The activator protein-1 complex governs a vascular degenerative transcriptional programme in smooth muscle cells to trigger aortic dissection and rupture.

BACKGROUND AND AIMS: Stanford type A aortic dissection (AD) is a degenerative aortic remodelling disease marked by an exceedingly high mortality without effective pharmacologic therapies. Smooth muscle cells (SMCs) lining tunica media adopt a range of states, and their transformation from contractile to synthetic phenotypes fundamentally triggers AD. However, the underlying pathomechanisms governing this population shift and subsequent AD, particularly at distinct disease temporal stages, remain elusive. METHODS: Ascending aortas from nine patients undergoing ascending aorta replacement and five individuals undergoing heart transplantation were subjected to single-cell RNA sequencing. The pathogenic targets governing the phenotypic switch of SMCs were identified by trajectory inference, functional scoring, single-cell regulatory network inference and clustering, regulon, and interactome analyses and confirmed using human ascending aortas, primary SMCs, and a ß-aminopropionitrile monofumarate-induced AD model. RESULTS: The transcriptional profiles of 93 397 cells revealed a dynamic temporal-specific phenotypic transition and marked elevation of the activator protein-1 (AP-1) complex, actively enabling synthetic SMC expansion. Mechanistically, tumour necrosis factor signalling enhanced AP-1 transcriptional activity by dampening mitochondrial oxidative phosphorylation (OXPHOS). Targeting this axis with the OXPHOS enhancer coenzyme Q10 or AP-1-specific inhibitor T-5224 impedes phenotypic transition and aortic degeneration while improving survival by 42.88% (58.3%-83.3% for coenzyme Q10 treatment), 150.15% (33.3%-83.3% for 2-week T-5224), and 175.38% (33.3%-91.7% for 3-week T-5224) in the ß-aminopropionitrile monofumarate-induced AD model. CONCLUSIONS: This cross-sectional compendium of cellular atlas of human ascending aortas during AD progression provides previously unappreciated insights into a transcriptional programme permitting aortic degeneration, highlighting a translational proof of concept for an anti-remodelling intervention as an attractive strategy to manage temporal-specific AD by modulating the tumour necrosis factor-OXPHOS-AP-1 axis.

Human regulatory macrophages are potent in suppression of the xenoimmune response via indoleamine-2,3-dioxygenase-involved mechanism(s).

BACKGROUND: For xenotransplantation to truly succeed, we must develop immunomodulatory strategies to suppress the xenoimmune response but by minimizing immunosuppression over the long term. Regulatory macrophages (Mreg) have been shown to suppress polyclonal T-cell proliferation in vitro and prolong allograft survival in vivo. However, the question of whether they are capable of suppressing xenoimmune responses remains unknown. This study assessed the potential of human Mreg to be used as an effective immunomodulatory method in xenotransplantation. METHODS: CD14+ monocytes selected from human peripheral blood mononuclear cells (PBMC) were cultured with macrophage colony-stimulating factor (M-CSF) for 7 days with IFN-γ added at day 6 for Mreg induction. Mreg phenotyping was performed by flow cytometric analysis, and the in vitro suppressive function was assessed by mixed lymphocyte reaction (MLR) using irradiated pig PBMC as the xenogeneic stimulator cells, human PBMC as responder cells, and autologous Mreg as suppressor cells. To assess mRNA expression of Mreg functional molecules indoleamine-2,3-dioxygenase (IDO), IL-10, inducible nitric oxide synthase (iNOS) and TGF-ß were measured by real-time PCR. Supernatants were collected from the MLR cultures for IDO activity assay by high-performance liquid chromatography (HPLC). The effects of the IDO inhibitor 1-D/L-methyl-tryptophan (1-MT), iNOS inhibitor NG -monomethyl-l-arginine (L-NMMA), and anti-IFN-γ or anti-TGF-ß monoclonal antibody (mAb) treatment on Mreg suppressive capacity were tested from the supernatants of the MLR assays. RESULTS: We demonstrated that induced Mreg with a phenotype of CD14low CD16-/low CD80low CD83-/low CD86+/hi HLA-DR+/hi were capable of suppressing proliferating human PBMC, CD4+, and CD8+ T cells, even at a higher responder:Mreg ratio of 32:1 in a pig-human xenogeneic MLR. The strong suppressive potency of Mreg was further correlated with their upregulated IDO expression and activity. The IDO upregulation of Mreg was associated with an increased production of IFN-γ, an IDO stimulator, by xenoreactive responder cells in the xenogeneic MLR. While no effect on Mreg suppressive potency was detected by addition of the iNOS inhibitor L-NMMA or anti-TGF-ß mAb into the MLR assays, inhibition of IDO activity by neutralizing IFN-γ or by IDO inhibitor 1-MT substantially impaired the capacity of Mreg to suppress the xenogeneic response, indicating the importance of upregulated IDO activity in Mreg-mediated suppression of the xenogeneic response in vitro. CONCLUSION: This study demonstrates that human Mreg are capable of suppressing the xenoimmune response in vitro via IDO-involved mechanism(s), suggesting their potential role as an effective immunomodulatory tool in xenotransplantation.

