Objective To explore the effect and possible mechanism of berberine on the macro-phage polarization of human myeloid leukemia monocytic cell line THP-1 induced by oxidized low-density lipoprotein(ox-LDL).Methods THP-1 cells were induced into macrophages by PMA,and then according to different concentrations of berberine,the cells were divided into con-trol group,and 5,10,20,40 and 50 μmol/L berberine groups.After intervention for 24 or 48 h,CCK8 assay was used to detect cell viability for optimal concentration and time of berberine treat-ment.PMA-induced THP-1 macrophages were assigned into blank group,model group(ox-LDL),berberine group,inhibitor group(phosphatidyl inositol 3-kinase(PI3K)inhibitor LY294002)and berberine+inhibitor group(berberine+LY294002).The contents of inducible nitric oxide syn-thase(iNOS)and TGF-β1 were detected by ELISA.qPCR was employed to measure the mRNA expression of TNF-α,arginase 1(Arg1),PI3K and protein kinase B Akt1,and Western blotting was applied to detect the protein levels of Akt1 and phosphorylated protein kinase B antibody(p-Akt1).Results In 24 h after intervention,the macrophage activity was significantly lower in the 40 and 50 μmol/L berberine groups than the control group(P＜0.05),and after 48 h,the ac-tivity in all the 5 doses of berberine groups was obviously lower than that in the control group[(0.89±0.02)％,(0.82±0.03)％,(0.71±0.02)％,(0.62±0.03)％and(0.53±0.02)％vs(1.01±0.01)％,P＜0.05].Berberine treatment of 20 μmol/L for 24 h had little effect on cell viability,and the dose and the time were regarded as the best concentration and time.Compared with the blank group,iNOS content and TNF-α mRNA level were increased in the model group,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1,and p-Akt1/Akt1 protein levels were de-creased(P＜0.05).iNOS content and TNF-α mRNA level were decreased,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1s were increased in the berberine group than the model group(P＜0.05).Compared with the berberine group,iNOS con-tent and TNF-α mRNA level were increased,while mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1 were decreased in the berberine+inhibitor group(P＜0.05).Con-clusion Berberine can inhibit the inflammatory response of THP-1 macrophages induced by ox-LDL by activating PI3K/Akt1 pathway,and inhibit the M1 polarization and promote the M2 polarization of macrophages.

Objective:To investigate the correlation between serum recombinant sclerostin (SOST) and dickkopf-related protein 1 (DKK-1) levels and bone metabolism indexes in patients with osteoporosis after differentiated thyroid cancer surgery.Methods:A total of 110 patients diagnosed with osteoporosis after differentiated thyroid cancer surgery were recruited as the study group, and another 110 patients without osteoporosis diagnosed after differentiated thyroid cancer surgery were recruited as the control group. The general data, bone mineral density, serum SOST, DKK-1 levels and bone metabolism indicators N-terminal propeptide of type I procollagen (PINP), bone alkaline phosphatase (BALP), beta-isomerized C-telopeptide (β-CTX), 25-hydroxyvitamin D3 [25- (OH) D3] levels were compared between the two groups. The correlation between serum SOST, DKK-1 levels and bone metabolism indexes was analyzed, and the risk factors affecting the formation of osteoporosis were explored.Results:The T value of bone mineral density in the study group (-3.27±0.92) was significantly lower than that in the control group (-1.23±0.27, t=22.32, P<0.001). The serum SOST (15.84±1.34, t=32.53, P<0.001) and DKK-1 (5.96±1.40, t=4.82, P<0.001) levels in the study group were significantly higher than those in the control group (SOST: 10.24±1.21, DKK-1: 5.05±1.40). The serum PINP (40.95±9.84, t=7.59, P<0.001), BALP (23.14±5.26, t=5.06, P<0.001) and β-CTX (1.07±0.54, t=4.96, P<0.001) in the study group were significantly higher than those in the control group (31.48±8.64, 19.64±4.99, 0.78±0.29), and the 25- (OH) D3 level (13.68±4.49) was significantly lower than that of the control group (18.31±5.72, t=6.68, P<0.001). Serum SOST was positively correlated with PINP ( r=0.33, P=0.001), BALP ( r=0.23, P=0.016) and β-CTX ( r=0.19, P=0.046), but not with 25- (OH) D3 ( r=-0.09, P=0.349). Serum DKK-1 was positively correlated with PINP ( r=0.19, P=0.044), BALP ( r=0.26, P=0.007) and β-CTX ( r=0.21, P=0.028), but not with 25- (OH) D3 ( r=-0.16, P=0.088). Serum SOST and DKK-1 levels were independent risk factors for osteoporosis (all P<0.05) . Conclusion:Serum SOST and DKK-1 levels are independent risk factors for the formation of osteoporosis, which are significantly positively correlated with bone metabolism indexes PINP, BALP, and β-CTX in patients with osteoporosis after differentiated thyroid cancer surgery.

