Influence of Carthamus tinctorius L. has been studied on Ca2+ transmembrane influx in rat aorta when its extract was used at 5, 50, 500?g/ml. It was shown that the resting 45Ca influx in rat aorta ring was not altered by C. tinctorius but the 45Ca influx evoked by norepinephrine (1.2 x 10-6 mol/L ) and KCl solution (100mmol/L) was inhibited in a concentration-dependent manner. The results demonstrated that the receptor-operated Ca2+ channel (ROC) and potential-dependent Ca2+ channel (PDC) in cell membrane of smooth muscle could be blocked by C.tinetorus as well as by verapamil.

Objective To investigate if anodal transcranial direct current stimulation (atDCS) of the right cerebellum improves verbal working memory in amnesic persons with mild cognitive impairment (aMCI).Methods Thirty-nine aMCI were randomly divided into an observation and a control group using a random number table.The observation group was given atDCS at 1.2 mA for 20 minutes every day for 5 days,while the control group was provid ed with fake atDCS in the same way.Before and after the treatment,both groups were tested using forward and back ward digit spans,word reading,visually cued sensorimotor tests and finger tapping.Results After the treatment,the forward and backward digit spans of the observation group had improved significantly compared with before the treatment and with the control group's improvements.Significant improvement was observed in the average backward digit span of the control group,but not in their forward digit span after the treatment.No significant differences be tween the two groups were observed in the other measurements before or after the treatment.Conclusion Direct current stimulation of the cerebellum may improve the verbal working memory deficits of aMCI.Further research should be conducted to find the mechanism.

Objective To study the efficacy of movement imagination combined with biofeedback for stroke patients with upper extremity dysfunction. MethodsEighty stroke survivors were recruited and randomly divided into a movement imagination group ( n =40) and a movement imagination combined with biofeedback group ( n =40).Their EMG mean values during wrist dorsiflexion were amplified and calculated.Fugl-Meyer upper extremity function scores (FMAs) and the modified Barthl index (MBI) were recorded before and after 6 weeks of treatment.ResultsAfter 6 weeks of treatment the two groups had significantly higher mean EMG values,FMA scores and MBI scores,but the effects in the combination group were significantly better than those in the simple movement imagination group. ConclusionMovement imagination can be made more effective by combining it with biofeedback for promoting the recovery of stroke patients with upper extremity dysfunction.

We induced embryogenic calli (EC) and non-embryogenic calli (NEC) from flower filaments of Vitis vinifera L. cv. Chardonnay about 10 days before full bloom. The callus were sub-cultured, observed and verified by somatic embryo induction. PCR primers for VvSUC12 and VvSCU27 were designed according to the corresponding sequences in GenBank. After RNA extraction with RNAplant for EC and NEC cell lines, we synthesized the 1st strand DNA for semi quantitative RT-PCR, and normalized the density of the bands against house-keeping gene Actin. The results of 31 cycles semi-quantitative RT-PCR showed that VvSUC12 was highly expressed in both EC and NEC, with higher expression intensity in NEC than in EC, but not reached the significant level; while the expression of VvSUC27 was only detected in EC, and the expression level was significantly lower than that of VvSUC12. We increased the semi-quantitative RT-PCR cycle number to 35 and found that VvSUC27 gene was weakly expressed in NEC, in EC the intensity of the band was increased comparing with 31 cycles, and the expression level was higher than that of NEC. The paper discussed the differential expression of the two sucrose transporters and their relationship with the sucrose in the tissue culture medium.
Culture Techniques ; methods ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genes, Plant ; Membrane Transport Proteins ; biosynthesis ; genetics ; Plant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vitis ; embryology ; genetics ; growth & development ; metabolism

