ALDH1A1 overexpression is associated with the progression and prognosis in gastric cancer.

BACKGROUND: Aldehyde dehydrogenase 1 family member A1 (ALDH1A1) is a cancer stem cell marker, and its expression correlates with prognosis in a number of malignancies. The aim of this study is to determine the relationship of ALDH1A1 expression with clinicopathological parameters and prognosis in gastric cancer. METHODS: ALDH1A1 and matrix metallopeptidase 9 (MMP-9) was evaluated by immunohistochemistry in 216 gastric carcinoma samples. The association between expression of ALDH1A1 and MMP-9, clinicopathological parameters, and prognosis of gastric cancer was examined. RESULTS: ALDH1A1 protein expression was significantly associated with depth invasion, lymph node metastasis and stage of disease (all P<0.05). Both univariate and multivariate analyses revealed that ALDH1A1 was an independent prognostic factor for both overall survival (OS) and recurrence-free survival (RFS) (both P<0.001). Furthermore, ALDH1A1 overexpression was associated with poor prognosis in patients subgroups stratified by tumor size, depth invasion and lymph node metastasis. Moreover, ALDH1A1 was significantly correlated with MMP-9 among 216 gastric cancer tissues (P<0.001). Patients who had ALDH1A1 overexpression, in which tumor cells displayed high invasiveness, had poor OS and shorter RFS. CONCLUSION: ALDH1A1 plays an important role in tumor aggressiveness and prognosis, and may act as a promising target for prognostic prediction.

Heat shock protein 60 overexpression is associated with the progression and prognosis in gastric cancer.

BACKGROUND: Heat shock protein 60 (HSP60) is a chaperonin with essential functions for cell physiology and survival, and its expression correlates with prognosis in a number of malignancies. The aim of this study is to determine the relationship of HSP60 status with clinicopathological parameters and prognosis in gastric cancer. METHODS: The levels of HSP60 and matrix metallopeptidase 9 (MMP-9) antigen was evaluated by immunohistochemistry in 223 gastric carcinoma samples. The association between HSP60 and MMP-9, clinicopathological parameters, and prognosis of gastric cancer was examined. RESULTS: The level of HSP60 protein was significantly associated with depth invasion, lymph node metastasis and stage of disease (all P<0.05). Both univariate and multivariate analyses revealed that HSP60 was an independent prognostic factor for both overall survival (OS) and recurrence-free survival (RFS) (both P<0.05). Furthermore, HSP60 overexpression was associated with a poor prognosis in patients with advanced gastric cancer in different risk groups. Moreover, HSP60 was significantly correlated with MMP-9 among 223 gastric cancer tissues (P<0.001). Patients who had HSP60 overexpression, in which tumor cells displayed high invasiveness, had poor OS and shorter RFS. CONCLUSION: HSP60 plays an important role on tumor aggressiveness and prognosis, and may act as a promising target for prognostic prediction.

Heat shock protein 22 overexpression is associated with the progression and prognosis in gastric cancer.

PURPOSE: The heat shock protein 22 (HSP22) is associated with tumor proliferation and protects tumor cell from apoptosis in many malignancies. However, the role of HSP22 in gastric cancer has not been thoroughly elucidated. The aim was to determine the relationship of HSP22 expression with clinicopathological parameters and prognosis in gastric cancer and estimate the alteration of HSP22 expression after neoadjuvant chemotherapy. METHODS: HSP22 and matrix metallopeptidase 9 (MMP-9) antigen expressions were evaluated by immunohistochemistry in 129 gastric carcinoma samples. Univariate and multivariate analyses were performed to determine the association between HSP22 expression and prognosis. The response of HSP22 was assessed in 47 patients who received neoadjuvant chemotherapy. RESULTS: HSP22 protein expression was significantly associated with tumor size, depth invasion, lymph node metastasis and stage of disease (all P < 0.05). In univariate and multivariate analyses, HSP22 was an independent prognostic factor for both overall survival (OS) and recurrence-free survival (RFS) (P = 0.003 and P = 0.004, respectively). Furthermore, HSP22 overexpression was associated with a poor prognosis in all patients and in patients subgroups stratified by tumor size, depth invasion and lymph node metastasis. In addition, HSP22 was significantly correlated with MMP-9 among 129 gastric cancer tissues (P < 0.001). Patients who had MMP-9 overexpression had poor OS and shorter RFS. Moreover, the alteration of HSP22 was not comparable in 47 patients who underwent neoadjuvant chemotherapy. CONCLUSIONS: HSP22 plays an important role on tumor aggressiveness and prognosis and may act as a promising target for prognostic prediction.

