Mechanism of Glycyrrhizae Radix et Rhizoma Alleviating Tripterygium wilfordii Polyglycoside Tablets-induced Liver Injury / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of cytochrome P4502D6 （CYP2D6） and cytochrome P4503A4 （CYP3A4）， key enzymes of drug metabolism in liver， on acute liver injury in water extract of Glycyrrhizae Radix et Rhizoma （WEOGRR）. MethodHealthy male Kunming mice were divided into normal group， model group， WEOGRR low-， medium- and high-dose groups （5， 10， 15 g·kg-1·d-1） and positive drug group （diammonium glycyrrhizinate， 75 mg·kg-1·d-1）， with 10 in each group. One week after preventive administration， acute liver injury model was induced by single intragastric administration of 270 mg·kg-1 Tripterygium Glycosides tablets， and samples were collected after 18 h. The pathological changes of liver were observed by hematoxylin-eosin （HE） staining. Serum liver function indexes including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyl transpeptadase （γ-GT）， alkaline phosphatase （ALP）， and total bilirubin （TBIL） as well as the levels of oxidative stress indexes including malondialdehyde （MDA） and superoxide dismutase （SOD） in hepatocytes were determined by biochemical method. Real-time polymerase chain reaction （Real-time PCR） and Western blot were performed to detect the mRNA and protein expression levels of CYP2D6 and CYP3A4， respectively. ResultCompared with normal group， model group had significant hepatocyte swelling and inflammatory cell infiltration （P<0.01）， increased AST， ALT， γ-GT， ALP and TBIL （P<0.05）， elevated MDA and decreased SOD （P<0.01） as well as down-regulated mRNA and protein expression levels of CYP2D6 and CYP3A4 （P<0.05）. Compared with the model group， the normal group had intact liver structure without obvious abnormality， and the WEOGRR groups and positive drug group presented alleviated hepatocyte swelling and inflammatory cell infiltration （P<0.01）， reduced AST， ALT， γ-GT， ALP and TBIL （P<0.01）， lowered MDA and increased SOD （P<0.01） as well as up-regulated expression levels of CYP2D6 and CYP3A4 （P<0.01）. ConclusionThe protective effect of WEOGRR on acute liver injury induced by Tripterygium glycosides tablets may be related to reducing the contents of AST， ALT， γ-GT， ALP and TBIL in serum， inhibiting MDA and increasing the activity of SOD in liver cells， and enhancing the activities of CYP2D6 and CYP3A4， thus accelerating the metabolism of toxic substances.

The osteogenesis ability of human umbilical cord Wharton's Jelly-derived mesenchymal stem cells and periodontal mesenchymal stem cells / 实用口腔医学杂志

Objective:To compare the osteogenesis ability between human umbilical cord Wharton's Jelly-derived mesenchymal stem cells(hUCWJMSCs) and human periodontal ligament mesenchymal stem cells (hPDLSCs) in vitro.Methods:hUCWJMSCs and hPDLSCs were in vitro cultured.The cell proliferation capacity was examined by MTT assay.After osteogenesis induction culture,ALP activity of the cells was determined,minerialization was observed by alizarin red staining,OPN and Runx2 mRNA expression was analyzed by Real-time PCR.Results:hUCWJMSCs grew faster than hPDLSCs.After osteogenic differentiation induction,hPDLSCs group showed higher ALP level,more mineralized nodule formation and higher Runx2 expression compared with hUCWJMSCs group (P ＜ 0.05);while the OPN expressed higher in hUCWJMSCs than in hPDLSCs (P ＜ 0.05).Conclusion:hUCWJMSCs and hPDLSCs have osteogenesis differentiation potential,hPDLSCs are more osteogenetic.

Explicit determination of the whole group of cervical cancer and genetic susceptibility studies / 重庆医学

Objective To investigate the expression levels of MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene in cervical cancer .Methods In this study ,128 patients with cervical cancer treated in our hospital from March 2010 to May 2014 were selected as the observation group ,and 100 cases of healthy people were selected as control group .MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene expression levels in cancer tissues were tested .(1) HIF‐1α:HIF‐1α kit was used to detect HIF‐1αmonoclonal antibodies ,the kit was prepared and stained according to the requirements ,and the positive cell rate greater than 10%under the microscope were positive .(2) MiR‐96 ,Twist and Slug:total RNA was extracted according to the instructions ,the RNA was reverse transcribed ,the relative expression values MiR‐96 ,Twist and Slug were detected by quantitative PCR .Results (1) HIF‐1αwas not expressed in normal tissues .And in tumor tissues ,the positive expression rate was much higher than that of normal tissue ,there was a significant difference(P<0 .05);(2)the relative expression values of MiR‐96 in tumor tissue were much greater than that of normal tissue (P<0 .05);(3) the relative expression rate of Twist and Slug gene in the tumor tissue were also much higher than that of normal organizations (P<0 .05) .Conclusion Compared with normal tissues ,MiR‐96 ,HIF‐1α,Twist and Slug gene expression in tumor tissues are significantly greater .

Using FRET to Study The Interaction Domain of TLR4 Binding to MD-2 in Living Cells / 生物化学与生物物理进展

TLR4-MD-2 complex plays a key role in LPS recognition and its signal transduction. These steps are the vital elements of the host's defensive reaction. Studying the functional domain of TLR4 and MD-2 is very important to further understand the mechanism of LPS signal transduction. It was studied the interaction domain of TLR4 and MD-2 in living cells based on gene mutation, gene transfection and fluorescence resonance energy tramsfer(FRET) which is considered as one of the best methods used for intracellular protein-protein interaction study. CY-15P which was fused by CFP and YFP through 15 neutral amino acids was used as positive control, while co-expressed CFP and YFP proteins were used as negative control. The results showed that the ability of TLR4 binding to MD-2 decreased dramatically after the deletion of Glu~(24) ～ Met~(41) in N terminal of TLR4. Aggregation of TLR4 to LPS stimulation was observed, however, TLR4 without the Glu~(24)～ Met~(41) mutation did not aggregate. All these results indicated that TLR4 Glu~(24)～ Met~(41) might be the interaction domain of TLR4 binding to MD-2 and participate in the aggregation effect of TLR4 upon LPS stimulation.

The Role of MD-2 in The Process of Endotoxin Recognition and Signal Transduction / 生物化学与生物物理进展

Lipopolysaccharide(LPS) can induce cell inflammation through interacting with TLR4. Recent studies have revealed that MD-2 participate in the process of LPS induced signal transduction pathway by forming a complex with TLR4. After binding to the MD-2 of the TLR4/MD-2 complex, LPS can induce TLR4- oligomerization and activate the downstream signal pathway. After being synthesized, most MD-2 can bind to TLR4 at the endoplasmic reticulum /Golgi apparatus and expresse as TLR4/MD-2 complex at the cellular surface. Therefore MD-2 not only can regulate the distribution of TLR4 in the cytoplasm, but also help TLR4 to recognize LPS. Another part of MD-2 can be released into plasma as soluble MD-2(sMD-2). With the help of CD14, sMD-2 would interact with LPS in the plasma to constitute LPS-sMD-2 complex, helping cell who express only TLR4, to recognize LPS, however excessive expressed sMD-2 would repress the LPS signal transduction pathway. In conclusion, MD-2 plays a crucially modulating role in the process of TLR4 mediated endotoxin recognition and signal transduction.