Objective:To investigate the structural alterations of mitochondria and its role in neutrophil death induced by ONO-AE-248.Methods:Human neutrophils were cultured in vitro with ONO-AE-248(5?10-5 mol/L)and medium alone. Transmission electron microscopy(TEM) was used to detect the structural alterations of mitochondria and the level of mitochondria membrane potential by flow cytometry using mitocapture dying.Results:ONO-AE-248 resulted in extremely swollen mitochondria within 6 hours. Meanwhile, a rapid loss of mitochondrial membrane potential of neutrophils occurred, especially in 3 hours and 6 hours. There were obviously differences between spontaneous apoptosis and non-apoptosis programmed cell death induced by ONO-AE-248.Conclusion:The experiment results suggest that changes of mitochondrial structure and function be typically morphological, physiological and biochemical features in this unique form of neutrophil death, and that the mitochondrial pathway might play a more important role in ONO-AE-248-induced death of neutrophils.

Lysins are efficient bacteria cell wall digesting enzymes encoded by DNA bacteriophage. Gram-positive bacteriophage lysins feature similar domain structure, high lytic efficiency, synergic antibacterial effect with antibiotics, rare neutralization by antibodies, less chance of developing drug-resistant strains, et al. The past decade has seen a considerable amount of research worldwidely focused on lysin, and lysins have been used successfully in a variety of animal models to control pathogenic antibiotic resistant bacteria found on mucosal surfaces and infected tissues. The great potential of lysins as an anti-infective agent prompted this review.

Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.

Objective:To observe the effects of moxibustion on serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α), and to explore the effects of moxibustion on inflammatory damaging factors in experimental rheumatoid arthritis (RA) model rats; the relationship between the therapeutic effect of moxibustion on RA and the change in the Toll-like receptor (TLR) signaling pathway was analyzed using Toll-like receptor 4 (TLR4) antagonists and agonists. Methods:Fifty Sprague-Dawley (SD) rats were divided into a normal group, a model group, a moxibustion group, a moxibustion plus TLR4 agonist group (agonist group) and a moxibustion plus TLR4 antagonist group (antagonist group) according to the random number table, with 10 rats in each group. Except the normal group, rats in the other four groups were subjected to model preparation with the wind, cold and wet environmental factors plus Freund's complete adjuvant (FCA). Rats in the normal and model groups were not treated; rats in the moxibustion, agonist and antagonist groups started to be treated with the moxibustion (cigarette-type moxa) at bilateral Shenshu (BL 23) and Zusanli (ST 36) from the 4th day after the successful modeling, for 20 min each time with a total of 10 d. Rats in the agonist and the antagonist groups were injected with TLR4 agonist or antagonist [0.1 mg/(kg·bw)] via the tail vein 30 min before moxibustion. The concentrations of serum IL-6, IL-8 and TNF-α in each group were determined by enzyme-linked immunosorbent assay (ELISA). Results:Compared with the normal group, in the model group, the rat's right hind paw swelling was significantly obvious (P<0.01), there was a lot of inflammatory infiltration in the synovial tissues, the surface of the synovial membrane was unsmooth, the synovial membrane was hyperplasia and thicker, and the serum IL-6, IL-8 and TNF-α concentrations increased significantly (P<0.05). Compared with the model group, the paw swelling degrees of the rats in the moxibustion, the agonist and the antagonist groups reduced significantly (allP<0.01); the swelling degree in the antagonist group was milder than that in the agonist group, but the between-group difference was not statistically significant (P>0.05); inflammatory infiltration and synovial membrane hyperplasia in the synovial tissues of the moxibustion group and the antagonist group were all relieved differently; the decrease of synovial layer number in the moxibustion group was more obvious, and there were no obvious improvements in inflammatory infiltration and synovial thickness in the agonist group; the concentrations of IL-6, IL-8 and TNF-α in the moxibustion group were decreased, and the differences in the IL-6 and TNF-α concentrations were statistically significant (allP<0.01); there was no significant between-group difference in the IL-8 concentration (P>0.05); the concentrations of serum IL-8 and TNF-α in the agonist group increased significantly (both P<0.01), while the IL-6 concentration decreased without significant difference (P>0.05); the concentrations of IL-6 and IL-8 in the antagonist group decreased but the between-group differences were statistically insignificant (bothP>0.05), and the TNF-α concentration significantly increased (P<0.05). Compared with the moxibustion group, IL-6, IL-8 and TNF-α concentrations increased in the agonist group, and the differences in the IL-8 and TNF-α concentrations were statistically significant (both P<0.01); the concentrations of IL-6, IL-8 and TNF-α increased in the antagonist group, and the differences in the IL-6 and TNF-α concentrations were statistically significant (bothP<0.01); there was no significant difference in the IL-8 concentration between the groups (P>0.05). The serum levels of IL-6, IL-8 and TNF-α in the antagonist group were lower than those in the agonist group (allP<0.05). Conclusion:Moxibustion at Shenshu (BL 23) and Zusanli (ST 36) can reduce the joint swelling degree and inflammation in synovial tissue of RA model rats, decrease the serum levels of IL-6, IL-8 and TNF-α in RA model rats; the decreases of IL-6 and TNF-α are more significant than the decrease of IL-8; TLR4 agonist and antagonist can significantly attenuate the effect of moxibustion in inhibiting releases of IL-6, IL-8 and TNF-α, so that the change in TLR signaling pathway affects the effect of moxibustion in inhibiting the releases of IL-6, IL-8 and TNF-α.