Dietary high-fat lard intake induces thyroid dysfunction and abnormal morphology in rats.

AIM: Excess dietary fat intake can induce lipotoxicity in non-adipose tissues. The aim of this study was to observe the effects of dietary high-fat lard intake on thyroid in rats. METHODS: Male Sprague-Dawley rats were fed a high-fat lard diet for 24 weeks, and then the rats were fed a normal control diet (acute dietary modification) or the high-fat lard diet for another 6 weeks. The serum lipid profile, total thyroxine (TT4), free thyroxine (FT4) and thyrotropin (TSH) levels were determined at the 12, 18, 24 and 30 weeks. High-frequency ultrasound scanning of the thyroid glands was performed at the 24 or 30 weeks. After the rats were sacrificed, the thyroid glands were collected for histological and immunohistochemical analyses. RESULTS: The high-fat lard diet significantly increased triglyceride levels in both the serum and thyroid, and decreased serum TT4 and FT4 levels in parallel with elevated serum TSH levels. Ultrasonic imaging revealed enlarged thyroid glands with lowered echotexture and relatively heterogeneous features in the high-fat lard fed rats. The thyroid glands from the high-fat lard fed rats exhibited enlarged follicle cavities and flattened follicular epithelial cells under light microscopy, and dilated endoplasmic reticulum cisternae, twisted nuclei, fewer microvilli and secretory vesicles under transmission electron microscopy. Furthermore, the thyroid glands from the high-fat lard fed rats showed markedly low levels of thyroid hormone synthesis-related proteins TTF-1 and NIS. Acute dietary modification by withdrawal of the high-fat lard diet for 6 weeks failed to ameliorate the high-fat lard diet-induced thyroid changes. CONCLUSION: Dietary high-fat lard intake induces significant thyroid dysfunction and abnormal morphology in rats, which can not be corrected by short-term dietary modification.

Hepatocyte miR-21-5p-deficiency alleviates APAP-induced liver injury by inducing PPARγ and autophagy.

Acetaminophen (APAP)-induced liver injury is one of the most frequent causes of acute liver failure worldwide. Significant increases in the levels of miRNA-21 in both liver tissues and plasma have been observed in APAP-overdosed animals and humans. However, the mechanistic effect of miRNA-21 on acute liver injury remains unknown. In this study, we generated a new hepatocyte-specific miRNA-21 knockout (miR-21-HKO) mouse line. miR-21-HKO and the background-matched sibling wild-type (WT) mice were treated with a toxic dose of APAP. Compared with WT mice, miR-21 HKO mice showed an increased survival, a reduction of necrotic hepatocytes, and an increased expression of light chain 3 beta, which suggested an autophagy activation. The expression of PPARγ was highly induced in the livers of miR-21-HKO mice after a 2-h APAP treatment, which preceded the activation of LC3B at the 12 h APAP treatment. miR-21 negatively regulated PPARγ protein expression by targeting its 3'-UTR. When PPARγ function was blocked by a potent antagonist GW9662 in miR-21-HKO mice, the autophage activation was significantly diminished, suggesting an indispensable role of PPARγ signaling pathway in miR-21-mediated hepatotoxicity. Taken together, hepatocyte-specific depletion of miRNA-21 alleviated APAP-induced hepatotoxicity by activating PPARγ and autophagy, demonstrating a crucial new regulatory role of miR-21 in APAP-mediated liver injury.

Intragraft memory-like CD127hiCD4+Foxp3+ Tregs maintain transplant tolerance.

