OBJECTIVE: To investigate the expression of major histocompatibility complex class I chain-related gene A (MICA) in laryngeal squamous cell carcinoma(LSCC), and its clinical significance. METHOD: Immunohistochemistry and RT-PCR were used to detect the expression of the MICA in LSCC and normal tissue samples. The relationship between the expression of MICA and the clinicopathologic features features were was analyzed. RESULT: Compared to the expression of MICA in normal tissues samples, the expression of MICA in LSCC tissue was significantly increased (P < 0.01). MICA expression level in carcinoma tissue was closely related to the tumor-differentiation degree and TNM staging. CONCLUSION Our study suggests that MICA may play an important role in the invasion and metastasis of LSCC, and could be a potential tumor maker for LSCC.
Aged ; Carcinoma ; genetics ; pathology ; Female ; Histocompatibility Antigens Class I ; genetics ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Real-Time Polymerase Chain Reaction

Objective To investigate the therapeutic effects of titanium artificial auditory ossicles in closed type mastoid radical tympanoplasty ,in comparison with ceramic artificial auditory ossicles .Methods A retrospec‐tive analysis was done on of 150 patients (155 ears) with the closed type of mastoid radical tympanoplasty using arti‐ficial auditory ossicles for hearing reconstruction (from January 2008 to December 2012) ,cund were ,and followed up for 12 months .Based on the two kinds of artificial auditory ossicle materials ,the group was divided into two :122 cases (127 ears) for the titanium auditory ossicle group and 28 cases (28 ears) for the ceramic auditory ossicle group .We compared the average air conduction hearing threshold and air bone gaps (ABG) at 0 .5 ,1 .0 ,2 .0 ,and 4 .0 kHz ,at preoperative 6 months and postoperative 6 ,12 months .Results For the Titanium artificial auditory os‐sicle group:117 ears (92 .13% ) had hearing threshold improvement >15 dB HL ,106 ears of the ABG change ≤20 dB ,which were statistically significant .The total success rate of hearing reconstruction was 83 .46% (106/127) . For the ceramic auditory ossicle group:5 ears (17 .86% ) had hearing threshold improvement >15 dB HL ,3 ears (10 .71% ) of the ABG change ≤20 dB ,which was no statistical significance between the two groups .Conclusion The hearing reconstruction effects of titanium artificial auditory ossicle in closed type mastoid radical tympanoplasty is excellent .It is suitable for application in closed-mastoidectomy tympanoplasty ideal artificial auditory ossicle .

ObjectiveTo investigate the feasibility of examining aortic pulse wave velocity (PWV),aortic distensibility (AD) and brachial artery flow-mediated dilation (FMD) by means of highresolution 3.0 T MRI.MethodsA total of 32 healthy volunteers underwent high-resolution MRI to assess aortic PWV,and AD in ascending aorta (AA),proximal descending aorta (DA),distal descending aorta (DDA) and FMD of the brachial artery with repeat examination performed in 1-2 hours.PWV was evaluated by 2D Phase Contrast (PC) velocity-encoded MRI with a 4.7-7.8 ms temporal resolution.Fiesta-cine MRI was used to assess AD and FMD with a 18.75-31.25 ms temporal resolution.The image quality of these two scans was scored and the agreement between them was tested with Kappa analysis.The reproducibility of the results between repeated measurements of PWV,AA-AD,DA-AD,DDA-AD and FMD was assessed with intra-class correlation coefficient (ICC) analysis.The method of Bland-Altman plot was used to assess the agreement between results of repeated studies.Results Each examination including PWV,AD and FMD were completed in about half an hour.The image quality between repeated scans showed good agreement ( Kappa value 0.776 ) with the score of ( 3.53 ± 0.62 ) and ( 3.41 ± 0.67 ) respectively.Reproducibility between repeated measurements was high for aortic PWV [ (4.33 ± 0.88 ) vs ( 4.36 ±0.88) m/s],AA-AD [(8.60±3.11) × 10-3 vs (8.59 ± 3.10) × l0-3/mm Hg(1 mm Hg =0.133 kPa) ],DA-AD[ (6.95 ±2.44) × 10-3 vs (6.95 ±2.42) × 10-3/mm Hg],DDA [(10.54 ±2.91) ×l0-3 vs (10.55 ±2.90) × 10-3/mm Hg] and FMD [(24.94 ± 12.55)％ vs (24.92 ±1 2.38 ) ％ ].ICC were 0.95,0.97,0.99,0.98 and 0.94,P ＜ 0.01.Excellent agreement between repeated measurements was found for aortic PWV [ confidence interval (CI) between - 0.55 and 0.50 ],AA-AD ( CI between - 0.11 and 0.12 ),DA-AD ( CI between - 0.08 and 0.08 ),DDA-AD ( CI between - 0.23 and 0.21 ) and FMD (CI between - 1.46 and 1.51 ).The maximum difference percentage in minimum average for aortic PWV,AA-AD,DA-AD,DDA-AD and FMD was 38.53％,9.65％,3.86％,5.68％,42.37％,respectively,all less than 50％.Conclusion Comprehensive assessment of aortic compliance and brachial endothelial function can be achieved using 3.0 T high-resolution MRI with excellent reproducibility and within a reasonable amount of time.

