[Artificial liver support system for acute-on-chronic liver failure combined with successful liver transplantation in stage III - IV hepatic encephalopathy: an analysis of 14 cases].

Objective: To evaluate the effect of plasma exchange combined with high-dose continuous venovenous hemodiafiltration method (CVVHDF) in the treatment of patients with acute-on-chronic liver failure with stage III-IV hepatic encephalopathy and the feasibility of pre-operative preparation for liver transplantation. Methods: Clinical data of 14 cases of medical intensive care unit of our hospital with acute-on- chronic liver failure accompanied with stage III-IV hepatic encephalopathy that underwent plasma exchange combined with high-dose CVVHDF from March 2015 to September 2017 were retrospectively summarized. The indexes of liver and kidney function, blood coagulation function, arterial blood PH, lactic acid and blood ammonia were monitored before and after treatment. Heart rate, blood pressure, APACHE II score, and consciousness recovery time were observed. Student's t- test was used to compare the mean values between the two groups. Results: Serum total bilirubin (t = 9.43, P < 0.01), serum creatinine (t = 3.40, P < 0.01), serum ammonia (t = 10.64, P < 0.01), prothrombin activity (t = 9.19, P < 0.01), serum lactate (t = 9.25, P < 0.01), heart rate (t = 4.47, P < 0.01), and mean arterial pressure (t = 4.41, P < 0.05) were significantly improved in 14 patients before and after treatment. In addition, respiratory rate (t = 6.01, P < 0.01) and APACHE II score (t = 7.19, P < 0.01) were significantly improved (P < 0.05). Eight patients with stage III hepatic encephalopathy were treated with intermittent plasma exchange combined with CVVHDF for 3 to 14 days, and six patients with stage IV were transformed to stage III to II. Liver transplantation was successfully performed on 14 patients with shortest time duration of 3days, and longest time duration of 1 month. Conclusion: Plasma exchange combined with CVVHDF can significantly improve liver and kidney functions, reduce blood ammonia level and improve mental health in patients with hepatic failure accompanied with stage III-IV hepatic coma. In addition, it also effectively increases the average arterial pressure, maintain stability of vital signs, maintain fluids, electrolytes and acid-base balance, create a stable internal environment for liver transplantation before operation, and extend time for liver transplantation.

Significant improvement of intestinal microbiota of gibel carp (Carassius auratus gibelio) after traditional Chinese medicine feeding.

AIMS: Increasing attention has been attracted to intestinal microbiota, due to interactions with nutrition, metabolism and immune defence of the host. Traditional Chinese medicine (TCM) feed additives have been applied in aquaculture to improve fish health, but the interaction with fish gut microbiota is still poorly understood. This study aimed to explore the effect of adding TCM in feed on the intestinal microbiota of gibel carp (Carassius auratus gibelio). METHODS AND RESULTS: Bacterial communities of 16 fish intestinal contents and one water sample were characterized by high-throughput sequencing and analysis of the V4-V5 region of the 16S rRNA gene. The results showed that the composition and structure of the bacterial community were significantly altered by the TCM feeding. Some phyla increased markedly (Proteobacteria, Actinobacteria, Acidobacteria, etc.), while Fusobacteria were significantly reduced. Concurrently, the richness and diversity of the taxonomic units increased, and the microbiota composition of TCM-treated fish was more homogeneous among individuals. At the genus level, the addition of TCM tended to reduce the incidence of potential pathogens (Aeromonas, Acinetobacter and Shewanella), while stimulating the emergence of some potential probiotics (Lactobacillus, Lactococcus, Bacillus and Pseudomonas). CONCLUSIONS: These data suggested that the feed additive could regulate the fish intestinal microbiota by reinforcing the microbial balance. SIGNIFICANCE AND IMPACT OF THE STUDY: This study may provide useful information for further application of TCM for diseases prevention and stress management in aquaculture.

Effect of skeletal muscle fibers on porcine meat quality at different stages of growth.

In this study, we examined changes in meat quality and content of muscle types during porcine growth. The influence of the longissimus dorsi muscle fiber composition on meat quality and the correlation between 2 fiber-typing methods (histochemistry and real-time quantitative polymerase chain reaction) were examined. Type IIx and type IIb fibers accounted for most of the total number of fibers; the proportion of these fibers increased during porcine growth (75.42, 80.09, and 79.88%, respectively, at 3 different stages of growth). There was a strong positive correlation between the 2 fiber-typing methods; the correlation coefficients of type I, IIa, and IIx+IIb fiber contents were 0.65, 0.88, and 0.92, respectively. The a* value of meat color was significantly lower at 98 days and negatively correlated with white fiber content (r = -0.69, P < 0.01). Water-holding capacity decreased during porcine growth. The drip loss parameter was positively correlated with type IIx+IIb fiber content (r = 0.55, P < 0.05). Decreased pH was strongly positively correlated with type IIx+IIb fiber content (r = 0.61, P < 0.01) and negatively correlated with type IIa fiber content (r = -0.44, P < 0.05). Therefore, we found that the composition of muscle fibers influenced the establishment of meat quality and its alteration during the early postmortem period.

Effect of ß-hydroxy-ß-methylbutyrate calcium on growth, blood parameters, and carcass qualities of broiler chickens.

Beta-hydroxy-ß-methylbutyrate (HMB), the metabolite of leucine, plays an important role in muscle protein metabolism. To investigate the effect of dietary HMB calcium (HMB-Ca) on growth performance, breast muscle development, and serum parameters in broiler chickens, a total of two hundred seventy 1-d-old Arbor Acres male broiler chicks were randomly allotted into 3 dietary treatments supplemented with 0, 0.05%, or 0.1% HMB-Ca during the starter (1 to 21 d) and grower (22 to 42 d) period. The results showed that broilers fed 0.1% HMB-Ca diet had higher ADG during the starter or the whole period, and gain 148 g more BW than the chicks fed the control diet at 42 d of age (P < 0.05). At 21 d of age, birds receiving 0.1% HMB-Ca had more breast muscle yield, less abdominal fat than the control, and more dressing percentage than birds fed the control or 0.05% HMB-Ca diet (P < 0.05). At 42 d of age, 0.1% HMB-Ca increased breast muscle yield than the control and decreased abdominal fat compared with the control or 0.05% HMB-Ca group (P < 0.05). In comparison with the control, feeding 0.1% HMB-Ca increased the triiodothyronine, thyroxine, triiodothyronine/thyroxine ratio and decreased the serum uric acid level at d 21 (P < 0.05). At 42 d of age, serum thyroxine level was elevated in the 0.05% HMB-Ca treatment, and the uric acid concentration was significantly decreased by the 0.1% HMB-Ca-supplemented diet (P < 0.05). Dietary HMB-Ca did not affect the growth hormone or insulin content. This study suggested that dietary supplementation of HMB-Ca improved growth performance, stimulated the breast muscle development, and decreased the abdominal fat deposition in broiler chickens, and the favorable effects were more pronounced in the starter phase. The growth promotion effect of HMB-Ca may be partly related to the increased serum thyroid hormones in broiler chickens.

Effects of constant and cyclic heat stress on muscle metabolism and meat quality of broiler breast fillet and thigh meat.

This study investigated the effects of constant and cyclic heat stress on muscle metabolism and meat quality of broiler breast fillet and thigh meat from 4 to 6 wk of age. Male Arbor Acres (AA) broilers (n = 270, 4 wk old) were raised under different temperature conditions: standard (temperature was 23°C); constant high temperature (temperature was 34°C); and cyclic high temperature (temperature was 36°C from 1000 h to 1600 h and 23°C from 1600 h to 1000 h). On d 42, broilers were stunned and sampled. The results showed that chronic high temperature significantly decreased the proportion of breast muscle and significantly increased the proportion of thigh muscle (P < 0.05). The moisture concentration was significantly higher in the breast muscle of the birds exposed to constant high temperature (P < 0.05), whereas the protein content was significantly lower (P < 0.05) and fat deposition was significantly higher (P < 0.05) in the breast muscle of the birds exposed to constant or diurnal cyclic high temperature than those grown under standard temperature. The breast and thigh muscle of the birds grown under constant high temperature had significantly higher lightness, cook loss, and shear force (P < 0.05) and significantly lower initial pH (pH(i)), ultimate pH (pH(u)), and redness compared with those grown under standard temperature (P < 0.05). The pH(i), pH(u), and redness were significantly lower (P < 0.05) while the lightness and shear force were significantly higher for the breast muscle of the chickens raised under diurnal cyclic high temperature (P < 0.05) than those grown under standard temperature. In contrast, lightness and yellowness of thigh muscle were significantly higher (P < 0.05) in the chickens grown under diurnal cyclic high temperature than under standard temperature. Breast and thigh muscle of broilers exposed to constant high temperature produced higher (P < 0.05) lactic acid and pyruvate kinase activities than those exposed to the standard temperature. These results indicated that chronic heat stress significantly increased lactate production, reduced meat pH value by accelerating meat glycolysis, and eventually reduced meat quality.

