CD11c+ B Cells Participate in the Pathogenesis of Graves' Disease by Secreting Thyroid Autoantibodies and Cytokines.

Graves' disease (GD) is a common autoimmune disorder with an elevation in pathogenic autoantibodies, specifically anti-thyrotropin receptor antibodies (TRAbs), which are secreted by autoreactive B cells. To date, there has been little research on self-reactive B cells in GD. In the current study, we reported that a unique B-cell subset, CD11c+ B cells, was expanded in the peripheral blood (PB) of GD patients, as detected by flow cytometry. The frequency of CD11c+ B cells was positively correlated with serum TRAb levels. The flow cytometry data showed that CD11c expression was higher in a variety of B-cell subsets and that CD11c+ B cells presented a distinct immunophenotype compared to paired CD11c- B cells. Immunohistochemical and immunofluorescence staining indicated the presence of CD11c+CD19+ B cells in lymphocyte infiltration areas of the GD thyroid. Flow cytometric analysis of PB and fine-needle aspiration (FNA) samples showed that compared to PB CD11c+ B cells, CD11c+ B cells in the thyroid accumulated and further differentiated. We found that CD11c+ B cells from the PB of GD patients were induced to differentiate into autoreactive antibody-secreting cells (ASCs) capable of secreting TRAbs in vitro. Luminex liquid suspension chip detection data showed that CD11c+ B cells also secreted a variety of cytokines, including proinflammatory cytokines, anti-inflammatory cytokines, and chemokines, which might play roles in regulating the local inflammatory response and infiltration of lymphocytes in the thyroid. In addition, we performed a chemotaxis assay in a Transwell chamber to verify that CD11c+ B cells were recruited by thyroid follicular cells (TFCs) via the CXCR3-CXCL10 axis. In conclusion, our study determined that CD11c+ B cells were involved in the pathogenesis of GD in multiple ways and might represent a promising immunotherapeutic target in the future.

Hydrogen Sulfide Promotes Thyroid Hormone Synthesis and Secretion by Upregulating Sirtuin-1.

Objective: One mechanism of hypothyroidism involves the disruption of thyroid hormone synthesis and secretion by thyrocytes. Hydrogen sulfide (H2S), as a gas signaling molecule, participates in many physiopathologic processes by upregulating sirtuin-1 (SIRT1). The aim of the current study was to explore whether H2S promotes the synthesis and secretion of thyroid hormones by upregulating SIRT1. Methods: Real-time PCR and immunohistochemistry were used to detect the mRNA and protein expression of H2S-generating enzymes in normal human thyroid tissues. Serum H2S concentrations from hypothyroid patients (n = 32) and euthyroid participants (n = 41) were detected by H2S-selective sensors. Thirty-one Sprague-Dawley rats were divided into control group (n = 10), hypothyroid group (induced by MMI, n = 10) and hypothyroid + NaHS group (n = 11), and the FT4, TT4 and TSH levels were assayed. Human primary thyrocytes were incubated with H2S donor sodium hydrosulfide (NaHS) or NaHS plus SIRT1 inhibitor (EX527) in vitro. Thyroid hormone synthesis- and secretion-related proteins [thyroid peroxidase (TPO), sodium iodide transporter (NIS), Pendrin, monocarboxylic acid transporter 8 (MCT8)] were analyzed by real-time PCR and Western blot. Results: H2S levels in serum from hypothyroid patients were decreased compared to those from euthyroid participants (p < .05), and serum H2S levels were positively correlated with FT3, FT4, TT3, and TT4 levels in all subjects (all p < .0001). In vivo, NaHS promoted thyroid function in hypothyroid rats (p < .05). In vitro, H2S was detected in supernatant, and CBS mRNA was higher than CSE and 3-MPST in human primary thyrocytes (p < .05). The protein levels of TPO, NIS, Pendrin and MCT8 were upregulated in a concentration-dependent manner for NaHS in thyrocytes. After blocking SIRT1 with EX527, we found that the increasing levels of TPO, NIS, Pendrin, and MCT8 and TPO activity were downregulated in thyrocytes incubated with NaHS, and FT4 levels in the cell supernatant were also decreased significantly (all p < .05). Conclusion: H2S is mainly generated in thyrocytes by CBS. Serum H2S levels are decreased with hypothyroidism. H2S promotes the synthesis and secretion of thyroid hormones and the expression of related molecules by upregulating SIRT1.

Validation of the CPS+EG and Neo-Bioscore staging systems after preoperative systemic therapy for breast cancer in a single center in China.

