[Molecular Mechanism of Busulfan Inhibiting the Malignant Biological Characteristics of Multiple Myeloma Cells].

OBJECTIVE: To investigate the effect of Busulfan on the malignant biological characteristics of multiple myeloma cells and explore the molecular mechanism. METHODS: Multiple myeloma RPMI8226 cells were treated with Busulfan at different concentrations. Cell proliferation activity was detected by MMT assay, and cell apoptosis was detected by flow cytometry with Annexin V/PI double staining. Real-time quantitative PCR was used to detect the mRNA expression of apoptotic regulatory molecules Bax、Bcl-2 and Wnt3a/ß-catenin pathway, and Western blot was used to detect the expression changes of Bax, Bcl-2 and Wnt3a/ß-catenin pathway protein. RESULTS: Busulfan inhibited cell proliferation and induced apoptosis of myeloma RPMI8226 cells (P<0.05). After treatment with Busulfan at different concentrations for 48 h, the expression of anti-apoptotic protein Bcl-2 was decreased, the expression of pro-apoptotic protein Bax was up-regulated, and the activation of Wnt3a/ß-catenin signaling pathway was inhibited to induce programmed death of RPMI8226 cells (P<0.05). CONCLUSION: Busulfan can inhibit the malignant biological characteristics of myeloma RPMI 8226 cells, and its mechanism may be related to regulating the expression of Bcl-2 family proteins and inhibiting the activation of Wnt3a/ß-catenin signaling pathway.

Construction of a circRNA-Mediated ceRNA Network Reveals Novel Biomarkers for Aortic Dissection.

Background: Aortic dissection (AD) is a rare and lethal disorder with its genetic basis remains largely unknown. Many studies have confirmed that circRNAs play important roles in various physiological and pathological processes. However, the roles of circRNAs in AD are still unclear and need further investigation. The present study aimed to elucidate the underlying molecular mechanisms of circRNAs regulation in AD based on the circRNA-associated competing endogenous RNA (ceRNA) network. Methods: Expression profiles of circRNAs (GSE97745), miRNAs (GSE92427), and mRNAs (GSE52093) were downloaded from Gene Expression Omnibus (GEO) databases, and the differentially expressed RNAs (DERNAs) were subsequently identified by bioinformatics analysis. CircRNA-miRNA-mRNA ceRNA network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to predict the potential functions of circRNA-associated ceRNA network. RNA was isolated from human arterial blood samples after which qRT-PCR was performed to confirm the DERNAs. Results: We identified 14 (5 up-regulated and 9 down-regulated) differentially expressed circRNAs (DEcircRNAs), 17 (8 up-regulated and 9 down-regulated) differentially expressed miRNAs (DEmiRNAs) and 527 (297 up-regulated and 230 down-regulated) differentially expressed mRNAs (DEmRNAs) (adjusted P-value <0.05 and | log2FC | > 1.0). KEGG pathway analysis indicated that DEmRNAs were related to focal adhesion and extracellular matrix receptor interaction signaling pathways. Simultaneously, the present study constructed a ceRNA network based on 1 circRNAs (hsa_circRNA_082317), 1 miRNAs (hsa-miR-149-3p) and 10 mRNAs (MLEC, ENTPD7, SLC16A3, SLC7A8, TBC1D16, PAQR4, MAPK13, PIK3R2, ITGA5, SERPINA1). qRT-PCR demonstrated that hsa_circRNA_082317 and ITGA5 were significantly up-regulated, and hsa-miR-149-3p was dramatically down-regulated in AD (n = 3). Conclusion: This is the first study to demonstrate the circRNA-associated ceRNA network is altered in AD, implying that circRNAs may play important roles in regulating the onset and progression and thus may serve as potential biomarkers for the diagnosis and treatment of AD.

Lipopolysaccharide induces pyroptosis through regulation of autophagy in cardiomyocytes.

BACKGROUND: Autophagy, a stress response in eukaryotic cells, is closely related to cardiogenic diseases. Pyroptosis, a newly discovered way of programmed cell death, also plays an important role in cardiovascular disease. However, the role and relationship of autophagy and pyroptosis in lipopolysaccharide (LPS)-induced inflammatory response of cardiomyocytes were still unclear. METHODS: Western blot was performed to determine the expression of poly ADP-ribosepolmesera-1 (PARP-1), LC3B, NLRP3 and GSDMD in cardiomyocytes after the treatment of LPS. Transfection of si-LC3B, western blot and immunofluorescence (IF) staining were performed to investigate the role of autophagy in LPS-induced pyroptosis. Co-immunoprecipitation (Co-IP) assays and quantitative real-time PCR (qRT-PCR) were conducted to explore whether PARP-1 binding to LC3B and modulating its expression. Transfections of si-PARP-1, western blot and IF were carried out to confirm the role of PARP-1 in the regulation of LPS-induced pyroptosis by autophagy. RESULTS: LPS induces autophagy and pyroptosis in cardiomyocytes, enhanced the level of autophagy and inhibited the level of pyroptosis in the concentration of 4 µg/mL. We further proved that autophagy inhibits LPS-induced pyroptosis in cardiomyocytes. In addition, PARP-1 binding to LC3B and regulate the expression of LC3B. Finally, we proved that knockdown of PARP-1 rescued the inhibition of autophagy on LPS-induced pyroptosis of cardiomyocytes. CONCLUSIONS: LPS induces pyroptosis through regulation of autophagy via PARP-1 at a specific concentration, above which it causes deposition of autophagy flow to promote pyroptosis. Inhibiting LPS-induced pyroptosis could be a promising therapeutic target in treating cardiovascular diseases.

