Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P＜0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.

In order to investigate the role of MMP-9 and TIMP-1 in the pathogenesis of systemic sclerosis, the expression of MMP-9 and TIMP-1 was immunohistochemically detected in skin lesions of the patients with diffuse cutaneous systemic sclerosis, skin lesions of the patients with limited cutaneous systemic sclerosis, and skin tissues of normal subjects. The results showed that the expression of MMP-9 in lesions of diffuse cutaneous systemic sclerosis was significantly lower than that of normal skins (P<0.05). However, no significant difference in the level of MMP-9 in the limited cutaneous systemic sclerosis and normal skin was found. Meanwhile, the expression of TIMP-1 in lesions of diffuse cutaneous systemic sclerosis and limited cutaneous systemic sclerosis were significantly higher than that of normal skins (both P<0.05). It was suggested that the expression of MMP-9 and TIMP-1 might play an important role in the development of systemic sclerosis.

CD46 is not only identified as a complement regulatory protein which protects host cells from complement attack, but also a new co-stimulatory molecule for human T cells. CD3/CD46 co-stimulation can induce a T-regulatory 1 cell (Tr1)-specific cytokine phenotype in human CD4(+) T cells. However, the role of CD46 as a co-stimulatory molecule in the modulation of the acquired immunity, such as transplant immunology, remains unclear. In this study, CD4(+) T cells were isolated from human CD46-transgenic C57BL/6 mice by magnetic-activated cell sorting, and further induced by anti-CD3, anti-CD28 and anti-CD46 antibodies respectively, and anti-CD3/anti-CD28 antibodies, anti-CD3/anti-CD46 antibodies, or the monoclonal antibody panel against CD3/CD28/CD46. The levels of interleukin-2 (IL-2), gamma-interferon (gamma-IFN), interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta) were detected in the supernatants of different groups. Suppression of allogeneic T cell proliferation were assessed by using mixed lymphocyte reaction (MLR) assay, in which monoclonal antibodies against CD46 were added to the culture. The results showed that CD3/CD28, CD3/CD46 and CD3/CD28/CD46 co-stimulation could significantly induce stronger proliferation of T cells than CD3 stimulation (P<0.05), and CD3/CD28/CD46 co-stimulation significantly increased the proliferation of T cells when compared with CD3/CD28 or CD3/CD46 co-stimulation (P<0.05 for each). IL-2 and gamma-IFN levels were much higher in CD3/CD28 co-stimulation group than in CD3, CD28, CD46 and CD3/CD46 groups (P<0.05 for each). IL-10 and TGF-beta levels were dramatically increased in CD3/CD46 co-stimulation group as compared with those in the CD3, CD28, CD46 and CD3/CD28 groups (P<0.05 for each). CD3/CD46 co-stimulation significantly inhibited the T cell proliferation and allogenic immune responses through the secretion of IL-10 and TGF-beta in MLR (P<0.05). These results suggested that CD3/CD46 can induce Tr1 cells to modulate allogenic immune responses, and it may become a novel target for the development of new therapeutic approach for T-cell-mediated diseases. CD46 plays an important role in regulating the T cell-mediated immune responses by bridging innate and acquired immunity.

AIM: To establish a sensitive and specific liquid chromatobraphy-mass spectrometry(time-of-flight)[LC-MS(TOF)] for the determination of astragaloside Ⅳ in rabbit plasma. METHODS: The HPLC-MS utilizing solid phase extraction was established to determine the concentration of astragaloside IV and ginsenoside R_~g1 , was used as internal standard. The analysis was carried on Agilent Hypersiol ODS(5 ?m, 4.6 mm?250 mm) column with a mobile phase of methanol-water (80∶20, v/v).Detection was performed on a time-of-flight mass spectrometry equipped with an ESI internal and operated in positive-ionization mode. Astragaloside Ⅳ quantitation was realized by computing the peak area ratio (astragaloside Ⅳ-ginsenoside R_~g1 )(astragaloside Ⅳ m/z807[M+Na]+ and ginsenoside R_~g1 m/z823[M+Na]+) and comparing them with calibration curve (r=~0.999 ). RESULTS: The linear calibration curve was obtained in the concentration range of 0.01-5 ?g?L~-1 .The detection limit of astragaloside Ⅳ was 0.01 ?g?L~-1 .The average recovery was more than 98%.The intra-and inter-run precision was measured to be below 5% of RSD. CONCLUSION: The method is sensitive, simple and rapid ,so, it can meet the need for the pharmacokinetics and bioavailability of astragaloside Ⅳ.

Cold Temperature ; Dinoprost ; analogs & derivatives ; urine ; Dust ; Humans ; Interleukin-6 ; urine ; Thromboxane B2 ; analogs & derivatives ; urine ; Weather

The molecular mechanism underlying muscular atrophy and gravisensing during spaceflight is still unknown. The major effects of spaceflight on body-wall muscles of Caenorhabditis elegans (C. elegans) in the structures and functions wore examined, and five important muscle-related genes and three proteins were studied after nearly 15-day spaceflight. The changes for the wall-muscles were observed in situ. Decreased muscle fiber size was observed with myosin immunofluorescence and duller dense-body staining in flight samples, which suggested that muscular atrophy had happened during spaceflight. However, F-actin staining showed no differences between the spaceflight group and ground control group. Otherwise, after returning to the earth the C eleganu displayed reduced rate of movement with a lower ratio (height/width) in crawl trace wave, which indicated a functional defect. These results demonstrated that C. elegans muscular development was changed in response to microgravity, and changes also occurred at the level of gene transcription and protein translation. Expression of dys-I increased significantly in body-wall muscles, while hlh-1, myo-3, uric-54 and eg1-19 RNA levels decreased after spaceflight. Dystrophin (encoded by dys-1) is one of important components in dystrophin-glycoprotein complex (DGC). Increased dys-I expression after flight implied that the muscular cell would accept more gravity signals by DGC in mierogravity in order to keep mechanical balance within the cells. It is concluded that DGC was involved into the mechanical transduction in body-wall muscles of C. elegans when gravity varied, which potentially played a vital role in gravisensing. The changes ofhlh-l, myo-3, tmc-54 and egl-19 suggested that they had the effects of promoting microgravity-induced muscular atrophy in strcture and function aspects. Result of Western blotting showed that the level of myosin A in spaceflight group decreased, further confirmed that atrophy happened during flight.

