To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Decidua/metabolism ; Embryo Implantation/*physiology ; Endometrium/metabolism ; I-kappa B Proteins/*biosynthesis ; NF-kappa B/*biosynthesis ; Time Factors ; Uterus/*metabolism ; Uterus/physiology

Objective:To analyze and compare the volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus.Methods:The volatile components in vinegar-processed schisandrae sphenantherae fruc-tus and vinegar-processed schisandrae chinesis fructus were extracted by headspace solid phase-microextraction (HS-SPME) and quali-tatively analyzed by GC-MS.Results:Totally 20 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.55%of the total volatile components , and totally 21 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.90% of the total volatile components .Conclusion: The type and content of volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus are quite different , and the study can provide scientific basis for the two traditional Chinese medicinal materials .

Objective To evaluate the anti-tumour effect of 60℃ LipiodoI in the embolization of VX_2 hepatocarcinoma in rabbits.Methods VX_2 carcinoma cells were surgically implanted into the left liver lobe in 30 male New Zealand white rabbits,which were randomly divided into 3 groups by figure and table method with 10 rabbits in each group.Physiological saline,Lipiodol(37℃),and Lipiodol(60℃)were injected in each group via hepatic artery and liver cancer was embolized.The volume of tumour and serum level of aspartate aminotransferase(AST)were observed after one week,and the survival period of VX_2 rabbits was also observed.Results In the group of Lipiodol(60℃),the growth rate of tumour(0.92? 0.21)was significantly lower than that of control group(3.48?1.17)and Lipiodol(37℃)groups (1.69?0.26),respectively(F=34.95,P0.05),but was significantly higher than the control group(68.6?6.6)U/L(t=19.24,P

Objective To reveal the role of inhibitor of nuclear factor kappa B kinase alpha (IKKα) in renal inflammation after renal ischemia-reperfusion (IR) injury and its potential associated mechanism.Methods Ischemia-reperfusion injury models were induced in a total of 24 healthy C57BL/6 male mice.Renal function and histological changes were estimated.The expression and site of IKKα,p52,RelB,IL-10 and IL-18 were determined by immunohistochemistry and Western blotting.After the short hairpin RNA(shRNA)targeting IKKα was injected into renal parenchyma,renal function and protein expressions of IKKα,p52,RelB,IL-10,IL-18 were detected.Results Compared with sham-operated group[Scr(7.30±0.13) μmol/L,BUN (8.39± 0.30) mmol/L],levels of Scr [(29.80± 2.10)μmol/L,(27.00±3.40) μmol/L,(23.00±3.70) μmol/L] and BUN [(9.47±3.50) mmol/L,(11.68 ±4.30)mmol/L,(13.12±2.10) mmol/L] were higher on day 1,3,7 and the injury of kidney was serious in IR injury group.Immunohistochemical expression of both IL-18 and IL-10 were increased.Markedly increased IKKα,p52 and RelB protein expression were noted in experiments from day 1 to day 7 during kidney recovery period,with a peak on day 3 and then decreasing toward baseline after day 7.Compared with IR injury group,low-expression of IKKα by injection of shRNA up-regulated the expression of IL-18 and down-regulated the expression of IKKα,p52,RelB and IL-10.Conclusions The NF-κB pathway is activated and IKKα expression is up-regulated during the kidney ischemiareperfusion injury,low-expression of IKKα may block inflammation resolution via down-regulation of alternative NF-κB pathway family members of both p52 and RelB.

Objective To examine the association of urinary neutrophil gelatinase-associated lipocalin(NGAL) and interleukin 18(IL-18) with acute kidney injury (AKI) in patients after cardiac surgery. Methods Thirty-three patients undergone cardiac surgery were divided into AKI group and non-AKI group according to the AKI criteria. The Scr, urinary NGAL and IL-18 were measured at different time points. Results Nine of 33 patients (27.27%)developed postoperative AKI, and Scr concentration in AKI group reached its peak within 12-48 hours after cardiac surgery. Urinary concentrations of NGAL and IL-18 at 2 h and 4 h after cardiac surgery were significantly higher than those before operation in AKI patients (P<0.01). The urinary concentrations of NGAL at each time point and that of IL-18 at 2 h and 4 h after cardiac surgery in AKI patients were significantly higher than those in non-AKI patients. After correction by urinary creatinine, the differences of NGAL/Ucr and IL-18/Ucr ratios were still significant (P< 0.01). For concentrations of urinary NGAL, IL-18 and ratios of NGAL/Ucr, IL-18/Ucr at 2 h after surgery, sensitivities and specificities were good with cutoff values at 250 μg/L, 250 μg/mmol and 1800 ng/L, 1800 ng/mmol, respectively. Urinary concentration of NGAL at 2 h after cardiac surgery was positively correlated with Scr at 12 h postoperation in AKI group (r=0.638, P<0.05).Conclusions The incidence of AKI in patients after cardiac surgery is quite high. Urinary concentrations of NGAL, IL-18 and ratios of NGAL/Ucr, IL-18/Ucr at 2 h after cardiac surgery are the early diagnostic markers for AKI, among which urinary NGAL/Ucr is the most sensitive one.