Silica Induces Plasminogen Activator Inhibitor-1 Expression through a MAPKs/AP-1-Dependent Mechanism in Human Lung Epithelial Cells.

ABSTRACT Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the silica-induced pulmonary fibrosis. The effect of silica on the expression of PAI-1 was investigated in human lung epithelial cells (A549). Silica induced PAI-1 expression in a concentration-(50-200 mug/mL) and time-(4-24 h) dependent manner in A549 cells. Furthermore, the roles of mitogen-activated protein kinase (MAPK)/activator protein-1 (AP-1) signaling pathways in silica-induced PAI-1 expression were examined. We found that silica (200 mug/mL) treatment for 4 to 24 h resulted in AP-1 activation in A549 cells. Cells were pretreated with the AP-1 inhibitor curcumin (10, 25, 50 muM), and silica-induced PAI-1 expression was reduced by 20%, 63%, and 65%, respectively. In addition, dominant-negative mutant c-Jun (TAM67) down-regulated silica-induced PAI-1 expression by 59%. P38 kinase inhibitor SB203580 (20 muM) and Erk inhibitor PD98059 (50 muM) suppressed silica-induced PAI-1 expression by 35% and 51%, respectively. Additionally, PD98059 but not SB203580 inhibited the AP-1 DNA binding activity induced by silica. The results suggest that the PAI-1 expression induced by silica may be involved in the activation of MAPKs/AP-1 signaling pathways in human lung epithelial cells.

[p16 and cyclinD1 protein expression and p16 gene mutation in primary human hepatocellular carcinoma].

OBJECTIVE: To elucidate the relationship between expression of p16 and cyclinD1 proteins and evaluate the role of p16 gene exon 2 mutation in hepatocellular carcinogenesis. METHODS: Streptavidin-peroxidase conjugated method(SP) was used to detect expressions of p16 and cyclinD1 proteins in 44 cases of hepatocellular carcinoma(HCC), and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) to detect p16 gene mutation in exon 2 in 12 cases of HCC and liver tissues adjacent to HCC. RESULTS: Expression rate and positive signal intensity of p16 protein in HCC were significantly lower(P < 0.01) and those of cyclinD1 protein in HCC were significantly higher (P < 0.05) than those in pericarcinomatous tissues. Of 12 fresh HCC tissues, p16 gene mutation in exon 2 was found in 2 cases, whereas that was not found in pericarcinomatous tissues. CONCLUSIONS: 1. Inactivation or/ and deletion of p16 protein may be one of important reasons which result in proliferation unbalance of cells. 2. p16 gene mutation in exon 2 presents in HCC, but it does not frequently occur in Chinese hepatocarcinogenesis. 3. p16 gene abnormality and cyclinD1 over expression may coact in HCC.

[Effect of activation of p-MAPK on activating c-fos and c-jun proteins in breast cancer].

OBJECTIVE: This paper was to investigate the relationship between expression of p-MAPK and oncogenesis of breast cancer. METHODS: Immunohistochemical technique was used to detect the expression of p-MAPK and c-fos and c-jun proteins in 68 cases of breast cancers, 42 cases of pericarcinomatous tissues and 7 cases of normal breast tissues. RESULTS: Positive stainings of p-MAPK, c-fos, and c-jun were localized in cancer cell nuclei. The positive rates of p-MAPK, c-fos, and c-jun were 86.8% (59/68), 82.4% (56/68), and 77.9% (53/68), respectively, which were much higher than that in pericarcinomatous tissues (P < 0.01). Of some cases, p-MAPK positive staining was also found in the nuclei of the fibroblastic and angioendothelial cells of cancer stroma. CONCLUSION: Activated or overexpressive MAPK activates the immediately-early oncogenes(c-fos, c-jun), which might play an important role in carcinogenesis of breast cancer and be an early event of oncogenesis of breast cancer.

[Role of functional inactivation of p53 from MDM2 overexpression in hepatocarcinogenesis].

