BACKGROUND: Fat embolism syndrome is a commonly seen severe complication in the field of opthopaedics in clinic. It frequently occurs after long bone fracture and hip-knee joint replacement. However, its etiological factors and pathogenesis are not identified.OBJECTIVE: To sum up the onset influencing factors, pathogenesis and therapeutic methods of long bone fracture and hip-knee joint replacement accompanied with fat embolism syndrome.RETRIEVE STRATEGY: Using the terms "fat embolism syndrome, pathogenesis, treatment, prevention, diagnosis", we retrieved PubMed database to identify studies published in the English language. Fifty-five literatures were searched.Meanwhile, we searched the medical information network of Shanghai Jiao Tong University with the same terms in the Chinese language. Inclusive criteria: studies, which can reflect the diagnosis and treatment as well as pathogenesis of fat embolism syndrome. Exclusive criteria: repetitive studies.LITERATURE EVALUATION: The involved 38 literatures are all about the diagnosis and treatment as well as pathogenesis of fat embolism syndrome, among which, 6 were review and the others were clinical or basic studies.DATA SYNTHESIS: ① The influencing factors of fat embolism syndrome included trauma factor, operation factor and other factors. ② The pathogenesis of fat embolism syndrome involved mechanical obstruction theory, biochemical theory,condensation theory and inflammatory reaction theory. ③ The treatments of fat embolism syndrome include respiration supporting, glucocorticoid application, protecting brain function, drug treatment, heat shock treatment and so on.CONCLUSION: Study on bone fracture and joint replacement accompanied with fat embolism syndrome can provide evidence for the diagnosis and treatment of this syndrome.

Objective To observe expression and significance of inflammatory factor during perioperative and follow-up periods after endovascular stents in patients with cerebral arterial stenosis.Methods 54 patients diagnosed as cerebral arterial stenosis by digital substraction angiography (DSA) were selected as the stent group and treated with endovascular stents; another 32 subjects had the same disease but not accepted stenting were considered as the control group. Interleukin-6 (IL-6) and high-sensitivity C-reactive protein (hs-CRP) contents were measured at different time points during perioperative and follow-up periods in the two groups. Stage A represented as day one before angiography or catheterization; stage B as 6 hours postoperatively (stent group) or 6 hours after diagnostic angiography (control group); stages C～I as 12, 24, 48, 72 hours, 1 and 6 months after stent insertion (stent group) or the same time points after angiography (control group).Results Contents of IL-6 and hs-CRP of the stent group were similar as the control group in the stage A ( P>0.05), but significantly higher than that of the control group during stages B～I ( P<0.01～0.05). Among 54 patients of the stent group, 21 cases had restenosis 6 months postoperatively (38.89%). Contents of IL-6 and hs-CRP of the patients were similar as those without restenosis in stages A～F postoperatively ( P>0.05), but significantly higher than that of the cases without restenosis in stages G and I postoperatively ( P<0.01～0.05).Conclusion Endovascular stents can increase the contents of IL-6 and hs-CRP of patients with cerebral arterial stenosis; in addition, high expression of them is the risk factor of post-stent restenosis during follow-up period.

