Daxing Zhu

Thymoma With Pure Red Cell Aplasia and Good's Syndrome

Thymoma patients with pure red cell aplasia (PRCA) or hypogammaglobulinemia (Goods syndrome) are rare, whereas those with both PRCA and Goods syndrome are even rarer. Here we present the case report of a 70-year-old woman with invasive thymoma and simultaneous PRCA and Goods syndrome, who achieved complete PRCA remission after thymectomy.

nm23-H1 is a negative regulator of TGF-β1-dependent induction of epithelial–mesenchymal transition

Members of transforming growth factor-β(TGF-β) family are the main inducers of epithelial-mesenchymal transition (EMT) during embryogenesis and cancer pathogenesis. However, a significant crosstalk between TGF-β and other signals occurs during the induction of EMT. nm23-H1 was the first metastasis suppressor gene to be identified on the basis of an inverse relationship between nm23-H1 expression and metastasis stage. Despite extensive studies, the mechanism underlying its ability to suppress metastasis is far from elucidated. We demonstrated here that the nm23-H1 negatively regulated TGF-β1-dependent induction of EMT in non-aggressive lung cancer cell line. nm23-H1 knockdown significantly enhanced TGF-β1-induced suppression of epithelial marker E-cadherin and upregulation of mesenchymal markers β-catenin and fibronectin. The invasive and migratory potential of lung cancer cells upon TGF-β1 treatment was also markedly enhanced by nm23-H1 knockdown. On the other hand, the effect of nm23-H1 depletion on TGF-β1-induced EMT was reversed by ectopic re-expression of shRNA-resistant nm23-H1 protein. Furthermore, TGF-β1-induced EMT potentiated by nm23-H1 depletion was partially dependent on transcriptional factor Snail expression. Finally, we found Src kinase is involved in regulation of TGF-β1-induced EMT by nm23-H1. Our results suggest a means of restoring nm23-H1 to suppress TGF-β1-induced EMT that may exploited therapeutically for the management of metastasis diseases.

Mediastinal solitary fibrous tumor with right diaphragm invasion: Report of a case

Mediastinal solitary fibrous tumors (SFTs) are rarely found in adults and there are few reports describing primary mediastinal SFT invading the diaphragm. We report the case of a 47-year-old woman with a large right inferior mediastinal SFT. Magnetic resonance imaging showed the tumor invading the right lower lobe of the lung and the right hemidiaphragm, with displacement of the inferior vena cava (IVC) and right lobe of the liver. Angiogram showed IVC stenosis. To our knowledge, this is the first report of complete resection of the tumor combined with right lower lobectomy of the lung and partial resection and reconstruction of the right diaphragm with a Dacron flap.

Metastasis suppressor Nm23-H1 inhibits STAT3 signaling via a negative feedback mechanism.

Persistent STAT3 activation is a critical event in tumorigenesis and metastatic progression. Recent studies have found higher levels of STAT3 in metastatic tissues than in primary tumor tissues. We speculated that such increased STAT3 activity might be attributed to a loss of function or reduction in expression of metastasis inhibitory protein during cancer progression, and we therefore examined the role of tumor metastasis-suppressor nm23-H1 in the activation of STAT3 in the A549 lung cancer cell line. We found that IL-6-dependent induction of tyrosine phosphorylation and activation of STAT3 were influenced by nm23-H1 inhibition. IL-6-induced STAT3(Tyr705) phosphorylation was significantly enhanced in A549 cells transfected with siRNA specific for nm23-H1, and the effect of nm23-H1 depletion on IL-6-induced STAT3(Tyr705) phosphorylation was reversed by ectopic expression of shRNA-resistant nm23-H1 protein. Moreover, STAT3 directly bound to the STAT3 binding site on the nm23-H1 promoter and activated its expression. Thus, we have identified a new feedback mechanism that might provide insight into an in-built metastasis-suppression function in tumor cells and which could be a logical new target for treatment of early metastatic disease.

[Lentivirus-mediated stable silencing of nm23-H1 gene in lung cancer cells and the influence on biological behavior].

背景与目的 nm23-H1基因是重要的肿瘤转移抑制基因。前期研究发现利用化学合成的小干扰RNA（small interfering RNA, siRNA）抑制nm23-H1基因的表达可明显增强肺癌细胞的侵袭力。为了进一步研究nm23-H1基因沉默后的分子生物学机制，本研究利用慢病毒介导的短发夹RNA（short hairpin RNA, shRNA）建立nm23-H1基因稳定沉默的肺癌细胞株。 方法 将表达特异性抑制nm23-H1基因shRNA的慢病毒转染人大细胞肺癌细胞株NL9980和肺腺癌细胞株A549，通过嘌呤霉素筛选出稳定转染细胞株。逆转录PCR、定量PCR及Western blot法检测nm23-H1基因表达，并通过shRNA抵抗的nm23-H1基因重组质粒转染拯救实验验证，侵袭小室实验检测侵袭力改变。 结果 逆转录PCR、定量PCR和Western blot法检测稳定转染细胞株NL9980-99和A549-99中nm23-H1基因在mRNA和蛋白水平表达均明显降低；shRNA抵抗的nm23-H1基因重组质粒转染拯救实验重现nm23-H1的正常表达；侵袭小室实验显示NL9980-99和A549-99细胞侵袭力明显增强。 结论 成功建立nm23-H1基因稳定沉默的人大细胞肺癌细胞株NL9980-99和人肺腺癌细胞株A549-99，nm23-H1基因沉默后使NL9980-99和A549-99细胞的侵袭力明显增强。

The N-terminal kinase suppressor of Ras complex has a weak nucleoside diphosphate kinase activity

Introduction:  An increasing number of studies have proven that the kinase suppressor of Ras (KSR1) functions as a scaffolding protein that coordinates the assembly of a multiprotein complex containing mitogen‐activated protein kinase and its upstream regulators. However, a few studies have reported that KSR1 can activate c‐Raf‐1. Therefore, whether KSR1 possesses a kinase activity has been an unresolved issue until now.

Bronchoplastic procedures and pulmonary artery reconstruction in the treatment of stage III lung cancer invading pulmonary artery

BACKGROUND To summarize the clinical results of bronchoplastic procedures and pulmonary artery reconstruction or combined with other resection and plasty of heart, great vessels in the treatment of 304 patients with locally advanced lung cancer. METHODS From February, 1983 to December, 2001, double sleeve resection and reconstruction of bronchus and pulmonary artery, or combined with other resection of heart, great vessels were carried out in 304 patients with locally advanced lung cancer. The operations included double sleeve left upper lobectomy in 199 cases; double sleeve right upper lobectomy in 21 cases; double sleeve right upper middle lobectomy in 14 cases; double sleeve left upper lobectomy combined with resection of left atrium in 8 cases; double sleeve right upper lobectomy combined with superior vena cava (SVC) resection and reconstruction with Gortex graft in 29 cases; double sleeve right upper middle lobectomy combined with SVC resection and reconstruction in 21 cases; double sleeve right upper middle lobectomy, carinal and SVC resection and reconstruction in 11 cases; left pneumonectomy combined right main pulmonary artery and pulmonary artery trunk resection and reconstruction with Gortex graft in 1 case. RESULTS There were 3 operative deaths. The operative mortality was 1% in this series. Sixty four patients had operative complications. The operative complication rate was 21.05% (64/304). The 1-, 3-, 5- and 10 year survival rates were 81.75%, 60.14%, 37.21% and 24.39% respectively. CONCLUSIONS Double sleeve lobectomy or comblined with other resection and reconstruction of heart, great vessels can significantly improve the prognosis and increase the curative rate and long term survival in patients with locally advanced lung cancer.

[Establishment of the cell line that human lung adenocarcinoma can stably express luciferase which is absent of nm23-H1 expression and detecting its luminescence in vitro and in vivo].

BACKGROUND AND OBJECTIVE On the condition that laboratory animals survive, we can detect the distribution of tumor cells by in vivo imaging that were labeled with firefly- luciferase (luc) gene. The purpose of this study is to establish a light-emitting cell line A549/nm23-H1-shRNA-luc which could express nm23-H1 shRNA and firefly-luciferase stably, and detect its bioluminescence in vitro and in vivo. It will provide the preparation for the next related experimental research in vivo. METHODS The optimal concentration of hygromycin B for screening A549/nm23-H1-shRNA cells was determined by concentration gradient method. We firstly transfected the plasmid (PGL4.50) with luc gene into A549/nm23-H1-shRNA cells and then screened the monoclonal cell line A549/nm23-H1-shRNA-luc with hyhromycin B. The positive monoclonal cell line was identified with an in vivo imaging system, thereafter the expression stability of luciferase was analyzed in the strongest light-emitting positive monoclonal cell line. The A549/nm23-H1-shRNA-luc cells were inoculated subcutaneously into right-hind groin of nude mice and then observed by the in vivo imaging system. RESULTS The optimal concentration of hygromycin B used in screening A549/nm23-H1-shRNA cells was 300 μg/mL. After screening, the A549/nm23-H1-shRNA-luc cells established can express luciferase stably in vitro, a great linear correlation existed between the amount of cells (x) and bioluminescence values (y), with an equation of y=3,699.9x+992,237, and the square of the correlation coefficient (R2) was 0.975,1. To evaluate the stability of bioluminescence in vivo, 10 nude mice were randomly divided into two groups that the same number of cells were implanted into. The variation of bioluminescence values detected in vivo between the two groups of the same cells was not statistically significant (P>0.05). CONCLUSIONS We have successfully established the cell line A549/nm23-H1-shRNA-luc which can express luciferase persistently and stably.

Giant Primary Chondroma of the Lung: A Case Report

患者,女,40岁,查体发现右下肺肿物1周入院。 既往体健。体格检查:体温正常,脉搏、血压、呼吸 均无异常。胸壁无压痛,双肺呼吸运动对称,触觉语颤 对称,叩诊呈清音,听诊双肺呼吸音稍粗,未闻及干、 湿啰音。入院后CT检查:右下肺后基底段可见大小约 6.4 cm×5.7 cm×7.5 cm的不规则软组织肿块,内可见多发 斑片钙化,形态不规则,肿块与局部胸膜粘连,未见明 显强化(图1)。完善术前检查后于2010年8月17日在全 麻下行剖胸探查,术中见包块位于右肺下叶后段,突出 于肺表面,不规则分叶,被覆脏层胸膜及少量肺组织, 与周围组织无粘连,质硬。行右肺下叶楔形切除术,完 整切除肿物。包块为灰白灰褐不规则组织,大小6.5 cm× 6.0 cm×5.0 cm,有多个结节状突起,切面质地较硬,灰 白色,半透明状,软骨样(图2)。镜检:瘤体由较成 熟的软骨细胞构成,周围为软骨基质包绕,呈不规则分 叶状(图3)。病理诊断:右下肺软骨瘤,局部生长活 跃。患者术后10天治愈出院。

[Construction, Expression and Purification of Wild and Mutant Type of nm23-H1 in Prokaryotic Expression System.].

BACKGROUND Nm23-H1 is a metastasis-suppressor gene. However, its molecular mechanism of suppressing metastasis is unknown until now. The aim of this study is to construct prokaryotic expression vector of wild and mutant type of nm23-H1 (WT, P96S, H118F), and then express and purify the proteins. METHODS wild and mutant type of nm23-H1 fragments were amplified by PCR. The prokaryotic expression vectors of pET28anm23-H1 were constructed by gene recombination technique and verified by restriction enzyme analysis and sequencing. The positive clones were transformed into E. coli BL21 (DE3) and soluble analysis of the expression was conducted in this system. The proteins were purified by nickel column chromatography and identified by Western blot RESULTS The sequences and open read frames of all the pET28a-nm23-H1 plasmids were completely correct. After transforming, these plasmids can express the target proteins. The protein production was very high, and all the proteins were soluble expression. The molecular weight of wild and mutant type of nm23-H1 was 20 kDa detected by Western blot, which was as the same as the objective protein. CONCLUSIONS We have succeeded in constructing the prokaryotic expression vectors of pET28a-nm23-H1 (WT, P96S, H118F) and the proteins which expressed can be used in following studies.