ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

Objective This study aimed to identify differentially methylated genes (DMGs) associated with natural killer cells in patients with autoimmune thyroiditis (AIT),focusing on the influence of varying water iodine exposure levels. Methods Participants were divided into categories based on median water iodine (MWI) concentrations:iodine-fortified areas (IFA,MWI<10 μg/L),iodine-adequate areas (IAA,40 ≤ MWI ≤ 100μg/L),and iodine-excessive areas (IEA,MWI>300 μg/L). A total of 176 matched AIT cases and controls were recruited and divided into 89,40,and 47 pairs for IFA,IAA,and IEA,respectively. DMGs were identified using 850K BeadChip analysis for 10/10 paired samples. Validation of DNA methylation and mRNA expression levels of the DMGs was conducted using MethylTarget? and QRT-PCR for 176/176 paired samples. Results KLRC1,KLRC3,and SH2D1B were identified as significant DMGs. Validation revealed that KLRC1 was hypomethylated and highly expressed,whereas KLRC3 was hypermethylated and highly expressed in individuals with AIT. Furthermore,KLRC1 was hypomethylated and highly expressed in both IFA and IEA. Conclusion The DNA methylation status of KLRC1 and KLRC3 may play crucial roles in AIT pathogenesis. Additionally,DNA methylation of KLRC1 seems to be influenced by different iodine concentrations in water.

Mice ; Animals ; Spiral Ganglion/physiology* ; Cochlea/physiology* ; Neurons

Objective:To clarify the relationship between interleukin (IL)-6, IL-10 gene polymorphisms and autoimmune thyroid disease (AITD).Methods:Literature search was conducted through databases such as PubMed, Web of Science, CNKI, Embase, Wanfang Database and VIP.com, and domestic and foreign literatures related to IL-6, IL-10 gene polymorphisms and AITD were included in the study. The time limit was from the self-built of the databases to July 2021. Meta-analysis was performed with STATA 16.0 software, the odds ratio ( OR) and 95% confidence interval ( CI) were used as effect indicators, random-effect or fixed-effect model was selected according to the heterogeneity results, and the source of heterogeneity was explored through subgroup analysis. Publication bias was assessed using funnel plots and Egger's test. Results:Finally, 19 literatures were included, all in English. There were 12 studies on IL-6 genes and 11 studies on IL-10 genes, including 4 studies on both IL-6 and IL-10 genes. In the whole population, the loci associated with AITD were IL-6 -174 G/C site (GG vs CC + GC: OR =1.94, 95% CI = 1.01 - 3.76), IL-6 -572 G/C site (GG + GC vs CC: OR = 0.49, 95% CI = 0.29 - 0.84; GG vs CC + GC: OR = 0.76, 95% CI = 0.60 - 0.96; GG + CC vs GC: OR = 0.63, 95% CI = 0.49 - 0.81), IL-10 -819 T/C site (TT + TC vs CC: OR = 1.84, 95% CI = 1.01 - 3.34; T vs C: OR = 1.59, 95% CI = 1.00 - 2.51), and IL-10 -1 082 A/G site (AA + AG vs GG: OR = 0.77, 95% CI = 0.64 - 0.92; AA vs GG + AG: OR = 2.03, 95% CI = 1.16 - 3.58; A vs G: OR = 0.76, 95% CI = 0.61 - 0.94). The results of subgroup analysis showed that in Asian population, the loci associated with AITD were IL-6 -174 G/C site (GG vs CC + GC: OR = 4.61, 95% CI = 1.11 - 19.23; G vs C: OR = 0.65, 95% CI = 0.44 - 0.97); IL-6 -572 G/C site (GG vs CC + GC: OR = 0.64, 95% CI = 0.41 - 0.99; GG + CC vs GC: OR = 0.60, 95% CI = 0.38 - 0.94); IL-10 -819 T/C site (TT + TC vs CC: OR = 2.51, 95% CI = 1.48 - 4.25; T vs C: OR = 1.91, 95% CI = 1.05 - 3.46); and IL-10 -1 082 A/G site (AA + AG vs GG: OR = 0.66, 95% CI = 0.52 - 0.84; AA vs GG + AG: OR = 2.83, 95% CI = 1.54 - 5.21; A vs G: OR = 0.66, 95% CI = 0.53 - 0.82). Conclusion:IL-6 -174 G/C, IL-6 -572 G/C, IL-10 -819 T/C and IL-10 -1 082 A/G polymorphisms are associated with the susceptibility to AITD, especially in Asians.