CD4+Foxp3+ regulatory T cells (Tregs) play an essential role in suppressing transplant rejection, but their role within the graft and heterogeneity in tolerance are poorly understood. Here, we compared phenotypic and transcriptomic characteristics of Treg populations within lymphoid organs and grafts in an islet xenotransplant model of tolerance. We showed Tregs were essential for tolerance induction and maintenance. Tregs demonstrated heterogeneity within the graft and lymphoid organs of tolerant mice. A subpopulation of CD127hi Tregs with memory features were found in lymphoid organs, presented in high proportions within long-surviving islet grafts, and had a transcriptomic and phenotypic profile similar to tissue Tregs. Importantly, these memory-like CD127hi Tregs were better able to prevent rejection by effector T cells, after adoptive transfer into secondary Rag-/- hosts, than naive Tregs or unselected Tregs from tolerant mice. Administration of IL-7 to the CD127hi Treg subset was associated with a strong activation of phosphorylation of STAT5. We proposed that memory-like CD127hi Tregs developed within the draining lymph node and underwent further genetic reprogramming within the graft toward a phenotype that had shared characteristics with other tissue or tumor Tregs. These findings suggested that engineering Tregs with these characteristics either in vivo or for adoptive transfer could enhance transplant tolerance.

Association between metabolic obesity phenotypes and multiple myeloma hospitalization burden: A national retrospective study.

Background & purpose: Obesity and metabolic disorders were associated with increased risk of MM, a disease characterized by high risk of relapsing and require frequent hospitalizations. In this study, we conducted a retrospective cohort study to explore the association of metabolic obesity phenotypes with the readmission risk of MM. Patients & methods: We analyzed 34,852 patients diagnosed with MM from the Nationwide Readmissions Database (NRD), a nationally representative database from US. Hospitalization diagnosis of patients were obtained using ICD-10 diagnosis codes. According to obesity and metabolic status, the population was divided into four phenotypes: metabolically healthy non-obese (MHNO), metabolically unhealthy non-obese (MUNO), metabolically healthy obese (MHO), and metabolically unhealthy obese (MUO). The patients with different phenotypes were observed for hospital readmission at days 30-day, 60-day, 90-day and 180-day. Multivariate cox regression model was used to estimate the relationship between obesity metabolic phenotypes and readmissions risk. Results: There were 5,400 (15.5%), 7,255 (22.4%), 8,025 (27.0%) and 7,839 (35.6%) unplanned readmissions within 30-day, 60-day, 90-day and 180-day follow-up, respectively. For 90-day and 180-day follow-up, compared with patients with the MHNO phenotype, those with metabolic unhealthy phenotypes MUNO (90-day: P = 0.004; 180-day: P = < 0.001) and MUO (90-day: P = 0.049; 180-day: P = 0.004) showed higher risk of readmission, while patients with only obesity phenotypes MHO (90-day: P = 0.170; 180-day: P = 0.090) experienced no higher risk. However, similar associations were not observed for 30-day and 60-day. Further analysis in 90-day follow-up revealed that, readmission risk elevated with the increase of the combined factor numbers, with aHR of 1.068 (CI: 1.002-1.137, P = 0.043, with one metabolic risk factor), 1.109 (CI: 1.038-1.184, P = 0.002, with two metabolic risk factors) and 1.125 (95% CI: 1.04-1.216, P = 0.003, with three metabolic risk factors), respectively. Conclusion: Metabolic disorders, rather than obesity, were independently associated with higher readmission risk in patients with MM, whereas the risk elevated with the increase of the number of combined metabolic factors. However, the effect of metabolic disorders on MM readmission seems to be time-dependent. For MM patient combined with metabolic disorders, more attention should be paid to advance directives to reduce readmission rate and hospitalization burden.

Human HLA-DR+CD27+ regulatory T cells show enhanced antigen-specific suppressive function.

Regulatory T cells (Tregs) have potential for the treatment of autoimmune diseases and graft rejection. Antigen specificity and functional stability are considered critical for their therapeutic efficacy. In this study, expansion of human Tregs in the presence of porcine PBMCs (xenoantigen-expanded Tregs, Xn-Treg) allowed the selection of a distinct Treg subset, coexpressing the activation/memory surface markers HLA-DR and CD27 with enhanced proportion of FOXP3+Helios+ Tregs. Compared with their unsorted and HLA-DR+CD27+ double-positive (DP) cell-depleted Xn-Treg counterparts, HLA-DR+CD27+ DP-enriched Xn-Tregs expressed upregulated Treg function markers CD95 and ICOS with enhanced suppression of xenogeneic but not polyclonal mixed lymphocyte reaction. They also had less Treg-specific demethylation in the region of FOXP3 and were more resistant to conversion to effector cells under inflammatory conditions. Adoptive transfer of porcine islet recipient NOD/SCID IL2 receptor γ-/- mice with HLA-DR+CD27+ DP-enriched Xn-Tregs in a humanized mouse model inhibited porcine islet graft rejection mediated by 25-fold more human effector cells. The prolonged graft survival was associated with enhanced accumulation of FOXP3+ Tregs and upregulated expression of Treg functional genes, IL10 and cytotoxic T lymphocyte antigen 4, but downregulated expression of effector Th1, Th2, and Th17 cytokine genes, within surviving grafts. Collectively, human HLA-DR+CD27+ DP-enriched Xn-Tregs expressed a specific regulatory signature that enabled identification and isolation of antigen-specific and functionally stable Tregs with potential as a Treg-based therapy.

