FAST1 Predicts Poor Survival of Renal Carcinoma and Promotes Its Progression Through the TGF-ß/Smad Pathway.

PURPOSE: Renal carcinoma (RC) originates in the renal tubular epithelial system, among which renal cell carcinoma (RCC) is the most frequent one. The forkhead activin signal transducer 1 (FAST1) has been shown to interfere with tumor progression as an oncogene, while its role in RC is limited. Therefore, this paper explored the prognostic significance, specific effects, and related mechanisms of FAST1 on RC. PATIENTS AND METHODS: Cell colony formation assay, cell counting kit-8 (CCK8) assay, flow cytometry and Transwell assay were used to test cell proliferation, viability, apoptosis, migration and invasion, respectively. Western blot (WB) was employed to determine the protein level of FAST1. RESULTS: Our study confirmed that FAST1 was up-regulated in RC tissues and cell lines, and its overexpression often represented a poor prognosis of RC patients. Meanwhile, the in vitro experiments showed that overexpressing FAST1 facilitated RC cell viability, proliferation, migration, invasion and epithelial-mesenchymal transition (EMT), and repressed cell apoptosis. In addition, the in vivo experiments illustrated that the up-regulation of FAST1 strengthened tumor growth. On the contrary, knocking down FAST1 had the opposite effects. Mechanistically, The TGF-ß/Smad pathway contributed to RC evolvement and was activated by FAST1 both in vitro and in vivo. CONCLUSION: This article suggests that FAST1 exerts a carcinogenic role in RC by regulating the TGF-ß/Smad signaling.

Simultaneous Gd2O3 clusters decoration and O-doping of g-C3N4 by solvothermal-polycondensation method for reinforced photocatalytic activity towards sulfamerazine.

To improve the visible-light photocatalytic activity of graphitic carbon nitrate (g-C3N4) for practical application, a Gd2O3-cluster decorated O-doping g-C3N4 was fabricated via an ethanol assisted solvothermal-polycondensation method. The as-prepared photocatalysts, including bulk g-C3N4 (CN), O-doping g-C3N4 (HECN) and Gd2O3-cluster decorated O-doping g-C3N4 (HECN-xGd), were characterized and the paralleled experiments were conducted to evaluate the photocatalytic activity, mineralization capacity and mineralization mechanism, where sulfamerazine (SMR) was employed as the target pollutant. Furthermore, the quenching tests with scavengers were executed to analyze the contributions of the dominant active species, where the O2- was identified as a major role, and h+ as the minor role in the photodegradation of SMR. Results from the paralleled experiments suggested that the HECN-xGd possesses superior photocatalytic activity to HECN and CN, besides the feasible reusability through five cycles and impressive total organic carbon (TOC) removal about 60%. And the improved photocatalytic activity of HECN-xGd is ascribed mainly to the oxygen doping and Gd2O3 decoration. Herein, oxygen doping optimizes the structure of g-C3N4 and expands the light absorption range of HECN, and Gd2O3 facilitates the reduction of O2 into O2-, and acts as the separator and transporter for the photo-induced charges.

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.

[Effects of celecoxib on the proliferation and apoptosis of human nasopharyngeal carcinoma cell line CNE-2].

OBJECTIVE: To detect the effect of Celecoxib on the proliferation and apoptosis of human nasopharyngeal carcinoma cell line CNE-2. METHOD: The growth inhibition rate of CNE-2 by Celecoxib was evaluated with MTT method. Apoptosis related morphology changes were observed with transmission electron microscopy (TEM). The cell cycle and apoptosis were measured with flow cytometric method (FCM). Apoptotic index (AI) was counted by the TDT-mediated dUTP-biotin nick end-labeling (TUNEL) assay. RESULT: The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner. Apoptosis with nuclear chromatin condensation, cell shrinkage, periplasm loss and the formation of apoptotic bodies was observed with TEM. Apoptotic rates of CNE-2 cells treated with 80 and 100 micromol/L celecoxib were (10.47+/-0.18)% and (20.17+/-0.55)% respectively, significantly higher than those of the control group (1.57+/-0.27)% with FCM. The percentage of G0/G1 phase cells increased, whereas the S and G2/M phases cells decreased in a dose-dependent manner after the treatment. TUNEL assay showed that the apoptosis ratio (AI) of CNE-2 treated with Celecoxib was higher than control group (P<0.01). CONCLUSION: Celecoxib can inhibit the growth of human nasopharyngeal carcinoma cell line CNE-2 and induce the cell apoptosis, which may be related to blocking the cell cycle progress of CNE-2 cells.

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.

Pump models assessed by transesophageal echocardiography during cardiopulmonary resuscitation.

OBJECTIVE: Transesophageal echocardiography was performed during closed-chest cardiopulmonary resuscitation (CPR) in in-hospital cardiac arrest to further explore the hemodynamic mechanism of CPR. METHODS: CPR attempts were performed according to advanced cardiovascular life support guidelines in 6 cases of in-hospital cardiac arrest. Multi-plane transesophageal echocardiography was carried out within 15 min of initiation of CPR. Throughout CPR, the motion of the mitral, tricuspid and aortic valves, the changes in the left ventricular cavity size and the thoracic aortic diameter were observed. Trans-mitral and trans-aortic Doppler files of blood flow were also documented. RESULTS: A closure of the mitral and tricuspid valves with simultaneous opening of the aortic valve occurred exclusively during chest compression, resulting in forward blood flow in the pulmonary and systemic circulation. Peak forward aortic flow at a velocity of 58.8 +/- 11.6 cm/s was recorded during the compression phase. Whereas, a closure of the aortic valve and rapid opening of the atrioventricular valves associated with ventricular filling during relaxation of chest compression was noted in all 6 patients. Peak forward mitral flow at a velocity of 60.6 +/- 20.0 cm/s was recorded during the release phase. Mitral regurgitation during the chest compression period was detected in 5 patients, reflecting a positive ventricular-to-atrial pressure gradient. A reduction in the left ventricular chamber and an increase in the thoracic aortic diameter during the compression phase was found in all patients, indicating that direct cardiac compression contributed to forward blood flow. CONCLUSION: These observations favor the cardiac pump theory as the predominant hemodynamic mechanism of forward blood flow during CPR in human beings.

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.