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Abstract: An 8‐week feeding trial was conducted to evaluate the effects of fish meal (FM) replacement by rice protein concentrate (RPC) with supplementation of microcapsule lysine (ML) or crystalline lysine (CL) on growth performance, muscle development and flesh quality of blunt snout bream. Four isonitrogenous and isoenergetic diets were formulated, including FM diet (containing 50 g/kg FM), RPC diet (FM replaced by RPC), MRPC diet (FM replaced by RPC with ML supplementation) and CRPC diet (FM replaced by RPC with CL supplementation). Fish fed FM diet had significantly higher weight gain, feed efficiency, protein efficiency ratio and nitrogen and energy utilization than that of RPC group, but showed no statistical difference with other treatments. In addition, fish fed RPC diet showed higher muscle fibre frequency in the 20‐ to 50‐μm class but lower >50‐μm class and higher cooking loss than that of the other groups. Furthermore, no significant difference was found in whole‐body proximate compositions, frequency distribution of <20‐μm‐diameter fibres, texture, muscle content, collagen, pH 24 hr post‐mortem and sensory quality. The results showed that RPC supplemented with ML or CL could replace fishmeal without any adverse effects on growth performance and flesh quality for blunt snout bream. [ABSTRACT FROM AUTHOR]

Abstract: Five 2‐week feeding trials were conducted to investigate five stimulants on P. sinensis. Two isonitrogenous and isoenergetic diets were formulated, one contained 600 g kg–1 fishmeal (FM diet) and the other contained 420 g kg–1 fishmeal and 225 g kg–1 animal protein blend (APB diet), and each tested one stimulant with four levels. The feeding stimulant candidates were betaine, 2‐carboxy‐ethyl dimethyl sulphonium bromide (DMPT), inosine‐5′‐monophosphate (IMP), taurine and squid extract. In the experiment on each stimulant, turtles (10.85 ± 0.10 g) were equally divided into FM and APB groups and fed with an equal mixture of corresponding diets containing four levels of stimulants, respectively. Each diet contained a unique rare earth oxide as inert marker. Turtles were fed twice daily (8:00 and 17:00), and faeces were collected. Preference for each diet was estimated based on the relative concentration of each marker in the faeces. In the FM group, only IMP showed the enhanced attraction. But in APB groups, all the stimulants, except IMP, showed higher preference than basal diets for at least one inclusion level, and the optimum level was 10 g kg–1 for betaine, 0.1 g kg–1 for DMPT, 0.1 g kg–1 for IMP, 5 g kg–1 for taurine and 10 g kg–1 for squid extract, and the squid extract had the strongest stimulating effect among the stimulants. [ABSTRACT FROM AUTHOR]

This study aimed to determine the optimal dietary niacin requirement of juvenile Megalobrama amblycephala. Fish, with an average weight of 3.62 ± 0.02 g, were randomly divided into six groups and fed six purified diets with graded levels of niacin (0, 10, 20, 30, 40 and 50 mg/kg) three times daily for 8 weeks. The survival rate, weight gain ( WG), feed conversion ratio, protein efficiency and protein retention all significantly improved ( p < .05) as the dietary niacin level increased from 0 to 30 mg/kg but showed no significant difference ( p > .05) with further increases in niacin levels. The supplementation of 30 mg/kg niacin also led to a significant ( p < .01) reduction in the viscerosomatic index, hepatosomatic index and mesenteric fat index. The dietary niacin levels exhibited minimal effects on whole-body moisture and ash content ( p > .05) but exerted a significant ( p < .05) influence on protein and lipid contents and liver nicotinamide concentrations with high values obtained in fish receiving greater than 30 mg/kg of niacin. In addition, moderate levels of niacin lowered plasma triglycerides, non-esterified fatty acid and low-density lipoprotein cholesterol concentrations ( p < .05). Based on the broken-line regression analysis of WG and liver nicotinamide content, the optimal dietary niacin requirement of juvenile blunt snout bream was 31.25 and 30.62 mg/kg, respectively. [ABSTRACT FROM AUTHOR]

