[Expression profile of circular RNA in inflammatory response in human bronchial epithelial cells induced by carbon black nanoparticles].

Objective: To explore the toxic effect of carbon black nanoparticles on human bronchial epithelial cells, and identify the differentially expressed circular RNA based on the full transcriptome high-throughput sequencing, so as to provide evidence for the development of biomarkers exposed to carbon black nanoparticles and their application on epigenetic toxicology. Methods: In June 2020, 16 HBE cells were treated with carbon black nanoparticles at concentrations of 20, 40 and 80 µg/ml, and 16 HBE cells without any intervention were used as the control group. The cytotoxicity of carbon black nanoparticles was detected by CCK8 and LDH experiments. Real-time quantitative fluorescent PCR (qRT-PCR) and ELISA were used to detect the changes of interleukin-6 (IL-6) and interleukin-8 (IL-6, IL-8) mRNA and protein levels of carbon black nanoparticles with concentration gradient after 72 h exposure. Western blot analysis was conducted to detect the expression levels of toll-like receptor 4 (TLR4), phosphorylated nuclear factor-κB (P-NF-κB), apoptosis-related speckled protein (ASC) and Caspase-1 associated with nuclear factor-κB. According to high-throughput sequencing results, differentially expressed Circrnas were screened and identified by qRT-PCR, and those with stable differentially expressed circrnas and the strongest association with the NF-κB pathway were selected for ring performance identification. Results: After being exposed to carbon black nanoparticles for 72 h, the activity of 16HBE cells decreased significantly (P<0.05), and the release of lactate dehydrogenase increased significantly (P<0.05). Compared with control group, mRNA expression levels of IL-6 and IL-8, protein levels of IL-6 and IL-8 were increased, and protein levels of TLR4, p-NF-κB, ASC and Caspase-1 were significantly up-regulated in 16 HBE cells of different concentrations, with statistical significance (P<0.05). Compared with the control group, a total of 492 differentially expressed circular Rnas (|log2 FC|>1) were detected. Among the 5 differentially expressed (P<0.05) circular Rnas, circ_002642 was selected as the object of subsequent research on circular Rnas, affter 72 hours of exposure to 80 µg/ml CBNPs, 16HBE cells showed signlficantly higher expression of circ_002642 (P<0.05) . Conclusion: Carbon black nanoparticles can induce differentially expressed circular RNAs associated with inflammatory response in human bronchial epithelial cells.

[Prevention of closure point recanalization after uncut Roux-en-Y anastomosis for radical resection of distal gastric cancer].

Uncut Roux-en-Y anastomosis is widely used in gastrointestinal reconstruction procedure after radical gastrectomy for distal gastric cancer. However, the proximal jejunal closure point recanalization of the input loop is an important complication of postoperative patients with prolonged time, resulting in pancreatic juice or bile reflux, which can lead to inflammatory lesions of the remnant stomach or esophagus. Poor selection of the location of the closure point during anastomosis causes a large amount of food deposited in the blind loop to be pushed and impacted, resulting in loosened threads or failed U-shaped staples, which may cause recanalization complications. Most scholars believe that the shortening of the jejunal tube closure point to the optimal position of 2 to 3 cm from the residual gastrojejunostomy can significantly reduce food retention, decrease the pressure of the closure point and the incidence of recanalization. At present, the application of new anastomotic techniques and materials such as four-row and six-row U-shaped staples and 7# wire ligation under laparoscopy can prevent the occurrence of recanalization of the closure point. Uncut Roux-en-Y anastomosis is safe and has few complications, and is expected to become one of the best ways of digestive tract reconstruction.

The potential mechanism of INHBC and CSF1R in diabetic nephropathy.

