Inhibition of autophagy augments chemotherapy in human salivary adenoid cystic carcinoma.

Although cisplatin (DDP)-based adjuvant chemotherapy is widely used in the treatment of salivary adenoid cystic carcinoma (SACC), SACCs have developed resistance to cisplatin, resulting in chemotherapy failure. Autophagy serves as a critical adaptive response, which was increased in tumor cells in chemotherapy. However, the function of autophagy is not clear in SACC. In this study, apoptosis induced by DDP in SACC high metastatic cell line (ACC-M) was revealed using MTT assay, flow cytometry, and caspase-3 immunoblotting. The autophagy activation induced by DDP treatment was measured by transmission electron microscopy, green fluorescent protein-light chain 3 plasmid transfection LC3 immunoblotting and p62 immunoblotting. 3-methyladenine (3-MA) or small interference RNA targeting beclin 1 (beclin 1 siRNA) inhibited autophagy and significantly enhanced DDP-induced apoptosis. ACC-M xenografts in nude mice further verified the synergistic effect of DDP and 3-MA. In conclusion, autophagy activation was caused to protect cancer cells from DDP-induced apoptosis and autophagy inhibition could be a promising strategy for adjuvant chemotherapy in SACC.

Lymphoepithelial carcinoma of the oral cavity and pharynx: a SEER population-based cohort study.

OBJECTIVES: Lymphoepithelial carcinoma (LEC) of the oral cavity and pharynx is a rare cancer, with poorly understood clinicopathological characteristics and prognosis. Only a few case reports or small case series have been reported, so the characteristics and survival of patients with this disease remains unclear. The present study aimed to describe the clinicopathological characteristics and determine the factors associated with survival of this uncommon cancer. METHODS: A population-based study was carried out to investigate clinical characteristics and prognosis of LEC of the oral cavity and pharynx using the data from Surveillance, Epidemiology and End Results (SEER) database. Log-Rank test and Cox regression analysis were performed to determine the prognostic factors, and a prognostic nomogram was further constructed. The propensity-matched analysis was conducted to compare the survival of nasopharyngeal LEC and non-nasopharyngeal LEC patients. RESULTS: Totally, 1025 patients were identified, including 769 nasopharyngeal LEC patients and 256 non-nasopharyngeal LEC patients. The median OS of all patients was 232.0 months (95% CI 169.0-258.0). The 1-, 5-, 10- and 20-year survival rates were 92.9%, 72.9%, 59.3%, and 46.8%, respectively. Surgery significantly prolonedg the survival of LEC patients (P<0.01, mOS: 190 m vs. 255 m). Radiotherapy, as well as radiotherapy after surgery, prolonged the mOS (P<0.01 for both). The survival analysis demonstrated that old age (>60 years), lymph node (N3) and distant metastases were independent factors for poor survival, whereas radiotherapy and surgery were independent factors for favorable survival. The prognostic nomogram was established base on these five independent prognostic factors (C-index = 0.70; 95% CI 0.66-0.74). In addition, no significant difference in survival time between nasopharyngeal LEC and non-nasopharyngeal LEC patients were observed. CONCLUSIONS: LEC of the oral cavity and pharynx is a rare disease, and old age, lymph node and distant metastases, surgery and radiotherapy were significantly associated with prognosis. The prognostic nomogram could be used to make individual predictions of OS.

Survival and differentiation of neuroepithelial stem cells following transplantation into the lateral ventricle of rats.

Neuroepithelial stem cells (NEPs) demonstrate a high potential for self-renewal and differentiation during embryonic development. To explore the survival and differentiation of NEPs in vivo, we isolated NEPs from green fluorescence protein (GFP) transgenic embryos and transplanted into the lateral ventricle of rats. In vitro culture, NEPs proliferated into neurospheres and differentiated into both neurons and glia. When transplanted into the lateral ventricle of rats, these GFP positive NEPs (GFP+ NEPs) survived and attached to the wall of ventricle. Moreover, grafted cells differentiated into neuron-specific enolase (NSE) positive neurons and glial fibrillary acidic protein (GFAP) positive astrocytes and migrated into the host brain. Thus, our results indicate that NEPs can survive and differentiate into neurons and astrocytes in the lateral ventricle following transplantation.

Yolk sac tumor of upper lip: A case report.

