Objective To study the properties of stress relaxation of the middle and lower cervical vertebrae, and to evaluate the effects of laminotomy and anterior diskectomy and spinal fusion (ASF) on the properties . Methods The dynamic responses of stress relaxation of six intact and post operative fresh human cadaveric cervical vertebrae were measured in vitro. Results Under the condition of the constant strain, the functional equations and the curves of stress relaxation of the six intact and post operative fresh human cadaveric cervical vertebrae were obtained. The Gex(t)(the ratio of the ensuing instant stress to the original stress) of the post operative ones was significantly bigger than that of the intact ones; The Gex(t) of the laminotomic ones was significantly bigger than that of the ones having undergone the anterior discectomy and spinal fusion(ASF). Conclusion Either in flexion or in extension, cervical vertebrae have the similar behaviors of stress relaxation. The laminotomy and anterior discectomy and spinal fusion (ASF) all reduce the stress relaxation effect of the cervical vertebrae, but ASF is more significant.

OBJECTIVE: To identify MARCO gene knockout mouse model by genotyping,sequencing and Western blotting.METHODS: A total of 16 base-knockout MARCO~(-/-)C57 BL/6 mice( 8 female and 8 male) were obtained by clustered regularly interspaced short palindromic repeats( CRISPR)/CRISPR associated protein 9( Cas9) technique with MARCO~(+/+)C57 BL/6 mice(8 female),and their offspring MARCO~(+/-)mice were obtained. Then MARCO~(+/-)mice were inter-crossed to get a sufficient number of MARCO~(-/-)homozygous mice. The genotypes of mice were identified by gene sequencing and the relative expression of MARCO protein was detected by Western blotting. RESULTS: After one year of breeding,a total of 5 generations were bred. There were 5 types of MARCO~(-/-)genotypes(-11,-25,-36,-46,-61 bp) stably inherited; MARCO~(-/-)∶ MARCO~(+/-)∶ MARCO~(+/+)= 1 ∶ 2 ∶ 1,which was consistent with Mendelian's law of heredity. Using MARCO(-11 bp) as an example,42 MARCO~(-/-)mice,92 MARCO~(+/-)mice and 48 MARCO~(+/+)mice were obtained from the 5 th generation( F5 generation); and there was no significant difference in body mass of the above 3 genotypes of F5 generation mice at the 4 th,the 6 th and the 8 th weeks after birth( P > 0. 05). The relative expression of MARCO protein in MARCO~(-/-)(-11 bp) and MARCO~(-/-)(-46 bp) mice was significantly down-regulated,compared with that of MARCO~(+/+),MARCO~(-/-)(-36 bp) and MARCO~(-/-)(-61 bp) mice(P < 0. 05). MARCO~(-/-)(-11 bp)and MARCO~(-/-)(-46 bp) mice were chosen as the MARCO gene knockout mice. CONCLUSION: MARCO gene knockout mice were successfully identified,which laid a foundation for further study on the role and regulatory mechanism of MARCO gene in silicotic fibrosis in mice.

【Objective】 To investigate the effect of HLA-G expressed in platelets on Tax protein of human T cell leukemia type 1 virus (HTLV-1). 【Methods】 Platelets were isolated from anticoagulant whole blood, and HLA-G molecule on platelet membrane was detected by flow cytometry. The content of secretory HLA-G before and after platelet lysis was detected by ELISA, HTLV-1 human lymphoma cells MT2 were cultured with platelet lysate (PL). The effect of HLA-G in platelets on the expression of HTLV-1 protein Tax was evaluated by Western blot (WB). 【Results】 Membrane type mHLA-G was highly expressed on the surface of platelet membrane. The expression of secretory sHLA-G (ng/mL) increased after platelet lysis (15.73±1.01) vs (6.65±0.47), the expression of sHLA-G increased with the increase of platelet concentration in a dose-dependent manner. Compared with fetal bovine serum, PL significantly promoted the high expression of HLA-G protein and HTLV-1 virus tax protein in MT2 cells, and the addition of anti-HLA-G antibody to PL could effectively inhibit the expression of Tax and HLA-G protein. 【Conclusion】 High expression of immune tolerance molecule HLA-G on platelets can induce high expression of HTLV-1 protein Tax in human lymphoma cell MT2, which contributes to viral infection.

Lonicerae Japonicae Flos is a commonly used traditional Chinese medicine with a long history. It has the functions of detoxification, heat dissipation and heat dissipation, with a high medicinal value. It is mainly distributed in Henan, Shandong, Guangdong and other places. Researches have shown that the chemical constituents of honeysuckle mainly include organic acids, flavonoids, iridoid glycosides, three terpenoid glycosides, three terpenoid glycosides and volatile oils. And the pharmacological effects of honeysuckle are mainly anti-inflammation and antipyretic, anti-tumor, anti-bacteria, anti-virus, anti-aging and anti-oxidation, lowering blood sugar, protecting liver, protecting lung, neuroprotective, enhancing immune function and anti-platelet aggregation. Because of its rich pharmacological effects and high medicinal value, it has been used in a variety of prescriptions, with wide clinical applications and its large social demands. In this paper, we have reviewed the literatures on the chemical constituents and pharmacological effects of honeysuckle in recent years, and learned that many scholars have studied it, isolated a variety of new chemical components from honeysuckle for the first time and studied the pharmacological effects of honeysuckle in detail. In this paper, we reviewed the research progress of chemical constituents and pharmacological effects of Lonicerae Japonicae Flos, providing references for more people to learn related knowledge of honeysuckle and a scientific basis for the development and utilization of Lonicerae Japonicae Flos.

OBJECTIVE: To investigate the effect and mechanism of glucosides of chaenomeles speciosa (GCS) on ischemia/reperfusion-induced brain injury in mouse model. METHODS: Fifty 8-week C57BL/C mice were randomly divided into five groups with 10 in each group:sham group, model group, GCS 30 mg/kg group, GCS 60 mg/kg group and GCS 90 mg/kg group, and the GCS was administrated by gavage (once a day) for 14 d. HE staining was performed to investigate the cell morphology; the Zea-Longa scores were measured for neurological activity; TUNEL staining was performed to investigate the cell apoptosis; ELISA was used to detected the oxidative stress and inflammation; Western Blot was performed to investigate the key pathway and neurological functional molecules. RESULTS: Compared with the sham group, the brain tissues in model group were seriously damaged, presenting severe cell apoptosis, oxidative stress and inflammation, associated with increased NF-κB P65 and TNF-α levels as well as decreased myelin associate glycoprotein (MAG) and oligodendrocyte-myelin glycoprotein (OMgp)levels (all <0.01). Compared with the model group, the brain tissues in GCS groups were ameliorated, and cell apoptosis, oxidative stress and inflammation were inhibited, associated with decreased NF-κB P65 and TNF-α levels as well as increased MAG and OMgp levels (all <0.01), which were more markedly in GCS 60 mg/kg group. CONCLUSIONS GCS can inhibit the NF-κB P65 and TNF-α, reduce the oxidative stress and inflammation, decrease the cell apoptosis in mouse ischemia/reperfusion-induced brain injury model, and 60 mg/kg GCS may be the optimal dose.
Animals ; Brain ; drug effects ; Brain Injuries ; drug therapy ; Gene Expression Regulation ; drug effects ; Glucosides ; pharmacology ; therapeutic use ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; genetics ; Oxidative Stress ; drug effects ; Plant Extracts ; pharmacology ; Random Allocation ; Rosaceae ; chemistry ; Tumor Necrosis Factor-alpha ; genetics

Postoperative intra-abdominal adhesion is one of the most common complications in the postoperative period. Current remedies are very ineffective to prevent the pathological outcomes except steroid hormones. Rhynchophylline is deemed as a pharmacologically active component from traditional Oriental medicine Uncaria rhynchophylla (Miq.) Jacks. (Rubiaceae). This study was designed to investigate the preventative effect of rhynchophylline on the abdominal adhesions in rats. Rhynchophylline relieved the experimental abdominal adhesion and decreased the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the blood serum in a dose-dependent manner. The levels of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) were reduced significantly in the peritoneal fluid. The potential mechanism of the activity is related to inhibition of the TGF-β1/Smad signaling pathway.

OBJECTIVETo investigate the effect of Fructus psoraleae on the differentiation of cultured osteoblast MC3T3-E1 cell isolated from neonatal mouse's calvarium.