Objective:To investigate the correlation between adiponectin gene polymorphism and osteoporosis in patients with differentiated thyroid after thyroid stimulating hormone (thyroid stimulating hormone, TSH) suppression therapy.Methods:A total of 79 patients who underwent resection of thyroid cancer and TSH suppression therapy were collected as research objects. After 5 years of follow-up, the bone mineral density of the patients was measured, and they were divided into normal bone mass group and osteoporosis group. The general data of the two groups of patients were compared, and the distribution frequencies of rs1063539, rs266729, rs3774261, and rs710445 genotypes in the two groups of patients were analyzed. The differences in bone mineral density of patients with different genotypes of rs1063539, rs266729, rs3774261, and rs710445 were analyzed. To explore the risk factors of osteoporosis in patients treated with TSH suppression after differentiated thyroid surgery.Results:General data analysis showed that the family history of osteoporosis ( P=0.021) and preoperative thyroid hormone status ( P=0.022) were significantly different between the two groups (all P<0.05). The genotype frequency deviations of rs1063539, rs266729, rs3774261, and rs710445 conformed to the Hardy-Weinberg equilibrium law. The distribution of the three genotypes of rs1063539 locus was significantly different between the two groups, and the bone mineral density T value of rs1063539 CC+CG genotype (-3.68±0.61) was significantly lower than that of GG type (-3.14±0.47) ( t=3.142, P=0.003). Logistic regression analysis showed that no family history of osteoporosis was a protective factor for osteoporosis in patients with TSH suppression after thyroid cancer surgery ( OR: 0.258, OR 95%CI: 0.082-0.773, P=0.020). Preoperative hyperthyroidism ( OR: 2.203, OR 95%CI: 1.134-4.541, P=0.025) and rs1063539 CC+CG genotype ( OR: 4.392, OR 95%CI: 1.248-17.652, P=0.027) were the risk factors inducing osteoporosis. Conclusion:Adiponectin rs1063539 gene polymorphism is associated with bone mineral density in patients, and rs1063539 CC+CG genotype can increase the risk of osteoporosis in patients treated with TSH suppression after differentiated thyroid surgery.

Objective:To investigate the expression of miRNA-221-3p (miR-221-3p) in pancreatic cancer cells and its effect on apoptosis of pancreatic cancer cells, and the possible related mechanisms.Methods:Pancreatic cancer cell line PATU8988T was selected and transfected with miR-221-3p mimics, miR-221-3p inhibitors and their corresponding negative control sequences using Lipofectamine 3000. PATU8988T cells were divided into negative control group (without any treatment), miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group, and miR-221-3p inhibitors group. Real-time quantitative fluorescence polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-221-3p, flow cytometry was used to detect the influence of miR-221-3p on apoptosis of pancreatic cancer cells, and Western blotting was used to detect the expressions of P53 and PTEN proteins in PATU8988T cell line.Results:The relative expression levels of miR-221-3p in negative control group, miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group and miR-221-3p inhibitors group were 1.02±0.18, 1.50±0.33, 2.96±0.70, 1.62±0.30, and 0.36±0.05, respectively, and the difference was statistically significant ( F = 12.61, P < 0.05); the relative expression level of miR-221-3p in miR-221-3p mimics group was higher than that in negative control group and miR-221-3p mimics negative control group ( t = 1.94, P < 0.05; t = 1.45, P < 0.05); the relative expression level of miR-221-3p in miR-221-3p inhibitors group was lower than that in negative control group and miR-221-3p inhibitors negative control group ( t = -0.65, P < 0.05; t = -1.26, P < 0.05). The apoptosis rates in negative control group, miR-221-3p