Factors associated with trait anger level of juvenile offenders in Hubei province: A binary logistic regression analysis.

The risk factors of high trait anger of juvenile offenders were explored through questionnaire study in a youth correctional facility of Hubei province, China. A total of 1090 juvenile offenders in Hubei province were investigated by self-compiled social-demographic questionnaire, Childhood Trauma Questionnaire (CTQ), and State-Trait Anger Expression Inventory-II (STAXI-II). The risk factors were analyzed by chi-square tests, correlation analysis, and binary logistic regression analysis with SPSS 19.0. A total of 1082 copies of valid questionnaires were collected. High trait anger group (n=316) was defined as those who scored in the upper 27th percentile of STAXI-II trait anger scale (TAS), and the rest were defined as low trait anger group (n=766). The risk factors associated with high level of trait anger included: childhood emotional abuse, childhood sexual abuse, step family, frequent drug abuse, and frequent internet using (P<0.05 or P<0.01). Birth sequence, number of sibling, ranking in the family, identity of the main care-taker, the education level of care-taker, educational style of care-taker, family income, relationship between parents, social atmosphere of local area, frequent drinking, and frequent smoking did not predict to high level of trait anger (P>0.05). It was suggested that traumatic experience in childhood and unhealthy life style may significantly increase the level of trait anger in adulthood. The risk factors of high trait anger and their effects should be taken into consideration seriously.

Factors Associated with Trait Anger Level of Juvenile Offenders in Hubei Province: A Binary Logistic Regression Analysis / 华中科技大学学报（医学）（英德文版）

The risk factors of high trait anger of juvenile offenders were explored through question naire study in a youth correctional facility of Hubei province,China.A total of 1090 juvenile offenders in Hubei province were investigated by self-compiled social-demographic questionnaire,Childhood Trauma Questionnaire (CTQ),and State-Trait Anger Expression Inventory-Ⅱ (STAXI-Ⅱ).The risk factors were analyzed by chi-square tests,correlation analysis,and binary logistic regression analysis with SPSS 19.0.A total of 1082 copies of valid questionnaires were collected.High trait anger group (n=316) was defined as those who scored in the upper 27th percentile of STAXI-Ⅱ trait anger scale (TAS),and the rest were defined as low trait anger group (n=766).The risk factors associated with high level of trait anger included:childhood emotional abuse,childhood sexual abuse,step family,frequent drug abuse,and frequent internet using (P＜0.05 or P＜0.01).Birth sequence,number of sibling,ranking in the family,identity of the main care-taker,the education level of care-taker,educational style of care-taker,family income,relationship between parents,social atmosphere of local area,frequent drinking,and frequent smoking did not predict to high level of trait anger (P＞0.05).It was suggested that traumatic experience in childhood and unhealthy life style may significantly increase the level of trait anger in adulthood.The risk factors of high trait anger and their effects should be taken into consideration seriously.

Minimally invasive perventricular device closure of ventricular septal defect: a comparative study in 80 patients.

OBJECTIVE: To evaluate the efficacy of minimally invasive perventricular device closure of ventricular septal defect (VSD). METHODS: Between September 2011 and February 2013, we collected 40 patients who underwent perventricular closure via a small lower sternal incision (minimally invasive group), aged 15.5±3.5 years (12 months to 32 years) with a body weight of 24.2±7.5 kg (10.8-58.0 kg). The mean size of VSD was 5.6±0.5 mm (2-14 mm). Another 40 patients were included as the surgical group, receiving the conventional surgical repair of VSD. The device of the minimally invasive group was released under the guidance of transesophageal echocardiography. Success rate, cardiac indicators, and clinical outcomes of the 2 groups were compared. RESULTS: The patients in the surgical group and those in the minimally invasive group showed similar results in success rate (both 97.5%). The procedure time, intensive care unit stay, hospital stay, and postoperative recovery time in the minimally invasive group were significantly shorter than those in the surgical group (58±21 minutes versus 145±26 minutes, 2±1 days versus 8±3 days, 5±1 days versus 16±6 days, 3±1 days versus 90±20 days, all P<0.05). The minimally invasive group had a higher incidence of conduction anomalies (17.5% versus 2.5%, P<0.05). In the follow-up period of 3-12 months, there was no new residual shunt, noticeable aortic regurgitation, significant arrhythmias, or device failure except for new complications in the surgical group. CONCLUSIONS: The success rate of minimally invasive perventricular device closure of VSD under transesophageal echocardiography guidance is similar to that of conventional surgical repair, but the short-term outcomes of the minimally invasive approach is much better. Long-term follow-up is necessary to confirm the effectiveness of this technique.

