AIM: To explore the effect of Buddleia flavonoids drug-containing plasma and androgen receptor(AR) blocker on the expression of STAT1 phosphoprotein.METHODS: In vitro lacrimal gland epithelial cells were cultivated with H2O2 to establish the dry eye apoptosis state. Blank plasma group, Buddleia officinalis plasma total flavonoids interfere with drug-containing group, and the intervention group of testosterone propionate were set. The expressions of STAT1 phosphoprotein of each group were observed by Western blot and AR blocker flutamide was used to explore the intended androgen effect of Buddleia flavonoids.RESULTS: After the intervention of drug-containing plasma, the expression of STAT1 Phosphoprotein in Buddleja officinalis drug-containing plasma intervention group(0.353±0.494) and testosterone propionate intervention group(0.502±0.036) were enhanced and the differences between the two groups were significant (P<0.01). After using the AR blocker in all groups, the expression of STAT1 phosphoprotein in each group (0.268±0.061,0.283±0.106,0.213±0.071) had no difference.CONCLUSION: Buddleja officinalis drug-containing plasma total flavonoids can promote the expression of STAT1 phosphorylation.

OBJECTIVETo investigate the effect of Luyuan Capsule (LYC) leachate on the proliferation and maturation of dendritic cells (DCs) of adult peripheral blood.

METHODSTotally 100 mL peripheral blood was collected from 4 healthy males. The peripheral blood mononuclear cells were isolated using density gradient centrifugation. The precursor DCs were obtained using wall adherent culture. The immature DCs with recombinant human GM-CSF (rhGM-CSF) and recombinant human IL-4 (rhlL-4) added were harvested for 7 days to get immature DCs. The immature DCs cultured with 1% LYC leachate for two days were taken as the experimental group, while the immature DCs cultured with 20 ng/mL recombinant human TNF-alpha (rhTNF-alpha) for two days were taken as the control group. The morphology of DCs was observed by inverted phase contrast microscope, and the cell proliferation was detected by cell counting. The DCs specific marker CD83 was analyzed using flow cytometry.

RESULTSThe proliferation capacities of DCs were improved in the experimental group. More and longer dendritic protrusions and possess were shown on the surface of DCs in the experimental group than in the control group. The CD83 expressions was obviously higher in the experimental group (84.1% +/- 0.3%) than in the control group (58.7% +/- 0.2%), showing statistical difference (P < 0.05).

CONCLUSION1% LYC leachate could promote the maturation and proliferation of peripheral blood DCs, stronger than that of 20 ng/mL rhTNF-alpha.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

Autophagy, an evolutionarily conserved process by which components of the cell are degraded in lysosomes, may facilitate survival of cancer cells under stress conditions. 8-Azaguanine (8-AG), an inhibitor of purine nucleotide biosynthesis, shows antineoplastic activity in multiple tumor cells. However, chemoresistance has restricted its development as an anticancer agent, and the mechanism of 8-AG resistance is not fully understood. We report here that 8-AG induces a protective autophagy to eliminate its cytotoxicity, and inhibition of autophagy increases cellular sensitivity of cancer cells to 8-AG treatment. Using HepG2 or SMMC-7721 hepatic cancer cell lines, we found that 8-AG inhibited cell viability and induced intrinsic apoptosis, accompanied by the up-regulation of the pro-apoptotic protein BimS, one of Bim (also known as BCL-2-like protein 11, BCL2L11) isoforms. Furthermore, 8-AG treatment enhanced the autophagy flux by promoting the dephosphorylation and activation of Unc-51-like autophagy activating kinase 1 (ULK1) via Akt/mTORC1 (mammalian target of rapamycin complex 1) signaling inhibition. Depletion of autophagy-related gene 7 (ATG7) markedly enhanced the level of BimS, and promoted cell death in response to 8-AG. 8-AG in combination with autophagy inhibitor chloroquine (CQ) or bafilomycin A1 (Baf A1) promoted the 8-AG-induced apoptosis in hepatic cancer cells. Altogether, these findings suggest that autophagy promotes chemoresistance of cancer cells for 8-AG, and blocking autophagy increases cellular sensitivity of cancer cells to 8-AG treatment.

Objective To explore the effects of electroacupuncture at Baihui (DU20) and Shenting (GV24) on Alzheimer's disease, and possible mechanism for it.Methods A total of 24 eight-month-old APP/PS1 male mice were randomly divided into model group (n = 8), electroacupuncture group (n = 8) and non-acupoint group (n = 8), and other eight wild-type mice were as wild-type group.The electroacupuncture group accepted electroacupuncture at Baihui and Shenting, while the non-acupoint group accepted electroacupuncture at bilateral subcostal non-acupoint area, and the wild-type group and the model group accepted the same grasping and fixing, for 28 days. Then they assessed with Morris water maze test. The levels of β-amyloid protein (Aβ) and β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in cerebral cortex were detected with immunohistochemistry and immunofluorescence respectively, and the level of BACE1 m RNA with RT-PCR.Results Compared with the model group, the escape latency decreased in the electroacupuncture group (P ＜ 0.001), and the times crossing platforms increased (P ＜ 0.001), while the expression of BACE1 and Aβ decreased (P ＜ 0.001).Conclusion Electroacupuncture may improve the learning-memory ability by inhibiting the expression of BACE1 in the cerebral cortex of APP/PS1 mice to decreasing the level of Aβ.

