Effects of high hydrostatic pressure on peelability and quality of crayfish(Procambarus clarkii).

BACKGROUND: Peeling of crayfish is a very important process in production. Crayfish peeling by machine can increase production efficiency and enhance safety in the production process. The tight muscle-shell attachment causes difficulty in peeling freshly caught crayfish. However, few studies have explored the changes in crayfish quality under favorable shell-loosening treatments. RESULTS: In this study, the shell-loosening properties of crayfish and changes in crayfish quality, microstructure and protein fluorescent features were investigated after high hydrostatic pressure (HHP) treatment. New methods were established to measure the peeling performance of crayfish, which are peelability and meat yield rate (MYR). The normalization of peelability and MYR were verified by different weights of crayfish tails and different treatments. The peeling effect of HHP-treated crayfish was evaluated by a new quantitative measurement method, and MYR was calculated. The results showed that all the HHP treatments reduced crayfish peeling work and increased MYR. The HHP treatment provided better crayfish quality in terms of texture and color and enlarged the shell-loosening gap. Among all HHP treatments, 200 MPa treatment exhibited lower peeling work, higher MYR and an expansion of the shell-loosening gap, reaching up to 573.8 µm. At the same time, 200 MPa treatment could maintain crayfish quality. CONCLUSION: The findings outlined above suggest that high pressure is a promising method for loosening crayfish shells. 200 MPa is an optimal HHP treatment condition for crayfish peeling, exhibiting a promising application in industrial processing. © 2023 Society of Chemical Industry.

Prevalence, characteristics, and distribution of HPV genotypes in women from Zhejiang Province, 2016-2020.

OBJECTIVE: To evaluate and understand the prevalence of HPV genotypes and characteristics of female populations in specific areas and the relationship with cervical lesions, which can effectively guide cervical cancer screening and formulate HPV vaccine prevention strategies. METHODS: A total of 77,443 women who visited gynecological clinics and underwent health examinations in the Second Affiliated Hospital of Zhejiang University School of Medicine during 2016-2020 were enrolled in this survey. Cervical samples were collected for HPV DNA genotyping and cervical cytology testing. Cervical biopsies were performed for patients with visible cervical abnormality or abnormal cytological results. RESULTS: The results showed the 5-year overall positive rate was 22.3%, of which the gynecology clinic group had significantly more positive results compared with the health examination group (P < 0.001). The five most common genotypes in Zhejiang Province were HPV 52, 58, CP8304, 16, and 51 (23.9%, 12.7%, 11.7%, 11.7% and 9.3%). HPV infection was age-specific, with the highest infection rate in the age group ≤ 20 compared to other age groups (P < 0.001). HPV infection was also season-specific, with the highest infection rate in spring or winter. The main HPV infection mode was single infection (P = 0.004), but patients ≤ 20 years old were more likely to develop multiple infections (51.0%). HPV 16, 52 and 58 were the main genotypes that caused cytological abnormalities and HPV16, 18, 56, 58 and 66 were independent risk factors for cervical lesions (OR = 2.352, 1.567, 2.000, 1.694, 1.889; all P < 0.05). Further analysis found HPV 16 and 18 were the main genotypes that cause cervical cancer histological abnormalities and were independent risk factors for cervical cancer (OR = 5.647, P < 0.001; OR = 3.495, P = 0.036). CONCLUSION: This article analyzed the prevalence of distribution characteristics of HPV infection and revealed the corelation between HPV infection and cytological and histological abnormalities. Comprehensive results of this survey will help Zhejiang Province to formulate public health policies and provide evidence for future selection of specific HPV vaccines.

Application of CRISPR/Cas9-Based Genome Editing in Studying the Mechanism of Pandrug Resistance in Klebsiella pneumoniae.

In this study, a CRISPR/Cas9-mediated genome editing method was used to study the functions of the mgrB, tetA, and ramR genes in mediating colistin and tigecycline resistance in carbapenem-resistant Klebsiella pneumoniae (CRKP). Inactivation of the tetA or ramR gene or the mgrB gene by CRISPR/Cas9 affected bacterial susceptibility to tigecycline or colistin, respectively. This study proved that the CRISPR/Cas9-based genome editing method could be effectively applied to K. pneumoniae and should be further utilized for genetic characterization.

MicroRNA Expression Profiling of Pancreatic Cancer Cell Line L3.6p1 Following B7-H4 Knockdown.