ObjectiveTo determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of granulocyte colony-stimulating factor (G-CSF), and the role of Toll-like receptor 4 (TLR4) signaling pathway in this process.Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. ExperimentⅠ: the cells were divided into four groups as follows: control group, LPS stimulation group (LPS 10μg/mL), LPS+ 0.1 U/mL UFH group, and LPS+ 1 U/mL UFH group. HPMECs in UFH groups were treated with 0.1 U/mL or 1 U/mL UFH 15 minutes before LPS stimulation, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The concentrations of interleukin-6 (IL-6) and G-CSF in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 24 hours after LPS challenge to detect the effect of UFH on HPMECs. ExperimentⅡ: HPMECs were treated with 5μg/mL of rhodobacter sphaeroides LPS (LPS-RS, antagonist for TLR4) 4 hours before the addition of PBS or LPS. The concentrations of IL-6 and G-CSF in cell culture supernatants were determined 24 hours after LPS stimulation to detect the effect of TLR4 on LPS-induced HPMEC injury. ExperimentⅢ: HPMECs were divided into four groups as before: control group, LPS stimulation group, LPS+ 0.1 U/mL UFH group, LPS+ 1 U/mL UFH group. Treatments to cells were the same as experimentⅠ. The protein expression of TLR4 in HPMECs was determined by Western Blot 1 hour after LPS stimulation to detect the effect of UFH on TLR4.Results① Compared with control group, the levels of IL-6 and G-CSF in LPS stimulation group were increased [IL-6 (ng/L): 655.9±58.3 vs. 75.5±18.2, G-CSF (ng/L): 388.7±36.2 vs. 35.3±12.6, both P< 0.05]. Compared with those of LPS stimulation group, in LPS+ 0.1 U/mL UFH group and LPS+ 1 U/mL UFH group, the levels of IL-6 and G-CSF were significantly decreased [IL-6 (ng/L): 518.2±64.6, 489.1±75.6 vs. 655.9±58.3, G-CSF (ng/L): 298.8±41.0, 273.4±33.2 vs. 388.7±36.2, allP< 0.05]. The results indicated that 1 U/mL UFH had better results, though there was no statistical significance between the results of two UFH groups.② LPS-induced up-regulation of IL-6 and G-CSF levels was prevented by LPS-RS [IL-6 (ng/L): 139.1±37.6 vs. 655.9±58.3, G-CSF (ng/L): 73.7±19.7 vs. 388.7±36.2, bothP< 0.05]. LPS-RS alone had no effect on cytokines [IL-6 (ng/L):118.2±42.1 vs. 75.5±18.2, G-CSF (ng/L): 48.4±26.8 vs. 35.3±12.6, bothP> 0.05].③ Compared with control group, the protein expression of TLR4 (grey value) in LPS stimulation group was significantly upregulated after 1 hour (0.87±0.23 vs. 0.36±0.12,P< 0.05). UFH with 0.1 U/mL and 1 U/mL lowered TLR-4 protein expression induced by LPS (0.68±0.18, 0.62±0.26 vs. 0.87±0.23, bothP< 0.05).ConclusionsThe expressions of IL-6 and G-CSF were increased obviously in LPS treated HPMECs. UFH might take its therapeutic effect through TLR4-dependent pathway.

Purpose To explore the role of cell blocks combined with immunohistochemical examination in the diagnosis and differential diagnosis of malignant pleural effusion,and to explore the role of pleural effusion cell blocks in lung adenocarcinoma molecular pathology examination.Methods 142 cases of malignant pleural effusion based cytology,cell blocks of HE staining and immunohistochemical staining by EnVision twostep were retrospectively analysed,the tumor classification was made through analyzing the characteristics of the cells combined with antibody expression.The detection of epidermal growth factor receptor (EGFR) gene mutation of 40 cases of lung adenocarcinoma diagnosed after immunohistochemical staining were used by ARMS-PCR method.Results Among 142 cases of malignant pleural effusion,there were 99 cases caused by lung adenocarcinoma,4 cases of lung small cell carcinoma,3 cases of lung squamous cell carcinoma,13 cases of breast carcinoma,9 cases of ovarian carcinoma,2 cases of gastric carcinoma,1 case of thyroid carcinoma,1 case of endometrial carcinoma,5 cases of mesothelioma,3 cases of lymphoma,1 case of malignant melanoma,1 case of synovial sarcoma.In 40 cases of lung adenocarcinoma pleural effusion cell block,there were 20 cases with EGFR mutations,9 cases of 19del mutations and 11 cases L858R mutations.Conclusion The pleural effusion cell blocks combined immunohistochemistry are useful to improve the accuracy of diagnosis of patients with pleural effusion,and helpful for the determination of classification and the primary site of tumor,assessment of prognosis.Pleural dffusion cell block may used to detect EGFR mutations of lung adenocarcinoma,which provide new source of specimen for the gene detection of lung adenocarcinoma.

Thyroid function ultimately depends on appropriate iodine supply to the gland. Thyroid hormone deiodination is an intrinsic component of the thyroid hormone homeostasis. Type Ⅰ iodothyronine deiodinase (D1) plays an important role in thyroid hormone metabolism and has close relationship with thyroid function. Based on successfully establishing animal models of iodine deficiency and iodine excess in Babl/c mice (Babl/c mice were randomly divided into five groups: low iodine (LI), normal iodine (NI), five-fold iodine (5HI) , ten-fold iodine (10HI) and fifty-fold iodine (50HI) group. Three months and six months after admistration, they were sacrificed and thyroids were excised), the expression level of D1 mRNA were examined by using real time quantitative PCR method. D1 activity was analyzed by 125I-rT3 as substrate combined with ion-exchange chromatography. The thyroid hormone was measured with radioimmunoassay method. The data revealed that in the case of iodine deficiency, both D1 mRNA expression and D1 activity was greatly increased(compared with NI groups, P