OBJECTIVETo evaluate the effect on increasing bone cement-bone interface micro-gomphosis intensity with bone cement oscillator.

METHODSOne hundred femoral bones of adult pig were randomly divided into 6 groups: oscillating group (A1) and control group (A2) of anti-tensile force, oscillating group (B1) and control group (B2) of anti-pressure (n = 20 in each group), oscillating group (C1) and control group (C2) of imaging (n = 10 in each group). Mechanics and CT test was performed, micro-gomphosis intensity of bone cement-bone interface between oscillating group and control group was compared.

RESULTSMechanics and CT test showed bone cement-bone interface micro-gomphosis intensity in oscillating group was significantly stronger than control group (P < 0.01).

CONCLUSIONBone cement oscillator can significantly increase micro-gomphosis intensity of bone-cement interface, and reduce long-term aseptic loosening of artificial prostheses.

Animals ; Bone Cements ; Cementation ; Equipment Design ; Femur ; Joint Prosthesis ; Male ; Materials Testing ; Mechanics ; Random Allocation ; Swine ; Vibration

OBJECTIVETo investigate the protective effects of putative AGEs (advanced glycation endproducts) inhibitor salidroside against aging in an accelerated mouse aging model induced by D-galactose.

METHODSA group of 5-month-old C57BL/6J mice were treated daily with D-galactose, D-galactose combined with salidroside, salidroside alone, and control buffer for 8 weeks. At the end of the treatment, serum AGEs levels, neurological activities, expression of glial fibrillary acidic protein (GFAP) and neurotrophin-3 (NT-3) in the cerebral cortex, as well as lymphocyte proliferation and IL-2 production were determined.

RESULTSD-galactose induced mouse aging model was developed as described before. As expected, salidroside blocked D-galactose induced increase of serum AGEs levels. It also reversed D-galactose induced aging effects in neural and immune system, as evidenced by improving motor activity, increasing memory latency time, and enhancing lymphocyte mitogenesis and interleukin-2 (IL-2) production. Furthermore, elevated expression of GFAP and NT-3 in the aged model mice was also reduced upon salidroside treatment.

CONCLUSIONSalidroside inhibits AGEs formation in vivo, which at least partially contributes to its anti-aging effect in D-galactose induced aging model.

Aging, Premature ; blood ; chemically induced ; prevention & control ; Animals ; Cerebral Cortex ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Galactose ; Glial Fibrillary Acidic Protein ; Glucosides ; pharmacology ; therapeutic use ; Glycation End Products, Advanced ; blood ; Interleukin-2 ; metabolism ; Memory ; drug effects ; Mice ; Mice, Inbred C57BL ; Motor Activity ; drug effects ; Nerve Growth Factors ; metabolism ; Nerve Tissue Proteins ; metabolism ; Phenols ; pharmacology ; therapeutic use ; Spleen ; drug effects ; immunology ; T-Lymphocytes ; drug effects

OBJECTIVETo study whether Lycium barbarum glycopeptide 3 (LBGP3) affects T cell apoptosis in aged mice.

METHODSLBGP3 was purified with DEAE cellulose and Sephadex columns. Apoptotic "sub-G1 peak" was detected by flow cytometry and DNA ladder was resolved by agarose gel electrophoresis. Levels of IFN-gamma and IL-10 were measured with specific kits and mRNA expression was detected by RT-PCR. Apoptosis-related proteins of FLIP, FasL, and Bcl-2 were determined by Western blotting.

RESULTSLBGP3 was purified from Fructus Lycii water extracts and identified as a 41 kD glycopeptide. Treatment with 200 microg/mL LBGP3 increased the apoptotic rate of T cells from aged mice and showed a similar DNA ladder pattern to that in young T cells. The reversal of apoptotic resistance was involved in down-regulating the expression of Bcl-2 and FLIP, and up-regulating the expression of FasL.

CONCLUSIONLycium barbarum glycopeptide 3 reverses apoptotic resistance of aged T cells by modulating the expression of apoptosis-related molecules.

