Laryngeal contact granuloma after radiotherapy in patients with nasopharyngeal carcinoma: a case series.

Background: Laryngeal contact granuloma (LCG) is a benign hypertrophic lesion and phonatory injury after abnormal vocal behavior is regarded as its major etiology. Patients receiving radiation for non-laryngeal head and neck tumors are troubled by persistent voice impairment. The occurrence of LCG after radiotherapy for nasopharyngeal carcinoma (NPC) in our practice has implored us to re-exam their underlying etiology. We hypothesize that a proportion of LCG results from voice change caused by non-laryngeal head and neck cancer radiotherapy and firstly describe a distinct LCG population originated after radiotherapy for NPC with respect to the clinical profile, presentation, prognosis and response to treatment of patients. Methods: We retrospectively reviewed the laryngoscopic examination and tumor study findings to elucidate the common clinical features of patients who presented with LCG after radiotherapy for NPC. All patients were regularly monitored with telescopic examination until lesions disappeared. Data on age, sex, clinical presentation, telescopic findings, management, latency time of lesion formation, remission time and clinical outcome were reviewed. Results: The medical review identified 27 cases of LCG secondary to radiotherapy for NPC. All lesions had been diagnosed during routine endoscopy following radiation. The interval between radiation onset and endoscopic diagnosis was 3.77 months (range, 0.67-11 months). 20 cases were resolved through simple observation, 4 cases were resolved with the administration of proton pump inhibitors (PPIs), and 3 cases with a poor response to PPI therapy required subsequent surgical resection. The mean remission time in the observation and PPI groups was 4.42 months (range, 0.73-18.9 months) and 5.78 months (range, 2.17-14.63 months), respectively. All patients recovered completely and none experienced recurrence during a mean follow-up of 32.44 months (range, 5.6-71.67 months). Conclusions: Iatrogenic granulomas of vocal process are presenting after radiation for non-laryngeal head and neck cancers. In contrast with spontaneous granulomas, these granulomas can be cured at high remission rates and low recurrence trend without specific intervention. Thus, simple observation may be sufficient for radiation-induced LCG.

Establishment of Immortalized Laryngeal Epithelial Cells Transfected with Bmi1.

Primary laryngeal epithelial cells are essential to exploring the mechanisms of laryngeal and voice disorders; however, they are difficult to study and apply because of their limited life span. The purpose of this study was to develop a stable and reliable in vitro model for the comprehensive study of the pathogenesis of laryngeal and voice diseases. The pLVTHM-Bmi1 plasmid was constructed and used to immortalize primary laryngeal epithelial cells by lentiviral infection. The expressions of Bmi1, human telomerase reverse transcriptase (hTERT), p53, and pRB pathway proteins were detected by western blotting. Functional characteristics of the immortalized cell lines were verified by cell senescence ß-galactosidase staining, 5-ethynyl-2'-deoxyuridine cell proliferation test, and flow cytometry. We successfully introduced Bmi into human subglottic (hSG) cells and human ventricle (hV) cells. Both the human immortalized subglottic Bmi1 (hSG-Bmi1) cell line and the human immortalized ventricle Bmi1 (hV-Bmi1) cell line maintained normal epithelial morphology and divided successfully after more than 20 culture passages. As Bmi1 was overexpressed in these cells, the expression of human telomerase reverse transcriptase (hTERT) and phosphorylated Rb increased while p16 and p21 decreased. Following Bmi1-mediated immortalization, cell senescence decreased significantly, and cell proliferation was accelerated. Tumor formation was not observed for hSG, hV, or hSG-Bmi1, and hV-Bmi1 cells in nude mice. hSG-Bmi1 cells dominated by stratified squamous epithelium and hV-Bmi1 cells dominated by columnar cells were established. The new cell lines lay a foundation for the study of the pathogenic mechanisms of laryngeal and voice diseases.

Attitudes Toward Advance Directives Among Patients and Their Family Members in China.

