To investigate the relationship between the methylation of death-associated protein kinase (DAPK) promoter and gene inactivation in papillary thyroid carcinoma, the technique of methylation-specific polymerase chain reaction (PCR (MSP)) was applied to detect the methylation status of DAPK gene promoter in 70 cases of papillary thyroid carcinoma (study group) and in 50 cases of corresponding adjacent tissues (control group). Immunohistochemical method was used to detect the protein expression; besides, the relationship of DAPK methylation and gene inactivation with pathological factors of papillary thyroid cancer was analyzed. The methylation rate of DAPK was 16% (8/50) in the control group and 71.4% (50/70) in the study group with the difference being statistically significant (χ2 = 19.724, P < 0.01). The methylation of DAPK gene promoter was not associated with age, sex, tumor size, TNM stage, and thyroid capsular infiltration in the study group with papillary thyroid carcinoma (P > 0.05), but was associated with lymph node metastasis (P < 0.05). Spearman’s rank correlation analysis showed that the methylation of DAPK promoter was negatively correlated with the expression of DAPK (r = −0.793, P < 0.01). The methylation of CpG island in the promoter region of DAPK gene can lead to gene inactivation and may be involved in the occurrence of papillary thyroid carcinoma.

Background: The objective of this study was to explore the role of miR-199a-5p in the development of thyroid cancer, including its anti-proliferation effect and downstream signaling pathway.Material/Methods: We conducted qRT-PCR analysis to detect the expressions of several microRNAs in 42 follicular thyroid carcinoma patients and 42 controls. We identified CTGF as target of miR-491, and viability and cell cycle status were determined in FTC-133 cells transfected with CTGF siRNA, miR-199a mimics, or inhibitors.Results: We identified an underexpression of miR-199a-5p in follicular thyroid carcinoma tissue samples compared with controls. Then we confirmed CTGF as a target of miR-199a-5p thyroid cells by using informatics analysis and luciferase reporter assay. Additionally, we found that mRNA and protein expression levels of CTGF were both clearly higher in malignant tissues than in benign tissues. miR-199a-5p mimics and CTGF siRNA similarly downregulated the expression of CTGF, and reduced the viability of FTC-133 cells by arresting the cell cycle in G0 phase. Transfection of miR-199a-5p inhibitors increased the expression of CTGF and promoted the viability of the cells by increasing the fraction of cells in G2/M and S phases.Conclusions: Our study proves that the CTGF gene is a target of miR-199a-5p, demonstrating the negatively related association between CTGF and miR-199a. These findings suggest that miR-199a-5p might be a novel therapeutic target in the treatment of follicular thyroid carcinoma.

It was suggested that muscarinic, and nicotinic receptors increase free Ca[Formula: see text] levels in the facial nerve nucleus via various channels following facial nerve injury. However, intracellular Ca[Formula: see text] overload can trigger either necrotic or apoptotic cell death. It is assumed that, following facial nerve injury, the interactions of nicotinic and muscarinic acetylcholine receptors in facial nerve nucleus may negatively regulate free Ca[Formula: see text] concentrations in the facial nerve nucleus, which provide important information for the repair and regeneration of the facial nerve. The present study investigated the regulatory effects of nicotine on muscarinic receptor-mediated free calcium ion level changes in the facial nucleus in a rat model of facial nerve injury at 7, 30, and 90 days following facial nerve injury using laser confocal microscopy. The dose-dependent regulation of nicotine on muscarinic receptor-mediated free calcium ion level changes in the facial nucleus may decrease the range of free Ca[Formula: see text] increases following facial nerve injury, which is important for nerve cell regeneration. It is concluded that the negative effects of nicotine on muscarinic receptors are related to the [Formula: see text] subtype of nicotinic receptors.

It was suggested that muscarinic, and nicotinic receptors increase free Ca2+ levels in the facial nerve nucleus via various channels following facial nerve injury. However, intracellular Ca2+ overload can trigger either necrotic or apoptotic cell death. It is assumed that, following facial nerve injury, the interactions of nicotinic and muscarinic acetylcholine receptors in facial nerve nucleus may negatively regulate free Ca2+ concentrations in the facial nerve nucleus, which provide important information for the repair and regeneration of the facial nerve. The present study investigated the regulatory effects of nicotine on muscarinic receptor-mediated free calcium ion level changes in the facial nucleus in a rat model of facial nerve injury at 7, 30, and 90 days following facial nerve injury using laser confocal microscopy. The dose-dependent regulation of nicotine on muscarinic receptor-mediated free calcium ion level changes in the facial nucleus may decrease the range of free Ca2+ increases following facial nerve injury, which is important for nerve cell regeneration. It is concluded that the negative effects of nicotine on muscarinic receptors are related to the alpha(4)beta(2) subtype of nicotinic receptors.

The SASH1 (SAM- and SH3-domain containing 1) gene, a member of the SLY-family of signal adapter proteins, has an important regulatory role in tumorigenesis, but its implication in thyroid carcinoma has not been yet investigated. In this study, we investigated the role of SASH1 in proliferation and invasion of thyroid cancer cells and the underlying mechanism. Our results demonstrated that SASH1 is down-regulated in thyroid cancer cells. Overexpression of SASH1 inhibits thyroid cancer cell proliferation, migration and invasion with decreased epithelial-mesenchymal transition (EMT). Mechanistically, overexpression of SASH1 inhibits thyroid cancer cell proliferation and invasion through down-regulation of PI3K and Akt phosphorylation. Taken together, the present study showed that the loss or inhibition of SASH1 expression may play an important role in thyroid cancer development, invasion, and metastasis and that SASH1 may be a potential therapeutic target for the treatment of thyroid cancer.

Background: Several studies demonstrated that the genetic factor contributes a lot to the etiology of hepatitis B virus (HBV) infection. Interleukin 12 (IL-12) plays an important role in the immune response to many viruses. As a functional SNP locus, IL-12B 1188A/C polymorphism (rs3212227) showed an effect on IL-12 production, which may indicate that this polymorphism may influence immune response to HBV during a natural infection.Objective: To investigate whether the IL-12B 1188A/C polymorphism is associated with a risk of HBV infection in the Chinese Han population.Material and methods: Our study was performed on 594 HBV-infected persons and 535 controls. Genotyping procedures for identifying the rs3212227 polymorphism were effected by the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Amplified ribosomal DNA restriction analysis was used to validate genotype results. chi(2) test was used to analyze association analysis.Results: There was no statistically significant association between the IL-12B 1188 A/C polymorphism in HBV-infected persons and controls (for genotypes, chi(2) = 1.976, p = 0.372, df = 2, and for alleles, p = 0.623, OR = 0.959, 95% CI: 0.811-1.134).Conclusions: Our results indicated that the IL-12B1188 A/C polymorphism was not associated with the development of HBV infection in the Chinese Han population.

目的 分析中国近20年侵袭性纤维瘤的流行病学特征及诊疗经验.方法 通过维普数据库和中国知网等多家中文数据库,检索中国近20年有关侵袭性纤维瘤的文章,总结分析侵袭性纤维瘤的流行病学特征和诊治经验.结果 中国近20年共报道侵袭性纤维瘤969例,男女比例约为1∶1.71,平均年龄29岁,高发于10 ～40岁；42.31％分布于华北地区,33.23％分布于华东地区；首发症状中81.23％为无痛或痛性肿块,13.86％为肿瘤压迫引起功能障碍或疼痛；术前诊断为侵袭性纤维瘤仅86例(8.88％),27例于术前行穿刺活检,确诊为侵袭性纤维瘤15例(55.56％)；655例施行根治性手术,95例行姑息性手术,52例术后行放疗,15例于明确诊断后放弃治疗；766例报告随访结果,其中于2个月～8a内复发188例,复发率24.54％.结论 侵 袭性纤维瘤主要分布在华北、华东地区,发病率较低；以无痛或者痛性肿块为主要的临床表现形式,无特异性,术前诊断较困难；治疗以肿瘤一期根治性切除为主,放疗是有效的辅助方法.