BACKGROUND/AIMS: Co-stimulating molecule B7-H4 regulates T cell-mediated immune responses, participates in tumor immune escape, and promotes the proliferation and metastasis of pancreatic cancer cells. However, the specific mechanisms are unclear. MicroRNAs (miRNAs) participated in the pathogenesis and progression of cancer. METHODS: In this study, a microarray technique was used to screen B7-H4-related differentially expressed miRNAs in a pancreatic cancer cell line find those associated with pancreatic cancer. Using a miRCURYTM LNA Array approach, we compared the miRNA expression profiles of L3.6p1 pancreatic cancer cells transfected with B7-H4 siRNA for 72 h with those transfected with non-target siRNAs. RESULTS: B7-H4 siRNA significantly up-regulated 57 miRNAs and down-regulated 14 miRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway analysis of predicted miRNA targets showed that these genes were mainly involved in protein binding, pathways in cancer, mitogen-activated protein kinase (MAPK) signaling pathway, and phosphatidylinositol 3-kinase-Akt (PI3K-Akt) signaling pathway. CONCLUSIONS: This is the first description of target genes of B7-H4, showing that miRNAs participate in the B7-H4 mediated regulation of oncogenicity and pathogenesis of pancreatic cancer. These results may help us better understand the role of B7-H4 in the progression of pancreatic cancer and its possible mechanisms. We also provide novel biomarkers for potential treatments of pancreatic cancer.

Rumen Microbiota Transplantation Alleviates Gossypol Diet-Induced Reproductive, Liver, and Intestinal Damage in Male Mice.

Ruminants exhibit stronger tolerance to gossypol, an anti-nutritional factor, compared to monogastric animals. We transplanted Hu sheep rumen microbiota into male mice to investigate the role of rumen microbiota in animal gossypol tolerance. Thirty specific-pathogen-free (SPF) male C57BL/6 mice were randomly divided into three groups: normal diet (CK group), gossypol diet (FG group), and rumen microbiota transplantation (FMT group, gossypol diet). The pathological changes in the liver and small intestine of the mice, the organ coefficient, and sperm parameters were analyzed. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the blood and lactate dihydrogen-X (LDH-X) levels in the testicular tissue were also measured. The results showed that body weight, feed intake, sperm concentration, sperm motility, and LDH-X levels in the FMT group increased (p < 0.05) compared with the FG group, while the enzyme activities of ALT, AST, and AST/ALT decreased (p < 0.05). In the FMT group, the injury to liver cells was alleviated, the structure of the small intestine was intact, and the villus height and the ratio of villus height to crypt depth (V/C) were higher than those in the FG group (p < 0.05). And there were no differences in various organ coefficients and sperm deformity rates among the three groups (p > 0.05), but compared with the FG group, mice in the FMT group showed tendencies closer to those in the CK group. Rumen microbiota transplantation relieved the reproductive toxicity and liver damage induced by gossypol in male mice and improved the tolerance of recipient animals to gossypol. Additionally, rumen microbes improved the intestinal structural integrity of recipients.

Effect of light treatmeat on oxidation and flavour of dry-cured Wuchang fish.

Lighting conditions are an important factor affecting dry-cured products. This study investigated the effects of treatments with different light intensities (0 lx, 1000 lx, 25000 lx) and different light sources including red light, blue light, UV-light on oxidation leve and flavor change in dry-cured Wuchang fish. The results showed that dry-cured Wuchang fish exhibited an attractive brown-yellow color, the highest oxidation degree of myoglobin (Mb), the highest fat oxidation under the light conditions of 25000 lx light intensity and UV-light irradiation. This phenomenon was observed that the degree of Mb oxidation was increased, while the degree of fat oxidation was increased. At 25000 lx light intensity and UV-light irradiation, dry-cured Wuchang fish showed an ignificantly decreased fatty acid conten (especially oleic acid and linoleic acid), significantly increased characteristic volatile compound contents (22 for 25,000 lx light intensity and 27 for UV-light irradiation), which contributed to the improvement of quality stability of dry-cured Wuchang fish. Our findings provide theoretical support for the industrial application of exogenous light in dry-cured Wuchang fish.

S100A6 is a potential diagnostic and prognostic biomarker for human glioma.