AIM:To observe the effects of prostaglandin E2 receptor1 (EP1) in neuronal cell death induced by hypoxia/reoxygenation and ischemia/reperfusion. METHODS:The cortical neurocytes of neonatal Wistar rats were cultured for 12 days and exposed to hypoxia/re-oxygenation to establish a hypoxia/re-oxygenation model. Another set of cultured primary neonatal cortical neurocytes of rats were pretreated with 17-pt (an antagonist of EP1),then underwent hypoxia for 3 h,re-oxygenated for 21 h. MTT reagent was added 1 h before measuring the cell viability. Neuron apoptosis was determined by flow cytometry. The protein expression was examined by Western blotting. RESULTS:Compared to the control cells (only underwent hypoxia /re -oxygenation and without any pretreatment),the neurons pretreated with 17-pt and then underwent hypoxia/re-oxygenation showed significantly lower survival rate (P

Objective To investigate the binding protein of chlamydiaphage phiCPG1 capsid protein Vp1 on chlamydia trachomatis outer membrane.Methods The bacterium with recombinant plasmid Vp1/pet30a( + ) was induced.The expressed protein was purified by gel recycling.FarWestern blot was utilized to' investigate the binding protein of Vp1 on chlamydial outer membrane,including recombinant polymorphic outer membrane protein (rPmp) and major outer membrane protein (MOMP).Results The recombinant protein Vp1 was successfully expressed in E.coli.Monoclonal antibody against Vp1 was used as primary antibody in Western blot,and no specific band was present,which indicated that the monoclonal antibody did not specifically bind with any rPmp.Far-Western blot results showed that there was an obvious band for the rPmpI,but no specific band for other rPmp and MOMP,which suggested that Vp1 could specifically bind with rPmpI protein on the chlamydial outer membrane of serotype D.Conclusions There is a binding site of Vp1 on the chlamydia trachomatis outer membrane.Vp1 may play an important role in the interaction between the chlamydiaphage and the chlamydiae.

Aim To examine the inhibitory effect of re-combinant cardiac troponin fusion protein composed of subunit I and artificial peptide which was called CIS on tumor growth. Methods The CIS ’ s effect on the growth of human umbilical vein endothelial cells ( HU-VEC) was examined using MTT assay in vitro. Chick chorioallantoic membrane model was applied to study the alteration of angiogenesis treated with purified re-combinant CIS protein. The effect of tumor growth trea-ted with CIS was observed using several in vivo mice xenograft models. Results There was a statistically significant reduction in HUVEC cell proliferative rate when the cells were treated with purified CIS fusion protein, which was also shown in a dose-dependent manner. A decreased amount of new blood vessel for-mation ( angiogenesis) on chick embryo chorioallantoic membranes was observed in recombinant CIS protein treated group compared to the untreated control group. A significant inhibition of tumor growth rate was a-chieved in CIS treated mice compared to CIS untreated control mice in 6 different mouse xenograft models. Conclusions The fusion protein CIS shows the inhibi-tory effect on the tumor growth in our in vivo mouse models, and such inhibition could be mediated by the mechanism of CIS’ s effect on the decrease of HUVEC cell proliferation and further the reduction of angiogen-esis in tumor tissues. This work could provide the foundation for the in-depth investigations on the phar-maceutical application of CIS targeting anti-tumor ther-apy.

Objective To add an open reading frame in the shuttle vector of pGFP ∷ CM for transfection of exogenous genes into Chlamydia muridarum.Methods The sequence of plasmid pGFP ∷ CM and new open reading frame (including promoter of pgp4,mCherry gene of red fluorescence protein and transcription termination sequence of Chlamydia trachomatis CT579) were amplified by polymerase chain reaction (PCR),and the products were transfected into Stellar competent cells.The recombinant plasmids were identified by PCR,enzyme digestion and sequencing.Then the recombinant plasmid was transfected into plasmid-free strain CMUT3,and the GFP-and mCherry-positive inclusions were observed under the fluorescence microscope.After the ampicillin selection and plaque purification,the purified CMUT3-pGFP-mCherry-CM was identified by indirect immunofluorecesent stain using anti-pgp3 and anti-glgA antibodies.Results The correct recombinant plasmid after sequencing identification,enzyme digestion and PCR amplification was successfully transfected into CMUT3,and the GFP-and mCherry-positive inclusions were observed.The transfected strain CMUT3-pGFP-mCherry-CM was purified after ampicillin selection and plaque purification.The expression of pgp3 and glgA protein in CMUT3-pGFP-mCherry-CM was similar to that in CMUT3-pGFP ∷ CM.Conclusion An open reading frame is successfully added in the plasmid pGFP ∷ CM,and the new plasmid can be transfected into CMUT3 and express exogenous protein,which can be used for further study on the function of single chlamydial protein.

