (AUC) and the Kaplan-Meier survival curve was plotted to evaluate the model performance, which was verified in external data sets. Finally, univariate and multivariate Cox regression analysis was applied to evaluate the independent prognostic value of the model, and its clinical relevance was analyzed.@*RESULTS@#Survival analysis, Lasso regression and Cox regression analysis were used to construct a LUAD prognostic risk score model with five ARGs (ADAM12, CAMP, DKK1, STRIP2 and TFAP2A). The survival time of patients with low-risk score in this model was significantly better than that of patients with high-risk score (P<0.001). The model showed good prediction performance for LUAD in both the training set (AUCmax=0.78) and two external validation sets (AUCmax=0.88). Risk score was significantly associated with the prognosis of LUAD patients in univariate and multivariate Cox regression analyses, suggested that risk score could be a potential independent prognostic factor for LUAD. Correlation analysis of clinical characteristic showed that high risk score was closely associated with high T stage, high tumor stage and poor prognosis.@*CONCLUSIONS@#We constructed a LUAD risk score model consisting of five ARGs, which can provide a reference for predicting the prognosis of LUAD patients, and may be used in combination with tumor node metastasis (TNM) staging for prognosis prediction of LUAD patients in the future.

Effect of Qiyu Sanlong Decoction in Inhibiting Growth of Lung Cancer by Regulating Expressions of Beclin1, Atg5 and LC3B / 中国实验方剂学杂志

Objective::To study the effect of Qiyu Sanlong decoction on the growth of subcutaneous tumor in lung cancer mice and the expressions of key autophagy molecule, yeast Atg6 homologous (Beclin1), autophagy related genes5 (Atg5), and microtubule-associated protein1 light chain3 (LC3B). Method::Lewis lung carcinoma cells (LLC) were used to reproduce the lung cancer mice transplanted model. After the modeling, the mice were randomly divided into model group, Qiyu Sanlong decoction group, chemotherapy group and combination group, with 18 transplanted mice in each group. In model group, mice were fed with 0.9% saline 20 mL·kg-1 daily. In Qiyu Sanlong decoction group, mice were fed with Qiyu Sanlong decoction 80.48 g·kg-1 daily. The chemotherapy group was intraperitoneally injected with 0.4 mL cisplatin solution (DDP) at the 1st, 3rd and 5th day. The combination group was orally given the drugs at the concentration of 80.48 g·kg-1, and 0.4 mL DDP solution was intraperitoneally injected at the 1st, 3rd and 5th day. After 21 days of continuous treatment, tumor tissue was exfoliated and weighed, and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the histological changes of tumor. The expressions and localizations of Beclin1 and LC3B in tumor tissues were detected by immunohistochemical staining. Protein expressions of Beclin1, Atg5, LC3B-Ⅰand LC3B-Ⅱ were determined by Western blot, and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated. The transcription levels of Beclin1, Atg5 mRNA in tumor tissues were detected by Real-time PCR. Result::Qiyu Sanlong decoction had a mild inhibitory effect on transplanted tumor, with an inhibitory rate of 31.2%. Under microscope, patchy necrotic tumor cells were observed in the tumor tissues of Qiyu Sanlong decoction group. Immunohistochemical staining and Western blot analysis showed that Qiyu Sanlong decoction could up-regulate the expressions of Beclin1, Atg5 and LC3B protein (P<0.01), and promote the conversion from LC3B-Ⅰ into LC3-Ⅱ compared with the model group. Real-time PCR results showed that Qiyu Sanlong decoction could promote the transcription of Beclin1 mRNA and Atg5 mRNA compared with the model group (P<0.01). Conclusion::Qiyu Sanlong decoction has a mild inhibitory effect on lung tumors, and its mechanism may be related to up-regulating the expressions of autophagy key proteins Beclin1, Atg5 and LC3B, and promoting the conversion from LC3B-Ⅰ to LC3B-Ⅱ.