Clinical effect of erlotinib as first-line treatment for Asian elderly patients with advanced non-small-cell lung cancer.

PURPOSE: To evaluate the efficacy and toxicities of erlotinib as first-line treatment for Asian elderly patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Untreated patients with advanced NSCLC were included in this study; erlotinib was orally administered at a dose of 150 mg daily until disease progression or intolerable toxicity or for other reasons. RESULTS: A total of 35 patients were enrolled. Patient characteristics were as follows: mean age 75.6 years (ranged 70-81 years), 24 (68.6%) male, 16 (45.7%) former or current smokers, 13 (37.1%) adenocarcinoma, 18 (51.4%) squamous cell carcinoma and 4 (11.4%) bronchioloalveolar carcinoma. Out of 35 patients, 1 CR, 16 PR and 10 SD, resulting in an overall response rate (CR + PR) of 48.6% and disease control rate (DCR = CR + PR + SD) of 77.1%. The median TTP was 6.4 months, and the median OS was 12.7 months. The CBR was 80%, and the 1-year survival rate was 48.6%. The most common adverse event (AE) was mild skin rash and diarrhea (CTC AE 1/2). Among them, the female never smokers with a non-squamous cell carcinoma histology was superior to the male smokers with a squamous cell carcinoma in disease control rate, with significant differences (P < 0.05). CONCLUSION: The results suggest that erlotinib monotherapy is an effective and well-tolerated treatment option for Asian elderly patients with advanced NSCLC.

Extracellular accumulation of recombinant protein by Escherichia coli in a defined medium.

Extracellular accumulation of recombinant proteins in the culture medium of Escherichia coli is desirable but difficult to obtain. The inner or cytoplasmic membrane and the outer membrane of E. coli are two barriers for releasing recombinant proteins expressed in the cytoplasm into the culture medium. Even if recombinant proteins have been exported into the periplasm, a space between the outer membrane and the inner membrane, the outer membrane remains the last barrier for their extracellular release. However, when E. coli was cultured in a particular defined medium, recombinant proteins exported into the periplasm could diffuse into the culture medium automatically. If a nonionic detergent, Triton X-100, was added in the medium, recombinant proteins expressed in the cytoplasm could also be released into the culture medium. It was then that extracellular accumulation of recombinant proteins could be obtained by exporting them into the periplasm or releasing them from the cytoplasm with Triton X-100 addition. The tactics described herein provided simple and valuable methods for achieving extracellular production of recombinant proteins in E. coli.

Extracellular production of human parathyroid hormone as a thioredoxin fusion form in Escherichia coli by chemical permeabilization combined with heat treatment.

A pET system encoding the fusion protein gene of thioredoxin (Trx) and human parathyroid hormone (hPTH) was introduced into Escherichia coli BL21 (DE3). Recombinant Trx-hPTH fusion protein was expressed in soluble form in the cytoplasm of the E. coli transformant. To recover Trx-hPTH from the E. coli culture efficiently, a novel tactic was developed by adding Triton X-100 into the fermentation culture at the exponential growth phase of E. coli and by heat treatment of the culture at the end of the fermentation. A concentration of 1% (v/v) Triton X-100 was added into the culture at the same time as IPTG addition after optimization. Under these conditions, addition of Triton X-100 had little effect on the cell growth, but more than 75% of the total recombinant Trx-hPTH was released into the fermentation broth. Also, a much higher volumetric yield of recombinant Trx-hPTH could be obtained with protein release compared to yield without protein release. Simultaneously, owing to the highly thermal stability of Trx-hPTH fusion protein, heat treatment of the fermentation broth at 80 degrees C for 15 min at the end of fermentation was employed for primary purification. Results demonstrated that heat treatment not only boosted further release of the recombinant Trx-hPTH fusion protein into the fermentation broth but also precipitated/denatured most of the nontarget proteins released in the broth. The tactics described herein integrated the fermentation process with subsequent recovery steps and thus provided a valuable and economical method for the production of Trx-hPTH and maybe some other Trx fusions in E. coli.

[Effects of aging and hypoxia on the proliferation of cultured rat pulmonary arterial smooth muscle cells].

OBJECTIVE: To study the effects of aging and hypoxia on the proliferative behavior of cultured pulmonary arterial smooth muscle cells (PASMCs). METHODS: PASMCs isolated from aged (18-24 months) and young (3-4 months) rats were divided, according to the different treatments the cells were subjected to, into young and aged normoxic groups (groups A and B) and young and aged hypoxic groups (groups C and D) respectively. MTT cell proliferation assay, 3H-TdR incorporation assay, flow cytometriy and immunocytochemical analysis were respectively employed to observe the proliferative behavior. RESULTS: Compared with the cells from young rats, PASMCs from aged rats had a higher proliferation rate, more 3H-TdR incorporation, increased mitotic cell ratio, reduced amount of the total protein, and elevated content of proliferating cell nuclear antigen (PCNA). In comparison with normoxic condition, hypoxia elicited higher proliferation rate of the cells with inhibition of 3H-TdR incorporation that was subsequently increased. Higher percentage of mitotic cells, less total protein amount and increased PCNA were also observed in response to hypoxia. CONCLUSIONS: Aging and hypoxia may directly induce PASMC proliferation, and in aging PASMCs, the proliferation is the most obvious in response to hypoxic stimulation.

Endothelial NO synthase gene expression in experimental vasospasm in rabbits.

OBJECTIVE: To investigate the relationship between endothelial NO synthase (eNOS) gene expression and experimental vasospasm. METHODS: Sixteen rabbits were randomly divided into 2 groups, which were subjected to balloon endothelial denudation with normal diet (n=8) or with hypercholesterol diet group (n=8). Angiography was performed to detect the vasospasm induced by ergonovine before and after denudation and 8 weeks after hypercholesterol feeding. In situ hybridization and Northern blotting were performed to localize and quantitate respectively the expression of eNOS mRNA. RESULTS: Visible vasospasm was induced at the denuded sites in rabbits with hypercholesterol diet, in which the expression of eNOS mRNA was detected in the endothelium by in situ hybridization at a lower level than that of rabbits with normal diet, as demonstrated by Northern blotting. CONCLUSION: The decrement of eNOS mRNA expression resulted from balloon endothelial denudation and hypercholesterolemia may play an important role in the pathogenesis of experimental vasospasm.

Changes in cardiac function reserve after percutaneous balloon mitral valvuloplasty: a longterm follow-up study.

OBJECTIVE: To evaluate the long-term effect of percutaneous balloon mitral valvuloplasty (PBMV) by observing the changes in mitral valve reserve after PBMV and during the long-term follow-up and by exploring the relations of these changes with stress echocardiographic scores. METHODS: Stress echocardiogaphy was performed in patients receiving PBMV for mitral stenosis both after surgery and during the long-term follow-up study, and Wilkins scores of the patients were obtained pre-operatively. Intravenous isoproterenol was administered before test to increase the heart rate to simulate mild, moderate and heavy exercises, and the indices for cardiac function were recorded. RESULTS: After PBMV, mitral valve reserve and cardiac function reserve were partly restored, and stress echocardiographic score was closely related to long-term cardiac function ( = 0.82). CONCLUSIONS: PBMV can increase mitral valve area and partly resotreit funtional reserve, and stress echocardiographic score is more predictive of the long-term cardiac function than Wilkins score.