Inhibition of NLRP3 Inflammasome Ameliorates Cerebral Ischemia-Reperfusion Injury in Diabetic Mice.

Sustained activation of NLRP3 inflammasome is closely related to diabetes and stroke. However, it is unknown whether NLRP3 inflammasome plays an essential role in stroke in diabetes. We aim to investigate the effect and the potential mechanism of NLRP3 inflammasome in diabetic mice with cerebral ischemia-reperfusion injury. A type 2 diabetic mouse model was induced by a high-fat diet and streptozotocin (STZ). Diabetic mice received MCC950 (the specific molecule NLRP3 inhibitor) or vehicle 60 minutes before the middle cerebral artery occlusion (MCAO) and reperfusion. MCC950 reduced the neurological deficit score of 24 h after cerebral ischemia reperfusion and improved the 28-day survival rate of cerebral ischemia-reperfusion injury in diabetic mice. Furthermore, we found that the mRNA transcription levels of NLRP3, IL-1ß, and caspase-1 in the core ischemic area were remarkably amplified in diabetic mice with cerebral ischemia-reperfusion injury, whereas this phenomenon was obviously attenuated by MCC950 pretreatment. In conclusion, the NLRP3 inflammasome was involved in the complex diseases of diabetic stroke. MCC950, the NLRP3 specific inhibitor, ameliorated diabetic mice with cerebral ischemia-reperfusion injury and improved the 28-day survival rate during the recovery stage of ischemic stroke.

[The Relationship of Polymorphisms in Interleukin 17A Gene, Interleukin 17F Gene and the Pulmonary Inflammation Risk in Dust Exposed Workers in a Chinese Population].

OBJECTIVES: To investigate the polymorphisms in interleukin 17A IL-17A) and interleukin 17F (IL-17F) and their relationship with pulmonary inflammation risk of dust exposed workers. METHODS: A case-control study among 193 subjects, including 67 subjects in case group and 126 in control group was conducted. PCR-RFLP was applied to genotype IL-17A (G-197A) and IL-17F (7488T/C). Logistic regression analysis was used to determine the effects of IL-17A (G-197A) and IL-17F (7488T/C) on the lung inflammation risk in dust exposed workers. RESULTS: The genotypes analysis showed that the proportions of IL-17A (G-197A) A/A, A/G and G/G were 42 (21.76%), 95 (49.22%), 56 (29.02%) in 193 cases, respectively, and the IL-17F (7488T/C) T/T, T/C and C/C genotypes were 128 (66.32%), 54 (28.98%), 11 (5.70%), respectively. The frequency distribution of each genotype was consistent with the Hardy-Weinberg equilibrium fixed law. The ratio of IL-17A (G-197A) A/A in the case group was lower than that of control group ( P<0.05), while the G/G and A/G genotypes were higher than that of control group ( P<0.05). Furthermore, the genotypes of IL-17A (G-197A) A/G (OR=5.03, P<0.01) and G/G(OR=3.35, P<0.05) were associated with an increased risk of lung inflammation in workers exposed to dust. The frequency distribution difference of IL-17F (7488T/C) T/T, T/C and C/C genotypes in the cases and control group was unobvious ( P>0.05). CONCLUSIONS: Under the same dust concentration, the dust exposed workers carrying IL-17A (G-197A) A/G, G/G genotypes are more susceptible to pulmonary inflammation in the southwest of China.

Liver receptor homologue-1 and steroidogenic factor-1 expression in cultured granulosa cells from patients with endometriosis: A preliminary study.