S100 calcium-binding protein A6 (S100A6) is a protein that belongs to the S100 family. The present study aimed to investigate the function of S100A6 in the diagnosis and survival prediction of glioma and elucidated the potential processes affecting glioma development. The Cancer Genome Atlas database was searched to identify the relationship among S100A6 expression, immune cell infiltration, clinicopathological parameters and glioma prognosis. Several clinical cases were used to verify these findings. S100A6 gene expression was high in glioma tissues, suggesting its diagnostic significance. In particular, S100A6 upregulation in glioma tissues exhibited a significant and positive correlation with the World Health Organization (WHO) grade, histological type, age, sex, primary treatment outcomes, 1p/19q codeletion, isocitrate dehydrogenase (IDH) status, overall survival (OS), progression-free interval and disease-specific survival. Kaplan-Meier and Cox regression analyses revealed that S100A6 gene expression can independently function as a risk factor affecting the prognosis of patients with glioma. Furthermore, Gene Ontology functional enrichment analysis revealed that S100A6 is implicated in immune responses and that the expression profiles of S100A6 are linked to the immune microenvironment. Furthermore, immunohistochemistry revealed that increased S100A6 protein levels are correlated with age, 1p/19q codeletion, IDH status, WHO grade and OS. The present findings suggest that increased S100A6 expression is an indicator of the dismal prognosis of patients with glioma and that it can be used as a potential diagnostic biomarker for this condition.

Microbial Evaluation of Ozone Water Combined with Ultrasound Cleaning on Crayfish (Procambarus clarkii).

The effects of ozone water (OW) and ultrasound cleaning (UL) on microbial community diversity of crayfish were studied through microbial viable count and 16S rRNA gene sequencing. The results showed that compared with the control (CK), the ozone water combined with ultrasound cleaning (OCU) showed a significant reduction (p < 0.05) in total viable count (TVC), psychrophilic viable count (PVC), mesophilic viable count (MVC), Pseudomonas, hydrogen sulfide-producing bacteria (HSPB), molds and yeasts. Concretely, the TVC of the CK, OW, UL and OCU were 5.09, 4.55, 4.32 and 4.06 log CFU/g, respectively. The dominant bacterium in untreated crayfish was Chryseobacterium, and its relative abundance was reduced by combined treatment. Color measurement and sensory evaluation suggested that a satisfactory sensory experience could be obtained on the crayfish applied with OCU. In brief, OCU could be used as a cleaning strategy to control the microbial quality of crayfish and have no influence on its quality.

Evaluation of qSOFA score, and conjugated bilirubin and creatinine levels for predicting 28-day mortality in patients with sepsis.

Sepsis is a dangerous disease that develops rapidly and has a high mortality rate. A timely and accurate assessment of the patient's condition is beneficial in improving prognosis and reducing mortality. Therefore, the present study was designed to investigate the potential association between quick sequential organ failure assessment (qSOFA) scores and biochemical indicators, such as conjugated bilirubin (CB) and creatinine levels, with the 28-day prognosis of patients with sepsis in a retrospective observational study. All cases were divided into survival and non-survival groups on the 28th day after diagnosis. The qSOFA scores, and CB and creatinine levels were significantly higher in the non-survival group than in the survival group (both P<0.01). Cox regression models identified CB [hazard ratio (HR), 1.006; P=0.002] and creatinine levels (HR, 1.002; P=0.024) as independent factors affecting 28-day mortality. The area under the curve (AUC) for CB and creatinine levels plus qSOFA score was 0.792 (95% confidence interval, 0.745-0.834), which was larger than the values for CB level, creatinine level and qSOFA score alone (all P<0.01) in the prognosis of 28-day mortality. The cut-off value of CB and creatinine levels plus qSOFA score for the 28-day mortality was 0.275 (-2.466 + 0.012 x CB + 0.002 x creatinine + 1.289 x qSOFA). Patients with lower combined predictor values had a better prognosis as demonstrated by Kaplan-Meier survival curves (log-rank test, 10.060; P=0.002). In both the septic shock and sepsis groups, the combined predictor value was higher in the non-survival group than in the survival group (P<0.001). Therefore, an increase in the combined predictor value of CB and creatinine levels plus qSOFA score may be an important predictor of disease progression and prognosis in patients with sepsis and septic shock.

Clinical Metagenomic Next-Generation Sequencing for Diagnosis of Secondary Glaucoma in Patients With Cytomegalovirus-Induced Corneal Endotheliitis.