Effects of strontium-modified titanium surfaces on adhesion, migration and proliferation of bone marrow mesenchymal stem cells and expression of bone formation-related genes / 中国组织工程研究

BACKGROUND: Strontium promotes bone formation, and reduces bone resorption. Strontium-modified titanium implant surface has been the focus of research in implant osseointegration under osteoporotic conditions. OBJECTIVE: To investigate the cell adhesion, stretch, and migration of advanced strontium-modified titanium surfaces using bone marrow mesenchymal stem cells isolated from rats. METHODS: Strontium-modified titanium surfaces were produced by sequential treatments with 5 mol/L NaOH, 100 mmol/L strontium acetate, and heated at 600 or 700 °C for 1 hour. After heat treatment, half the samples were soaked in deionized water at 80 °C for 24 hours, then washed and dried. The pure titanium served as control group, and there were five groups. The whole bone marrow adherence method was used to separate bone marrow mesenchymal stem cells from the rats. The modified titanium sheets were placed in 24-well plates and cultured in cell suspension. Cell adhesion and cell proliferation were assessed using cell counting kit-8 assay. After bone marrow mesenchymal stem cells were with the titanium sheet for 24 hours, the actin was stained to observe cell adhesion and stretch. The migration of bone marrow mesenchymal stem cells on different titanium surfaces was investigated using cell scratch test and fluorescence staining. The expression levels of collagen type І, Runx2, osteoprotegerin, RANKL and osteopontin were detected by qRT-PCR. RESULTS AND CONCLUSION: Strontium-modified titanium could promote the stretch and migration of bone marrow mesenchymal stem cells. The number of proliferative cells in the Sr700 group was significantly higher than that in the Sr600 group at 5 days (P < 0.01). On day 14, strontium-modified titanium promoted the expression of collagen type І, Runx2, and osteopontin. In summary, strontium-modified titanium can promote adhesion, spreading, migration and proliferation of rat bone marrow mesenchymal stem cells. Moreover, hydration treatment can enhance the osteogenic activity of rat bone marrow mesenchymal stem cells.

Autophagy and epigenetic modification in inflammatory bowel disease / 中国组织工程研究

BACKGROUND: Inflammatory bowel disease is a chronic inflammatory disease associated with intestinal immune, and autophagy is a cell approach to promote immune regulation. Abnormal expression of autophagy-related genes is closely related to intestinal inflammation and immune response. However, the mechanism by which epigenetic modification regulates autophagy in inflammatory bowel disease has not been fully clarified. OBJECTIVE: To introduce the role of epigenetic modification in autophagy, and to promote a further understanding of inflammatory bowel disease. METHODS: A computer-based online research of PubMed database was performed with the key words of “inflammatory bowel disease, autophagy, autophagy-related genes, epigenetic modification, DNA methylation, histone modification, chromatin remodeling, miRNA.” The search time was from January 1998 to April 2019. Finally, 61 eligible articles were included in result analysis. RESULTS AND CONCLUSION: Epigenetic modifications such as DNA methylation, histone modification, chromatin remodeling, non-coding RNA can regulate intestinal inflammation, immune and autophagy through susceptibility genes AGL and IRGM, thereby mediating the occurrence and development of inflammatory bowel disease.

Expression of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients / 中国组织工程研究

BACKGROUND: Bones are currently considered as an immune organ. A variety of immune cells that originate from the bone marrow can interact with the cells of the skeletal system to jointly regulate bone metabolism. Explorations on the pathogenesis of postmenopausal osteoporosis as well as treatment-related molecular targets and signal pathways can help prevention and treatment of the disease. OBJECTIVE: To investigate the expression profiles of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients using RNA-Seq technology. METHODS: Forty female patients who had experienced menopause for 0 to 20 years and were hospitalized due to fractures were enrolled. They were divided into normal bone mass group (T >-1) and osteoporosis group (T 2), and 131 genes were up-regulated and 56 genes were down-regulated. We identified in total 29 differentially expressed immune-related genes including 25 up-regulated and 4 down-regulated ones. There was significant difference in expression between the osteoporosis and normal bone mass groups for genes, including KIR3DL1, KIR3DL2, KIR2DL4, KLRD1 and HSPA6 (P < 0.05). These differentially expressed genes are potentially important for the natural killer cell-mediated cytotoxicity by the KEGG pathway analysis. KIR3DL1, KIR3DL2, KIR2DL4, KLRD1 and HSPA6 may be closely related to the natural killer cell-mediated cytotoxicity during the occurrence of postmenopausal osteoporosis.