Objective:To verify the determination method of iodine in serum by inductively coupled plasma mass spectrometry (ICP-MS) and to evaluate the consistency between ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry in determination of serum iodine. Methods:Serum iodine concentration was determined by ICP-MS, 187Re was used as an internal standard, and ralated parameters were optimized. Eighty-eight serum samples were simultaneously determined by ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry, and the evaluation indexes included determination range of standard curve, detection limit, precision, accuracy. In addition, we also evaluated the consistency of the two methods through inter-group correlation analysis, intra-group correlation coefficient analysis, Passing-Bablok regression and Bland-Altman analysis. Results:The linear range of ICP-MS standard curve was 0 - 300 μg/L. There was a good linear correlation between iodine concentration value and iodine response value, and the correlation coefficient range was 0.999 8 to 0.999 9. The detection limit of the ICP-MS method was 1.96 μg/L. The relative standard deviation ( RSD) ranged from 0.2% to 1.4% and from 0.4% to 1.8% for intra and inter-batch precision tests of serum samples. The recovery rate ranged from 90.44% to 108.71%. The correlation analysis of 88 serum samples showed that there was a good correlation between the two methods ( r = 0.934, P < 0.05), and the intra-class correlation coefficient was 0.932. The results of Passing-Bablok regression showed that there was no significant difference between the two methods ( P > 0.05). Bland-Altman diagram suggested that the results of the two methods were consistent. Conclusions:ICP-MS method has low detection limit, high precision and accuracy. ICP-MS method is simple, rapid, easy and suitable for determination of iodine in large quantities of serum samples. The results of the two methods for determining serum iodine are consistent.

Objective: To explore the application value of MRI quantitative analysis in evaluation on therapeutic effect of rheumatoid arthritis (RA) of the wrist. Methods: Totally 30 patients with RA after regular treatment for 1 year were enrolled. The wrist joint was scanned before and after treatment, and the volume of synovitis and bone marrow edema before and after treatment were measured by using the quantitative analysis software. The erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity score of 28 (DAS28) were also obtained. Changes of synovitis, bone marrow edema, CRP, ESR and DAS28 were analyzed before and after treatment, correlation analysis was performed on the changes of synovitis and bone marrow edema with changes of CRP, ESR and DAS28. Results: After treatment, the volume of synovitis, the range of bone marrow edema, CRP, ESR and DAS28 of the patients reduced significantly (all P<0.05). Synovitis was strongly correlated with changes in bone marrow edema (r=0.61, P<0.01). Changes in synovitis and bone marrow edema were not significantly correlated with those of ESR and CRP (all P>0.05), while changes in synovitis and bone marrow edema were moderately correlated with changes of DAS28 (r=0.50, 0.56, P=0.01,<0.01). Conclusion: MRI quantitative analysis can be used as accurate and Objective: quantitative indices for monitoring and evaluating therapeutic effect of RA.

Huntington's disease (HD) is a deadly neurodegenerative disease with abnormal expansion of CAG repeats in the huntingtin gene. Mutant Huntingtin protein (mHTT) forms abnormal aggregates and intranuclear inclusions in specific neurons, resulting in cell death. Here, we tested the ability of a natural heat-shock protein 90 inhibitor, Gedunin, to degrade transfected mHTT in Neuro-2a cells and endogenous mHTT aggregates and intranuclear inclusions in both fibroblasts from HD patients and neurons derived from induced pluripotent stem cells from patients. Our data showed that Gedunin treatment degraded transfected mHTT in Neuro-2a cells, endogenous mHTT aggregates and intranuclear inclusions in fibroblasts from HD patients, and in neurons derived from induced pluripotent stem cells from patients in a dose- and time-dependent manner, and its activity depended on the proteasomal pathway rather than the autophagy route. These findings also showed that although Gedunin degraded abnormal mHTT aggregates and intranuclear inclusions in cells from HD patient, it did not affect normal cells, thus providing a new perspective for using Gedunin to treat HD.