OBJECTIVE: This paper was to evaluate the role of p53 mutation, and p53 and MDM2 proteins expression in hepatocarcinogenesis. METHODS: Using streptavidin-peroxidase conjugation method (SP), the expression of p53 and MDM2 proteins was observed in 61 cases of primary hepatocarcinomas (HCC) and 59 cases of corresponding paracancerous tissue, among which p53 mutations in exons 5-8 were detected in 21 cases by polymerase chain reaction single-strand confirmation polymorphism analysis (PCR-SSCP). RESULTS: Positive nuclear p53 and MDM2 immunostainings were demonstrated in 57.38% (35/61) and 26.23% (16/61) of HCC, and 1.69% (1/59) and 3.39% (2/59) of corresponding paracancerous tissue, respectively. The expressions of p53 and MDM2 proteins in HCCs were significantly higher than those in paracancerous tissues (P < 0.01). The expressions of p53 and MDM2 were not significantly correlated (P > 0.05). There were 42.86% (9/21) mutations in exon 7 of p53 gene and no mutation was observed in exons 5, 6, 8 in HCCs and in paracancerous tissues. In cases of p53 mutations, there were 66.67% (6/9) of p53 overexpression and 11.11% (1/9) of overexpression of both p53 and MDM2. MDM2 overexpression also appeared in 25% (3/12) of cases without mutation. CONCLUSIONS: Mutation of p53 gene and functional inactivation of p53 resulting from MDM2 overexpression play an important role in carcinogenesis of HCC. It is possible that p53 mutations and MDM2 overexpression induced by other mechanisms are involved in carcinogenesis of HCC.

[Effects of Stat3 phosphorylation and expression of c-fos and c-jun proteins on hepatocarcinogenesis].

Expression of phosphorylated Stat3 (p-Stat3), c-fos and c-jun proteins was detected by immunohistochemical technique in 55 hepatocellular carcinomas (HCC) and their surrounding liver tissues. The results showed that the positive rates and signal intensity of p-Stat3, c-fos and c-jun protein in HCCs were significantly higher than those in pericarcinomatous tissues. The expressive intensity of c-fos and c-jun proteins was positively related to p-Stat3 expression in HCCs, whereas the positive correlation of expressive intensity was only observed between c-jun protein and p-Stat3 in pericarcinomatous tissues. The data suggest that the phosphorylation of Stat3 may be an early event in hepatocarcinogenesis; the overexpression of p-Stat3 protein, which activates c-fos and c-jun genes, may contribute to malignant transformation of hepatocytes; hepatocytes which expressed p-Stat3, c-fos or c-jun proteins may be potentially malignant pre-cancer cells in pericarcinomatous liver tissues.

[Effect of hepatitis C virus NS3 protein on expression of iNOS mRNA in hepatocarcinogenesis].

OBJECTIVE: To investigate hepatocarcinogenesis by detecting the effect of HCV NS3 protein on expression of inducible nitric oxide synthase (iNOS) mRNA in hepatocellular carcinoma (HCC) and pericarcinomatous liver tissues(PCLT). METHODS: The expression of HCV NS3 protein and iNOSmRNA was detected by immunohistochemical technique (SP method) and in situ hybridization in specimens of HCC and PCLT from 52 patients with HBV(-). The relationship between the positive rate and the signal strength was tested statistically. RESULTS: The positive rate of HCV NS3 protein in HCC was lower (46.2% vs. 67.3%), and signal strength was weaker than those in PCLT. The signal strength of HCV NS3 protein was correlated with the degree of carcinomatous cells differentiation in HCC (P < 0.01). The positive rates (80.8% and 90.4%) of iNOSmRNA in HCC and PCLT had no statistical difference. Signal intensity of iNOSmRNA in HCC was weaker than that in PCLT. The higher HCVNS3 protein expression level was, the stronger iNOSmRNA signals in PCLT (P < 0.01). In particular, the localization and patterns of HCV NS3 protein were much similar to those of iNOSmRNA in the PCLT of some patients. CONCLUSIONS: HCV NS3 protein may exert its hepatocarcinogenic effect in the early stage on host cells by increasing product of nitric oxide, which may bring about transformation of hepatocytes.

[Expressions of p-MAPK, cyclin D1, p53 protein and their relationship in osteosarcoma].

