Supplementary Material for: Response Predictors to Calcineurin Inhibitors in Patients with Primary Membranous Nephropathy

<b><i>Background:</i></b> Currently, there is an urgent need to find ways of identifying primary membranous nephropathy (PMN) patients who are likely to benefit from calcineurin inhibitors (CNI) or who are resistant to them. In this study, we employed nano-HPLC-MS/MS analysis to identify serum biomarkers that predict the clinical response to CNI therapy in PMN patients. <b><i>Methods:</i></b> The endpoint was complete remission (CR) after CNI treatment. PMN patients were grouped into no-remission (NR) or CR groups to screen predictive candidates using the nano-HPLC-MS/MS analysis. <b><i>Results:</i></b> Compared with NR patients, 3 upregulated proteins and 5 downregulated proteins were found to present a twofold change in CR patients’ serum. Serum amyloid A1 protein (SAA1) was further validated by ELISA; it was decreased in patients in the NR group compared with patients in the CR group, but SAA1 in patients in these groups was lower than in healthy controls and minimal change disease patients. The area under the receiver operating characteristic (ROC) curve of SAA1 was used to distinguish PMN NR patients from those in remission and was 0.901, with a sensitivity of 78.3% and specificity of 86.8%, similar to that of the phospholipase A2 receptor (PLA2R) antibody. Combining SAA1 with the PLA2R antibody, the area under the ROC curve was 0.956, which was higher than that of SAA1 or the PLA2R antibody alone. <b><i>Conclusions:</i></b> Serum SAA1 may be a candidate PMN biomarker that can be used to discriminate CNI NR cases from remission patients. The combination of SAA1 and the PLA2R antibody increases the accuracy of diagnosis

Supplementary Material for: Correlation of Neuroendocrine Differentiation with A Distinctively Suppressive Immune Microenvironment in Gastric Cancer

Introduction Neuroendocrine neoplasms (NENs) harbored significantly suppressive tumor immune microenvironments (TIMEs). However, the immunological effects of neuroendocrine differentiation (NED) on non-NE neoplasms, such as gastric cancer (GC), were unknown. Methods Between pure gastric cancer (PGC) and GC-NED, TIME features were scored based on expression data and validated on serial whole-tissue sections of surgical samples, with tertiary lymphoid structures (TLSs) and the extra-TLS zone evaluated independently using multi-marker immunohistochemical staining. Risk analyses of TIME features on tumor behaviors were performed in GC-NED. The universal immunological effects of NED were explored preliminarily in adenocarcinomas arising in other organs. Results Based on over 11,500 annotated TLSs and 2,700 extra-TLS zones, compared with PGC, GC-NED harbored a distinctively more suppressive TIME characterized by increased but immature TLSs, with higher naïve B cell and follicular regulatory T cell densities and downregulated TLS maturation-related cell ratios inside TLSs; increased naïve B cell and regulatory T cell densities and a high proportion of exhausted T cells in the extra-TLS zone. The upregulated tumor PD-L1 expression and its close correlations with TLS formation and maturation were remarkable exclusively in GC-NED. TIME features, especially those regarding TLSs, were significantly correlated with tumor growth and invasion. The desynchrony between TLS formation and maturation and increased naïve or regulatory immune cell infiltration was observed in adenocarcinomas of the colorectum, pancreas, lung, and prostate. Conclusion NED highlighted a distinct GC entity with more suppressive TIME features correlated with tumor behaviors, indicating a cohort that would benefit more from immunotherapies

Supplementary Material for: Correlation of Neuroendocrine Differentiation with A Distinctively Suppressive Immune Microenvironment in Gastric Cancer