Yolk sac tumors (YSTs) are a type of malignant germ cell tumor that usually grow in the gonads. They are difficult to recognize at other sites outside the gonads, and no case has been reported involving the upper lip. The present study reported the case of a 13-month-old girl exhibiting an isolated YST occurring in the upper lip. The histology and elevation of α-fetoprotein were typical for a YST. The patient was cured following effective chemotherapy and surgery resection. After 36 months of follow-up, there was no sign of recurrence or metastasis. A total of 20 cases of primary YSTs of the head and neck extracranial region since 1997 were reviewed. The present study aims to inform the scientific community of the clinical and pathologic features of this patient.

[Correlation between periodontal disease and coronary atherosclerotic heart disease].

OBJECTIVE: To investigate the correlation between coronary atherosclerotic heart disease (CAD) and periodontal disease (PD). METHOD: Forty-five patients with CAD (CAD group) and 40 patients without CAD (control group) were compared with their pathological changes of periodontal tissues and inflammatory markers [high sensitive C reactive protein (hsCRP), interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha)]. RESULTS: Univariate analysis showed that the prevalence of PD was 84.44% in CAD group and 22.50% in control group (P < 0.01). The levels of hsCRP, IL-1beta, and TNF-alpha were (5.75 +/- 1.26) mg/L, (10.32 +/- 2.96) ng/L, and (9.17 +/- 2.14) ng/L in CAD group and (1.13 +/- 0.73) mg/ L, (2.87 +/- 1.45) ng/L, and (5.84 +/- 1.96) ng/L in control group (P < 0.01). Gingival index and plaque index were statistically different between two both groups (P < 0.01). Logistic regression analysis showed that in addition to pulse pressure and low density lipoprotein cholesterol, periodontal disease index was a higher risk factor of CAD. Its relative risk was 1.217 (95% CI was 1.120-1.805, P < 0.05). CONCLUSION: PD can cause CAD. The improvement of public oral health plays an important role in the prevention and treatment of CAD.

Evidence for anaerobic ammonium oxidation process in freshwater sediments of aquaculture ponds.

The anaerobic ammonium oxidation (anammox) process, which can simultaneously remove ammonium and nitrite, both toxic to aquatic animals, can be very important to the aquaculture industry. Here, the presence and activity of anammox bacteria in the sediments of four different freshwater aquaculture ponds were investigated by using Illumina-based 16S rRNA gene sequencing, quantitative PCR assays and (15)N stable isotope measurements. Different genera of anammox bacteria were detected in the examined pond sediments, including Candidatus Brocadia, Candidatus Kuenenia and Candidatus Anammoxoglobus, with Candidatus Brocadia being the dominant anammox genus. Quantitative PCR of hydrazine synthase genes showed that the abundance of anammox bacteria ranged from 5.6 × 10(4) to 2.1 × 10(5) copies g(-1) sediment in the examined ponds. The potential anammox rates ranged between 3.7 and 19.4 nmol N2 g(-1) sediment day(-1), and the potential denitrification rates varied from 107.1 to 300.3 nmol N2 g(-1) sediment day(-1). The anammox process contributed 1.2-15.3% to sediment dinitrogen gas production, while the remainder would be due to denitrification. It is estimated that a total loss of 2.1-10.9 g N m(-2) per year could be attributed to the anammox process in the examined ponds, suggesting that this process could contribute to nitrogen removal in freshwater aquaculture ponds.

Expression of BNIP3 and its correlations to hypoxia-induced autophagy and clinicopathological features in salivary adenoid cystic carcinoma.

BACKGROUND: The expression of Bcl-2/adenovirus E1B 19 kDa-interacting protein3 (BNIP3) has been explored in many human malignancies, but not in adenoid cystic carcinoma (ACC). OBJECTIVE: This study investigated the clinical significance of expression of BNIP3 in ACC tissues and cells and elucidated its correlations to hypoxia-induced autophagy. METHODS: Immunohistochemical and immunofluorescence staining were used to explore BNIP3, HIF-1α and LC3 expression. RESULTS: BNIP3 was positively expressed in 41 cases (63.1%), and was significantly correlated with histological grade (P= 0.001). HIF-1α was positively expressed in 52 cases (80.0%) and was significantly correlated with TNM stage (P= 0.023) and histological grade (P= 0.024). LC3 was positively expressed in 37 cases (56.9%) and was significantly correlated with TNM stage (P= 0.019). The expression of BNIP3 was correlated with HIF-1α expression (P= 0.011). The overall survival in the negative BNIP3 expression group tended to be better than in the positive BNIP3 expression (P= 0.011). In vitro experiment, BNIP3 immunofluorescence staining was detected in cells treated with CoCl2 (for hypoxic condition). CONCLUSIONS: The data indicated that BNIP3 plays a vital role in the tumorigenesis of adenoid cystic carcinoma and could be a new target for gene therapy of adenoid cystic carcinoma.

