Objective To investigate the changes of macrophages and hemophagocytes in bone marrow smears of patients with multiple myeloma(MM)before and after chemotherapy and their correlation with clinical prognostic value.Methods A total of 300 MM patients treated in Liaocheng Second People's Hospital Affiliated to Shandong First Medical University from June 2018 to June 2023 were selected as the study objects.All patients received at least 3 courses of chemotherapy and were divided into the remission group(n=214)and the non-remission group(n=86)according to the clinical effect.Immunoglobulin(Ig)A,IgG,CD3+,CD4+,CD8+,interleukin(IL-2),IL-4,IL-6,IL-17,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β,macrophages and hemophagocytes were detected in the two groups and compared between the two groups.COX regression was used to analyze the relationship between immunological indexes and non-remission of chemotherapy.The relationship between macrophages and hemophagocytes and non-remission of chemotherapy was analyzed by restricted cubic spline.The multiplicative interaction of macrophages and hemophagocytes on non-remission of chemotherapy was analyzed using an unconditioned Logistic regression model,and the additive interaction was analyzed using the interaction calculation table.Receiver operating characteristic(ROC)curve analysis of macrophages and hemophagocytes alone or in combination to determine the value of chemotherapy non-remission.Results The overall response rate(ORR)and non-response rate(NRR)of MM patients were 71.33%and 28.67%respectively.Compared with before treatment,IgA,IgG,CD8+,IL-6,IL-17,TNF-α,TGF-β,macrophages and hemophagocytes were significantly decreased in both groups after treatment(tslow=9.252～61.177,tnot slow=4.057～35.797).CD3+,CD4+,CD4+/CD8+,IL-2 and IL-4 were significantly increased(tslow=9.706～33.940,tnot slow=4.227～16.167),and the differences were statistically significant(all P<0.05).After treatment,compared with the remission group,IgA,IgG,CD8+,IL-6,IL-17,TNF-α,TGF-β,macrophages and hemophagocytes in the non-remission group were significantly higher than those in remission group(t=3.362～30.028),CD3+,CD4+,CD4+/CD8+,IL-2 and IL-4 were significantly lower than those in remission group(t=3.736～13.998).and the differences were statistically significant(all P＜0.05).After adjusting the influence of other factors by COX regression,the trend test of macrophages and hemophagocytes was still statistically significant the trend test of macrophages and hemophagocytes was still statistically significant(P<0.05).Patients with≥5 macrophages/tablet and≥3 hemophagocytes/tablet had a significantly increased risk of non-remission from chemotherapy(P<0.05).There were additive(OR=6.157,95%CI:3.768～12.978)and multiplicative(OR=5.648,95%CI:1.035～17.492)interactions between macrophages and hemophagocytes in the non-remission of chemotherapy.Compared with the single judgment of macrophages and hemophagocytes,the combination of the two has the highest accuracy in determining chemotherapy non-remission(P＜0.05).Conclusion Macrophages and hemophagocytic cells in MM patients after chemotherapy are significantly lower than those before chemotherapy,with≥5 macrophages/tablet and≥3 hemocyte phages/tablet,indicating that the risk of non-remission in patients with chemotherapy increased significantly,and the combination of the two can accurately judge the clinical efficacy.

Objective To investigate the changes of macrophages and hemophagocytes in bone marrow smears of patients with multiple myeloma(MM)before and after chemotherapy and their correlation with clinical prognostic value.Methods A total of 300 MM patients treated in Liaocheng Second People's Hospital Affiliated to Shandong First Medical University from June 2018 to June 2023 were selected as the study objects.All patients received at least 3 courses of chemotherapy and were divided into the remission group(n=214)and the non-remission group(n=86)according to the clinical effect.Immunoglobulin(Ig)A,IgG,CD3+,CD4+,CD8+,interleukin(IL-2),IL-4,IL-6,IL-17,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β,macrophages and hemophagocytes were detected in the two groups and compared between the two groups.COX regression was used to analyze the relationship between immunological indexes and non-remission of chemotherapy.The relationship between macrophages and hemophagocytes and non-remission of chemotherapy was analyzed by restricted cubic spline.The multiplicative interaction of macrophages and hemophagocytes on non-remission of chemotherapy was analyzed using an unconditioned Logistic regression model,and the additive interaction was analyzed using the interaction calculation table.Receiver operating characteristic(ROC)curve analysis of macrophages and hemophagocytes alone or in combination to determine the value of chemotherapy non-remission.Results The overall response rate(ORR)and non-response rate(NRR)of MM patients were 71.33%and 28.67%respectively.Compared with before treatment,IgA,IgG,CD8+,IL-6,IL-17,TNF-α,TGF-β,macrophages and hemophagocytes were significantly decreased in both groups after treatment(tslow=9.252～61.177,tnot slow=4.057～35.797).CD3+,CD4+,CD4+/CD8+,IL-2 and IL-4 were significantly increased(tslow=9.706～33.940,tnot slow=4.227～16.167),and the differences were statistically significant(all P<0.05).After treatment,compared with the remission group,IgA,IgG,CD8+,IL-6,IL-17,TNF-α,TGF-β,macrophages and hemophagocytes in the non-remission group were significantly higher than those in remission group(t=3.362～30.028),CD3+,CD4+,CD4+/CD8+,IL-2 and IL-4 were significantly lower than those in remission group(t=3.736～13.998).and the differences were statistically significant(all P＜0.05).After adjusting the influence of other factors by COX regression,the trend test of macrophages and hemophagocytes was still statistically significant the trend test of macrophages and hemophagocytes was still statistically significant(P<0.05).Patients with≥5 macrophages/tablet and≥3 hemophagocytes/tablet had a significantly increased risk of non-remission from chemotherapy(P<0.05).There were additive(OR=6.157,95%CI:3.768～12.978)and multiplicative(OR=5.648,95%CI:1.035～17.492)interactions between macrophages and hemophagocytes in the non-remission of chemotherapy.Compared with the single judgment of macrophages and hemophagocytes,the combination of the two has the highest accuracy in determining chemotherapy non-remission(P＜0.05).Conclusion Macrophages and hemophagocytic cells in MM patients after chemotherapy are significantly lower than those before chemotherapy,with≥5 macrophages/tablet and≥3 hemocyte phages/tablet,indicating that the risk of non-remission in patients with chemotherapy increased significantly,and the combination of the two can accurately judge the clinical efficacy.