Aging ; immunology ; Animals ; Apoptosis ; drug effects ; Fas Ligand Protein ; immunology ; Glycopeptides ; pharmacology ; Interferon-gamma ; genetics ; immunology ; Interleukin-10 ; genetics ; immunology ; Lycium ; chemistry ; Mice ; Mice, Inbred C57BL ; Proto-Oncogene Proteins c-bcl-2 ; immunology ; RNA, Messenger ; immunology ; T-Lymphocytes ; drug effects ; immunology

BACKGROUND:Due to limited sources,poor hemocompatibility and poor anticoagulation performance,small-diameter tissue-engineered blood vessels cannot be applied in clinical practice.OBJECTIVE:To explore the physicochemical and mechanical properties of sheep carotid arteries after the decellularization in order to find appropriate materials for the preparation of tissue-engineered blood vessels.METHODS:Fresh carotid arteries from sheep were randomly divided into two groups:control group,in which,the sheep carotid arteries were cryopreserved for use after trimming and cleaning;experimental group,in which,after trimming and cleaning,the carotid arteries were decallularized by Triton X-100.sodium deoxycholate and EDTA for 24 hours,rinsed for 72 hours,digested with RNA/DNA enzymes for 24 hours,rinsed for 24 hours and reserved for later use.In both groups,blood samples were subjected to hematoxylin-eosin staining,collagen fiber staining,elastic fiber dyeing,and electron microscopy observation.The physical and chemical properties of the blood vessels are tested by tensile strength,wall tension and thickness.RESULTS AND CONCLUSION:(1) The collagen fibers in both two groups were neat and compact in alignment,with no obvious fracture.(2) Hematoxylin-eosin staining showed that:in the control group,the nuclei were distributed in the inner membrane,middle lamella and outer membrane of the vessels,and the fibers ran regularly;in the experimental group,the fibers ran in order but loosely,and there were no nuclei in the inner membrane,middle lamella and outer membrane of the vessels.(3) Elastic fibers in the control group were regular in alignment and mainly distributed in the middle lamella and outer membrane of the vessels,while in the experimental group,the elastic fibers ran regularly but loosely,and mainly distributed in the middle lamella and outer membrane of the vessels.(4) Under the scanning electron microscope,the originally formed vessels were observed in the experimental group,with no cell residues,and the collagen fibers ran orderly with no fracture and with uniform pore structure.(5) The vessel thickness was lower in the experimental group than the control group (P ＜ 0.01),but the tensile strength showed no difference between the two groups,which was 46.55 kPa in the two groups.To conclude,the decelluarized sheep carotid artery can retain the necessary mechanical properties of the blood vessels after achieving the maximum removal of antigenicity.

Objective To explore the effect of electroacupuncture at Baihui (DU20) and Shenting (DU24) acupoints on learning and memory ability in rats with middle cerebral artery occlusion and reperfusion (MCAO/R), and the potential mechanisms. Methods A total of 90 healthy male Sprague-Dawley rats were randomly divided into sham group (n=18) and operation group (n=72). The MCAO/R model was established by suture method in the operation group. Finally, 54 qualified rats of the operation group were randomly divided into ischemia group (n=18), electroacupuncture group (n=18) and non-acupoint group (n=18). The electroacupuncture group received electroacupuncture at Baihui (DU20) and Shenting (DU24) for 14 days. The cerebral infarction volume was measured by magnetic resonance imaging (MRI). The learning and memory ability was tested by Morris water maze. The protein expression of 5-hydroxytryptamine 1A (5-HT1A) receptor was detected by immunohistochemistry. Results There was no significant difference in the cerebral infarction volume among three groups before intervention (F=1.678, P>0.05). Compared with the ischemia group and the non-acupoint group, the cerebral infarction volume signif-icantly reduced (P<0.001);the latency significantly shortened (P<0.001) and the times crossing the platform decreased (P<0.05);the expres-sion of 5-HT1A receptor decreased in the left hippocampus (P<0.05) in the electroacupuncture group after intervention. There was no signifi-cant difference in all the indices between the non-acupoint group and the ischemia group after intervention (P>0.05). Conclusion Electroacu-puncture at Baihui (DU20) and Shenting (DU24) could effectively increase the learning and memory ability of MCAO/R rats, which might relate with inhibiting the expression of 5-HT1A receptor in hippocampus.

OBJECTIVETo explore whether there are gene mutations of Tolloid-like 1 (TLL-1) gene in Chinese patients with sporadic congenital heart disease (CHD).

METHODSOne hundred and fifteen patients with sporadic CHD were selected as CHD group. One hundred and two age and gender-matched healthy people were recruited as control group. After amplifying the exon 10 of the TLL-1 gene by polymerase chain reaction, the polymerase chain reaction products were purified, sequenced and analyzed in order to investigate the TLL-1 gene mutation.

RESULTSAn insertion mutation of base A in the exon 10 of TLL-1 gene was identified in 7 out of 115 CHD patients, including 3 patients with atrial septal defect, 2 patients with ventricular septal defect, 1 patients with patent ductus arteriosus and 1 patients with complex CHD, the total mutation rate was 6.1% in CHD group and 0 in control group (P < 0.01).

CONCLUSIONSTLL-1 gene mutation with an insertion mutation of base A in exon 10 is often in Chinese patients with various CHD. The underlying pathogenesis between TLL-1 gene mutation and occurrence of congenital heart disease in Chinese people remains unclear and warrants further investigations.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; Exons ; Female ; Heart Defects, Congenital ; genetics ; Humans ; Male ; Middle Aged ; Mutagenesis, Insertional ; Pedigree ; Tolloid-Like Metalloproteinases ; genetics ; Young Adult