Objective To observe the expression of stromal cell-derived factor 1 (SDF-1) in the kidney after ischemic reperfusion injury (IRI),and explore its relationship with macrophage during the IRI kidney.Methods A total of 28 healthy C57BL/6 male mice were used to establish renal IRI model by clamping both pedicles for 35 min followed by reperfusion.Kidney tissue samples were collected at indicated time points.Renal histological changes were estimated.The expression of SDF-1 was determined by immunohistochemistry,ELISA and real-time PCR.After the liposomal clodronate was injected intraperitoneally,the location of CD68 was observed by immunofluorescence.Renal histology and protein expression of SDF-1 were also detected.Results Compared with sham-operated group,classical tubular damage was found in IRI group,accompanied by a large number of inflammatory cells.The expression of total renal SDF-1 peaked on day 1 and decreased to control levels in the following days.SDF-1 in healthy kidney was localized at cortex,but spread to the corticomedullary area of the kidney during IRI.Compared with IRI groups,elimination of macrophage by injection of liposomal clodronate alleviated renal IRI and down-regulated the expressions of CD68 while up-regulating SDF-1.Conclusions SDF-1 expression is up-regulated in IRI kidney and is associated with macrophage.SDF-1 may play a role in the early phase of acute kidney injury and it may be a new marker in diagnosis of AKI.

Objective To evaluate the respiration-induced dosimetric variance in 3DCRT for midthoracic esophageal carcinoma, in order to guide the radiation oncologist to choose the expansion margin. Methods Ten patients with mid-thoracic esophageal carcinoma were scanned by multi-spiral CT simulator respectively in free breathing ( FB), breath-hold after normal inspiration and expiration ( IBH and EBH )with the same scanning range. Then the CT images of three series were transferred to the treatment planning system. The target volume was outlined following the same standard. Plan1 was designed in the images of FB and transported completely to the images of IBH and EBH as Plan2 and Plan3 respectively to observe the dosimetric variance in target volume. Results For GTV, there was a statistical difference only in V100 of the three plans ( H = 6.423, P = 0.040 ) and no significant difference was found in other indexes. For CTV, the V100 and V95 were better in Plan1 (F=3.992, P=0.030; H=9.920, P=0.007) and no significant difference was found in other indexes. While ()TV, the Dmin, V100 and V95 was better in Plan1 ( F = 3.677, P = 0.039; F = 4.539, P = 0.020; H = 6.846, P = 0.033 ) and no significant difference was found in other indexes. There were no significant differences in all the indexes for the spinal cord and lung in the three plans. Conclusions The change in dose distribution was not so much with the standard expansion. It can meet the needs of clinical treatment.

Objective To study the effect of interleukin (IL)-10 knockout (IL-10-/-) on renal repair after renal ischemia-reperfusion injury in mice.Methods Eighteen IL-10-/-mice (KO) aged 8-10 weeks and 18 C57BL/6 wild type mice (WT) aged 8-10 weeks were divided into control group (Sham) and renal ischemia-reperfusion injury (IRI) group.The renal tissue morphology change was observed by Hematoxylin and eosin (HE) staining and Masson staining.The expressions of IL-18, Ki67 and TGF-β1 were detected by immunohistochemistry.The expression of TGF-beta1 and IL-18 were detected by Western blotting.Results Compared with that in WT-IRI group, in KO-IRI group renal pathological damage was more severe, renal interstitial fibrosis was visible, Ki67 expression of renal tubular epithelial cells decreased distinctly (P＜0.01), the expression of TGF-betal increased significantly (P＜0.01).Conclusion Repair slows down significantly after kidney ischemia-reperfusion injury and fibrosis occurs gradually in IL-10-/-mice, eventually progressing to chronic kidney disease.

BACKGROUND: Peroxisome proliferation-activated receptor-gamma (PPARγ) is the member of nuclear receptor superfamily, and closely related with the formation of atherosclerosis.OBJECTIVE: To investigate the relationship between PPARγ C161→T gene polymorphism and coronary atherosclerotic heart disease (CAHD).DESIGN: Randomized controlled experiment SETTING: Department of Cardiology, Tonai Hospital of Huazhong University of Science and Technology; Department of Cardiology, Zhongnan Hospital of Wuhan University; Center for Human Genome Research,Huazhong University of Science and Technology; Department of Internal Medicine, Affiliated Hospital of Hubei University of Traditional Chinese Medicine; Institute of Geriatrics, Dongzhimen Hospital, Beijing University of Chinese Medicine PARTICIPANTS: Totally 203 CAHD patients aged (65±11) years, including 129 males and 74 females, were the inpatients and outpatients of Zhongnan Hospital of Wuhan University and Tonai Hospital of Huazhong University of Science and Technology from June 2002 to December 2005.And 156 cases of them were diagnosed by coronary arteriongraphy, among which 43 patients without coronary artery affection or with coronary stricture ＜ 50%, and 113 patients with coronary stricture ＞ 50 %. While 89 healthy physical examinees of Han race and mean (59±9) years old were enrolled as control group, including 56 males and 33 females. There was no blood relationship between controls and patients.METHODS: The experiment was conducted at Tongji Hospital of Huazhong University of Science and Technology from June 2002 to December 2005. PPARγ C161→T gene polymorphism was determined by polymerase chain reaction and restriction endonuclease fragment length polymorphisms. The radio-immunity technique, coronary angiography and clinical routine biochemical index were applied to analyze the genotypic frequency and allele frequency distributions as well as the relation between clinical data, biochemical index and different genotypes. The risk factors of CAHD were estimated in the patients of different genotypes.MAIN OUTCOME MEASURES: The genotypic frequency and allele frequency distributions, the relation between clinical data, biochemical index and different genotypes, along with the blood lipid, blood glucose, fasting insulin and body mass index (BMI).RESULTS: Totally 103 CAHD patients and 89 controls were involved in the result analysis of gene polymorphism and yielded different gene distribution frequencies.① In control group, "T" allele frequency was 0.213 and "C" allele frequency was 0.787, and in CAHD group, "T" allele frequency was 0.192 and "C" allele frequency was 0.808. There was no significant difference in the genotypic frequency and C, T allele frequencies between two groups (P ＞ 0.05).② The CC genotype was dominant in CAHD patients with coronary artery lesions, and showed significant differences from "T"allele carriers (CT+TT) (P ＜ 0.05). The CAHD risk in the "T" allele carries (OR: 0.56, 95% CI: 0.24-0.63) was much lower than that in the CC homozygote (OR: 1.92, 95% CI: 1.09-2.54).③ Apolipoprotein B in patients with CC genotype was obviously higher than that in patients with "T" allele (CT+TT) (P ＜ 0.05), and there was insignificant difference in the insulin resistance index (P ＞ 0.05).CONCLUSION: There is an important correlation between the substitution of PPARγ C161→T and CAHD, and "T" allele carriers demonstrate a lower risk of CAHD.

OBJECTIVETo explore the molecular mechanism of pain associated with chronic prostatitis and chronic pelvic pain syndrome (CP/CPPS) in the rat model of prostatic inflammation.

METHODSThirty-six male SD rats were equally randomized to an experimental and a control group, the former injected with 50 μl of 3% λ-carrageenan into the ventral prostate to make the model of non-bacterial prostatic inflammation, while the latter with the same volume of sterile saline solution. At 1, 2 and 4 weeks after modeling, the prostate, L6-S1 dorsal root ganglion (DRG) and spinal cord were harvested for examination of the expressions of the nerve growth factor (NGF), transient receptor potential ankyrin 1 (TRPA1), and calcitonin-gene-related peptide (CGRP) by immunohistochemistry and Western blot.

RESULTSThe expressions of NGF, TRPA1 and CGRP in the prostatic tissue were all significantly increased in the experimental group as compared with the control (P <0.05), with a gradual decrease with the prolonging of time (P <0.05). In the L6-S1 DRG and spinal cord, the expressions of NGF, TRPA1 and CGRP exhibited no significant differences between the experimental and control groups at 1 week after modeling (P >0.05) and kept at high levels in the experimental group at 2 and 4 weeks, though not significantly different from those at 1 week (P >0.05). Statistically significant differences were observed in the expressions of the three proteins in the experimental rats among different time points (P <0.05), but not between the two groups at any time point (P >0.05).

CONCLUSIONThe molecular mechanism of CP/CPPS can be evaluated in the rat model of prostatic inflammation established by injecting λ-carrageenan into the prostate. TRPA1 may play an important role in connecting the upstream and down-stream pathways of CP/CPPS-associated pain.

Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Carrageenan ; Chronic Disease ; Chronic Pain ; metabolism ; Ganglia, Spinal ; metabolism ; Humans ; Male ; Nerve Growth Factor ; metabolism ; Pelvic Pain ; metabolism ; Prostatitis ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; TRPA1 Cation Channel ; TRPC Cation Channels ; metabolism