Objective To compare the dosimetric and radiobiological differences of helical tomotherapy (HT) and volumetric modulated arc therapy (VMAT) in craniospinal irradiation. Methods The CT images of 15 patients who received craniospinal irradiation in our hospital were selected. The target volumes and organs at risk (OARs) were contoured, and HT and VMAT plans were designed. The dosimetric parameters of the two plans were compared. A Matlab program based on equivalent uniform dose was developed to calculate the normal tissue complication probability (NTCP). The NTCP values of the two plans were compared. Results The homogeneity index of the target volume in the HT group was better than that in the VMAT group, with values of 0.06 ± 0.01 and 0.08 ± 0.24, respectively, and the difference was statistically significant (P=0.03). However, there was no significant difference in the conformity index of the target volume (P>0.05). There were significant differences in key indicators (D_mean, V₅, D_max) of the lungs, liver, lens, and eyeballs between the two groups (P<0.05). Regarding OARs, the NTCP values of the lens, optic chiasm, lungs, and liver in the HT and VMAT groups were as follows: 0.04 ± 0.03 vs. 0.1 ± 0.06 in the left lens, 0.04 ± 0.06 vs. 0.1 ± 0.07 in the right lens, 0.16 (0.14-0.17) vs. 0.21 (0.18-0.24) in the optic chiasm, 3.89 × 10⁻⁴ (2.45 × 10⁻⁴-7.3 × 10⁻⁴) vs. 8.95 × 10⁻⁴ (5.19 × 10⁻⁴-1.75 × 10⁻³) in the lungs, and 3.45 × 10⁻⁸ (6.0 × 10⁻⁹-1.036 × 10⁻⁷) vs. 9.54 × 10⁻⁸ (1.70 × 10⁻⁸-2.056 × 10⁻⁷) in the liver; the HT group was superior to the VMAT group, and all differences were statistically significant (P<0.05). The NTCP values of the heart in the two groups were 1.35 × 10⁻⁸ (6.34 × 10⁻⁹-2.06 × 10⁻⁹) vs. 5.06 × 10⁻⁹ (2.29 × 10⁻⁹-7.9 × 10⁻⁹), significantly lower in the VMAT group than in the HT group (P<0.05). Conclusion HT has high homogeneity and consistency. The two plans have their own advantages in OAR protection. For OARs with no significant differences in physical dosimetry, NTCP results can be used as a reference. Therefore, comparing the dosimetric parameters and OAR NTCP of HT and VMAT plans can help select the optimal clinical treatment strategy.

ObjectiveTo explore the potential mechanism by which Huangqi Baijiang Yiren decoction （HBY） repairs the intestinal mucosal injury in the rat model of ulcerative colitis （UC） via the miR-21/suppressor of cytokine signaling 1 （SOCS1）/Janus kinase 1 （JAK1）/signal transducer and activator of transcription 6 （STAT6） signaling pathway. MethodsSixty SPF-grade male SD rats were randomly assigned into six groups： blank， model， low-dose （3.68 g·kg^-1） HBY， medium-dose （7.35 g·kg^-1） HBY， high-dose （14.5 g·kg^-1） HBY， and mesalazine （0.035 g·kg^-1）， with 10 rats in each group. The rat model of UC was established in other groups except the blank group by 3% dextran sulfate sodium solution. The rats were administrated with corresponding drugs once a day for 7 consecutive days since the 3th day after modeling. The histopathological changes of the colon were observed by hematoxylin-eosin staining， and the Robarts histopathology index （RHI） was scored. Enzyme-linked immunosorbent assay was employed to measure the levels of pro-inflammatory cytokines ［interleukin （IL）-6， IL-18， IL-1β， and tumor necrosis factor-α （TNF-α）］ in the serum. Real-time PCR was employed to determine the mRNA levels of miR-21， SOCS1， JAK1， and STAT6 in the colon tissue. Western blot was employed to determine the protein levels of SOCS1， JAK1， phosphorylated （p）-JAK1， STAT6， p-STAT6， Occludin， and Claudin-1 in the colon tissue. ResultsCompared with the blank group， the model group showed an increase in disease activity index （DAI） （P<0.01）， shortening of colon length （P<0.01）， severe histopathological damage in the colon tissue， and an increase in RHI， rises in serum levels of IL-6， IL-1β， IL-18， and TNF-α （P<0.01）， up-regulation in mRNA levels of miR-21， JAK1， and STAT6 and protein levels of p-JAK1 and p-STAT6 （P<0.01）， and down-regulation in mRNA and