METHODF. psoraleae preparation was extracted with distilled water. A mouse osteoblast cell line MC3T3-E1 was used as a cell model for screening potency. Cultured MC3T3-E1 osteoblasts were divided into 4 groups: control and F. psoraleae extract 0.02, 0.2, 2 (crude drug) g x L(-1), change the medium and extract every 3 days. The content of ALP and type I collagen were measured. The expression of ALP, type I collagen and osteocalcin mRNA in MC3T3-E1 cell was detected by real-time PCR.

RESULTThe activity of ALP was stimulated by extract of F. psoralea at doses 0.2 g x L(-1), and the content of type I collagen was encouraged at doses 0.2 g x L(-1) . The extract of F. psoralea enhanced the expression of ALP, type I collagen and osteocalcin mRNA in MC3T3-E1 cell. The expression of ALP mRNA was enhanced by extract of F. psoralea at doses 0.2 g x L(-1) for 7-14 d, and the expression of type I collagen and osteocalcin mRNA was enhanced at doses 0.2 g x L(-1) for 14-21 days.

CONCLUSIONF. psoralea can stimulate the differentiation of osteoblasts in vitro. It will offer a reference for the active mechanism research.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Cornus ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Mice ; Osteoblasts ; drug effects ; pathology

Angelicae Sinensis Radix, as a medicinal and edible Chinese medicinal herb, is widely used in clinical practice. It is mainly cultivated in Minxian, Tanchang, Zhangxian and Weiyuan counties of Gansu province. In recent years, with the comprehensive and in-depth study of Angelicae Sinensis Radix in China and abroad, its chemical composition, pharmacological effects and application and development have attracted much attention. In this study, the chemical composition, traditional efficacy, and modern pharmacological effects of Angelicae Sinensis Radix were summarized. On this basis, combined with the core concept of quality markers(Q-markers), the Q-markers of Angelicae Sinensis Radix were discussed from the aspects of mass transfer and traceability and chemical composition specificity, availability, and measurability, which provided scientific basis for the quality evaluation of Angelicae Sinensis Radix.
Angelica sinensis/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Plant Roots/chemistry* ; China

Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.
Breast Neoplasms/enzymology* ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Humans ; MCF-7 Cells ; N-Terminal Acetyltransferases/metabolism* ; Organoplatinum Compounds/pharmacology* ; Oxaliplatin/pharmacology* ; X-ray Repair Cross Complementing Protein 1

OBJECTIVETo investigate the effects of mannan-binding lectin (MBL) on the maturation and cytokine secretion of human dendritic cells (DC) induced by Candida albicans (C. albicans).

METHODSThe plastic-adherent mononuclear cells were prepared from the blood of healthy adult volunteers. The human peripheral blood mononuclear cells-derived dendritic cells (MNC-DC) were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4, and then cultured for 2 days in presence or absence of C. albicans at varying concentration of human MBL ranging from 1 to 20 mg/L. DC's shape and characters were observed under inverted microscopy, the expression of CD83 and CD86 on DC was analyzed by FACS. The levels of TNF-α and IL-6 were detected by ELISA. FACS also was used to investigate the interaction of MBL with immature DC(imDC) and C. albicans. Western blot was used to detect C. albicans-induced IκBα phosphorylation and p65/NF-κB translocation in DC.

RESULTSMBL at higher concentrations (10-20 mg/L) down-regulated the expression of CD83 and CD86 on the monocyte-derived dentritic cells(MoDC) induced by C. albicans, and inhibited the production of TNF-α and IL-6 induced by C. albicans. FACS showed that MBL could not only bind to C. albicans but also bind to imDCs in a Ca2+-dependent manner. Western blot showed that MBL could decrease the phosphorylation of IκBα and the nuclear translocation of p65/ NF-κB.

CONCLUSIONMBL may inhibit TNF-α and IL-6 production induced by C. albicans in DC through NF-κB signaling pathways, suggesting that MBL can play some roles in the regulation of C. albicans-induced immune response.

Candida albicans ; Cell Differentiation ; Cytokines ; Dendritic Cells ; Humans ; Mannose-Binding Lectin ; NF-kappa B ; Protein Transport