mimics negative control group, miR-221-3p mimics group, miR-221-3p inhibitors negative control group and miR-221-3p inhibitors group were (8.60±0.20)%, (8.60±0.26)%, (4.27±0.31)%, (8.83±0.29)%, and (13.63±0.60)%, respectively, and the difference was statistically significant ( F = 253.80, P < 0.01); the apoptosis rates in miR-221-3p mimics group was lower than that in negative control group and miR-221-3p mimics negative control group ( t = -4.33, P < 0.05; t = 4.33, P < 0.05); the apoptosis rate in miR-221-3p inhibitors group was higher than that in negative control group and miR-221-3p inhibitors negative control group ( t = 5.03, P < 0.05; t = 4.80, P < 0.05). There was no statistical difference in expression levels of P53 and PTEN proteins between miR-221-3p mimics negative control group and miR-221-3p inhibitors negative control group (P53: t = 0.22, P > 0.05; PTEN: t = 0.33, P > 0.05); the expression levels of P53 and PTEN proteins in miR-221-3p mimics group were decreased compared with the miR-221-3p mimics negative control group (P53: t = 4.31, P < 0.05; PTEN: t = 8.49, P < 0.05); the expression levels of P53 and PTEN proteins in miR-221-3p inhibitors group were increased compared with the miR-221-3p inhibitors negative control group (P53: t = 5.17, P < 0.05; PTEN: t = 6.21, P < 0.05). Conclusions:miR-221-3p is highly expressed in pancreatic cancer PATU8988T cells, which can inhibit the apoptosis of pancreatic cancer cells. miR-221-3p may regulate the progression of pancreatic cancer through P53 and PTEN.

Objective:To investigate the diagnostic value of circular RNA circ_100367 in thyroid cancer (THCA) and its relationship with immune-related factors.Methods:According to the data chip provided by the National Center for Biotechnology Information (NCBI) website, the differentially expressed circRNAs in THCA were analyzed, then circ_100367 was included in this study. The serum of 175 THCA patients and healthy people were collected, and the expression levels of circ_100367 and its linear transcript DCAF8 mRNA in serum samples were detected by qRT-PCR, and the correlation between circ_100367 and DCAF8 was calculated. The correlation between the expression of circ_100367 and the clinicopathological characteristicsof the patients, immune infiltration level and immunosuppressive factor PD-1 was analyzed.Results:Compared with serum of healthy people (1.00±0.37) , expression level of circ_100367 in serum of THCA patients was significantly increased (1.37±0.41) ( t=8.80, P<0.001) , and there was no significant difference in DCAF8 mRNA expression ( t=1.67, P=0.095) , but circ_100367 was positively correlated with DCAF8 mRNA expression ( r=0.17, P=0.028) . Analysis of expression and clinicopathological characteristics of circ_100367 showed that compared with patients in M0 group (1.26±0.40) , circ_100367 was overexpressed in M1 and Mx patients (1.43±0.40) ( t=2.63, P=0.009) ; compared with N0 patients (1.24±0.36) , circ_100367 was overexpressed in serum of N1 and Nx patients (1.45±0.42) ( t=3.48, P=0.001) ; compared with serum of patients with negative lymph node detection (1.28±0.36) , circ_100367 was overexpressed in serum of positive patients (1.42±0.43) ( t=2.14, P=0.034) ; compared with T1+T2 stage patients (1.30±0.37) , circ_100367 expression was overexpressed in serum ofT3+T4 patients (1.40±0.43) ( t=2.22, P=0.028) . Analysis of the expression and immune infiltration levels of circ_100367 found that highly expressed circ_100367 was associated with CD8+ T cells ( r=0.25, P=0.024) , macrophages ( r=0.22, P=0.038) , CD4+ T cells ( r=0.25, P=0.020) and B cell ( r=0.23, P=0.033) levels. The expression of circ_100367 was also positively correlated with the immunosuppressive factor PD-1 ( r=0.19, P=0.011) . Conclusion:circ_100367 can be used as a marker for the diagnosis of THCA and its expression is strongly correlated with immune-related factors.

