CircPRKCI regulates proliferation, migration and cycle of lung adenocarcinoma cells by targeting miR-219a-5p-regulated CAMK1D.

OBJECTIVE: Circular ribonucleic acids (circRNAs) are considered as the key regulatory factors for human malignancies in recent years, and lung adenocarcinoma (LUAD) is a common malignancy worldwide, but the molecular mechanism of circRNAs in LUAD has not been completely investigated. Therefore, the mechanism by which circRNA protein kinase C iota (circPRKCI) regulates LUAD cell migration proliferation, and cycle was preliminarily explored in this research, so as to provide new ideas for the treatment of LUAD. PATIENTS AND METHODS: First of all, the circPRKCI expression level in LUAD tissues was tested via quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay, and the relationship between circPRKCI and the patients' prognosis was analyzed. Then, circPRKCI expression was inhibited by small interfering RNA (siRNA), and the influence of circPRKCI on t LUAD cells' ability to proliferate was verified via 5-ethynyl-2'-deoxyuridine (EdU) and cell counting kit-8 (CCK-8) assays. Moreover, the influence of circPRKCI on LUAD cells' ability to migrate was testified by transwell assay, and the regulation of LUAD cell cycle by circPRKCI was confirmed by flow cytometry. The micro RNAs (miRNAs) with binding sites to the 3' untranslated region (UTR) of circPRKCI and the genes binding to miRNAs were discovered using bioinformatics websites, and their associative relation was further explored through Dual-Luciferase reporter gene assay, qRT-PCR assay, Pearson correlation analysis and reverse experiment. RESULTS: It was verified via qRT-PCR assay that circPRKCI was expressed at a remarkably higher level in LUAD tissues relative to that in paracancerous normal tissues. The highly expressed circPRKCI led to poor prognosis of patients. Besides, qRT-PCR assessment results indicated that circPRKCI expression level rose notably in LUAD cell lines, while it was lowered markedly in LUAD cells transfected with si-circPRKCI. According to CCK-8 and EdU assay results, the proliferative ability of LUAD cells was weakened clearly after knocking down circPRKCI. It was manifested in the results of transwell assay that the knockdown of circPRKCI significantly repressed the capacity of LUAD cells to migrate. Furthermore, the results of cell cycle test displayed that inhibiting circPRKCI could induce the arrest of LUAD cell cycle in the G1 phase. It was discovered through bioinformatics websites that miR-219a-5p had binding sites to circPRKCI 3'UTR, and the results of Dual-Luciferase reporter gene assay revealed that circPRKCI was able to bind to miR-219a-5p. It was uncovered by the qRT-PCR assay results that miR-219a-5p was lowly expressed in LUAD tissues, and its relative expression had an inverse relation with that of circPRKCI according to the Pearson correlation analysis. In addition, it was shown in the results of reverse experiment that miR-219a-5p could regulate the influence of circPRKCI on the malignant phenotype of LUAD. It was found by means of bioinformatics websites that calcium/calmodulin dependent protein kinase ID (CAMK1D) was a downstream target gene of miR-219a-5p and could the two conjugated with each other based on the results of Dual-Luciferase reporter gene assay. Moreover, qRT-PCR assay findings illustrated that CAMK1D was evidently highly expressed in LUAD tissues, and the results of Pearson correlation analysis revealed that CAMK1D expression exhibited a negative association with that of miR-219a-5p and a positive correlation with that of circPRKCI. CONCLUSIONS: CircPRKCI is significantly highly expressed in LUAD, and the highly expressed circPRKCI is capable of facilitating LUAD cell migration, proliferation and cycle. CircPRKCI may regulate the malignant phenotype of LUAD via the miR-219a-5p/CAMK1D axis.

Linc00601 upregulation promotes hepatocellular carcinoma development by activating MAPK signaling pathway.