OBJECTIVE: The aim of this study was to research the potential mechanism of INHBC and CSF1R in diabetic nephropathy. MATERIALS AND METHODS: 30 SD rats were selected and randomly divided into Con group, Sham group, and DN group. In the DN group, intraperitoneal injection of the streptozotocin-citrate solution was conducted to construct the DN model. In the Sham group, intraperitoneal injection of equal citrate solution was conducted. The Con group did not do anything. After successful modeling, blood glucose, insulin, biochemical indexes, and levels of inflammatory cytokines in blood samples were detected. The expression levels of INHBC, CSF1R, apoptosis-related proteins and IGF-1 were detected by Western blot. MRNA expression levels of INHBC, CSF1R, IGF-1 and inflammatory cytokines were detected by qPCR. RESULTS: Compared with the Con group, the expression levels of blood glucose, insulin, biochemical indexes, INHBC, CSF1R, IGF-1, IL-6, TNF-α and Bcl2 increased in the DN group, while the expression levels of IL-10, Caspase 3, Caspase 9, and Bax decreased. INHBC mRNA was positively correlated with IGF-1 mRNA. CSF1R was negatively correlated with Caspase 3, Caspase 9, Bax, and IL-10, and positively correlated with IL-6, TNF-α, and Bcl2. CONCLUSIONS: NHBC and CSF1R induced the secretion of IL-6 and TNF-α, inhibited the production of IL-10, inhibited apoptosis of cells, and promoted the proliferation of renal cells during DN disease. Therefore, INHBC and CSF1R can be used as target objects of DN treatment strategies.

MicroRNA-214 functions as an oncogene in human osteosarcoma by targeting TRAF3.

OBJECTIVE: Osteosarcoma is a malignant bone tumor with high incidence. The prognosis of osteosarcoma is very poor when it is diagnosed with metastasis. Numerous studies have demonstrated that aberrant expressions of microRNAs are involved in cancer initiation and development. However, the potential role of miR-214 in osteosarcoma remains largely unrevealed. The current study investigated the relationship between the miR-214 and TNF receptor-associated factor 3 (TRAF3) in osteosarcoma tissues and cell lines. We also aimed to evaluate the potential roles of miR-214 on the occurrence and metastasis in osteosarcoma and verify its effect on the regulation of TRAF3. PATIENTS AND METHODS: The miR-214 expression and TRAF3 expression in osteosarcoma tissue samples and cell line were measured using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Followed by transfection assays, transwell assay was conducted to detect the migration and invasion abilities of osteosarcoma cells. Subsequently, Western blotting and luciferase reporter assay were performed in osteosarcoma cells to confirm the target of miR-214. RESULTS: The results showed that miR-214 expression levels were significantly increased not only in osteosarcoma tissues but also in osteosarcoma cell lines as compared with adjacent normal tissues and matched cell lines, respectively. On the contrary, the TRAF3 expression levels in osteosarcoma tissues and cell lines were frequently decreased compared to the control group. Moreover, TRAF3 was identified as a direct target of miR-214 and the inverse relationship between them was also observed in osteosarcoma tissues. Additionally, we found that miR-214 restoration could significantly promote osteosarcoma cell invasion and migration via targeting TRAF3. CONCLUSIONS: MicroRNA-214 functioned as an oncogene in osteosarcoma via targeting TRAF3, which may provide new insights into osteosarcoma prevention and treatment.

[Effect of APP on prognosis in patients with chronic rhinosinusitis after endoscopic sinus surgery].

Objective:To investigate interventional effect of APP on prognosis in patients with chronic rhinosinusitis after endoscopic sinus surgery. Method:One hundred and forty-four chronic rhinosinusitis patients in our hospital were divided into observation group and control group randomly; observation group had 71 patients, and control group had 73 patients. The control group was treated with standard discharge instruction. In addition to standard discharge instruction, observation group was followed up and directed by using of APP. Two groups' VAS scores and Lund-Kennedy scores were compared in admission time, and 6 months after discharge. Result:There was no significant difference in the Lund-Kennedy scores and VAS scores between the two groups in admission time. However, 6 months after discharge, Lund-Kennedy scores and VAS scores in observation group were obviously lower than control group(P< 0.05). Conclusion:The use of healthy APP can significantly promote patients with chronic rhinosinusitis recovery and re-visit after endoscopic sinus surgery.

MiR-216b is involved in pathogenesis and progression of hepatocellular carcinoma through HBx-miR-216b-IGF2BP2 signaling pathway.

This study aims to investigate the expression status of miRNA-216b in familial hepatocellular carcinoma (HCC) and the correlation between miRNA-216b expression and pathogenesis, as well as the progression of HCC. The expression profile of miRNAs in plasma of peripheral blood between HCC patients with HCC family history and healthy volunteers without HCC family history was determined by microarray. Using real-time quantitative PCR to detect the expression in paired tissues from 150 patients with HCC, miR-216b was selected as its expression value in HCC patients was significantly lower compared with healthy volunteers. Next, miR-216b expression and the clinicopathological features of HCC were evaluated. The effect of miR-216b expression on tumor cells was investigated by regulating miR-216b expression in SMMC-7721 and HepG2 in vitro and in vivo. Finally, we explored mRNA targets of miR-216b. In 150 HCC, 37 (75%) tumors showed reduced miR-216b expression comparing with their adjacent liver tissues. The decreased expression of miR-216b was significantly correlated with tumor volume (P=0.044), HBV infection (P=0.026), HBV DNA quantitative (P=0.001) and vascular invasion (P=0.032). The 5-year disease-free survival and overall rates after liver resection in low expression and high expression groups of miR-216b are 62% and 54%, 25% and 20%, respectively. MiR-216b overexpression inhibited cell proliferation, migration and invasion, and miR-216b inhibition did the opposite. The expression of hepatitis B virus x protein (HBx) has tight correlation with downregulation of miR-216b. Furthermore, miR-216b downregulated the expression of insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) and exerted its tumor-suppressor function through inhibition of protein kinase B and extracellular signal-regulated kinase signaling downstream of IGF2. MiR-216b inhibits cell proliferation, migration and invasion of HCC by regulating IGF2BP2 and it is regulated by HBx.

Associations between clinical characteristics and oncogene expression in patients with non-small cell lung cancer.

More than 40 oncogenes associated with non-small cell lung cancer (NSCLC) have been identified with varied gene expression. The correlations between specific clinical characteristics and oncogene expression in NSCLC patients were examined. From October 2011 to September 2012, a total of 60 patients with NSCLC (male:female, 34:24; mean age, 59.5 ± 10.6 years; age range, 31-81 years) were diagnosed and evaluated for treatment with radical resection at a single facility. Eligible patients exhibiting tumor node metastasis (TNM) stage I-III NSCLC confirmed by post-surgical pathology were included. mRNA expression was detected by branched DNA-liquidchip technology (bDNA-LCT) and mutations were detected at EGFR exons 18, 19, 20, and 21, KRAS exons 2 and 3, BRAF and PIK3CA exons 9 and 20. Correlations between gene expression at mutations and clinical characteristics of gender, age, histological type, degree of differentiation, smoking status, immunohistochemical (IHC) evaluation of TTF-1, TNM staging, and discrete age ("nage") were examined. Significant associations were observed between IHC staining for TTF-1 and histological type (P = 0.00001) and with BRAC1, TYMS, RRM1, and TUBB3 expression (P = 0.0187, 0.0051, 0.024, and 0.0238, respectively). Significant cross-correlations were observed between TYMS, BRAC1, TOP2A, STMN1, TUBB3, and RRM1 expression (P < 0.05), but not between EGFR exon 21, KRAS exon 2, and PIK3CA exon 9 expression and any other mutation expression (P > 0.05). Relationships between clinical characteristics and oncogene expression in NSCLC, particularly those of TTF-1 level and smoking status, may be useful indicators of prognosis and development of anti-cancer drug resistance.

Plasmid-based short hairpin RNA against connective tissue growth factor attenuated monocrotaline-induced pulmonary vascular remodeling in rats.

Pulmonary hypertension is a life-threatening medical condition, and a growing body of evidence shows that the expression of connective tissue growth factor (CTGF) is significantly associated with its pathogenesis, making it an attractive therapeutic target. Our earlier work revealed that plasmid-based CTGF-specific short hairpin RNA (shRNA) could attenuate pulmonary artery smooth muscle cell (PASMC) proliferation and pulmonary vascular remodeling in rats exposed to cigarette smoke. In this study, we explored the therapeutic role of this shRNA plasmid in the treatment of monocrotaline-induced pulmonary vascular remodeling in rats, and demonstrated that the upregulation of CTGF in PASMCs following a single injection of monocrotaline could be attenuated by administration of the shRNA. Accordingly, this shRNA was found to repress monocrotaline-induced pulmonary vascular remodeling, as evidenced by its ability to reduce the percentage of muscularized vessels and the wall thickness of pulmonary vessels. We concluded that plasmid-based shRNA against CTGF attenuated pulmonary vascular remodeling in monocrotaline-treated rats. CTGF might be a potential target for the treatment of pulmonary vascular remodeling and pulmonary hypertension.

