Despite numerous epidemiological data linking type 2 diabetes mellitus (T2DM) and breast cancer (BCa), there is limited experimental evidence of this association. The clinically relevant question is at what stage diabetes may exert its tumor-promoting activity. Moreover, identification of major pathophysiological pathways underlying this activity should provide valuable information for treatment. In the present study, the BCa cells isolated from long-term T2DM-treated tumors from diabetic nude mice were found to have increased cell proliferation, invasiveness and docetaxel (DTX) resistance. Importantly, this stimulatory effect was only observable in estrogen receptor (ER)-positive BCa cells. Mechanistically, T2DM-elicited hyperinsulinemia induced HIF-1α expression by reducing its ubiquitination, which was accompanied with upregulated oxidative stress. Furthermore, in vivo inhibition of HIF-1α expression effectively reversed the above-mentioned tumor-promoting activity and partially attenuated T2DM-elicited oxidative stress. Altogether, the results provide novel and compelling experimental evidence that (i) prolonged exposure to T2DM promotes BCa progression; (ii) the hyperinsulinemia/HIF-1α/oxidative stress cascade is the major mediator of this effect.

OBJECTIVES:miRNAs play crucial roles in human tumourigenesis. This study was performed to measure expression and function of miR-762 in breast cancer.MATERIALS AND METHODS:Expression of miR-762 in breast tissues and cell lines (SK-BR-3, DA-MB-435s, MCF-7 and MDA-MB-231, HBL-100) was measured by using real-time RT-PCR. We restored expression of miR-762 in MCF-7 cells to measure its functional roles. Luciferase assays were performed to reveal the target gene of miR-762.RESULTS:Expression of miR-762 was high in both breast cancer cell lines and specimens, and its overexpression increased breast cancer cell proliferation and invasion. Interferon regulatory factor 7 (IRF7) is a direct target of miR-762 and overexpression of miR-762 reduced expression of IRF7. Moreover, IRF7 was repressed, its levels inversely correlated to miR-762 expression. IRF7 rescued miR-762-induced cell invasion and proliferation.CONCLUSIONS:These results demonstrate that miR-762 tumour effect was achieved by targeting IRF7 in human breast cancer specimens.

Membrane type 1 matrix metalloproteinase (MT1-MMP) has been demonstrated to play an important role in tumor progression. The aim of the present study was to analyze the expression of MT1-MMP in breast cancer and its correlation with clinicopathologic characteristics, including the survival of breast cancer patients. In our results, MT-MMP1 was up-expressed in breast cancer tissues compared with ductal hyperplasia tissues in microarray data (GSE2429). MT1-MMP mRNA and protein expression was markedly higher in breast cancer tissues than in normal breast tissues (P=0.005 and P=0.037, respectively). Using immunohistochemistry, high levels of MT1-MMP protein were positively correlated with the status of clinical stage (I-II vs. III-IV; P=0.043), lymph node metastasis (absence vs. presence; P=0.024), and distant metastasis (No vs. Yes; P=0.017) of breast cancer patients. Patients with higher MT1-MMP expression had a significantly shorter overall survival time than did patients with low MT1-MMP expression. Multivariate analysis indicated that the level of MT1-MMP expression was an independent prognostic indicator (P<0.001) for the survival of patients with breast cancer. In conclusions, MT1-MMP plays an important role on breast cancer aggressiveness and prognosis and may act as a promising target for prognostic prediction.

Patchouli alcohol (PA), a tricyclic sesquiterpene isolated from Pogostemonis Herba, has been known to exhibit antioxidant, anti-inflammatory, and other important therapeutic activities. The aim of this study was to investigate the effects of PA on LPS-induced mastitis in vivo and the possible mechanism. The mouse model of mastitis was induced by injection of LPS through the duct of mammary gland. Mice were pretreated with dexamethasone or PA 1 h before and 12 h after induction of LPS. The myeloperoxidase activity and inflammatory cytokines production in mammary tissues were determined. The effects of PA on NF-κB signal pathways were analyzed by Western blotting. The results showed that PA inhibited the LPS-induced TNF-α, IL-6, and IL-1β production in a dose manner. It was also observed that PA attenuated mammary histopathologic changes. Furthermore, Western blot analysis showed that PA could inhibit the phosphorylation of NF-κB and IκB induced by LPS. These results indicate that PA inhibits NF-κB signaling pathways to attenuate inflammatory injury induced by LPS. PA may be a potent therapeutic reagent for the prevention of mastitis.

