[Expression and significance of INSM1 and SOX11 in pancreatic neuroendocrine tumor and solid pseudopapillary neoplasm].

OBJECTIVE: To investigate the expression and significance of insulinoma associated protein 1 (INSM1) and SRY-related high-mobility group box 11 (SOX11) in pancreatic neuroendocrine tumor (PNET) and solid pseudopapillary neoplasm (SPN). METHODS: To detect the expression of INSM1, SOX11, Syn, CgA, CD56, ß-catenin, and CD99 in 56 cases of PNET, 42 cases of SPN, 16 cases of ductal adenocarcinoma (DACC) and 8 cases of acinar cell carcinoma (ACC) by immunohistochemistry. The application value of combination of INSM1 and SOX11 was compared with conventional markers (Syn, CgA, CD56, ß-catenin, and CD99) in diagnosis and differential diagnosis of PNET and SPN. RESULTS: (1) In the 56 cases of PNET, the positive signals of INSM1 were located in the tumor and islet nucleus, the positive expression rate in the tumor tissues was 91.07% (51/56), whereas the signal was absent in 42 cases of SPN, 16 cases of DACC and 8 cases of ACC, and there were significant statistical difference between PNET with SPN, DACC, and ACC respectively (P < 0.001). (2) The positive signals of SOX11 were located in the tumor nucleus, with the positive expression rate was 92.86% (39/42) in SPN, however, the positive expression rate of SOX11 was 8.93% (5/56) in PNET, which included 3 cases of G1 and 2 cases of G3 types of PNET, the SOX11 positive signal was absent in 16 cases of DACC, 8 cases of ACC and peritumoral nomal pancreatic tissue, and the differences were statistically significant of positive rate between SPN with PNET, DACC and ACC, respectively (P < 0.001). (3) The sensitivity of INSM1(+)/SOX11(-) immunophenotype for PNET was 85.71%, vs. CD56 (57.14%), the difference was statistically significant (P=0.001); vs. Syn (80.36%) and CgA (71.43%), the difference was no statistically significant (P>0.05). The specificity of INSM1(+)/SOX11(-) for PNET was 100.00%, vs. Syn (42.86%) and CD56 (47.62%), the difference was statistically significant (P < 0.001); vs. CgA (92.86%), the difference was no statistically significant (P>0.05). The sensitivity of INSM1(-)/SOX11(+) immunophenotype for SPN was 92.86%, vs. ß-catenin (90.48%) and CD99 (85.71%), the difference was no statistically significant (P>0.05). The specificity of INSM1(-)/SOX11(+) for SPN was 96.43%, vs. CD99 (48.21%), the difference was statistically significant (P < 0.001); vs. ß-catenin (100.00%), the difference was no statistically significant (P>0.05). (4) The positive expression of INSM1 and SOX11 in PNET and SOX11 were not correlated with clinicopathological parameters (age, gender, tumor size, location, grade, and metastasis) (P>0.05). CONCLUSION: The positive expression patterns of INSM1 and SOX11 in PNET and SPN respectively are conductive to distinguish the both tumors. The combination of both take precedence over some corresponding conventional immunohistochemical markers in terms of sensitivity and specificity.

[Homeostatic medicine: new strategy and concept of health maintenance as well as diagnosis and treatment of diseases].

Homeostasis is a dynamic balance process of self-regulating. Biological systems remain stable through adapting to changing external conditions to maintain normal life activities. Homeostatic medicine is the science of studying homeostasis of human molecules, cells, organs and the whole body. It is a comprehensive discipline based on maintaining homeostasis to keep human health and assist for diseases prevention and diagnoses. Homeostatic medicine focuses on the whole body and on the role of homeostasis in health and disease, which is expected to provide new ideas and strategies for maintaining health as well as diagnosing and treating diseases. Nitric oxide (NO) plays an important role in the control of multisystem homeostasis. Nitrate is an important substance in regulating NO homeostasis through the nitrate-nitrite-NO pathway. Sialin, nitrate transporter which is located in the cell membrane and cytoplasm, mediates multiple cellular biological functions. The nitrate-nitrite-NO pathway and sialin-mediated biological functions play an important role in the regulation of body homeostasis.

[Expression of CD200 and INSM1 in gastrointestinal and pancreatic neuroendocrine neoplasms and its diagnostic values].

Objective: To investigate the expression and diagnostic values of CD200 and insulinoma associated protein 1 (INSM1) in gastrointestinal and pancreatic neuroendocrine neoplasm (GIP-NEN). Methods: The expression of CD200, INSM1, Syn and CgA was detected in 69 cases of GIP-NEN, 66 cases of gastrointestinal and pancreatic non-neuroendocrine neoplasm (GIP-nonNEN) and 16 cases of metastatic neuroendocrine neoplasm by immunohistochemistry, to compare the values of CD200, INSM1, Syn, CgA and their combinations in diagnosing GIP-NEN. Receiver operating characteristics (ROC) curve was used. Results: The immunoreactivity of CD200 was present in the cytoplasma and/or membrane of the neoplasms cells, the positive expression rates in GIP-NEN and GIP-nonNEN were significantly different (P<0.01). The sensitivity and specificity of CD200 for diagnosing GIP-NEN were 95.7% and 78.8%, respectively. There was significant difference of the positive rates of CD200 between neuroendocrine tumor and neuroendocrine carcinoma (P=0.05). The immunoreactivity of INSM1 was present in the nuclei of neoplasms cells. The positive expression rates in GIP-NEN and GIP-nonNEN were significantly different (P<0.01). The sensitivity and specificity of INSM1 for diagnosis of GIP-NEN were 85.5% and 95.5%, respectively. There were also significantly different positive rates of INSM1 between neuroendocrine tumor and neuroendocrine carcinoma, as well as between G1 and G3 neuroendocrine tumors (P<0.05). There was no difference in the area under ROC curve (AUC) of single stain of CD200, INSM1, Syn or CgA (0.857, 0.907, 0.890 and 0.833, respectively, P>0.05). The sensitivity of combined CD200+INSM1 stains for diagnosing GIP-NEN was significantly higher than that of Syn+CgA (85.5% vs. 63.8%, P<0.05). The AUC of two combinations were 0.962 and 0.925, respectively, which were not statistically different (P>0.05). Conclusions: CD200 and INSM1 are two novel markers of neuroendocrine neoplasm, which aid to diagnosis for GIP-NEN and exclude its mimickers. They are associated with tumor grades. Combining both as an immunohistochemical panel shows high sensitivity and specificity. Thus, the combined panel can be utilized as useful supplement for Syn and CgA.

[Clinical features and prognosis of patients with leptomeningeal metastases].

Objective: To describe the clinical manifestations, neuroimaging, cerebrospinal fluid(CSF) cytology and prognosis of Leptomeningeal metastases(LM). Methods: The clinical manifestations, imaging features and CSF cytology of LM patients admitted to Henan Provincial People's Hospital from May 1, 2015 to May 31, 2020 were retrospectively analyzed. The overall survival (OS) was evaluated by the time from the diagnosis of LM to death. Results: A total of 88 patients with LM were enrolled in the study, and the median age was 59 years (range:28-78 years). There were 42 males (47.7%) and 46 females (52.3%). According to the pathological classification, it was lung cancer in 58 cases (65.9%), gastric cancer in 13 cases (14.8%), breast cancer in 7 cases (8.0%), melanoma in 1 case, esophageal cancer in 1 case, gallbladder cancer in 1 case, renal cell carcinoma in 1 case, double source cancer in 2 cases, and unknown source in 4 cases. The median Karnofsky Performance Scale (KPS) score was 50. LM was the initial manifestation of cancer in 34 patients. All patients had LM-related clinical symptoms, including headache in 73 cases (83.0%), nausea and vomiting in 63 cases (71.6%), abnormal physical and mental behaviors in 37 cases (42.0%), seizure in 41 cases (46.6%). Cranial nerve involvement was observed in 23 patients (39.0%) and spinal nerve involvement in 20(33.9%). There were 61 patients (83.6%) who showed neuroimaging features of LM. Tumor cells or atypical cells were found in 90.8% of patients for the first time, and activated monocytes in 47 cases (54.7%). The median OS was 13.0 weeks (95%CI:2.9-23.1) with the 1-year survival rate of 19.1%. Univariate analysis of survival indicated that lung cancer, lower KPS score, tyrosine kinase inhibitors (TKIs) and whole brain radiotherapy were favorable predictors of survival (P<0.05). Conclusions: The overall prognosis of LM is poor. Good physical condition, TKIs treatment and whole brain radiotherapy might improve clinical outcomes of LM patients.