AIM: The expression of aromatase (via CYP19 and the CYP19 PII promoter) and the orphan nuclear receptor family members, liver receptor homologue-1 (LRH-1) and steroidogenic factor-1 (SF-1) in cultured luteinized granulosa cells from women with endometriosis were investigated. METHODS: Luteinized granulosa cells from patients undergoing in vitro fertilization (16 patients with endometriosis and 28 controls) were examined for messenger ribonucleic acid (mRNA) expression of CYP19, CYP19 PII, LRH-1 and SF-1, determined by quantitative reverse transcription-polymerase chain reaction. Student's t-test, the Mann-Whitney U test and Spearman's rank correlation were used for statistical analysis. RESULTS: The number of high quality embryos in the endometriosis group was significantly lower than in the control group. The mRNA expression levels of CYP19, CYP19 PII, LRH-1 and SF-1 in granulosa-lutein cells were decreased in women with endometriosis compared to the control group. The simultaneous down-regulation expression of LRH-1, SF-1 and CYP19 PII in endometriotic granulosa cells indicated their positive correlation. CONCLUSION: Our results demonstrate aberrant expressions of SF-1 and LRH-1 in endometriotic granulosa-lutein cells. This finding may be helpful in understanding infertility associated with endometriosis and reduced P450 aromatase activity in endometriotic granulosa cells.

Gut microbiota-derived nicotinamide mononucleotide alleviates acute pancreatitis by activating pancreatic SIRT3 signalling.

BACKGROUND AND PURPOSE: Gut microbiota dysbiosis induced by acute pancreatitis (AP) exacerbates pancreatic injury and systemic inflammatory responses. The alleviation of gut microbiota dysbiosis through faecal microbiota transplantation (FMT) is considered a potential strategy to reduce tissue damage and inflammation in many clinical disorders. Here, we aim to investigate the effect of gut microbiota and microbiota-derived metabolites on AP and further clarify the mechanisms associated with pancreatic damage and inflammation. EXPERIMENTAL APPROACH: AP rat and mouse models were established by administration of caerulein or sodium taurocholate in vivo. Pancreatic acinar cells were exposed to caerulein and lipopolysaccharide in vitro to simulate AP. KEY RESULTS: Normobiotic FMT alleviated AP-induced gut microbiota dysbiosis and ameliorated the severity of AP, including mitochondrial dysfunction, oxidative damage and inflammation. Normobiotic FMT induced higher levels of NAD+ (nicotinamide adenine dinucleotide)-associated metabolites, particularly nicotinamide mononucleotide (NMN). NMN administration mitigated AP-mediated mitochondrial dysfunction, oxidative damage and inflammation by increasing pancreatic NAD+ levels. Similarly, overexpression of the NAD+ -dependent mitochondrial deacetylase sirtuin 3 (SIRT3) alleviated the severity of AP. Furthermore, SIRT3 deacetylated peroxiredoxin 5 (PRDX5) and enhanced PRDX5 protein expression, thereby promoting its antioxidant and anti-inflammatory activities in AP. Importantly, normobiotic FMT-mediated NMN metabolism induced SIRT3-PRDX5 pathway activation during AP. CONCLUSION AND IMPLICATIONS: Gut microbiota-derived NMN alleviates the severity of AP by activating the SIRT3-PRDX5 pathway. Normobiotic FMT could be served as a potential strategy for AP treatment.

TNF-α and IL-1RA polymorphisms and silicosis susceptibility in Chinese workers exposed to silica particles: a case-control study.

