Aim To improve the sensitivity of phycobiliprotein immunofluorescence assay(PBPIFA), by introducing into bridged avidin biotin(BRAB) technique,known as BRAB-PBPIFA. Methods Using ELISA as a gold standard, effectiveness of three assays(direct PBPIFA, BRAB-PBPIFA and ELISA)were compared in simultaneous detection of standard antigen and 500 serum samples. Results BRAB improved greatly sensitivity of PBPIFA in detecting antigen of low concentration, and detection rate of weakly positive samples could increase by one times and a half. In the applying, protein-free blocking agent Tween 20 to BRAB-PBPIFA,possessed better blocking effect. Conclusion Combination of BRAB technique with PBPIFA can improve the detcctive sensitivity.

AIM:To observe the effects of supraoptic ADH neurons and central diabetes insipidus(CDI)in Wistar rats at different times after hypophysectomy.METHODS:Hypophysectomy was undergone by stereotaxic instrument.Water intake,urine output and urine specific gravity(SG)were observed each day.The survival rate of supraoptic ADH neurons was determined by immunofluorescence after hypophysectomy at different times.RESULTS:The rats manifested triphasic CDI after hypophysectomy.The average water intake in experiment group was 73.9 mL vs 30.9 mL in control group(P0.05),but the cellular body hypertrophy appeared.The survival rate at 10th day was 72%(P0.05),but they were all less than the 10th day(P

ObjectiveTo investigate the relationship between syndrome differentiation types and the objective indexes of fatty liver.MethodsA cross-sectional study was adopted. Clinical observation table of fatty liver for TCM diagnostic performance was used to collect the data of patients and cluster analysis was adopted for syndrome differentiation. The difference of the objective indexes among different syndromes was studied.ResultsFatty liver showed more severity in spleen Qi deficiency and dampness group compared with heart and liver yin deficiency group(χ2=8.218,P=0.041). Patients in the spleen and stomach Qi deficiency group had less smoking history(χ2=8.416,P=0.038). Patients in the spleen Qi deficiency and dampness group had higher AST, TP, ALP and WHR indexes and lower Alb.ConclusionsDifferences of objective indexes in different fatty liver syndromes can be used for enriching syndrome differentiation contents, and provide the basis for microcosmic syndrome differentiation of fatty liver.

Objective To investigate the function of the hypothalamic-pituitary-adrenal (HPA) axis in postmenopausal women with type 2 diabetes and visceral obesity. Methods Subjects were divided into three groups:control group(group C),type 2 diabetes mellitus with non-obesity group (group DM) and type 2 diabetes mellitus with visceral obesity group (group DM + OB). General clinical characteristics, morning blood cortisol concentrations and 24 h urine free cortisol of three groups were compared. Serum cortisol levels were also compared after 0.25 mg dexamethasone suppression test and followed by oral intake of 25 mg cortisone acetate. Results (1) There were no significant differences in basal cortisol levels, but after inhibition with dexamethasone the group DM + OB showed significantly higher cortisol level than that in the control group (P < 0.01). (2) Conversion of oral cortisone to plasma cortisol differed significantly between the group C (lower) and group DM + OB (P < 0.05). (3) Plasma LH and FSH concerntrations were significantly lower in group DM + OB compared with group C (P < 0.01). Conclusion In the postmenopausal women with type 2 diabetes mellitus, the negative feedback mechanism and hepatic 11β-HSD-1 activity were impaired, especially in those with visceral obesity.

The plasma level and the regulation of cortisol in type 2 diabetic patients were invesligated.Plasma and urinary cortisol levels were measured, and dexamethasonc suppression test and oral cortisone test in vivo were performed. Compared with controls, diabetic patients had higher urinary cortisol level. The activity of hepatic 11 β-hydroxy-steroid dehydrogenase-1 (11β-HSD1) in type 2 diabetic patients was decreased, suggesting that the elevated basal cortisol in type 2 diabetic patiens may due to impaired hepatic degradation of cortisol.