Cerebral hyperaemia after isoflurane anaesthesia for craniotomy of patients with supratentorial brain tumour.

BACKGROUND: Few studies look into cerebral blood flow (CBF) changes during emergence from general anaesthesia for craniotomy. The purpose of this study was to assess CBF changes during emergence from general anaesthesia for craniotomy, through monitoring blood oxygen saturation of jugular vein bulb (SjvO2 ) and transcranial Doppler (TCD). METHODS: We enrolled 30 patients undergoing selective craniotomy (group C) for supratentorial brain tumour resection and 30 patients undergoing selective abdominal surgery (group A). Mean velocity of middle cerebral artery (Vmca), mean arterial pressure (MAP), SjvO2 (only measured in group C), and arterial CO2 partial pressure were measured before anaesthesia, at tracheal extubation, and 30, 60, 90, 120 min after extubation. RESULTS: Vmca of the same side of tumour was significantly higher than contralateral Vmca before anaesthesia and at all times after extubation in group C. The ipsilateral Vmca increased significantly (95.7 ± 16.9 cm/s vs. 63.7 ± 6.7 cm/s, P < 0.01) at extubation in group C, then declined but still above baseline significantly in the first 2 h after extubation. While Vmca of the right side changed only slightly (63.6 ± 7.7 cm/s vs. 61.8 ± 8.1 cm/s, P < 0.01) but significantly at extubation in group A. SjvO2 increased significantly (81.4% ± 7.4% vs. 60.9% ± 3.7%, P < 0.01) at extubation in group C, and remained above baseline significantly for 2 h. There was no significant correlation between Vmca and MAP at any time. CONCLUSIONS: Cerebral hyperaemia occurs after supratentorial brain tumour resection surgery. The hyperaemia is more pronounced on the same side as the tumour.

Should we lower the dose of iron when treating anaemia in pregnancy? A randomized dose-response trial.

BACKGROUND/OBJECTIVES: To compare the efficacy and side effects of low-dose vs high-dose iron supplements to correct anaemia in pregnancy. SUBJECTS/METHODS: One hundred and eighty women with anaemia (haemoglobin <110 g l(-1)) in mid-pregnancy. The women were randomly allocated to 20; 40 or 80 mg of iron daily for 8 weeks from mid-pregnancy. RESULTS: One hundred and seventy-nine (99%) women completed the trial. At the end of treatment, there was a clear dose-response of increasing mean haemoglobin concentration with iron dose (111+/-13 g l(-1) at 20 mg per day, 114+/-11 g l(-1) at 40 mg per day and 119+/-12 g l(-1) at 80 mg per day, P=0.006). However, the incidence of anaemia did not differ statistically between groups. Compared with women in the 80 mg iron group, the odds ratio of anaemia was 1.9 (95% CI: 0.8, 4.3, P=0.130) and 1.1 (95% CI: 0.5, 2.6, P=0.827), respectively, for women in the 20 mg iron group and the 40 mg iron group. The incidence of gastrointestinal side effects was significantly lower for women in the 20 mg iron group compared with women in the 80 mg iron group; the odds ratio was 0.4 (95% CI: 0.2, 0.8, P=0.014) for nausea, 0.3 (95% CI: 0.2, 0.7, P=0.005) for stomach pain and 0.4 (95% CI: 0.2, 0.9, P=0.023) for vomiting. CONCLUSIONS: Low-dose iron supplements may be effective at treating anaemia in pregnancy with less gastrointestinal side effects compared with high-dose supplements.

[Clinical and experimental study on gastrosia convalescens in treating chronic atrophic gastritis].

Gastrosia convalescens (GC) is a new drug for treating the Spleen-Yin Deficiency syndrome of chronic atrophic gastritis (CAG). Animal experiment has shown that GC has the function of promoting the regeneration of atrophic glands, and that of anti-inflammation, reinforcing, strengthening the stomach etc. It has the effect on the intestinal electric activity in dogs, mainly exciting in action, which was superior than that of domperidoni (P < 0.05). The result of clinical study has demonstrated that the marked effective rate of GC was 78.32%, CAG reversed into chronic superficial gastritis (CSG) in 30.0% of patients (60/200 cases), CAG into chronic superficial atrophic gastritis (CSAC) in 39.0% (78/200 cases) and CSAG into CSG in 39.02% (48/123 cases). The disappearance rate of metaplasia of intestinal epithelium and atypical hyperplasia were 39.1% and 38.14% respectively. There was a very significant difference in comparing with control group (P < 0.01).

Electron microscopic observations of tegumental damage in adult Schistosoma japonicum after in vivo treatment with levo-praziquantel.

The in vivo effects of a single dose of levo-praziquantel, 75 mg/kg in PEG 400, on the tegumental surface of adult S. japonicum were compared with the effects of a single dose (150 mg/kg) of the mixed isomer preparation, using scanning and transmission electron microscope. Worms were recovered from mice at 10 min, 30 min, 1 hr, 4 hr, 12 hr, 24 hr and 48 hr after treatment. After 10 min exposure to either compound, the tegumental folds and sensory organelles were swollen and the tegument vacuolated. After 12-24 hr, the surface was eroded and exfoliated with exposure of intrategumental and/or subtegumental tissues and attachment of leukocytes to the denuded areas. Vehicle controls were normal throughout the time period examined. These studies demonstrate that the levo isomer of praziquantel causes acute structural damage to the tegument similar to that seen with the mixed isomer preparation.

[Antitumor activity of monoclonal antibody MI2 against immunosuppressive acidic protein in vitro].

In vitro antitumor activity of monoclonal antibody MI2 that was made by our laboratory to direct against immunosuppressive acidic protein (IAP) was observed with MTT assay for cytotoxicity. The results showed that the growth of human gastric cancer cell line SGC 7901 was inhibited significantly (P < 0.01) when MI2 was added at a concentration of 7.81 mg/L or higher. The inhibition activity of MI2 appeared to be dose dependent. Increased cytotoxicity (up to 206.3%) of LAK cells against SGC7901 could be remarkably (P < 0.01) induced by addition of MI2 at a concentration of 1.95 mg/L, so the ratio of effector to target was 10:1. The enhancing effect of MI2 on LAK cell activity was also dose dependent. The antitumor activity of MI2 was not associated with human complements.

[Establishment of immuno-PCR technique for the detection of tumor associated antigen MG7-Ag on the gastric cancer cell line].

The gastric cancer associated antigen McAb MG7-Ag was detected by means of a newly established method, termed immuno-PCR. A McAb-recombinant DNA chimeric molecule was made which possesses bispecific binding affinity for antigen that had been immobilized on microtiter wells and the segment of the attached DNA was amplified by PCR. The antigen of gastric cancer cell line KATO III was monitored by this method. Analysis of PCR products by agarose gel electrophoresis after staining with ethidium bromide allowed as few as 20 cells to be detected readily and reproducibly. Immuno-PCR showed a 10(4) enhancement in detection sensitivity compared with ELISA assay. When the same numbers of cells (2 x 10(6)/ml) were immobilized and then the serial diluted chimeric molecule was added, 3.8 x 10(-14) moles and 3.0 x 10(-11) moles were needed to give positive results with the immuno-PCR and ELISA assay, respectively. Therefore, immuno-PCR could give an enormous amplification capability with good specificity, and has a sensitivity much higher than any existing techniques for antigen detection.

Age-dependent change in the inhibitory effect of an adenosine agonist on hippocampal acetylcholine release in rats.

To investigate the possibility that age-dependent deficits in acetylcholine (ACh) release are precipitated by the alteration of endogenous purinergic activities, the effects of (-)N6-phenylisopropyladenosine (PIA), an adenosine agonist, in modulating K+ (25 mM)-induced [3H]ACh release from the hippocampal slices of young (3-6 months old) and old rats (26-30 months old) were examined. In young rats, PIA (0.1-10 microM) caused a dose-related inhibition of [3H]ACh release from the hippocampal slices and a significant reduction in [3H]ACh release was observed in the presence of 1 microM PIA. In old rats, a similar pattern of PIA suppression of K(+)-induced [3H]ACh release was observed; however, a 10-fold higher concentration of PIA (10 microM) was required to elicit a significant inhibition. This age-dependent reduction in responsiveness to PIA may be due to an enhanced endogenous adenosine activity in aged rats leading to downregulation of the adenosine receptors. This notion is supported by the finding that both the adenosine concentration and activity of 5'-nucleotidase, an enzyme partially governing adenosine synthesis, were increased in the hippocampus of old rats as compared to their younger counterparts.