Constitutive androstane receptor induced-hepatomegaly and liver regeneration is partially

The constitutive androstane receptor (CAR, NR3I1) belongs to nuclear receptor superfamily. It was reported that CAR agonist TCPOBOP induces hepatomegaly but the underlying mechanism remains largely unknown. Yes-associated protein (YAP) is a potent regulator of organ size. The aim of this study is to explore the role of YAP in CAR activation-induced hepatomegaly and liver regeneration. TCPOBOP-induced CAR activation on hepatomegaly and liver regeneration was evaluated in wild-type (WT) mice, liver-specific YAP-deficient mice, and partial hepatectomy (PHx) mice. The results demonstrate that TCPOBOP can increase the liver-to-body weight ratio in wild-type mice and PHx mice. Hepatocytes enlargement around central vein (CV) area was observed, meanwhile hepatocytes proliferation was promoted as evidenced by the increased number of KI67

Facile synthesis of insulin fusion derivatives through sortase A ligation

Insulin derivatives such as insulin detemir and insulin degludec are U.S. Food and Drug Administration (FDA)-approved long-acting insulin currently used by millions of people with diabetes. These derivatives are modified in C-terminal B29 lysine to retain insulin bioactivity. New and efficient methods for facile synthesis of insulin derivatives may lead to new discovery of therapeutic insulin. Herein, we report a new method using sortase A (SrtA)-mediated ligation for the synthesis of insulin derivatives with high efficiency and functional group tolerance in the C-terminal B chain. This new insulin molecule (Ins-SA) with an SrtA-recognizing motif can be conjugated to diverse groups with N-terminal oligoglycines to generate new insulin derivatives. We further demonstrated that a new insulin derivative synthesized by this SrtA-mediated ligation shows strong cellular and

Characterization of organic anion transporting polypeptide 1b2 knockout rats generated by CRISPR/Cas9: a novel model for drug transport and hyperbilirubinemia disease

Organic anion transporting polypeptide 1B1 and 1B3 (OATP1B1/3) as important uptake transporters play a fundamental role in the transportation of exogenous drugs and endogenous substances into cells. Rat OATP1B2, encoded by the gene, is homologous to human OATP1B1/3. Although OATP1B1/3 is very important, few animal models can be used to study its properties. In this report, we successfully constructed the S knockout (KO) rat model using the CRISPR/Cas9 technology for the first time. The novel rat model showed the absence of OATP1B2 protein expression, with no off-target effects as well as compensatory regulation of other transporters. Further pharmacokinetic study of pitavastatin, a typical substrate of OATP1B2, confirmed the OATP1B2 function was absent. Since bilirubin and bile acids are the substrates of OATP1B2, the contents of total bilirubin, direct bilirubin, indirect bilirubin, and total bile acids in serum are significantly higher in KO rats than the data of wild-type rats. These results are consistent with the symptoms caused by the absence of OATP1B1/3 in Rotor syndrome. Therefore, this rat model is not only a powerful tool for the study of OATP1B2-mediated drug transportation, but also a good disease model to study hyperbilirubinemia-related diseases.

Effect and mechanism of total flavonoids of Lichi Semen on CCl_4-induced liver fibrosis in rats, and prediction of Q-marker / 中国中药杂志