Objective: To study the epidemiological characteristics and clinic experience of Biliary papillomatosis.Method: Reports on Biliary papillomatosis published were retrieved from various databases,such as China National Knowledge Infrastructure(CNKI).The epidemiological characteristics and treatment experience of Biliary papillomatosis were analyzed.Results: 39 patients with Biliary papillomatosis were reported in China in recent 30 years,male to female ratio was 1.06:1,with an average age of 60 years.74.36% of the patients located in eastern China,17.24% distributed in the southwest.Abdominal pain and jaundice were the main symptoms.Radical resection of tumor is the most important treatment of Biliary papillomatosis.Conclusion: Biliary papillomatosis mainly locate in the east and southwest.Low incidence and easily misdiagnosed are the characteristics of Biliary papillomatosis.Surgical resection is an effective way to treat the disease.Radical resection of tumor is the most important treatment.

目的 检测阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者血清性激素、一氧化氮(NO)及一氧化氮合酶(NOS)水平的变化,探讨OSAHS患者发生性功能障碍的可能病理生理机制.方法 测定46例男性OSAHS患者和16例健康男性体检者的血清睾酮(T)、性激素结合球蛋白(SHBG)、促黄体生成素(LH)、NO及NOS的水平变化.结果 OSAHS组血清T、LH、NO及NOS水平明显低于对照组,血清SHBG水平高于对照组.差异均有统计学意义(P<0.05).血清T、LH、NO与呼吸暂停低通气指数(AHI)呈负相关,与夜间最低血氧饱和度呈正相关(P<0.05),NOS与AHI呈负相关(P<0.05).血清SHBG与各指标之间无明显相关性(P>0.05).结论 OSAHS患者血清T、LH、NO及NOS水平的变化与病情的严重程度密切相关,在OSAHS导致性功能障碍的发病机制中发挥重要作用.

AIM:To analyze the clinical significance of expression of human telomerase reverse transcriptase (hTERT) and mucin 4 (MUC4) mRNAs in peripheral blood of patients with pancreatic cancer. METHODS:Real-time fluorescence quantitative RT-PCR was applied to investigate the quantitative expression of hTERT and MUC4 mRNAs in peripheral blood of patients with pancreatic cancer and healthy controls. RESULTS:The expression levels of hTERT and MUC4 mRNAs in peripheral blood of patients with pancreatic cancer were significantly higher than those in normal controls (7.95 ± 5.46 vs 0.92 ± 1.07, P < 0.01; 38.25 ± 25.07 vs 4.37 ± 5.96, P < 0.05). The expression of both hTERT and MUC4 mRNAs was associated with lymph node metastasis, surrounding organ infiltration, distant metastasis, and TNM stage (hP = 0.036, 0.027, 0.019; M P = 0.041, 0.022, 0.017). CONCLUSION:Abnormal expression of hTERT and MUC4 mRNAs closely correlates with invasion and metastasis of pancreatic cancer, and hTERT and MUC4 mRNAs in peripheral blood cells might be useful markers for pancreatic caner.

目的：探讨中国胆管乳头状瘤的流行病学特征及临床特点。方法：联合检索维普数据库和中国知网等多家中文数据库中国1979～2011年有关胆管乳头状瘤的文献，总结分析胆管乳头状瘤的临床流行病学特征。结果：共报道胆管乳头状瘤39例，男女比例为1．06：1，平均年龄60岁，高发于50～70岁，74．4％分布于华东地区，17.2％分布于西南地区，以腹痛、黄疸为主要症状。肿瘤根治性切除是最主要的治疗手段。结论：胆管乳头状瘤主要分布在华东、西南地区，发病率较低，术前诊断较困难，易被误诊，手术切除是治疗此病有效方法，手术以肿瘤根治性切除为主。