Objective To investigate the imaging features of focal nodular hyperplasia and hepatocellular carcinoma on DWI. Methods The data of patients with histopathologically confirmed FNHs and HCCs between August 2008 and November 2010 were collected. A total of 24 patients with 26 FNH lesions and 36 patients with 39 HCC lesions were included in our study. All patients underwent breath-hold DWI with b = 500 s/mm2 and dynamic contrasted-enhanced (DCE) MRI. The imaging findings of FNHs and HCCs were retrospectively analyzed and compared. The signal intensity (SI) of the lesions on DWI were classified as iso-, slightly high, high SI and the distribution of SI between FNHs and HCCs was compared with Fisher exact test. ADC value and lesion-to-liver ADC ratio of FNHs and HCCs were measured and compared by using independent sample t test. ROC was performed to assess the diagnostic value of ADC value and lesion-liver ADC ratio in the characterization FNHs versus HCCs. Results Of 26 FNHs,23 manifested as isointensity or slightly high SI on DWI, but most 25 out of 39 HCCs showed high SI. The distribution of SI between FNHs and HCCs had significant difference ( P = 0. 000). The mean ADC value and lesion-liver ADC ratio for FNHs [ (1.76 ± 0. 62 ) × 10-3 mm2/s and 1.06 ± 0. 18, respectively ] were significantly higher ( P = 0. 001, P = 0. 000, respectively ) than those for HCCs [ ( 1.26 ± 0. 46 ) × 10-3mm2/s and 0. 79 ±0. 12, respectively]. The area (Az) under the ROC for the ADC value and lesionliver ADC ratio for the differentiation of FNHs versus HCCs were 0. 79 ± 0. 05 and 0. 85 ± 0. 05,respectively, with no significant difference (P =0. 270). The specificity of the two measures was 69. 23% and 97.44%, respectively, with significant difference (P = 0. 001 ). Conclusion FNH shows isointensity or slightly high SI with relatively higher ADC value and lesion-liver ADC ratio than those of HCCs on DWI,which is characteristic for its diagnosis and differentiation.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

AIM: To investigate the effects of protein C activator (PCA) from Agkistrondon acutus venom ( AAV) on the tension of thoracic aorta rings isolated from the rats with sepsis.METHODS:The model of sepsis was es-tablished by intraperitoneal injection of lipopolysaccharide ( LPS) .SD rats were randomly divided to 6 groups ( n=6 ):sham group, LPS group, PCA intervention group (LPS+PCA, PCA at doses of 0.1 mg/kg, 0.3 mg/kg and 0.6 mg/kg) and LPS+polymyxin B (at dose of 0.2 mg/kg) group.Using perfusion experiment in vitro, the tension of the aortic rings was measured by biological signal analytical system.RESULTS:The values of MABP, HR, LVDP and ±dp/dtmax were significantly lower in LPS group than those in sham group and LPS+PCA groups.Compared with sham group, the relaxa-tion response to acetylcholine ( ACh) and the contractile response of aorta rings induced by phenylephrine ( Phe) were sig-nificantly decreased in LPS group, which were increased significantly in PCA intervention group ( especially at dose of 0.6 mg/kg) compared with LPS group.The dose-response curve of aorta contraction with denuded endothelium induced by Phe shifted down significantly in LPS group compared with sham group, and no significant difference between LPS group and PCA intervention group was observed.Also no statistical difference was found in non-endothelium dependent relaxation of aortic rings induced by sodium nitroprusside among the groups.Pretreatment of N-nitro-L-arginine methl ester and methyl-ene blue increased the contraction amplitude of aortic rings induced by Phe.CONCLUSION:PCA from AAV effectively reverses the hypoergia of the vessels in rats with sepsis through protecting vascular endothelium, the mechanism of which may be mediated by inhibiting NO-GC-cGMP signal transduction pathway.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

AIM: To study the basic mechanism of transcriptional regulation,NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested.METHODS:(1.04 kb)-promoter fragment of NKX3.1 gene was obtained by PCR and cloned into pGL_3-basic and pEGFP-1 that are promoter-less reporter vectors to examine its promoter activity driving the reporter gene transcription.The promoter activity was determined by dual-luciferase reporter assay and the expression of GFP reporter observed under fluorescence microscope.RESULTS: The sequence of the cloned(1.04 kb) promoter proved to be correct by DNA sequencing.The dual-luciferase reporter assay(M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL_3-(1.04 kb) promoter was about 1.5-fold higher than that of pGL_3-control transfection and 50-fold higher than that of pGL_3-basic transfection.To investigate the 1.04 kb-promoter activity in different tumor cell lines,the constructed pGL_3-(1.04 kb) promoter and pEGFP-1.04 kb promoter were transfected into several cell lines,respectively.The results showed that the activity of(1.04 kb) promoter in LNCaP was highest among the tested cell lines.Multiple consensus sequence elements have been identified within the(1.04 kb) fragment using TRANSFAC database.Further experiments will be done to determine their founctions.CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

OBJECTIVE: To investigate the leukotriene D4 synthase gene A (LTD4S A)-444 C polymorphism in persistent allergic rhinitis (AR) of Chinese Han nationality and to evaluate its relevance to clinical responsiveness of leukotriene receptor antagonist. METHOD: There were 150 patients [87 males, 63 females, average age (38 +/- 14)] diagnosed with persistent AR in Allergy clinic in our hospital from March 2010 to March 2012; 146 healthy controls (78 males, 68 females, mean age (39 +/- 12)). We detected LT D4SA-444C polymorphism and allele frequencies with Polymerase Chain Reaction (PCR) and-Restriction Fragment Length polymorphism (RELP). The treatment group received monotherapy leukotriene receptor antagonist (montelukast) for 4 weeks. Urinary leukotriene D4 (LTD4) levels were detected by enzyme-linked immunosorbent assay (ELISA) before and after treatment, respectively. We evaluated anti-leukotriene treatment response according to the changement of symptoms, signs PTS and urinary LTD4. We tested correlation between LT D4S gene-444C allele frequency and the treatment response by multivariate analysis of variance. RESULT: (1) LTD4S gene-444 genotype AA/CC, AC/CC frequency is 70.7% (106/150) and 29.3% (44/150), allele A, C frequencies is 67.3% (101/150) and 32.7% (49/150) in AR group, and LTD4S gene-444 genotype AA/CC, AC/CC frequency is 76.7% (112/146) and 23.3% (34/ 146), allele A, C frequencies is 74.0% (108/146) and 26.0% (38/146) in healthy control group, there is not statistically significant difference between two groups. (2) Among 150 AR patients, compared to patients with AA/CC genotype, the genotype AC/CC patients are younger [average age (35 +/- 9), and (50 +/- 18) respectively, F = 5.891, P < 0.05], with earlier age of onset [(31 +/- 4), and (46 +/- 6) respectively, F = 6.985, P < 0.05], longer course of disease [(8.7 +/- 2.1), and (3.1 +/- 2.0) respectively, F = 11.43, P < 0.05], higher symptom scores (8.2 +/- 0.2; 4.8 +/- 0.3), higher signs score (7.3 +/- 3.3; 3.4 +/- 5.1), and the difference was statistically significant. (3) After 4 weeks of montelukast treatment in AR patients, treatment response of anti-leukotriene in genotype AC/ CC patients is better than those in AA/CC genotype patients (F = 11.01, P < 0.05), the differences of treatment response between two groups were correlated with LTD4 levels in vivo, clinical symptoms and signs of patients. CONCLUSION In a Chinese Han population the LTD4SA-444B polymorphism might be one of the factors in the clinical response to leukotriene receptor antagonists in persistent AR patients.
Adult ; Arachidonate 5-Lipoxygenase ; genetics ; Case-Control Studies ; Female ; Gene Frequency ; Humans ; Leukotriene Antagonists ; therapeutic use ; Male ; Middle Aged ; Polymorphism, Genetic ; Rhinitis, Allergic ; drug therapy ; genetics ; Young Adult