OBJECTIVE: To study the expression of linc00601 in hepatocellular carcinoma (HCC) tissues and cells, and to study the biological function and downstream mechanism of linc00601 in HCC using in vitro experiments. PATIENTS AND METHODS: The expression of linc00601 in HCC was predicted via bioinformatics, and the expression of linc00601 in HCC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). After interference with the expression of linc00601, the interference efficiency was determined using qRT-PCR, and the changes in HCC cell proliferation, cycle distribution, and apoptosis were determined through Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Finally, the expressions of molecular markers in downstream signaling pathway were determined through Western blotting. RESULTS: It was found via bioinformatics that the expression of linc00601 was upregulated in HCC. The results of qRT-PCR revealed that the expression of linc00601 was upregulated in 36 cases of HCC tissues compared with that in para-carcinoma tissues, and it was also upregulated in HCC cells. According to the results of CCK-8 assay, HCC cell proliferation was inhibited after interference with the expression of linc00601. In the si-linc00601 group, the apoptosis rate rose, and the cell cycle was arrested at the G1/G0 phase compared with those in the si-NC group. The results of Western blotting revealed that after the knockdown of linc00601 in HCC cells, the expressions of molecular markers (p-P38, p-ERK) in the downstream mitogen-activated protein kinase (MAPK) signaling pathway were downregulated. CONCLUSIONS: Linc00601 is upregulated in HCC, which promotes the development of HCC via activating the MAPK signaling pathway.

Assessment of the effect of the Aquamantys® system on local recurrence after hepatectomy for hepatocellular carcinoma through propensity score matching.

OBJECTIVE: To assess the effect of the intraoperative application of the Aquamantys® system to treat the hepatic resection margin on local and overall recurrence of HCC. METHODS: We retrospectively analyzed 101 patients admitted from November 2016 to June 2018 who underwent hepatectomy using the Aquamantys® as hemostatic device, who were matched with 101 patients (control group) using conventional hemostatic devices through PSM. Univariate and multivariate analyses of recurrence-free survival (RFS) and local recurrence-free survival (LRFS) were performed using the Cox proportional hazard model. RESULTS: There were no significant differences in baseline data and surgical procedures between the two groups. The Aquamantys® group showed less blood loss (P = 0.005) and a lower blood transfusion rate (P = 0.036), while the incidences of postoperative complications of the two groups showed no difference (P = 0.266). OS rates of the Aquamantys® group and the control group were 82.6% and 84.2%, respectively (P = 0. 446), and RFS rates were 65.5% and 58.2%, respectively (P = 0.153), with no significant differences. The Aquamantys® group and the control group had two cases and 11 cases of local recurrence, respectively, with LRFS rates of 98% and 87.9%, respectively, in the follow-up period, corresponding to a significant difference (P = 0.011). Multivariate analysis showed that microvascular invasion (MVI), tumor diameter > 5 cm, and the control group were independent risk factors for LRFS. CONCLUSION: Our results indicate that application of the Aquamantys® system in hepatectomy can reduce local recurrence, but it can neither reduce overall recurrence nor improve OS.

LncRNA 00152 promotes the development of hepatocellular carcinoma by activating JAK2/STAT3 pathway.

OBJECTIVE: The aim of this study was to explore the regulatory role of lncRNA 00152 and JAK2/STAT3 pathway in the pathogenesis of hepatocellular carcinoma (HCC), and to investigate the possible underlying mechanism. PATIENTS AND METHODS: Expression levels of lncRNA 00152 in HCC tissues, matched para-cancerous tissues and normal liver tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), respectively. The correlation between lncRNA 00152 expression and pathological characteristics of HCC patients was analyzed. Meanwhile, the expression level of lncRNA 00152 in HCC cell lines was detected by qRT-PCR. After knockdown or overexpression of lncRNA 00152 in MHCC97 or HB611 cells, the proliferative ability and cell cycle were detected by EdU assay and flow cytometry, respectively. Also, Western blot was conducted to detect the protein expression levels of JAK2 and STAT3 in MHCC97 and HB611 cells. RESULTS: The expression of lncRNA 00152 in HCC tissues was significantly higher than that of matched para-cancerous tissues and normal liver tissues. LncRNA 00152 expression was positively correlated with tumor stage and tumor size, whereas negatively correlated with the overall survival of HCC patients. High expression of lncRNA 00152 might be a potential hallmark for the diagnosis of HCC, with the AUC of 0.8425. Similarly, lncRNA 00152 was highly expressed in HCC cell lines when compared with that of normal liver cells. Knockdown of lncRNA 00152 in MHCC97 cells remarkably decreased the proliferative ability and arrested cell cycle. Overexpression of lncRNA 00152 in HB611 cells significantly promoted cell proliferation and cell cycle. Furthermore, Western blot results showed that lncRNA 00152 knockdown upregulated the protein expression levels of JAK2 and STAT3 in HCC cells. CONCLUSIONS: High expression of lncRNA 00152 promotes the development of HCC by activating the JAK2/STAT3 pathway.