[Impact of Phosphogypsum Wastes on the Wheat Growth and CO2 Emissions and Evaluation of Economic-environmental Benefit].

Phosphogypsum is a phosphorus chemical waste which has not been managed and reused well, resultantly, causing environmental pollution and land-occupation. Phosphogypsum wastes were used as a soil amendment to assess the effect on wheat growth, yield and CO2 emissions from winter wheat fields. Its economic and environmental benefits were analyzed at the same time. The results showed that wheat yield was increased by 37.71% in the treatment of phosphogypsum of 2 100 kg x hm(-2). Compared with the control treatment, throughout the wheat growing season, CO2 emission was accumulatively reduced by 3% in the treatment of phosphogypsum waste of 1050 kg x hm(-2), while reduced by 8% , 10% , and 6% during the jointing stage, heading date and filling period of wheat, respectively; while CO2 emission was accumulatively reduced by 7% in the treatment of phosphogypsum waste of 2 100 kg x hm(-2) throughout the wheat growing season, as reduced by 11% , 4% , and 12% during the reviving wintering stage, heading date and filling period of wheat, respectively. It was better for CO2 emission reduction in the treatment of a larger amount of phosphogypsum waste. In the case of application of phosphogypsum waste residue within a certain range, the emission intensity of CO2 ( CO2 emissions of per unit of fresh weight or CO2 emissions of per unit of yield) , spike length, fresh weight and yield showed a significantly negative correlation--the longer the ear length, the greater fresh weight and yield and the lower the CO2 emissions intensity. As to the carbon trading, phosphogypsum utilization was of high economic and environmental benefits. Compared with the control, the ratio of input to output changed from 1: 8.3 to 1: 10.7, which in the same situation of investment the output could be increased by 28.92% ; phosphogypsum as a greenhouse gas reducing agent in the wheat field, it could decrease the cost and increase the environmental benefit totally about 290 yuan per unit of ton. The results demonstrated phosphogypsum wastes could obviously decrease the CO2 emission from field soil and had a great potential to control agricultural greenhouse gases. Hopefully it has an important application perspective for the low-carbon, ecological and sustainable agricultural development.

Expression of autophagy and ER stress-related proteins in primary salivary adenoid cystic carcinoma.

Autophagy is the endogenous cellular pathway that facilitates cellular survival by maintaining energy homeostasis and macromolecular synthesis during cellular stress and nutrient deprivation. Endoplasmic reticulum (ER) stress is the process in which disruption of these physiological functions leads to an accumulation of unfolded proteins and induces the unfolded protein response (UPR). ER stress and autophagy are involved in human cancer. We investigated the expression of autophagic proteins (LC3 and beclin 1) and ER stress-related protein (GRP78) in head and neck adenoid cystic carcinoma tissue. Tissue samples from 79 cases of head and neck adenoid cystic carcinoma tissue were utilized for immunohistochemistry. LC3 expression was significantly correlated with lymph node involvement (P=.016) and TNM (P=.021). Beclin 1 expression was significantly correlated with the histological growth pattern (P=.002), the histological grade (P=.000), and longer survival (P=.000). GRP78 expression was significantly correlated with the histological growth pattern (P=.019), the histological grade (P=.019), and longer survival (P=.001). LC3 expression was positively correlated with beclin 1 expression (P=.000); LC3 and beclin 1 expressions were positively correlated with GRP78 expression respectively (P=.035) (P=.008). Our study describes the expression of LC3, beclin 1, and GRP78 in adenoid cystic carcinoma and its relationship with clinicopathologic factors and overall survival. These results suggest that LC3, beclin 1, and GRP78 may play an important role in the tumorigenesis of adenoid cystic carcinoma, and that beclin 1 and GRP78 may serve as new prognostic indicators for the outcome of patients with adenoid cystic carcinoma.

