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This study aimed to evaluate the effects of ingesting yogurt fermented with Lactobacillus delbrueckii subsp. bulgaricus (OLL1073R-1) on the immune function of healthy university men track and field athletes. Study design Randomized, double-blind, placebo-controlled, parallel-group study. A total of 37 track and field athletes aged ≥18 years were randomly assigned into two groups. For 2 weeks, two bottles of yogurt fermented with OLL1073R-1 and Streptococcus thermophilus OLS3059 or placebo sour milk were ingested daily to the participants. During the intake period, a 1-week training camp was held and participants were subjected to strenuous exercise. Natural killer (NK) cell activity, which is the primary endpoint, was significantly lower in the placebo group after ingestion than that at baseline; however, it remained unchanged during the pre-exercise level of the yogurt group. The two-way repeated measures analysis of variance showed an interaction effect in the NK cell activity change (P=0.018) and a significant difference between the groups after the 2-week ingestion (P=0.015). Among the secondary endpoints, cytokines and chemokines levels involved in activating innate immunity maintained or enhanced only in the yogurt group. ALT, LDH, and CK significantly elevated only in the placebo group. Furthermore, amino acid levels were significantly lower in the placebo group after ingestion than that at baseline; however, it remained unchanged during the pre-exercise level in the yogurt group. Consuming yogurt fermented with OLL1073R-1 prevents the decline in immune function associated with strenuous exercise. Additionally, the yogurt may contribute to stable physical condition.
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Objective:To investigate the effect of 1 064-nm Q-switched Nd:YAG laser at different energy settings on cell viability, protease activity and structures of Malassezia furfur. Methods:Cultured standard strains of Malassezia furfur were divided into several groups to be irradiated with 1 064-nm Q-switched Nd:YAG laser at different energies of 0 (control group) , 500, 600, 700, 800 and 900 mJ, respectively. Then, fungal suspensions in the above groups were inoculated onto the Leeming & Notman medium separately. After 7-day culture, the diameter and number of colonies were measured to evaluate the fungal cell viability, the protease activity was measured by using the whole-milk plate medium, and the ultrastructure of Malassezia furfur in each group was observed by transmission electron microscopy. One-way analysis of variance was used for comparisons among multiple groups, least significant difference- t test for multiple comparisons, and Pearson correlation analysis for analyzing correlations of laser energy with colony diameter, number and protease activity. Results:The colony diameter and number both significantly differed among the control group, 500-, 600-, 700-, 800- and 900-mJ groups (colony diameter: 4.05 ± 0.69, 3.76 ± 0.51, 3.28 ± 0.41, 3.09 ± 0.72, 2.54 ± 0.64 and 2.43 ± 0.41 mm, respectively; colony number: 4 787 ± 597, 4 287 ± 761, 1 879 ± 275, 1 082 ± 248 and 209 ± 42, 72 ± 31 colony-forming units, respectively; F = 14.83, 231.85, respectively, both P < 0.05) , and were significantly decreased in the 600-, 700-, 800- and 900-mJ groups compared with the control group (all P < 0.05) . The laser energy was negatively correlated with the colony diameter and number ( r = -0.67, -0.91, respectively, both P < 0.05) . The protease activity significantly differed among the control group, 500-, 700- and 900-mJ groups ( F = 346.60, P < 0.05) , and was significantly lower in the 700- and 900-mJ groups than in the control group (both P < 0.05) . There was a negative correlation between the laser energy and protease activity ( r = -0.94, P < 0.05) . Transmission electron microscopy showed intact fungal structures in the control group, relatively intact fungal structures in the 500-mJ group, and obviously damaged fungal structures in the 600- to 900-mJ groups, and the greater the laser energy, the more severely the fungal structures were damaged. Conclusion:The 1 064-nm Q-switched Nd:YAG laser could affect the cell viability of and protease activity in Malassezia furfur, and damage its structures.