Investigation of the treatment modality in primary lymphoma of the salivary glands.

PURPOSE: To review our experience in managing primary lymphoma of the major salivary glands by analysing the treatment modality of patients. METHODS: In this study, we reviewed the medical records of 26 patients with primary malignant lymphoma of the major salivary glands who were treated at Beijing Stomatological Hospital of Capital Medical University from January 1999 to March 2015. The predictor variable was treatment modality. The outcome variables were overall survival (OS), disease-free survival (DFS), disease-special survival (DSS) and local control (LC). Other relevant variables were as follows: age, sex, pathological pattern, tumour site, clinical features, source, IPI and Ann-Arbor stage. The data were analysed by the χ2 test, log-rank test, and univariate analysis. RESULTS: Of the 26 patients, 14 received surgery only, four received initial surgery combined with adjuvant radiotherapy, five received adjuvant chemotherapy, and three received adjuvant radiochemotherapy. During the median follow-up period of 104 months, the overall estimates of OS, DFS, DSS and LC were 84.6, 84.6, 88.5 and 92.3%, respectively. Good outcomes were achieved in patients who received surgery combined with postoperative RT, followed by surgery only. MALT lymphoma had a better prognosis than the other pathological patterns. Mass, swelling, pathological pattern and Ann-Arbor stage were important prognostic factors. CONCLUSION: Surgery combined with postoperative radiotherapy may be an appropriate choice for patients with MALT lymphoma. The pathological pattern, mass, swelling and Ann-Arbor stage were associated with an unfavourable prognosis.

[Nitrate: a pioneer from the mouth to the systemic health and diseases].

Dietary nitrate which mainly comes from green leafy vegetables, is absorbed into blood circulation by the intestinal mucosa. Parotid gland is an important organ for transporting nitrate. Nitrate in blood is taken up by sialin, a nitrate transporter and concentrated in salivary glands and secreted into saliva. The salivary nitrate is partially reduced to nitrite and nitric oxide by oral bacteria, and then salivary nitrate and nitrite return into blood circulation with swallowing and intestinal mucosal absorption. As a non-classic source of nitric oxide, nitrate-nitrite-nitric oxide pathway plays an important role on physiological and pathological conditions, especially on the condition of hypoxia and ischemia. These functions include body protection, such as gastrointestinal tract, cardiovascular system, anti-inflammation, regulation of glucose/lipid metabolism, improvement of sport ability, maintaining gut microbiome hemostasis, and alleviating senility. The traditional view on nitrate as a harmful substance to human body has been proved to be lack of scientific evidence. With further research and application, as a pioneer from the mouth to the whole body, nitrate is expected to play a crucial part in human health, and prevention and treatment of systemic diseases.

[Pedigree analysis of C19ORF12 p.Asp18Tyr mutation in a family with mitochondrial membrane protein associated neurodegeneration].