This paper was to investigate the effect of total flavonoids of Lichi Semen(TFL) on carbon tetrachloride(CCl_4)-induced liver fibrosis in rats, analyze and predict its mechanism of action and potential quality markers(Q-marker). Firstly, male SD rats were taken and injected subcutaneously with a 40% CCl_4-vegetable oil solution twice a week for 8 consecutive weeks to establish a rat model of liver fibrosis. The rats with liver fibrosis were randomly divided into model group, silybin group(43.19 mg·kg~(-1)), Fuzheng Huayu Capsules group(462.75 mg·kg~(-1)), and TFL groups(100 mg·kg~(-1) and 25 mg·kg~(-1)), with normal rats as a blank group, 10 rats in each group. Except for the blank group, the rats in the other groups were subcutaneously injected with 40% CCl_4-vegetable oil solution of a maintenance dose, once a week. The rats in various treatment groups received corresponding doses of drugs, while the rats in the blank group and model group received the same volume of normal saline once a day for 4 weeks. At the end of the experiment, blood was collected from the abdominal aorta and the liver tissues were collected. The levels of total bilirubin(TBiL), direct bilirubin(DBiL), indirect bilirubin(IBiL), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) in serum were detected by using an automatic biochemical detector. Masson staining was used to observe the histopathological changes of rat liver. Then, the chemical compositions of TFL were collected, and the action targets of these chemical compositions were predicted through SWISS database and reverse molecular docking server(DRAR-CPI). After screening of disease targets of liver fibrosis by Gene Cards database, the protein-protein interaction was analyzed with use of STRING database, and GO(gene ontology) analysis and KEGG(Kyoto encyclopedia of genes and genomes) enrich analysis were also carried out. Moreover, an iTRAQ proteomics technology was used to determine protein expression in liver tissues of rats in TFL, model and blank groups to verify the targets. Furthermore, Cytoscape software was used to establish and visualize the network of chemical components, targets and pathways, and predict the potential Q-marker of TFL. The results showed that the levels of TBiL, DBiL, IBiL, ALT, and AST in the model group were significantly higher than those in the blank normal group(P<0.05), and the above levels in the treatment groups were lower than those in the model group, but with no significant differences. Masson staining showed that the liver damage and the degree of fibrosis were severe in the model group, and were relieved to different degrees in the treatment groups. Then, 74 chemical components were screened, which could act on 865 targets such as EGFR and SRC, participating in the regulation of cancer pathways, PI3 K-Akt signaling pathway, HIF-1 signaling pathway and other signaling pathways closely related to liver fibrosis. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin showed the highest correlation with liver fibrosis-related targets and pathways. Proteomics results showed that a total of 18 proteins among the 45 proteins predicted by internet pharmacology were identified, among which 6 proteins were significantly expressed, including 5 up-regulated proteins and 1 down-regulated protein. The protein expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1 was significantly returned to a normal state in the TFL treatment groups. In conclusion, TFL may demonstrate the anti-hepatic fibrosis and potential hepatoprotective effects by regulating the expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1, which may be associated with the regulation of multiple signaling pathways related to liver fibrosis such as PI3 K-Akt pathway. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin could be regarded as potential Q-markers of TFL for quality control.

Effect of Fugan Huaxian Decoction on MAPK signaling pathway in model rats with hepatic fibrosis / 中草药

Objective: To research the effect of Fugan Huaxian Decoction on MAPK signaling pathway in rats with hepatic fibrosis (HF) and explore the mechanism of anti-HF. Methods: A total of 120 SD rats were randomly divided into control group, model group, colchicine group and Fugan Huaxian Decoction group (high, medium and low dose groups), with 20 rats in each group. The rats in the normal group were SD rats, and the rats model in the remaining groups were established into HF rats with syndrome of qi deficiency, poison weakness and blood stasis on the basis of the HF model induced by carbon tetrachloride (CCl4). Moreover, the rats were also received tail clamping, forced swimming, abnormal of starvation and full as well as rhubarb gavage. Liver pathology was performed on all rats after six weeks of modeling. After the validation model was successful, each group was given different doses of gavage, colchicine group (2 mg/kg), high, medium and low dose groups of Fugan Huaxian Decoction were intra-gastrically administered (67.08, 33.54, and 16.77 g/kg), normal group and model group were given pure water 2 mL/d for three weeks continuously. On the second day of last gavage, the serum levels of ALT, AST, ALB, LN, HyP and PIIINP were detected in each group. HE staining and Masson staining were performed on liver tissues. The expression of ERK1/2, JNK1/2 and P38-related protein and their phosphorylated protein in liver MAPK signaling pathway were detected by Western blotting. The results of quantitative analysis were expressed as p-ERK/ERK, p-JNK/JNK, p-p38/p38 ratio. Results: In model group, hepatic tissue cells were severely infiltrated and fibrotic, while, the degree of liver injury and fibrosis were significantly reduced in other groups. Compared with normal group, the serum levels of ALT, AST, Hyp, LN and PIIINP in model group were increased significantly, ALB was decreased significantly (P < 0.01), and the protein expression of p-JNK, p-ERK and p-p38 was increased significantly (P < 0.01). Compared with model group, Fugan Huaxian Decoction decreased serum ALT, AST, Hyp, LN and PIIINP levels of rats, increased ALB content and down-regulated the expression of phosphorylated protein of p38, ERK1/2 and JNK1/2, and it showed in a dose-dependent manner, the high dose group worked the best. Conclusion: The model of qi deficiency, poison weakness and blood stasis combined with HF model rats was successfully established. Fugan Huaxian Decoction not only protects hepatocytes, alleviates liver injury and inhibits HF, but also regulates the protein expressions of p-JNK, p-ERK, p-p38, as well as inhibits MAPK signaling pathway activation, which may be one of the mechanisms of its anti-HF function.

