Elevated PD-L1 Expression and Microsatellite Instability in Elderly Patients With Gastric Cancer.

Immunotherapy in combination with chemotherapy is the current treatment of choice for frontline programmed cell death ligand 1 (PD-L1)-positive gastric cancer. However, the best treatment strategy remains an unmet medical need for elderly or fragile patients with gastric cancer. Previous studies have revealed that PD-L1 expression, Epstein-Barr virus association, and microsatellite instability-high (MSI-H) are the potential predictive biomarkers for immunotherapy use in gastric cancer. In this study, we showed that PD-L1 expression, tumor mutation burden, and the proportion of MSI-H were significantly elevated in elderly patients with gastric cancer who were older than 70 years compared with patients younger than 70 years from analysis of The Cancer Genome Atlas gastric adenocarcinoma cohort [≥70/<70: MSI-H: 26.8%/15.0%, P =0.003; tumor mutation burden: 6.7/5.1 Mut/Mb, P =0.0004; PD-L1 mRNA: 5.6/3.9 counts per million mapped reads, P =0.005]. In our real-world study, 416 gastric cancer patients were analyzed and showed similar results (≥70/<70: MSI-H: 12.5%/6.6%, P =0.041; combined positive score ≥1: 38.1%/21.5%, P <0.001). We also evaluated 16 elderly patients with gastric cancer treated with immunotherapy and revealed an objective response of 43.8%, a median overall survival of 14.8 months, and a median progression-free survival of 7.0 months. Our research showed that a durable clinical response could be expected when treating elderly patients with gastric cancer with immunotherapy, and this approach is worth further study.

Development of chitosan/ß-glycerophosphate/glycerol hydrogel as a thermosensitive coupling agent.

This work develops a dual-function thermosensitive hydrogel to prevent overheating, a side effect of focused ultrasound therapy. The proposed hydrogel has the components of chitosan, ß-glycerophosphate, and glycerol. Its thermosensitive sol-to-gel transition gives an instant signal of overheating without the need of any awkward sensing device. Impacts of varying component concentrations on the sol-to-gel temperature, rate, and degree of transparency are also investigated. Chemical structures and ultrasonic coefficients after heating are obtained with a Fourier transform infrared spectroscopy and ultrasonic measurement, respectively. Optimized formula of the proposed hydrogel is 0.5% chitosan, 5% ß-glycerophosphate, and 25% glycerol. This hydrogel has a high acoustic impedance (Z=1.8 Mrayl) close to that of human skin, high ultrasonic transmission (T=99%, which is normalized to water) from 25 to 55°C, and low attenuation coefficient (α=4.0Np/m). These properties assure the success of dual functions of the hydrogel developed in this work.

Advantages of magnetic resonance imaging (MRI) of the seminal vesicles and intra-abdominal vas deferens in patients with congenital absence of the vas deferens.

OBJECTIVE: To show the flexibility in magnetic resonance imaging (MRI) of seminal vesicle (SV) and intra-abdominal segment of vas deferens for the patients with congenital absence of the vas deferens (CAVD), including congenital bilateral absence of the vas deferens (CBAVD) and congenital unilateral absence of vas deferens (CUAVD). METHODS: Fourteen consecutive patients with CAVD had transrectal ultrasonography (TRUS) and further MRI evaluations. TRUS was performed using a 7.5-MHz transducer, and images of the SVs were obtained, calculated in the transaxial plane. MRI studies were performed with a 1.5-7 superconducting system, T1- and T2-weighted axial, coronal, and sagittal imaging of the pelvis was obtained. If the SVs were present, then their size was measured for the morphologic classification and diagnosis. All of the patients also received cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation testing. RESULTS: In a series of 12 men with CBAVD, only 4 were found to have bilateral SV agenesis using MRI. The remaining 8 men with unilateral hypoplasia still had SV remnants. MRI also detected the intra-abdominal segment of the vas deferens. Through our study of MRI, SV agenesis is not well associated with the presence of CFTR mutation in patients with CAVD. CONCLUSION: MRI provides a precise imaginal diagnosis of SV defect, which is superior to the TRUS examination for the patients with CAVD. Compared with the previous inaccurate examination method of TRUS, this study demonstrates that MRI can provide better images for the patients with CAVD for the clinical diagnosis of existing defects of internal seminal tract and internal organs.