Expression and regulation of amphiregulin in Gsα-mutated human bone marrow stromal cells of fibrous dysplasia of mandible.

OBJECTIVES: Fibrous dysplasia (FD) is a focal bone lesion composed primarily of immature bone marrow stromal cells along with spicules of immature woven bone. However, cellular differentiation and proliferation in mutant human bone marrow stromal cells (hBMSCs) of FD have not been fully elucidated. Therefore, the aim of this study was to investigate the occurrence of G(s)α mutation at the Arg(201) codon in hBMSCs and human trabecular bone cells (hTBCs, osteoblast-like cells). In addition, we evaluated the gene expression and protein secretion of amphiregulin from hBMSCs and hTBCs and assessed the biologic activity and possible mechanism of amphiregulin in our system. STUDY DESIGN: Mutant hBMSCs from FD patients and hTBCs from disease-free bone specimens of the same patient were successfully cultured. We studied the G(s)α mutations at the Arg(201) codon by means of polymerase chain reaction (PCR)-restriction fragment length polymorphism. Gene expression and protein secretion of amphiregulin in hBMSCs and hTBCs was confirmed by reverse-transcription (RT) PCR and Western blotting analysis. The modulation proliferation and possible mechanism of the exogenous addition of amphiregulin and epidermal growth factor receptor tyrosine kinase inhibitor (AG-1478) were assessed by using Wst-1 assays. RESULTS: The G(s)α mutations in clonal adherent mutant hBMSCs and hTBCs were successfully identified. We identified amphiregulin to be highly expressed in hBMSCs compared with hTBCs. The growth of hBMSCs was stimulated by the exogenous addition of amphiregulin and inhibited by AG-1478. CONCLUSIONS: The G(s)α-mutant hBMSCs were successfully identified, and amphiregulin may play a significant role in the proliferation of hBMSCs.

[Experimental studies of human adipose tissue-derived stromal cells transfected with ad-hBMP-2 gene].

OBJECTIVE: To investigate the possibility of adipose-derived stromal cells (ADSCs) transfeced by adenovirus containing human bone morphogenetic protein-2 (Ad-hBMP-2) gene and their osteogenic potential. METHODS: ADSCs were obtained from inguinal fat tissue of 4 weeks old SD rats. After exposure to adenovirus containing green fluorescent protein(Ad-GFP), fluorescent microscope was used to observe gene transfection effect once 12 hours. After transfected with Ad-hBMP-2, cytochemistry, immmucytochemistry and Western blot were used to examine the expression of alkaline phosphatase (ALP), osteocalcin (OC) and hBMP-2. RESULTS: After exposed to Ad-GFP 12 hours, 52% ADSCs were observed being transfected and 48 hours later reached 95%. The double number time belonged after transfecting with Ad-hBMP-2, and cytochemistry, immucytochemistry and Western blot examines indicated positive results of ALP, OC, hBMP-2 after 48 hours. CONCLUSION: Adipose tissue contains abundant ADSCs which could be transfected as gene vectors by adenovirus, ADSCs transfected with Ad-hBMP-2 can convert to ostoeblasts, and can act as a kind of seed cells for osteo-tissue engineering.

[Electromyography study on complete bilateral cleft lip after reparation.].

PURPOSE: This study was undertaken to evaluate the motion effect of complete bilateral cleft lips objectively using electromyography. METHODS: 45 patients of complete bilateral cleft lips after operations several years which included 20 patients repaired using orbicularis oris muscle restoring functional method and 25 patients using straight line suture method were examined with electromyography (EMG) in prolabium, right and left lateral lips respectively. The most potential in every area of each patient at the maximal lip contraction position was measured and the values of two different method groups were subject to statistical analysis. RESULTS: The prolabium and lateral lips of both groups had no electric activity at posture position. At maximal contraction position, the mean potential of prolabium of orbicularis oris muscle restoring method group had no difference with the lateral lips. But the mean potential of prolabium of straight line suture method group was lower than that of the lateral lips significantly,also,it was lower than the potential of prolabium in the muscle functional reparation group. CONCLUSIONS: The reparation method of restoring orbicularis oris muscle to complete bilateral cleft lips was better than the straight line suture method in the motion effect. These findings suggest the superiority and necessity of functional muscle reparation for complete bilateral cleft lips.