protein levels of SOCS1 and protein levels of Occludin and Claudin-1 （P<0.01）. The treatment with HBY reduced the DAI （P<0.01）， alleviated colon shortening and histopathological damage in the colon tissue， decreased the RHI （P<0.01）， lowered the serum levels of IL-6， IL-1β， IL-18， and TNF-α （P<0.01）， down-regulated the mRNA levels of miR-21， JAK1， and STAT6 （P<0.05， P<0.01）， up-regulated the mRNA level of SOCS1 （P<0.05）， up-regulated the protein levels of SOCS1， Occludin， and Claudin-1 （P<0.05， P<0.01）， and down-regulated the protein levels of p-JAK1 and p-STAT6 （P<0.05， P<0.01）. ConclusionHBY may modulate the miR-21/SOCS1/JAK1/STAT6 signaling pathway to suppress inflammatory responses and restore the intestinal mucosal barrier in UC rats.

Objective This study aims to systematically evaluate the protective role of quercetin(QCT),a naturally occurring flavonoid,against oxidative damage in human endometrial stromal cells(HESCs)induced by hydrogen peroxide(H2O2).Oxidative stress,such as that induced by H2O2,is known to contribute significantly to cellular damage and has been implicated in various reproductive health issues.The study is focused on investigating how QCT interacts with specific molecular pathways to mitigate this damage.Special attention was given to the p38 MAPK/NOX4 signaling pathway,which is crucial to the regulation of oxidative stress responses in cellular systems.By elucidating these mechanisms,the study seeks to confirm the potential of QCT not only as a protective agent against oxidative stress but also as a therapeutic agent that could be integrated in treatments of conditions characterized by heightened oxidative stress in endometrial cells.Methods In vitro cultures of HESCs were treated with QCT at different concentrations(0,10,20,and 40 μmol/L)for 24 h to verify the non-toxic effects of QCT on normal endometrial cells.Subsequently,250 μmol/L H2O2 was used to incubate the cells for 12 h to establish an H2O2-induced HESCs injury model.HESCs were pretreated with QCT for 24 h,which was followed by stimulation with H2O2.Then,CCK-8 assay was performed to examine the cell viability and to screen for the effective intervention concentration.HESCs were divided into 3 groups,the control group,the H2O2 model group,and the H2O2+QCT group.Intracellular levels of reactive oxygen species(ROS)were precisely quantified using the DCFH-DA fluorescence assay,a method known for its accuracy in detecting and quantifying oxidative changes within the cell.The mitochondrial membrane potential was determined by JC-1 staining.Annexin Ⅴ/PI double staining and flow cytometry were performed to determine the effect of QCT on H2O2-induced apoptosis of HESCs.Furthermore,to delve deeper into the cellular mechanisms underlying the observed effects,Western blot analysis was conducted to measure the expression levels of the critical proteins involved in oxidative stress response,including NADPH oxidase 4(NOX4),p38 mitogen-activated protein kinase(p38 MAPK),and phosphorylated p38 MAPK(p-p38 MAPK).This analysis helps increase understanding of the specific intracellular signaling pathways affected by QCT treatment,giving special attention to its potential for modulation of the p38 MAPK/NOX4 pathway,which plays a significant role in cellular defense mechanisms against oxidative stress.Results In this study,we started off by assessing the toxicity of QCT on normal endometrial cells.Our findings revealed that QCT at various concentrations(0,10,20,and 40 μmol/L)did not exhibit any cytotoxic effects,which laid the foundation for further investigation into its protective roles.In the H2O2-induced HESCs injury model,a significant reduction in cell viability was observed,which was linked to the generation of ROS and the resultant oxidative damage.However,pretreatment with QCT(10 μmol/L and 20 μmol/L)significantly enhanced cell viability after 24 h(P<0.05),with the 20 μmol/L concentration showing the most substantial effect.This suggests that QCT can effectively reverse the cellular damage