Mouse models for hepatitis B virus research / 한국실험동물학회지

Hepatitis B virus (HBV) infection remains a major global health problem; indeed, there are 250 million carriers worldwide. The host range of HBV is narrow; therefore, few primates are susceptible to HBV infection. However, ethical constraints, high cost, and large size limit the use of primates as suitable animal models. Thus, in vivo testing of therapies that target HBV has been hampered by the lack of an appropriate in vivo research model. To address this, mouse model systems of HBV are being developed and several are used for studying HBV in vivo. In this review, we summarize the currently available mouse models, including HBV transgenic mice, hydrodynamic injection-mediated HBV replicon delivery systems, adeno-associated virus-mediated HBV replicon delivery systems, and human liver chimeric mouse models. These developed (or being developed) mouse model systems are promising and should be useful tools for studying HBV.

Relationship between ischemia-modified albumin and coronary stenosis / 西安交通大学学报(医学版)

Objective To investigate the relationship between ischemia-modified albumin (IMA) and coronary artery stenosis in patients without myocardial infarction.Methods For this study we consecutively enrolled 345 patients who received coronary angiography (CAG).According to the results,the subjects were divided into coronary stenosis group (232 cases)and control group (113 cases)to investigate the the relationship of IMA and IMA/albumin (IMAr)with coronary stenosis.Results ① The levels of IMA and IMAr in coronary stenosis group were higher than those in control group (P＜0.001).② The IMA and IMAr were higher in single-branch and multi-branch lesion groups than in control group (P＜0.05),whereas there was no significant difference between single-branch lesion group and multi-branch lesion group (P＞0.05).③ In receiver operating characteristics curve analysis,the sensitivity of IMA and IMAr was 64.4% and 78.0%,respectively (AUC:0.653,0.705,P＜0.001)in predicting coronary stenosis.④ Multivariate logistic regression analysis showed that IMAr was an independent risk factor for coronary stenosis in patients without myocardial infarction (OR=73.05,P＜0.001).Conclusion IMA and IMAr are closely correlated with coronary stenosis and have a value in predicting coronary artery stenosis in patients without myocardial infarction.

Correlation between preoperative CRP/Alb ratio and lymph node metastasis in pa-tients with gastric cancer / 中国肿瘤临床

Objective:To investigate the relationship between Creactive protein(CRP)/albumin ratio (CAR) and lymph node metastasis of gastric cancer. Methods:A total of 96 cases of gastric cancer were included in the study. The clinical pathological stage and lymph node detection in patients with gastric cancer were analyzed, with the preoperative CAR as the dependent variable. Results:1) The pa-tients with preoperative CAR>0.04 has higher transfer rate and metastasis than the patients with preoperative CAR≤0.04, and the dif-ference was significant (P0.04 was significantly higher than that of the patients with preoperative CAR≤0.04, and the difference was significant (P<0.05). 2) The mean CAR of patients with Borrmann typesⅠ,Ⅱ,Ⅲ, andⅣgradually increased;with the progression of the pathological stage of gastric cancer, the mean value of CAR in-creased. 3) During the operation, the total number of lymph node dissections was high and the mean value of CAR before the opera-tion was high. Conclusion:The correlation between preoperative CAR and lymph node metastasis in patients with gastric cancer may reflect the degree of lymph node metastasis to a certain extent.