[Isoflurance-based intravenous and inhalation combined anesthesia versus low-dose-ketamine-based total intravenous anesthesia for valvuloplasty in minipigs].

OBJECTIVE: To compare the effectiveness of anesthesia between Isoflurane-based intravenous and inhalant combined approach and low-dose-ketamine-based total intravenous approach for valvuloplasty in minipigs. METHODS: Twenty four minipigs were given 3-5 mg/kg ketamine intramuscularly and 15-20 mg/kg pentobarbital intravenously for anesthetic induction and intubation. They were then randomly divided into two groups, each with 12 minipigs. In group I (isoflurane), the minipigs received isoflurane 1.0-2.0 minimum alveolar concentration (MAC), fentanyl 20-25 microg/(kg x h), midazolam 0.1-0.2 mg/(kg x h) and pipecuronium 0.10-0.15 mg/(kg x h) for maintenance. In group K (ketamine), the minipigs were given ketamine 3-5 mg/(kg x h), pentobarbital 8-10 mg/(kg x h) and pipecuronium 0. 10-0.15 mg/(kg x h) intravenously. The general peri-operation characteristics were recorded. Hemodynamics, blood gas and respirations were monitored. Anesthetic complications were observed. RESULTS: Two minipigs died from causes other than anesthesia. The minipigs in group I had lower mean aortic pressure (MAP) than those in group K during cardiopulmonary bypass (CPB), but without a statistical significance. The minipigs in group I had significantly lower levels of lactic acid than those in group K after CPB cessation (P < 0.05). The times on analepsia were (21.6 +/- 4.1) min and (67.8 +/- 8.5) min for group I and group K, respectively. The times on ventilator were (281.3 +/- 34.7) min and (330.4 +/- 27.0) min for group I and group K, respectively. The differences were significant (P < 0.05). One minipig in group K was intubated for espiratory depression after surgery. CONCLUSION: The isoflurane-based intravenous and inhalant combined anesthesia was preferable for valvular reparation in minipigs. However, low-dose-ketamine-based total intravenous anesthesia is also a good choice in the circumstance of limited resources.

Immunocytochemical detection of HoxD9 and Pbx1 homeodomain protein expression in Chinese esophageal squamous cell carcinomas.

AIM: To evaluate the expression pattern of two novel oncofetal antigens, the HoxD9 and Pbx1 homeoproteins in esophageal squamous cell carcinomas (ESCCs) to determine what role they would play in the carcinogenesis of ESCC. METHODS: We obtained tissue samples of ESCC from 56 patients who underwent esophagectomy but not preoperative chemotherapy or radiotherapy. The diagnosis of ESCC was established and confirmed by staff pathologists. We used a highly sensitive, indirect, immunocytochemical method to detect HoxD9 and PbX1 proteins. We qualitatively and quantitatively evaluated cells that exhibited and staining using a light microscope. RESULTS: In all observed carcinoma tissue samples, more than 60% of neoplastic cells stained lightly or strongly for HoxD9, and more than 50% of neoplastic cells stained lightly or strongly for Pbx1. CONCLUSION: Our data suggest that HoxD9 and Pbx1 are inappropriately expressed in most human esophageal squamous cell carcinoma. Understanding the role of Hox genes in esophageal epithelial cell carcinogenesis may not only augment early detection but also offer new avenues for treatment of this disease.

[Expression of p53 and vascular endothelial growth factor in esophageal squamous cell carcinoma and their clinical significance].

BACKGROUND & OBJECTIVE: Experimental study proved that the coexpression of p53 and vascular endothelial growth factor (VEGF) play an important role in angiogenesis of tumor. The aim of this study was to investigate the mechanism that p53 participate angiogenesis and the relationship between the expression of p53 and VEGF and clinical pathologic parameters and prognosis of esophageal squamous cell carcinoma (ESCC). METHODS: The expressions of p53 and VEGF in operative samples from 76 ESCC patients were detected by immunohistochemistry. The vascular endothelial cells in tumor tissue were labeled by F VIII factor antibody for counting microvessel density (MVD). RESULTS: The expression rates of p53 and VEGF were 60.5% and 56.5%; total expression rate was 42.1%. The expression rates of p53 and VEGF were strongly associated with distal metastasis and vascular infiltration of ESCC (P < 0.05, P < 0.01). Distal metastasis and vascular infiltration usually occurred in the patients whose mutant p53 and VEGF were both positive expression. The MVDs in p53(+) or VEGF(+) (31.7 +/- 11.5; 33.8 +/- 11.7) were both significantly higher than that in p53(-) or VEGF(-) (22.4 +/- 10.6; 21.2 +/- 9.3, P < 0.05). The MVD reached the maximum in the patients whose p53 and VEGF were both positive. CONCLUSION: Mutant p53 expression is closely associated with the angiogenesis and distal metastasis of ESCC; Expression of p53 and VEGF could be used as an important biological indices for evaluating the malignant degree of ESCC. Combined determination of p53 and VEGF expression has important clinical significance.