Circ_0067934 correlates with poor prognosis and promotes laryngeal squamous cell cancer progression by sponging miR-1324.

OBJECTIVE: As a kind of non-coding RNA, circular RNA (circRNA) plays a regulatory role in many tumors. Our intention was to investigate the clinical significance, biological function, and molecular regulation mechanisms of circ_0067934 in laryngeal squamous cell cancer (LSCC) progression. PATIENTS AND METHODS: The expression levels of circ_0067934 in LSCC tumor samples and cells were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The associations of circ_0067934 expression with clinicopathological features and overall survival in LSCC patients were statistically analyzed. The biological function of circ_0067934 in LSCC cells was analyzed by CCK-8, colony formation, EdU, and transwell assays. Dual-Luciferase reporter assay was conducted to verify that circ_0067934 may sponge miR-1324 to regulate proliferation and migration of LSCC. RESULTS: Circ_0067934 expression was remarkably upregulated in LSCC tissues and cells. High level of circ_0067934 was significantly related to tumor size, lymph node status, and distant metastasis of LSCC. Furthermore, highly expressed circ_0067934 could result in notably worse survival. In vitro experiments showed that down-regulation of circ_0067934 inhibited proliferation and migration of LSCC cells. Mechanism analysis revealed that circ_0067934 regulated LSCC development by sponging miR-1324. CONCLUSIONS: Circ_0067934 may function as an oncogene in LSCC, which provided a feasible prognostic biomarker and therapeutic target for LSCC.

MiR-16 inhibits hepatocellular carcinoma progression by targeting FEAT through NF-κB signaling pathway.

OBJECTIVE: MicroRNA-16 (miR-16) expression has been proved to take part in the initiation and development of several cancers, including hepatocellular carcinoma (HCC). However, its role and its molecular mechanism in HCC cells remain unclear. Our study aimed to elucidate miR-16 probable role and potential mechanism in HCC cells. PATIENTS AND METHODS: MiR-16 expression in HCC was measured by Real Time-Polymerase Chain Reaction (RT-PCR). MiR-16 mimic or inhibitor was applied to increase or decrease miR-16 expression in Huh7 cells separately. The cell viability was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). The invaded cells and migrated cells were detected by the transwell assay. The epithelial-mesenchymal transition (EMT) and the nuclear factor-κB (NF-κB) were performed using Western blot. The tumor growth was measured via xenograft tumor formation assay. Moreover, bioinformatical methods and luciferase reporter assay were carried out to confirm the miR-16 target gene. RESULTS: MiR-16 expression was downregulated in HCC tissues and cells. Furthermore, the increasing miR-16 expression was suppressed, whereas the decreasing miR-16 expression promoted cell proliferation, invasion, and migration in Huh7 cells. Moreover, miR-16 targeted FEAT in regulating HCC progression. FEAT was associated with a poor prognosis of HCC patients. Especially, miR-16 suppressed EMT and NF-κB pathway in HCC and inhibited the tumor growth in vivo. CONCLUSIONS: We stated that miR-16 suppressed HCC cell progression by targeting FEAT and inhibiting EMT and NF-κB pathway. MiR-16 may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.

[Prevalence and distribution of hypertension in population aged 15 years and over in Guizhou province].

Objective: To understand the prevalence and distribution of hypertension in population aged 15 years and over in Guizhou province and provide evidence for the prevention and management of hypertension. Methods: Face to face interviews using national standard questionnaire were conducted among the study subjects selected in Guizhou through multi-stage random sampling. Blood pressure measurement for them was done with Omron HEM-1300 professional portable blood pressure monitor. SPSS 19.0 software was used for statistical analysis. The ratio was compared by the χ(2) test. The influencing factors of hypertension was analyzed by multivariate logistic regression analysis. Results: A total of 13 480 participants were investigated, including 5 509 (40.8%) men and 7 971 (59.2%) women; 6 558 (48.6%) urban residents and 6 922 (51.4%) rural residents. Among the subjects surveyed, 3 232 (23.9%) were smokers, 2 412 (17.9%) were alcoholic and 4 859 (36.0%) were obese or overweight. A total of 3 937 (29.2%) hypertension patients were found. The prevalence of hypertension was 29.2%. The standardized prevalence of hypertension were 18.97% (compared with national population composition) and 21.16% (compared with Guizhou province population composition), respectively. The hypertension prevalence in men and women were 29.8% and 28.8%, respectively. The hypertension prevalence in rural population (35.8%) was higher than that in urban population (22.2%). The difference was statistically significant (P<0.001). The hypertension prevalence in people aged 65 years and over was 56.2%. The prevalence of hypertension were 34.3% and 27.6% in smokers and non-smokers, 39.2% and 27.0% in alcoholic and non-alcoholic and 40.7% and 22.7% in obese or overweight group and normal or less weight group, respectively. There were significant statistical differences in prevalence of hypertension among the population in urban area and rural area, with different age, education levels, smoking status, drinking status and BMI (P<0.001). Conclusions: The prevalence of hypertension in Guizhou was at a high level. The hypertension prevalence in rural area was higher than that in urban area. Hypertension prevalence increased significantly with age. The prevalence of hypertension was negatively associated with the education level of the people. Older age, living in rural area, smoking, drinking, obesity were the risk factors for hypertension.

