The Expression Of TGFβ₁, Ki67 In Benign Proliferative Vocal Cord Tissues And Research About Correlation Between Laryngeal Function And Vocal Cord Pathology

ObjectivesNearly 10 years, with the development of phoniatrics, there are more and more attentions of clinicians, and phoniatrics gradually had been expanded from a simple means of scientific research to quantitative analysis means of laryngeal vocal function and clinical research of voice diseases. As accelerating the pace of life and work, different physiological stages (Voice change time, menopause), endocrine, mental emotional factors, environmental factors such as dust in the air, we have been finding more and more patients with hoarseness, and finding that vocal cord polyps and vocal cord nodules are the most common cause of hoarseness. Laryngocarcinoma is one of the most common malignant tumorof head and neck carcinoma in recent years, its incidence is rising, and it is a serious threat to human life. The development from benign lesions of the vocal cords to throat cancer is a gradual evolution process, so the potential forecast about malignant transformation of laryngeal precancerous lesions is very important. The basic pathological features of laryngeal precancerous lesions is epithelial dysplasia, and therefore the study of laryngeal epithelium from normal to malignant cell proliferation during the transition process, may reflect the degree of laryngeal epithelial hyperplastic lesions. In recent years, the research about relation between transforming growth factor-β1(TGFβ1)and tumor occurrence is fairly active. Transforming growth factor and tumor occurrence, development and tumor cell proliferation, differentiation and apoptosis are closely related. TGFβ1 as a pleiotropic growth factor, is expressed as inhibition of laryngeal cancer cell cycle-associated antigen Ki67 (Ki-67 Antigen) is a non-protein nuclear antigen in proliferation nucleus, and it is currently the best detected indicator of cell proliferation activity. Study about TGFβ1, Ki67 in benign hyperplasia vocal fold lesions (including benign and precancerous lesions) aspects and their correlation is less. This study attempts to find indicators of voice and vocal cord pathology correlation, filtering and effective voice parameters and immunohistochemical indicators for contracting the disease, provide the basis for prognosis. This study attempts to find indicators of voice and vocal cord pathology correlation, filtering effective voice parameters and immunohistochemical indicators so as to provide the basis for determine the condition andr the prognosis.We mant toserve the following purpose:1. From the molecular level to discuss the pathogenesis of voice disease;2. To provide objective diagnostic evidence for quantifying voice disease and determing the extent of voice disorders-an objective, scientific, data based evaluation;3. Attempt to establish an objective voice, multi-parameter assessment model;4. To search the intrinsic link between voice disorders'pathology and laryngeal function tests;5. To provide the basis for determing whether vesting or prevention of the disease-investigating laryngeal precancerous lesions;6. To provide basis for observation and treatment evaluation to prevent recurrence after the operation.MethodsBy inclusion, exclusion criteria we have randomly selected these pathological paraffin from December 2007 to October 2010.According to pathology outcome these cases were divided into three groups:vocal cord polyp group with cases, mild dysplasia group with 38 cases, moderate dysplasia group with 11 cases. Age:18 years-65 years of age (including 18 and 65 years) with an average age of 50.28 years; Gender:male 36, female 40 cases; course:1-123 months,12 months or less 35 cases,29 cases in 12-60 months and 12 cases in 60 months or moreWe have done research on organization pathology and clinical research. On the one hand, we did organization pathologic research, prepared pathological slice by conventional method, the two-step EnVision detection system were performed to carry out TGFβ1, Ki67 immunohistochemical staining, image acquisition conditions were fixed, then we have combined the average percentage of positive cells and the level scores of staining intensity to develop one semi-quantitative scoring method- H integral, We have respectively recorded TGFβ1, Ki67 expression in the three H points. For statistical purposes, I have further divided H points into 4 H-level points: 0 (0, negative),1 (1-100 points, weakly positive),2 (101-200, medium positive),3 (201-200, strongly positive). We have orderly done TGFβ1, Ki67 H expression analysis in each pathological group. And in each pathology group we have done correlation analysis between TGFβ1 and Ki67.The other research is Clinical research, fixed check conditions, we have entried preoperative and