This study was to investigate the chemical constituents of the aerial part of Zygophyllumfabago, by phytochemical methods. The compounds were isolated by silica gel and Sephadex LH-20 column chromatographies from the EtOAc extract. Their structures were characterized by various spectroscopic data (1H-NMR, 13C-NMR, MS) and comparison with the literature. As a result, thirteen compounds were isolated and their structures were identified as 1-hydroxyhinesol(1), hinesol(2), atractylenolactam(3), beta-eudesmol (4), 5alpha-hydroperoxy-beta-eudesmol(5), 12-hydroxy-valenc-1(10)-en-2-one(6), pubinernoid A(7), (6S,7E)-6-hydroxy-4,7-megastigmadien-3,9-dione(8), 3-hydroxy-5alpha, 6alpha-epoxy-beta-ionone (9), (3S,5R, 6S, 7E)-3, 5, 6-trihydroxy-7-megastigmen-9-one(10), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one(11), (S)-3-hydroxy-beta-ionone(12), and blumenol A(13). Compounds 1-7 were sesquiterpenoids and 8-13 were megastigmane type norsesquiterpenoids. All the compounds were obtained from Z. fabago for the first time, and compound 1 was a new natural product.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Terpenes ; chemistry ; isolation & purification ; Zygophyllum ; chemistry

OBJECTIVETo investigate the intra-vaginal ejaculation latency time (IELT) of varicocele patients, the influence of spermatic vein ligation on IELT, and the relationship of Visual Analogue Score (VAS) with IELT.

METHODSWe selected 112 males who had regular sexual life after spermatic vein ligation and conducted follow-up visits for 6 months. According to preoperative IELT, we divided the patients into an IELT < or = 2 min group and an IELT > 2 min group, and compared their IELT, VAS and Chinese Index of Sexual Function for Premature Ejaculation-5 (CIPE-5) scores before and 6 months after operation.

RESULTSFollow-up was accomplished in 81 of the patients, 18 in the IELT < or = 2 min group and 63 in the IELT >2 min group. Compared with the baseline, IELT was significantly prolonged postoperatively in both the IELT < or = 2 min group ([1.26 +/- 0.37] vs [4.53 +/- 1.69] min, P < 0.01) and the IELT >2 min group ([5.14 +/- 2.03] vs [7.69 +/- 4.51] min, P < 0.05); the postoperative CIPE-5 scores were remarkably improved in the former (11.27 +/- 3.52 vs 15.64 +/- 2.37, P < 0.05) but insignificantly in the latter group (20.42 +/- 4.65 vs 21.83 +/- 5.49, P > 0.05); the postoperative grades of the CIPE-5 scores showed significant differences in both groups (chi2 = 6.353, P = 0.042 and chi2 = 3.910, P = 0.048); the postoperative VAS was markedly increased (3.18 +/- 0.92 vs 1.56 +/- 0.83 and 3.24 +/- 0.95 vs 1.74 +/- 0.79, P < 0.05), with significant differences in the grades of VAS in both groups (chi2 = 4.433, P = 0.035 and chi2 = 10.088, P = 0.001). The variation of VAS was negatively correlated with that of IELT in both groups (r = -0.572, P < 0.01 and r = -0.465, P < 0.05).