Introduction Neuroendocrine neoplasms (NENs) harbored significantly suppressive tumor immune microenvironments (TIMEs). However, the immunological effects of neuroendocrine differentiation (NED) on non-NE neoplasms, such as gastric cancer (GC), were unknown. Methods Between pure gastric cancer (PGC) and GC-NED, TIME features were scored based on expression data and validated on serial whole-tissue sections of surgical samples, with tertiary lymphoid structures (TLSs) and the extra-TLS zone evaluated independently using multi-marker immunohistochemical staining. Risk analyses of TIME features on tumor behaviors were performed in GC-NED. The universal immunological effects of NED were explored preliminarily in adenocarcinomas arising in other organs. Results Based on over 11,500 annotated TLSs and 2,700 extra-TLS zones, compared with PGC, GC-NED harbored a distinctively more suppressive TIME characterized by increased but immature TLSs, with higher naïve B cell and follicular regulatory T cell densities and downregulated TLS maturation-related cell ratios inside TLSs; increased naïve B cell and regulatory T cell densities and a high proportion of exhausted T cells in the extra-TLS zone. The upregulated tumor PD-L1 expression and its close correlations with TLS formation and maturation were remarkable exclusively in GC-NED. TIME features, especially those regarding TLSs, were significantly correlated with tumor growth and invasion. The desynchrony between TLS formation and maturation and increased naïve or regulatory immune cell infiltration was observed in adenocarcinomas of the colorectum, pancreas, lung, and prostate. Conclusion NED highlighted a distinct GC entity with more suppressive TIME features correlated with tumor behaviors, indicating a cohort that would benefit more from immunotherapies

Supplementary Material for: Correlation of Neuroendocrine Differentiation with A Distinctively Suppressive Immune Microenvironment in Gastric Cancer

Introduction Neuroendocrine neoplasms (NENs) harbored significantly suppressive tumor immune microenvironments (TIMEs). However, the immunological effects of neuroendocrine differentiation (NED) on non-NE neoplasms, such as gastric cancer (GC), were unknown. Methods Between pure gastric cancer (PGC) and GC-NED, TIME features were scored based on expression data and validated on serial whole-tissue sections of surgical samples, with tertiary lymphoid structures (TLSs) and the extra-TLS zone evaluated independently using multi-marker immunohistochemical staining. Risk analyses of TIME features on tumor behaviors were performed in GC-NED. The universal immunological effects of NED were explored preliminarily in adenocarcinomas arising in other organs. Results Based on over 11,500 annotated TLSs and 2,700 extra-TLS zones, compared with PGC, GC-NED harbored a distinctively more suppressive TIME characterized by increased but immature TLSs, with higher naïve B cell and follicular regulatory T cell densities and downregulated TLS maturation-related cell ratios inside TLSs; increased naïve B cell and regulatory T cell densities and a high proportion of exhausted T cells in the extra-TLS zone. The upregulated tumor PD-L1 expression and its close correlations with TLS formation and maturation were remarkable exclusively in GC-NED. TIME features, especially those regarding TLSs, were significantly correlated with tumor growth and invasion. The desynchrony between TLS formation and maturation and increased naïve or regulatory immune cell infiltration was observed in adenocarcinomas of the colorectum, pancreas, lung, and prostate. Conclusion NED highlighted a distinct GC entity with more suppressive TIME features correlated with tumor behaviors, indicating a cohort that would benefit more from immunotherapies

Supplementary Material for: Contralateral Hippocampal Stimulation for Failed Unilateral Anterior Temporal Lobectomy in Patients with Bilateral Temporal Lobe Epilepsy

<i>Aims:</i> To prospectively study the surgical outcomes of unilateral anterior temporal lobectomy (ATL) in patients with intractable bilateral temporal lobe epilepsy (TLE) as well as two-staged contralateral hippocampal stimulation in patients after failed unilateral ATL. <i>Methods:</i> Eighteen carefully selected patients with bilateral TLE underwent unilateral ATL. Five cases with failed ATL underwent two-staged contralateral hippocampal stimulation. Seizure control and changes in intelligence quotient (IQ), memory quotient, and quality of life (QOL) were analyzed 2-5 years after treatment. <i>Results:</i> In the patients with unilateral ATL, the percentages seizure free were 55.6% (10/18), 50.0% (9/18), and 44.4% (4/9) at the 1-, 2-, and 5-year follow-up visits, respectively. There were significant difference in seizure control between the patients with unilateral ATL and the 12 cases in the medication group. Significant differences were also found in changes in the patients' QOL and full-scale IQ at the 2-year follow-up between the surgical and medication groups. Five patients who underwent contralateral hippocampal stimulation after failed unilateral ATL experienced 80-100% seizure reductions, and 80% were seizure free 1 year after hippocampal stimulation. <i>Conclusion:</i> Unilateral ATL provides good seizure control and does not cause serious memory or IQ injury in carefully selected patients with true bilateral TLE. Contralateral hippocampal stimulation is a useful approach for patients who experience unilateral ATL failure

Supplementary Material for: Role of Epidermal Growth Factor Receptor Expression on Patient Survival in Pancreatic Cancer: A Meta-Analysis

<p><i>Background/Aims:</i> Epidermal growth factor receptor (EGFR) has been considered as an attractive and potential therapeutic target of pancreatic cancer. However, the clinical importance of EGFR expression remains controversial. <i>Methods:</i> We performed a meta-analysis of previous studies to quantitatively review the effects of EGFR expression on survival in patients with pancreatic cancer. <i>Results:</i> Eight studies (570 patients) were included to perform a meta-analysis of the survival results. Overall, positivity for EGFR expression was 45.1% in pancreatic carcinoma. The combined hazard ratio was 1.225 (95% CI 1.014–1.481; p = 0.035), indicating that EGFR expression has a significant impact on survival. Heterogeneity was absent between studies and publication bias, which suggests that the summary statistics obtained may approximate the actual average. Three trials reported a survival disadvantage for patients with EGFR expression while five trials reported no statistically significant difference. <i>Conclusion:</i> EGFR expression is a poor prognostic factor for survival in patients with pancreatic cancer.</p

Supplementary Material for: Initiation and Cessation Timing of Renal Replacement Therapy in Patients with Type 1 Cardiorenal Syndrome: An Observational Study

<p><b><i>Background/Aims:</i></b> Renal replacement therapy (RRT) is a rescue therapy for patients with type 1 cardiorenal syndrome (CRS) with poor prognoses. However, the optimal timing for initiation and cessation of RRT remains controversial. The purpose of this study was to determine the optimal timing of initiation and cessation of RRT for patients with type 1 CRS. <b><i>Methods:</i></b> In this retrospective analysis, patients with refractory type 1 CRS receiving RRT were divided into 3 groups according to weaning from RRT and death within 90 days. Baseline characteristics, underlying heart disease, comorbidities, drug use before RRT, indicators of RRT initiation, and prognosis were compared between the 3 groups. <b><i>Results:</i></b> Fifty-two patients were enrolled, which included 27 males and 25 females with a mean age of 70.7 ± 16.1 years and a 90-day mortality rate of 65.4%. The mean urine output before RRT initiation was 800 mL/ 24 h in the RRT-independent group, 650 mL/24 h in the RRT-dependent group, and 345 mL/ 24 h in the death group (<i>p</i> = 0.021). Additionally, there were obvious differences in fluid balance between the 3 groups (167, 250, and 1,270 mL, respectively, <i>p</i> = 0.016). Patients could be successfully weaned from RRT when urine output was >880 mL and fluid balance volume was <150 mL. <b><i>Conclusion:</i></b> The mean fluid balance of survivors was remarkably less than that of the death group at RRT initiation. RRT termination can be considered when urine output is >880 mL/24 h and volume balance is <150 mL/24 h.</p

Supplementary Material for: Novel Uromodulin Mutation in Familial Juvenile Hyperuricemic Nephropathy