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BACKGROUND: Cells cannot survive in the area 200 µm away from nutrients in vitro. Vascular network construction is crucial for thick tissue and organ regeneration in tissue engineering. Coaxial cell printing provides a new way to construct vascular-like channels in vitro. OBJECTIVE: To optimize the coaxial cell printing performance of bioink and to build the tissue-engineered scaffolds with vascular-like structure. METHODS: The aseptic sodium alginate solution was prepared by intermittent pasteurization and then frozen. Freeze-dried powder of aseptic silk fibroin was prepared from degummed silk and sealed. The thawed sodium alginate solution was added to the silk fibroin protein freeze-dried powder and human umbilical vein endothelial cells were added to prepare the bioink. The outer axis of the biological three-dimensional printer was connected with the bioink, and the inner axis was connected with the crosslinking agent. The scaffolds were prepared by coaxial printing, and performed by optical coherence tomography, scanning electron microscopy observation and tensile test. Coaxial scaffolds were made by freeze-preserved sodium alginate solution for 7 days with human umbilical vein endothelial cells. Coaxial scaffolds were also made by freeze-dried sodium alginate solution for 7 days with human umbilical vein endothelial cells and silk fibroin protein sealed for 6 months. The cell survival rate was detected by dead and alive staining after 24 hours of culture in vitro. Vascular-like scaffolds with series and parallel structures were designed and printed. The cell proliferation was detected after 1, 3, 7, 10, and 14 days of culture. RESULTS AND CONCLUSIONS: (1) The optical coherence tomography showed that the maximum printing height of the bioink was 9 layers and the overall thickness was about 4.4 mm. Scanning electron microscopy showed that the outer wall of hollow fiber-filament of vascular-like scaffolds presented irregular strip-shaped crimp with micron-scale internal connected pore structure, while the inner wall of hollow fiber-filament had denser pore structure. (2) The elastic modulus of silk protein freeze-dried scaffold was higher than that of sodium alginate solution (P < 0.05). (3) The cell survival rate of scaffolds treated with sodium alginate solution for 7 days was (86.7±3.4)%, and that of scaffolds treated with silk protein freeze-dried powder for 7 days was (98.1±1.2)%, indicating that the sodium alginate solution freeze- preserved for 7 days was free of bacteria and the shelf-life of silk protein could be up to 6 months. (4) The proliferation activity of cells cultured with parallel structure for 7, 10, and 14 days was higher than that with series structure (P < 0.05). (5) These results imply that the scaffolds have good biocompatibility and mechanical properties.
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In this study, we examined the effects of Lentinula edodes mycelia (LEM) on fatigue and immune function in healthy subjects. 22 people were randomly assigned in a 1:1 ratio to the LEM ingestion group and the placebo group. Daily oral ingestion of LEM for 4 weeks (600 mg/day) significantly improved the VAS fatigue score compared with before treatment (p<0.01). Additionally, NK cell activity was increased by ingestion of LEM (p<0.01) and was higher level compared to the placebo group (p<0.05). These results indicate that oral ingestion of LEM may improve the feeling of fatigue and immune function.
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Objective: To investigate the effects of silencing P75 neurotrophin receptor (P75NTR) and nerve growth factor (NGF) overexpression on the proliferative activity and ectopic osteogenesis ability of bone marrow mesenchymal stem cells (BMSCs) combined with demineralized bone matrix for heterotopic osteogenesis. Methods: BMSCs of Sprague Dawley (SD) rats were cultured and passaged by adherent isolation method. The third generation BMSCs were transfected with lentivirus mediated P75NTR gene silencing (group B), NGF overexpression gene (group C), P75NTR silencing and NGF overexpression double genes (group D), respectively, and untransfected cells as control (group A). After 7 days of transfection, the expression of fluorescent protein of the target gene was observed by fluorescence microscope; cell counting kit 8 method was used to detect the cells activity for 8 days after transfection; the expressions of P75NTR and NGF proteins in each group were detected by Western blot. The adhesion of BMSCs to demineralized bone matrix (DBM) was observed by inverted phase contrast microscope and scanning electron microscope after transfection of p75NTR silencing and NGF overexpression double genes. After transfection, BMSCs and DBM were co-cultured to prepare 4 groups of tissue engineered bone, which were respectively placed in the dorsal subcutaneous tissue of 8-week-old SD rats to construct subcutaneous ectopic osteogenesis model ( n=6). HE staining was performed at 4 and 8 weeks after operation. ALP staining was used to observe the formation of calcium nodules at 8 weeks after operation. The expressions of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and osteocalcin (OCN) were detected by real-time fluorescent quantitative PCR. Results: At 7 days after