Objective To investigate the expression of caspase-10 in differentiated thyroid carcinoma and association with its development and metastasis. Methods Thyroid samples from 37 patients in a period from January 2006 to December 2007, with differentiated thyroid carcinoma were retrospectively analyzed for caspase-10 by immunohistocbemistry(streptavidin-perosidase, S-P), compared to control group of 46 cases with nodtdar goiter. The relationship between the expression of caspase-10 and the clinical pathologic characteristics of thyroid carcinoma were also explored simultaneously. Results caspase-10 were observed as brown or yellow particles located in the cytoplasm or cell membrane of nodular goiter but there were no significant evidence for its positive expression in thyroid carcinoma, caspase-10 expression was markedly down-regulated in differentiated thyroid carcinoma(29.73%,11/37) compared with benign nodules(71.74%,33/46, χ2=14.528, P<0.01). The positive expression in 18 cases with lymph node metastasis(11.11%,2/18) was significantly lower than those in 19 patients without lymph node metastasis(47.37%,9/19; χ2=4.210, P<0.01). There was no significant correlation(P> 0.05) between the expression of caspase-10 and the clinical pathologic characteristics including male, age, TNM stage and pathologic type. Conclusion Down-regulation of caspase-10 may play a critical role in carcinogenesis and development of differentiated thyroid carcinoma.

OBJECTIVETo present a method of quantitative diagnosis of craniofacial skeleton deformities based on three-dimensional computed tomography (3D CT).

METHODS20 cases with facial asymmetric deformities underwent 3D CT and the 3D images were reconstructed by Mimics 10.0 (Belgium). Anatomical landmarks were located and the coordinate of the landmarks obtained. Axial images of 1 patient with Romberg disease was used as representative case. The differences in the distance between the right landmarks and the left were calculated and analyzed.

RESULTSThe measurement results were not significantly different between two stages with an interval of 4 weeks ( P > 0.05), showing a reproducible resutls. The deviation of landmarks at facial midline increased gradually from upward to downward, reaching (2.63 +/- 0.54) mm at menton point. Paired landmarks showed asymmetry in three dimensions, especially gonion point on the left side, which was deviated 10.21 mm inward, 9.26 mm forward, 6.30 mm upward, compared to the opposite side.

CONCLUSIONSThe method of 3D CT quantitative analysis can provide precise information in the diagnosis and treatment planning of facial asymmetry deformity.

Anatomic Landmarks ; diagnostic imaging ; Cephalometry ; Craniofacial Abnormalities ; diagnostic imaging ; Facial Asymmetry ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; methods ; Tomography, X-Ray Computed ; methods

Objective To explore the treatment of cervical tracheoesophageal fistula (TEF) with complicated or remnant iaryngotracheal stenosis (LTS) and anterior neck defect (AND). Methods From 1980 to 2007,14 patients were diagnosed as TEF. Among them, 9 patients had complicated or remnant LTS, 3 patients had complicated AND, and 2 patients had TEF which were induced by Niekel-Titaium alloy mesh stent for treating benign esophageal stricture. All these patients were retrospectively studied in Tangdu Hospital. Treatment consisted of conservative therapy of TEF, staged surgical repair of TEF and laryngotracheal reconstruction according to the dimension (small or large) of TEF and complications. Results Four patients with small TEE (2-3 mm length) complicated LTS underwent laryngotracheal reconstruction stented with silicone T tube and TEF was adopted conservative treatment. The TEF and LTS were healed. Six patients with larger TEF (10-25 mm length) were repaired by staged surgical repair of TEF and laryngotracheal reconstruction. Among them, 3 eases had complicated LTS and AND, 2 cases had rement LTS and 1 ease had TEF without complication. Two patients had TEF and LTS, whose TEF healed before laryngntracheal reconstruction, the remnant LTS were reconstructed and healed. During the follow-up ranged from one to ten years, 12 patients were successfully treated without complications. One patient with TEF and LTS was treated only LTS because of a segment of esophagus was closed and treated with esophugngastrostomy in the department of thoracic surgery after LTS was successfully reconstructed and cured. One patient died of bleeding and asphyxia induced by the Nickel-Titanium alloy stentt because of the stent had not been taken out. Conclusion The small cervical TEF complicated or remnant LTS can be treated by laryngotracheal reconstruction and conservative treatment of TEF at the same time. A larger TEF complicated LTS should be treated by staged repair of TEF and LTS.

OBJECTIVETo sum up the clinical experience in the reparation of burn wounds with different types of skin flaps in patients with severe electrical injury.

METHODSFree skin flap, axile island flap with vascular pedicle and muscular skin flap were employed to repair 64 wounds in 49 cases.

RESULTSSkin flap necrosis happened in 2 cases, congestion and necrosis at the edge of skin flaps in 4 cases, and complete survival of the skin flaps and primary healing of the wounds in the remaining cases.

CONCLUSIONReparation of wounds with proper skin flaps according to the degree of the electrical injury is effective and reliable, which might be beneficial to the prevention of necrobiosis and the infection in deep tissue.

Adolescent ; Adult ; Burns, Electric ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps