Animals ; Arrestins ; metabolism ; Hypoxia ; metabolism ; Male ; Prefrontal Cortex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism ; beta-Arrestin 1 ; beta-Arrestins

Objective To compare the dose, clinical efficacy and acute adverse reactions of intensity modulated radiotherapy(IMRT)and three-dimensional conformal radiotherapy(3D-CRT)combined with three-dimensional brachytherapy (3D-BT) in the treatment of concurrent radiotherapy and chemotherapy for advanced stage cervical cancer patients. Methods Data collection was performed from January 2011 to November 2015 in Chinese PLA General Hospital and Inner Mongolia Cancer Hospital.All 89 patients with advanced stage (Ⅱb-Ⅲb) cervical cancer were treated by pelvic radiotherapy and concurrent chemotherapy, 46 cases of them received IMRT and 3D-BT(IMRT group), 43 cases received 3D-CRT and 3D-BT(3D-CRT group),along with cisplatin chemotherapy.The dose accumulation of external beam radiotherapy and 3D-BT was calculated by deformable image registration to analyze clinical efficacy, acute adverse reactions and prognosis of the two groups.Results (1)Dose of radiotherapy:planning target volume(PTV)coverage of IMRT group and 3D-CRT group were respectively(95.4±4.7)% and(95.1±5.1)%, without significant differences (t=0.289, P=0.773). Compared with the patients treated with 3D-CRT, the volumn receiving at least 30 Gy (V30), V50of rectum, colon, bladder and small intestine and V20of bone marrow in the IMRT group were significantly decreased (P<0.05). Regarding the combined dose, the maximum dose (Dmax) and the minimum dose received by the most exposed 2 cm3volume of the analyzed organ(D2CC)of rectum,colon,bladder and small intestine of IMRT group were significantly lower than those of 3D-CRT group (P<0.05). (2) Short-term efficacy: the effective rate of IMRT and 3D-CRT group were respectively 93% (43/46) and 91% (39/43), with no significant differences (χ2=0.237,P=0.626). (3) Acute adverse reactions: compared with 3D-CRT, IMRT could significantly reduce grade 1-2 acute toxicity in gastrointestinal [63%(29/46) vs 84%(36/43)], genitourinary [17%(8/46) vs 37%(16/43)] and hematologic [57%(26/46) vs 79%(34/43)] system (all P<0.05). There were no significant differences of grade 3 acute adverse reactions of gastrointestinal,genitourinary and hematologic system between two groups(all P>0.05). No grade 4 acute adverse reactions were observed. (4) Prognosis: the overall survival rate at 1, 2-year of IMRT and 3D-CRT group were respectively 95.6%,89.1% and 93.1%,86.1%.The progression-free survival rateat 1, 2-year of IMRT and 3D-CRT group were 91.1%, 89.1% and 88.4%, 86.1%, respectively. There were no significant differences in overall survival rate and progression-free survival rate between two groups (P>0.05). Conclusions Compared with 3D-CRT, IMRT combined with 3D-BT has dosimetry advantages based on dose accumulation algorithms by deformable image registration. IMRT could ensure clinical efficacy and significantly reduce the incidence rate of acute toxicities.

Objective To explore the characteristic of fractional amplitude of low frequency fluctuation (fALFF) and the relationship with the severity of depression, suicidal ideation and suicide risk in depression patients with suicidal ideation with resting-state functional magnetic resonance imaging (rs-fMRI). Methods Resting state functional magnetic resonance imaging maps were conducted using fractional amplitude of low frequency fluctuation (fALFF) in 52 depression patients (30 with suicidal ideation and 22 without) and 21 healthy controls (HCs). The severity of depression was evaluat-ed by using Hamilton Depression scale(HAMD). The suicidal ideation, the suicide risk in depression patients with sui-cidal ideation were both assessed by the Beck Scale for Suicide Ideation. The correlation between the fALFF value and the score of HAMD and the Beck Scale for Suicide Ideation was analyzed. Results MRI revealed significant differences in fALFF in the left superior/middle occipital gyrus and the right middle/inferior occipital gyrus (P<0.05, AlphaSim cor-rected)between depression patients with suicidal ideation and the HCs. Compared to the HCs, depression patients with-out suicidal ideation showed a higher fALFF in the left middle occipital gyrus (P<0.05, AlphaSim corrected). MRI re-vealed significant differences in fALFF in the left middle occipital gyrus (P<0.01, AlphaSim corrected)and the right mid-dle occipital gyrus (P<0.01, AlphaSim corrected) between depression patients with suicidal ideation and without. The fALFF of left middle occipital gyrus (r=0.366, P=0.046) and right middle occipital gyrus (r=0.513, P=0.004) were posi-tively correlated with the scores of HAMD, respectively whereas were not correlated with suicidal ideation and suicide risk. Conclusions Depression patients with suicidal ideation have an abnormal spontaneous activity in their left and right middle occipital gyrus. The increased activity in these brain areas are probably associated with the severity of de-pression whereas are not associated with suicidal ideation or suicide risk.

OBJECTIVETo study the optimum extraction parameters and components on ant oil from Polyrhachis vicina.

METHODThe optimum condifious for supercritical CO2 fluid extraction (SFE-CO2), were investigated with orthogonal design, GC-MS was applied for analyzing. The components and their contents in the ant oil were analyzed by GC-MS, and the contents of lead, zinc and manganese in the oil were determined by ICP-AES.

RESULTThe optimum extraction parameters were achieved, temperature of 50 degrees C, pressure of 30 MPa and time of 2 hours. The extracting yield of the ant volatile oil was 11.4% - 14.3%. 51 Constituents were identified including 9-octadecenoic acid, ethyl oleate, cholesterol, n- Hexadecanoic acid, etc, and the content of various constituents was determined by orea normalization. The oil contained unsaturated fatty acid of 64.6%, lead of 0.80 microg x g(-1), zinc of 0.54 microg x g(-1) and manganese of 0.15 microg x g(-1).

CONCLUSIONThe method showes advantages including faster and efficient of extraction, good quality and no solvent residues in the oil.

Animals ; Ants ; chemistry ; Carbon Dioxide ; Chromatography, Supercritical Fluid ; methods ; Fatty Acids, Unsaturated ; analysis ; Gas Chromatography-Mass Spectrometry ; Lead ; analysis ; Manganese ; analysis ; Materia Medica ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Zinc ; analysis

The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.

Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

This study was aimed to explore the effect of cordyceps sinensis enhancing lymphocyte proliferation and surface CD marker expression in simulated microgravity environment. The splenic lymphocytes were separated from mice and cultured in the rotary cell culture system simulated microgravity environment. The cells were treated with different concentration of cordyceps sinensis solution (0, 6.25, 12.5, 25 and 50 µg/ml) for 24, 48 and 72 h respectively, then the cells were harvested, and analyzed for cell proliferation and the expression of cell surface markers (CD4 and CD8). The results showed that under simulated microgravity environment, the lymphocyte proliferation was inhibited. When the concentration of cordyceps sinensis was 25 or 50 µg/ml, the lymphocyte proliferation, CD4 and CD8 expressions all increased, but 50 µg/ml cordyceps sinensis could inhibit the proliferation ability with the time prolonging. It is concluded that the suitable concentration of cordyceps sinensis displayed the ability to enhance the lymphocyte proliferation and CD marker expression in simulated microgravity environment. These results may be valuable for screening drugs which can be potentially against immunosuppression under simulated microgravity.
Animals ; CD4 Antigens ; metabolism ; CD8 Antigens ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cordyceps ; Immune Tolerance ; Lymphocyte Activation ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Polysaccharides ; pharmacology ; Spleen ; cytology ; Weightlessness Simulation