transfection, there was no fluorescence expression in group A, red fluorescence expression was seen in group B, green fluorescence expression in group C, and red-green compound fluorescence expression in group D. The fluorescence expression rate of target gene was about 70%. Western blot detection showed that the relative expression of P75NTR protein in groups A and C was significantly higher than that in groups B and D, and the relative expression of NGF protein in groups C and D was significantly higher than that in groups A and B ( P<0.05). With the passage of time, the cell proliferation activity increased in all groups, especially in group D, which was significantly higher than that in group A at 3-8 days ( P<0.05). The results of inverted phase contrast microscope and scanning electron microscope showed that BMSCs could adhere well to DBM. In the subcutaneous ectopic osteogenesis experiment, HE staining showed that at 4 and 8 weeks after operation, the more bone tissue was formed in group D than in the other 3 groups. ALP staining showed that group D had the highest ALP activity and better osteogenic expression. Compared with group A, the relative expressions of Runx2, ALP, and OCN mRNAs in group D were significantly higher than those in group A ( P<0.05). Conclusion: Silencing P75NTR and NGF overexpression double genes co-transfected BMSCs with DBM to construct tissue engineered bone has good ectopic osteogenic ability. By increasing NGF level and closing P75NTR apoptosis channel, it can not only improve cell activity, but also promote bone tissue regeneration.
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Objective@#To observe the effects of different concentrations and doses of urea on the proliferation and apoptosis of human hemangioma endothelial cells, in order to provide evidence for the further mechanism study of urea in the treatment of hemangioma.@*Methods@#Human hemangioma endothelial cells (HemECs) and normal endothelial cells (VE) were cultured in vitro. Cell viability was detected by CCK-8 after invention with different concentrations(40%, 50%, 60%, 70%) and doses(3, 6, 9 μl/ml) of urea. The apoptosis of HemECs was detected by flow cytometry dual-dye and propidium lodide single dye.@*Results@#The viability of HemECs was significantly lower than that of VE under different concentrations and doses of urea (P<0.05). The inhibition rate of 40% urea on HemECs increased with the increase of urea dose (P<0.05), and the inhibition effect was most obvious at 4 h and 12 h. The apoptosis of HemECs increased in a time and dosage dependent manner with the treatment of 40% urea. High dose(9 μl/ml)of 40% urea significantly promoted the apoptosis of HemECs(P<0.05).@*Conclusions@#Low dose of 40% urea significantly inhibited the proliferation of HemECs, and had no significant effect on VE. However, high doses of urea promoted apoptosis of HemECs.
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Objective To observe the fat particles survival situations in autologous transplantation under different conditions of low temperature.Methods Three sample groups of adult Wistar rats were injected with fresh autologous fat particles,which were stored under 80 ℃ for two months and under-196 ℃ for two months respectively,and the volume,parallel histological detection,glucose transport measurement,MTT experiments and other data were compared and analyzed one month and two months after the transplantation.Results One month after transplantation,graft survival rate was higher in Group injected fresh autologous fat particles than that of Group injected autologous fat particles stored under-80 ℃ and Group injected autologous fat particles stored under -196 ℃ (P＜0.05);Two months after transplantation,the differences among three groups had no statistical significance.Before the transplantation,the activity of autologous fat particles stored in -80 ℃ and-196 ℃ were 68.7％ and 62.5％ of fresh autologous fat particles,respectively,two months after cryopreservation.One month after the transplantation,we found that the activities of autologous fat particles were higher in fresh group than that of-196 ℃ and-80 ℃,There was no significant difference between the groups stored in-196 ℃ and-80 ℃.Two months after the transplantation,the activities of autologous fat particles were higher in fresh group than that of-196 ℃ and -80 ℃.Before transplantation,the particles of fat histology showed no significant difference among three groups.One month after transplantation,fat vacuoles and distinct inflammatory infiltration were observed in all three groups,with the group stored in-80 ℃ as the most serious affected.Two months after transplantation,fresh fat particles and Group stored in-196 ℃ showed large quantity of tightly packed intact fat cells,scattered around the central area in fat vacuoles,while Group stored in -80 ℃ still had a large quantity of fat vacuoles,but the fat vacuoles were obviously fused.Conclusions The fat stored at-80 ℃ and-196 ℃ could meet the needs of clinical use in the short time.The fat stored at-196 ℃ shows the best activity.The fat particles stored at-196 ℃ could be used to establish better blood circulation.The heaviest inflammatory reaction occurs in the graft zone one month after fat transplantation and the decrease of transplanted particle activity is the most obvious.
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PURPOSE: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models.METHODS: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit.RESULTS: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4⁺ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group.CONCLUSION: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.
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Body Weight , Chromatography, Supercritical Fluid , Enzyme-Linked Immunosorbent Assay , Ethanol , Flow Cytometry , In Vitro Techniques , Killer Cells, Natural , Macrophages , Models, Animal , Nitric Oxide , Sargassum , WaterABSTRACT
OBJECTIVE: To explore the underlying mechanism of acupoint catgut embedding in improving primary dysme-norrhea (PD) in rats based on functional activities of the neuro-endocrine-immune (NEI) network. METHODS: Forty female rats were equally randomized into blank control, PD model, medication, and acupoint catgut embedding groups. The PD model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st and 10th d, and 0.2 mg/rat from 2nd to 9th d) and oxytocin (2 U/rat, i．p.). Rats of the medication group were treated by intragastric perfusion of fenbid (0.8 mL/rat, 125 mg/100 mL), once daily for 10 days. The catgut embedding was applied to bilateral "Ciliao" (BL 32), "Sanyinjiao" (SP 6) and "Guanyuan" (CV 4) before modeling. The body writhing times in 30 minutes were recorded, plasma β-endorphin(β-EP) content, and prostaglandin E 2 (PGE2) and prostaglandin F 2 α (PGF2α) contents in the uterus tissue were assayed using ELISA, and the activity of natural killer cell (NK cell) in the spleen tissue was detected using 3-(4，5-dimethyl-2-thiazolyl)-2，5-diphenyl-2-H-tetrazolium bromide (MTT) method after isolation and co-culture with K 562 cells. RESULTS: The body writhing times were no-tably more in the model group than in the control group (P0.05). CONCLUSION: The acupoint catgut embedding has a significant efficacy in relieving PD in rats, which may be related to its effect in up-regulating plasma β-EP, uterus PGE2 contents and splenic NK cell activity and in down-regulating uterus PGF2α level.
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Objective@#BG-derived HIV-1 Tat protein from an HIV-associated dementia (HAD) patient was expressed in E. coli BL21(DE3) and purified in order to research the effects on human umbilical vein endothelial cells (HUVECs) activity.@*Methods@#The recombinant plasmid pGEX-KG-tat with HIV-1 tat stored in our laboratory was amplified by PCR. The PCR product was cloned into pET-32a-tat. The recombinant plasmid pET-32a-tat was transfected into E. coli, and Tat protein was expressed in BL21(DE3), which was induced by IPTG. Then it was purified by Ni-chelating chromatography column and gel filtration preloaded column, and identified by SDS-PAGE and Western blot(WB). The concentration was determined by BCA Kit. Different concentrations of Tat were added into HUVECs to detect their effects on cell activity by cck-8.@*Results@#The Tat with high purity was efficiently expressed in BL21 (DE3) and obtained by using the Ni-chelating chromatography column and gel filtration preloaded column. The concentration was 0.47 mg/ml by using BCA Kit. As the concentration of Tat increased, HUVECs activity decreased. There was no significant difference in cells viability between negative control with 100 ng/ml and 200 ng/ml group (P>0.05). There was significant difference in cells viability between negative control with 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group (P<0.05). But the difference between 300 ng/ml, 400 ng/ml, 500 ng/ml and 1000 ng/ml group was not statistically significant (P>0.05).@*Conclusions@#The HIV-1 Tat with biological activity was efficiently expressed in BL21 (DE3), and the activity of HUVECs was significantly decreased when the concentration reached 300 ng/ml.
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Aim To explore the best method of neural stem cellextraction and culture, and provide a technical reference for thebasic research of neural stem cells. Methods Different extractionand culture methods of neural stem cells were compared.The rate of ball of neural stem cells and the stability of neuralstem cells in undifferentiated state were observed by extraction offetal and neonatal rats cortex, using different types of medium,different inoculation density, different culture methods, differentmethods of changing liquid and different passage methods. Neuralstem cells' activities were detected by Varioskan LUX MultimodeMicroplate Reader. Results ① The brain cortex of fetalrats of 14 d had higher proportion of neural stem cells, less othercells and more neurospheres than newborn rats of 24 h. ② Neuralstem cells could be stabilized in undifferentiated state by usingserum-free medium, while most of the neural stem cellswere differentiated into neurons and glial cells by using serummedium. ③ Neural stem cells, seeding at 1. 0 ×109 ·L-1 , hada large number of neurosperes and were in good condition. ④Suspension culture was beneficial to form a stable neurosphereand keep the neural stem cells in an undifferentiated state thanadherent culture. ⑤ The state of neurosphere by changing halfof the medium and adding medium without discarding was betterthan that by replacing all medium. ⑥ The neurospheres couldbe separated into single cells by mechanical blow in primary generationand the second generation of neurospheres cultured in serum-free medium. While the percentage of viable cells in neuralstem cells was the highest digested with stem cell lysates afterthree generations and the neurospheres re-formed were better. ⑦Neural stem cells' activity of 14 d fetal rat in Accutase digestiongroup was significantly higher than that of the other threegroups, and the difference was significant (P <0. 05). ConclusionsNeural stem cells proliferate and divide well, with highrate of ball formation and good neurosphere condition, which canmaintain a stable undifferentiated state by extracting the cerebralcortex of 14 d fetal rats, using serum-free medium, inoculatinginto a 25 cm2 flask at a density of 1. 0 × 109 ·L-1 , and takingthe suspension culture (adding the medium 1 ~ 1. 5 mL every 2~3 d and passage every 6 ~8 d ).
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Lamin B1 is one of the essential members of the nuclear lamina protein family. Its main function is to maintain the integrity of nuclear skeleton, as well as to participate in the cell proliferation and aging by affecting the chromosome distribution. gene expression, and DNA damage repair. The abnormal expression of lamin B1 is related to certain diseases, including neurological diseases [e.g. neural tube defects (NDTs), adult-onset autosomal dominant leukodystrophy (ADLD)] and tumors (e.g. pancreatic cancer). It is also a potential tumor marker as well as drug target. Further research on lamin B1 will help people understand the molecular mechanism of the emergence and development of neural system diseases and tumors, and define a new future in drug target.
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Humans , Cell Nucleus , Gene Expression , Lamin Type B , Physiology , Neoplasms , Nervous System DiseasesABSTRACT
Objective To investigate the immunoregulatory effect of sirolimus on the xenotransplantation with arterial patch. Methods The xenotransplantation of arterial grafts was taken from the wild-type Bama pigs to cynomolgus monkeys. The peripheral blood mononuclear cells of recipient monkeys at 14 days after xenotransplantation (POD14) were selected as subjects. Dimethyl sulphoxide (DMSO) was used in the control group (volume ratio of 1:1 000) and sirolimus was administered in the sirolimus experimental group (final concentration of 0.1 μmol/L and 0.5 μmol/L). The cells were cultured for 1.0 and 5.5 d, respectively. The activity of POD14 cells was evaluated. The DMSO control and sirolimus experimental groups (final concentration of 0.1 μmol/L) were established and cultured for 5.5 d. The quantity of T and B cells in POD14 cells was counted and the expression levels of cytokines and messenger RNA (mRNA) were quantitatively measured. Results Compared with the DMSO control group, the activity of POD14 cells was significantly decreased after sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 1.0 d (P<0.01-0.001). After sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 5.5 d, the activity of POD14 cells was significantly decreased (both P<0.001). Compared with the DMSO control group, the quantity of CD3+CD4+T cells and CD3+CD8+T cells in POD14 cells was significantly reduced after sirolimus treatment at a final concentration 0.1 μmol/L (P<0.05-0.01), whereas the quantity of CD3-CD20+B cells was considerably elevated (P<0.01). Compared with DMSO control group, the levels of interferon(IFN)-γ, interleukin(IL)-2, IL-4, IL-5 and IL-6 in the sirolimus experimental group were significantly down-regulated (P<0.05-0.001). The expression levels of IFN-γ, tumor necrosis factor(TNF)-α, IL-2, IL-4, IL-5 and IL-6 mRNA were significantly down-regulated (P<0.05-0.001). Conclusions Sirolimus inhibits the proliferation of POD14 cells in the recipient monkeys after xenotransplantation with arterial patch. The underlying mechanism is that the sirolimus can reduce the quantity of T cells and suppress the expression and secretion of immune rejection-related cytokines.