OBJECTIVE: To assess the association of TNF-α and IL-1RA SNPs with the risk of silicosis in Chinese workers exposed to silica particles. METHODS: Case-control study design was used to enroll 68 silicotic patients induced by silica particles and 68 healthy workers matched for length of silica particle exposure as controls. Both cases and controls were from the same company in southwest China, and each of them was requested to complete a questionnaire. Blood samples were drawn for genomic DNA extraction from each participant. The genotyping of TNF-α (-238 and -308) and IL-1RA (+2018) was performed using polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) and SYBR green-based quantitative polymerase chain reaction (qPCR), respectively. Unconditional logistic regression model was used to estimate odds ratios (ORs) and their 95% confidential intervals (CI) for SNPs. RESULTS: No significant differences were found between cases and controls in particles exposure length, body mass index (BMI), and status of smoking and alcohol consumption except for age (P=0.001) and blood type (P=0.042). The frequencies of TNF-α (-238) and IL-1RA (+2018) genotypes in cases were significantly different from those in controls, (P=0.001 and P=0.002, respectively), while a borderline significant difference was found in the frequencies of TNF-α (-308) between cases and controls (P=0.063). The variants of three SNPs increased the risk of silicosis in the Chinese workers exposed to silica particles. The adjusted ORs of TNF-α (-308), TNF-α (-238) and IL-1RA (+2018) were 2.8 (95% CI: 1.1-7.5), 20.9 (95% CI: 1.8-236.4) and 4.0 (95% CI: 1.6-10.1), respectively. CONCLUSION: It is suggested that cytokine polymorphisms of TNF-α (-238, -308) and IL-1RA (+2018) are associated with the risk of silicosis in the Chinese workers exposed to silica particles. Further independent studies on the interaction between SNPs and exposure to silica particles with a larger sample size are therefore warranted.

Chemical composition and antimicrobial activity of the essential oil of kumquat (Fortunella crassifolia Swingle) peel.

The aim of this study was to determine the main constituents of the essential oil isolated from Fortunella crassifolia Swingle peel by hydro-distillation, and to test the efficacy of the essential oil on antimicrobial activity. Twenty-five components, representing 92.36% of the total oil, were identified by GC-MS analysis. The essential oil showed potent antimicrobial activity against both Gram-negative (E. coli and S. typhimurium) and Gram-positive (S. aureus, B. cereus, B. subtilis, L. bulgaricus, and B. laterosporus) bacteria, together with a remarkable antifungal activity against C. albicans. In a food model of beef extract, the essential oil was observed to possess an effective capacity to control the total counts of viable bacteria. Furthermore, the essential oil showed strongly detrimental effects on the growth and morphological structure of the tested bacteria. It was suggested that the essential oil from Fortunella crassifolia Swingle peel might be used as a natural food preservative against bacteria or fungus in the food industry.

[Short-term medication of L-carnitine before intracytoplasmic sperm injection for infertile men with oligoasthenozoospermia].

OBJECTIVE: To observe the pregnancy promoting effect of L-carnitine combined with intracytoplasmic sperm injection (ICSI) in treating male infertility with oligoasthenozoospermia. METHODS: We assigned 129 patients with oligoasthenozoospermia to receive 2 weeks of oral L-carnitine followed by ICSI (medication group, n = 42) and ICSI alone (control group, n = 87). We compared the sperm concentration and motility, the percentage of grade a + b sperm, and sperm deformity before and after L-carnitine medication, as well as the rates of fertilization, cleavage, available embryo and clinical pregnancy between the two groups. RESULTS: The percentage of grade a + b sperm was significantly increased after L-carnitine medication as compared with the baseline ([13.5 +/- 10.7] % vs [9.6 +/- 7.2] %, P<0.05), and so was the rate of available embryo in the medication group after ICSI in comparison with that of the control group (77.50% vs 69.04%, P<0.05). CONCLUSION: Short-term medication of L-carnitine can improve sperm quality and raise the success rate of ICSI.

Gut microbiota patterns associated with somatostatin in patients undergoing pancreaticoduodenectomy: a prospective study.

Postoperative pancreatic fistula (POPF) is a common and dreaded complication after pancreaticoduodenectomy (PD). The gut microbiota has been considered as an crucial mediator of postoperative complications, however, the precise roles of gut microbiota in POPF are unclear. A prospective study was developed to explore the effects of somatostatin on gut microbiota and we aim to identify the microbial alterations in the process of POPF. A total of 45 patients were randomly divided into PD group or additional somatostatin therapy group. The fecal sample of each patient was collected preoperatively and postoperatively and the gut microbiota was analyzed by 16S rRNA sequencing. Our study found that somatostatin therapy was independent risk factor for the occurrence of POPF, and it reduced the microbial diversity and richness in patients. At genus level, somatostatin therapy led to a decreased abundance in Bifidobacterium, Subdoligranulum and Dubosiella, whereas the abundance of Akkermansia, Enterococcus and Enterobacter were increased. The abundance levels of certain bacteria in the gut microbiota have significantly shifted in patients with POPF. The LEfSe analysis revealed that Ruminococcaceae could be used as microbial markers for distinguishing patients with high risk of POPF. Furthermore, Verrucomicrobia and Akkermansia could be used as preoperative biomarkers for identifying patients without POPF. Our prospective study highlights the specific communities related with somatostatin therapy and discovers POPF-associated microbial marker, which suggests that gut microbiota may become a diagnostic biomarker and potential therapeutic target for POPF.

Correction: Gut microbiota patterns associated with somatostatin in patients undergoing pancreaticoduodenectomy: a prospective study.

[This corrects the article DOI: 10.1038/s41420-020-00329-4.].

The influence of early cleavage on embryo developmental potential and IVF/ICSI outcome.

PURPOSE: To observe whether early cleavage can be a predictor of embryo developmental potential, pregnancy and implantation rates. METHODS: A total of 9,544 embryos in 1,095 in vitro fertilization or intracytoplasmic sperm injection cycles were observed with regard to the appearance of early cleavage at 25-29 h post-insemination. RESULTS: A significantly higher proportion of excellent quality embryos were observed in the early cleavage group compared to the late cleavage group (52.5 versus 28.9%, P < 0.01). In the early cleavage group there was also a higher rate of pregnancy per transfer compared with the late cleavage group (38.7 versus 26.3%, P < 0.01). In addition, we found that transfer of only one early cleavage embryo resulted in a high pregnancy rate (38.5%) and a low multiple pregnancy rate (18.0%). CONCLUSION: Early cleavage is a strong indicator of embryo quality, and may be used as an additional criterion in the selection of embryos for transfer to increase pregnancy rate and reduce multiple pregnancy rate.

[Inhibitory effect of N-acetylcysteine upon atherosclerotic processes in rabbit carotid].

OBJECTIVE: To discuss the effect of N-acetylcysteine (NAC) upon matrix metalloproteinases (MMP) in the atherosclerotic processes in rabbit carotid. METHODS: The atherosclerotic models were generated in vitro by injuring rabbit internal carotid with arterial canal balloon. These rabbits were divided into 3 groups (15 mg/kg NAC, 30 mg/kg NAC and control group) and treated for 8 weeks. HE staining and immunohistochemistry were used to observe the plaque formation and the distribution of MMPs and ox-LDL. ELISA was used to detect the level of ox-LDL. And the protein levels of MMP-2 and MMP-9 in rabbit venous blood were detected by SDS PAGE zymography. The mRNA level of MMP-2 and MMP-9 were measured by RT-PCR and electrophoresis. RESULTS: As compared with the control group, NAC (15 mg/kg) group had a reduction of neointima of arterial lumen [(1.79 +/- 0.24) vs (2.78 +/- 0.17) mm2]. A decrease of endothelial thickness [(0.16 +/- 0.01) vs (0.24 +/- 0.02) mm2] and an increase of vascular cavity transverse [(0.58 +/- 0.10) vs (0.33 +/- 0.1) mm2] (P < 0.05) were observed. At week 8, the oxLDL levels decreased by 16% in NAC (30 mg/kg) group [(30.5 +/- 1.2) vs (36.2 +/- 1.8) mmol/L] (P < 0.01). Serum levels of pro-MMP-2, MMP-2 and pro-MMP-9 decreased markedly [INT/mm2: (311 +/- 19, 208 +/- 8, 283 +/- 7 vs 619 +/- 17, 574 +/- 8, 564 +/- 10) respectively, P < 0.01] in NAC (30 mg/kg) group. The levels of mRNA expression of MMP-2 and MMP-9 were (2.4 +/- 0.4, 2.8 +/- 0.2) vs (3.4 +/- 0.3, 3.7 +/- 0.5) respectively (P < 0.05). CONCLUSION: NAC inhibits the atherosclerotic formation, suppresses the levels of ox-LDL, MMP-9 and MMP-2 and downgrades the expression of matrix metalloproteinase mRNA.

[Important Effect of Secondary Inorganic Salt Extinction on Visibility Impairment in the Northern Suburb of Nanjing].

Observed data regarding the visibility and aerosol chemical composition from May 2013 to May 2014 were used to analyze the variation of visibility, the relationship between aerosol chemical composition and visibility variations, and their contributions to atmospheric light extinction. An important effect of secondary inorganic salt extinction on the visibility impairment was determined. The present study suggests that the average visibility during the observation period was (6.78±3.68) km, and there was obvious seasonal variation in the visibility. Fine particles with size less than 2.1 µm have a great influence on visibility, with the main chemical components of SO42-, NO3-, NH4+, and OC. The secondary inorganic ions make significant contributions to visibility degradation. The mean light extinction coefficient of Nanjing was (527.2±295.2) Mm-1, which was calculated by using the revised IMPROVE equation. Regarding the chemical composition of PM2.1, the most contributive species to the light extinction coefficient were ammonium sulfate, ammonium nitrate, and organic species, which accounted for 80.6%. Although the light extinction contribution of organic matter was as high as 43.51% on a clear day (VR>10 km), with the decrease of visibility, the extinction contribution of organic matter decreased, but the contribution of secondary inorganic salt increased. The contribution of extinction was 58.96% for heavy haze days with low visibility (VR<5 km). This proves that the secondary inorganic salt extinction plays a significant role in visibility impairment.

Isolation and identification of bovine nasopharyngeal mucosal epithelial cells and establishment of cell models of acute infection by foot-and-mouth disease virus.

Foot-and-mouth disease (FMD) commonly occurs via the respiratory tract, and bovine nasopharyngeal mucosal epithelial cells are the primary infection cells in cattle. The aim of the present study was to isolate and culture epithelial cells from the bovine nasopharyngeal mucosa in vitro using a mechanical separation method. The cells were expanded, established in continuous cell culture, and used for immunofluorescence cytochemistry and establishment of infection models. We detected pan-cytokeratin markers of bovine nasopharyngeal mucosal epithelial cells by immunofluorescence. Bovine nasopharyngeal mucosal epithelial cells were then infected with foot-and-mouth disease virus (FMDV) serum type O. RT-PCR demonstrated the successful establishment of acute FMDV infection in the cell models. This infection model provides the basis for clarification of the interaction between FMDV and host bovine nasopharyngeal mucosal epithelial cells in vitro.

Host-guest interaction of ß-cyclodextrin with isomeric ursolic acid and oleanolic acid: physicochemical characterization and molecular modeling study.

Ursolic acid (UA) and oleanolic acid (OA) are insoluble drugs. The objective of this study was to encapsulate them into ß-cyclodextrin (ß-CD) and compare the solubility and intermolecular force of ß-CD with the two isomeric triterpenic acids. The host-guest interaction was explored in liquid and solid state by ultraviolet-visible absorption,1 H NMR, phase solubility analysis, and differential scanning calorimetry, X-ray powder diffractometry, and molecular modeling studies. Both experimental and theoretical studies revealed that ß-CD formed 1: 1 water soluble inclusion complexes and the complexation process was naturally favorable. In addition, the overall results suggested that ring E with a carboxyl group of the drug was encapsulated into the hydrophobic CD nanocavity. Therefore, a clear different inclusion behavior was observed, and UA exhibited better affinity to ß-CD compared with OA in various media due to little steric interference, which was beneficial to form stable inclusion complex with ß-CD and increase its water solubility effectively.

Performance of underground heat storage system in a double-film-covered greenhouse.

An underground heat storage system in a double-film-covered greenhouse and an adjacent greenhouse without the heat storage system were designed on the basis of plant physiology to reduce the energy consumption in greenhouses. The results indicated that the floor temperature was respectively 5.2 degrees C, 4.6 degrees C and 2.0 degrees C higher than that of the soil in the adjacent reference greenhouse after heat storage in a clear, cloudy and overcast sky in winter. Results showed that the temperature and humidity were feasible for plant growth in the heat saving greenhouse.

Identification of FECH gene multiple variations in two Chinese patients with erythropoietic protoporphyria and a review.

Erythropoietic protoporphyria (EPP), an autosomal dominant disease, is caused by partial deficiency of ferrochelatase (FECH), which catalyzes the terminal step of heme biosynthesis because of loss-of-function mutations in the FECH gene. To date, only a few cases have been described in Asia. In this study, we describe the clinical features of two Chinese patients with EPP, with diagnosis confirmed by the increase of free protoporphyrin in erythrocytes, detection of plasma fluorescence peak at 630-634 nm, and analysis of FECH gene mutations. Using gene scanning, we identified a small deletion in the FECH gene (c.973 delA) in one proband (patient A) and a pathogenic FECH mutation (c.1232 G>T) in the other (patient B) and also observed some nucleotide variations (c.798 C>G, c.921 A>G, IVS1-23 C>T, IVS3+23 A>G, IVS9+35 C>T, and IVS3-48 T>C) in these patients. The family pedigree of patient A was then established by characterization of the genotype of the patient's relatives. We also analyzed the potential perniciousness of the missense mutation with bioinformatic software, Polyphen and Sift. In summary, Chinese EPP patients have similar manifestations to those of Caucasians, and identification of the Chinese FECH gene mutations expands the FECH genotypic spectrum and may contribute to genetic counseling.

Long Noncoding RNA MALAT1 Promotes Aggressive Pancreatic Cancer Proliferation and Metastasis via the Stimulation of Autophagy.

Recently, pancreatic ductal adenocarcinoma (PDAC) has emerged as one of the most aggressive malignant tumors with the worst prognosis. Previous studies have demonstrated that long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is increased in pancreatic cancer and is identified as a diagnostic biomarker. Nonetheless, the molecular mechanism of elevated MALAT1 levels and tumor aggressiveness remains unknown. In this study, MALAT1 was found to be highly expressed in PDAC tissues, and elevated expression was associated with poorer prognoses. In addition, MALAT1 was positively linearly correlated with the expression of LC3B mRNA. Furthermore, several molecules involved in cellular autophagic flux were modulated following the downregulation of MALAT1, including LC3, P62, and LAMP-2. Mechanistically, we found that MALAT1 interacted with RNA binding protein HuR, and silencing of MALAT1 greatly enhanced the posttranscriptional regulation of TIA-1 and had further effects on inhibiting autophagy. MALAT1 was speculated to regulate tumorigenesis via HuR-TIA-1-mediated autophagic activation. Hence, we investigated the biological properties of MALAT1 in terms of tumor proliferation and metastasis by promoting autophagy in vitro In brief, these data demonstrate that MALAT1 could facilitate the advanced progression of tumors in vivo Our study highlights the new roles of MALAT1 on protumorigenic functioning and anticancer therapy via activating autophagy in pancreatic cancer. Mol Cancer Ther; 15(9); 2232-43. ©2016 AACR.

Plasma and tumor levels of Linc-pint are diagnostic and prognostic biomarkers for pancreatic cancer.

Long intergenic non-protein coding RNA, p53 induced transcript (Linc-pint) is a long noncoding RNA (lncRNA) that regulates tumor cell viability and proliferation. We used qRT-PCR and RNA FISH analysis to evaluate Linc-pint levels in the plasma and tumor tissues of pancreatic cancer (PCa) patients. Our data demonstrate that Linc-pint expression is lower in plasma samples from PCa patients than from healthy individuals, and indicate that plasma Linc-pint levels are more sensitive than CA19-9 for detecting PCa. Our data also show that Linc-pint levels are lower in PCa tumors than in adjacent tissues, carcinoma of the ampulla of Vater (CAV) and cholangiocarcinoma (CCA), and suggest that Linc-pint could be used for distinguishing the cause of malignant obstructive jaundice. Low plasma Linc-pint levels correlate with tumor recurrence, while low tumor Linc-pint levels correlate with poor prognosis for PCa patients after pancreatectomy. These results thus indicate that low plasma Linc-pint expression could serve as a minimally invasive biomarker for early PCa detection, and that low Linc-pint levels in PCa tumors could be used for predicting patient prognosis.

Protective Effects of Hydrogen Gas on Experimental Acute Pancreatitis.

Acute pancreatitis (AP) is an inflammatory disease mediated by damage to acinar cells and pancreatic inflammation. In patients with AP, subsequent systemic inflammatory responses and multiple organs dysfunction commonly occur. Interactions between cytokines and oxidative stress greatly contribute to the amplification of uncontrolled inflammatory responses. Molecular hydrogen (H2) is a potent free radical scavenger that not only ameliorates oxidative stress but also lowers cytokine levels. The aim of the present study was to investigate the protective effects of H2 gas on AP both in vitro and in vivo. For the in vitro assessment, AR42J cells were treated with cerulein and then incubated in H2-rich or normal medium for 24 h, and for the in vivo experiment, AP was induced through a retrograde infusion of 5% sodium taurocholate into the pancreatobiliary duct (0.1 mL/100 g body weight). Wistar rats were treated with inhaled air or 2% H2 gas and sacrificed 12 h following the induction of pancreatitis. Specimens were collected and processed to measure the amylase and lipase activity levels; the myeloperoxidase activity and production levels; the cytokine mRNA expression levels; the 8-hydroxydeoxyguanosine, malondialdehyde, and glutathione levels; and the cell survival rate. Histological examinations and immunohistochemical analyses were then conducted. The results revealed significant reductions in inflammation and oxidative stress both in vitro and in vivo. Furthermore, the beneficial effects of H2 gas were associated with reductions in AR42J cell and pancreatic tissue damage. In conclusion, our results suggest that H2 gas is capable of ameliorating damage to the pancreas and AR42J cells and that H2 exerts protective effects both in vitro and in vivo on subjects with AP. Thus, the results obtained indicate that this gas may represent a novel therapy agent in the management of AP.

The activation of c-Jun NH2-terminal kinase is required for dihydroartemisinin-induced autophagy in pancreatic cancer cells.

BACKGROUND: c-Jun NH2-terminal kinases (JNKs) are strongly activated by a stressful cellular environment, such as chemotherapy and oxidative stress. Autophagy is a protein-degradation system in which double-membrane vacuoles called autophagosomes are formed. The autophagy-related gene Beclin 1 plays a key role in this process. We previously found that autophagy was induced by dihydroartemisinin (DHA) in pancreatic cancer cells. However, little is known about the complex relationship between ROS, JNK activation, autophagy induction, and Beclin 1 expression. METHODS: Cell viability and CCK-8 assays were carried out to determine the cell proliferation; small interfering RNAs (siRNAs) were used to knockdown c-Jun NH2-terminal kinases (JNK1/2) genes; western blot was performed to detect the protein expression of LC3, JNK, Beclin 1, caspase 3 and ß-actin; production of intracellular ROS was analyzed using FACS flow cytometry; autophagy induction was confirmed by electron microscopy. RESULTS: In the present study, we explored the role of DHA and Beclin 1 expression in autophagy. DHA-treated cells showed autophagy characteristics, and DHA also activated the JNK pathway and up-regulated the expression of Beclin 1. Conversely, blocking JNK signaling inhibited Beclin 1 up-regulation. JNK activation was found to primarily depend on reactive oxygen species (ROS) resulting from the DHA treatment. Moreover, JNK pathway inhibition and Beclin 1 silencing prevented the induction of DHA-induced autophagy. CONCLUSIONS: These results suggest that the induction of autophagy by DHA is required for JNK-mediated Beclin 1 expression.