caused by H2O2.Furthermore,the apoptosis assays demonstrated a significant increase in the apoptosis rates in the H2O2 model group compared to those in the control group(P<0.01).However,co-treatment with QCT significantly reversed this trend(P<0.05),indicating QCT's potential protective role in mitigating cell apoptosis.ROS assays showed that,compared to that in the control group,the average fluorescence intensity of ROS in the H2O2 model group significantly increased(P<0.01).QCT treatment significantly reduced the ROS fluorescence intensity in the H2O2+QCT group compared to the that in the H2O2 model group,suggesting an effective alleviation of oxidative damage(P<0.05).JC-1 staining for mitochondrial membrane potential changes revealed that compared to that in the control,the proportion of cells with decreased mitochondrial membrane potential significantly increased in the H2O2 model group(P<0.01).However,this proportion was significantly reduced in the QCT-treated group compared to that of the H2O2 model group(P<0.05).Finally,Western blot analysis indicated that the expression levels of NOX4 and p-p38 MAPK proteins were elevated in the H2O2 model group compared to those of the control group(P<0.05).Following QCT treatment,these protein levels significantly decreased compared to those of the H2O2 model group(P<0.05).These results suggest that QCT may exert its protective effects against oxidative stress by modulating the p38 MAPK/NOX4 signaling pathway.Conclusion QCT has demonstrated significant protective effects against H2O2-induced oxidative damage in HESCs.This protection is primarily achieved through the effective reduction of ROS accumulation and the inhibition of critical signaling pathways involved in the oxidative stress response,notably the p38 MAPK/NOX4 pathway.The results of this study reveal that QCT's ability to modulate these pathways plays a key role in alleviating cellular damage associated with oxidative stress conditions.This indicates not only its potential as a protective agent against cellular oxidative stress,but also highlights its potential for therapeutic applications in treating conditions characterized by increased oxidative stress in the endometrium,thereby offering the prospect of enhancing reproductive health.Future studies should explore the long-term effects of QCT and its clinical efficacy in vivo,thereby providing a clear path toward its integration into therapeutic protocols.

OBJECTIVES: Myocardial ischemia reperfusion injury (IRI) occurs occasionally in the process of ischemic heart disease. Sevoflurane preconditioning has an effect on attenuating IRI. Preserving the structural and functional integrity of mitochondria is the key to reduce myocardial IRI. Silent information regulator 3 (SIRT3), a class of nicotinamide adenine dinucleotide (NAD⁺) dependent deacetylases, is an important signal-regulating molecule in mitochondria. This study aims to explore the role of mitochondrial NAD⁺-SIRT3 pathway in attenuating myocardial IRI in rats by sevoflurane preconditioning. METHODS: A total of 60 male Sprague Dawley (SD) rats were randomly divided into 5 groups (n=12): A sham group (Sham group), an ischemia reperfusion group (IR group), a sevoflurane preconditioning group (Sev group, inhaled 2.5% sevoflurane for 30 min), a sevoflurane preconditioning+SIRT3 inhibitor 3-TYP group (Sev+3-TYP group, inhaled 2.5% sevoflurane for 30 min and received 5 mg/kg 3-TYP), and a 3-TYP group (5 mg/kg 3-TYP). Except for the Sham group, the IR model in the other 4 groups was established by ligating the left anterior descending coronary artery. The size of myocardial infarction was determined by double staining. Serum cardiac troponin I (cTnI) level was measured. The contents of NAD⁺ and ATP, the activities of mitochondrial complexes I, II, and IV, the content of MDA, the activity of SOD, and the changes of mitochondrial permeability were measured. The protein expression levels of SIRT3, SOD2, catalase (CAT), and voltage dependent anion channel 1 (VDAC1) were