Evolution and major changes of the diagnosis and treatment protocol for COVID-19 patients in China 2020-2023.

Since the identification of the first case of pneumonia of unknown cause in 2019, the COVID-19 pandemic has spread the globe for over 3 years. As the most populous country in the world, China's disease prevention policies and response plans concern the health of the country's 1.4 billion people and beyond. During the course of the pandemic, scientific research has been accumulated and given evidence-based support to the official guidance of COVID-19 management. The National Health Commission of China have compiled, published, and updated a total of 10 versions of the "Diagnosis and Treatment Protocol for COVID-19 Patients" to better inform clinical practitioners and staff to effectively screen, diagnose, manage, treat, and care for cases of severe acute respiratory syndrome coronavirus 2 infection. This paper compares and summarizes each version of the protocol in terms of etiology and epidemiology, clinical manifestation and diagnosis, treatment and nursing, disease control and management, presenting detailed changes, additions, deletions, and refinement of the protocols.

TAZ Is Related to Postoperative In-Hospital Mortality of Acute Type A Aortic Dissection.

Background: Surgical repair of acute type A aortic dissection (ATAAD) has high risk and mortality, and there are few biomarkers of postoperative in-hospital mortality until now. This study investigated the association between WW domain-containing transcription regulator protein 1 (TAZ) and the postoperative in-hospital mortality of ATAAD patients. Methods: This is a retrospective cohort study. Data and blood samples were collected from 95 consecutive patients with ATAAD who underwent surgeries in our hospital from July 1, 2016, to December 31, 2016. The data collection included all the risk factors introduced by the modified EuroSCORE (European System for Cardiac Operative Risk Evaluation). The predictors of postoperative in-hospital death were confirmed by univariate regression analysis. Multivariable logistic regressions were used to analyze the association of the preoperative plasma level of TAZ and the postoperative in-hospital mortality of ATAAD patients. In addition, we used the generalized additive model to identify non-linear relationships. Results: Three models were used in the multivariable logistic regression analysis of the relationship between the preoperative plasma level of TAZ and postoperative in-hospital death. In the crude model, the preoperative plasma level of TAZ showed a positive correlation with postoperative in-hospital death [odds ratio (OR) = 1.33, 95% confidence interval (CI): 1.01-1.74, P = 0.04]. In adjusted model I and adjusted model II, similar results were found (OR = 1.35, 95% CI: 1.01-1.80, P = 0.04 and OR = 1.35, 95% CI: 1.01-1.81, P = 0.04). The risk of postoperative in-hospital death in the preoperative plasma level of the TAZ≥12.70 ng/mL group was 10.08 times (OR = 10.08, 95% CI: 1.63-62.37; P = 0.01) that of the preoperative plasma level of the TAZ <12.70 ng/mL group. Conclusions: The high preoperative plasma level of TAZ suggested poor surgical prognosis for ATAAD patients. The patients with a preoperative plasma level of TAZ ≥ 12.7 ng/ml had much higher postoperative in-hospital mortality.

Thyroid Hormone Is Related to Postoperative AKI in Acute Type A Aortic Dissection.

Background: Renal function is profoundly influenced by thyroid hormone levels. This study was designed to evaluate the association between preoperative thyroid hormones and postoperative acute kidney injury (AKI) in acute type A aortic dissection (ATAAD) patients. Methods: A total of 88 patients with ATAAD who underwent surgeries in Beijing Anzhen Hospital and 274 healthy controls from July 2016 to December 2016 were included in this study. Propensity-score matching was used to compare thyroid hormone levels. Additionally, in a cohort study of ATAAD patients, multivariable regression and stratification analyses were conducted to examine the association of preoperative thyroid hormones with postoperative AKI. Results: Compared with healthy controls, ATAAD patients presented with lower preoperative levels of total triiodothyronine (TT3) (P < 0.01), free triiodothyronine (FT3) (P < 0.01), and thyroid-stimulating hormone (TSH) (P < 0.01) and a higher preoperative level of free thyroxine (FT4) (P < 0.01). The overall occurrence of postoperative AKI was 45.5%. Multivariate regression revealed that low levels of TT3 (OR = 0.07, 95% CI, 0.01-0.86, P = 0.04) were independently associated with postoperative AKI. Subgroup analyses showed that the association between TT3 and AKI was significant in patients with normal TSH levels (OR = 0.001 95% CI, 0.001-0.16, P < 0.01) but not in patients with lower TSH levels (P = 0.12). Conclusion: The present study showed that a low level of TT3 was a predictor of postoperative AKI in ATAAD patients, especially in patients with normal TSH. The thyroid function should be checked before surgical intervention of patients with ATAAD, and patients with low T3 might be at higher risk of postoperative AKI.

Fabrication of a Novel Nanofiltration Membrane with Enhanced Performance via Interfacial Polymerization through the Incorporation of a New Zwitterionic Diamine Monomer.

It is known that the polyamide (PA) barrier layer's inherent microstructure and surface physicochemical properties of thin film composite nanofiltration membrane are crucial for its separation performance. Herein, we designed and synthesized a new zwitterionic aromatic diamine monomer 3-(4-(2-((4-aminophenyl)amino)ethyl)morpholino-4-ium)propane-1-sulfonate (PPD-MEPS) through a three steps reaction, and this hydrophilic molecule was incorporated into the active layer to tailor the poly(piperazine-amide)-based nanofiltration membranes with significantly improved water permeability and antifouling properties. As a p-phenylenediamine (PPD) derivative, PPD-MEPS possesses two active amine units, which can react with trimesoyl chloride in the organic phase during the interfacial polymerization reaction process. Thus, the super-hydrophilic zwitterions were not only on the membrane surface but also across the whole PA layer to facilitate water molecule transportation. The successful augmentation of zwitterions into the PA layer was well illustrated by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) results and X-ray photoelectron spectroscopy analysis. With increasing loading content of PPD-MEPS in PIP aqueous solution, the as-fabricated nanofiltration membranes (NFMs) exhibited higher hydrophilicity, increased active layer thickness, and molecular weight cut off. When the zwitterionic monomer reached 60% to PIP for NFM-4, the water permeability went up to 9.82 L m-2 h-1 bar-1, increasing by 45%; meanwhile, the Na2SO4/NaCl selectivity increased from 2.54 to 4.03. In addition, the fouling experiments illustrated that the fouling resistance of the zwitterion-modified NFMs to bovine serum albumin was significantly improved.

Ascending aortic dilation in adult patients with congenital ventricular septal defect: An observational study.

Many adult patients with congenital ventricular septal defect (VSD) also developed ascending aortic dilation, but few report the clinical features and surgical management of these patients. This study was designed to study ascending aortic dilation in adult patients with congenital VSD, and summarized the treatment experience and prognosis.To assess the clinical features and surgical management, we performed a retrospective analysis on preoperative data, intraoperative data, and postoperative data from the adult patients with congenital VSD who developed ascending aortic dilation in our institution from February 2010 to December 2016.From February 2010 to December 2016, we operated on 13 adult patients (12 males, 92.31%) with VSD who developed ascending aortic dilation. Median age was 37 (interquartile range 14) years. All patients suffered from perimembranous VSD and received surgical treatment. Their symptoms were all improved after surgery, no deaths occurred.Surgery is feasible for the ascending aortic dilation in adult patients with congenital VSD. Both proper perioperative treatment and close monitoring are required for the successful surgery.

CD40L promotes development of acute aortic dissection via induction of inflammation and impairment of endothelial cell function.

Acute aortic dissection is one of the most lethal cardiovascular disease. The major histopathological feature of AAD is medial degradation, especially breakdown of elastin and collagen. However, the underlying mechanism remains a mystery. Platelets expressed CD40 Ligand (CD40L) is recently recognised as a key effector of cardiovascular disease development through its pro-inflammatory effect. To clarify the role of CD40L in AAD, we examined level of CD40L in human blood serum samples and found that it is significantly higher in AAD patients compared with healthy subjects (26.8±5.52 ng/mL versus 13.4±4.00 ng/mL). To further investigate if CD40L is involve in the development of AAD, we applied ß-aminopropionitrile (BAPN) induced mouse model of AAD. Consistent with the human data, circulating CD40L in AAD mice much higher than normal mice (148.40±75.96 pg/mL versus 44.09±19.65 pg/mL). Meanwhile, multiple pro-inflammatory chemokines significantly increased in AAD mice. Importantly, the CD40L-/- mice treated with BAPN did not develop these phenotypes. Lastly, we confirmed that endothelial cells migration was significantly inhibited by CD40L, suggesting impaired recovery from intimal injury. In summary, we found that CD40L promoted AAD development through its pro-inflammatory effects and inhibition of endothelial cell function.