BACKGROUND: Evidence exists that inhibition of matrix metanoproteinase-2(MMP-2) secretion in the proliferating hernangioma tissue by transfection of adenovirus-active MMP-2(Ad-aMMP-2) cDNA would become an important means for treatment of proliferating hemangioma.OBJECTIVE: To investigate the influences of Ad-aMMP-2 cDNA transfection on human proliferating hemangioma growth in nude mice.DESIGN, TIME AND SETTING: A randomized, grouping, and controlled observation was performed in West China Hospital of Sichuan University between August 2003 and September 2004.MATERIALS: Eighteen BALB/c-nu/nu nude mice, weighing approximately 20 g, were included. Cavernous hemangioma specimen pathologically confirmed as proliferating hemangioma was resected from one 52-day-old female child patient.METHODS: The freshly reseoted human proliferating hemangioma specimen was sliced into small pieces with a size of 5 mm×4 mm×3 mm and subcutaneously implanted into the back of 18 nude mice within 1 hour to develop mouse models of hemangioma.Forty-five days after hemangioma implantation, 15 successful hemangioma nude mice were treated by intratumoral administration of adenovirus green fluorescent protein (Ad-GFP1 n = 51 Ad-GFP group), adenovirus-active MMP-2 (n = 5, Ad-aMMP-2 group), or the same amount of phosphate buffered saline (PBS1 n = 51 control group). Intratumoral administration was performed once every other day, for a total of 4 times.MAIN OUTCOME MEASURES: Observation of tumor volume and compadson of tumor necrosis area among 3 groups; detection of GFP expression in nude mouse; gross, hematoxylin-eosin staining, and transmission etectron microscope observation of tumor tissue morphology; determination of MMP-2 cDNA expression and microvascular density by immunohistochemistry; and detection of growth cycle and apoptosis of tumor cells by flow cytometry.RESULTS:①Ad-aMMP-2 could inhibit hemangioma growth in vivo, without marked adverse reactions. Tumor necrosis of different degrees was found in each group, and tumor necrosis area was significantly greater in the Ad-aMMP-2 group than in the control and Ad-GFP groups (P < 0.01). ②Histological sections displayed GFP gene expression in the Ad-GFP group. ③Gross observation results revealed relatively large tumor tissue in the control and Ad-GFP groups and relatively small tumor tissue in the Ad-aMMP-2 group. Hernatoxylin-eosin staining results showed that in the control and Ad-GFP groups, endothelial cells aggregated together in strip-shaped or lump-shaped appearance, and in the Ad-aMMP-2 group, there were many necrotic loci arranging in lamellar-shape appearance. Transmission electron microscope results revealed vascular endothelial cells with normal morphology in the control group and tumor cells with apparent nucleoli in the Ad-GFP group, while in the Ad-aMMP-2 group, some vascular endothelial cells exhibited chromatin pycnosis in the nucleus, forming apoptotic bodies.④ MMP-2 expression and microvascular density were significantly reduced in the Ad-aMMP-2 group than in the Ad-GFP and control groups (P < 0.05). ⑤The percentage of tumor cells in G0/G1 phase was significantly higher (P < 0.05), while the proliferating index was significantly decreased, in the Ad-aMMP-2 group than in the Ad-GFP and control groups. The Ad-aMMP-2 group exhibited higher apoptosis rate of tumor cells (P < 0.05), as well as more markedly increasing apoptosis index, than the control and Ad-GFP groups.CONCLUSION: It is feasible to block human proliferating hemangioma growth by transfeotion of Ad-aMMP-2 cDNA. The included mechanisms are to inhibit vascular endothelial cells to secrete MMP-21 thereby leading to local ischemia.