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Wang J.,General Hospital of PLA | Sun L.,General Hospital of PLA | Si Y.-F.,309th Hospital of PLA | Li B.-M.,General Hospital of PLA
Molecular and Cellular Biochemistry | Year: 2012

The aim of present work was to elucidate the role of actin-depolymerizing factor (ADF), an important regulator of actin cytoskeleton, in the oxidized low-density lipoprotein (ox-LDL)-induced blood-brain barrier (BBB) disruption. The primary mouse brain microvascular endothelial cells (MBMECs) were exposed to ox-LDL. Treatment with LDL served as control. It was found that ADF mRNA level and protein expression were decreased when exposed to ox-LDL in MBMECs. Then, we investigated the influence of ADF overexpression on ox-LDL-treated MBMECs. Structurally, overexpression of ADF inhibited ox-LDL-induced F-actin formation. Functionally, overexpression of ADF attenuated ox-LDL-induced disruption of endothelial barrier marked by restoration of transendothelial electrical resistance, permeability of Evans Blue and expression of tight junction-associated proteins including ZO-1 and occludin, and blocked ox-LDL-induced oxidative stress marked by inhibition of reactive oxygen species (ROS) formation and activity of NADPH oxidase and Nox2 expression. However, overexpression of ADF in control cells had no significant effect on endothelial permeability and ROS formation. In conclusion, overexpression of ADF blocks ox-LDL-induced disruption of endothelial barrier. In addition, siRNA-mediated downregulation of ADF expression aggravated ox-LDL-induced disruption of endothelial barrier and ROS formation. These findings identify ADF as a key signaling molecule in the regulation of BBB integrity and suggest that ADF might be used as a target to modulate diseases accompanied by ox-LDL-induced BBB compromise. © 2012 Springer Science+Business Media, LLC.


An H.R.,309th Hospital of PLA
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] | Year: 2011

To study the possible relationship between polymorphic N-acetyltransferase 2 (NAT2) acetylator status and antituberculosis drug-induced hepatotoxicity and to elucidate the molecular mechanism of antituberculosis drug-induced hepatotoxicity. Blood samples from 101 tuberculosis cases with antituberculosis drug-induced hepatotoxicity and from 107 tuberculosis without antituberculosis drug-induced hepatotoxicity were collected for a case-control study. DNA of the subjects was extracted and amplified by polymerase chain reaction (PCR). The single nucleotide polymorphisms of NAT2 were determined by direct PCR sequencing. The genotype frequencies were compared between cases and controls by χ(2) test, using SPSS 12.0 software, and the association between the disease and genotypes was analyzed. Among the 101 patients with antituberculosis drug-induced liver injury, 36 patients (35.6%) were found with 282 T/T, 12 (11.9%) with 590 A/A, and 48 (47.5%) with 857 G/A or A/A. However, among the 107 controls, 9 patients (8.4%) were found with 282 T/T, 3 (2.8%) with 590 A/A, and 33 (33.8%) with 857 G/A or A/A. The patients with 282 T/T, 590 A/A, or 857 G/A or A/A genotype had a higher risk of antituberculosis drug-induced hepatotoxicity than those with 282 C/C or C/T, 590 G/G or G/A, or 857 G/G, and the OR values were 6.03 (95%CI: 2.88 - 12.62; χ(2) = 22.73, P < 0.05), 4.67 (95%CI: 1.42 - 15.44; χ(2) = 6.40, P < 0.05) and 2.03 (95%CI: 1.16 - 3.57; χ(2) = 6.08, P < 0.05) respectively. There were 40 patients with slow acetylator (39.6%) in cases with hepatotoxicity and 13 with slow acetylator (12.2%) in controls without hepatotoxicity. Patients with slow acetylator genotype (OR = 4.74, 95% CI = 2.42 - 9.28; χ(2) = 20.62, P < 0.05) had a significantly higher risk of antituberculosis drug-induced hepatotoxicity than those with rapid or intermediate acetylator genotypes. Among the cases, 19.8% (20/101) were found with NAT2(*)6A/7B, and 11.9% (12/101) with NAT2(*)6A/6A, whereas among the controls, 2.8% (3/107) were found with NAT2(*)6A/7B, and 2.8% (3/107) with NAT2(*)6A/6A respectively, the patients with NAT2(*)6A/7B and NAT2(*)6A/6A had a much higher risk of antituberculosis drug-induced hepatotoxicity, and the OR values were 8.40 (95%CI: 2.85 - 24.73; χ(2) = 14.90, P < 0.05) and 4.67 (95%CI: 1.42 - 15.44; χ(2) = 6.40, P < 0.05) respectively. Perhaps, the slow acetylation genotypes of NAT2 were the main risk factors of developing antituberculosis drug-induced hepatotoxicity.