Subclinical hypothyroidism might worsen the effects of aging on serum lipid profiles: a population-based case-control study.

BACKGROUND: Dyslipidemia is an important global health problem, particularly in the elderly population. Traditionally, the high prevalence of dyslipidemia in elderly people is considered a "natural condition." Notably, subclinical hypothyroidism (SCH) is one of the most important risk factors for dyslipidemia. Few studies have assessed whether SCH plays a role in the increase in age-related dyslipidemia. This study aimed to explore the association between SCH and lipid profiles in different age groups. METHODS: This was a large-scale, population-based, case-control study. The population was derived from the REACTION study conducted across China. A total of 17,046 individuals (8827 cases and 8219 controls) aged 40 years or older were enrolled in the final analyses. The relationships between SCH and serum lipid parameters in each age group were evaluated after adjustment for thyroid hormones and common confounding factors. RESULTS: In the entire population, thyrotropin (TSH), the key indicator of SCH, was positively associated with cholesterol parameters (total cholesterol [TC] and low-density lipoprotein cholesterol [LDL-C]) through the sixth decade of life. After adjusting for common confounding factors and thyroid hormones, each 1 mIU/L increase in TSH was estimated to elevate the TC level by 0.0147 mmol/L and 0.0551 mmol/L, respectively, in individuals aged 40-49 years and 60-69 years. Similarly, with each 1 mIU/L increase in TSH, the LDL-C level tended to show gradually greater increases as age increased. In moderately old subjects (60-69 years), mild (TSH≤10 mIU/L) and significant (TSH>10 mIU/L) SCH increased the concentration of TC approximately 1.03- and 1.36-fold, and the concentration of LDL-C approximately 1.19- and 1.65-fold, respectively, when compared with younger subjects. CONCLUSIONS: TSH exhibited a stronger effect on the TC and LDL-C level in moderately old subjects than in younger subjects. SCH might augment and worsen the effects of aging on serum lipid profiles.

Thyroid-stimulating hormone levels within the reference range are associated with serum lipid profiles independent of thyroid hormones.

CONTEXT AND OBJECTIVE: Dyslipidemia in thyroid dysfunction has always been attributed to changes in thyroid hormone (TH) levels. We hypothesized that TSH plays an important role in lipid metabolism independent of TH. DESIGN AND SETTING: We conducted a cross-sectional study to investigate the relationship between serum TSH levels and lipid profiles after controlling for free T(3), free T(4), total T(3), total T(4) and nonthyroid factors relevant to lipid metabolism in euthyroid Chinese subjects. MAIN OUTCOME MEASURES: General linear analysis was performed to determine whether the impact of TSH on serum lipid levels is independent of the TH levels. Moreover, path analysis, an evolutionary multivariable regression technique, was conducted to test whether there is a direct and/or indirect effect between serum TSH and total cholesterol (TC) levels. Additionally, the odds ratios (95% confidence interval) for hypercholesterolemia in relation to TSH categories were calculated. RESULTS: A total of 3664 euthyroid subjects were finally analyzed. There was a significant linear trend toward higher log TC (P = 0.021) and log triglyceride (P = 0.001) levels with increasing serum TSH levels within the reference range, which remained significant after adjusting for factors such as TH levels, age, and smoking. Most importantly, the total effect of TSH on TC levels (total effect(TC, TSH) = 0.05253) includes a direct effect (direct effect(TC, TSH) = 0.05979) and an indirect effect via TH. Compared with subjects in the lower part of the reference range (TSH level, 0.27-0.61 mIU/liter), the adjusted odds ratio for hypercholesterolemia was 3.239 (95% confidence interval, 1.392-7.538; P = 0.007) for those in the upper category (TSH level, 4.61-5.5 mIU/liter). CONCLUSIONS: The variation in normal TSH levels is partially related to the lipid components and hypercholesterolemia in euthyroid subjects and includes both TH-dependent and TH-independent effects. Our study suggests the importance of controlling TSH in hypothyroid subjects.