We constructed a His-tagged prokaryotic expression vector of WUSCHEL gene of Arabidopsis thaliana, pET-31b(+)-WUS-His(6). The induction condition of the fusion protein expression in Escherichia coli was optimized. After purified by affinity chromatography, the recombinant WUS protein was resolved by renaturation of gradient urea dialysis, then used as antigen to immune rabbit to prepare polyclonal antibody. The rabbit anti-WUS antibody titer and specificity were analyzed and confirmed by agarose immunodiffusion testing; the antiserum sensitivity was assayed by dot blot and Western blotting. The results showed that the A. thaliana WUS prokaryotic expression vector was successfully constructed, and the optimized protein expression induction condition in E. coli was 0.5 mmol/L IPTG (isopropy-beta-D-thiogalactoside) at 28 degrees C for 10 hours. The purity of the affinity purified protein was higher than 96%, and the prepared polyclonal antibody was with high specificity and sensitivity, it was able to detect protein antigen at ng level.
Amino Acid Sequence ; Animals ; Antibodies ; isolation & purification ; metabolism ; Arabidopsis ; genetics ; Arabidopsis Proteins ; biosynthesis ; genetics ; Chromatography, Affinity ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Homeodomain Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Plant Proteins ; biosynthesis ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Background/Aims: Transmembrane 4 L six family member 1 (TM4SF1) is highly expressed and contributes to the progression of various malignancies. However, how it modulates hepatocellular carcinoma (HCC) progression and senescence remains to be elucidated. Methods: TM4SF1 expression in HCC samples was evaluated using immunohistochemistry and flow cytometry. Cellular senescence was assessed through SA-β-gal activity assays and Western blot analysis. TM4SF1-related protein interactions were investigated using immunoprecipitation-mass spectrometry, co-immunoprecipitation, bimolecular fluorescence complementation, and immunofluorescence. Tumor-infiltrating immune cells were analyzed by flow cytometry. The HCC mouse model was established via hydrodynamic tail vein injection. Results: TM4SF1 was highly expressed in human HCC samples and murine models. Knockdown of TM4SF1 suppressed HCC proliferation both in vitro and in vivo, inducing non-secretory senescence through upregulation of p16 and p21. TM4SF1 enhanced the interaction between AKT1 and PDPK1, thereby promoting AKT phosphorylation, which subsequently downregulated p16 and p21. Meanwhile, TM4SF1-mediated AKT phosphorylation enhanced PD-L1 expression while reducing major histocompatibility complex class I level on tumor cells, leading to impaired cytotoxic function of CD8+ T cells and an increased proportion of exhausted CD8+ T cells. In clinical HCC samples, elevated TM4SF1 expression was associated with resistance to anti-PD-1 immunotherapy. Targeting TM4SF1 via adeno-associated virus induced tumor senescence, reduced tumor burden and synergistically enhanced the efficacy of anti-PD-1 therapy. Conclusions Our results revealed that TM4SF1 regulated tumor cell senescence and immune evasion through the AKT pathway, highlighting its potential as a therapeutic target in HCC, particularly in combination with first-line immunotherapy.

Background/Aims: Transmembrane 4 L six family member 1 (TM4SF1) is highly expressed and contributes to the progression of various malignancies. However, how it modulates hepatocellular carcinoma (HCC) progression and senescence remains to be elucidated. Methods: TM4SF1 expression in HCC samples was evaluated using immunohistochemistry and flow cytometry. Cellular senescence was assessed through SA-β-gal activity assays and Western blot analysis. TM4SF1-related protein interactions were investigated using immunoprecipitation-mass spectrometry, co-immunoprecipitation, bimolecular fluorescence complementation, and immunofluorescence. Tumor-infiltrating immune cells were analyzed by flow cytometry. The HCC mouse model was established via hydrodynamic tail vein injection. Results: TM4SF1 was highly expressed in human HCC samples and murine models. Knockdown of TM4SF1 suppressed HCC proliferation both in vitro and in vivo, inducing non-secretory senescence through upregulation of p16 and p21. TM4SF1 enhanced the interaction between AKT1 and PDPK1, thereby promoting AKT phosphorylation, which subsequently downregulated p16 and p21. Meanwhile, TM4SF1-mediated AKT phosphorylation enhanced PD-L1 expression while reducing major histocompatibility complex class I level on tumor cells, leading to impaired cytotoxic function of CD8+ T cells and an increased proportion of exhausted CD8+ T cells. In clinical HCC samples, elevated TM4SF1 expression was associated with resistance to anti-PD-1 immunotherapy. Targeting TM4SF1 via adeno-associated virus induced tumor senescence, reduced tumor burden and synergistically enhanced the efficacy of anti-PD-1 therapy. Conclusions Our results revealed that TM4SF1 regulated tumor cell senescence and immune evasion through the AKT pathway, highlighting its potential as a therapeutic target in HCC, particularly in combination with first-line immunotherapy.

Background/Aims: Transmembrane 4 L six family member 1 (TM4SF1) is highly expressed and contributes to the progression of various malignancies. However, how it modulates hepatocellular carcinoma (HCC) progression and senescence remains to be elucidated. Methods: TM4SF1 expression in HCC samples was evaluated using immunohistochemistry and flow cytometry. Cellular senescence was assessed through SA-β-gal activity assays and Western blot analysis. TM4SF1-related protein interactions were investigated using immunoprecipitation-mass spectrometry, co-immunoprecipitation, bimolecular fluorescence complementation, and immunofluorescence. Tumor-infiltrating immune cells were analyzed by flow cytometry. The HCC mouse model was established via hydrodynamic tail vein injection. Results: TM4SF1 was highly expressed in human HCC samples and murine models. Knockdown of TM4SF1 suppressed HCC proliferation both in vitro and in vivo, inducing non-secretory senescence through upregulation of p16 and p21. TM4SF1 enhanced the interaction between AKT1 and PDPK1, thereby promoting AKT phosphorylation, which subsequently downregulated p16 and p21. Meanwhile, TM4SF1-mediated AKT phosphorylation enhanced PD-L1 expression while reducing major histocompatibility complex class I level on tumor cells, leading to impaired cytotoxic function of CD8+ T cells and an increased proportion of exhausted CD8+ T cells. In clinical HCC samples, elevated TM4SF1 expression was associated with resistance to anti-PD-1 immunotherapy. Targeting TM4SF1 via adeno-associated virus induced tumor senescence, reduced tumor burden and synergistically enhanced the efficacy of anti-PD-1 therapy. Conclusions Our results revealed that TM4SF1 regulated tumor cell senescence and immune evasion through the AKT pathway, highlighting its potential as a therapeutic target in HCC, particularly in combination with first-line immunotherapy.