Background p16INK4a gene plays an important role during the aging and senility.So it was well known to be a leading gene associated with aging.Corneal endothelial cells(CECs) always get trapped in the G1 phase due to the lack of proliferative ability.Whether it is relative with cell senescence is unclear.Objective This study was to investigate the expression ofp16INK4a gene and Bmi1 gene in human CECs from different aged donors ex vivo.Methods The corneal rims,the residual of corneal tissue preserved in DX solution after penetrating keratoplasty,was used in the present study.Parameters were recorded for the donor,including the age,death to preservation interval and preservation to surgery interval.Corneal endothelium survival rate and endothelial cell density were evaluated by trypan blue-alizarin red dying immediately after penetrating keratoplasty.Routine haematoxylin and eosin staining was also performed to proof the normal structure of the cornea.Sections of corneas from different aged donors were classified into ＜30 years group,30-50 years group and ＞50 years group and were immunostained to assess the expressions of p16INK4aprotein,Bmi1 protein and Ki67 protein in CECs.Total RNA was extracted from independent corneal sample for the evaluation of p16INK4a mRNA,Bmi1 mRNA and Ki67 mRNA expression in CECs by quantitative real-time PCR(qRT-PCR).Results The endothelial cell density of each group was (3069 ±172),(2748±64),(2444 ±178)cells/mm2,respectively.Haematoxylin and eosin staining showed the normal structure of corneal epithelium,stroma and endothelium.qRT-PCR examination revealed an age-related increase in p16INK4a mRNA expression in the CECs(F =5.703,P =0.014) and a decrease in Bmi1 mRNA and Ki67 mRNA expression (F =3.950,P =0.042;F=548.500,P =0.000).The further comparison verified a significant elevation in the expression of p16INK4a mRNA in the CECs of the ＞50 years group compared with ＜30 years group and significant decline in Bmil mRNA and Ki67 mRNA the expression (P =0.006,0.013,0.000).Immunohistochemistry in situ confirmed the expression and nuclear localization of p16INK4a protein in CECs,and the expressing intensities of Bmi1 and Ki67 proteins in the elder donors were weaker than those of the younger donors.The immunofluorescence exhibited that the expressing intensity of p16INK4a protein in CECs of 58 years old donor was higher than that of 23 years old donor,showing a consistent result with that of qRT-PCR.Conclusions Expression of p16INK4a gene increases and that of Bmi1 gene decreases upon age.These results suggest that p16INK4a gene is associated with senescence of human CECs.

Blast Injuries ; Deafness ; etiology ; Humans

?Color vision has been an important part of the human visual function.It is determined by the phytochrome of cone.In many clinical cases of ocular fundus diseases, patients had color vision loss, which shows that color visions tests is necessary and meaningful.At present, doctors have not paid attention to it and adopt the tests rarely. The article summarizes the tests of color in common use and applications on the diagnosis and treatment for common ocular fundus diseases to be a theory basis.

ObjectiveThe possibility of the hyperplasi a of prostatic outer gland was studied with transrectal ultrasound (TRUS) guided biopsies. MethodsTwenty-seven prostatic outer glands in patients suspicious for prostatic cancer (PCA) were biopsied by TRUS guided, the total sites were 47. The same site was biopsied in sagittal and longitudinal section respectively. The hypoechoic lesion was biopsied 2 times if it was discovered in the outer gland. The histological results were diagnosed in double blind. Results Twenty prostatic inner glands looked symmetrical hyperplasia; 9 outer glands became thin because of the compression, there was no compressive sign in 18 patients; there was a hypoechoic lesion in outer gland of 3 patients. Benign prostatic hyperplasia was diagnosed in the specimens of 47 sites. ConclusionsThe hyperplasia of prostatic outer gland can occur like inner gland because it is glandular tissue.

Objective To evaluate the clinical value of repeat transrectal ultrasound (TRUS) guided biopsy in prostatic cancer (PCA). Methods Repeat TRUS guided biopsy was conducted on 54 patients of the previous benign TRUS guided biopsy at high risk of PCA. Postatic specific antigen ranged in 0.5- 90.0 ?g/L(mean 16.3 ?g/L). Twenty-nine patients were abnormal in digital rectal examination, 21 patients were abnormal in TRUS. Results Forty-seven patients had 2 consecutive times of the biopsy, 6 patients had 3 consecutive times, 1 patient had 4 consecutive times in 54 patients of repeat TRUS guided biopsy. The pathological finding was confirmed in 14 PCA ( 25.9%), and 31 benign prostatic hyperplasia, 5 prostatic intra-epithelial neoplasmia, 4 chronic prostatitis. Conclusions The detection of PCA will be increased by repeat TRUS guided biopsy at high risk of PCA after the previous benign TRUS guided biopsy.

Objective To study the characteristic of the hyperplasia hypoechoic nodules in outer gland of prostate with transrectal ultrasound (TRUS) guided biopsies. Methods 42 patients were suspected of prostatic cancer (PCa) for the existence of hypoechoic nodules in their outer glands of prostate. Each nodule was TRUS guided biopsied in sagittal and longitudinal section, respectively. The histological examinations were performed in double blind. Results Among the 42 patients, there were 22 cases of benign prostate hyperplasia (52.4%) and 20 cases of prostate cancer (47.6%). Conclusions The sonographic findings of the hypoechoic nodules in outer glands included not only PCa, but also benign hyperplasia nodules. Therefore, the definitive diagnosis must dependent on pathologic examinations.

Objective To clarify the clinical significance of p21 WAF1 and the relationship between it and p53. Methods Samples of human cholangiocarcinoma (HC) tissue and paired normal bile duct tissue adjacent to the tumor from 30 patients with HC were employed in this study. In situ hybridization (ISH) was used to detect the p53 and p21 mRNA expression. Immunohistochemistry (IHC) was applied to analyze p53 gene mutation and P21 protein expression. Results p21 WAF1 And p53 gene protein expression was detected in 36.7% (11/30) and 53.3% (16/30) of the carcinoma specimens, respectively, and none of the paired tissue by IHC. p53 Positivity was related to local lymph node metastasis. Comparing with non-lymph node metastatic group, p53 positivity in metastatic group was significantly higher. p53 Positivity of cholangiocarcinoma in clinical stage III was significantly higher than that in clinical stages I and II. The survival time was significantly shorter in patients with P53 protein expression than in those without. It was found that p53 expression was not associated with p21 WAF1 expression. Conclusions P53 Positivity may be correlated with tumor development but not tumor occurrence. The fact that p53 expression was not associated with p21 WAF1 expression indicates that p53-independent activation of p21 WAF1 may exist.

AIM: To establish the methods for determining ephedrina hydrochloridum,pseudoephedrine hydrochloride,puerarin,baicalin in Ganmao Soft Capsule (Herba Ephedrae,Radix Puerariae lobatae,Radix Scutellariae,Ramulus Cinnamomi,Radix Angelicae sinensis,Radix Angelicae dahuricae,etc.) METHODS: Part 1(determination of ephedrina hydrochloridum and pseudoephedrine hydrochloride):The HPLC was carried out on kromasilTMC18 column(4.6 mm ? 250 mm,5 ?m) with acetonitrile-0.02 mol/L phosphoric acid solution(pH2.7)-triethylamine(2 ∶ 98 ∶ 0.1)as mobile phase,gradient elution.The detection wavelength was set at 207 nm.Part 2(determination of pueranin and baicalin):The HPLC was carried out on kromasilTMC18 column(4.6 mm ? 150 mm,5 ?m)with acetonitrile-water(11 ∶ 89)as mobile phase,gradient elution.The detection wavelength was set at 250 nm.RESULTS: The average recoveries were 99.96% with RSD 1.25% ,98.41% with RSD 1.46% ,98.67% with RSD 0.91% ,99.30% with RSD 0.88% ,respectively.CONCLUSION: The assay results demonstrate that the methods are simple,accurate and reproducible for quantification of four active components in Ganmao Soft Capsule.

Objective To observe the effect of case-based learning (CBL) applied in ward-round on residents training. Methods A total of 33 residents trained with routine ward-round and CBL ward-round in the neurological department were investigated with questionnaire. Results 30 effective answers were recalled. More of them reported that CBL ward-round was very significant in improving clinical thinking, mastering the specialized knowledge, developing autonomous learning and improving the communication with patients and their family members (P<0.05). Conclusion CBL ward-round is effective on improving the comprehensive professional abilities of residents.

Objective To investigate the proliferation inhibition and apoptosis induction effect of thalidomide(Thal) and its joint effect with 5-Fu on MGC-803 cell line. Methods The morphological changes of MGC-803 cells with AO/EB stain were observed under fluorescence microscope. The proliferation inhibition effeet was evaluated with MTT. The apoptosis induction effect was determined by FAM. Results The results of MTr array were as below: the difference was significant between Thai groups of 25 mg/L or above concentration and the control group (P＜0.05), 5-Fu of all testing concentrations showed significant difference compared with the control gToup(P＜0.01). For combined groups with the same concentration of 25 mg/L 5-Fu,combined groups showed distinct difference from the corresponding 5-Fu group (P＜0.01), but two combined groups showed no distinct difference with each other (P＞0.05). For combined groups with the same concentration of 5-Fu 12.5 mg/L, 5-Fu plus 50 mg/L or 100 mg/L Thai showed no distinct difference from 5-Fu group of 25 mg/L(P＞0.05). With FAM study, all test groups showed significant difference compared with the control group (P＜0.01). There was also significant difference between two test groups arbitrarily (P＜0.01).Conclusion Both Thai and 5-Fu could inhibit the proliferation and induce the apoptosis of MGC-803 cells.The effect is reinforced with the combination significantly. Combination with Thai could decrease the concentration of 5-Fu but have the similar effect.