Hu Y.L.,309th Hospital of PLA
Zhonghua zheng xing wai ke za zhi = Zhonghua zhengxing waike zazhi = Chinese journal of plastic surgery | Year: 2011

To examine the effect of RhoA/Rho kinase signal pathway on TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts. The 4th generation of primary cultured human dermal fibroblasts were stimulated with TGF-beta1, (10 ng/ml). The expression of alpha-SMA was detected after treatment with TGF-beta1, for 0, 3, 6, and 24 h. The expression of alpha-SMA was also detected after treatment with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml). Then the human dermal fibroblasts (4th generation) were stimulated with TGF-beta1, (10 ng/ml) after being treated with the RhoA/Rho kinase signaling pathway inhibitor Y-27632 (10 umol/ml). The fibroblasts were treated with nothing as sham control, or with Y-27632 (10 umol/L) only as negative control group, or with TGF-beta1 (10 ng/ml) only as positive control group. The expression of alpha-SMA was detected in all the groups. Protein expression was analyzed with ANOVA statistical method. alpha-SMA expression in fibroblasts with 10 ng/ml TGF-beta1 stimulation for 0, 3, 6, 24 h was 1.0, 1.9 0.2, 2.1 +/- 0. 1, 3. 1 +/- 0.1, respectively. Alpha-SMA expression in 24 h group was significantly higher than that in other three groups (n = 4, P < 0.05). alpha-SMA expression in human dermal fibroblasts after stimulation with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml) was 1.0, 1.4 +/- 0.2, 3.2 + 0.1, 3.1 +/- 0.2, respectively. alpha-SMA expression in 10 ng/ ml group was significantly higher than that in 2 ng/ml group and control group (n = 4, P < 0.05). There was no statistical difference in alpha-SMA expression between 10 ng/ml group and 50 ng/ml group (n = 4, P > 0.05). With both Y-27632 (10 micromol/L) and TGF-beta1 stimulation, the cell phenotype differentiation was inhibited. Alpha-SMA expression in experimental group (1.2 +/- 0.2) was significantly reduced, when compared with that in positive control group (2.9 +/- 0.1) (n = 5, P < 0.05). There was no significant difference (n = 5, P > 0.05) in alpha-SMA expression between control group (1.0) and negative control group (1.1 +/- 0.1). RhoA/Rho kinase signaling pathway should be involved in TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.


Chen Y.P.,309th Hospital of PLA
Zhonghua nei ke za zhi | Year: 2013

To evaluate the efficacy and safety of tacrolimus in patients with generalized myasthenia gravis (MG). A total of 69 cases admitted to our hospital were given 2-6 mg/day tacrolimus (FK506) for 12 months. The MG absolute and relative clinical scores were used to monitor the efficacy of tacrolimus. Clinical evaluation was conducted at month 1, 3, 6, and 12, while the serum concentration of FK506 was measured at one month after administration of tacrolimus for one month. The therapeutic response presenting as improved muscular strength showed within one month after administration of tacrolimus. The overall response rates (MG relative clinical score ≥ 25%) at month 1, 3, 6 were 81.2%, 87.6%, 92.2% respectively. It reached 93.8% by the final visit at month 12. MG score to evaluate disease severity decreased significantly as the subjects continued to take tacrolimus. Statistic analysis suggested that the serum concentration of FK506 was correlated with its therapeutic effect. Serum trough levels in remission and response groups [(7.1 ± 3.9) μg/L and (6.3 ± 3.8) μg/L, respectively] were significantly higher than that of no response group [(3.4 ± 1.3) μg/L]. The most common adverse effects included hyperglycemia (5 cases), myelosuppression (3 cases), and dizziness tinnitus (3 cases), majority of which were temporary and manageable. Our study has shown that tacrolimus significantly improved muscular strength of generalized MG patients. The treatment is well tolerated. The therapeutic effect of tacrolimus is observed within 1 month after initial use. Adverse events were manageable and not common.


Wang W.,309th Hospital of PLA
Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal of tuberculosis and respiratory diseases | Year: 2010

To describe the manifestations and diagnosis of pleural cavity extraskeletal osteosarcoma (ESO). One case of ESO diagnosed at the Research Institute of Tuberculosis, 309th Hospital of PLA was reported. Six cases reported in the literature were reviewed. Chest CT of a middle-aged man revealed an enormous heterogeneous neoplasm, about 10.9 cm x 9.2 cm x 17.7 cm in size, in the left pleural cavity. There was abundant calcification in the tumor, with signs of invasion into the diaphragm and the pleura. Pleural effusion of the left thoracic cavity was also seen on the chest CT. Osteosarcoma was confirmed by pathological study after surgical resection of the tumor. ESO is a rare malignant soft tissue sarcoma. Pleural cavity ESO is insidious and imaging studies often reveal a huge mass with abundant calcification. The differential diagnosis includes benign and malignant diseases of the thorax.


Chen Y.P.,309th Hospital of PLA
Zhonghua nei ke za zhi [Chinese journal of internal medicine] | Year: 2012

To investigation the clinical characteristics in myasthenia gravis (MG) patients with thymomas. A total of 856 MG patients admitted to the department during 2008.7 - 2010.12 were reviewed retrospectively. The patients with MG were divided into two groups based on thymic pathology, which were 162 cases with thymoma and 694 cases without thymoma. We compared the different clinical features including the gender, age of onset, MG symptoms and the incidence rate of myasthenia crisis. And the relationship between the WHO types, Maosaoka stages of thymoma and the severe of MG was also studied. The percentage of thymoma-associated MG patients was 18.9 percent of hospitalized MG patients at the same period. Of the 162 thymoma-associated patients, 94 were male and 68 were female, with a ratio of 1.38:1 and a mean age of (42.9 ± 12.4) years old. Thymoma was more frequent in middle-old aged patients than in children. Compared with non-thymoma MG, more thymomatous patients showed generalised MG, but not only ocular muscles weakness (90.1% vs 62.4%, P < 0.001). There were significant differences of the incidence rate of myasthenic crisis in the two groups (14.8% vs 2.3%). (2) WHO type B2 and Maosaoka I, II thymoma were the commonest types among all potentially MG-associated thymoma. No differences of Osserman MG classification was found in thymomatous patients with different pathologic changes. The thymomatous MG patients had its distinctive clinical features: thymomas occurred in about 19.7% of MG patients with more men than women, more common in generalized, higher incidence of myasthenia crisis, with B2 type thymic pathology and Maosaoka I, II stages. No correlation was found between pathologic and clinical stages.


WeiPeng,309th Hospital of PLA | Zhao G.,Chongqing Medical University | Ma Y.,309th Hospital of PLA | Yu H.,309th Hospital of PLA | Wang X.,Chongqing Medical University
Vaccine | Year: 2011

Recent evidence demonstrates that PEG10 plays an essential role in hepatocarcinogenesis and development, thus it could be regarded as a therapeutical target for hepatocellular carcinoma (HCC). In addition, transduction with recombinant, replication-defective adenoviral (Ad) vectors encoding tumor associated antigen into dendritic cells (DCs) is an efficient strategy to elicit antigen specific cytotoxic T lymphocytes (CTLs) for cancer therapy. In the present study, DCs were transduced with the PEG10 recombinant adenovirus, and were utilized to elicit CTLs in vitro.Moreover, the Trimera mice were immunized with the transduced DCs to elicit the immune response, the tumor growth and the life span of tumor bearing mice were observed. The results demonstrated that the transduced DCs could effectively induce specific CTL response against HCC without lysing autologous lymphocytes, but also significantly inhibit the tumor growth and prolong the life span of tumor bearing mice. These data suggest that PEG10 recombinant adenovirus transduced DCs can induce anti-tumor immunity against HCC expressing PEG10 in vitro and in vivo. Thus, the transduction of DCs with Ad-PEG10 provides a promising strategy for cancer immunotherapy of HCC. © 2011.


Loi P.,Free University of Colombia | Yuan Q.,Free University of Colombia | Yuan Q.,309th Hospital of PLA | Torres D.,Free University of Colombia | And 7 more authors.
Hepatology | Year: 2013

Interferon regulatory factor 3 (IRF3) is an important transcription factor in Toll-like receptor 4 (TLR4) signaling, a pathway that is known to play a critical role in liver ischemia-reperfusion injury. In order to decipher the involvement of IRF3 in this setting, we first compared the intensity of hepatic lesions in IRF3-deficient versus wildtype mice. We found increased levels of blood transaminases, enhanced liver necrosis, and more pronounced neutrophil infiltrates in IRF3-deficient mice. Neutrophil depletion by administration of anti-Ly6G monoclonal antibody indicated that neutrophils play a dominant role in the development of severe liver necrosis in IRF3-deficient mice. Quantification of cytokine genes expression revealed increased liver expression of interleukin (IL)-12/IL-23p40, IL-23p19 messenger RNA (mRNA), and IL-17A mRNA in IRF3-deficient versus wildtype (WT) mice, whereas IL-27p28 mRNA expression was diminished in the absence of IRF3. The increased IL-17 production in IRF3-deficient mice was functionally relevant, as IL-17 neutralization prevented the enhanced hepatocellular damages and liver inflammation in these animals. Evidence for enhanced production of IL-23 and decreased accumulation of IL-27 cytokine in M1 type macrophage from IRF3-deficient mice was also observed after treatment with lipopolysaccharide, a setting in which liver gamma-delta T cells and invariant natural killer T cells were found to be involved in IL-17A hyperproduction. Conclusion: IRF3-dependent events downstream of TLR4 control the IL-23/IL-17 axis in the liver and this regulatory role of IRF3 is relevant to liver ischemia-reperfusion injury. © 2012 American Association for the Study of Liver Diseases.


Ma Y.Z.,309th Hospital of PLA | Cui X.,309th Hospital of PLA | Li H.W.,309th Hospital of PLA | Chen X.,309th Hospital of PLA | And 2 more authors.
International Orthopaedics | Year: 2012

Purpose: There are few articles in the literature comparing outcomes between anterior and posterior instrumentation in the management of thoracic and lumbar spinal tuberculosis (TB). Methods: Between January 2004 and December 2009, 217 adult patients, average age 39 (range 16-67) years with thoracic and lumbar spinal TB were treated by anterior radical debridement and fusion plus instrumentation, anterior radical debridement with fusion and posterior fusion with instrumentation, posterolateral debridement and fusion plus posterior instrumentation or transpedicular debridement and posterior fusion with instrumentation in a single- or two-stage procedure. We followed up 165 patients for 22-72 (mean 37) months. Of these, 138 underwent more than three weeks chemotherapy with isoniazid, rifampin, pyrazinamide and ethambutol, and the remaining 27 underwent operation for neurological impairment within six to 18 hours of the same chemotherapy regimen. In no case did relapse occur. Apart from eight patients with skip lesions treated by hybrid anterior and posterior instrumentation, anterior instrumentation was used in 74 patients (group A) and 83 patients (group B) were fixed posteriorly. Results: In both groups, local symptoms were relieved significantly one to three weeks postoperatively; ten of 14 patients (71%) in group A and 14 of 19 (74%) in group B with neurological deficit had excellent or good clinical results (P>0.05). Erythrocyte sedimentation rates (ESR) returned from 43.6 mm/h and 42.7 mm/h, respectively, preoperatively to normal levels eight to 12 weeks postoperatively. Kyphosis degree was corrected by a mean of 11.5° in group A and 12.6° in group B, respectively (P<0.01). Correction loss was 6.8° in group A and 6.1° in group B at the last follow-up (P<0.01). Fusion rates of the grafting bone were 92.5% and 91.8%, respectively, at final follow-up (P>0.05). Severe complications did not occur. Conclusion: These results suggest that both anterior and posterior instrumentation attain good results for correction of the deformity and maintaining correction, foci clearance, spinal-cord decompression and pain relief in the treatment of thoracic and lumbar spinal TB providing that the opeartive indication is accurately identified. However, the posterior approach may be superior to anterior instrumentation to correct deformity and maintain that correction. © 2011 Springer-Verlag.


Guo L.X.,309th Hospital of PLA
Zhongguo gu shang = China journal of orthopaedics and traumatology | Year: 2010

OBJECTIVE: To investigate clinical outcome of short-course chemotherapy in retreating spinal tuberculosis after radical operation. METHODS: Forty-six retreating patients with spinal tuberculosis were included in this series, 29 males, 17 females with the age from 27 to 61 years (average of 43.7 years). All patients were treated with radical operation and short-course anti-tuberculous chemotherapy from March 2005 to March 2008. The tuberculous focus located thoracic spine in 17 cases, thoracic-lumbar in 13 and lumbosacral vertebrae in 16 cases. Of them, 5 cases had sinuses of tuberculosis and 7 cases had incomplete palsy in lower limbs (Frankel C-D). CT or MRI showed obvious sequestra, cold abscess within spinal focus. Surgical procedures including debridement, auto-bone grafting, and one-stage internal fixation, was performed at the 4 to 6 weeks after chemotherapy. Chemotherapy regimes were 3HRZ/6-9HRE in majority of patients. Clinical effect and focus healing were evaluated at follow-up period. RESULTS: Tuberculous symptoms and local pain of vertebral volume were obvious in all patients before chemotherapy,with average ESR 65.3 mm/h and average CRP 37.4 mg/L. After 4-6 weeks chemotherapy, tuberculosis symptoms and vertebral pain improved in all patients, and the average ESR decreased to 38.3 mm/1h, the average CRP decreased to 17.2 mg/L. Two to three months after operation, tuberculous symptoms and local pain relived in all patients,ESR and CRP became normal in 37 cases. Six to twelve months after operation, bonegraft complex in each patient became stable and there were no instrument loosening or deformity correction loss. Six patients with incomplete palsy recovered and 1 case improved from Frankel C to D grade. Focus healing was achieved in 44 cases (95.7%) after short-course chemotherapy (3HRZ/6-9HRE), and there were no resurgence in 2 to 4 years follow-up period. Drug fast 2 cases for RFP+INH cured at the 15 months after chemotherapy. CONCLUSIONS: Removed tubercular focus for the treatment of retreating spinal tuberculosis can improve clinical effect and shorten chemotherapy course.

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