CONCLUSIONVaricocele may be one of the causes of premature ejaculation, and some of the varicocele patients with IELT < or = 2 min may benefit from spermatic vein ligation. Improved VAS is negatively correlated with prolonged IELT. The relationship between varicocele and premature ejaculation deserves further studies.

Adult ; Ejaculation ; physiology ; Follow-Up Studies ; Humans ; Ligation ; Male ; Retrospective Studies ; Varicocele ; surgery ; Young Adult

Objective To investigate the effect of Ulinastatin on the delivery of cytokines in patients with septic shock.Methods It was a prospective and controlled clinical study.Seventy-eight patients with septic shock were randomly divided into control group and treatment group and thirty-nine in every group.Patients in treatment group received Ulinastatin 200 000 units intravenous everyday for 3 days,while those in control group received equal volume of normal saline as placebo.At different time points (at 24 th,48 th,72 th hour after start of treatment),the levels of tumor necrosis factor-alpha (TNF-?),interleukin-1 (IL-1),interleukin-6 (IL-6 ),interleukin-8 (IL-8) and superoxide dismutase (SOD) in serum were assayed.Results In comparison with control group,the levels of TNF-?,IL-1,IL-6,IL- 8 of treatment group decreased markedly (P＜0.05,P＜0.01) at different time points,whereas the level of SOD was higher markedly (P＜0.05,P＜0.01) at various time points.Conclusion Ulinastatin has protective effect on patients with septic shock through decreasing the levels of TNF-?,IL-1,IL-6,IL-8 and increasing in the level of SOD.

Aged ; Aged, 80 and over ; Biomarkers, Tumor ; urine ; Carcinoma, Transitional Cell ; urine ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; RNA, Messenger ; urine ; Reverse Transcriptase Polymerase Chain Reaction ; Urinary Bladder Neoplasms ; urine

OBJECTIVETo explore the central mechanism of moxibustion on analgesic effect.

METHODSMale Wistar rats were screened by pain threshold value before making model, and 48 rats whose pain threshold was (250 +/- 25) g were selected. Twelve male Wistar rats were randomly selected as a normal group. For the rest rats the rheumatoid arthritis (RA) model was duplicated by raising in a windy, cold and wet environment combined with injection of Freund's complete adjuvant (FCA), and then they were randomly divided into a model group, a moxibustion group and a moxa volatile oil group, 12 rats in each group. The moxibustion and the moxa volatile oil igroup were treated with moxibustion and moxa volatile oil at "Shenshu"(BL 23) and "Zusanli"(ST 36), respectively, for 15 days. No interventions were added on the model group and the normal group. The pain threshold in Iinjured foot and the expression of hypothalamic POMC mRNA and PDYN mRNA in rats were observed.

RESULTSCompared with the normal group, the pain threshold and the expression of hypothalamic POMC mRNA and PDYN mRNA in the model group were increased (all P < 0.01). Compared with the model group, the pain threshold and the expression of hypothalamic POMC mRNA and PDYN mRNA in the moxibustion group were increased significantly (all P < 0.01), but no statistically significance in the moxa volatile oil group (P > 0.05). Compared with the moxa volatile oil group, the above-mentioned observative indices in moxibustion group were all increased significantly (all P < 0.01).

CONCLUSIONMoxibustion has obvious analgesic effect and its mechanism may be related to the increasing expression of hypothalamic POMC and PDYN mRNA through the warming effect of moxibustion.

Animals ; Arthritis, Rheumatoid ; genetics ; metabolism ; therapy ; Enkephalins ; genetics ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Moxibustion ; Pro-Opiomelanocortin ; genetics ; metabolism ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the value and dynamic characteristics of diffusion-weighted imaging(DWI)in rabbit liver VX_2 tumor model,with correlation of pathology.Methods Forty New Zealand rabbits were included in the study and VX_2 tumor piece was implanted directly into the liver after laporotomy.Fiffteen had two intrahepatic implants while twenty-five had one implant.DWI was performed on the seventh,fourteenth and twenty-first day after implantation,while routine T_1WI and T_2WI sequences were done on the seventh and fourteenth day.Ten VX_2 tumor samples were studied by pathology.Results The lump detection rates on the seventh day after implantation of DWI,T_1WI and T_2WI were 78.7%(37/47), 10.7%(3/28)and 53.6%(15/28)respectively with statistical significance(x~2=32.61,P