postoperative voice acoustic parameters (fundamental frequency perturbation Jitter, amplitude perturbation Shimmer, normalized noise energy NNE, harmonic to noise ratio HNR, Maximum hponation time MPT, S/Z), electronic EGG parameters (Jitter, Shimmer, NNE, HNR, Contact quotient perturbation CQP, Contact quotient CQ), stroboscopic laryngoscopy subjective shape evaluation parameters (mucosal wave, phase closure), doctor of psychological auditory evaluation parameters (total hoarse voice G, rough sound R, breath sound B). The two parameters-mucosal wave,closure phase are divided into three levels:0 is full loss of function,1 is partly loss of function and 2 is functioning properly. The three parameters G, R, B are divided into four levels:0, normal,1 mild,2 moderate,3 severe. In each pathological group we contrasted parameters before and after the surgery, and within each pathological group analysed the correlation of each kind of parameters。Result1. Research on organization pathology(1) Expression of Ki67 in different pathology groups:The positive rate in each group were significantly different compared (p <0.001). Polyp group, the weakly positive rate is highest, morethan 18.5%; mild atypical hyperplasia, the weakly positive rate is also main, reaching 57.9%;moderate atypical hyperplasia, the strongly positive rate is highest in positive rate, reaching 36.4%.In the different pathology grouping, the Ki67 expression positive rate assumes along with the pathological change degree's aggravation advances gradually the tendency. Especially polyp group and the mild non-typical group masculine gender rate difference is obvious. After the card side examination, the difference has statistics significance (p<0.05)(2)Expression of TGFβ1 in different pathology groups:The positive rate in each group were significantly different compared (p<0.001). Polyp group, the medium positive rate is highest, reaching 40.7%; mild atypical hyperplasia, the medium positive rate is main, reaching 63.2%; moderate atypical hyperplasia, the mainly positive rate is weakly positive rate, reaching 45.5%.In the different pathology groups, TGFβ1 the expression masculine gender rate also advances gradually along with the pathological change degree's aggravation, and the difference is obvious. After the card side examination, the difference has statistics significance (p<0.001)(3)Each group separately analyzes the expression relevance between Ki67 and TGFβ1:①The polyp group:Ki67 expression was positively correlated with the expression of TGFβ1, the correlation coefficient was notstatistically significant, r=0.227, p=0.255;②Mild atypical proliferation group:The Ki67 expression assumes the inverse correlation relations with TGFβ1 the expression, but the correlation coefficient has non-statistical significance, r=-0.216, p=0.193;③Moderate atypical proliferation group:The Ki67 expression assumes the remarkable inverse correlation relations with TGFβ1 the expression.The correlation coefficient was statistically significant, r=-0.760, p=0.007。(4)The three groups'distribution characteristic at age, the sex, course It is not every obvious.2. Clinical research(1)In different pathology groups voice acoustics test result comparison has indicated that there is non-statistics significance (P>0.05) in three group with computer acoustics results before the surgery.(2)In different pathology groups voice acoustics test result comparison has indicated that by comparing the three groups after the technique, between mild non-typical groups and the polyp group, polyp group S/Z surpasses the mild non-typical proliferation group explicitly, and there is statistics significance (p<0.05) (3)Voice acoustics test result comparison between before and after technique, computer analysis of speech sound each parameter has the distinct improvement besides S/Z, the difference is remarkable, particularly at Jitter, Shimmer, the NNE aspect the difference is remarkable (p≤0.001).(4) In different pathology groups the electroacoustical gate chart test result has indicated Shimmer, CQP dropped after the surgery contrasting by before the surgery, the difference is very remarkable (respectively is p<0.003,p<0.002).Also about Jitter,HNR, the difference is very remarkable (respectively is p=0.001,p<0.05)(5)In erery pathology grouping aroud surgery the frequency dodges laryngoscope subjective shape appraisal result comparison after technique, the mucous membrane wave has been distinct improved, the glottis closed situation also has been changed for the better, Statistics significance remarkable (P<0.001)。(6) In erery pathology groups psychological sense of hearing appraisal comparison by before and after surgery has showed polyp group G and mild non-typical proliferation group B obtained distinct improvement, it is remarkable statistics significance (P≤0.001);(7) Voice acoustics test EGG test relevance analysis:In the various parameters of computer voice test the relevance among EGG, HNR, Jitter, Shimmer and NNE is better(p<0.01), especially with HNR best(p<0.001).(8) Each group analyze the psychological sense of hearing appraisal and voice acoustics test result relevance separately:①Polyp group,R and Jitter, Shimmer, the HNR relevance is remarkable, has statistics significance (p<0.001)②The mild non-typical proliferation group, Jitter, Shimmer and G, R relate closely(p<0.001).③the moderate non-typical proliferation group, only the Jitter and the sound quality relations are close, particularly always hoarse, has the remarkable statistics significance(p<0.001).(9) Each group analyze the psychological sense of hearing appraisal and the electroacoustical gate chart test result relevance separately:①Polyp group G and Jitter, Shimmer, NNE, CQ have the relevance, positive related, after the rank related examination, has statistics significance (p<0.05) ②Mild non-typical proliferation group relevance of G and Jitter, HNR is obvious, being positive related with the former, with the latter inverse correlation. All has the remarkable statistics significance (p<0.01)③Moderate non-typical group, in all correlation coefficients, G and the Jitter relevance is high, but non-statistics significance.Conclusion1. Research on organization pathology(1)Ki67 becomes sensitive at forecasting pathological malignant change, and has the specificity. Carry out fashionable Ki67 expression inspection when you do pathology biopsy may get clear diagnosis direction, may judge disease's prognosis extension to turn over.(2)The expression relationship of Ki67 and TGFβ1 in the three groups: polyps group showed a positive correlation between the two; mild and moderate dysplasia showed a negative correlation, especially in moderate dysplasia, a negative correlation between the two relations significantly.(3)Pathological studies show that in the transition process from polyp to the mild and moderate dysplasia, Ki67 expression rate's gradually increasing tips in all activities of the cell cycle (G1, S, G2 and mitosis) cell proliferation activities have been gradually increasing. The pathogenesis of laryngeal precancerous lesions might be G1, S, G2 and mitosis period enhanced cell proliferation.(4)Not only TGFβ1 has participated in the fiber structure proliferation in intercellular substance of the vocal cord polyp organization but also possibly participated the same activity in intercellular substance of the lightly and the moderate atypical proliferation organization. It is suggested that in the mild and moderate atypical proliferation pathological change cell fiber structure's proliferation is also one of basic pathology changes. And for mild and moderate dysplastic lesions fibroplasia was significantly more than polyps lesions(5) In this study, the function of TGFβ1 at inhibition of malignant transformation and inhibition of epithelial cell proliferation is not obvious.2. Clinical Research(1)Obey the perioperative clinical pathway on troditional chinese medicine chronic hoarseness we treat the vocal cord benign proliferation diseases remarkable effectively, and in improveming vocal cord vibrational frequency, the scope, the acoustic fidelity, the glottis closed situation various aspect curative effect is remarkable.(2)In the polyp group, Jitter, Shimmer, HNR, CQ, G, R are the sensitive targets. G is more serious, the vocal cord vibrational frequency irregularity is bigger, the scope fluctuation irregularity is bigger.(3)In the mild atypical proliferation group, Jitter, Shimmer, HNR, G, R are more sensitive. G is always more serious, the vocal cord vibrational frequency irregularity is bigger, the harmonics ingredient is less; R is more serious, the vocal cord vibrational frequency irregularity is bigger, the vocal cord closing time is longer, the harmonics ingredient is less.(4)In the moderate atypical proliferation group, Jitter, G are only slightly sensitive.(5)Three pathology groups in the respective parameter relevance analysis we have discovered that sense of hearing psychology appraisal's G is most sensitive, the stable test target. Acoustics test's Jitter, Shimmer, HNR are sensitive, stable. Especially the mild atypical proliferation group, G, Jitter, HNR are especially close in the relations. While in moderate atypical proliferation group, eliminates G and Jitter, other various voices parameter is insensitive.(6) Unify Jitter, Shimmer, HNR, G, R -ulti parameter analysis may provide evidence for judging the condition, the curative effect, the reexamination along with the target.(7)The voice parameters are sensitive, but the specificity is not strong, so they are unable to provide the accurate basis for disease diagnosis. However, they can be used as an objective basis to judge the clinical effect...