<b><i>Background:</i></b> Familial juvenile hyperuricemic nephropathy (FJHN) is an autosomal dominant disorder characterized by early onset of hyperuricemia, decreased fractional renal urate excretion and progressive interstitial nephropathy. Mutations in the uromodulin<i> (UMOD)</i> gene encoding uromodulin/Tamm-Horsfall, a glycosylphosphatidylinositol (GPI)-anchored protein, cause this disease. <b><i>Methods:</i></b> One Chinese family with 13 FJHN-affected individuals is described. Clinical data, blood and urine samples of 7 affected members (all alive patients in this family) and 15 unaffected members were collected. Mutation analysis of the <i>UMOD</i> gene was performed by polymerase chain reaction and direct sequencing. Urinary uromodulin from affected or unaffected members of this family and healthy controls was examined by enzyme-linked immunosorbent assay kit. Expression of uromodulin in renal tissue was shown with immunofluorescence. <b><i>Results:</i></b> A novel mutation (p.T605G) within the uromodulin GPI anchor signal segment was identified in the affected individuals of this FJHN family. There was a markedly increased expression of uromodulin in renal tissue and significantly decreased urinary excretion of uromodulin in affected patients with an estimated glomerular filtration rate <60 ml/min/1.73 m². <b><i>Conclusions:</i></b> The present study reported a novel mutation in exon 9 of <i>UMOD</i> in the Chinese Han population, within the GPI anchor signal segment of uromodulin. Since the GPI anchor is linked with the release or secretion of proteins, our finding may provide further evidence for the underlying mechanism of decreased urinary excretion of uromodulin in FJHN

Supplementary Material for: Relationship among Mortality of Patients with Acute Kidney Injury after Cardiac Surgery, Fluid Balance and Ultrafiltration of Renal Replacement Therapy: An Observational Study

<p><b><i>Background/Aims:</i></b> The study aimed to investigate the relationship among mortality of patients with cardiac surgery-associated acute kidney injury (CSA-AKI), fluid balance, and ultrafiltration of renal replacement therapy (RRT). <b><i>Methods:</i></b> From January 2009 to October 2015, hospitalized patients with CSA-AKI receiving continuous or prolonged intermittent RRT were screened. The effects of fluid balance and ultrafiltration of RRT on clinical outcome were analyzed. <b><i>Results:</i></b> The 30-day mortality of all the 63 patients in the study was 58.6%. Compared with the death group, the survival group had a significantly lower fluid balance, larger ultrafiltration volume, and similar ultrafiltration rate during the first 3 days of RRT. Multivariate Cox regression analysis revealed that positive fluid balance during the first day of RRT, cardiac function of grade IV, and higher Sequential Organ Failure Assessment score were independent risk factors of 30-day mortality. <b><i>Conclusion:</i></b> Fluid balance was more relevant to short-term prognosis of CSA-AKI-RRT patients than ultrafiltration volume or ultrafiltration rate.</p

Supplementary Material for:Effect of Cardiac Surgery-Associated Acute Kidney Injury on Long-Term Outcomes of Chinese Patients: A Historical Cohort Study

<i>Background/Aims:</i> To evaluate the long-term outcomes of Chinese patients with cardiac surgery-associated acute kidney injury (CSA-AKI). <i>Methods:</i> Patients who underwent cardiac surgery with a median 3-year follow-up were enrolled. The long-term survival rate and the incidence of chronic kidney disease (CKD) were recorded, and related risk factors were analyzed. <i>Results:</i> Of all 1,363 patients, 457 (33.5%) developed CSA-AKI. The AKI patients had a lower 3-year survival rate (88.8 vs. 97.2%, respectively, <i>p</i> < 0.001) and a higher incidence of CKD stages 3-5 (9.9 vs. 2.3%, respectively, <i>p</i> < 0.001) than the non-AKI patients. Cox regression analysis showed that AKI, atrial fibrillation, chronic cardiac insufficiency, longer surgical duration, respiratory failure after surgery, and longer mechanical ventilation time were associated with long-term mortality, while AKI, older age, and lower baseline kidney function were associated with incident CKD stages 3-5. <i>Conclusion:</i> CSA-AKI increased the risk of 3-year mortality and incident CKD stages 3-5