Glaucoma is the second leading cause of blindness globally. Growing scientific evidence indicated that inflammation of the trabecular meshwork induced by corneal endotheliitis could lead to secondary glaucoma. Cytomegalovirus (CMV) has been identified as the most common herpes virus in corneal endotheliitis patients. Early detection is critical in preventing endothelial cell loss, and patient management should vary based on different pathological factors. However, routine culture and real-time polymerase chain reaction (qPCR) have difficult in distinguishing whether CMV, Varicella Zoster Virus (VZV) or Herpes Simplex Virus (HSV) causes endothiliitis. This may result in inappropriate treatment, which may prolong or aggravate the status of disease. We compared the sensitivity and specificity of qPCR and Metagenomic Next-Generation Sequencing (mNGS) in the aqueous humor of patients with suspected CMV endotheliitis in this study. Our results showed that four out of 11 (36.4%) of our patients were positive for CMV by qPCR, whereas mNGS had a 100% detection rate of CMV. Our findings implied that mNGS could be a useful diagnostic tool for CMV-induced endotheliitis.

B7-H4 is a prognostic biomarker for poor survival in patients with pancreatic cancer.

B7-H4 belongs to the immune costimulatory B7 family and is thought to negatively regulate T-cell-mediated immunity, and may contribute an important role in tumor immune evasion. Although the expression of B7-H4 has been observed in human pancreatic cancer, the prognostic significance of this expression is poorly understood. This present study explored the prognostic value of B7-H4 in pancreatic cancer. Patients with pancreatic cancer and healthy controls were recruited at the Second Affiliated Hospital to Zhejiang University from January 2011 to December 2014. Expression of B7-H4 was assessed by immunohistochemistry. Immunohistochemical analysis indicated that B7-H4 was expressed in 100% (188/188) of the pancreatic cancer tumor tissue samples, while only in 68% (17/25) of normal pancreatic tissue samples. Furthermore, the expression levels of B7-H4 in pancreatic cancer patients were significantly higher than in controls (P<.01). A significant difference in B7-H4 expression was observed between patients with late tumor-node-metastasis (TNM) stage (III and IV) and early TNM stage (I and II) (P<.01). The expression of B7-H4 was associated with distant metastasis (P<.01) and differentiation (P<.01). In addition, B7-H4 expression (P<.01), distant metastasis (P<.01), TNM stage (P<.01), differentiation (P<.01) and chemotherapy treatment (P<.05) were indicators of poor overall survival time. Multivariate survival analysis indicated that B7-H4 expression, distant metastasis, and chemotherapy treatment (P<.05) were independent prognostic indicators of poor overall survival. In conclusion, B7-H4 is highly expressed in pancreatic cancer, and is an independent predictor of poor prognosis in patients with pancreatic cancer. B7-H4 may represent an immunotherapeutic target in pancreatic cancer.

Effects of vanadate on the activities of mice glucokinase and hexokinase.

This study aimed at acquiring knowledge on the hypoglycemic mechanisms of sodium metavanadate (SMV) showed that the liver glucokinase and muscle hexokinase activities increased rapidly after oral SMV was given, and that the blood glucose level was correlated closely with the activities of the two enzymes but not with the insulin level; which indicated that SMV could improve the altered glucose phosphorylation in diabetic mice independently of stimulating insulin secretion. This was probably one of the mechanisms of hypoglycemic effects of SMV.

[Hypoglycemic effects of sodium metavanadate in diabetic mice and its effect on glucose phosphorylation].

OBJECTIVE: To investigate the effects of sodium metavanadate (SMV) on blood sugar and glucose phosphorylation in mice, and to discuss the possible mechanism of its hypoglycemic effects. METHODS: Diabetic mice (D) and control mice (V) were randomly allocated to drink SMV (0.2 mg/ml) (CV and DV groups) or NaCl (80 mmol/L) (C and V groups) respectively. The study lasted for 5 weeks. Liver glucokinase, muscle hexokinase, blood glucose and insulin were assayed at the end of each week. RESULTS: Blood glucose was higher in the diabetic groups before the administration of SMV, and the blood glucose level of group DV decreased from (18.77 +/- 1.28) to (8.94 +/- 0.94) mmol/L (P < 0.01) after oral administration of SMV for one week. While liver glucokinase increased from (1.29 +/- 0.64) to (15.36 +/- 1.57) mIU/min/mg protein and muscle hexokinase increased from (1.93 +/- 0.50) to (18.62 +/- 1.71) mIU/min/mg protein (P < 0.01) respectively. There was no continuous change of these parameters during the later weeks. No significant change of serum insulin was observed in the diabetic mice. There was a remarkable negative correlation of blood glucose level with liver glucokinase and muscle hexokinase levels. CONCLUSION: The hypoglycemic effects of SMV was independent of insulin level. In consideration of the close relations of the activities of liver glucokinase and muscle hexokinase with diabetes, and the improving of impaired glucose phosphorylation in diabetic mice by oral sodium metavanadate, which might be the mechanism of hypoglycemic effects of SMV.