Objective: To investigate the epidemiological features,diagnosis and treatment of splenic inflammatory myofibroblastic tumor in China during the past 25 years.Methods: The clinical data of 64 patients diagnosed splenic inflammatory myofibroblastic tumor(Splenic inflammatory pseudotumor) published from 1985 to 2010 from various databases,including China National Knowledge Infrastructure(CNKI),VIP,and the epidemiological features,diagnosis and treatment of SIMT patients were retrospectively analyzed.Results: There were 64 patients,including 39 males and 25 females(1.56:1).And their age spread from 24 to 74 with an average of 49.2.Most of patients had not special symptoms,and were diagnosed through routine examinations such as type-B ultrasonic or CT scan.Epigastric pain,debilitation,low-grade fever and some other untypical symptoms might appear in some patients.A solitary splenic lesion might be seen during medical imageologcal examination.Histological types: hypocellular fibrous pattern(n=26),myxoid/vascular pattern(n=12),compact spindle cell pattern(n=5).Immunophenotypic findings show that the spindle cells were positive for Vimentin,SMA in most cases.For most patients,splenectomy were the mainly treatment,and there were no signs of relapse and metastasis after the operation.Conclusion: Splenic inflammatory myofibroblastic tumor is extremely rare in china,and there are not special clinical symptoms or signs.It can be identified by characteristics of medical imaging,histologic and immunophenotypic findings with other splenic lesions.But it is difficult to define its nature preoperatively.Splenectomy is the mainly treatment with a satisfactory prognosis.

OBJECTIVE To study the location of facial nerve and prevent facial nerve injury in middle ear surgery according to dissection of temporal bone and experience of middle ear surgery. METHOD Thirty sides of temporal bones were exposed tympanic and mastoid segment of facial nerve with facial nerve decompression. The course of facial nerve was located by the markers of middle ear. RESULT Tympanic segment of facial nerve passed between horizontal semicircular canal and stapes,then superior and anterior to the cochleariform process. Mastoid segment of facial nerve located in posterior wall of tympanic cavity. The mastoid segment of facial nerve travelled below the level of horizontal semicircular canal and annulus membrane tympani, and the extension line of its posterior margin and posterior-one-third of horizontal semicircular canal intersected to form an included angle (117.04 +/- 2.42) degrees. External genu of facial nerve located anterior and inferior to the horizontal semicircular canal. The shortest distance was (1.97 +/- 0.53) mm between middle point of horizontal semicircular canal and facial nerve, (1.03 +/- 0.29) mm between incus short process and facial nerve, (0.93 +/- 0.25) mm between cochleariform process and facial nerve, (1.18 +/- 0.42) mm between head of stapes and facial nerve, (3.08 +/- 0.28) mm between tympani sulcus and facial nerve at the vestibule window level, and (2.13 +/- 0.34) mm between tympani sulcus and facial nerve at round window level, respectively. CONCLUSION Horizontal semicircular canal carina, incus short process, stapes, annulus membrane tympani and cochleariform process are ideal landmarks of tympanic and mastoid segment of facial nerve, which are helpful in middle ear surgery.

Muscarinic receptors and nicotine receptors can increase free calcium ion levels in the facial nucleus via different channels following facial nerve injury. In addition, gamma-aminobutyric acid A (GABA(A)) receptors have been shown to negatively regulate free calcium ion levels in the facial nucleus by inhibiting nicotine receptors. The present study investigated the influence of GABA(A), gamma-aminobutyric acid B (GABA(B)) and C (GABA(C)) receptors on muscarinic receptors in rats with facial nerve injury by confocal laser microscopy. GABA(A) and GABA(B) receptors exhibited significant dose-dependent inhibitory effects on increased muscarinic receptor-mediated free calcium ion levels following facial nerve injury. Results showed that GABA(A) and GABA(B) receptors negatively regulate muscarinic receptor effects and interplay with cholinergic receptors to regulate free calcium ion levels for facial neural regeneration.

Previous studies have demonstrated that the cholinergic system via nicotinic receptors regulates intracellular free calcium levels in the facial nucleus under normal physiological conditions However the regulation of nicotinic receptors on free calcium levels following facial nerve injury remains unclear In the present study an animal model of facial nerve injury was established and changes in nicotinic receptor expression following facial nerve injury in rats were detected using reverse transcription polymerase chain reaction Nicotinic receptor mediated changes of free calcium levels following facial nucleus injury were determined by laser confocal microscopy Results showed no significant difference in nicotinic receptor expression between the normal group and the affected facial nerve nucleus The nicotinic receptor alpha(4)beta(2) subtype increased free calcium levels following facial nerve injury by promoting calcium transmembrane influx, and L type voltage gated calcium channel mediated influx of calcium ions played an important role in promoting calcium transmembrane influx The nicotinic receptor mediated increase of free calcium levels following facial nerve injury provides an important mechanism for the repair of facial nerve injury

The muscarinic receptor modulates intracellular free calcium ion levels in the facial nerve nucleus via different channels In the present study, muscarinic receptor-mediated free calcium ions levels were detected by confocal laser microscopy in the facial nerve nucleus following facial nerve injury in rats There was no significant difference in muscarinic receptor expression at the affected facial nerve nucleus compared with expression prior to injury, but muscarinic receptor-mediated free calcium ion levels increased in the affected side following facial nerve Injury (P < 0.01) At day 30 after facial nerve injury, 50 mu mol/L muscarinic-mediated free calcium ion levels were significantly inhibited at the affected facial nerve nucleus in calcium-free artificial cerebrospinal fluid, and the change range was 82% of artificial cerebrospinal fluid (P < 0 05) These results suggest that increased free calcium ion concentrations are achieved by intracellular calcium ion release, and that the transmembrane flow of calcium ions is also involved in this process

Previous studies have demonstrated that muscarinic, and nicotinic receptors increase free Ca2+ levels in the facial nerve nucleus via various channels following facial nerve injury. However, intracellular Ca2+ overload can trigger either necrotic or apoptotic cell death. Gamma-aminobutyric acid (GABA), an important inhibitory neurotransmitter in the central nervous system, exists in the facial nerve nucleus. It is assumed that GABA negatively regulates free Ca2+ levels in the facial nerve nucleus. The present study investigated GABA type A (GABA(A)) receptor expression in the facial nerve nucleus in a rat model of facial nerve injury using immunohistochemistry and laser confocal microscopy, as well as the regulatory effects of GABA(A) receptor on nicotinic receptor response following facial nerve injury. Subunits alpha 1, alpha 3, alpha 5, beta 1, beta 2, delta, and y3 of GABA(A) receptors were expressed in the facial nerve nucleus following facial nerve injury. In addition, GABA(A) receptor expression significantly inhibited the increase in nicotinic receptor-mediated free Ca2+ levels in the facial nerve nucleus following facial nerve injury in a concentration-dependent fashion. These results suggest that GABA(A) receptors exhibit negative effects on nicotinic receptor responses following facial nerve injury.

