Objective To explore the anti-hepatic-fibrosis mechanism of Baogan Ning(BN).Methods Fifty Wistar rats were randomized into 5 groups: normal group,model group,high-and low-dosage BN groups(35.4 and 17.7 g/kg respectively),and colchicine group(0.11mg/kg).Rat models of hepatic fibrosis were induced by multiplex factors.Hepatic protein was extracted,and the expression of hepatic leptic receptors of OB-Rb,JAK2 and STAT3 was detected with Western Blot method.Results The protein expression was not obvious in the normal group,and the color of protein band in the medication groups was light but dark in the model group.Compared with the normal group,the expression of OB-Rb,JAK2 and STAT3 increased in the model group.However,the expression of OB-Rb,JAK2 and STAT3 decreased in the medication groups as compared with the model group.And the expression of OB-Rb,JAK2 and STAT3 decreased in the BN groups as compared with the colchicines group.Conclusion The anti-hepatic-fibrosis mechanism of Baogan Ning is probably related with the inhibition of OB-Rb expression,thus inhibit the JAK2/STAT3 message pathway.

Objective:To discuss the mechanism of Baoganning anti-liver fibrosis.Methods:liver fibrosis rat models were established by complex factors.Automatic biochemical analyzer was used to detect TC、TG and RIA isotope analysis was used to detect serum leptin.To observe the influence of Baoganning on TC、TG and leptin level of liver fibrosis model rats. Results:Compared with the normal group,levels of serum TC,TG decreased apparently in model and every drug groups(P0.05).Conclusion:Baoganning can effectively prevent and treat liver fibrosis in rats,which may be closely related to the improvement of liver function,adjustment of lipid and reduction of serum leptin.

Hepatocellular carcinoma(HCC)is one of the most common malignancies worldwide.Due to the difficulty of diagnosis in the early stage of HCC, most HCCs are diagnosed in intermediate-advanced stage.Moreover, the high invasion, metastasis and recurrence rate of HCC result in the high mortality of HCC.MicroRNAs(miRNAs) are a class of highly conserved, endogenous, small, non-coding ,single stranded RNA with the length of 22 nucleotides.There are plentiful of miRNAs in liver.MiRNAs not only can regulate the growth and development of liver, but also are closely related to the formation of HCC.In the process of HCC formation, miRNAs could function as oncogenes or tumor suppressor genes to regulate multiple biological processes related to HCC, including cell differentiation,proliferation,tumorigenesis,angiogenesis,invasion,and metastasis.With the intensive study of molecular mechanisms of miRNAs in the process of HCC formation, increasingly studies have revealed that miRNAs could become sensitive biomarkers and effective therapeutic targets for HCC.

AIM To study the effects of Biejiajian Pills (Colla Carapacis Trionycis,Asini Corii Colla,Nidus Vespae,etc.) on NF-κB,p65,p50 and IκB in NF-κB signaling pathway and target gene expression in HSC-T6 cells of rats.METHODS HSC-T6 cells were cultured with Biejiajian Pills drug serum for 24 hours,the expressions of p65,p50,VEGF and TIMP-1 mRNA were determined by qPCR;the expression of p65 was measured by immunofluorescence;the expressions of IκBα,IκBβ and α-SMA were determined by Western blot.RESULTS The Biejiajian Pills middle-,high-dose and positive control groups showed significantly lower expressions of p65,VEGF and TIMP-1 mRNA as compared with the blank control group and negative control group,the expressions of p50 mRNA among various groups showed no significant differences.But immunofluorescence showed that the expression of p65 in cytoplasm was decreased.Meanwhile,Biejiajian Pills showed significantly higher IκBα protein expression and obvious down-regulation of α-SMA expression in a dose-dependent manner,but had no significant influence on the expression of IκBβ.CONCLUSION Biejiajian Pills' therapeutic effects on hepatic fibrosis may be related to influencing NF-κB signaling pathway and inhibiting the expression of down-stream target gene.

OBJECTIVETo investigate the effect of Biejiajian Pills on Wnt signal pathway and its inhibitory gene (DKK-1 and FrpHe) expressions and explore the mechanism underlying the action of Biejiajian Pills to suppress the invasiveness of hepatocellular carcinoma.

METHODSTwenty-four Wistar rats were randomized equally into 3 groups for gavage of normal saline and Biejiajian Pills at 20- and 10-fold clinical doses for 3 days. Blood samples were then collected from the rats, and the serum was separated and added in HepG2 cell cultures. After 48 h of culture, the cells were collected to determine the cellular content of β-catenin protein using flow cytometry and detect DKK-1 and FrpHe mRNA expressions using qRT-PCR.

RESULTSHepG2 cells cultured in the presence of sera from rats fed with Biejiajian Pills showed significantly lowered β-catenin protein expression and obvious down-regulation of DKK-1 mRNA expression, and the effect was correlated with the doses of the drug administered. The expression of FrpHe mRNA showed no significant differences between the 3 groups.

CONCLUSIONSBiejiajian Pills can effectively inhibit the invasiveness and migration of hepatocellular carcinoma cells, which is closely related to decreased expressions of β-catenin and DKK-1 to cause block of the Wnt/β-catenin signal pathway.

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Male ; Proto-Oncogene Proteins ; metabolism ; Rats ; Rats, Wistar ; Wnt Proteins ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

OBJECTIVETo study the effect of Biejiajian Pills on Wnt signal pathway and the mechanisms underlying its action to suppress the invasiveness of hepatocellular carcinoma.

METHODSHepG2 cells cultured in the serum of rats fed with Biejiajian Pills for 48 h were examined for β-catenin expression using immunofluorescence, β-catenin/TCF4 complex activity with luciferase, and expressions of the downstream proteins cyclin D1 and MMP-2 using qRT-PCR.

RESULTSBiejiajian Pills-treated sera significantly reduced the expressions of cytoplasmic and nuclear β-catenin protein, cyclin D1 and MMP-2 proteins and lowered the activities of β-catenin/TCF4 complex.

CONCLUSIONBiejiajian Pills may serve as a potential anti-tumor agent, whose effect might be mediated by inhibiting the Wnt/β-catenin pathway.

Animals ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; Cyclin D1 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Rats ; Transcription Factor 4 ; Transcription Factors ; metabolism ; Wnt Proteins ; Wnt Signaling Pathway ; beta Catenin ; metabolism

OBJECTIVETo investigate the effect of Biejiajian Pills on the expressions of the signal molecules and target genes of Wnt signal pathway in HepG2 cells and explore the mechanisms by which Biejiajian pills suppress the invasiveness of hepatocellular carcinoma.

METHODSHepG2 cells were cultured for 48 h in the presence of serum collected from rats fed with Biejiajian Pills. The expressions of β-catenin, GSK-3β and P-GSK-3β in the cultured cells were assessed by Western blotting and the expressions of CD44v6 and VEGF were detected using immunohistochemistry.

RESULTSHepG2 cells cultured with the serum of rats fed with Biejiajian Pills showed lowered expressions of β-catenin protein both in the cytoplasm and the nuclei with also inhibition of phosphorylation of GSK-3β and reduced expression of CD44v6 and VEGF.

CONCLUSIONBiejiajian Pills can significantly reduce the expression of β-catenin by decreasing the phosphorylation of GSK-3β and blocking the Wnt/β-catenin signaling pathway to cause down-regulation of the target genes CD44v6 and VEGF, which may be one of the molecular mechanisms by which Biejiajian Pills suppress the proliferation and invasiveness of hepatocellular carcinoma.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Hep G2 Cells ; drug effects ; Humans ; Hyaluronan Receptors ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Phosphorylation ; Rats ; Vascular Endothelial Growth Factor A ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

Objective:To explore the clinical effects of unilateral secondary puncture percutaneous vertebroplasty (PVP) in the treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fractures (ASOTLF).Methods:A retrospective case-control study was conducted to analyze the clinical data of 193 patients with type ⅡA ASOTLF who had been admitted to Department of Spine Surgery, Honghui Hospital from February 2016 to October 2018. They were 71 males and 122 females, aged from 65 to 90 years [average, (73.9±4.3) years]. The segments injured were T10 in 21 cases, T11 in 27 cases, T12 in 44 cases, L1 in 48 cases, L2 in 29 cases, L3 in 14 cases, and L4 in 10 cases. Of them, 85 received unilateral secondary puncture PVP (observation group) and 108 did not (control group). The clinical effects were evaluated by comparing between the 2 groups the operation time, bone cement injection volume, intraoperative blood loss, hospital stay, and visual analogue scale (VAS) for back pain, spinal Oswestry disability index (ODI), anterior height of the injured vertebral body (AH) and kyphosis angle (KA) of the injured vertebra before operation, at 3 days after operation and the last follow-up. The bone cement leakage and fracture of adjacent vertebral body were observed.Results:All patients were followed up for 12 to 24 months (average, 15.8 months). There was no significant difference in the preoperative general data between the 2 groups, showing they were comparable ( P>0.05). The operation time and bone cement injection volume [(36.2±1.4) min and (5.5±0.7) mL] in the observation group were significantly longer or more than those in the control group [(32.3±1.7) min and (4.0±0.7) mL] ( P<0.05). There was no significant difference in the hospital stay or intraoperative blood loss between the 2 groups ( P>0.05). The VAS, ODI, AH and KA at 3 days after operation and the last follow-up were significantly improved compared with those before operation in both groups ( P<0.05). There was no significant difference in VAS, ODI, AH or KA between the 2 groups before operation or at 3 days after operation ( P>0.05). However, the VAS, ODI, AH and KA at the last follow-up in the observation group [(2.2±0.8) points, 19.2%±5.8%, (2.90±0.21) cm, and 12.2°±1.5°] were better than those in the control group [(3.1±0.9) points, 22.8%±5.3%, (2.41±0.15) cm, and 13.3°±1.2°]. There was no significant difference between the 2 groups in the incidence of postoperative bone cement leakage or that of adjacent vertebral fracture ( P>0.05). Conclusions:In the treatment of type ⅡA ASOTLF, unilateral secondary puncture PVP can result in satisfactory clinical effects, because it effectively promotes dispersion of bone cement and prevents re-collapse of the vertebra operated but does not increase the risks of bone cement leakage and adjacent vertebral fracture.

ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages. MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl₂）. The cells were randomly divided into blank group， CoCl₂ hypoxia model group （200 mmol·L^-1）， Biejiajian Wan low-dose group （200 mmol·L^-1+0.55 g·kg^-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L^-1+1.1 g·kg^-1 Biejiajian Wan）， and high-dose group （200 mmol·L^-1+2.2 g·kg^-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L^-1+0.56 g·kg^-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence. ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl₂ stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited. ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.

OBJECTIVE: To study the inhibitory effect of pills (BJJ) agaisnt diethylnitrosamine (DEN)-induced hepatocarcinogenesis and explore the relation between this effect and the inflammasome signaling pathway. METHODS: Sixty-five male SD rats were randomly divided into control group, DEN model group, and 3 BJJ treatment groups at low, medium and high dose (with daily dose of 0.55, 1.1 and 2.2 g/kg, respectively, for 12 consecutive weeks starting from the 5th week after modeling). The pathological changes of the liver tissue were observed with HE and Masson staining, and serum levels of alanine transaminase (ALT), glutamic oxaloacetic transaminase (AST), alkaline phosphatase (ALP) and total bilirubin (TBIL) of the rats were detected using ELISA. Oxidation stress in the liver tissue was assessed with ELISA, and Western blotting and ELISA were used to detect the molecular expressions of inflammasome-related pathway. RESULTS: BJJ significantly inhibited tumor growth in the liver of the rats. HE and Masson staining showed that BJJ treatment obviously ameliorated liver fibrosis and reduced cancer cell and inflammatory cell infiltration in the liver. BJJ significantly reduced elevations of serum ALT, AST, ALP and TBIL levels, increased the contents of superoxide dismutase, catalase and glutathione peroxidase in the liver and suppressed malondialdehyde in Den-treated rats. BJJ also dose-dependently decreased the expressions of NLRP3, apoptosis-associated speck-like protein (ASC), caspase-1, pro-IL-1β, pro-IL-18, IL-1β and IL-18 in the liver of Den-treated rats. CONCLUSIONS BJJ treatment can dose-dependently inhibit DEN-induced hepatocarcinogenesis by enhancing antioxidant capacity and down-regulating inflammatory-related pathways in rats.
Animals ; Aspartate Aminotransferases ; Diethylnitrosamine ; Liver ; Liver Neoplasms ; Male ; Rats ; Rats, Sprague-Dawley