Objective To investigate the role of CTLA4-Ig and anti-CD40L monoclonal antibody in the acute rejection of pancreaticoduodenal transplantation model of rats.Methods Pancreaticoduo- denal transplantation model was established from the donor F344 rats to the Lewis recipients(diabetes models).The models were divided into 4 groups:groups A,B,C and D with 12 rats in each group. Two days after transplantation,recipients were injected intraperitoneally with saline,CTLA4-Ig(200?g),anti-CD40L monoclonal antibody(200?g),CTLA4-Ig(200?g)combined with anti-CD40L monoclonal antibody(200?g)respectively.On the day 1,4,7,10 after transplantation,the grafts were harvested for histopathological examination and RT-PCR to determine the levels of interleukin (IL)-2,interferon(IFN)-?,IL-4 and IL-10;The blood CD3~+,CD4~+and CD8~+ T cells were detected by flow cytometry.On the day 4 after transplantation,the CD4~+ CD25~+ T cells in the grafts were de- tected by flow cytometry.Results As compared with group A,the severity of the rejection of grafts in groups B,C and D were depressed;Down-regulation of IL-2 was observed in the groups B,C and D, and the levels in group D were lowest.Down-regulation of IFN-7 was detected in the groups B,C and D,but there was no significantly difference between groups D and B or groups D and C.Up-regulation of IL-4 was observed in the groups B and C,and the levels in group D were lower than in groups A,B and C.Up-regulation of IL-10 was observed in groups B and C,and there was significant difference between groups D and B or groups D and C.The CD3~+,CD4~+ and CD8~+ T cells in groups B,C and D were less,but more CID4~+ CD25~+ T ceils in transplanted pancreas were observed,more notably in group D than in group C.Conclusions Combined use of CTLA4-Ig and anti-CD40L monoclonal anti- body can remarkably diminish the severity of the rejection,which might be mediated by altering the balance in Th1/Th2 and increasing the number of CD4~+ CD25~+ regulatory T cells.Co-stimulation blockade with CTLA4-Ig and anti-CD40L monoclonal antibody induction seems to be an attractive strategy to control allograft rejection.

Objective To examine the induction effects of bone marrow mesenchymal stem cells(BMSCs) transfected with bone morphogenetic protein 7 (BMP7) gene differentiating into chondrocytes. Methods We observed the phenotype of cells which were stained with alcian blue and HE climbing to the six pore plate with invert microscope. The glycosaminoglycan (GAG) value in culture medium was detected in control group,BMP7 transfect and culture medium induced groups after 7,14 and 21 days using standard curve method. Standard curve was described using galacturonic-acid as reference substance. The content of collagen Ⅱ was detected by ELISA method. Results HE and Alcian blue staining showed that BMP7 gene transfection group and the group induced by fluid possess the characteristics of chondrocyte. BMP7 induced BMSCs differentiation to chondrocyte which secrete specific protein called collagen Ⅱ and GAG. Content of GAG were (17.1±3.4),(39.5±5.4),(40.8±6.1)mg/L in control group,BMP7 gene transfected group and induced group,collagen Ⅱ were (89.7±14.3),(152.8±14.5),(155.5± 19.3)μg/L in these three groups separately. Comparing with control group,GAG and collagen Ⅱ of BMP7 gene transfected group and culture medium induced group increased obviously(all P < 0.05),but there was no significant difference between BMP7 gene transfeeted group and culture medium induced group (P > 0.05). Conclusion This active protein induces BMSCs differentiating into chondrocyte,in a level similar to that of inducing medium.

Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).
Antineoplastic Agents, Phytogenic ; chemistry ; Benzofurans ; chemistry ; Flavonoids ; chemistry ; Hep G2 Cells ; Humans ; Inhibitory Concentration 50 ; Morus ; chemistry ; Plant Extracts ; chemistry ; Terpenes ; chemistry

This study assessed the clinical application of transvaginal three-dimensional ultrasound (3D TVUS) in the diagnosis of congenital uterine malformation. A retrospective study was performed on 62 patients with congenital uterine malformation confirmed hysteroscopically and/or laparoscopically. The patients were subjected to transvaginal two-dimensional ultrasound (2D TVUS) and 3D TVUS. The accuracy rate was compared between the two methods. The accuracy rate of 3D TVUS was (98.38%, 61/62), higher than that of 2D TVUS (80.65%, 50/62). 3D TVUS coronal plane imaging could demonstrate the internal shape of the endometrial cavity and the external contour of the uterine fundus. It allowed accurate measurement on the coronary plane, and could three-dimensionally show the image of cervical tube, thereby providing information for the diagnosis of some complex uterine malformation. 3D TVUS imaging can obtain comprehensive information of the uterus malformation, and it is superior to 2D TVUS for the diagnosis of congenital uterine malformations, especially complex uterine anomaly.