Purpose: This study assessed the correlation between Epstein-Barr virus (EBV) biomarkers and the eighth American Joint Committee on Cancer staging system and the prognostic values of IgG antibodies against replication and transcription activator (Rta-IgG), IgA antibodies against Epstein-Barr nuclear antigen 1, and BamH1 Z transactivator (Zta-IgA) in locoregionally advanced nasopharyngeal carcinoma (NPC) patients. Materials and Methods: Serum EBV antibody levels were measured by enzyme-linked immunosorbent assay in 435 newly diagnosed stage III-IVA NPC patients administered intensity-modulated radiation therapy±chemotherapy. The primary endpoint was progression-free survival (PFS). Results: Rta-IgG and Zta-IgA levels were positively correlated with the N category and clinical stage. Patients with high Rta-IgG levels (> 29.07 U/mL) showed a significantly inferior prognosis as indicated by PFS (77% vs. 89.8%, p=0.004), distant metastasis–free survival (DMFS) (88.3% vs. 95.8%, p=0.021), and local recurrence-free survival (LRFS) (91.2% vs. 98.3%, p=0.009). High Rta-IgG levels were also significantly associated with inferior PFS and LRFS in multivariable analyses. In the low-level EBV DNA group (≤ 1,500 copies/mL), patients with high Rta-IgG levels had significantly inferior PFS and DMFS (both p < 0.05). However, in the high-level EBV DNA group, Rta-IgG levels were not significantly associated with PFS, DMFS, and LRFS. In the advanced T category (T3-4) subgroup, high Rta-IgG levels were also significantly associated with inferior PFS, DMFS, and LRFS (both p < 0.05). Conclusion Rta-IgG and Zta-IgA levels were strongly correlated with the TNM classification. Rta-IgG level was a negative prognostic factor in locoregionally advanced NPC patients, especially those with advanced T category or low EBV DNA level.

This study was aimed to evaluate the effect of lentinan on the immune function of splenic lymphocytes in rotary cell culture system (RCCS) microgravity environment. The splenic lymphocytes from mice were separated and cultured in the normal gravity and the microgravity environments. The cells were treated with lentinan solution (0, 10, 20 and 40 µg/ml). After incubated with lentinan for indicated times (24, 48 and 72 h), the cell proliferation, secretion of cytokine and the expression of cell surface markers were detected by MTT method, ELISA and flow cytometry respectively. The results indicated that lentinan of above mentioned concentrations did not obviously promote the lymphocyte proliferation, but increased the secretion of IL-2 and IFN-γ and enhanced the expression of lymphocyte surface markers CD4 and CD8 in microgravity environment. It is concluded that lentinan has the ability to enhance the lymphocyte immune function in microgravity environment.
Animals ; Cells, Cultured ; Cytokines ; secretion ; Immune Tolerance ; drug effects ; Immunosuppression ; Lentinan ; pharmacology ; Lymphocyte Activation ; drug effects ; immunology ; Lymphocytes ; cytology ; drug effects ; immunology ; Mice ; Mice, Inbred C57BL ; Spleen ; cytology ; Weightlessness Simulation

Aim To explore the core target,key com-ponents and mechanism of Tibetan medicine Rhodiola crenulata in improving cerebral microcirculation based on literature research,network pharmacology,molecu-lar docking and experimental verification.Methods The chemical components of Rhodiola were collected through literature and database,and the potential tar-gets of Rhodiola crenulata were predicted by reverse pharmacophore matching.The related targets of cere-bral microcirculation disorder were obtained and targets were mapping with Rhodiola crenulata.PPI network was constructed and the core targets were screened.The regulatory network of"herb-component-target-dis-ease"was constructed and key components were screened.GO and KEGG enrichment analysis were conducted,and a"Core target-Pathway-Biological Process"network was constructed.Finally,molecular docking validation was carried out,and RT-qPCR and Western blot were used for animal experiments to fur-ther confirm the results of network pharmacology analy-sis.Results A total of 76 active components of Rhodiola crenulata were obtained and corresponding to 285 targets.Altogether 1074 related targets related to cerebral microcirculation disorder were obtained.A-mong them,there were 97 common targets and the main core targets were 6.The key components were 6.The results of molecular docking showed that the bind-ing activity of three key components to the core target was greater than that of the core target protein and its original ligand.The result of RT-qPCR and Western blot demonstrated that Tibetan medicine Rhodiola cre-nulata could significantly reduce the expression of core target CASP3 and AKT1(P＜0.01).Conclusions Tibetan medicine Rhodiola crenulata can improve the cerebral microcirculation disorder through multi compo-nents,multi targets and multi pathways.This study provides an experimental basis for clinical application of Tibetan medicine Rhodiola crenulata to treat cerebral microcirculation disorder.