Objective:To investigate the association of single nucleotide polymorphism at the estrogen receptor 1(ESR1) gene rs1801132 with the risk of brick-tea type skeletal fluorosis.Methods:The typical brick-tea type fluorosis areas in Qinghai, Xinjiang, and Inner Mongolia were selected as the survey sites for a cross-sectional study. An epidemiological questionnaire was conducted by the staffs on the sites for participants older than 16 years, and physical examination and X-ray diagnosis were performed. Brick tea, blood, and urine samples were collected at the same time. The diagnosis of skeletal fluorosis through X-ray was based on the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS/T 192-2008); The determination of tea's fluoride and urinary fluoride was performed by fluoride ion-selective electrode method; gene sequencing analysis of rs1801132 locus of ESR1 gene was done by Sequenom MassARRAY flight mass spectrometry system.Results:A total of 994 patients were included in this study. The total prevalence of skeletal fluorosis was 23.9% (238/994). The prevalence of skeletal fluorosis in Tibetans(39.9%, 123/308) was higher than those of Mongolian and Han nationality [22.2% (58/261), 13.4% (57/425), χ 2=20.435, 67.811, P < 0.05]. Based on binary logistic analysis, the daily tea fluoride intake ≤ 3.5 mg, urinary fluoride content ≤1.6 mg/L, and age ≤45 years were used as the reference groups, and then, when the daily tea fluoride intake > 7.0 mg ( OR=2.865, 95% CI: 1.923-4.268), urinary fluoride content > 1.6-3.2 mg/L ( OR=2.368, 95% CI: 1.686-3.326) and > 3.2 mg/L ( OR=3.559, 95% CI: 2.401-5.276), the age > 45-65 years old ( OR=2.361, 95% CI: 1.603-3.477) and > 65 years old ( OR=4.556, 95% CI: 2.845-7.296), the risk of fluorosis was higher than that of the reference group, respectively. When the daily tea fluoride intake was > 3.5-7.0 mg and the level of urinary fluoride was > 1.6-3.2 mg/L, G allele had a protective effect on skeletal fluorosis in Mongolian population (adjusted OR=0.207, 95% CI: 0.044-0.974); when the daily tea fluoride intake was > 3.5-7.0 mg, gender was male group, G allele had a protective effect on skeletal fluorosis in Han population (adjusted OR=0.315, 95% CI: 0.112-0.887). Conclusion:The single nucleotide polymorphism of the rs1801132 locus at the ESR1 gene may be associated with the risk of susceptibility to brick-tea type skeletal fluorosis in Mongolian and Han nationality.