Background Ca2+ in the central nervous system plays important roles in brain physiology, including neuronal survival and regeneration in rats with injured facial motoneurons. The present research was to study the modulations of intracellular free Ca2+ concentrations by cholinergic receptors in rat facial nucleus, and the mechanisms of the modulations.Methods The fluorescence intensity of facial nucleus in Fluo-3 AM loaded acute brainstem slices was detected by applying intracellular free Ca2+ measurement technique via confocal laser scanning microscope. The changes of fluorescence intensity of facial nucleus indicate the average changes of intracellular free Ca2+ levels of the neurons.Results Acetylcholine was effective at increasing the fluorescence intensity of facial nucleus. Muscarine chloride induced a marked increase of fluorescence intensity in a concentration dependent fashion. The enhancement of fluorescence intensity by muscarine chloride was significantly reduced by thapsigargin (depletor of intracellular Ca2+, store; P<0.01), rather than Ca2+ free artifical cerebrospinal fluid or EGTA (free Ca2+ chelator; P>0.05). And the increase of fluorescence intensity was also significantly inhibited by pirenzepine (M-1 subtype selective antagonist; P<0.01) and 4-DAMP (M-3 subtype selective antagonist; P<0.01). In addition, fluorescence intensity was markedly increased by nicotine. The enhancement of fluorescence intensity by nicotine was significantly reduced by EGTA, nifeclipine (L-type voltage-gated Ca2+ channel blocker), dihydro-beta-erythroidine (alpha 4 beta 2 subtype selective antagonist), and in Ca2+ free artificial cerebrospinal fluid (P<0.01), but not in the presence of mibefradil (M-type voltage-gated Ca2+ channel blocker) or thapsigargin (P>0.05).Conclusions The data provide the evidence that muscarinic receptors may induce the increase of intracellular free Ca2+ levels through the Ca2+ release of intracellular Ca2+ stores, in a manner related to M, and M3 subtypes of muscarinic receptors in rat facial nucleus. Nicotine may increase intracellular free Ca2+ concentrations via the influx of extracellular Ca2+ mainly across L-type voltage-gated Ca2+ Channels, in a manner related to the alpha 4 beta 2 subtype of nicotinic receptors. Chin Med J 2009; 122(9):1049-1055

OBJECTIVE:To establish the visual patch clamp whole-cell recording technique and study the properties and functional significance of muscarinic receptor-mediated currents in rat medial vestibular nucleus neurons (MVNn). METHODS:Brain slices containing the MVN were prepared from fifteen Wistar rats. By combining infrared differential interference contrast (IR-DIC) technique and CCD-Camera system with visual patch clamp whole-cell recording technique, twenty healthy neurons were located and muscarinic receptor-mediated currents in rat MVNn were observed and analyzed. RESULTS:Visual patch clamp technique can be used to make direct localization and to make sure of active neuron. In MVNn, a comparison of the current-voltage relationships before and during the application of muscarine, which revealed an increase in the slope of the I-V curve and the reversal potential for this response lay at (-88.4 +/- 4.9) mV (x +/- s), indicates that the activation of muscarinic cholinergic receptors leads to a decrease in potassium current. The test in the voltage sensitivity of the muscarine-induced response, which showed that the effect had a linear current-voltage relationship and reversed at (-86.7 +/- 3.5) mV, indicates that the potassium current blocked by muscarine corresponds to the voltage-insensitive leak potassium current. CONCLUSIONS:Visual patch clamp technique, which was considered better than blind patch clamp technique, can improve the success of sealing process. By the analysis of muscarinic receptor-mediated currents, the data provide support that muscarinic cholinergic mechanisms play a prominent role in the modulation of the excitability of MVNn and also offer a new idea for the efficacy of anticholinergic drugs.

咽旁间隙解剖部位深在,结构复杂,上至颅底,下达舌骨平面.咽旁间隙内有颈内动脉、颈外动脉、颈内静脉、交感神经、第Ⅸ～Ⅻ对脑神经、颈深上淋巴结群等重要结构.发生于咽旁间隙的肿瘤较为少见,约占头颈部肿瘤的0.5%[1,2],由于其位置深在,周围结构复杂,术前定性诊断较难.细针穿吸细胞学检查(fine needle aspiration cytology,FNAC)以其准确、快速、安全、经济的优势,广泛应用于乳房肿块、腮腺肿块、甲状腺肿块等疾病的诊断,为影像学检查不可替代的一项有价值的诊断方法.FNAC在咽旁间隙肿瘤诊断中的应用价值,目前尚存争议[1,3,4].笔者分析我科收治的术前行FNAC的63例咽旁间隙肿物患者的临床、病理资料,以探讨FNAC对咽旁间隙肿瘤的诊断价值.