| Background and objective:Gastroesophageal reflux disease(GERD)is a common disease,counting for nearly 11~25 % of digestive specialist outpatients.GERD with typical symptoms presents heartburn and reflux,counting for almost 40~60% of entire patient community.While,the proportion of symptoms manifesting as cough,hoarseness,asthma and pharyngitis,is about 70%~90% in extra-esophageal symptoms,which often mislead to respiratory or throat problems.Nowadays,GERD attracts profound attentions because of the soaring number of gastroesophageal patients due to diet diversification,growing metal stresses and less-regulated food administrations.Current diagnostic methods of GERD were summarized as below,including 24-hour pH monitoring,high-resolution esophageal manometry(HRM),endoscopy,gastrointestinal barium meal imaging,symptom-related questionnaire and PPI diagnostic treatment.Nonetheless,each method shows rather limitation,for instance,most approaches are invasive,costly and poor tolerance for treated patients.Previous studies demonstrated that the main content and injury factors in reflux fluid contained pepsin,majority in type-III,acid and bile.The pepsin III complex accounts for 80%,which is closely related with GERD.Current available diagnostic protocols emphasized on discerning symptoms,acid/bile in reflux and motility of LES,rather than advanced molecule based detection.Here,we developed an applicable analytical platform based on immunoreaction to quantitatively determine the pepsin in saliva/sputum,obviating the need of invasive sampling and drastically reducing the cost for analysis.Research on pepsin detection for the diagnosis of GERD or throat reflux is a hot spot in Europe nowadays,but rare reported in China.Besides,people in Chongqing are fond of spicy,alcohol and tobacco,which can increase the incidence of GERD.So it is of great significance to carry out this study in Chongqing.Subject:250 GERD patients(male 84,female 166,age: range,24 to 70,mean age: 45.38 ± 0.74 years)from Daping Hospital were divided into following groups: Typical symptoms GERD group(44 cases,male 12,female 32,age: range,24 to 63,mean age: 43.10 ± 1.70 years),including NERD group(41 cases,male 10,female 31,age: range,27 to 63,mean age: 43.29 ± 1.73 years;NERD with 24 h pH monitoring group(50 cases,male16,female 34,age: range,25 to 61,mean age: 43.10 ± 1.54 years);Typical symptoms GERD with proton pump inhibitor(PPI)treatment(46 cases,male 18,female 28,age: range,24 to 70,mean age: 44.85 ± 1.63 years);Extra-esophageal symptoms GERD group(24 cases,male 9,female 15,age: range,25 to 62,mean age: 43.83 ± 2.19 years);Extra-esophageal symptoms GERD with PPI treatment(19 cases,male 4,female 15,age: range,24 to 63,mean age: 45.42 ± 3.04 years);Typical combined with extra-esophageal symptoms GERD group(19cases,male 9,female 10,age: range,25 to 63,mean age: 46.79 ± 2.68 years);Barrett’s esophagus group(24 cases,male 13,female 11,age: range,28 to 68,mean age: 50.50 ± 2.19 years);GERD with anxiety and depression group(27 cases,male 4,female 23,age: range,28 to 61,mean age: 44.74 ± 1.77 years),and healthy controls(26 cases,male 10,female 16,age: range,25 to 68,mean age: 48.38 ± 2.17 years).Method:Montreal consensus on gastroesophageal reflux disease and GERD questionnaire was administered to recruit the typical symptoms GERD group;GERD questionnaire and endoscopy was used to recruit Non-erosive reflux disease(NERD)group;Endoscopy was used to pick out Barrett’s esophagus group;The Hamilton Anxiety and Depression Scale was conducted to evaluate GERD with anxiety and depression group.To detect the pepsin III concentration in saliva samples by Colloidal gold double-antibody sandwich method;24h pH monitoring technique and SPIII detection was conducted to study the diagnostic value of NERD comparatively,percentage of total time of pH <4.0 and DeMeester score in esophagus was recorded to analysis 24 hours acid reflux.To analysis the differences of SPIII level by SPSS20.0 Nonparametric test,and the differences of gender and positive rates among each group were analyzed by chi-square test.The differences of ages among each group were analyzed by one-way analysis of variance.Results:1.The difference of SPIII level was found between sub-types of GERDSPIII concentration in group of typical symptoms [ 359.5(18.25-500)ng / mL] was significantly higher than that in controls [ 16(16-31.5)ng / mL](Z=3.816,P = 0.000);SPIII concentration in group of extra-esophageal symptoms [ 142(16-356.75)ng / m L] was also higher than that in controls(Z=2.037,P = 0.042);SPIII concentration in group of typical combined with extra-esophageal symptoms [ 83(16-356)ng / mL] increased significantly,Compared with the control group(Z=2.220,P = 0.026).The positive rate in group of typical symptoms was significantly higher than that of controls [70.45%(31/44)vs.15.38%(4/26),χ2 = 19.825,P < 0.001];and the positive rate in group of extra-esophageal symptoms was higher than controls [54.17%(13/24)vs.15.38%(4/26),χ2 = 8.365,P = 0.004];the positive rate in group of typical combined with extra-esophageal symptoms was also higher than that of controls [52.63%(10/19)vs.15.38%(4/26),χ2 = 7.106,P = 0.008].SPIII concentration in group of typical symptoms [ 359.5(18.25-500)ng / mL] increased significantly than that measured in GERD with anxiety/depression [ 16(16-138)ng / mL](Z=4.020,P = 0.000);The positive rate of typical symptoms group was also significantly higher than that of GERD with anxiety/depression [70.45%(31/44)vs.[29.63%(8/19),χ2 = 11.264,P = 0.001].2.PPI diagnostic test on SPIII level in GERD patients with typical symptoms and extra-esophageal symptoms.Compared with the group of typical symptoms GERD [ 359.5(18.25-500)ng / mL],SPIII concentration in typical symptoms GERD with PPI treatment [ 16(16-337)ng / m L] was significantly lower(Z=3.096,P = 0.002),and the positive rate of typical symptoms GERD with PPI treatment was also significantly lower [70.45%(31/44)vs.34.78%(16/46),χ2 = 11.469,P = 0.001].SPIII concentration in group of extra-esophageal symptoms GERD with PPI treatment [ 16(16-46)ng / mL] was significantly lower than extra-esophageal symptoms GERD group [ 142(16-356.75)ng / mL](Z=1.993,P = 0.046),and the positive rate in group of extra-esophageal symptoms GERD with PPI treatment was also significantly lower than that of extra-esophageal symptoms GERD group [ 54.17%(13/24)vs.21.05%(4/19),χ2 = 4.864,P = 0.027].Compared with controls,there was no significant difference in SPIII levels of GERD with extra-esophageal and typical symptoms after PPI treatment(P > 0.05).3.Comparative study of 24 h pH monitoring and SPIII testing for the diagnosis of NERDResult of 24 h pH monitoring: of the 50 NERD patients,22 had pathological acid reflux(percentage of pH < 4.0 exceeds 4.2% and/or DeMeester score exceeds 14.72).The overall positive rate was 44%(22/50).Result of SPIII detecting: Of the forty-four NERD patients,29 was positive.The overall positive rate was 70.73%.SPIII detection has a higher positive rate than that of 24 h pH monitoring for NERD patients [70.73%(29/41)vs.44%(22/50)(χ2 = 6.535,P = 0.011)].Conclusions:1.Our findings showed a higher prevalence and concentration of salivary pepsin in patients with typical and/or extra-esophageal symptoms group compared with GERD patients with anxiety and depression and healthy controls,suggesting salivary pepsin testing as an effective intrinsic maker for diagnosis of sub-types of GERD.2.Pepsin III levels and positive rates were not significantly altered i n GERD patients with anxiety and depression,indicating that mental and psychological factors play an important role in the occurrence of reflux symptoms in such patients.This may lessen the use of unnecessary antireflux therapy.3.PPI diagnostic testing can significantly reduce the SPIII levels and positive rates of GERD patients with extra-esophageal symptoms and typical symptoms,suggesting that PPI may reduce the production of pepsin by inhibiting gastric acid secretion to increase the pH value in stomach.4.For NERD patients,the positive rate of salivary pepsin III was significantly higher than 24 h pH monitoring,suggesting that the two methods can complement with each other to improve the diagnostic efficiency of NERD.Salivary pepsin testing may complement to assist office-based diagnosis.This may lessen the need for further invasive and expensive diagnostic methods. |
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