OBJECTIVES: Chinese people are generally unfamiliar with the concept of advance care planning or advance directives (ACP/ADs), which raises dilemmas in life-support choice and can even affect clinical decision making. To understand and address the issues involved better, we investigated the awareness of ACP/ADs in China, as well as people's attitudes toward medical autonomy and end-of-life care. DESIGN: A multicenter cross-sectional survey, conducted from August 1 to December 31, 2016. SETTING: Twenty-five hospitals located in 15 different provinces throughout mainland China. PARTICIPANTS: Pairs of adult patients without dementia or malignancies, and a family member. MEASUREMENTS: Participants self-filled anonymous questionnaires, and the data collected were analyzed to relate patients' sociodemographic characteristics to their awareness of ACP/ADs and attitudes to health care autonomy and end-of-life care. RESULTS: Among 1084 patients who completed the questionnaire, 415 (38.3%) had heard about ACP/ADs. Having been informed about ACP/ADs, 995 (91.8%) were willing to find out their true health status and decide for themselves; 549 (50.6%) wanted to institute ACP/ADs. Regarding end-of-life care, 473 (43.6%) chose Do Not Resuscitate, and 435 (40.1%) wished to forgo life-support treatment if irreversibly moribund. Patients predominantly (481, 44.4%) chose general hospital as their preferred place to spend their last days of life; only 114 (10.5%) favored a special hospice facility. Patients' main concerns during end-of-life care were symptom control (35.1%), followed by functional maintenance and quality of life (29.8%), and prolonging life (18.9%). More highly educated patients had significantly greater awareness of ACP/ADs than less well educated ones (χ2 = 59.22, P < .001) and were more willing to find out the truth for themselves (χ2 = 58.30, P ≤ .001) and make medical decisions in advance (χ2 = 55.92, P < .001). Younger patients were also more willing than older ones to know the truth (χ2 = 38.23, P = .001) and make medical decisions in advance (χ2 = 18.42, P = .018), and were also more likely to wish to die at home (χ2 = 96.25, P < .001). Only 212 patients' family members (19.6%) wanted life-support treatment for themselves if irreversibly moribund, whereas 592 (54.6%) would want their relative to receive such procedures in the same circumstances; a similar discrepancy was evident for end-of-life invasive treatment (18.3% vs 42.7%). CONCLUSIONS: Awareness about ACP/ADs in China is still low. Providing culturally sensitive knowledge, education, and communication regarding ACP/ADs is a feasible first step to promoting this sociomedical practice.

Rutaecarpine ameliorates hyperlipidemia and hyperglycemia in fat-fed, streptozotocin-treated rats via regulating the IRS-1/PI3K/Akt and AMPK/ACC2 signaling pathways.

AIM: We have shown that rutaecarpine extracted from the dried fruit of Chinese herb Evodia rutaecarpa (Juss) Benth (Wu Zhu Yu) promotes glucose consumption and anti-inflammatory cytokine expression in insulin-resistant primary skeletal muscle cells. In this study we investigated whether rutaecarpine ameliorated the obesity profiles, lipid abnormality, glucose metabolism and insulin resistance in rat model of hyperlipidemia and hyperglycemia. METHODS: Rats fed on a high-fat diet for 8 weeks, followed by injection of streptozotocin (30 mg/kg, ip) to induce hyperlipidemia and hyperglycemia. One week after streptozotocin injection, the fat-fed, streptozotocin-treated rats were orally treated with rutaecarpine (25 mg·kg(-1)·d(-1)) or a positive control drug metformin (250 mg·kg(-1)·d(-1)) for 7 weeks. The body weight, visceral fat, blood lipid profiles and glucose levels, insulin sensitivity were measured. Serum levels of inflammatory cytokines were analyzed. IRS-1 and Akt/PKB phosphorylation, PI3K and NF-κB protein levels in liver tissues were assessed; pathological changes of livers and pancreases were examined. Glucose uptake and AMPK/ACC2 phosphorylation were studied in cultured rat skeletal muscle cells in vitro. RESULTS: Administration of rutaecarpine or metformin significantly decreased obesity, visceral fat accumulation, water consumption, and serum TC, TG and LDL-cholesterol levels in fat-fed, streptozotocin-treated rats. The two drugs also attenuated hyperglycemia and enhanced insulin sensitivity. Moreover, the two drugs significantly decreased NF-κB protein levels in liver tissues and plasma TNF-α, IL-6, CRP and MCP-1 levels, and ameliorated the pathological changes in livers and pancreases. In addition, the two drugs increased PI3K p85 subunit levels and Akt/PKB phosphorylation, but decreased IRS-1 phosphorylation in liver tissues. Treatment of cultured skeletal muscle cells with rutaecarpine (20-180 µmol/L) or metformin (20 µmol/L) promoted the phosphorylation of AMPK and ACC2, and increased glucose uptake. CONCLUSION: Rutaecarpine ameliorates hyperlipidemia and hyperglycemia in fat-fed, streptozotocin-treated rats via regulating IRS-1/PI3K/Akt signaling pathway in liver and AMPK/ACC2 signaling pathway in skeletal muscles.

Proteomic identification of alpha-2-HS-glycoprotein as a plasma biomarker of hypopharyngeal squamous cell carcinoma.