Establishment of an Alb-cre/DTR mouse model of inducible liver injury / 中国实验动物学报

Objective To analyze the Alb-cre/DTR mouse phenotype, and establish a model of induced liver damage to serve basic researches of liver diseases.Methods The introduced Alb-cre and DTR mice were crossed to obtain Alb-cre/DTR mice and the genomic DNAs were extracted from the tail tissue of the mice for genotying by PCR.Diphtheria toxin was intraperitoneally(i.p.)injected into the Alb-cre/DTR mice, then the body weights were monitored and the sera were collected for the detection of serum ALT and AST levels.Results By crossing Alb-cre and DTR mice we obtained the Alb-cre and DTR double transgenic mouse.The intraperitoneal injection of diphtheria toxin in a dose of 0.625 ng/g body weight significantly induced liver injury in these mice, as showed by the elevated levels of ALT and AST, the gross appearance of liver damage and the pathological changes such as necrosis in the liver tissue.Conclusions We have ob-tained a novel mouse strain of Alb-cre/DTR by crossing Alb-cre and DTR mice.Liver damages in those Alb-cre/DTR mice can be induced by injection of diphtheria toxin.This established mouse model of inducible liver damage is a useful platform for the studies of liver damage and recovery, as well as liver transplantation.

Analysis of lignan components in Schizandrae Fructus residue extracts and their protective effects on acute-hepatic damnification rat / 中草药

Objective: The lignan components in Schizandrae Fructus residue extracts were analized and evaluated. The protective effect of Schizandrae Fructus residue extracts on acute-hepatic damnification rat and the influence of that on intestinal flora were researched in the process of Shengmai Injection production, in order to provide the scientific basis for the development and utilization of Schizandrae Fructus extract. Methods: Schizandrae Fructus residue extract was extracted by the 60% ethanol. The model of acute-hepatic damnification rats induced by CCl4 was prepared, the protective effects of Schizandrae Fructus residue extracts on hepatic-damnification rats were evaluated through the data of ALT, AST, ALB, and TP; The function of the extracts were evaluated by histopathological observation of the liver and colon of rats; Effect of extracts on hepatic-damnification rat's intestinal flora was evaluated by the number and the number variation of the intestinal flora. Results: The results showed that the mass fractions of schisandrin A, schizandrin B, schizandrin C, gomisin A, gomisin B, and schisantherin A were 1.44, 3.42, 1.68, 4.22, 2.92, and 0.58 mg/g, respectively. Pharmacodynamic evaluation showed that Schizandrae Fructus extracts can significantly reduce the content of AST and ALT (P < 0.01) and increase the content of ALB and TP in a certain extent in the serum of acute-hepatic damnification rats (P < 0.05); Mucous damage score of liver and colon is significantly decreased; Compared to liver injury group, Schizandrae Fructus extracts of high dose group and low dose group can significantly promote the proliferation of Lactobacillus and Bifidobacterium (P < 0.05) but reduce the number of Enterococcus and Enterobacteria in the intestinal tract of rats (P < 0.05). Conclusion: Schizandrae Fructus residue extracts have the protective effects on the acute-hepatic damnification rats induced by CCl4, the effect of regulating intestinal flora and the proliferation of probiotics, the effect of protecting the constitution integrity of colon tissue, which provide a main basis for its further development and utilization.

Diagnostic value of HbA1 c combined with u -ALB in early type 2 diabetes / 中国基层医药

Objective To investigate the value of glycated hemoglobin(HbA1c)and urine trace albumin (u -ALB)content in early diagnosis of type 2 diabetic nephropathy.Methods 200 patients with type 2 diabetes were selected as diabetes group,and 30 cases of healthy people as control group.According to the content of HbA1c, the diabetes patients were re -divided into low group (HbA1c 10.1%).The levels of HbA1c and u -ALB were detected and the correlation between them was calculated.Results The values of HbA1c (8.85 ±1.22)% and u -ALB (88.3 ±12.4)mg/L in the diabetes group were significantly higher than those of the control group (t =10.88,54.25,all P <0.05);The level of HbA1c in the high,medium and low group[(11.02 ±1.37)%,(8.45 ±2.01)%,(6.88 ±1.23)%]were consistent with the levels of the respective levels of fasting glucose[(13.22 ±2.05)mmol/L,(9.25 ±1.28)mmol/L, (6.27 ±0.63)mmol/L].HbA1c and constituting the u -ALB levels were positively correlated in the high,medium and low group(r =0.452,0.512,0.452,all P <0.05).Conclusion The combined detection of HbA1c and u -ALB levels has important value for early diagnosis of type 2 diabetic nephropathy.