BACKGROUND: Prognostic assessment after preoperative systemic therapy (PST) plays a vital role in determining treatment in breast cancer patients. Many researchers have sought to develop a system to quantitate residual tumor and its correlation with prognosis after PST. This retrospective study validated the CPS + EG staging system and Neo-Bioscore in a single center in China. METHODS: Data from patients with non-metastatic primary breast cancer who were treated with PST and surgery from Jan. 2008 to Dec. 2014 at the Breast Disease Center of Peking University First Hospital, China, were reviewed. DFS, DSS and OS were calculated using the K-M curve and AUC. Multivariate analysis was used for a Cox proportional hazards model. All calculations were performed with SAS 9.4. RESULTS: A total of 403 patients were enrolled in this study. The median follow-up period was 45 (range 11-107) months. The five-year DFS, DSS and OS rates were 86.4%, 91.2% and 90.5%, respectively. The CS, PS, CPS + EG staging system and Neo-Bioscore stratified patients according to DFS, DSS, and OS after PST, with all P values < 0.0001. The CPS + EG staging system and Neo-Bioscore stratified prognosis after PST better than CS. HER2-positive patients without trastuzumab treatment had obviously worse DFS and OS than other subgroups with different HER2 statuses that scored a 3 in the Neo-Bioscore system. CONCLUSIONS: The CPS + EG staging system and Neo-Bioscore can improve prognostic prediction in non-pCR breast cancer patients after PST and, provided unfavorable prognostic factors such as insufficient treatment are incorporated, will have broader clinical applicability.

Effects of tissue factor, PAR-2 and MMP-9 expression on human breast cancer cell line MCF-7 invasion.

OBJECTIVE: This study aimed to explore the expression of tissue factor (TF), protease activated receptor-2 (PAR-2), and matrix metalloproteinase-9 (MMP-9) in the MCF-7 breast cancer cell line and influence on invasiveness. METHODS: Stable MCF-7 cells transfected with TF cDNA and with TF ShRNA were established. TF, PAR-2, and MMP-9 protein expression was analyzed using indirect immunofluorescence and invasiveness was evaluated using a cell invasion test. Effects of an exogenous PAR-2 agonist were also examined. RESULTS: TF protein expression significantly differed between the TF cDNA and TF ShRNA groups. MMP-9 protein expression was significantly correlated with TF protein expression, but PAR-2 protein expression was unaffected. The PAR- 2 agonist significantly enhanced MMP-9 expression and slightly increased TF and PAR-2 expression in the TF ShRNA group, but did not significantly affect protein expression in MCF-7 cells transfected with TF cDNA. TF and MMP-9 expression was positively correlated with the invasiveness of tumor cells. CONCLUSION: TF, PAR-2, and MMP-9 affect invasiveness of MCF-7 cells. TF may increase MMP-9 expression by activating PAR-2.

[Relationship between Ki67 expression and tumor response to neoadjuvant chemotherapy with anthracyclines plus taxanes in breast cancer].

OBJECTIVE: To investigate the relationship between Ki67 expression and tumor response to neoadjuvant chemotherapy with anthracyclines plus taxanes in breast cancer. METHODS: From January 2008 to June 2009, 129 patients with primary breast invasive ductal cancer received neoadjuvant chemotherapy with anthracyclines plus taxanes. The expression of Ki67 in the tumor tissues was determined by using immunohistochemistry with core needle biopsy specimens prior to the chemotherapy. The tumor response to the chemotherapy was evaluated by dynamic enhanced MRI based on RECIST2000 criteria, pathologic response was assessed according to Miller-Payne grading system, and the clinical comprehensive response was evaluated based on MRI combined with pathologic response. RESULTS: Dynamic enhanced MRI classified 87 cases (67.4%) as effective. According to the Miller-Payne grading system, 99 cases (76.7%) were ranged effective. One hundred and ten cases (85.5%) were recognized as clinically comprehensive effective. The effective rates of neoadjuvant chemotherapy in patients with a Ki67 expression >10% evaluated by the above-mentioned three standards were 73.2%, 81.4% and 89.7%, respectively; and those in patients with a Ki67 expression < or = 10% were 50.0%, 62.5% and 71.9%, respectively. Compared with patients with a Ki67 expression < or = 10%, the patients with a Ki67 expression >10% had better response rates determined by all the three standards (P values were 0.020, 0.030 and 0.010, respectively). The Ki67 expression in the tumor tissue was linearly correlated with clinically comprehensive response on the Linear-Linear association analysis. CONCLUSIONS: There is a statistic association between Ki67 expression and tumor response to the neoadjuvant chemotherapy with anthracyclines plus taxanes in breast cancer, and the patients with a higher expression of Ki67 has a better tumor response to the chemotherapy.

Analysis of risk factors of surgical site infections in breast cancer.