Hypopharyngeal squamous cell carcinoma (HSCC) has very poor prognosis compared with other head and neck squamous cell carcinomas. Late-stage diagnosis of HSCC increases mortality. Therefore, more effective biomarkers for early diagnosis of HSCC are necessary. Unfortunately, appropriate biomarkers for clinical diagnosis and prognosis have not been identified yet. However, recent progresses in quantitative proteomics have offered opportunities to identify plasma proteins as biomarkers for HSCC. In the present study, plasma samples were analyzed by two-dimensional differential gel electrophoresis (2D-DIGE), and differentially expressed proteins were identified by matrix assisted laser desorption ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). A total of 26 proteins representing 12 unique gene products were identified. The up-regulation proteins were alpha-2-HS-glycoprotein (AHSG), complement C4-B, haptoglobin, C-reactive protein, and ceruloplasmin, whereas the down-regulation proteins were serum albumin, angiotensinogen, alpha-1-antichymotrypsin, Ig gamma-3 chain C region, fibrinogen gamma chain, apolipoprotein A-I, and Ig kappa chain C region. Among all the differentially expressed proteins, AHSG was validated by western blot and ELISA. The results were consistent with the data from 2D-DIGE, further suggesting that AHSG may be employed as a potential biomarker for the early diagnosis of HSCC. In summary, this study was the first to use 2D-DIGE and MALDI-TOF/TOF platform to identify the potential plasma biomarkers for HSCC. The plasma AHSG showed great potential for HSCC screening.

MiR-124 suppresses tumor growth and metastasis by targeting Foxq1 in nasopharyngeal carcinoma.

BACKGROUND: The molecular mechanisms underlying dysregulation of microRNAs have been documented in nasopharyngeal carcinoma (NPC). Our previous study demonstrated that plasma miR-124 was down-regulated in NPC using microarray analysis and quantitative PCR validation. Though growing studies showed that down-regulated miR-124 was closely related to tumourigenesis in various types of cancers, the role of miR-124 in NPC remains largely unknown. METHODS: The expression level of miR-124 was evaluated in NPC cell lines and patient specimens using quantitative reverse transcription-PCR (Real-time qPCR). The clinicopathological significance of the resultant data was later analyzed. Then, we explored the role of miR-124 in NPC tumorigenesis by in vitro and in vivo experiments. Homo sapiens forkhead box Q1 (Foxq1) was confirmed as a novel direct target gene of miR-124 by the dual-luciferase assay and western bolt. RESULTS: We found that miR-124 was commonly down-regulated in NPC specimens and NPC cell lines. The expression of miR-124 was inversely correlation with clinical stages and marked on T stages. Then, the ectopic expression of miR-124 dramatically inhibited cell proliferation, colony formation, migration and invasion in vitro, as well as tumor growth and metastasis in vivo. Furthermore, we identified Foxq1 as a novel direct target of miR-124. Functional studies showed that knockdown of Foxq1 inhibited cell growth, migration and invasion, whereas Foxq1 overexpression partially rescued the suppressive effect of miR-124 in NPC. In clinical specimens, Foxq1 was commonly up-regulated in NPC, and the level increased with clinical stages and T stages. Additionally, the level of Foxq1 was inversely correlated with miR-124. CONCLUSIONS: Our results demonstrate that miR-124 functions as a tumor-suppressive microRNA in NPC, and that its suppressive effects are mediated chiefly by repressing Foxq1 expression. MiR-124 could serve as an independent biomarker to identify patients with different clinical characteristics. Therefore, our findings provide valuable clues toward the understanding the of mechanisms of NPC pathogenesis and provide an opportunity to develop new effective clinical therapies in the future.

Levels of brain natriuretic peptide are associated with peripheral arterial disease in subjects with type-2 diabetes mellitus.

BACKGROUND: The effects of brain natriuretic peptide (BNP) on the risk of cardiovascular disease and atherosclerosis have been studied. However, little information is available regarding peripheral arterial disease (PAD), particularly among subjects with type-2 diabetes mellitus (T2DM). The aim of our study was to assess the potential relationship between BNP levels and PAD among T2DM patients. METHODS: The study cohort was 507 T2DM outpatients in which BNP levels were measured. Cross-sectional associations between BNP levels (in tertiles) and PAD were examined. RESULTS: Compared withT2DM patients without PAD, BNP levels were markedly higher in patients with PAD (p = 0.001). Correlation analyses showed that the BNP level was negatively correlated with the ankle-brachial index (r = -0.453, p = 0.033). At a cutoff value of 78.2 pg/ml, the BNP level showed a sensitivity of 71.9%, a specificity of 68.1%, and a positive predictive value of 84.3% for a diagnosis of PAD. The area under the receiver-operating characteristic curve increased significantly if BNP levels were incorporated into a predictive model of the potential risk factors for PAD (0.85 vs 0.81, p = 0.029). CONCLUSIONS: BNP is a potential and promising biomarker for PAD screening in T2DM patients.

MicroRNA-378 functions as an onco-miR in nasopharyngeal carcinoma by repressing TOB2 expression.