Objective To summarize clinicopathologic features of papillary thyroid carcinoma (PTC) coexistent with chronic lymphocytic thyroiditis (CLT) and investigate risk factors for lymph node metastasis.Methods The medical records of 4 264 consecutive papillary thyroid carcinoma patients who received surgical treatment from Oct 2013 to Oct 2015 in Peking Union Medical College Hospital were reviewed.The diagnoses was confirmed by histopathological tests.Univariate analysis was performed to identify specific clinicopathologic features of PTC with CLT.Univariate and multivariate analysis were performed to determine whether each clinicopathologic feature was an independent risk factor for lymph node metastasis.Results In all 4 265 cases,there were 3 059 papillary thyroid microcarcinoma (PTMC) (71.7％),1 010 PTC patients (23.7％) with CLT.909 female patients (90％),624 cases with multifocal lesions (61.8％),422 cases with extra-thyroid extension (41.8％),429 cases with lymph node metastasis (42.5％),and 133 cases with metastatic lymph nodes(LNs) ≥6 (13.2％).The median age was 43 years old and median tumor size was 0.8 cm.Patients with CLT were more females (90.0％ vs.70.2％;P ＜ 0.001),younger median age (43 vs.44 years;P =0.001),and lower incidence of lymph node metastasis (42.5％ vs.50.9％;P ＜0.001).CLT was not associated with tumor size,multifocal lesions,extra-thyroid extension and metastatic LNs≥6 (0.8 cm vs.0.7 cm,61.8％ vs.62.9％,41.8％ vs.42.1％ and 13.2％ vs.14.8％,respectively,all P ＞ 0.05).In multivariate analysis,CLT was an independent protective factor for lymph node metastasis (OR =0.713,95％ CI 0.609-0.835,P ＜0.001).In PTC patients with lymph node metastasis,CLT was not associated with lymph node metastasis number (3 vs.3,P =0.300).Conclusions Chronic lymphocytic thyroiditis was an independent protective factor for papillary thyroid carcinoma patients with lymph node metastasis.But in patients with lymph node metastasis,the metastatic number didn't decrease.

Objectives To evaluate the relationship between body mass index (BMI) and the incidence risk of papillary thyroid microcarcinoma (PTMC).Methods This retrospective study included 1210 PTMC patients who underwent surgery between November 2013 and October 2014 in Peking Union Medical College Hospital,China Academy of Medical Science.A population-based 1∶1 matched case-control study was conducted,and each PTMC patients was matched with one who received thyroid function and ultrasonic to confirm that there was no disease in the thyroid.The clinical profiles of these patients were collected.According to Guidelines for Prevention and Control of Overweight and Obesity in Chinese Adults,all subjects were divided into three groups:underweight (BMI ≤ 18.5 kg/m2),normal(18.5 kg/m2 ＜ BMI ≤ 23.9 kg/m2),overweight (24.0 kg/m2 ＜ BMI ＜ 27.9 kg/m2) and obese group(BMI≥28.0 kg/m2).The relationship between BMI and PTMC incidence risk was analyzed by casecontrol study.Univariate and multivariate logistic regression analysis was applied to analyze the relationship between BMI and PTMC severity.Results The BMI of PTMC patients was significant higher than in normal control [(24.30 ±3.33) kg/m2 vs (23.31 ± 3.50) kg/m2,P ＜ 0.0001].Compared with BMI normal group,the incidence risk of PTMC in underweight group was significantly lower (OR =0.449,95 ％ CI:0.270-0.747),which is higher in overweight and obese group (OR =1.559,95％ CI:1.261-1.928;OR =2.059,95％ CI:1.501-2.823).Histopathological review of 1210 PTMC patients with surgical resection revealed.The proportions of underweight,normal,overweight and obese group of the patients with extrathyroid extension (3.1％,48.0％,36.7％,12.2％) have significant differences with those in the patients whose tumor are limited to the thyroid (0.7％,45.2％,36.0％,18.1％) (P =0.0090).The proportions of 4 group of the patients with multiple lesions (3.2％,49.0％,35.6％,12.2％) were significantly differences to those in the patients with single lesion (0.8％,43.3％,38.7％,17.2％) (P =0.0050).Multivariate analysis showed that underweight is a protective factor of extrathyroidal extension (OR =0.219,95 ％ CI:0.051-0.932;OR =0.279,95 ％ CI:0.085-0.935) and mulifocality,and obese is an independent risk factors(OR =1.556,95％CI:1.047-2.312;OR =1.764,95％CI:1.204-2.584).Conclusions This study identified that the incidence risk of PTMC is positive related with BMI.In PTMC patients,obesity increases the risk of mulifocality and extrathyroidal extension.Attention should be paid to the effect of obesity on the incidence risk of PTMC and the diagnosis and treatment in clinical practice.

Objective To evaluate the clinical and pathological feature,as well as risk factors of lymph node metastasis (LNM) and high-volume LNM (hvLNM) in papillary thyroid microcarcinoma (PTMC) with di ameter ≤0.5 cm.Methods PTMC patients who received surgical treatments in Peking Union Medical College Hospital from Nov.2013 to Nov.2014 were reviewed.Patients were allocated into the ≤0.5 cm group and (0.5-1)cm group according to tumor diameter.Clinical and pathological features were assessed and compared.Risk factors of LNM and hvLNM were also assessed through univariate and multivariate analysis.Results 1414 patients were enrolled,of which 315 patients (22.3％) were in the ≤0.5 cm group.76 LNM (24.1％) and 9 hvLNM (2.9％) were detected in the ≤0.5 cm group.There was significantly less capsule invasion (14.3％ vs 25.0％,P＜0.05),LNM (24.1％ vs 39.8％,P＜0.05) and hvLNM (2.9％ vs 7.9％,P＜0.05) in ≤0.5 cm group than in (0.5-1)cm group.In univariate analysis,patients aging ＜40 years old were more likely to have LNM than those older than 40(38.0％ vs 20.1％,P＜0.05),while male patients tended to have more LNM than female (32.4％ vs 21.9％,P=0.073).No risk factors were identified for hvLNM.In multivariate analysis,multifocality and younger than 40 years old were the independent risk factors of LNM (OR=2.082 and 2.899,P＜0.05),while male tended to be the independent risk factors of LNM (OR=l.807,P=0.058).No independent risk factors was identified for hvLNM.Conclusions A certain proportion of PTMC patients are with tumor diameter ≤0.5 cm,who have lower risk of LNM and hvLNM.Dynamic observation may be an option,especially in older ≥40 years old),unifocal and female patients.

Objective: To investigate the related factors for lymph node metastasis (LNM), especially for high volume LNM (>5 metastatic lymph nodes) in papillary thyroid carcinoma (PTC). Methods: The medical records of 2 073 consecutive PTC patients who underwent lobectomy, near-total thyroidectomy or total thyroidectomy with ipsilateral or bilateral central lymph node dissection in Department of General Surgery, Peking Union Medical College Hospital from November 2013 to October 2014 were reviewed. Clinical and pathological features were collected. Univariate and multivariate analysis were performed to identify the related factors for LNM/high volume LNM. Results: In all 2 073 patients, LNM and high volume LNM were confirmed in 936 (45.15%) cases and 254 (12.25%) cases respectively. In univariate analysis, large tumor size, young patients (<40 years), male were associated with both LNM and high volume LNM. In multivariate analysis, tumor size >2.0 cm, young patients (<40 years), male were independent related factors of LNM (OR=5.262, 95% CI: 3.468 to 7.986; OR=2.447, 95% CI: 2.000 to 2.995; OR=1.988, 95% CI: 1.593 to 2.480, respectively, all P=0.000) and high volume LNM (OR=6.687, 95% CI: 4.477 to 9.986; OR=2.975, 95% CI: 2.224 to 3.980; OR=2.354, 95% CI: 1.737 to 3.191, respectively, all P=0.000). In 1 414 PTMC patients, a similar result was also demonstrated.Compared with young patients (<40 years), old patients (≥60 years) had lower incidence of LNM (25.47% vs. 52.24%, χ²=62.903, P=0.000) and high volume LNM (1.89% vs. 13.18%, χ²=37.341, P=0.000). Additionally, old patients also had lower risk of both LNM (OR=0.316, 95% CI: 0.194 to 0.517, P=0.000) and high volume LNM (OR=0.142, 95% CI: 0.034 to 0.599, P=0.000). Conclusions The tumor size was the main related factor for both LNM and high volume LNM in PTC. The treatment should be more active in patients with tumor size >2 cm with consideration of higher incidence and risk for LNM and high volume LNM. Young patient was another important related factor for LNM and high volume LNM. In PTMC, old patients had lower incidence and risk for both LNM and high volume LNM. Dynamic observation or less surgical extent could be an option for these patients.