Curcumin has been reported to possess antitumor activity with low toxicity. However, the clinical application of curcumin has been significantly limited by its instability and poor metabolic property. In order to overcome these limitations and discover novel small molecules with potential antitumor activity, 29 curcumin mimics were synthesized, which were confirmed by 1H NMR and HR-MS, and their cytotoxic property was evaluated against five human cancer cell lines in vitro. Compounds 16, 18 and 19 exhibited good cytotoxic property, their IC50 value were even below 5 micromol x L(-1) to some cancer cell lines, 5-9 times better than curcumin.
Antineoplastic Agents ; chemical synthesis ; pharmacology ; Cell Line, Tumor ; Curcumin ; analogs & derivatives ; chemical synthesis ; pharmacology ; Drug Screening Assays, Antitumor ; Humans ; Inhibitory Concentration 50

OBJECTIVETo verify whether abnormal expression of calcium/calmodulin dependent serine protein kinase (CASK) and inhibitors of differentiation 1 (ID1) exist in Fb of keloid, and to observe the effect of artesunate on two genes.

METHODSFifteen samples of keloid and 12 samples of normal skin tissue (discarded) excised from patients admitted to our hospital were collected. Tissue particle adherent method was used in the primary culture of Fb, and cells from the third to the eighth passage were used for test. Expressions of CASK and ID1 in Fb harvested from both sources were observed with immunofluorescence staining. Fb of keloid were stimulated with artesunate in various concentration for different time, and the median inhibitory concentration (IC50) was determined with the MTT colorimetric assay, which served as the intervention concentration of artesunate. Fb of normal skin were set as normal control group (NC, treated with medium solution). Fb of keloid were divided into scar control group (SC, treated with medium solution) and scar administration group (SA, treated with artesunate in IC50). The cycle and apoptosis of Fb were detected with flow cytometric assay, and the nucleic acid and protein expressions of CASK and ID1 of Fb in each group were determined with RT-PCR and Western blotting. Data were processed with one-way analysis of variance and LSD-t test.

RESULTSExpressions of CASK and ID1 were detected in two kinds of Fb. The concentration of 75 mg/L was selected as the intervention concentration of artesunate. (1) There were statistically significant differences among the three groups in the percentages of cells in G0/G1 phase and G2/M phase (with F values respectively 118.064 and 163.840, P values all below 0.01). The percentage of cells in G0/G1 phase of group SA was (91.4 ± 1.4)%, which was significantly higher than that of group SC and group NC [respectively (80.7 ± 0.3)% and (82.4 ± 0.6)%, with t values respectively 12.740 and 9.872, P values all below 0.05]. The percentage of cells in G2/M phase of group SA was (6.9 ± 0.3)%, which was significantly lower than that of group SC and group NC [respectively (13.7 ± 0.3)% and (12.7 ± 0.8)%, with t values respectively 43.702 and 12.276, P values all below 0.05]. (2) There were statistically significant differences among the three groups in the early and late apoptotic rates (with F values respectively 61.879 and 4710.862, P values all below 0.01). The early and late apoptotic rates of group SA were respectively (7.1 ± 1.0)% and (14.9 ± 0.3)%, which were significantly higher than those of group SC and group NC [with early apoptotic rate respectively (2.6 ± 0.4)% and (2.7 ± 0.3)%, t values respectively 7.974 and 7.767, P values all below 0.05; with late apoptotic rate respectively (2.3 ± 0.3)% and (2.5 ± 0.4)%, t values respectively 72.882 and 69.792, P values all below 0.05]. (3) The mRNA expression of CASK in group SC was 0.658 ± 0.024, and it was lower than that of group NC (1.076 ± 0.008, t = 28.997, P < 0.01) and group SA (0.855 ± 0.008, t = 13.549, P < 0.01). The protein expression of CASK in group SC was 0.067 ± 0.007, and it was lower than that of group NC (0.179 ± 0.015, t = 12.042, P < 0.01) and group SA (0.132 ± 0.010, t = 9.498, P < 0.01). (4) The mRNA expression of ID1 in group SC was 0.416 ± 0.006, which was higher than that of group NC (0.317 ± 0.020, t = 8.299, P < 0.01) and group SA (0.217 ± 0.009, t = 32.417, P < 0.01). The protein expression of ID1 in group SC was 0.789 ± 0.034, and it was higher than that of group NC (0.366 ± 0.029, t = 16.341, P < 0.01) and group SA (0.114 ± 0.006, t = 33.978, P < 0.01).