detected by Western blotting. The ultrastructure of myocardium was observed under transmission electron microscope. MAP and HR were recorded immediately before ischemia (T0), 30 min after ischemia (T1), 30 min after reperfusion (T2), 60 min after reperfusion (T3), and 120 min after reperfusion (T4). RESULTS: After ischemia reperfusion, the content of NAD⁺ in cardiac tissues and the expression level of SIRT3 protein were decreased (both P<0.01), and an obvious myocardial injury occurred, including the increase of myocardial infarction size and serum cTnI level (both P<0.01). Correspondingly, the mitochondria also showed obvious damage on energy metabolism, antioxidant function, and structural integrity, which was manifested as: the activities of mitochondrial complexes I, II, and IV, ATP content, protein expression levels of SOD2 and CAT were decreased, while MDA content, VDAC1 protein expression level and mitochondrial permeability were increased (all P<0.01). Compared with the IR group, the content of NAD⁺ in cardiac tissues and the expression level of SIRT3 protein were increased in the Sev group (both P<0.01); the size of myocardial infarction and the level of serum cTnI were decreased in the Sev group (both P<0.01); the activities of mitochondrial complexes I, II, and IV, ATP content, protein expression levels of SOD2 and CAT were increased, while MDA content, VDAC1 protein expression level, and mitochondrial permeability were decreased in the Sev group (all P<0.01). Compared with the Sev group, the content of NAD⁺ in cardiac tissues and the expression level of SIRT3 protein were decreased in the Sev+3-TYP group (both P<0.01); the size of myocardial infarction and the level of serum cTnI were increased in the Sev+3-TYP group (both P<0.01); the activities of mitochondrial complexes I, II, and IV, ATP content, protein expression levels of SOD2 and CAT were decreased, while MDA content, VDAC1 protein expression level, and mitochondrial permeability were increased in the Sev+3-TYP group (all P<0.01). CONCLUSIONS Sevoflurane preconditioning attenuates myocardial IRI through activating the mitochondrial NAD⁺-SIRT3 pathway to preserve the mitochondrial function.
Adenosine Triphosphate/metabolism* ; Animals ; Male ; Mitochondria/metabolism* ; Myocardial Infarction/metabolism* ; Myocardial Reperfusion Injury/metabolism* ; NAD/metabolism* ; Rats ; Rats, Sprague-Dawley ; Sevoflurane/metabolism* ; Sirtuin 3/metabolism* ; Voltage-Dependent Anion Channel 1/metabolism*

OBJECTIVE To investigate chemical properties of a fructooligosaccharide (ABP-50-FOS)separated from Achyranthes bidentata and immune response in mice immunized H1N1 influenza vaccine. METHODS The methods of GPC,CE,IR and NMR were used to study chemical properties of ABP-50-FOS. BALB/c mice were immunized intramuscularly twice with H1N1 influenza vaccine (3 μg)plus ABP-50-FOS(200 μg)each mouse. The serum total antibody titer and its isotypes titers were analyzed by ELISA. The populations of CD4+,CD8+,CD3+and CD19+lymphocytes were deter?mined by flow cytometry. The proliferation activities of spleen T and B lymphocytes were determined with MTT method. The levels of cytokines interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),inter?leukin-4(IL-4),IL-12 and NO were measured by ELISA kits. RESULTS ABP-50-FOS was a fructooli?gosaccharide with moleculer mass 1885 u. Its bone linkages contained 1,2-and 1,6-fructose residues. ABP-50-FOS could induce high specific-IgG,IgG1,IgG2a,IgG2b and IgM titers after immunization with H1N1 influenza antigen twice(P<0.01). ABP-50-FOS significantly elevated the percentage of CD3+,CD4+ and CD8+ spleen lymphocytes and IFN-γ secretions(P<0.01)in vitro. It also stimulated peritoneal macrophage of mice and DC2.4 dendritic cells to produce TNF-αand IL-12p70 respectively (P<0.01). However,ABP-50-FOS inhibited secretions of NO in macrophage. CONCLUSION The fruc?tooligosaccharide ABP-50-FOS separated from A. bidentata can exhibit strong adjuvant activity for H1N1 influenza vaccine.

Objective To study the effect of black granule capsules(BGCs) on growth of transplanted mouse hepatocarcinoma cells, cell proliferation cycle and apoptosis.Methods KM mouse were subcutaneously inoculated with Hepatocarcinoma cells (H22) and randomly divided into the model control group, the positive control group, the low, medium and high does of BGC group after 24h. The positive control group received intraperitoneal injection with 30 mg/kg cyclophosphamide. Mice of BGC groups were intragastricaly with different dosage of BGC (400, 800, 1 600 mg/kg). The model control group received intragastricaly with normal saline. The drugs were administrated once a day for seven days. The tumor inhibition rate was calculated at 24 h after the last administration. Flow cytometry was used to detect changes of cell cycle and apoptosis in harvested H22 tumor cells.Results The group of high does showed significant inhibitory effect on the growth of transplanted H22 tumor cells withthe inhibitory rate 38.78% (male) and 43.57% (female). Compared with model control group, groups of different dosages decreased time of G0-G1 phase (58.06% ± 9.65%, 55.10% ± 5.89%, 61.36% ± 15.95%vs. 74.47% ± 2.63%), increased tiem of Sphase (33.96% ± 11.90%, 32.67% ± 4.04%, 33.89% ± 9.82%vs. 14.37% ± 4.92%), and increased the apoptosis rate (31.12% ± 1.85%, 31.89% ± 2.16%, 40.64% ± 0.55%vs.21.75% ± 2.64%).Conclusion BGC has antitumor effect on mouse hepatocarcinoma H22 tumor cells, and its mechanism was to regulate cell proliferation cycle and induce the apoptosis.

OBJECTlVE To investigate the effect of α-glycan isolated from Isatis indigotica on humoral immunity and cellular immunity functions in mice immunized with H1N1 influenza vaccine. METHODS BALB/c mice were immunized intramuscularly once with H1N1 influenza vaccine ( 3 μg) plusα-glycan ( 100μg) each mouse. The serum total antibody titer and its isotype antibody titer of immu-nized mice were analyzed by ELlSA at 5, 8, 10, 12 and 14 d after injection at vaccine. The proliferation activities of spleen T and B lymphocytes were determined with MTT method. The levels of cytokines interferon-γ( lFN-γ) , tumor necrosis factorα( TNF-α) , interleukin-4( lL-4) and lL-12 were measured by ELlSA kits. The populations of CD4+, CD8+, CD3+ and CD19+ lymphocytes were determined by flow cytometry. Furthermore, the proliferation rate of macrophages was studied with MTT method in vitro. RESULTS The α-glycan from I.indigotica could gradually induce high specific-antibody production 5-14 d after immunization with H1N1 influenza antigen plus theα-glycan in mice compared to immunization with antigen alone ( P<0.01) . After injection of antigen withα-glycan for 5 d, the main lgG isotype was lgM, and the titer levels of total lgG, lgG1 , lgG2a and lgG2b were also significantly raised following 5-14 d after immunization. The α-glycan significantly promoted the spleen T and B lymphocytes proliferation ( growth rate 44.2%and 37.8%) , stimulated the secretion of lFN-γand lL-12 of splenocytes ( P<0.01, P<0.05) , and also promoted lL-4 secretion of thymocytes (P<0.01). The polysaccharide significantly raised the percent age of CD3+T cells ( P<0.01) , CD3+/CD19+ T lymphocytes ( P<0.01) , and CD8+ T cells ( P<0.01) but decreased the percentage of CD4+/CD8+ T lymphocytes compared with antigen alone group ( P<0.01) . Furthermore, the α-glycan exhibited significant effects on the proliferation and TNF-α secre-tion of MH-S macrophages. CONCLUSlON Theα-glycan isolated from I.indigotica can improve humoral and cellular immunity response in mice immunized with H1N1 influenza vaccine.