Objective To explore the distribution of serum Mg,A1,Ca,Fe,Zn and Cu ions in Tibetan population in drinking tea type fluorosis areas,and to provide a clue for pathogenesis of drinking tea type fluorosis.Methods Tibetans from six villages in Qinghai Province (Maqin County and Dari County),who were over 16 years old,born and grew up in those villages,were included.All of the participants were received epidemic questionnaire survey,tea water samples were collected and the fluoride concentration was tested based on the standard of Brick Tea Fluoride Content (GB 19965-2005).Meanwhile,the daily amount of brick tea consumption was surveyed to calculate the daily intake of tea fluoride.Blood samples were collected and the concentration of serum Mg,Al,Ca,Fe,Zn and Cu ions was tested by the method of inductively coupled plasma mass spectrometry (ICP-MS).All of the participants were diagnosed by X-ray,the parts we scheduled were forearm,shank and pelvic,then the skeletal fluorosis was diagnosed based on the Diagnostic Criteria for Endemic Fluorosis (WS/T 192-2008).Results A total of 170 people were surveyed,74 people were skeletal fluorosis,and 96 people were non-skeletal fluorosis.The median (quartiles) of daily intake of tea fluoride was 7.35 (3.00,12.30) mg.The concentration of serum Mg ion was 22.02 (17.30,23.67) mg/L,Al ion was 0.22 (0.14,0.38) mg/L,Ca ion was 100.03 (88.56,112.73) mg/L,Fe ion was 1.66 (1.26,2.36) mg/L,Zn ion was 0.80 (0.63,0.95) mg/L and Cu ion was 1.28 (0.99,1.48) mg/L.The concentration of serum Fe ion was higher in male [2.13 (1.37,3.09) mg/L] than that of female [1.56 (1.18,2.02) mg/L,Z =3.28,P ＜ 0.01].The concentration of serum Mg,Ca,Cu and Zn ions in ≥70 years old group was 15.09 (13.64,24.13),68.67 (58.67,97.24),0.97 (0.72,1.34),and 0.54 (0.48,0.74) mg/L,respectively,which was lower than those in ＜ 40 years old group [21.67 (20.08,22.76),98.71 (90.77,113.97),1.35 (1.21,1.71),and 0.78 (0.73,1.01) mg/L],the differences were statistically significant (Z =2.26,2.99,3.01,3.34,P ＜ 0.05).Fe ion in 40-49 years old group was 1.77 (1.45,3.02) mg/L,in 50-59 years old group was 1.92 (1.44,2.66) mg/L,both of them were higher than those of ＜ 40 years old group [1.34 (0.94,1.57) mg/L,Z =-3.25,-2.89,P ＜ 0.05].People whose daily intake of tea fluoride over 4 mg had a lower concentration of Ca ion [98.22 (75.48,111.22) mg/L] than people whose daily intake of tea fluoride ＜ 2 mg [110.24 (97.50,113.97) mg/L,Z =2.41,P ＜ 0.05].No significant difference was found between different degrees of skeletal fluorosis and non-skeletal fluorosis (P ＞ 0.05).Conclusions In the Tibetans who lived in the drinking tea type fluorosis areas,the concentration of serum Mg,A1,Ca,Fe,Zn and Cu ions is similar between skeletal fluorosis and non-skeletal fluorosis,which is also similar between higher daily intake of tea fluoride and lower daily intake of tea fluoride.However,it is different between older people and younger people.

Objective To investigate the effects of excessive aluminum on osteoclastic formation and function in C57BL/6 mice.Methods Six-week-old male C57BL/6 mice (n =20) were randomized by weight and divided into control group and treatment group.The mice were assigned to distilled water and 270 mg/L aluminum ion,respectively.After the 15-week treatment period,the mice were euthanized by ether asphyxiation.Concentration of serum aluminum ion was determined by inductively coupled plasma (ICP).TRAP+ cells in vivo were observed using histomorphometry and osteoclasts microstructure using electron microscope (TEM).Osteoclasts were induced by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor κB ligand (RANKL) with bone marrow macrophages separated from femur and tibia.The expression of mRNA related with bone resorption was detected,including c-FBJ osteosarcoma oncogene (c-Fos),nuclear factor of activated T-cells cl (NFATc1),c-nonreceptor tyrosine kinase (c-Src),dentrocyte expressed seven transmembrane protein (DC-STAMP),d2 isoform of vacuolar (H1) ATPase v0 domain (ATP6v0d2) and matrix metalloproteinase 9 (MMP-9).Results The data showed that the treatment group [(12.04 ± 0.21)mg/L] had significantly increased serum aluminum ion compared with the control group [(11.00 ± 0.04)mg/L,F =10.286,P ＜ 0.05].TRAP+ cells examination confirmed osteoclasts in treatment group [(31.39 ± 9.80) number] were significantly higher than those in the control group [(5.46 ± 4.15) number,F =9.344,P ＜ 0.05].A large proportion of osteoclasts in treatment group lacked ruffled borders and showed vacuolation degeneration.The expression of mRNA in treatment group was lower than that of the control group.The expression of mRNA in control group and treatment group was NFATcl (3.25 ± 0.93 vs.0.29 ± 0.18,F =11.602,P ＜ 0.05),c-Fos (0.86 ± 0.16 vs.0.16 ± 0.02,F =9.405,P ＜ 0.05),c-Src (8.82 ± 1.51 vs.2.29 ± 0.36,F =9.128,P ＜ 0.05),DC-STAMP (3.70 ± 0.70 vs.1.36 ± 0.57,F =10.298,P ＜ 0.05),ATP6v0d2 (15.60 ± 4.81 vs.1.39 ± 0.95,F =8.828,P ＜ 0.05),and MMP-9 (18.64 ± 7.62 vs.2.10 ± 0.92,F =9.356,P ＜ 0.05),respectively.Conclusions Aluminum can increase the number of osteoclasts under epiphyseal plate,but inhibits osteoclasts differentiation.This phenomena may be related with decreased expression of c-Src,DC-STAMP,c-Fos,NFATcl,ATP6v0d2 and MMP-9 mRNA which regulate the function of osteoclasts.