CONCLUSIONSIt is speculated that CASK and ID1 participate in the proliferation of Fb in keloid. The mechanism of artesunate in inhibiting the proliferation of Fb in keloid may be related to the up-regulation of CASK and down-regulation of ID1.

Adolescent ; Adult ; Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Fibroblasts ; metabolism ; Gene Expression Regulation ; Guanylate Kinases ; genetics ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the possibility of using bioaugmentation as a strategy for remediating quinoline-contaminated soil.

METHODSMicroorganisms were introduced to the soil to assess the feasibility of enhancing the removal of quinoline from quinoline-contaminated soil. Slurry-phase reactor was used to investigate the bioremediation of quinoline-contaminated soil. HPLC (Hewlett-Packard model 5050 with an UV detector) was used for analysis of quinoline concentration.

RESULTSThe biodegradation rate of quinoline was increased through the introduction of Burkholderia pickettii. Quinoline, at a concentration of 1 mg/g soil, could be removed completely within 6 and 8 hours with and without combined effect of indigenous microbes, respectively. Although the indigenous microbes alone had no quinoline-degrading ability, they cooperated with the introduced quinoline-degrader to remove quinoline more quickly than the introduced microbes alone. Bioaugmentaion process was accelerated by the increase of inoculum size and bio-stimulation. The ratio of water to soil in slurry had no significant impact on bioremediation results.

CONCLUSIONBioaugmetation is an effective way for bioremediation of quinoline-contaminated soil.

Biodegradation, Environmental ; Bioreactors ; Burkholderia ; metabolism ; Chromatography, High Pressure Liquid ; Quinolines ; Sewage ; Soil ; analysis ; Soil Microbiology ; Soil Pollutants

OBJECTIVENeonatal asphyxia is one of the main causes for the acute respiratory distress syndrome (ARDS) in full-term newborns. Now it is believed that the reduced amount and abnormal function of pulmonary surfactant due to various causes is a major factor leading to acute lung injury. This study aimed at using an intrauterine acute ischemic-hypoxia rat model and investigating the effect of intrauterine acute ischemic-hypoxia on the expression of surfactant protein A (SP-A) and surfactant protein B (SP-B) in neonatal rat lungs.

METHODSThe rat model of acute intrauterine ischemic-hypoxia was established by ligating the unilateral uterine horn vessels of Wistar rats at the 21st gestational day. While the rat pups from the other side of the uterus, of which the uterine horn vessel was not ligated, were the sham-operation group. Rat pups were delivered by cesarean section at the 20, 30 and 40 min following the ischemic-hypoxia insult. The rat pups delivered by cesarean section from the gestation of 21 days were the normal control group. There were 42 rat pups and 6 pups in each group in this study. The distribution of SP-B protein in the neonatal rat lungs of different period of ischemia was examined by using SABC method. The average gray value of SP-B staining in type II alveolar epithelial cells were measured by Universal Imaging Porporation with Meta Morph software. The reverse transcription polymerase chain reaction (RT-PCR) was performed to quantitate the expression of SP-A and SP-B mRNA.

RESULTSFollowing the intrauterine acute ischemic-hypoxia, the numbers of type II alveolar epithelial cells with the positive SP-B staining were markedly declined. The average gray values at the 20, 30 and 40 min after the ischemia were 78.89 +/- 1.08, 79.69 +/- 0.13 and 80.00 +/- 0.63, respectively, which increased significantly compared with the normal control group (76.13 +/- 0.43, P < 0.01). The expression of SP-A and SP-B mRNA was weak following the ischemic-hypoxia insult. The relative amounts of SP-A (1.16 +/- 0.06, 1.14 +/- 0.01 and 1.13 +/- 0.04, respectively) and SP-B (0.81 +/- 0.02, 0.78 +/- 0.02 and 0.79 +/- 0.04, respectively) at the 20, 30 and 40 min after the ischemia were reduced significantly compared with controls (1.27 +/- 0.09 and 0.89 +/- 0.06, respectively, P < 0.05 and < 0.01) and reduced gradually following the prolongation of the insult. There were no significant differences (P > 0.05) between the normal and sham operation control groups on the expressions of SP-B protein as well as the SP-A and SP-B mRNA.