Gastric cancer (GC) is one of the most threatening diseases. The symptoms of GC are complex and hard to detect, which also contribute to the poor prognosis of GC. Besides, the current diagnosis forGCis expensive and invasive. Thus, a fast, noninvasive biomarker is urgently needed for GC screening. MicroRNAs (miRNAs) are small noncoding RNAs, which are involved in a great variety of pathological processes, particularly carcinogenesis. MiRNAs are stable in gastric juice, plasma as well as serum, which facilitate it to be a promising biomarker for cancer. In this study, we selected three novel miRNAs, i.e., miR-233, miR-16, and miR-100, to investigate their potential diagnostic value in GC screening. A total of 50 GCpatients and 47 healthy controlswere involved in this study. Blood serum samples were collected; RNAswere extracted and normalized with U6 snRNA as the internal control; qRT-PCR was performed for relative expression of target miRNAs. Levels of miRNAs expression were compared by Student's t test for the comparison between two groups, and one-way ANOVAwas used for multiple comparisons. The expression of miR-223, miR-16, and miR-100 was all significantly higher in GC patients than controls (all P < 0.001). All the tested miRNAs were manifested to be valuable biomarkers for GC. Relative expression of these miRNAs was significantly correlated with clinical characteristics of GC patients, such as TNM stage (P = 0.036 for miR-223; P < 0.001 for miR-100), metastatic status (P = 0.045 for miR-223; P = 0.031 for miR-16; P = 0.006 for miR-100), tumor size (P = 0.042 for miR-223; P = 0.031 for miR-16; P < 0.001 for miR-100), and differentiation grade (P = 0.036 for miR-223; P = 0.030 for miR16; P = 0.034 for miR-100). However, in T classification, which considered both tumor size and direct extent of primary tumor, the difference in target miRNAs expression was not significant. In summary, we confirmed the diagnostic value of serum miR-223, miR-16, and miR-100 in GC. Significantly elevated expression of the three miRNAs was also observed in advanced GC patients, which suggested their availability in cancer staging.

Combating with multidrug resistance (MDR) is a major part of hepatocellular carcinoma (HCC) chemotherapy. Steroidal saponin from Trillium tschonoskii (TTS) could be a potential weapon. We found TTS could reverse the MDR in HCC cells and significantly enhance chemosensitization. TTS inhibited HepG2 and R-HepG2 cells survival in a dose-dependent manner by 75% and 76%, respectively (p<0.01), as well as colony formation 77% and 81% (p<0.01). Moreover, TTS induced sensitization of R-HepG2 to anti-cancer drugs, indicated by significantly reduced IC50. On the other hand, TTS suppressed expression of P-glucoprotein in MDR HCC cells, and thereby increased accumulation of doxorubicin from 126 ng/10(5)cells to 752 ng/10(5)cells (p<0.01). TTS also repressed expression of many other MDR genes, such as MRP1, MRP2, MRP3, MRP5, MVP and GST-π. In vivo, TTS dose-dependently reduced R-HepG2 cells xenografts tumour formation by inhibiting tumour cells proliferation in mice. Consistence with in vitro finding, TTS induced R-HepG2 sensitization to doxorubicin and therefore reduced tumour formation in vivo.

Objective: To observe the effects of chemosensitizer on expression of P-glycoprotein (P-gp) in multidrug-resistant (MDR) carcinoma cells and the changes of corresponding Tc-99(m)-MIBI (referring to methoxy isobutyl isonitrile) uptake kinetics. Methods: Firstly, the immunocytochemistry was used to test the experssion level of P-gp in carcinoma cells; then the 7 counter was used to test the uptake rate of radioactive precipitation of Tc-99(m)-MIBI; and finally to do statistical analysis of the test data. Results: The P-gp expression in MCF-7/S cells was negative, while it was strongly positive in MCF-7/ADM cells, and the P-gp expression in MCF-7/ADM cells reduced significantly after the tetrahydropalmatine (Tet) effect. The uptake rate of MCF-7/S cells reached the maximal value (1.39%) after being cultured with Tc-99(m)-MIBI for 90 minutes, and then it kept in equilibrium. The Tc-99(m)-MIBI uptake rate of MCF-7/ADM cells had been maintained at low level, being 0.371% after 90 minutes. The Tc-99(m)-MIBI uptake rate of MCF-7/S cells was higher than that of MCF-7/ADM cells. After the tetrahydropalmatine and verapamil effects, the Tc-99(m)-MIBI uptake rate of MCF-7/S cells had no significant change, while the Tc-99(m)-MIBI uptake rate of MCF-7/ADM cells increased by 1.9 times. Conclusion: Tet can reverse the MDR in MCF-7 cells, and the mechanism can achieve the effect of reversing the drug resistance through adjusting down the expression of P-gp in carcinoma cells.