Production of medically useful nitric monoxide using AC arc discharge.

Inhaled nitric monoxide (iNO) is increasingly used as a medical treatment for acute respiratory distress syndrome. A course of the existing nitric monoxide (NO) therapy with gas cylinders could cost up to approximately $15,000 for an average of 30.2 h. Moreover, a gas cylinder containing a mixture of N2 and NO may potentially leak NO. The objective of this study is to develop an efficient and cost-effective on-site iNO generation system. In the present setup, NO was generated by using dry air or mixed oxygen/nitrogen (O2/N2) and an AC power source with an output power level of 5-30 W at atmospheric pressure. The simultaneously produced NO2 was eliminated with an ammonium sulfite ((NH4)2SO3) solution. The effects of the O2/N2 ratio, gas flow rate, discharge gap distance, output energy density and electrode structure on NOx concentration and the NO/NO2 ratio are reported. The concentrations of NO and NO2 reached 62 ppm and 3 ppm, respectively, after absorption and dilution at a gas flow rate of 6 L/min. With the present setup, the AC arc discharge produced NOx at a stable concentration for at least 6 h using dry air.

Use of pH as fuzzy control parameter for nitrification under different alkalinity in SBR process.

In order to achieve fuzzy control of nitrification in a Sequencing Batch Reactor (SBR) brewery wastewater was used as the substrate. The effect of alkalinity on pH variation during nitrification was systematically studied, at the same time the variations of DO and ORP were investigated. Alkalinity and pH of the wastewater were adjusted by adding sodium bicarbonate at five levels and sodium hydroxide at two levels. Unadjusted wastewater was also studied. According to the results, variation of pH could be divided into rising type and descending type. When bicarbonate alkalinity was deficient or sufficient, the descending type happened. If alkalinity was deficient, the pH decreasing rate got slower when nitrification nearly stopped; if alkalinity was sufficient, at the end of nitrification pH turned from decrease to increase. This was the most common situation and pH could be used to control the end of nitrification. When alkalinity was excessive, the rising type happened, pH was increasing at nearly a constant rate during and after nitrification and could not be used to control the nitrification time, but if the aeration rate was moderate DO could be used to control the nitrification time. This situation seldom happened. Therefore the variation of pH could not only be used to control the nitrification time but also to judge whether the alkalinity was enough or not. On the basis of this, the fuzzy controller of nitrification in SBR was constructed. When discussing the influence of pH on nitrification rate the composition and concentration of alkalinity must be considered or else the results may be incomprehensive. And to some extent the influence of alkalinity on nitrification rate was more important than pH.

Use pH and ORP as fuzzy control parameters of denitrification in SBR process.

In order to achieve fuzzy control of denitrification in a Sequencing Batch Reactor (SBR) brewery wastewater was used as the substrate. The effects of brewery wastewater, sodium acetate, methanol and andogenous carbon source on the relationships between pH, ORP and denitrification were investigated. Also different quantities of brewery wastewater were examined. All the results indicated that the nitrate apex and nitrate knee occurred in the pH and ORP profiles at the end of denitrification. And when carbon was the limiting factor, through comparing the different increasing rate of pH whether the carbon was enough or not could be known, and when the carbon should be added again could be decided. On the basis of this, the fuzzy controller for denitrification in SBR was constructed, and the on-line fuzzy control experiments comparing three methods of carbon addition were carried out. The results showed that continuous carbon addition at a low rate might be the best method, it could not only give higher denitrification rate but also reduce the re-aeration time as much as possible. It appears promising to use pH and ORP as fuzzy control parameters to control the denitrification time and the addition of carbon.

[The presence of the lymphatic stomata in ovarian bursa of guinea pig and the ultrastructure of ovarian bursa].

In order to explore its structure and speciality of species, the ovary bursa of guinea pig was studied by using dissecting microscopy and scanning electron microscopy. In this paper, the lymphatic stomata on both internal and external layer of the ovary bursa was frist reported in guinea pig. The results suggested that the stomata in bursa not only provided a pathway to connect the bursal cavity with the lymphatic vessels of bursa and the peritoneal cavity, but also may be involved in the reproduction and local immunity of the ovary. The stomata may play an important role in physiological function of the ovary. At the same time, the structural differences were identified in ovary bursa between guinea pig and golden hamster.