CONCLUSIONThe reduced synthesis of SP-B protein and the reduced expression of SP-A and SP-B mRNA might be caused by intrauterine acute ischemic-hypoxia, which may support theoretically the early application of pulmonary surfactant including SP-A and SP-B for treating the lung injuries of asphyxia in newborns.

Animals ; Animals, Newborn ; Female ; Gene Expression ; Hypoxia ; physiopathology ; Ischemia ; physiopathology ; Lung ; metabolism ; Pregnancy ; Pulmonary Surfactant-Associated Protein A ; genetics ; Pulmonary Surfactant-Associated Protein B ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Uterus ; blood supply

Objective To investigate the effects of panax notoginseng saponins (PNS) combined with total flavonoids from epimedium (TFE) on testicular dysfunction in natural aging rats; To discuss its mechanism of action. Methods Thirty 18-month old male SD rats were randomly divided into natural aging group, PNS combined with TFE low and high dose groups, with 10 rats in each group. Another 10 2-month old rats were set as young control group. PNS combined with TFE low and high dose groups were given gastric gavage of 10 mg/kg PNS combined with 10 mg/kg TFE, and 20 mg/kg PNS combined with 20 mg/kg TFE, respectively. Rats in young control group and natural aging group were given saline for 6 d each week, lasting for 4 months. Then, rats were sacrificed, and the testes were obtained to calculate the testicular weight and the testicular index. The testicular tissue morphology was observed by using HE staining. Testicular germ cell apoptosis was detected by using TUNEL method. The levels of Bcl-2, Bax and γ-H2 AX protein expression in testicular tissue were detected by Western blot. Results Compared with natural aging group, low and high dose of PNS combined with TFE significantly elevated the testicular weight and testicular index, improved the histological changes of testicular seminiferous tubule, significantly reduced number of apoptosis of spermatogenic cells in the testis, upregulated the expression of Bcl-2 protein in the testis, downregulated the expression of Bax and γ-H2 AX protein, and decreased the ratio of Bax/Bcl-2. Conclusion PNS combined with TFE can improve testicular dysfunction in natural aging rats by inhibiting apoptosis and DNA damage of germ cells.

Objective To observe the anatomic and imaging morphology ofjugnlar bulb and its relationship with the surrounding structures, and to investigate the classification ofjugnlar bulb and its clinical significance. Methods We dissected 30 human temporal bones and studied multi-slice spiral CT imaging data of temporal bone of 120 cases and blood vessel cast mould specimen of the jugular bulb of 6 cases, to observe the morphology of jugnlar bulb and its spatial relationship with the surrounding structures. We made an imagined sagittal plane on the medial well of the tympanic cavity, with a horizontal tangent line of the proximal wall of the tympanic cavity and a vertical tangent line of the posterior wall of the tympanic cavity as coordinate axes (X axis and Y axis), respectively, so the 4 quadrants ( Ⅰ , Ⅱ, Ⅳ, Ⅳ) were formed. The jugular bulb was classified intro 4 types according to the quadrant where its top was projected and subtyped according to its position on the inner or outer side of the plane. The operation via mastoid approach was simulated on specimen to observe the effect of jugnlar bulb on the operation route. Results Some jugular bulbs were flat type and others were prominent types. The classification in the group of CT image: type Ⅰ , 11 case (9%);type Ⅱ, 63 cases (53%);type Ⅲ, 25 cases (21%);type Ⅳ, 21cases (17%). The classification in the group of specimen: type Ⅰ, 1 case (3%);type Ⅱ, 11 cases (37%);type Ⅲ, 8 cases (27%);type Ⅳ, 10 cases (33%). Each type of the jugular bulb had different effects on the operative approach. Conclusions The classification method with the 4 quadrants is a simple and three-dimensional way to describe the position of the jugular bulb for imaging diagnosis or operative scheme design.