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Objective To design a device for studying the effects of shear stress on the activity of blue-green algae.Methods The device was optimized in terms of the weakness of current shear stress devices,such as small volume,unstability,inability for quantitative analysis and short working period.The effective volume of the new device was 400-700 mL,and the error was less than 13.8％.Moreover,this device could produce a quantitative and uniform shear stress field and be continued to follow observation for more than 96 hours.Results By experiment on growth of microcystis aeruginosa,the device was proved to generate shear stress that could significantly affect the activity of microcystis aeruginosa cells.Conclusions This experimental device is an effective equipment for investigating the effects of shear stress on the activity of blue-green algae.
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Objective To design a device for studying the effects of shear stress on the activity of blue-green algae. Methods The device was optimized in terms of the weakness of current shear stress devices, such as small volume, unstability, inability for quantitative analysis and short working period. The effective volume of the new device was 400-700 mL, and the error was less than 13.8%. Moreover, this device could produce a quantitative and uniform shear stress field and be continued to follow observation for more than 96 hours. Results By experiment on growth of microcystis aeruginosa, the device was proved to generate shear stress that could significantly affect the activity of microcystis aeruginosa cells. Conclusions This experimental device is an effective equipment for investigating the effects of shear stress on the activity of blue-green algae.
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Objective To design a device for studying the effects of shear stress on the activity of blue-green algae.Methods The device was optimized in terms of the weakness of current shear stress devices,such as small volume,unstability,inability for quantitative analysis and short working period.The effective volume of the new device was 400-700 mL,and the error was less than 13.8％.Moreover,this device could produce a quantitative and uniform shear stress field and be continued to follow observation for more than 96 hours.Results By experiment on growth of microcystis aeruginosa,the device was proved to generate shear stress that could significantly affect the activity of microcystis aeruginosa cells.Conclusions This experimental device is an effective equipment for investigating the effects of shear stress on the activity of blue-green algae.
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Objective To design a device for studying the effects of shear stress on the activity of blue-green algae.Methods The device was optimized in terms of the weakness of current shear stress devices,such as small volume,unstability,inability for quantitative analysis and short working period.The effective volume of the new device was 400-700 mL,and the error was less than 13.8％.Moreover,this device could produce a quantitative and uniform shear stress field and be continued to follow observation for more than 96 hours.Results By experiment on growth of microcystis aeruginosa,the device was proved to generate shear stress that could significantly affect the activity of microcystis aeruginosa cells.Conclusions This experimental device is an effective equipment for investigating the effects of shear stress on the activity of blue-green algae.
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<p><b>OBJECTIVE</b>To observe the efficacy differences between acupoint catgut embedding combined with ginger-partitioned moxibustion and regular acupuncture on chronic fatigue syndrome (CFS) of spleen-kidneydeficiency syndrome, and to explore its effects on T lymphocyte subsets and activity of NK cell.</p><p><b>METHODS</b>A total of 60 patients with CFS of spleen-kidneydeficiency syndrome were randomly divided into a catgut embedding combined with ginger-partitioned moxibustion (CECGP) group and a regular acupuncture group, 30 cases in each one. The patients in the CECGP group were treated with acupoint catgut embedding combined with ginger-partitioned moxibustion; the acupoint catgut embedding was applied at Guanyuan (CV 4), Shenshu (BL 23), Pishu (BL 20), Zusanli (ST 36), Qihai (CV 6), once a week, while the ginger-partitioned moxibustion was applied at Guanyuan (CV 4), Qihai (CV 6) and Zusanli (ST 36), once every three days for consecutive one month. The patients in the regular acupuncture group were treated with regular acupuncture at Guanyuan (CV 4), Shenshu (BL 23), Pishu (BL 20), Zusanli (ST 36), Qihai (CV 6), once a day, 6 treatments per week (one day for rest) for consecutive one month. The clinical symptom scores, fatigue scale-14 (FS-14), fatigue assessment instrument (FAI), laboratory test results and total effective rate were compared between the two groups before and after treatment.