Increasing evidence indicates that microRNAs (miRNAs) has been implicated in the progression and metastasis of numerous cancers. In particular, abnormal expression of miR-378 has been observed in various cancers and is associated with cell survival, migration, invasion, angio-genesis and tumor growth. Our previous studies have shown that miR-378 was decreased in nasopharyngeal carcinoma (NPC) plasma and was negatively correlated with NPC progression. However, the tissue expression of miR-378 and its biological function remained unknown in NPC. In this study, we report for the first time that expression level of miR-378 was commonly upregulated in both NPC tissues and NPC cell lines compared to normal healthy nasopharyngeal epithelial samples and human nasopharyngeal epithelial cell lines (NP69), respectively, and was opposite to the reported results in plasma. Functional studies showed that upregulation of miR-378 dramatically promoted cell proliferation, colony formation, migration and invasion in vitro, as well as tumor growth in vivo. Bioinformatics analyses were performed to predict the target genes of miR-378, and the following mechanistic investigations revealed that miR-378 overexpression was able to downregulate the expression of transducer of ERBB2 (TOB2), a potential tumor suppressor, and miR-378 silencing enhanced TOB2 expression. In clinical specimens, TOB2 was widely repressed in tumor tissues accompanied by miR-378 overexpression. Taken together, this study indicates that miR-378 regulates TOB2 and may function as an onco-miR in NPC progression, providing a potential target for gene therapy of NPC.

Positivity of both plasma Epstein-Barr virus DNA and serum Epstein-Barr virus capsid specific immunoglobulin A is a better prognostic biomarker for nasopharyngeal carcinoma.

BACKGROUND: Positivity of plasma Epstein-Barr virus (EBV)-DNA or serum virus capsid antigen-specific IgA (VCA-IgA) is a biomarker for the prognosis of nasopharyngeal carcinoma (NPC). The objective of this study was to determine the value of positivity for plasma EBV-DNA and/or VCA-IgA in predicting the survival of patients with NPC. METHODS: Plasma EBV-DNA and serum VCA-IgA in 506 NPC patients in this retrospective study were detected by quantitative real time polymerase chain reaction and enzyme-linked immunoabsorbent assay, respectively. The value of positivity for EBV-DNA and/or VCA-IgA in predicting the survival of patients with NPC was analyzed. RESULTS: Patients with positivity for both EBV-DNA and VCA-IgA had significantly shorter periods of relapse free survival (RFS) and overall survival (OS) than those with positive single measure or negative for both measures, and patients with positive single measure had significantly shorter periods of RFS and OS than those with negative for both. Multivariate analysis indicated that the positivity for EBV-DNA and/or VCA-IgA were significant risk factors for shorter periods of RFS and OS. CONCLUSION: These data indicated that positivity for both EBV-DNA and VCA-IgA was a better biomarker for the prognosis of patients with NPC. Our findings may provide new references for clinical practice.

[Pathologic changes of the palatopharyngeal muscles in adult patients with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To investigate the pathology of palatopharyngeal muscle obtained from patients with obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: The samples from both groups were studied under HE, nicotinamide adenine dinucleotide-tetrazolium reductase (NADH- TR), modified Gomori trichrome (MGT) and adenosine triphosphatase (ATPase) staining. There were 36 cases of OSAHS who received uvulopalatopharyngoplasty in the experimental group (including 6 mild, 6 moderate and 24 severe cases). There were 6 patients with chronic tonsillitis but without OSAHS as matched control group. Both groups were diagnosed by PSG. RESULTS: Centralized located nuclei and obvious variability of the size of fiber types were observed in both groups. The occurrence rate of the former were 1/6 in control group and 52.8% (19/36) in OSAHS, while the rate of the latter were 4/6 and 58.3% (21/36)respectively. A large number of fibers in both groups (control group 5/6, OSAHS group 28/36) presented an irregularly distributed staining for oxidative activity reaction in NADH stain.Endomysium connective tissue proliferation, a lobular or motheaten appearance, target-like fibers, ragged red fiber (RRF) and muscle necrosis were only observed in OSAHS group.While it was more common in serious OSAHS patients. Dominance of type 1 fibers were observed in matched control group in ATPase stain. Clusters of type 2 fibers or clusters of both type fibers were observed in OSAHS, especially more common in serious OSAHS. There was a predominance of the type 2 fibers in some OSAHS patients. CONCLUSIONS: The observation of HE and special muscular stain identified that palatopharyngeal muscle of OSAHS patients had pathological lesion. The pathological changes included muscular lesion and abnormal distribution of different fiber types, the rate of type 1 fiber which maintained the opening of upper air way decreased.

Serum galectin-3: a risk factor for vascular complications in type 2 diabetes mellitus.