BACKGROUND AND OBJECTIVES: Radiotherapy is frequently applied in the treatment of malignant gliomas, but it is unclear if radiotherapy exerts its effects via induction of apoptosis. The present study was designed to determine whether a single-fraction γ-60Co radiation can induce apoptosis. DESIGN AND SETTING: In vitro cytological controlled study performed at a military medical university from October 2006 to June 2008. METHODS: C6 cells were treated with a single fraction of γ-60Co radiation at various doses (0, 4, 16, and 64 Gy). The 3-(4,5)-dimethylthiazol-2)-2,5-diphenyl tetrazolium bromide (MTT) assay, apoptosis assays using Annexin V-fluorescein isothiocyanate/propidium iodide or Hoechst 33258 staining, and the cell cycle assay were performed, and the expression of p53 and p21 proteins was evaluated. RESULTS: The C6 cell numbers in the 16 Gy and 64 Gy groups were much lower than in the control group at 48, 96, and 144 hours after irradiation. The irradiated cells underwent apoptosis in a dose-dependent manner. Irradiation also impacted cell cycle progression, arresting cells in the G1 phase. The p53 protein expression was shown in both the nucleus and the cytoplasm of irradiated cells, whereas p53 was only expressed in the nucleus of control (untreated) cells. The p21 protein was expressed in irradiated cells but not in control cells. CONCLUSIONS: Single-fraction γ-60Co radiation inhibited C6 cell growth by inducing apoptosis and G1 arrest, which correlated with the up-regulation of the p53-p21 pathway. The extent of apoptosis and G1 arrest was positively correlated with the dose of radiation. Better understanding of apoptosis induced by radiation therapy will help design optimal dosing schedules for radiation therapy, especially in combination with chemotherapy.

Hepatoma-derived growth factor (HDGF) plays an important role in tumor progression. Highly expressed HDGF has been found to indicate poor prognosis in many cancers. However, no information is available regarding the prognostic value of nuclear or cytoplasmic HDGF staining level in breast cancer. In the present study, the nuclear or cytoplasmic HDGF staining level was investigated in 86 patients with primary breast cancer by immunohistochemistry; the relationship between nuclear or cytoplasmic HDGF staining level and clinicopathological parameters was examined by Two-tailed Mann–Whitney U-test or Krustal–Wallis. The prognostic value of nuclear or cytoplasmic HDGF staining level in disease-free survival and overall survival was analyzed by Kaplan–Meier methods and log-rank test. We found that the percentage of cases with strong nuclear HDGF staining level was significantly higher in the cases with high tumor grade, high stage, high proliferation index (Ki-67 index > 20%), as well as in those with lymph node invasion and recurrence (p < 0.05) compared to those without. No significant correlation was found between cytoplasmic HDGF expression and any clinicopathological variables. In addition, disease-free survival and overall survival were significantly lower in patients with high nuclear HDGF expression (level 2) than in those with low nuclear HDGF expression (level 0 and level 1). Further Cox multivariate analysis showed that high nuclear HDGF expression is an independent factor for indicating poor prognosis in breast cancer patients. No significant difference in disease-free survival rate and overall survival was found between different cytoplasmic HDGF staining levels. All these findings suggest that increased nuclear HDGF expression is involved in tumor progression and might be used as a new prognosticator for breast cancer.