</p><p><b>RESULTS</b>(1) After treatment, the clinical symptom scores, FS-14 and FAI were reduced in the two groups (all<0.05); after treatment, the clinical symptom scores, FS-14 and FAI in the CECGP group were significantly lower than those in the regular acupuncture group (all<0.05). (2) After treatment, the CD/CD, natural killer cell% (NK%), CD%, CD% were all increased in the two groups (all +4<0.05); the CD/CD, CD%, CD% in the CECGP group were significantly higher than those in the regular acupuncture group (all<0.05). (3) After treatment, the total effective rate was 96.7% (29/30) in the CECGP group, which was similar to 93.3% (28/30) in the regular acupuncture group (>0.05).</p><p><b>CONCLUSIONS</b>The acupoint catgut embedding combined with ginger-partitioned moxibustion, which could effectively relieve the symptoms, regulate T lymphocyte subsets and the activity of NK cell, is an effective method for CFS of spleen-kidneydeficiency syndrome.</p>
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AIM:To probe whether CpG oligodeoxyribonucleotides 7909 (CpG ODN7909) combined with Toll like receptor (TLR)9 affected the chemotherapeutic sensitivity of doctaxel (DOC) in human lung cancer A549 and H520 cell lines.METHODS:Sequences of TLR9 siRNAs were designed.A549 and H520 cells were transfected with TLR9 siR-NA by lipofectamine.The expression of TLR9 was detected by Western blot .The cell activity was measured by CCK-8 as-say.The experiments were divided into blank control group , control siRNA group and TLR9 siRNA interference group.The cell cycle distribution and cell apoptosis were analyzed by flow cytometry .The expression of P38 and Bax was determined by Western blot .The cells in each group were exposed to CpG ODN 7909 and/or DOC.RESULTS: In A549 cells and H520 cells, CpG ODN7909 alone had no obvious effect on the cell activity , G2/M phase arrest and apoptosis , but in-creased the protein expression of P 38 and Bax ( P<0.01) .In addition, there was no significant changes of the above inde-xes in CpG ODN7909 treated-TLR9 siRNA group was observed .DOC alone significantly inhibited the cell activity , higher the G2/M phase fractions, apoptotic rates and Bax expression (P<0.01), but didn’t affect the expression of P38 in all 3 groups.Compared with the cells treated with DOC alone , the cells treated with CpG ODN7909 combined with DOC exhibi-ted lower cell activity, higher G2/M phase fractions, apoptosis rates and more Bax expression (P<0.01), showed no sig-nificant change of P38 expression.In addition, there was no significant change of the above indexes in CpG ODN 7909 com-bined with DOC treated-TLR9 siRNA group was observed .CONCLUSION:CpG ODN7909 may enhance the chemothera-peutic sensitivity of DOC in human lung cancer cells by combining with TLR 9.The mechanism might be related to enhan-cing the inhibitory effect and apoptosis of DOC on the cell activity in vitro, arresting the cells at G 2/M phase of the cells .
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Objective To investigate the effect of pretreatment with electroacupuncture at point Neiguan(PC6) on CK and myocardial cell activity in rabbit myocardial ischemia-reperfusion injury and provide an experimental basis for a relatively specific relationship between pericardium meridian points and the heart. Methods Forty New Zealand big ear rabbits were randomly allocated to groups A, B, C, D and E, eight rabbits each. Group A was a sham operation group. Group B was an ischemia-reperfusion model group, in which the left anterior descending coronary artery was occlude for 30 min and reperfused for 60 min. In groups C, D and E, points Neiguan(PC6), Lieque(LU7) and Hegu(LI4) were separately given electroacupuncture stimulation, 20 min every day, at seven days before model making. Creatine kinase (CK) and myocardial formazan contents were compared between the groups. Results There were statistically significant differences in serum CK and myocardial formazan contents between group B, C, D or E and group A (P＜0.01, P＜0.05). There was a statistically significant differences in serum CK content between group C, D or E and group B (P＜0.01). There was a statistically significant differences in myocardial formazan content between groups C and B (P＜0.05). There were statistically significant differences in serum CK and myocardial formazan contents between group D or E and group C (P＜0.01, P＜0.05). Conclusion Electroacupuncture at point Neiguan(PC6) can reduce myocardial ischemia-reperfusion induced injury to myocardial cells to produce a protective effect on the heart.