BACKGROUND: Plasma galectin-3, a mediator of fibrogenesis and inflammation, its potential to associate with type 2 diabetes (T2DM) is poorly investigated. Here, we explored its interaction with the serum galectin-3 and vascular complications. METHODS: We conducted a population-based cross-sectional survey in Zhejiang, China involving 165 men and 119 women (age range, 43 - 84 years), investigating the relationship between serum galectin-3 and vascular disease in patients with T2DM. RESULTS: Serum galectin-3 was higher in subjects with T2DM than that in control participants (27.4 vs. 17.6 ng/ml, P < 0.001). Compared with subjects with galectin-3 values in the lowest quartile, those with values in the highest quartile had an increased likelihood of vascular complications (4th quartile odds ratio (OR) 2.52, 95% confidence interval (CI), 1.25 - 4.07). Increased risk of micro- or macrovascular complications correlated with serum galectin-3 concentration (ORs 11.4 and 8.5, respectively). An increased number of vascular complications was associated with high serum galectin-3 levels (P < 0.05). Patients with serum galectin-3 levels > 25 ng/ml had an elevated risk of diabetes relative to patients with levels < 10 ng/ml (OR for any vascular complication 2.64, for heart failure 3.97, for nephropathy 4.09, for peripheral arterial disease (PAD) 4.18; all P < 0.05). Complication risk was higher in patients with neurogenic, stroke, or retinopathy complications, but this difference was not significant after risk factor adjustment. Serum galectin-3 levels correlated with diabetes duration, C-reactive protein (CRP) levels, and albuminuria. CONCLUSION: High galectin-3 values were associated with increased odds of developing heart failure, nephropathy, and peripheral arterial disease in patients with T2DM.

A cross-sectional study on cerebrospinal fluid biomarker levels in cognitively normal elderly subjects with or without a family history of Alzheimer's disease.

BACKGROUND: Cerebrospinal fluid (CSF) biomarkers reflect changes in the brain, and contribute to early screening. Maternal inheritance is putatively stronger than paternal inheritance for late-onset Alzheimer's disease (LOAD). METHODS: Clinical data of 162 cognitively normal subjects were reviewed. A standard questionnaire was used to identify LOAD family history. Mini-mental state examination (MMSE) was used to evaluate cognition. CSF Aß1-40, Aß1-42, total and phosphorylated tau were measured using ELISA. AIMS: To compare biomarkers in cognitively normal elderly subjects with versus without LOAD family history. RESULTS: Among the 162 subjects, 38 and 60 had LOAD family history on paternal and maternal sides, respectively. The remaining 60 subjects had no family history. No difference was noted in age, gender, education level, MMSE score, and memory impairment complaint in the three groups. Aß42 and the Aß42/40 ratio were lower than in subjects with a maternal history than in subjects with a paternal history or without family history (P < 0.05 in both). Phosphorylated and total tau did not differ among the three groups. CONCLUSION: Offspring with a family history of LOAD on the maternal side have lower Aß42 and Aß42/40 ratio in the CSF, and maybe at higher risk for developing AD.

[The relationship between sleep quality and glucose level, diabetic complications in elderly type 2 diabetes mellitus].

OBJECTIVE: To explore the relationship between sleep quality and glucose level, diabetic complications in elderly type 2 diabetes mellitus. METHODS: A total of 130 hospitalized elderly type 2 diabetes in our hospital were included in the study. Questionnaires and other related clinical data were collected within one week after admission. Patients were divided into two groups: poor-sleeper group and good-sleeper group according to Pittsburgh Sleep Quality Index (PSQI). RESULTS: Sixty percent (78/130) of these patients were poor sleepers. The following parameters differed in the two groups: the duration of diabetes [(7.9 ± 1.8) years vs (7.2 ± 1.5) years, t = 2.318], systolic blood pressure [(148 ± 30) mm Hg (1 mm Hg = 0.133 kPa) vs (138 ± 23) mm Hg, t = 2.037], fasting plasma glucose (FPG) [(10.7 ± 2.2) mmol/L vs (9.8 ± 1.9) mmol/L, t = 2.410], hemoglobin A1c (HbA1c) [(8.6 ± 2.2)% vs (7.8 ± 2.1)%, t = 2.068], high-sensitive C-reactive protein (hs-CRP) [(5.27 ± 2.34) mg/L vs (4.44 ± 1.76) mg/L, t = 2.179], ratio of diabetic complications (61% vs 32%, χ(2) = 4.257), percentage of depression (20% vs 8%, χ(2) = 3.722), score of life quality [(98 ± 19) scores vs (89 ± 13) scores, t = 2.980], and proportion of patients treated with insulin (32% vs 12%, χ(2) = 4.489). All the above parameters were significantly higher in poor-sleeper group than the good-sleeper group (all P value < 0.05). Multiple correlation analysis showed that the factors affecting sleep quality were FPG, HbA1c, duration of diabetes, diabetic complications, depression, life quality and insulin application (r = 0.213, 0.257, 0.223, 0.335, 0.422, 0.3451, 0.231, respectively; all P value < 0.05). By multivariate logistic regression analysis, the followings were found: FPG (ß = 1.29, P < 0.05) and PSQI (ß = 1.07, P < 0.05) were found to be correlated with HbA1c. With increasing of PSQI, FPG, HbA1c, diabetic complications and life quality were changed significantly (all P value < 0.05). The independent risk factors of diabetic complications were duration of diabetes (OR = 1.32, 95%CI 1.01 - 2.01), HbA1c (OR = 2.01, 95%CI 1.63 - 2.67), hs-CRP (OR = 1.12, 95%CI 1.08 - 1.21) and PSQI (OR = 1.71, 95%CI 1.58 - 2.02). CONCLUSIONS: Elderly type 2 diabetes mellitus are usually poor sleepers. Sleep quality probably affects blood glucose regulation, and is closely correlated with the occurrence of complications. In addition, poor sleep quality results in poor life quality.