Objective:To study the changes of food characters in early swallowing recovery in patients with different laryngeal function preservation surgery.Methods:Collected patients with laryngeal cancer hypopharyngeal cancer who underwent laryngeal function preservation surgery in Lihuili Hospital of Ningbo Medical Center from January 2019 to March 2020. By fiberoptic endoscopic evaluation of swallowing (FEES) combined with Penetration and Aspiration Scale (PAS), prospectively observed the aspiration and invasion of solid, paste, fluid and other food in the early stage of trial feeding in patients with laryngeal hypopharyngeal cancer undergoing open laryngeal function preservation.Results:Among the 69 patients, 21 had vertical partial laryngectomy and 19 had partial pharyngeal partial laryngectomy. 15 days after the operation, 2 groups of patients after solid and paste food adaptability scored (1.14±0.36), (1.29±0.56) and (2.53±2.04), (2.84±2.31) points, Friedman Mtest for comparison difference had statistical significance ( Mvalues were 23.463 and 22.227, P<0.01); the liquid food for adaptability scored (2.10±1.09), (4.42±2.24) points, the pairwise comparison of liquid, solid and paste showed statistically significant differences ( tvalues were-0.976 to 1.105, P<0.05). The pairwise comparison of the adaptability of the two groups on 20 days after surgery showed no statistically significant difference ( P>0.05).Throat on the cricoid cartilage resection-ring hyoid epiglottis anastomosis (SLCP-CHEP) was 17 cases, laryngeal glottis level partial resection was 12 cases; postoperative 15 days the adaptability on the behavior of three kinds of food grade 2 groups of patients (4.65±1.90), (5.59±1.46), (6.53±1.13) points and (6.67±1.07), (4.50±2.07), (6.92±0.79) points, respectively; Minspection differences were statistically significant ( Mvalues were 29.525, 22.136, P<0.01).The pairwise comparison of solid and paste food in the two groups 20 days after the operation showed no statistically significant difference ( P>0.05), while the difference of liquid and paste was statistically significant ( tvalues were-1.375 to-0.853, P<0.05). Conclusion:In the early recovery of patients undergoing laryngeal function preservation surgery, the vertical group and the partial pharyngeal partial laryngectomy group has better adaptability to solid and paste food. The horizontal group has better adaptability to paste, the CHEP group has better adaptability to solid, and the four groups has the worst adaptability to convection, the recovery time of convective mass between the CHEP group and the horizontal group was longer. To understand the adaptability of patients with different surgical methods to food traits at the early stage of trial feeding can help to implement targeted rehabilitation programs, carry out progressive diet training, and reduce the complications of misinvasion, aspiration, aspiration pneumonia and other complications in the recovery cycle.

Based on single-cell sequencing of the hippocampi of 5x familiar Alzheimer's disease(5x FAD)and wild type mice at 2-,12-,and 24-month of age,we found an increased percentage of microglia in aging and Alzheimer's disease(AD)mice.Blood brain barrier injury may also have contributed to this increase.Immune regulation by microglia plays a major role in the progression of aging and AD,according to the functions of 41 intersecting differentially expressed genes in microglia.Signaling crosstalk between C-C motif chemokine ligand(CCL)and major histocompatibility complex-1 bridges intercellular communi-cation in the hippocampus during aging and AD.The amyloid precursor protein(APP)and colony stimulating factor(CSF)signals drive 5x FAD to deviate from aging track to AD occurrence among intercellular communication in hippocampus.Microglia are involved in the progression of aging and AD can be divided into 10 functional types.The strength of the interaction among microglial subtypes weakened with aging,and the CCL and CSF signaling pathways were the fundamental bridge of communication among microglial subtypes.

There remains a significant gap in our quantitative understanding of crosstalk between apoptosis and necroptosis pathways. By employing the SWATH-MS technique, we quantified absolute amounts of up to thousands of proteins in dynamic assembling/de-assembling of TNF signaling complexes. Combining SWATH-MS-based network modeling and experimental validation, we found that when RIP1 level is below ~1000 molecules/cell (mpc), the cell solely undergoes TRADD-dependent apoptosis. When RIP1 is above ~1000 mpc, pro-caspase-8 and RIP3 are recruited to necrosome respectively with linear and nonlinear dependence on RIP1 amount, which well explains the co-occurrence of apoptosis and necroptosis and the paradoxical observations that RIP1 is required for necroptosis but its increase down-regulates necroptosis. Higher amount of RIP1 (>~46,000 mpc) suppresses apoptosis, leading to necroptosis alone. The relation between RIP1 level and occurrence of necroptosis or total cell death is biphasic. Our study provides a resource for encoding the complexity of TNF signaling and a quantitative picture how distinct dynamic interplay among proteins function as basis sets in signaling complexes, enabling RIP1 to play diverse roles in governing cell fate decisions.
Animals ; Apoptosis ; Caspase 8/metabolism* ; GTPase-Activating Proteins/metabolism* ; HEK293 Cells ; Humans ; Mice ; Mice, Knockout ; Necroptosis ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism*