Objective To establish the murine congenital infection model by MCMV and observe the pathological changes and infection status of brain tissue.Methods After anesthesia,mice who were pregnant 11-13.5 days (E11-13.5 d) were intra-amniotic injected one uterus by one with virus (MCMV K181 suspension,1 μl,1×103 PFU).The control group of the same period was intra-anmiotic injected with culture medium DMEM (1 μl).Carefully reset the uteruses and close the abdomen.After 5 days of separated feeding,kill the pregnant mice,take the fetus out of the uterus,anesthetize and kill them.Make frozen sections of these fetal brains.Some sections were stained using conventional HE method,to observe the pathological changes under the light microscope.Detect MCMV early antigen in the brain tissue by immunohistochemistry staining and immunofluorescence assay.Results The survival rates of the infected group were 71.9％.Compared with the control group,intra-amniotic inoculation of MCMV does not affect the rate of fetal survival,fetal absorption,fetal death and the average weight of the heads,but decrease their average weight of the bodies.The pathological changes are found in the brain tissue of the mouse in the infection group.Through enzyme immunohistochemistry assay,there are many MCMV infected cells in brain-ventricular zone,brain subependymal zone,cerebral cortex and hippocampus area in the infection group.Similar findings were observed by immunofluorescence method.Conclusion By intra-amniotic injection of MCMV suspension,murine model of MCMV congenital infection can be successfully established.This model could be used to study the mechanisms of encephalodysplasia caused by congenital CMV infection in vivo.

Objective To evaluate the value of CT-guided biopsy of deep-located lesion in the diagnosis of lymphoma. Methods CT-guided percutaneous biopsy was performed in 58 patients with 16-20 gauge core-needle biopsy. The locations of lesion involved mediastinun, lung, retroperitonurn, spleen, kidney, adrenalal and musculoskeletal system. Pathology examination included hematoxylin and eosin staining and immunohistochemical assays. Results In 56 out of 58 cases,the biopsy findings could be confirmed by histologic examination including correctly diagnosed 47 malignant lymphomas, corresponding to a sensitivity of 81%;with fuithecmace subclassification 42 of the 47(89.4%) could be as diagnosed malignant lymphomas on the basis of CT-guided biopsy. Conclusions Biopsy of deeply located lymphoma mass under CT guidance has high diagnostic accuracy and low complication rate with convenience for subclassification of malignant lymphomas.

Objective To study X-ray, CT and MRI features of the spinal giant cell tumors (GCT)and to assess the clinical applied value.Methods Thirty cases of GCT of spine (13 males and 17 females with ages ranging from 17 to 69 years) were reviewed. Allcases underwent radiography,CT was done in 22 patients and MRI was performed in 16 cases.Results One lesion localized in cervical spine,10 in thoracic, 6 in lumbar and another 13 lesions in the sacrum.Osteolytic destruction and vertebral compression were seen on X-ray film. The main CT signs were expanding bone destruction and soft tissue mass. MRI showed low to intermediate signal intensity on T_1WI while high signal on T_2WI.Conclusion X-ray, CT and MRI are of significant value in diagnosis of the spinal GCT, and play an important role in surgical planning.

Objective To compare the clinical utility of CT-guided percutaneous biopsies for bone destruction. Methods The retrospective analysis of pathologic outcomes of 89 cases guided by X-ray and MRI, were obtained by needle aspiration (n=13) of 18-20G tru-cut biopsy needles (n=22) and 11-13G Ostycut biopsy needles (n=40). Results Seventy five (84.3%) patients with percutaneous biopsy outcome with concordant results from specimens subsequently obtained at surgery, 14 patients (15.7%) showed pseudo-negative results but no pseudo-positive cases. No obvious differences in pathological results were obtained among these three methods. Conclusions ① CT-guided percutaneous biopsy is effective in the evaluation of skeletal destructive lesions; ② Appropriate selection of percutaneous biopsy method for different kinds of lesion could raise the diagnostie accurracy.

Objective To evaluate the accuracy of bispectral index (BIS)-guided closed-loop target controlled infusion (TCI) system in comparison with opened-loop manual TCI during anesthesia of biliary tract and pancreas surgeries.Methods Forty adult patients undergoing open surgery of biliary tract or pancreas under total intravenous anesthesia, including 17 males and 23 females, aged 18-75 years, falling into ASA physical status Ⅱ or Ⅲ, were randomly allocated into closed-loop group (group C, n=20) and opened-loop manual group (group M, n=20).In group M, the propofol effect-site concentration was adapted at the discretion of the anesthesiologist to reach and maintain a BIS as close as possible to 42-52.In the closed-loop TCI group, propofol was administered using the closed-loop anesthesia delivery system to reach and maintain atarget BIS of 42-52.The BIS values would be recorded automatically by the system at each second after it began to run.The anesthesia duration, unconsciousness time, endotracheal intubation time, recovery time and endotracheal extubation time were recorded.The total usage of propfol and remifentanil were calculated.The incidence rates of emergence agitation, postoperative nausea and vomiting and intraoperative awareness were recorded.The frequencies of vasoactive drug were recorded.MDAPE, Wobble, GS through BIS values were calculated.Results BIS was maintained within ±10% of target (excellent) for significantly longer time in group C (52.1±10.5)% than that in group M (37.6±5.8)% (P<0.05).BIS was maintained within ±(10%-20%) of target (good) for the same time in both groups.MDAPE in group C (10.1±2.2)% were significantly lower than those in group M (15.3±6.4)% (P<0.05).GS in group C (23.1±8.9)% was significantly lower than that in group M (33.5±15.8)%.The usages of propofol in group C ·kg-1·min-1 were similar to those in group M (0.12±0.03) mg·kg-1·min-1, and the usages of remifentanil in group C (0.12±0.03) μg·kg-1·min-1 were similar to those in group M (0.15±0.05) μg·kg-1·min-1.The frequencies of vasoactive drug were similar in both groups.There was one incidence of emergence agitation in groups M.Postoperative nausea and vomiting and intraoperative awareness didn't occur in both groups.Conclusion The depth of the anesthesia is maitained more appropriately and stable in the closed-loop group than that in manual administration group.

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.

Objective To observe effects of Kangshuai Yizhi Capsule on ATP in brain tissue and IL-6, TNF-α in serum of aging model rats, and explore the protective effects of the capsule on brain tissue.Methods Totally 72 rats were randomly divided into a normal group and a model group. The subacutely aging model rats were made by injectingD-gal, then aging rats were numbered and grouped by random number table into the model group, Kangshuai Yizhi high-dose group, Kangshuai Yizhi Capsule low-dose group and Naofukang group. All dose groups were received gavage by giving corresponding doses, while normal group and model group were given the same amount of saline everyday. After treated for 60 days, the activity of Na+-K+-ATP and Ca2+-Mg2+-ATP in brain tissue, and IL-6, TNF-α in serum were detected.Results Compared with normal group, Na+-K+-ATP and Ca2+-Mg2+-ATP were less active (P<0.05), but levels of IL-6 and TNF-α in model group were significantly higher, with statistical significance (P<0.05). Compared with model group, after treated with Kangshuai Yizhi Capsule, Na+-K+-ATP and Ca2+-Mg2+-ATP were more active, and IL-6 and TNF-α levels were down-regulated significantly in dose groups, with statistical significance (P<0.05). Meanwhile, Kangshuai Yizhi Capsule high-dose group showed the most obvious effect among dose groups.ConclusionKangshuai Yizhi Capsule has effects of enhancing activity of ATP in brain tissue and reducing level of proinflammatory factors.

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

BACKGROUND:The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.
OBJECTIVE:To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.
METHODS:THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups:mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. cells in each group were col ected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups:HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cellnumber was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.
RESULTS AND CONCLUSION:After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly;compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100%2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

Objective To explore the imaging manifestations of low-grade central osteosarcoma (LGCOS) and discuss their pathological features.Methods Twelve patients of LGCOS proved by surgery and pathology were analyzed retrospectively and a review of related literature was performed.All twelve patients had plain X-ray,1 1 patients CT examination,and 10 patients contrast-enhanced MR scan.Imaging features of the LGCOS were summarized,their clinical and pathological manifestations were discussed for differential diagnosis.Their prognosis was evaluated with followed up examination.Results Of the 12patients with LGCOS,six tumors were located in the distal femur,3 in the proximal tibia,2 in the proximal femur and 1 in the talus.The radiographic features of LGCOS were variable.There were 7 patients with predominantly osteolytic destruction,3 patients with mixed sclerotic and lyric changes,with well-defined margins,2 patients with Osteogenic changes on X-ray.On CT,9 patients showed a clear cortical breach,5 patients with soft tissue involvement,6 patients with peripheral incompletely sclerotic zone,2 patients with periosteal reaction.On MRI,there were 10 patients with abnormal signal in medullary cavity,8 patients with soft tissue masses,and all 10 patients exhibited contrast enhancement.The microscopic features of LGCOS were characteristically bland,comprising spindle cells arranged in interlacing fascicles in a heavily collagenous background with variable bone or osteoid production.There were mild nuclear atypia and rare mitoses.In four patients,misdiagnoses were made by biopsy or surgical pathology as fibrous dysplasia or fibrous histiocytoma and other benign lesions,all four patients had postoperative recurrence.Conclusions LGCOS should be differentiated form fibrous dysplasia,non-ossifying fibroma,and other benign lesions.An accurate diagnosis can be made in most cases by careful pathological and radiological correlation.