[Caloric restriction suppresses endothelial cells senescence via down-regulation of NOX4 induced by HNF3γ].

The aim of current study is to investigate the molecular mechanism that caloric restriction (CR) suppresses endothelial cells senescence. Human aortic endothelial cells (HAECs) were divided into 5 groups: control group, high caloric group (about 1.5 times caloric intake of control group), low caloric group (about 0.5 times caloric intake of control group), siRNA plus low caloric group (low caloric treatment pretreated with special siRNA targeting hepatocyte nuclear factor 3γ (HNF3γ)), and siRNA plus high caloric group (high caloric treatment pretreated with special siRNA targeting HNF3γ). The gene and protein expressions of HNF3γ and NADPH oxidase 4 (NOX4) were quantified by real-time quantitative PCR (RT-qPCR) and Western blotting, respectively. Intracellular reactive oxygen species (ROS) production was measured by flow cytometry. Endothelial cells senescence was assayed by senescence associated ß-galactosidase staining. After verifying the binding of HNF3γ to NOX4 promoter region by chromatin immunoprecipitation assay (ChIP), NOX4 promoter activity was assayed by dual-luciferase reporter system. Compared with the control group, the mRNA and protein expression levels of HNF3γ,and the ratio of phosphorylated HNF3γ protein increased significantly (P<0.05) in low caloric group, and decreased significantly (P<0.05) in high caloric group and siRNA plus low or high caloric group; whereas the mRNA and protein levels of NOX4 intracellular ROS and endothelial cells senescence decreased significantly (P<0.05) in low caloric group and increased significantly (P<0.05) in high caloric group and siRNA plus low or high caloric group. ChIP result showed there were four HNF3γ binding sites in NOX4 gene promoter region (-6, -76, -249 and -954 bp) and HNF3γ could bind to all 4 predicted sites. According to the results of dual-luciferase reporter system, HNF3γ binding to 1 site (-6 bp), 2 sites (-6 and -76 bp), 3 sites (-6, -76 and -249 bp) and 4 sites(-6, -76, -249 and -954 bp) could suppress NOX4 promoter activity to 80.15±4.64%, 40.02.±2.15%, 16.46±2.24% and 12.13±1.46% compared with that of baseline, respectively ( P<0.05). In a word, low caloric intake decreases the production of intracellular ROS and suppresses endothelial cells senescence through promoting HNF3γ binging to NOX4 promoter region and inhibiting NOX4 gene expression induced by up-regulated HNF3γ.

[Value of tonsillectomy in patient with obstructive sleep apnea hypopnea syndrome with Friedman staging I].

OBJECTIVE: To evaluate the availability of tonsillectomy in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) staged as Friedman I. METHODS: Fifty-six patients with OSAHS in Friedman stage I who refused uvulopalatopharyngoplasty (UPPP) received tonsillectomy merely from January 2004 to March 2010. There were 20 mild, 24 moderate and 12 serious patients respectively in this group. The other 68 OSAHS patients in Friedman stage I received UPPP at the same time as matched group, including 26 mild, 28 moderate and 14 serious patients. RESULTS: There was no significant difference before operation in terms of age, body mass index, apnea hypopnea index (AHI), the lowest pulse oxygen saturation (SPO(2)) and average SPO(2) between the two groups. There were significant difference in mean length of operation (U = 0.000, P < 0.01), hospitalization day (U = 458.5, P < 0.01), visual analogue scale after surgery (U = 0.000, P < 0.01) in these two group. There was no significant difference in surgical effective rate between the two groups (χ(2) = 0.857, P > 0.05). There was also no significant difference in terms of age, body mass index, AHI, the lowest SPO(2) and average SPO(2) after operation between the two groups (t test P > 0.05). The surgical effective rate for the long term of the two groups was equal (χ(2) = 0.857, P > 0.05). Even patients with serious OSAHS in Friedman stage I, the surgical effective rate of the two groups was equivalent (Fisher's exact test, P > 0.05). CONCLUSIONS: Tonsillectomy is a safe and effective surgery for OSAHS in Friedman stage I, whose main structural load lies in the hypertrophic tonsil. It should be the first surgical choice for OSAHS in Friedman stage I.