Nodular histiocytic aggregate (NHA) of the omentum is a rare benign proliferative process composed predominantly of histiocytes with scattered mesothelial cells. NHA is a differential diagnosis for neoplasms or metastatic tumors in cancer patients. To further clarify this clinical pitfall issue, the authors investigated surgical samples of the greater omentum from 96 patients with gastrointestinal malignancies and 53 patients with gynecologic neoplasms. Visible NHA of greater omentum was identified in 3 patients with ovarian neoplasms (borderline mucinous cystadenoma, low-grade papillary serous cystadenocarcinoma, and juvenile granulosa-cell tumor) but in none of the patients with gastrointestinal malignancies. Similar lesion was also identified on the cell blocks from peritoneal washings in 1 of the 3 patients. Grossly, the lesions formed small yellow-red nodules on the greater omentum, and the NHA lesion was also found diffusely on the surface of the appendix and fallopian tubes in 2 of the 3 patients. Histological study showed that typical NHA changes over an inflammatory background, which may indicate that NHA is a consequence of a chronic inflammatory process of omentum. The predominant infiltration of T lymphocytes in the NHA lesions indicates that the aggregation of histiocytes may be related to the activation of T-cell immunity. This report has first demonstrated visible NHA in the greater omentum of patients with ovarian malignancies, and awareness of this entity should be brought to clinicians to avoid misdiagnosis.

Objective: To observe the effects of traditional Chinese drug Yiqiyangjing on the postoperative immune function in breast cancer patients. Method: 136 breast cancer patients who underwent modified radical mastectomy were randomly divided into treatment group and control group. The treatment group received a decoction of traditional Chinese medicine Yiqiyangjing since the 7th preoperative day and the 3rd postoperative day, while the control group received no additional traditional Chinese drug to the basic therapy same as the treatment group. Each immune index was determined on the 7th preoperative day as well as the 1st, 7th and 14th postoperative days respectively. Results: On the 14th postoperative day, immunoglobulin IgA, IgG and IgM of the treatment group were all significantly higher than those of the control group; CD4, CD4/CD8 and IL-2 were also significantly rebounded after 14-day treatment of traditional Chinese drug (P<0.05); and TNF-alpha was significantly increased after the surgery, and recovered to the preoperative level in the treatment group while remained at a high level in the control group after the 14th postoperative day. Conclusion: The traditional Chinese drug is beneficial to the postoperative recovery of humoral immune function and cellular immune function in breast cancer patients.

We investigated the relationship of End-to-end distance between VH and VL with different peptide linkers and the activity of single-chain antibodies by computer-aided simulation. First, we developed (G4S)n (where n = 1-9) as the linker to connect VH and VL, and estimated the 3D structure of single-chain Fv antibody (scFv) by homologous modeling. After molecular models were evaluated and optimized, the coordinate system of every protein was built and unified into one coordinate system, and End-to-end distances calculated using 3D space coordinates. After expression and purification of scFv-n with (G4S)n as n = 1, 3, 5, 7 or 9, the immunoreactivity of purified ND-1 scFv-n was determined by ELISA. A multi-factorial relationship model was employed to analyze the structural factors affecting scFv: rn=ABn-ABO2+CDn-CDO2+BCn-BCst2. The relationship between immunoreactivity and r-values revealed that fusion protein structure approached the desired state when the r-value = 3. The immunoreactivity declined as the r-value increased, but when the r-value exceeded a certain threshold, it stabilized. We used a linear relationship to analyze structural factors affecting scFv immunoreactivity.

Objective: To observe the effects of chemosensitizer on expression of P-glycoprotein (P-gp) in multidrug-resistant (MDR) carcinoma cells and the changes of corresponding 99Tcm-MIBI (referring to methoxy isobutyl isonitrile) uptake kinetics. Methods: Firstly, the immunocytochemistry was used to test the experssion level of P-gp in carcinoma cells; then the γ counter was used to test the uptake rate of radioactive precipitation of 99Tcm-MIBI; and finally to do statistical analysis of the test data. Results: The P-gp expression in MCF-7/S cells was negative, while it was strongly positive in MCF-7/ADM cells, and the P-gp expression in MCF-7/ADM cells reduced significantly after the tetrahydropalmatine (Tet) effect. The uptake rate of MCF-7/S cells reached the maximal value (1.39%) after being cultured with 99Tcm-MIBI for 90 minutes, and then it kept in equilibrium. The 99Tcm-MIBI uptake rate of MCF-7/ADM cells had been maintained at low level, being 0.371% after 90 minutes. The 99Tcm-MIBI uptake rate of MCF-7/S cells was higher than that of MCF-7/ADM cells. After the tetrahydropalmatine and verapamil effects, the 99Tcm-MIBI uptake rate of MCF-7/S cells had no significant change, while the 99Tcm-MIBI uptake rate of MCF-7/ADM cells increased by 1.9 times. Conclusion: Tet can reverse the MDR in MCF-7 cells, and the mechanism can achieve the effect of reversing the drug resistance through adjusting down the expression of P-gp in carcinoma cells.

Retinoblastoma (Rb) is the most common eye cancer in children and it can be inherited. Rb is quite rare and originators from the neural retina with a significant genetic component in etiology, which occurs in approximately 1 in every 20 0000 births. In children with the heritable genetic form of Rb, there is a mutation on chromosome 13, called the retinoblastoma 1 (Rb 1) gene. Early diagnosis and intervention is critical to the successful treatment of the Rb. The Rb1 gene is the first cloned tumor suppressor gene. As a negative regulator of the cell cycle, Rb1 gene could maintain a balance between cell growth and development through binding to transcription factors and regulating the expression of genes involved in cell proliferation and differentiation. Thus, it is involved in cell cycle, cell senescence, growth arrest, apoptosis and differentiation. We summarized the recent advances on the epidemiology and Rb1 gene of Rb in this review.

Summary 1. The aim of the present study was to evaluate the clinical value of colour Doppler application in encircling constriction of the superficial femoral vein in deep vein insufficiency. 2. A total of 87 patients with primary deep venous insufficiency (PDVI) using ascending venography were randomly divided into group A (44 patients) and group B (43 patients). All patients underwent encircling constriction of the superficial femoral vein, high ligation and ablation of the great saphenous vein and perforator vein. The duration of venous reflux at operation was monitored with colour Doppler in group A (but not group B) to evaluate the immediate effects. Clinical grading and scoring of the clinical, etiological, anatomical, pathophysiological (CEAP) classification system were used to evaluate the follow‐up curative effect. 3. In four cases from group A, completely destroyed valves were identified at the time of operation and autografting of the vein segment with a valve was carried out. The intraoperative examination of colour Doppler in group A showed a much shorter duration of vein reflux after the encircling constriction procedure than the presurgery condition. According to the results of CEAP grading, the success rate of group A (95.0%, 38/40) was significantly higher than that of group B (76.7%, 33/43). Postoperative clinical scores were markedly lower than preoperative scores in both groups A and B. 4. In conclusion, our data suggest that application of colour Doppler in encircling constriction of superficial femoral vein might enhance surgical pertinence and improve surgical effect for PDVI.

BACKGROUND:Intrathoracic anastomotic leakage resulted from radical total gastrectomy with an end-to-side esophagojejunostomy are exclusively abdominal.CASE PRESENTATION:We report the case of a 64-year-old male who underwent radical total gastrectomy and intraabdominal end-to-side esophagojejunostomy for gastric cardiac carcinoma. Anastomotic leakage to the thoracic cavity occurred which was confirmed by contrast radiography 18 days after the operation. The symptoms included coughing and fever, with elevated white blood cells over 10×10(9)/L. Coughing and fever disappeared after successful sealing of the fistulous orifice with an endoscopically placed covered metallic stent with the applications of antibiotics and drainage of the pleural effusion. The patient was recovered and discharged from the hospital approximately two months after the occurrence of the leakage without any symptoms except intermittent esophageal reflux which could be resolved by treatment with cisapride, or by intaking less liquid food. The patient then received 4 cycles of adjuvant chemotherapy with regimen of FOLFOX4 (fluorouracil, leucovorin and oxaliplatin). Unfortunately, he died of a disease- or treatment- unrelated accidence 5 months after the discharge.CONCLUSION:The thorough drainage combined with antibiotic treatment is able to eliminate empyema without the need for a specific thoracoscopy or thoracic surgery for patients with intrathoracic leakage.

scFv (single-chain Fv fragment) is the gene engineered antibody employed widely in at present. Recently there have been some progress on colorectal cancer diagnosis and treatmentoriented with scFv as carrier, but it is differential on affinity and anti-tumor activity of scFv with that of derived factor. Proper linker could make scFv have more suitable biological activity on clinical application. So it becomes a key step that how to choose and design a proper linker. In this work, the (Gly(4)Ser) n is used as the linker to connect VH and VL separately (n=0,1,2,3,4,5,6,7,8,9).and the 3D structure of corresponding scFv is reconstructed by homology modeling. The End-End distance of scFy protein with different linker (G(4)S) n, which 3D structure coordinate system are normalized, are compared to evaluate the effect of linker length to VH and VL. In molecular biology experiment, ND-1scFv-n are expressed and analyzed with n=1,3,5,7,9, their biologic activity active are consistent with what estimated by End-End distance. So the index to estimate biologic activity of scFv with its End-End distance is presented and could be helpful to further understand the relationship between the spatial structure of single-chain antibody and the physical and chemical properties.

Inflammatory bowel disease (IBD) includes two similar yet distinct conditions called ulcerative colitis (UC) and Crohn's disease (CD). These diseases affect the digestive system and cause the inflammation of intestinal tissue, form sores and bleed easily. Most children with IBD are diagnosed in late childhood and adolescence. However, both UC and CD have been reported as early as in infancy. Most information pertaining to the epidemiology of IBD is based upon adult studies. Symptoms include abdominal pain, cramping, fatigue and diarrhea. Genetic factors play a significant role in determining IBD susceptibility. Epidemiological data support a genetic contribution to the pathogenesis of IBD. Recently, numerous new genes have been identified as being involved in the genetic susceptibility to IBD: TNF-308A, CARD15 (NOD2), MIF-173, N-acetyltransferase 2 (NAT2), NKG2D (natural killer cell 2D), STAT6 (signal transducer and activator of transcription 6), CTLA-4 (cytotoxic T lymphocyte antigen-4), MICA-MICB (major histocompatibility complex A and B), HLA-DRB1, HLA class-II, IL-18, IL-4, MICA-A5, CD14, TLR4, Fas-670, p53 and NF-kappaB. The characterization of these novel genes has the potential to identify therapeutic agents and aid clinical assessment of phenotype and prognosis in patients with IBD (UC and CD).

Two new triterpenoid saponins, ardipusillosides IV and V (1 and 2, resp.), together with one known saponin, ardisiacrispin B (3), were isolated from the whole plants of Ardisia pusilla A. DC. Their structures were deduced by extensive spectral analysis and chemical evidences. Compound 1 contains a glycosylated glycerol residue which is a very rare structural feature among triterpenoid glycosides and has been so far found only in the genus Ardisia. All the saponins exhibited significant cytotoxicity against human glioblastoma U251MG cells, but did not affect the growth of primary cultured human astrocytes.

A I M : To e v a l u a t e t h e e f f i c a c y a n d s a f e t y o f gemcitabine-oxaliplatin (GEMOX) combined with huachansu (cinobufagin) injection treatment in patients with locally advanced or metastatic gallbladder carcinoma (GBC), and to assess the quality of life (QOL) of such patients.METHODS: Twenty-five patients with locally advanced or metastatic GBC were treated with intravenous gemcitabine (1000 mg/m 2 ) over 30 min on days 1 and 8, 2 h infusion of oxaliplatin (120 mg/m 2 ) on day 1, and 2-3 h infusion of huachansu (20 mL/m 2 ) on days -3-11, every 3-4 wk.Treatment was continued until occurrence of unacceptable toxicity or disease progression.QOL of patients was assessed by the EORTC QLQ-C30 at baseline, at the end of the first, third and sixth chemotherapy cycles, and 1 mo after the treatment.RESULTS: Among the 25 patients with a median age of 64 years (range 42-78 years), 23 were evaluable in the study.A total of 137 cycles of therapy were performed and the median cycle was 5 (range 1-8) per patient.Out of the 23 patients whose response could be evaluated, 8 partial responses (PR) were observed (34.8%), while 7 patients (30.4%) demonstrated a stable disease (SD).The disease control rate was 65.2%.Progression of cancer was observed in 8 (34.8%) patients.The median progression-free and overall survival time was 5.8 mo (95% CI: 4.5-7.1 mo) and 10.5 mo, respectively.The therapy was well tolerated, with moderate myelosuppression as the main toxicity.Anemia grade 2 was seen in 16.0%, neutropenia grade 3 in 8.0% and thrombocytopenia grade 3 in 24.0% of patients, respectively.Nonhematologic toxicity ranged from mild to moderate.No death occurred due to toxicity.The QOL of patients was improved after chemotherapy, and the scores of QOL were increased by 10 to 20 points.CONCLUSION: GEMOX combined with huachansu (cinobufagin) injection is well tolerated, effective, thus improving the QOL of patients with advanced GBC.