Objective: To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging (DKI) technique, and to analyze the correlation with clinical manifestations. Methods: Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classified into gait disorder subtype (PIGD, n=57) and tremor dominant subtype (TD, n=40)subtypes according to motor symptoms. Fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (Da), radial diffusivity(Dr), mean kurtosis (MK), axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were generated by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres, including red nucleus(RN), substantia nigra pars reticulate(SNr), substantia nigra pars compacta(SNc), putamen(PUT), globus pallidus(GP), head of caudate nucleus (CN)and thalamus(THA). Results: TD showed a higher MMSE score(P=0.019), but lower modified Hoehn-Yahr score than that in PIGD (P<0.001), there was no significant difference of age of onset, sex, limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD, Da and Dr and up-regulated Ka values(P<0.001); MK(0.83±0.26, 0.80±0.18) was increased in SNr both in TD and PIGD, while SNc, PUT and GP (0.84±0.20, 0.75±0.07, 0.81±0.14)were decreased only in TD (P=0.017, P=0.010, P=0.020, P<0.001, P=0.002). The Kr values of PUT and CN(0.71±0.17, 0.72±0.14) were reduced in PIGD, while CN(0.70±0.14) were reduced in TD respectively (P=0.002, P=0.031, P=0.007). The MK was lower in TD than that in PIGD (t=-2.214, P=0.029), and no significant difference was found in other grey matter nuclei between TD and PIGD(P>0.05). Moreover, there was no significant correlation between DKI value and disease duration, MMSE score or Hoehn-Yahr scale (P>0.05) in TD and PIGD. Conclusion There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.

Objective To investigate the difference in the microstructure of gray matter nucleus in different movement subtypes of Parkinson’s disease (PD) by diffusion kurtosis imaging ( DKI) technique, and to analyze the correlation with clinical manifestations. Methods Ninety-seven patients with PD and 83 healthy controls performed conventional MRI sequence and DKI sequence scan. The PD patients were classi-fied into gait disorder subtype (PIGD,n=57) and tremor dominant subtype (TD,n=40)subtypes according to motor symptoms. Fractional anisotropy (FA),mean diffusivity (MD),axial diffusivity (Da),radial diffu-sivity(Dr),mean kurtosis (MK),axial kurtosis (Ka) and radial kurtosis (Dr) maps and data were genera-ted by software after processing. DKI was performed for all subjects and data was collected from different brain regions in both hemispheres,including red nucleus(RN),substantia nigra pars reticulate( SNr),sub-stantia nigra pars compacta(SNc),putamen(PUT),globus pallidus(GP),head of caudate nucleus (CN)and thalamus(THA). Results TD showed a higher MMSE score(P=0. 019),but lower modified Hoehn-Yahr score than that in PIGD (P<0. 001),there was no significant difference of age of onset,sex,limbs of onset or disease duration between two PD subgroups. Compared with healthy controls, both TD and PIGD showed down-regulated MD,Da and Dr and up-regulated Ka values(P<0. 001); MK(0. 83±0. 26,0. 80±0. 18) was increased in SNr both in TD and PIGD,while SNc,PUT and GP (0. 84± 0. 20,0. 75± 0. 07,0. 81± 0. 14) were decreased only in TD (P=0. 017,P=0. 010,P=0. 020,P<0. 001,P=0. 002). The Kr values of PUT and CN(0. 71±0. 17,0. 72±0. 14) were reduced in PIGD,while CN(0. 70±0. 14) were reduced in TD re-spectively (P=0. 002,P=0. 031,P=0. 007). The MK was lower in TD than that in PIGD (t=-2. 214,P=0. 029),and no significant difference was found in other grey matter nuclei between TD and PIGD ( P>0. 05). Moreover,there was no significant correlation between DKI value and disease duration,MMSE score or Hoehn-Yahr scale (P>0. 05) in TD and PIGD. Conclusion There is heterogeneity of clinical symptoms between these two subgroups of PD. DKI can quantify the microstructural changes of grey matter nucleus in different type PD patient.