Predictive value of serum cholinesterase for the prognosis of aged patients with systemic inflammatory response syndrome.

BACKGROUND: Some studies found that cholinesterase (ChE) can be an independent risk factor for patients with multiple organ dysfunction syndrome. To assess aged patients with systemic inflammatory response syndrome (SIRS) early and predict their prognosis, the predictive value of ChE for the prognosis of aged patients with SIRS was analyzed. METHODS: From September 2009 to September 2010, all aged patients with SIRS in the ICU of the Second Affiliated Hospital of Zhejiang University School of Medicine were retrospectively analyzed if they met inclusion criteria: patients aged ≥ 65 years and met American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference criteria for SIRS. Serum ChE, albumin, D-dimer, lactic acid and C-reactive protein (CRP) were measured, and the Acute Physiology and Chronic Health Evaluation (APACHE) II and Glasgow Coma Scale (GCS) scores were evaluated within the first 24 hours in the ICU. Fisher's exact test was used for comparison of the primary disease between the deceased group and surviving group. For comparison of study variables between the two groups, the Student's t test or Mann-Whitney U test was used. Multivariate significance was tested with binary Logistic regression analysis. RESULTS: The clinical data of 124 aged patients with SIRS were collected and analyzed. Sixty-six patients (46 male, 20 female, mean age (78.70 ± 8.08) years) who died were included in the deceased group and 58 patients (34 male, 24 female, mean age (76.02 ± 6.57) years) who survived were included in the surviving group. There were no significant differences in age, gender, APACHE II score and GCS score between the deceased group and surviving group (all P > 0.05), but there were significant differences in lactic acid (P = 0.011), D-dimer (P = 0.011), albumin (P = 0.007), CRP (P = 0.008), and ChE (P < 0.0001). The correlation analysis showed that the APACHE II score and CRP were not correlated with ChE (both P < 0.05). D-dimer and albumin were correlated with ChE (Spearman's rho correlation coefficients were -0.206 and 0.324, the corresponding P values were 0.022 and < 0.0001). Multiple Logistic regression analysis showed that age, gender, lactic acid, D-dimer, albumin, CRP, APACHE II score, and GCS score were not independent risk factors for prognosis of aged patients with SIRS, but that ChE was (P < 0.0001). The receiver operating characteristic curve of ChE had an area under the curve of 0.797 (standard error = 0.04; P < 0.0001), and a ChE of 103.00 U/L was the cut-off value with sensitivity = 0.793, specificity = 0.742. CONCLUSION: Serum ChE might be a predictive marker for the prognosis of aged patients with SIRS, with low serum ChE levels indicating poor prognosis.

[Relationship between plasma total homocysteine and mild cognitive impairment in senile patients with type 2 diabetes].

OBJECTIVE: To explore the relationship between fasting plasma level of total homocysteine (tHcy) and mild cognitive impairment in senile patients with type 2 diabetes mellitus. METHODS: A total of 88 senile type 2 diabetics with mild cognitive impairment treated at our hospital from July 2008 to November 2010 were recruited into the MCI group while 52 senile type 2 diabetics into the DNC group. And the control group was composed of 36 healthy elders. The parameters of tHcy, total glyceride (TG), total cholesterol (TC), low density lipoprotein-C (LDL-C), high density lipoprotein-C (HDL-C), creatinine (Cr), hemoglobin A1c (HbA1c), fasting plasma glucose (FPG), 2 h plasma glucose (2 hPG), fasting insulin (FINS), homeostasis model of insulin resistance (HOMA-IR), folic acid (FA) and vitamin B(12) (VitB(12)) were detected. RESULTS: The patients had a higher level of tHcy in the MCI group than those in the NCM and control groups [(11.3 ± 1.8) vs (9.8 ± 1.5) and (8.1 ± 1.1) µmol/L; P < 0.01]. ROC curve showed that tHcy level had some value of predicting the occurrence of mild cognitive impairment in senile patients with type 2 diabetes mellitus (AUC 0.825, 95%CI 0.758-0.893, P < 0.01). Logistic regression analysis showed that tHcy, SBP, HbA1c, 2 h PG, FINS, LDL-C, HOMA-IR, FA and VitB(12) [OR value: 3.64, 1.68, 1.10, 1.05, 0.81, 1.42, 0.83, 0.74, 0.86 (P < 0.05 or 0.01)] were independent risk factors of mild cognitive impairment in senile diabetic patients. CONCLUSION: Such factors as tHcy, SBP, HbA1c, FPG, 2 hPG, FINS, LDL-C, HOMA-IR, FA and VitB(12) induce senile patients with type 2 diabetes mellitus to suffer mild cognitive impairment. But tHcy level may play an important role in senile diabetic patients with mild cognitive impairment.