OBJECTIVE: To detect the expressions of p-MAPK, cyclin D1 and p53 protein, and investigate their relationship in osteosarcomas. METHODS: SP immunohistochemical technique was used to detect the expressions of of p-MAPK, cyclin D1 and p53 protein among the 58 osteosarcomas and 14 osteoid osteomas samples. RESULTS: The positive rates of p-MAPK, cyclin D1 and p53 protein were 77.6% (45/58), 60.3% (35/58) and 44.8% (26/58) respectively in 58 osteosarcomas, and 21.4% (3/14), 21.4% (3/14) and 7.1% (1/14) in 14 osteoid osteomas. Their positive rates and expressive intensities in osteosarcomas were higher than those in osteiod osteomas. Their positive correlation was significantly observed between p-MAPK and cyclin D1 proteins (P < 0.01), and wasn't observed between p-MAPK and p53 proteins (P < 0.05) in osteosarcomas. CONCLUSIONS: MAPK phosporylation may be one of causes for activation of cyclin D1, which play an important role in osteosarcogenesis. The mutation of p53 oncogene may not result in osteosarcogenesis by MAPK signaling pathway. Detecting p-MAPK by immunohistochemistry may be one of diagnostic indexes for osteosarcoma and benign osteoma.

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance.

AIM: To study the effect of phosphorylation of MAPK and Stat3 and the expression of c-fos and c-jun proteins on hepatocellular carcinogenesis and their clinical significance. METHODS: SP immunohistochemistry was used to detect the expression of p42/44(MAPK), p-Stat3, c-fos and c-jun proteins in 55 hepatocellular carcinomas (HCC) and their surrounding liver tissues. RESULTS: The positive rates and expression levels of p42/44(MAPK), p-Stat3, c-fos and c-jun proteins in HCCs were significantly higher than those in pericarcinomatous liver tissues (PCLT). A positive correlation was observed between the expression of p42/44(MAPK) and c-fos proteins,and between p-Stat3 and c-jun but there was no significant correlation between p42/44(MAPK) and p-Stat3 in HCCs and their surrounding liver tissues. CONCLUSION: The abnormalities of Ras/Raf/MAPK and JAKs/Stat3 cascade reaction may contribute to malignant transformation of hepatocytes. Hepatocytes which are positive for p42/44(MAPK), c-fos or c-jun proteins may be potential malignant pre-cancerous cells. Activation of MAPK and Stat3 proteins may be an early event in hepatocellular carcinogenesis.

[Relationship between vascular endothelial growth factor expression and microvessel density in hepatocellular carcinomas and their surrounding liver tissue].

The relationship between vascular endothelial growth factor (VEGF) expression and microvessel density was studied with immunohistochemical method in hepatocellular carcinoma (HCC) and pericarcinomatous liver tissue. The positive rate of VEGF in HCCs was significantly lower than in surrounding liver tissues (66.7% vs. 85.4%, P < 0.05). There was no significant difference between HCC and pericarcinomatous liver tissue on expressive intensity of VEGF. The positive signal of VEGF was mainly localized in cytoplasma of cancer cells, pericarcinomatous hepatocytes, and vascular endothelial cells. The microvessel density in HCC was higher than in pericarcinomatous liver tissue and closely correlated to differentiated degree of cancer cells (rs = 0.5870; rs = 0.8235). The poorer cancer cell differentiation, the higher microvessel density. The results suggest that VEGF may not be the sole factor that stimulates angiogenesis in HCC genesis and development. To detect microvessel density in judging prognosis and biological behavior of HCC is more important than that of VEGF.

[The expression of p42/p44MAPK and cyclin D1 protein and their relationship in Hodgkin's disease].

The expressions of p42/44 mitogen-activated protein kinase(p42/p44MAPK) and cyclin D1 protein were detected by SP immunohistochemical technique in 40 cases of Hodgkin's disease(HD) and 13 cases of reactive hyperplasia(RH). The results showed that p42/p44MAPK and cyclin D1 protein were both overexpressed in RS/H cells of HD(positive rates were 72.5% and 70% respectively), and their positive rates were positively correlated(P < 0.01). The data suggest that MAPK cascades signal transduction may play an important role in genesis of HD. MAPK may induce overexpression of cyclin D1 protein and result in persistent proliferation and progression of RS/H cells.

[An improved deresining method for semithin sections embedded-epoxy resin].

Deresining must be performed for HE, specific histochemical staining and immunohistochemical detection on semithin-sections embedded-epoxy resin. The deresining process in general methods had to take 24 hours, and the semithin-sections were easily separated from slides. An improved method was introduced in this paper, by which deresining time was only 5 minutes, and it could also avoid the separation of semithin-sections from the slides. The staining result was very good, and the resolving power of images was high.