BACKGROUND: Adjuvant chemotherapy has become an important component of standard therapy for breast cancer. However, until now, there have been few reports on the surgical site infections (SSI) after breast cancer surgery, specially after adjuvant chemotherapy. To study the risk factors of SSI of breast cancer, we analyzed patients diagnosed with breast cancer and treated with surgery. METHODS: Fifty-five patients diagnosed with breast cancer and received breast conserving or modified radical operations in our hospital during January 2008 to March 2008 were selected. Factors (patients' age, body mass index (BMI), diabetes mellitus, no or administered adjuvant chemotherapy, with or without onset of myelosuppression and the degree, surgical approaches, duration of operation, postoperative drainage duration and total drainage volume) associated with SSI were retrospectively reviewed and statistically analyzed by single factor analysis. RESULTS: Five patients suffered SSI (5/55, 9.1%); nineteen receiving adjuvant chemotherapy experienced Grade III + myelosuppression, among which 4 had SSI; only 1 out of the remaining 36 patients without adjuvant chemotherapy had SSI. The difference between the two groups was significant (P = 0.043). The incidence of SSI in patients with post-operative drainage tube indwelling longer than 10 days was 5/21, whereas no SSI occurred in that less than 10 days (P = 0.009). In our study, there was no significant difference in other associated factors. CONCLUSIONS: Concurrent Grade III + myelosuppression after adjuvant chemotherapy is an important risk factor of SSI in breast cancer and needs further study. No SSI was detected with indwelling time of post operative drainage less than 10 days.

[Prospective study on the role of MRI and B ultrasonography in evaluating the tumor response to neoadjuvant chemotherapy in breast cancer].

OBJECTIVE: To evaluate the role of breast B ultrasonography and magnetic resonance imaging in assessing the tumor response to neoadjuvant chemotherapy in breast cancer. METHODS: Eighty-five patients with breast cancer diagnosed by core needle biopsy received neoadjuvant chemotherapy entered this prospective study. Breast B ultrasonography and dynamic enhanced MRI was performed before chemotherapy induction, after the second course and the fourth course of chemotherapy prior to the surgery. Clinical evaluation was made through the tumor reduction measured by B ultrasonography and MRI, based on the response evaluation criteria in solid tumors (RECIST). RESULTS: Measured by dynamic enhanced MRI, 56 patients got partial response (PR), 27 got stable disease (SD) and 2 got progressive disease (PD), none complete response (CR). Measured by B ultrasonography, 52 patients got PR, 31 got SD, 2 got PD, no CR. Residual tumor size after chemotherapy on MRI correlated well with post-operative pathologic findings (r = 0.783, P < 0.05), and B ultrasonography correlated moderately with microscopic findings (r = 0.576, P < 0.001). CONCLUSION: Dynamic enhanced MRI is a reliable method to evaluate tumor response to neoadjuvant chemotherapy in breast cancer.

Paraneoplastic pemphigus associated with Castleman tumor: a commonly reported subtype of paraneoplastic pemphigus in China.

BACKGROUND: Castleman tumor, a rare lymphoproliferative disorder, is one of the associated tumors in paraneoplastic pemphigus. We analyzed the characteristics of a group of patients with Castleman tumor to clearly understand and to improve the prognosis of the disease. OBSERVATIONS: Ten cases of paraneoplastic pemphigus associated with Castleman tumor treated in the Department of Dermatology, Peking University First Hospital, Beijing, China, from May 1, 1999, to March 31, 2004, were analyzed for clinical aspects, characteristics and histologic features of the tumors, and computed tomographic findings. Literature was reviewed and data were compared with our cases. Castleman tumor was a frequently reported neoplasm in association with paraneoplastic pemphigus in China. The disease was found to be caused by an autoimmune reaction originating from the B lymphocytes in the Castleman tumor. CONCLUSIONS: Castleman tumor in association with paraneoplastic pemphigus is a commonly reported subtype of paraneoplastic pemphigus in China. Early detection and removal of the Castleman tumor are crucial for the treatment of this tumor-associated autoimmune disease.

[Treatment analysis of 26 patients with breast ductal carcinoma in situ].

OBJECTIVE: To study the appropriate surgical treatment for breast ductal carcinoma in situ (DCIS). METHODS: Twenty-six such patients treated between 1992 and 2001 were retrospectively analyzed. Among them, 3 patients were treated by simple mastectomy, 23 patients by mastectomy and axillary lymph node dissection, 8 patients by chemotherapy and one patient by radiotherapy after operation. Median follow-up was 42 m (rang 12 - 112 m). RESULTS: Except 3 of these 26 patients lost in follow-up and 1 patient died from diabetes mellitus, all the other 22 patients survived over 5 years. All lymph nodes dissected from 23 patients were negative. After surgery, 3 patients developed lymph edema of the arm. CONCLUSION: DCIS, lacking the potential of metastasis, is not invasive. Conservative breast surgery without lymph node dissection is feasible for most DCIS patients.