18F-fluorodeoxyglucose positron emission tomography for the initial evaluation and monitoring of therapeutic response in bilateral adrenal histoplasmosis.

(18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) of a 74-year-old man showed high FDG uptake in bilateral adrenal histoplasmosis. Four months after the administration of an appropriate antifungal treatment, we observed a significant decrease in FDG uptake into the lesions, with a 40% reduction in activity (maximal standardized uptake value). This imaging result indicated partial resolution of the disease and was consistent with clinical outcome. Our study results suggest that FDG-PET is a useful modality for initial whole-body evaluation, selection of an appropriate antifungal treatment regimen, and monitoring of therapeutic response in bilateral adrenal histoplasmosis.

Curcumin inhibits the proliferation of human hepatocellular carcinoma J5 cells by inducing endoplasmic reticulum stress and mitochondrial dysfunction.

Curcumin (diferuloylmethane), which is obtained from turmeric, the rhizome of Curcuma longa (L.), inhibits many human cancer cells. However, the molecular mechanisms responsible for curcumin-induced endoplasmic reticulum stress in human hepatic cellular carcinoma J5 cells, are not yet clearly understood. J5 cells were treated with various concentrations of curcumin for different durations. The cell viability was detected by MTT assay. The protein expressions of caspase-12, ATF6, GADD153, Calnexin, Calreticulin, PDI and Ero1-Lα, which are associated with endoplasmic reticulum stress and the unfolding protein response pathway, were examined by Western blot analysis. The cell cycle was analyzed by flow cytometry. The protein expressions of TCTP, Mcl-1, Bcl-2 and Bax, which are related to mitochondrial dysfunction, were detected by Western blot analysis. We also detected the ATF6 protein location by immunocytochemistry. The results showed that curcumin inhibits the proliferation of J5 cells in a time- and dose-dependent manner. Curcumin induced the unfolding protein response by down-regulating the protein expressions of Calnexin, PDI and Ero1-Lα and up-regulating the Calreticulin expression. Curcumin induces the GADD153 expression by cleaving caspase-12 and ATF6, and then by translocating ATF6 to the nucleus. Curcumin also down-regulates the protein expressions of TCTP, Mcl-1 and Bcl-2, in order to induce mitochondrial dysfunction. Curcumin induced cell cycle arrest at the G2/M phase by decreasing the Cdc2 expression. In conclusion, the present study showed that curcumin inhibits the proliferation of J5 cells by inducing endoplasmic reticulum stress and mitochondrial dysfunction.

Sann-Joong-Kuey-Jian-Tang up-regulates the protein expression of Fas and TNF-α in colo 205 cells in vivo and in vitro.

Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine prescription, has been used to treat lymph node diseases and tumors. However, the molecular mechanisms of SJKJT in human colon cancer in vivo and in vitro have not been clearly elucidated. In the present study, we investigated the molecular mechanisms of SJKJT in human colon cancer colo 205 cells in vitro and in vivo. In the in vitro study, colo 205 cells were treated with various concentrations (0.5, 1 and 2 mg/ml) of SJKJT. The protein expression of TNF-α, Caspase-8 and Caspase-3 in colo 205 cells was measured by Western blotting. The results demonstrate that SJKJT up-regulated Fas, TNF-α, Caspase-8 and Caspase-3 protein expression. In the in vivo study, human colon cancer colo 205 cells (3x106/0.2 ml) were injected subcutaneously into the flank area of nude SCID mice (n=32) randomly divided into four groups. SJKJT was dissolved in saline and then administered orally to the mice at concentrations of 0.01, 0.1 and 0.3 g/kg/day for 30 days. The control group was treated with an equal volume of saline. SCID mice were sacrified by CO2 inhalation and the xenograft tumors were dissected. Subsequently, the protein expression of Fas, TNF-α, Caspase-8 and Caspase-3 in the tumors was measured by Western blotting. The results demonstrate that SJKJT up-regulated Fas, TNF-α, Caspase-8 and Caspase-3 protein expression, both in vitro and in vivo. These observations suggest that SJKJT has therapeutic potential in colon cancer.

Anti-tumor activity of Sann-Joong-Kuey-Jian-Tang alone and in combination with 5-fluorouracil in a human colon cancer colo 205 cell xenograft model.

Malignant tumors are the leading cause of death in Taiwan; among these, colon cancer ranks third as a cause of cancer-related death. Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicinal prescription, has been used to treat lymph node diseases and infectious lesions, and exhibits cytotoxic activity in many cancer cell lines. Our previous studies demonstrated that SJKJT inhibits the proliferation of human colon cancer colo 205 cells in vitro. The aim of this study was to evaluate the anti-tumor activity of SJKJT alone and in combination with 5-fluorouracil (5-FU) in vivo. SCID mice bearing human colon cancer colo 205 cell xenografts were administered SJKJT alone (30 mg/kg daily, p.o.), SJKJT (30 mg/kg daily, p.o.) in combination with 5-FU (30 mg/kg weekly, i.p.), or vehicle alone. At the end of the 4-week dosing schedule, the tumor and animal body weights were individually measured. The SCID mice were sacrificed with CO2 inhalation, the xenograft tumors were dissected, and the protein expression of microtubule-associated protein light chain 3 (MAP-LC3-II) in colo 205 xenograft tumors was measured by Western blotting. In the control, SJKJT-, and SJKJT plus 5-FU-treated mice, the tumor weights were 6.37±2.57, 0.43±0.35 and 1.63±0.46 g, and the mice body weights were 29±0.55, 29±2.71 and 27±0.77 g, respectively. Treatment with SJKJT resulted in a reduction in tumor weight compared with the control group, indicating that SJKJT inhibits tumor growth in a colo 205 xenograft model. SJKJT also increased LC3-II protein expression as compared to the controls. The present study shows that SJKJT alone or in combination with 5-FU has a positive effect on the treatment of SCID mice bearing human colon cancer colo 205 cell xenografts. This suggests that SJKJT has therapeutic potential in the treatment of human colon cancer.

Sann-Joong-Kuey-Jian-Tang increases the protein expression of microtubule-associated protein II light chain 3 in human colon cancer colo 205 cells.

Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine, has been prescribed as complementary medication for colon cancer in Taiwan. However, its molecular mechanisms are not yet understood. In the present study, we investigated the effects of SJKJT on human colon cancer colo 205 cells in vitro. The cytotoxicity of SJKJT in colo 205 cells was evaluated using the MTT assay, and the protein expression of microtubule-associated protein II light chain 3 (MAP-LC3-II) was measured using Western blot analysis. The results showed that SJKJT inhibited the survival rates of colo 205 cells in a time- and dose-dependent manner, with an IC50 concentration at 24 h of 590.34 µg/ml. In addition, SJKJT up-regulated the protein expression of MAP-LC3-II in colo 205 cells. These findings indicate that one of the molecular mechanisms by which SJKJT inhibits the proliferation of colo 205 cells in?vitro may be through the induction of the autophagic pathway. SJKJT may therefore have therapeutic potential for the treatment of human colon cancer.

Tanshinone IIA down-regulates the protein expression of ErbB-2 and up-regulates TNF-alpha in colon cancer cells in vitro and in vivo.

Tanshinone IIA (Tan-IIA) was isolated from Salviae Miltiorrhizae Radix. Our previous studies showed that Tan-IIA induced apoptosis in human colon cancer colo 205 cells, but the molecular mechanisms of the effect of Tan-IIA on human colon cancer were not clearly elucidated. The protein expression of ErbB-2 was up-regulated and activated in human and experimental colon cancers. In the present study, the effects of Tan-IIA on the protein expression of ErbB-2 in colo 205 cells were investigated. In vitro, colo 205 cells were treated with various concentrations of Tan-IIA (1, 2 and 5 mug/ ml) for 24 h, and the protein expression of TNF-alpha, ErbB-2 and caspase-3 was assayed by Western blotting. For the in vivo studies, male SCID mice were xenografted with colo 205 cells, and from day 10, Tan IIA (20 mg/kg/day, dissolved in corn oil) was administered by oral feeding for 30 days. As a control, mice with xenografted tumors were separately treated with corn oil (0.1 ml/10 g body weight). Expression of TNF-alpha, ErbB-2 and caspase-3 proteins was measured by Western blot analysis. Our results showed that Tan-IIA down-regulated the protein expression of ErbB-2 and up-regulated TNF-alpha and caspase-3 in colo 205 cells in vitro. In a colo 205 xenograft model, treatment with Tan-IIA caused up-regulation of TNF-alpha, caspase-3 and down-regulation of ErbB-2 protein expression as compared to the controls. Based on these observations, one possible molecular mechanisms by which Tan-IIA inhibits the proliferation of colo 205 cells is through the down-regulation of ErbB-2 protein expression and the up-regulation of the protein expression of TNF-alpha and caspase-3.

Tanshinone IIA inhibits human breast cancer cells through increased Bax to Bcl-xL ratios.

Tanshinone IIA (C19H18O3) was extracted from danshen (Salviae miltiorrhizae radix). It has cytotoxic properties and induces apoptosis in many human cancer cells. The molecular mechanisms are poorly understood, therefore, in the present study, we aimed to elucidate its anticancer activity on human breast cancer MDA-MB-231 cells. The cytotoxic effects of tanshinone IIA on MDA-MB-231 cells were measured by MTT assay. The percentages of cells in different cell cycle phases were determined by flow cytometry. The protein expression of Bax and Bcl-2 was examined using Western blotting. The results showed that tanshinone IIA inhibits the proliferation of MDA-MB-231 cells in a dose- and time-dependent manner. Tanshinone IIA induces apoptosis in a dose-dependent manner and the percentage of cells in sub-G1 phase. It increases the protein expression of Bax but decreases the Bcl-2 expression in MDA-MB-231 cells. Our findings suggest that tanshinone IIA can inhibit the proliferation of MDA-MB-231 cells by active apoptosis. One of the mechanisms may be through up-regulating the expression of Bax but down-regulating Bcl-2 expression and then inducing apoptosis. In conclusion, tanshinone IIA has therapeutic potential in breast cancer patients.

Diagnostic performance of a random versus lesion-directed biopsy of the prostate from transrectal ultrasound: results of a 5-year consecutive clinical study in 1 institution in South Taiwan.

OBJECTIVE: The purpose of this study was to compare cancer detection rates of sonographically suspicious lesion-directed biopsies and random biopsies through transverse examination of prostate halves in 518 patients over 5 years. METHODS: From 1998 to 2002, 518 patients were referred for prostate biopsies because of either elevated prostate-specific antigen (PSA) levels or abnormal digital rectal examination findings. On the basis of transverse examination of prostate halves by transrectal ultrasound, we performed lesion-directed biopsy of 3 to 5 cores if morphologically suspicious lesions existed on sonography or random biopsy of 3 cores if no obvious suspicious lesions existed. Biopsy specimens were put into 2 labeled containers. Pathologic results were correlated with random and lesion-directed guided locations. RESULTS: Nine-hundred fifty-nine results were obtained from 439 random and 520 lesion-directed biopsies in 518 patients. Cancer was detected in 207 patients (40.7%). Patients with cancer who had PSA levels of 10.0 ng/mL or greater had higher proportions of bilateral cancer lesions than those with PSA levels of less than 10 ng/mL (P = 0.03). One hundred ten (25.1%) of 439 normal-appearing halves taken by random biopsy were tumor-positive compared with 200 (38.5 %) of 520 biopsies from halves with sonographically suspicious lesions. Regardless of having random or lesion-directed biopsies, patients with PSA levels of 10 ng/mL or greater had higher positive malignancy rates than those with PSA levels of less than 10.0 ng/mL (P < .001). In about 40 patients, a diagnosis was made by random biopsy from halves that were morphologically normal on sonography, not by lesion-directed biopsy from tumor-suspicious contralateral halves. CONCLUSIONS: Cancer detection rates of lesion-directed biopsies are superior to those of random biopsies regardless of PSA level.