[Multiple-factor analysis of serum allergen distribution of patients with allergic rhinitis and level of main allergen IgE in Shenyang area].

Objective:To discuss multiple-factor analysis of serum allergen distribution of patients with allergic rhinitis and level of main allergen IgE in Shenyang area, and to provide a scientific basis for the prevention of allergic rhinitis in this area.Method:Serum IgE was detected in 749 cases［501 cases of male (66.8%), and 248 cases of female (33.2%)］, with allergic rhinitis.The age range was from 3 to 65 years old, and they were divided into 5 groups based on age. A questionnaire survey was conducted to analyze the distribution of serum allergens and to carry out a multiple-factor analysis of level of the main allergen IgE in patients.Result:The primary allergen was house dust mite/dust mite in each age group, and the differences in the positive rate of elm, mold, cat/dog fur scurf, cockroach and ragweed among different groups were statistically significant（P<0.01 or P<0.05）.The ingested allergen sequencing for all age groups: the distribution of 7-14 years old group and 15-35 years old group were consistent with that of the whole, among the under 6 years old group, mango and pineapple was ranked 1st,beef and mutton was ranked 2nd,the occurrence rate of shrimp and crab rose to the 3rd place, among the 36-60 years old group, mango, pineapple and milk was ranked 1st, egg was ranked 2nd, and beef and mutton ranked 3rd, the differences in positive rate of mango and pineapple, beef and mutton, crab and nut among all groups were statistically significant(P<0.01 or P<0.05).The IgE level of cockroach was impacted by the allergic history, home cultivation of flowers and plants and animal domestication. The IgE level of mold was impacted by sex, allergy history, home rearing of pet and furniture updates. The IgE level of wormwood was impacted by allergy history and asthma history. The IgE level of peanut was impacted by age and allergy history. The IgE level of egg was impacted by history of food and drink allergy, home cultivation of flowers and plants and home rearing of pet. The IgE level of crab was impacted by the allergy history and home rearing of pet.Conclusion: The primary inhaled allergen in all groups is house dust mite/dust mite, and the ingested allergen varies in each group, which has provided a reference basis for prevention of ingested allergy.The varying influence factors for IgE level of primary allergens could be used to prevent the patients from contacting the allergen,and each risk factor has become a focus of prevention and control for patients, offering a major help to the prevention and treatment of allergic rhinitis.

Effect of FGF2 on the activity of SPRYs/DUSP6/ERK signaling pathway in endometrial glandular epithelial cells of endometriosis.

OBJECTIVE: We aimed at exploring the positive feedback loop in eutopic and ectopic endometrial glandular epithelial cells (EuECs and EECs) in endometriosis. MATERIALS AND METHODS: Normal epithelial cells (NECs), EuECs and EECs were treated with fibroblast growth factor (FGF)2, FGF2 neutralizing antibody, mitogen-activated protein kinases (MAPKs) inhibitors U0126 and PD98059. FGF2 protein level was detected by enzyme-linked immunosorbent assay (ELISA). The expressions of FGF2, FGF receptor 1 (FGFR1), extracellular signal-regulated kinase (ERK)1/2/pERK1/2 and Sproutys (SPRYs) (Sprouty1, Sprouty2, Sprouty4) and dual specificity phosphatase 6 (DUSP6) were detected by Western blot. The mRNA levels of FGF2, FGFR1 (FGF receptor 1), SPRYs (Sprouty1, Sprouty2, Sprouty4) and DUSP6 mRNA were detected by RT-PCR. RESULTS: Among treatment groups, the content of FGF2 in EuECs and EECs was significantly higher than that in NECs (p < 0.05). The mRNA and protein levels of FGF2, FGFR1, SPRYs (Sprouty1, Sprouty2, Sprouty4) and DUSP6 in EuECs and EECs were increased after adding FGF2 (p < 0.05), but decreased after adding FGF2 neutralizing antibody, no significant change was found in NECs (p > 0.05). The inhibitory effect of PD9805 on NECs was not significantly different from that of U0126 (p > 0.05); however, the inhibitory effects of PD9805 on EuECs and EECs were significantly lower than those of U0126 (p< 0.05). CONCLUSIONS: The positive feedback loop existed in EuECs and EECs, but maybe not in NECs. The results may provide the guideline to treat endometriosis patients.

Transient expressions of doppel and its structural analog prionDelta32-121 in SH-SY5Y cells caused cytotoxicity possibly by triggering similar apoptosis pathway.

Doppel (Dpl) is a recently identified prion (PrP)-like protein due to the structural and biochemical similarities, however, its natural function and pathogenic role in neurodegenerative diseases remains unclear. To investigate the possible pathogenic pathway of Dpl and its structural analog for cell apoptosis, mammalian expressing recombinant plasmids containing human PRND gene encoding the full-length Dpl and a truncated human PRNP gene deleting the sequences encoding the peptide from aa 32 to 121 (PrPDelta32-121) were generated. MTT assays showed the cell viabilities of the human neuroblastoma cell line SH-SY5Y receiving Dpl and PrPDelta32-121 expressing plasmids were remarkably lower. Obvious apoptosis phenomena were observed to be associated with the cells transient expressing Dpl and PrPDelta32-121, including reduced mitochondrial transmembrane potential (psim), decreased pro-caspase-3 quantity, more numbers of annexin V- and annexin V/PI-double-stained cells and depressed Bcl-2 level. Moreover, we also found that the Dpl- and PrPDelta32-121-induced cytotoxicities and relevant apoptotic events in SH-SY5Y cells could be fully antagonized by co-expression of the human full-length PrP. These data highly indicate that cytotoxicity induced by the expression of Dpl and truncated PrP in neural derived cells are closely related with the apoptosis process, probably triggering the mitochondrial pathway. It also implies that the cell-benefit activity of the full-length PrP may result from its anti-apoptosis capacity.

Immunization of mice with plasmids coexpressing HPV16 E5 and the novel oncogene hWAPL.

The novel human oncogene hWAPL is associated with uterine cervical cancer. The HPV16 E5 oncoprotein could induce genomic instability in normal human cells. However, the mechanism of E5 interaction with hWAPL still awaits definition. In our present studies, the eukaryotic expression plasmids, pcDNA3-hWAPL and pcDNA3-hWAPL-E5 were constructed and carried out to vaccinate mice directly. The result that indicated the polyclonal antibody titer in immunized mice sera was increased by enzyme-linked immunosorbent assay. In addition, the proliferative responses of immunized mice spleen cells showed the optical densities values in vaccinated group remarkably higher than that in the control group. In conclusion, the recombinant plasmids could induce strong humoral and cellular immune response and exhibited great potential as therapeutic targets in the treatment of cervical cancer. However, the result didn't show significant difference in group with coexpression of HPV16 E5-hWAPL and group with only hWAPL expression. Consistent with these observations, we demonstrated that HPV16 E5 was not the optimal factor to cooperate with hWAPL in gene therapy.

The relationship between HPV16 and expression of CD44v6, nm23H1 in esophageal squamous cell carcinoma.

The esophageal squamous cell carcinoma (ESCC) has high incidence in Shaanxi Province of China. More and more researches indicated that human papillomavirus type 16 (HPV16) might play an important role in carcinogenesis of ESCC but the relationship between HPV16 and CD44v6, nm23H1 has not been elucidated. HPV16 was detected by amplifying HPV16 E6 gene through polymerase chain reaction (PCR) method and the expression of CD44v6, nm23H1 in 40 ESCCs and fifteen normal esophageal mucosa (NEM) from Shaanxi Province was examined by Streptavidin-Peroxidase (SP) method using monoclonal antibody specific to CD44v6 and nm23H1. The positive rates of HPV16 E6 gene, CD44v6 and nm23H1 were 60% (24/40), 65% (26/40) and 45% (18/40) respectively in ESCCs and 26.67% (4/15), 33.33% (5/15) and 86.67% (13/15) respectively in NEMs. There exited statistical difference for HPV16, CD44v6 and nm23H1 between NEMs and ESCCs respectively (p < 0.05). The relationship between HPV16 and the expression of CD44v6 in ESCCs was statistical significance (P = 0.021), but no significant correlation was found between HPV16 and the expression of nm23H1 (P = 0.436) in ESCCs. The infection rate of HPV16 had no statistical difference in all pathological features we observed, but the expression rates of CD44v6 and nm23H1 had statistical correlation with invasion (p = 0.001, 0.013) and lymph nodes metastasis (p = 0.014, 0.002) respectively. In different histology grade of ESSCs, the relationship between HPV16 and CD44v6 was statistical significance in grade I (p = 0.044) but was not in grade II (p = 0.165) and grade III (p = 0.658), however as to the expression of nm23H1 there exited no statistical significance in all histology grades of ESCC (p > 0.05). The expression rates of CD44v6 and nm23H1 were statistically different between grade I and II (p = 0.004, 0.016) respectively and between grade I and grade III (p = 0.014, 0.020), but not statistically different between grade II and III (p = 0.792, 0.943) respectively. Our data firstly suggested that there existed the statistical relationship between the infection of HPV16 and the expression of CD44v6 in ESCCs and that HPV16 may be involved in invasion and metastasis of ESCC.

