| Objective:The study aimed to explore the feasibility of urine Raman spectroscopy in the diagnosis of postmenopausal women with osteoporosis.Methods:Postmenopausal women were selected as the subjects,and were divided into osteoporosis group(n=30),osteopenia group(n=27)and normal bone mass group(n=28)according to the results of dual energy X-ray examination.We collected urine and serum samples from all subjects.Urine surface enhanced Raman spectroscopy(SERS),serum sex hormone levels,serum lipid levels and biochemical markers of bone turnover markers were detected.Single factor analysis of variance,principal component analysis and linear discriminant analysis were used to compare the Raman spectra.Results:1.Characteristics of subjectsThe age,BMI,serum sex hormone,bone metabolism index and blood lipid index of all the subjects were in normal distribution,and single factor analysis of variance showed that there was no significant difference in BMI,testosterone,TP1NP,osteocalcin,total vitamin D and LDL.The age of osteoporosis group and normal bone mass group were(67.72 ± 11.50)and(57.46 ± 10.23)years old,the difference was statistically significant(p<0.01).E2 in osteoporosis,osteopenia and normal group was(26.43± 13.51)pg/mL,(36.68±17.22)pg/mL and(45.89±27.50)pg/mL respectively,CTX,one of the bone turnover markers in three group was(0.55 ±0.25)ng/mL,(0.48 ± 0.22)ng/mL and(0.36 ± 0.22)ng/mL respectively,and the difference was statistically significant(p<0.01).Lipid index TG and TC of osteoporosis group were(1.92±1.51)mmol/L and(4.72±1.13)mmol/L,osteopenia group were(1.83±1.11)mmol/L and(4.56±0.99)mmol/L and normal group were(1.22±1.92)mmol/L and(3.74±1.34)mmol/L,the difference was statistically significant(p<0.01).There was a significant difference in HDL between the bone loss group,(1.20±0.27)mmol/L,and the normal group,(1.35±0.32)mmol/L,(p<0.01).The above results showed that age,estrogen,bone turnover and serum lipid levels had a certain effect in the development of osteoporosis.2.Comparison of three groups of urine Raman spectroscopyBy comparison of smoothing and normalization of the normal bone mass group,the average Raman spectrum of osteopenia and osteoporosis group had no significant deviation,as a strong peak intensity at 1000cm-1 displacement as an example,the three groups had no statistical significance.A total of 10 characteristic peaks were found in the range of 200 cm-1-2000 cm-1,located at 254 cm-1,450 cm-1,576 cm-1,723 cm-1,786 cm-1,894 cm-1,1000 cm-1,1462 cm-1,1598 cm-1,1715 cm-1.The average peak intensity of the osteoporosis group was weaker than that of the normal bone mass group before 1000 cm-1,and the Raman line was shifted downwards.There was no significant difference between the spectroscopy of the bone loss group and the normal bone mass group after 1000 cm-1,while the osteoporosis group increased the intensity,the line shift.The substance each peak represented was mainly lipid and protein,which was consistent with the increase of TC,TG and LDL in blood.At 723 cm-1,the peak intensity of osteoporosis or bone loss group was significantly different from that of normal bone mass group(p<0.01).At 786 cm-1,there was a difference in the intensity of Raman spectroscopy between osteoporosis and osteopenia(p<0.05)and at 1000 cm-1,there was a difference between the osteopenia group and the osteoporosis or normal group(p<0.05).3.Comparison of relative intensity of I723 between osteoporosis group and normal bone mass groupThe relative peak intensity(crest/trough)of 58 subjects in the osteoporosis group and the normal bone mass group at the characteristic peak at 723 cm-1 was calculated and the scatter plot was plotted based on the results.There were differences in the distribution of scatterplots between the two groups,with a total of 1.14 as the cut-off value.Most of the patients with osteoporosis were under it,while the urine Raman spectra I723/I705 were greater than 1.14 in most normal bone mass group.The discriminant analysis was used to confirm the results.The results were correctly diagnosed in 25 cases of osteoporosis patients,5 cases were misdiagnosed;19 cases of 28 cases of normal group made the correct diagnosis,9 cases were misdiagnosed as osteoporosis.The sensitivity and specificity of the diagnosis were 83.3%and 67.9%,respectively.The accuracy rate was 75.9%and the accuracy rate was low.It can be seen that the relative peak intensity of the Raman spectrum at 723 cm-1 can be used to distinguish and differentiate the osteoporosis patients and healthy people.However,because of its less spectral information,it is not enough to distinguish the two sets of data,which can be used as the basis for discrimination.4.PCA-LD A Results of urine Raman spectroscopy3 groups of subjects,a total of 85 cases with respective Raman spectra of 723cm-1 Raman peak as reference peak normalized for the other 9 characteristic peaks and the PCA score of normalized eigenvalue of each case was calculated.The 2 principal components with eigenvalues greater than 1 were obtained,and the cumulative contribution rate was 91.94%.To calculate the first,the second principal component score making scatter diagram and the results showed that it was difficult to distinguish between the osteoporosis group and the bone mass reduction group,while the normal bone mass group had obvious difference line with the other two groups.According to the PCA score for discriminant analysis and examination,the sensitivity of diagnosis of osteoporosis was 66.7%,the specificity was 92.9%,the overall accuracy was 79.3%;the sensitivity and specificity of diagnosis of bone loss were 63.0%and 92.9%respectively,and the accuracy of the overall discrimination was 78.2%.Conclusion:1.There was a significant difference in urine Raman spectroscopy between the osteopenia or osteoporosis group and the normal bone mass group,which was mainly reflected in the change of intensity without significant displacement.2.The biomolecule information provided by the urine Raman spectrum of the bone loss change groups is consistent with the physiological and biochemical changes of the disease.3.The detection of urine Raman spectrum can be used to prompt osteoporosis and osteopenia,but it is not easy to distinguish between osteoporosis and osteopenia,its application in clinical pending further study. |
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