Kallistatin, a new and reliable biomarker for the diagnosis of liver cirrhosis

Kallistatin, which protects organs and cells against inflammation, fibrosis and oxidative stress, is mainly synthesized and secreted in liver. However, its relationship to human liver disease remains unclear. The purpose of this study was to explore the relationship between serum kallistatin and clinical evidence of both cirrhosis and hepatocellular carcinoma (HCC), and to determine if serum kallistatin levels could be used as a diagnostic indicator of hepatic health status, especially human liver cirrhosis (LC). Our cohort consisted of 115 patients with clinically proven liver fibrosis (LF), LC, or HCC by liver biopsies, and 31 healthy controls (CON). Serum kallistatin levels were quantified by ELISA. Results of the present study demonstrated that irrespective of the underlying etiology, serum kallistatin levels were significantly lower in the LF/LC group when compared with the CON group. A decrease in serum kallistatin levels appeared to reflect the extent of cirrhosis, with the lowest levels associated with higher grades of cirrhosis. Patients with LC had a noticeable correlation between serum kallistatin levels and other serum biochemical indicators. The area under the curve (AUC) for LC, viral liver cirrhosis (VLC) and alcoholic liver cirrhosis (ALC) was 0.845, 0.757 and 0.931, respectively. In conclusion, our findings demonstrated that kallistatin, a plasma protein produced by the liver, can be a useful and reliable diagnostic indicator of hepatic health status, especially for LC.

The establishment of the uPA inducible expression system / 中国比较医学杂志

Objective To establish uPA inducible expression system using recombinant retroviral system for the further construction of inducible uPA-SCID animal model .Methods The Inducible expression system need to construct two plasmids:pLNHXO1O2-Alb-GLUC-FMN2A -rtTA and pLNHXO5O6-TRE2-uPA-IRES-ZsGreen respectively. Both plasmids were based on retroviral vector pLNHX , Albumin promoter gene ( Alb) and rtTA gene or uPA gene and ZsGreen were obtained by PCR reaction and inserted into pLNHX .The Gaussia enzyme fluorescent element ( GLUC) was used to monitor rtTA expression in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA, and the ZsGreen for uPA expression monitoring in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen.The correct constructed plasmids were transfected into packaging cell line GP 2-293 to gain recombinant viral particles .NIH/3T3 cells were infected with these viral particles and selected with G 418.Gene expression in the surviving cells was confirmed by the PCR method .Results The recombinant retroviral vectors harbouring target genes were successfully cloned .The rtTA gene in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA was expressed, and uPA can be induced to express in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen by doxycycline (Dox) when the plasmid transfected into the HepG-Tet-on cell.The constructed recombinant two retroviral vectors were transfected into GP 2-293 packaging cells respectively to gain infectious viral particles .Then,NIH/3T3 cells were infected with these viral particles and single-cell clones which stably expressed the transgenes were successfully established .Conclusion This study primarily established uPA inducible expression system , it laid a foundation for the murine model of inducible liver damage , and provided a novel technical platform for further building the liver humanised murine models for viral hepatitis studying .

Influencing factors of whole blood colloid osmotic pressure in critically ill patients / 中南大学学报(医学版)

Objective: To analyze the influencing factors of whole blood colloid osmotic pressure (COP) and predict reference range of plasma protein for safe COP to guide clinical infusion of protein in critically ill patients. Methods: Physical data and blood gas analysis of 405 patients were collected. hTe patients were divided into 2 groups by COP: group A (COP≤18 mmHg) and group B (COP>18 mmHg). The serum proteins including total protein (TP), albumin (Alb), globumin (Glb) and ifbrinogen (FIB) were detected. Results: APACHE II of group B was signiifcantly lower than that of group A (P23.3 g/L,Glb 12.6-37.6 g/L and FIB 1.3-8.7 g/L; 5 reformed equations were made. Conclusion: The main influencing factors of COP include Alb, Glb and FIB. We can use the reference range of 4 types of protein level to guide the clinical management of protein agents, and reformed equations can be used to preliminarily forecast COP in critically ill patients.

Clinical Characteristics and ALB Gene Mutation Analysis of Korean Patients with Bisalbuminemia / 대한진단검사의학회지

BACKGROUND: Bisalbuminemia is a hereditary or an acquired condition characterized by the presence of 2 albumin variants with different mobilities on serum protein electrophoresis (SPE). The clinical significance of bisalbuminemia has not been clearly established. However, some regions of the albumin variant may affect the biochemical analysis of biomolecules such as steroid or thyroid hormones by altering their albumin-binding affinities. In this study, we analyzed the clinical manifestations, genetic variations, and the albumin-binding characteristics in Korean patients with bisalbuminemia. METHODS: We performed SPE for samples from 580 Korean subjects and identified bisalbuminemia on the basis of the results of SPE. The clinical and biochemical characteristics, ALB gene mutations, and the structures of the albumin variants of patients with bisalbuminemia were analyzed. RESULTS: SPE showed bisalbuminemia in 2 patients. One patient showed a genetic variation known as Nagasaki-1 (Asp293Gly) and the other showed a hitherto unreported missense mutation (c.593A>T; Lys198Ile). In both cases, the serum concentrations of the substances with binding affinity for albumin were not affected, and the mutation sites of the albumin were not located with the protein-binding loci. CONCLUSIONS: The 2 Korean patients with bisalbuminemia showed genetic variations, including a novel missense mutation. The ALB gene analysis with 3D modeling is useful for determining the nature of bisalbuminemia and for predicting the effects on the albumin-binding affinity of other biochemical compounds.

The Significance of Immunoglobulins and Albumin in the Cerebrospinal-fluid (CSF) on Differential Diagnosis of Meningitis / 中国医师杂志

61mg/L. Conclusion The destruction of blood-brain barrier in TM and PM was obviously stronger than that in VM. The contents of CSF-Ig and -Alb are helpful to diagnose and differntially diagnose meningitis.

Study on the Hemostatic Effect of Extracts from Herb Lamiophlomis rotata and Its Mechanism / 中药新药与临床药理

Objective To study the effect of Tibet medicine of Herba Lamiophlomis rotata on promoting hemostasis and blood coagulation and to explore its mechanism. Methods High dose(2 g/kg),middle dose(1 g/kg) and low dose (0.5 g/kg) of the aqueous extract from Herba Lamiophlomis rotata(HLRE)were given to mice by gastric gavage for 3 days,and then bleeding time (BT),clotting time(CT) and platelet count(PLC)were determined. Different doses of HLRE(3,1.5,0.75 g/kg)were administered by gastric gavage to rats for 14 days,and then blood samples were collected from common carotid artery for the determination of prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time(TT),fibrinogen(FIB),hepatic function and blood lipid indexes. Results Compared to the blank control group,the bleeding time and clotting time were obviously shortened in 2g/kg HLRE and 1g/kg HLRE groups and positive control group,but the difference of PLC was insignificant. PT and APTT values in all of the treatment groups did not differ from those in the blank control group. However, TT values were obviously shortened in 3 g/kg HLRE group,FIB and Albumen(Alb)values increased,the aminotransferase and blood lipid values tended to decrease. Conclusion The aqueous extract from Herba Lamiophlomis rotata has an effect on promoting hemostasis and blood coagulation ,and its mechanisms may be related to the increase of FIB and Alb contents and the shortening of TT value.

Effect of Repeated Weight Attached Swimming on Renal Ultrastrueture and Function in Rats / 中国运动医学杂志

The alterations of ultrastrueture in rat kidney were observed under transmission electron microscopy, results showed that both glomerular filtration membranes and renal tubules had changed immediately after repeated loaded swimming and there were consistency between the recovery of renal ultrastructures and that of urinary protein and excration rate.