Long-term propofol infusion and airway management in a patient with Goldenhar's syndrome.

A 2-year-old patient of Goldenhar's syndrome received an operation for corneal transplantation. Difficult endotracheal intubation from the congenital anomaly was treated with laryngeal mask airway and pediatric fiberoptic laryngoscope. Long-term propofol infusion (> 10 h) for anaesthetic maintenance in this small child was used with rapid and smooth recovery.

[Congenital coronary artery-cardiac chamber fistula: diagnosis and surgical treatment].

From Nov. 1976 to Dec. 1989, 9 patients with congenital coronary artery-cardial chamber fistula were operated upon in our hospital. In this series right coronary artery-right ventricle fistula was found in 3 patients, right coronary artery-left ventricle fistula in 2, left coronary artery-right ventricle fistula in 2, left circumflex artery-right atrium fistula in 1, and bilateral coronary artery-cardiac chamber fistula in 1. Cardiopulmonary bypass was performed in 8. The fistula was closed by interrupted mattress sutures outside the cardiac chamber plus ligation of terminal portion of the coronary artery near the fistula in 5 patients, and by mattress sutures inside the dilated coronary artery or cardiac chamber in other 4. There was no operative death in the 8 patients with single fistula. One patient with bilateral coronary artery fistula died from hypoxemia postoperatively. Late complications or recurrent fistulas were not found in the follow-up period from 3 months to 12 years. The diagnosis, operative indications and technique of closing fistula were discussed.

[Gene diagnosis of familial erythroleukemia at the early stage of leukomogenesis].

Southern blot hybridization was performed with probe v-erbB + A in 4 subjects. Two of them were patients with similar erythroid alterations of bone marrow: one of them was diagnosed as erythroleukemia (EL). The other two were members of a same family (husband and wife). The husband had normal erythroid cellularity of bone marrow but the wife and their 7 living sons and daughters had erythroid hypercellularity. In the second generation of this family 4 hematological patients (1 with AML and 3 with CAA) were found in the past decade. Rearrangement of cerbB and c-erbA genes was found in 3 of the 4 subjects (2 with EL and the wife mentioned above). They had 4 new hybridization bands besides 3 germlines, but the husband had only 3 germlines. The result suggests that using Southern blot hybridization technique with probe v-erbB + A, the rearrangement of c-erbB/c-erbA genes might be a diagnostic indicator of erythroleukemia at early stage of leukomogenesis in familial and sporadic patients. The etiological significance of the rearrangement of c-erbB/c-erbA in leukomogenesis was discussed.

Hydrolysis of nicotinamide-adenine dinucleotide by purified renal brush-border membranes. Mechanism of NAD+ inhibition of brush-border membrane phosphate-transport activity.

Purified rat renal brush-border membrane vesicles possess a heat-labile enzyme activity which hydrolyses NAD+. A reciprocal relationship exists between the disappearance of NAD+ and the appearance of adenosine; 2 mol of Pi are liberated from each mol of NAD+ incubated with brush-border membrane vesicles. Freezing and thawing brush-border membrane vesicles does not enhance the initial rate of NAD+ hydrolysis. Preincubation of brush-border membrane vesicles with NAD+ results in inhibition of Na+-dependent Pi-transport activity, whereas Na+-dependent glucose transport is not affected. EDTA, which prevents the release of Pi from NAD+ and which itself has no direct effect on brush-border membrane Pi transport, reverses the NAD+ inhibition of Na+-dependent Pi transport. These results suggest that it is the Pi liberated from NAD+ and not NAD+ itself that inhibits Na+-dependent Pi transport.