An 8-week feeding trial was performed to investigate effects of dietary selenium (sodium selenite, Na2SeO3; selenium nanoparticle, Nano-Se; selenium yeast, Se-yeast) on the growth, selenium status, antioxidant activities, muscle composition and meat quality of blunt snout bream. Na2SeO3 and Nano-Se were supplemented at 0.2 mg Se kg−1, and Se-yeast was supplemented at 0.1, 0.2, 0.4 and 0.8 mg Se kg−1 in basal diet; no Se was added as a control. The results indicated that groups of 0.2 and 0.4 mg kg−1 Se-yeast had significantly higher weight gain and Nano-Se, 0.2, 0.4 mg kg−1 Se-yeast had significantly lower feed conversation ratio compared with the control group. The Se concentrations of whole body, muscle and liver linearly increased with increasing dietary Se-yeast levels. Group of 0.4 mg kg−1 Se-yeast significantly increased activities of catalase and glutathione peroxidase. Muscle colour of Nano-Se, 0.2, 0.4 mg kg−1 Se-yeast groups, the water-holding capacity of 0.4, 0.8 mg kg−1 Se-yeast groups showed significant differences compared with the control group. The results suggest that Se-yeast and Nano-Se had a better growth performance than Na2SeO3 at 0.2 mg Se kg−1, and supplementing appropriate Se-yeast in diet can increase antioxidant activities and enhance meat quality of blunt snout bream. [ABSTRACT FROM AUTHOR]

An 8-week feeding trial was conducted to evaluate the potential of partial replacement of fish meal with rendered animal protein blend ( APB) [meat and bone meal (MBM): expanding blood meal (EBM) = 4 : 1] in the diet of juvenile Chinese soft-shelled turtle Pelodiscus sinensis. Seven isonitrogenous and isoenergetic practical diets replacing 0%, 10%, 20%, 30%, 40%, 50% and 60% fish meal protein by APB were formulated. Weight gain, feed efficiency, protein efficiency and the apparent digestibility coefficients ( ADCs) of dry matter and gross energy in the experimental diets all exhibited no significant difference ( P > 0.05) when dietary fish meal was replaced for 0-40%, but decreased significantly ( P < 0.05) with further increasing substitution levels. However, relative feed intake, ADC of lipid as well as the contents of whole-body moisture, protein, lipid and ash all showed little differences ( P > 0.05) among all the treatments. Nitrogen retention and the ADCs of protein and most amino acids all decreased significantly ( P < 0.05) when fish meal was substituted by more than 30%. The results indicated that fish meal in the practical diet of juvenile Chinese soft-shelled turtle could be replaced by 30-40% using a combination of MBM and EBM without negative effects on growth performance, nutrients digestibility and body composition. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the optimal dietary biotin requirement of juvenile Megalobrama amblycephala. Quadruple groups of fish (initial average weight 2.01 ± 0.01 g) were fed thrice daily with six isonitrogenous and isoenergetic purified diets containing 0 (basal diet), 0.015, 0.049, 0.158, 0.624 and 2.49 mg kg−1 biotin, respectively, for 8 weeks. Results showed that survival rate, final weight, feed intake, specific growth rate, protein efficiency ratio and nitrogen retention efficiency all increased significantly ( P < 0.01) as dietary biotin levels increased from 0 to 0.049 mg kg−1, whereas the opposite was true for feed conversion ratio. Dressout percentage, condition factor, hepatosomatic index, viscera/body ratio all showed no significant difference ( P > 0.05) within the biotin range tested. Contrary to moisture content, whole-body protein and lipid contents showed a positive correlation with dietary biotin levels. In addition, liver biotin content increased significantly ( P < 0.05) with increasing dietary biotin levels up to 0.624 mg kg−1. Hepatic pyruvate carboxylase ( PC) and acetyl- Co A carboxylase ( ACC) activities both showed an increasing trend as dietary biotin levels increased. Based on the regression analysis of weight gain, hepatic PC and ACC activities, the optimal dietary biotin requirement of juvenile Megalobrama amblycephala is estimated to be 0.063, 0.071 and 0.075 mg kg−1, respectively. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the optimum dietary carbohydrate/lipid ( CHO/L) ratio for fingerling blunt snout bream Megalobrama amblycephala (average initial weight: 6.61 ± 0.03 g). Six isonitrogenous and isoenergetic diets were formulated to contain various CHO/L ratios ranging from 1.62 to 24.20. Each diet was tested in four replicates for 10 weeks. Specific growth rate ( SGR), feed conversion ratio and nitrogen and energy retention all improved significantly ( P < 0.05) as dietary CHO/L ratios decreased from 24.20 to 5.64 but showed little difference ( P > 0.05) as dietary CHO/L ratios ranged from 2.45 to 5.64. Hepatosomatic index increased significantly ( P < 0.05) as dietary CHO/L ratios increased, whereas intraperitoneal fat ratio showed an opposite trend ( P < 0.05). Opposite to moisture content, lipid content of whole body and carcass all increased significantly ( P < 0.05) as dietary CHO/L ratios decreased. Liver lipid content showed little difference ( P > 0.05) among all the treatments, while liver glycogen content increased significantly ( P < 0.05) with increasing CHO/L ratios. High dietary carbohydrate enhanced the activities of liver hexokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase but did not induce hyperglycaemia. Based on the second-order polynomial regression analysis of SGR, the optimal dietary carbohydrate and lipid contents for fingerling blunt snout bream were 291.7 and 81.4 g kg−1, respectively, with a corresponding dietary CHO/L ratio of 3.58. [ABSTRACT FROM AUTHOR]

A 12-week feeding trial was conducted to determine dietary choline requirement for juvenile Megalobrama amblycephala. The basal diet was formulated to contain 310 g kg−1 diet from vitamin-free casein and gelatine. Choline chloride was supplemented to the basal diet to formulate six purified diets containing 0, 250, 500, 1000, 2000 and 4000 mg kg−1, respectively. Each diet was randomly fed to quadrupled groups of Megalobrama amblycephala with initial average weight 1.84 ± 0.04 g in a flow-through system. Results showed weight gain was increased significantly with increasing dietary choline levels ( P < 0.01). Lipid content of liver decreased significantly as dietary choline concentration increased ( P < 0.01), whereas lipid content of dressed carcass showed opposite trend ( P < 0.01), and lipid content of whole-body was unaffected by dietary choline supplementation. Broken- line regression of weight gain, liver and muscle choline concentration showed choline requirements of Megalobrama amblycephala of 1198, 1525 and 1365 mg kg−1, respectively. In addition, dietary choline supplementation significantly improved lipid content of dressed carcass but not the content of whole body of blunt snout bream. [ABSTRACT FROM AUTHOR]

Objective: To explore the clinicopathological features, immunophenotype and molecular genetic characteristics of malignant solitary fibrous tumor (MSFT). Methods: Seven cases of MSFT were collected from the First Affiliated Hospital of Zhengzhou University from July 2018 to December 2020. Immunohistochemistry, RNA-based NGS and DNA-based NGS were performed. Result s Among the 7 patients, there were 5 males and 2 females with a median age of 53 years (37-69 years). Two tumors located at skull base, and one in the tentorium of cerebellum, parietal occipital region, occipital area, chest and buttock respectively. The maximum diameter of the tumor was 2.5-20.0 cm. Microscopically, typical hemangiopericomatoid structures were noted; the tumor was cellular, fusiform or oval, very pleomorphic, with necrosis and high mitotic figures (>4/10 HPF). In some cases, classical solitary fibrous tumor morphology and dedifferentiated region were observed. Immunohistochemically, the tumor was positive for CD34 (6/7), STAT6 (7/7), bcl-2 (7/7), but negative for S-100 (7/7); CKpan or EMA was positive to varying degrees; mutated p53 was noted (3/7); Ki-67 positive index was more than 10%. NAB2-STAT6 gene fusion was typically detected in all the 7 cases. In 4 cases, ZNF415-FGFR1, COPG1-MET, IPO11-LRRC70&#95;ncRNA-PLAG1 and Clorf198-CD274 (PD-L1) gene fusions were also detected. NOTCH1 mutation was found in 7 cases and TP53 mutation in 4 cases. TERT promoter mutations were not detected in all the cases. Conclusions: MSFT is rare and needs to be differentiated from many other spindle cell tumors. Especially when tumors express epithelial markers, they are easily misdiagnosed as sarcomatoid carcinoma and synovial sarcoma, etc. Immunohistochemistry and molecular detection of NAB2-STAT6 gene fusion have important diagnostic values. NOTCH1 and TP53 mutations may be associated with the progression of MSFT. Some patients have FGFR1 gene fusion and MET gene fusion, which may be potential therapeutic targets.

Objective: To investigate the impact and clinical significance of the revised 2019 Chinese HER-2 testing guidelines on the detecting result evaluation of invasive breast cancers with equivocal HER-2 immunostaining by using fluorescence in situ hybridization (FISH). Methods: A total of 569 cases of invasive breast cancers with HER-2 (+ + ) immunostaining evaluated according to the immunohistochemistry (IHC) guidelines of 2014 edition and 2019 edition from May to November 2019 were collected and further detected by FISH. The results of HER-2/CEPl7 double probe were respectively interpreted according to both the 2014 and 2019 Chinese HER-2 testing guidelines and the results were compared. Results: According to the 2014 guidelines, the number of HER-2 positive, equivocal and negative cases were 139 (24.43%), 67 (11.78%), and 363 (63.80%), respectively. Whereas according to the 2019 guidelines, 115 cases (20.21%) were the first group, 9 cases (1.58%) were the second group, 15 cases (2.64%) were the third group, 67 cases (11.78%) were the fourth group, and 363 cases were (63.80%) the fifth group, of which 130 cases (22.85%) were positive and 439 cases (77.15%) were negative by FISH detecting. Compared with the guideline of 2014 edition, the HER-2 positive rate of FISH detection reduced from 24.43% (139/569) to 22.85% (130/559) according to the application of the guideline of 2019 edition, but the difference was not statistically significant ( P =0.567), while the negative rate increased from 63.80% (363/569) to 77.15% (439/569), with a statistically significant difference ( P <0.05). Forty-three cases with incomplete weak to medium intensity of IHC membrane staining which were HER-2 (+ + ) according to 2014 guideline were changed to IHC (+ ) on the basis of the 2019 guideline. According to the FISH guideline of 2014 edition, 1 case (2.33%) was positive, 6 cases (13.95%) was equivocal and 36 cases (83.72%) was negative, while according to the 2019 FISH guideline, all of the 43 cases were negative. Conclusions: According to the guideline of 2019 edition, a proportion of cases changes from HER-2 (+ + ) to (+ ), and the HER-2 positive rate of FISH test decreases slightly, the negative rate increases, the equivocal result is eliminated, which provides a definite reference for screening patients who will be benefited from the targeted treatment of HER-2.

Objective: To find the biomarkers that accurately predict the survival of patients with esophageal squamous cell carcinoma (ESCC). Methods: The immune related genes that were significantly related to the overall survival (OS) of patients with ESCC were screened from The Cancer Genome Atlas (TCGA) database to construct a prognostic risk score model. The prognoses of the high-risk and low-risk groups were compared by Kaplan-Meier method. The accuracy of the model was evaluated by the receiver operating characteristic (ROC) curve. Tumor tissue samples of 83 patients with pathological diagnosis of ESCC were collected from Anyang Cancer Hospital for external verification. Cox regression analysis was used to comprehensively evaluate the effects of prognostic risk score and various clinical characteristics on OS of patients with ESCC. Results: Seven immune-related genes that were significantly related to survival prognosis were selected from the TCGA database and included in the prognostic risk score model, which were S100A12, SLC40A1, FABP9, TNFSF10, IGHA2, IL1F10, and STC2. The 1- and 2-year survival rates of the low-risk group (40 cases) were 94.3% and 82.5%, respectively, while those of the high-risk group (40 cases) were 75.9% and 32.9%, respectively.The prognosis of the high-risk group was worse than that of the low-risk group ( P <0.001). The 83 external validation samples obtained consistent results by using the prognostic risk score model. The prognostic risk score was positively correlated with the content of CD4(+) T lymphocytes in ESCC ( r ( s )=0.259, P =0.020), but not correlated with the content of B lymphocytes, CD8(+) T lymphocytes, neutrophils, macrophages or dendritic cells ( P >0.05). Conclusions: S100A12, SLC40A1, FABP9, TNFSF10, IGHA2, IL1F10, and STC2 were risk genes significantly associated with OS of patients with ESCC. The prognostic risk score was an independent prognostic factor for the OS of patients with ESCC, and it was correlated with the content of CD4(+) T lymphocytes in ESCC tissue.

Objective: To study the clinicopathological features and prognosis of nodal lymphoplasmacytic lymphoma/Waldenstrom's macroglobulinemia (n-LPL/WM). Methods: A total of 19 cases of n-LPL/WM were collected from May 2009 to January 2020 at First Affiliated Hospital of Zhengzhou University. The clinicopathologic features, immunophenotype, Ig gene rearrangement (BIOMED-2), MYD88 L265P mutation status (by Sanger sequencing) and follow-up data (by telephone) were analyzed. Results: There were 15 males and 4 females with a median age of 61 years (range 43 to 82 years). There were 14 WM and five LPL. The most common symptoms were weakness, fatigue (9/19) and B symptoms (11/19). Majority of the patients (16/18) presented with systemic multiple lymphadenopathies. Eighteen patients presented at advanced stages (Ⅲ/Ⅳ stage). Serum M protein status was IgM (15 cases), IgG (1 case), IgA (1 case) and no-secretory type (2 cases). Seventeen patients had bone marrow involvement. Morphologically, all 19 cases were divided into two groups: typical group (9 cases) or atypical group (10 cases). In the typical group, the structures of the lymph nodes were preserved; the neoplastic cells were predominantly plasmacytoid lymphocytes or mixed small lymphocytes, plasmacytoid lymphocytes and plasma cells, without proliferation of FDC network and follicular implantation. In the atypical group, the tumor showed effaced nodal architecture (5 cases), mainly proliferation of small lymphocytes (6 cases), FDC proliferation and/or follicular implantation (6 cases), marginal zone B cell differentiation (4 cases) and diffuse amyloidosis (1 case). Hemosiderin deposition (19 cases), infiltration of fatty tissue (19 cases) and interstitial sclerosis (9 cases) were commonly seen in both groups. Immunohistochemically, the neoplastic B cells expressed CD20 and CD79α, and the neoplastic plasma cells were positive for CD38, CD138 and MUM-1; eight cases showed light chain restriction; of the seven detected cases, five expressed IgM and the other two expressed IgG and IgA respectively; four cases expressed CD23 weakly, Ki-67 index was 10%-30%. MYD88 L265P mutation was seen in 18/18 cases. There was no significant difference in clinicopathologic features and prognosis between the two groups ( P >0.05). The median follow-up time was 61 months, 11 patients were alive, while eight died; the 5-year survival rate was 21.1%. Conclusions: n-LPL/WM is rare, but patients usually present in advanced stages. It is easily confused with other small B-cell lymphomas with plasma cell differentiation, especially basing on morphologic features alone; thus the accurate diagnosis of n-LPL/WM requires a combination of clinical features, serum M protein, immunohistochemistry, bone marrow morphology,flow cytometry and MYD88 L265P mutation status etc. The prognosis of n-LPL/WM may be not very good, and further studies with more cases are needed.

Objective: To study the clinicopathologic features and MYD88 L265P mutation status of intravascular large B cell lymphoma (IVLBCL). Methods: Fourteen cases of IVLBCLs were diagnosed from March 2014 to December 2019 at the First Affiliated Hospital of Zhengzhou University. The clinicopathologic features and prognosis were analyzed. Epstein-Barr virus encoded RNAs and MYD88 L265P mutation status were detected using in situ hybridization and Sanger sequencing, respectively. The follow-up data were obtained by telephone interview. Results: There were 6 males and 8 females with a median age of 62 years (range: 48-73 years). The involved anatomic locations were demonstrated by positron emission tomography-computed tomography, including adrenal gland (7/14), bone (6/14), central nerve system (4/14), skin (3/14), female reproductive system (3/14), local lymph nodes (3/14), prostate (2/14), liver and spleen (2/14), sphenoid sinus (1/14), penis (1/14), bladder (1/14), and right lung (1/14). Fever was the most common symptom (7/14), followed by neurologic symptoms and lower abdominal pain (2/14 each). The reminder symptoms included rash with edema, legs weakness and numbness, or postmenopausal bleeding (1/14 each). Eleven cases were at Lugano stage Ⅳ. Four cases were associated with the hemophagocytic syndrome, while 6 cases with bone marrow involved. Microscopically, the tumor cells were generally concentrated within the small-to-medium vascular lumens or sinusoids; they had centroblast-like appearance and showed large round or oval nuclei with slightly irregularities, coarse chromatin and 1-3 distinct nucleoli. One exception was the one case with an embryoid nuclei, reminiscent of anaplastic large cell lymphoma. The mitosis was not uncommon. Extravascular neoplastic cells were seen in two cases. The neutrophils could be appreciable in most of the cases (10/14). Immunophenotyping showed that CD20 and CD79α were diffusely and strongly positive in 14 cases; 12 cases were classified as the non-GCB subtype; 6 out of the 11 cases were double expressor lymphoma; 7 out of the 12 cases were CD5-positive. Twelve cases were EBER negative. The MYD88 L265P mutation was detected in 1 case (1/10). The duration of the follow-up ranged from 0.5 to 24.0 months, and 11 patients survived and 3 died. Conclusions: IVLBCL is rare. The most common type of IVLBCL in China is Asian type with scant tumor cells. Combination of clinical and immunohistochemical features can avoid most, if not all, misdiagnoses and missed diagnoses. Some IVLBCL cases may harbor the MYD88 L265P mutation, but the prevalence of MYD88 L265P mutation in the population still warrants additional studies.