Objective: To explore the clinical features and analyze the chromosome 19 open reading frame 12(C19ORF12) gene mutation of a family with mitochondrial membrane protein-associated neurodegeneration (MPAN). Methods: The pedigree diagnosed as neurodegeneration with brain iron accumulation (NBIA) in Henan Provincial People's Hospital in May 2018 was collected, and clinical data of the patients in this family was further analyzed. Furthermore, whole exome sequencing (WES) was employed to identify the disease-causing genes. Subsequently, the pathogenic mutation was validated by Sanger sequencing. Results: We identified 3 brothers, born of consanguineous Chinese parents. These patients were thought to carry autosomal recessive genes. The age of the onset ranged from 8 to 10 years old. All of the patients exhibited a chronic course, then got worse progressively. Parkinsonism was the first symptom. Other clinical features included cognitive decline, ataxia, gait abnormality, dysarthria and spastic paraplegia. All cases showed hypo-intensity in bilateral substantia nigra and globus pallidus on T(2)WI, FLAIR and SWI of MRI. However, the "eye-of-the-tiger sign" , which was commonly found in pantothenate kinase-associated neurodegeneration (PKAN), was absent. Further findings included cerebellar atrophy (all 3 patients) and the atrophy of temporal lobe (only one brother of the proband). The homozygous mutation c.52G>T (p.Asp18Tyr) was found through WES and Sanger sequencing. The proband's mother was heterozygous. This novel mutation was not reported in mutation database. Consequently, the pathogenic mutation in C19ORF12 gene (exon2, c.2327C>T, p.P776L) was identified from the patients according to the American College of Medical Genetics and Genomics (ACMG) guideline. The final diagnosis of the family was MPAN. Conclusions: We successfully identify a novel p.Asp18Tyr mutation in a family with MPAN. Our finding enriches the known MPAN mutation types and provides evidence for further research.

[Presenilin 1 p. F105L mutation was associated with a Chinese pedigree with early-onset familial Alzheimer's disease].

Objective: To investigate the clinical manifestations of an early-onset Alzheimer's family and the novel mutation sites of PSEN1 gene. Methods: Clinical data was collected from 30 members of a family with early onset Alzheimer's disease from Neurology Department of Henan Province People's Hospital in 2016, and high-throughput sequencing and sanger sequencing were used to detect and further validate the PSEN1 gene mutation. At the same time, 100 unrelated healthy people were selected as controls. Results: A total of 3 members of the family carried PSEN1 (p.F105L mutation). The clinical symptoms or imaging of the three people were consistent with the diagnosis of Alzheimer's disease. Conclusion: The p. F105L mutation of the PSEN1 gene has been found in the Chinese population, which may be involved in the pathogenesis of this familial Alzheimer's disease.

Prognostic value of pathologic grade for patients with oral squamous cell carcinoma.

OBJECTIVE: The purposes of this study were to explore both the prognostic value of pathologic grade and the relationships between differentiation and clinicopathological characteristics in oral squamous cell carcinoma. METHODS: This retrospective cohort study included the records of 2036 patients with oral squamous cell carcinoma who were surgically treated from June 1999 to December 2011. Chi-square test, Kaplan-Meier analysis, and Cox proportional hazards regression model were performed for statistical analysis. RESULTS: Many clinicopathological characteristics were associated with pathologic grade. Kaplan-Meier analysis showed that well-differentiated tumors had a better prognosis than the other two grades. Cox regression model showed that differentiation was an independent risk factor for prognosis in patients with early stage, but not with advanced stage. The predictive abilities of pathologic grade, T stage, N status, and lymph node ratio were similar, but the presence of extracapsular spread and perineural invasion were stronger prognostic factors than pathologic differentiation. CONCLUSIONS: Pathologic grade was found to be an independent risk factor for early-stage oral squamous cell carcinoma, but not for advanced stage. Many important clinicopathological characteristics were associated with histological classification; however, its prognostic value was limited.

From Nitrate to Nitric Oxide: The Role of Salivary Glands and Oral Bacteria.

The salivary glands and oral bacteria play an essential role in the conversion process from nitrate (NO3-) and nitrite (NO2-) to nitric oxide (NO) in the human body. NO is, at present, recognized as a multifarious messenger molecule with important vascular and metabolic functions. Besides the endogenous L-arginine pathway, which is catalyzed by complex NO synthases, nitrate in food contributes to the main extrinsic generation of NO through a series of sequential steps (NO3--NO2--NO pathway). Up to 25% of nitrate in circulation is actively taken up by the salivary glands, and as a result, its concentration in saliva can increase 10- to 20-fold. However, the mechanism has not been clearly illustrated until recently, when sialin was identified as an electrogenic 2NO3-/H+ transporter in the plasma membrane of salivary acinar cells. Subsequently, the oral bacterial species located at the posterior part of the tongue reduce nitrate to nitrite, as catalyzed by nitrate reductase enzymes. These bacteria use nitrate and nitrite as final electron acceptors in their respiration and meanwhile help the host to convert nitrate to NO as the first step. This review describes the role of salivary glands and oral bacteria in the metabolism of nitrate and in the maintenance of NO homeostasis. The potential therapeutic applications of oral inorganic nitrate and nitrite are also discussed.

PTX3 expression in the plasma of elderly ACI patients and its relationship with severity and prognosis of the disease.

OBJECTIVE: This study aims to investigate the expression of pentraxin3 (PTX3) in elderly patients with acute cerebral infarction (ACI) and to analyze the relationship of PTX3 with the severity and prognosis of ACI. PATIENTS AND METHODS: Between June 2014 and August 2015, 96 elderly patients with first-onset of ACI admitted to our institution were enrolled in the present study. Also, 70 healthy elderly subjects were included as controls in this study. Levels of PTX3, C-reactive protein (CRP), tumor necrosis factor (TNF)-α, homocysteine (Hcy), fibrinogen (FIB), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were determined in all patients both pre-therapeutically and 30 days post-therapeutically. Moreover, the severity of ACI was evaluated using the National Institute of Health stroke scale (NIHSS), and the prognosis was evaluated using the Modified Rankin Scale (mRS). The differences in the levels of above parameters were compared between groups, and the relationship between PTX3 and above biochemical parameters as well as the scores were investigated. RESULTS: PTX3 levels in the plasma of ACI patients were significantly higher than those of healthy controls (p < 0.05). Compared to low score group, mRS and levels of all parameters except Hcy and HDL-C were significantly increased in the patients of the high score group. In addition, plasma levels of HDL-C in the patients of the high score group were significantly lower than those in the low score group (p < 0.05). Biochemical parameters, NIHSS scores and mRS scores were significantly higher in the patients of the high concentration group than the low concentration group (p < 0.05), while no significant differences were observed in the plasma HDL-C levels between these two groups (p > 0.05). NIHSS scores and levels of all the biochemical parameters except HDL-C were significantly lower in the patients of the good prognosis group than in the patients of the poor prognosis group. HDL-C levels were significantly higher in the good prognosis group than in the poor prognosis group (p < 0.05). PTX3 levels were positively correlated with levels of CRP, TNF-α, Hcy, TC, TG and LDL-C as well as NIHSS score and mRS score, respectively, (r = 0.814, 0.682, 0.704, 0.726, 0.699, 0.734, 0.746, 0.753, p = 0.008, 0.043, 0.034, 0.027, 0.036, 0.024, 0.021, 0.019). However, no significant correlations were observed between FIB levels and HDL-C levels (r = 0.326, 0.626, p = 0.392, 0.071). CONCLUSIONS: Plasma levels of PTX3 are significantly elevated in elderly ACI patients, and the levels increase along with the exacerbation of the disease and deterioration in the prognosis. Also, PTX3 levels are positively correlated with levels of inflammatory markers as well as lipids. Therefore, PTX3 can be used as a biomarker for predicting and evaluating clinical conditions of ACI in elderly patients.

Transcriptome expression profiling of fur color formation in domestic rabbits using Solexa sequencing.

Fur color is an important, genetically determined characteristic of domestic rabbits, and rabbit furs are of great economic value. To investigate the molecular genetics associated with fur color determination in domestic rabbits, we used Solexa-sequencing technology to probe gene expression in dorsal skin tissues sampled from full-sibling Rex rabbits of different colors. The number of expressed genes in each sample was approximately 14,700. Among the top 30 genes and transcription factors with the highest reads per kilobase per million values, the elongation factor-alpha 1 gene was highly expressed in all samples, as were genes of the ribosomal protein and keratin gene families. Compared with the chinchilla (C) Rex rabbit control sample, the numbers of genes in the black (B) and white (W) rabbit samples were 1809 and 460, respectively, and the number of common differentially expressed genes was 257. Clustering analysis of these 257 genes revealed that 32 were up-regulated in sample B and down-regulated in sample W. Of these 32 genes, we identified some that are related to fur formation, including Tyrosinase-related protein 1 (TYRP1) and Tyrosinase (TYR), as well as genes with unknown functions. Quantitative real-time polymerase chain reaction was used to verify the expression patterns of those genes. The findings are expected to provide reference for the further study of fur color formation in rabbits.