[Enhancement of target gene expression by recombinant adeno-associated virus combined with recombinant adenovirus in vivo].

OBJECTIVE: To study the changes in the intensity and temporal pattern of target gene expression in the tumor tissue of nude mice bearing human nasopharyngeal carcinoma (NPC) following injection of recombinant adeno-associated virus (rAAV) and recombinant adenovirus (AdV) in vivo. METHODS: EBV-positive human NPC cell line C666-1 was inoculated subcutaneouly in nude mice. After the tumor mass reached 3 mm in diameter, 1.5 × 10(11) v.g (virus genome) rAAV-EGFP, 2.5 × 10(8) pfu rAdV-EGFP or their balanced mixture was injected intratumorally. At 5 and 10 days after the injection, the tumor tissues were harvested for immunohistochemical staining of GFP, and the ratio of the GFP-positive cells and the intensity of GFP expression was determined. RESULTS: Immunohistochemistry for GFP showed that 5 days after the injection, GFP expression was detected (1.70 ∓ 0.48) in the tumor tissue in rAAV group, and the peak expression levels was seen in rAdV group (6.00∓1.94); the expression level was comparable between the combination group (6.90 ∓ 1.92) and rAdV group. At 10 days, GFP expression was considerably lowered to 2.00 ∓ 0.67 in rAdV group but increased to 8.00∓1.15 in rAAV group. The expression in the combination group maintained a high level at 10 days (10.10∓1.63), which was significantly higher than that in rAAV group (P%0.001). CONCLUSION: Transfection with rAAV combined with rAdV allows instant, sustained and significantly enhanced expression of the target gene in the tumor tissue. This approach takes advantages of the two viruses and can be ideal for exogenous gene delivery into the tumor tissues.

Neferine inhibits angiotensin II-stimulated proliferation in vascular smooth muscle cells through heme oxygenase-1.

AIM: To explore the effect of neferine on angiotensin II (Ang II)-induced vascular smooth muscle cell (VSMC) proliferation. METHODS: Human umbilical vein smooth muscle cells (HUVSMCs) were used. Cell proliferation was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis. Heme oxygenase (HO)-1 protein expression was tested by Western blot analysis. Extracellular signal-regulated protein kinase 1/2 (ERK1/2) activation was determined by using immunoblotting. RESULTS: Pre-incubation of HUVSMCs with neferine (0.1, 0.5, 1.0, and 5.0 micromol/L) significantly inhibited Ang II-induced cell proliferation in a concentration-dependent manner and neferine 5.0 micromol/L increased HO-1 expression by 259% compared with control. The antiproliferative effect of neferine was significantly attenuated by coapplication of zinc protoporphyrin IX (ZnPP IX, an HO-1 inhibitor) with neferine. Ang II-enhanced ERK1/2 phosphorylation was markedly reversed by neferine. By inhibiting HO-1 activity with ZnPP IX, the inhibitive effect of neferine on ERK1/2 phosphorylation was significantly attenuated. Cobalt-protoporphyrin (CoPP), an HO-1 inducer, significantly decreased Ang II-induced ERK1/2 phosphorylation and inhibited Ang II-induced cell proliferation. The ERK1/2 pathway inhibitor PD98059 significantly blocked Ang II-enhanced ERK1/2 phosphorylation and inhibited cell proliferation. CONCLUSION: These findings suggest that neferine can inhibit Ang II-induced HUVSMC proliferation by upregulating HO-1, leading to the at least partial downregulation of ERK1/2 phosphorylation.

[Analysis of clinical characteristics of chronic rhinosinusitis in patients with postirradiated nasopharyngeal carcinoma].

OBJECTIVE: To study the characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma. METHODS: There were 65 cases of chronic rhinosinusitis after irradiated nasopharyngeal carcinoma (NPC, experimental group) and 65 cases of common chronic rhinosinusitis (CRS, control group) in the study. The visual analogue scale (VAS) was used to evaluate the intensity of subjective symptoms. Endoscopic finding was recorded and CT results were evaluated by Lund-Mackay scoring system. RESULTS: As to the VAS, nasal secretion was significantly more severe in experimental group (7.86+/-1.62), compared with control group (5.12+/-1.32, t=10.541, P<0.01). As to endoscopic finding, middle nasal meatus were clean in 35 (53.8%) cases in experimental group, and 23 cases (35.4%) in control group (chi2=4.483, P<0.05). CT score was (7.03+/-4.63) in experiment group, and (11.42+/-3.32) in control group (t=-6.207, P<0.05). The main reason lays in lower CT score and lower involved rate of ostiomeatal complex, frontal sinus, maxillary sinus, anterior ethmoid sinus. CONCLUSIONS: The characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma is quite different from the common CRS and different therapeutic measures should be taken.