Background: Intrahepatic cholangiocarcinoma (ICC) is a highly desmoplastic tumor with poor prognosis even after curative resection. We investigated the associations between the composition of the ICC stroma and immune cell infiltration and aimed to develop a stromal-immune signature to predict prognosis in surgically treated ICC. Patients and methods: We recruited 359 ICC patients and performed immunohistochemistry to detect α-smooth muscle actin (α-SMA), CD3, CD4, CD8, Foxp3, CD68, and CD66b. Aniline was used to stain collagen deposition. Survival analyses were performed to detect prognostic values of these markers. Recursive partitioning for a discrete-time survival tree was applied to define a stromal-immune signature with distinct prognostic value. We delineated an integrated stromal-immune signature based on immune cell subpopulations and stromal composition to distinguish subgroups with different recurrence-free survival (RFS) and overall survival (OS) time. Results: We defined four major patterns of ICC stroma composition according to the distributions of α-SMA and collagen: dormant (α-SMAlow/collagenhigh), fibrogenic (α-SMAhigh/collagenhigh), inert (α-SMAlow/collagenlow), and fibrolytic (α-SMAhigh/collagenlow). The stroma types were characterized by distinct patterns of infiltration by immune cells. We divided patients into six classes. Class I, characterized by high CD8 expression and dormant stroma, displayed the longest RFS and OS, whereas Class VI, characterized by low CD8 expression and high CD66b expression, displayed the shortest RFS and OS. The integrated stromal-immune signature was consolidated in a validation cohort. Conclusion We developed and validated a stromal-immune signature to predict prognosis in surgically treated ICC. These findings provide new insights into the stromal-immune response to ICC.

Unlike healthy, non-transformed cells, the proteostasis network of cancer cells is taxed to produce proteins involved in tumor development. Cancer cells have a higher dependency on molecular chaperones to maintain proteostasis. The chaperonin T-complex protein ring complex (TRiC) contains eight paralogous subunits (CCT1-8), and assists the folding of as many as 10% of cytosolic proteome. TRiC is essential for the progression of some cancers, but the roles of TRiC subunits in osteosarcoma remain to be explored. Here, we show that CCT4/TRiC is significantly correlated in human osteosarcoma, and plays a critical role in osteosarcoma cell survival. We identify a compound anticarin-β that can specifically bind to and inhibit CCT4. Anticarin-β shows higher selectivity in cancer cells than in normal cells. Mechanistically, anticarin-β potently impedes CCT4-mediated STAT3 maturation. Anticarin-β displays remarkable antitumor efficacy in orthotopic and patient-derived xenograft models of osteosarcoma. Collectively, our data uncover a key role of CCT4 in osteosarcoma, and propose a promising treatment strategy for osteosarcoma by disrupting CCT4 and proteostasis.

OBJECTIVE: To establish the in vivo traceable acute myeloid leukemia mice model with Luciferase-Expressing KG1a Cells. METHODS: KG1a cells with stable luciferase gene expression (called as KG1a-Luc cells) were constructed by lentivirus transfection, then sifted out by puromycin. Eighteen male NOD-SCID-IL2rg RESULTS: KG1a cells expressing luciferase stably were successfully obtained. The tumor luminescence wildly spread at day 17 captured by in vivo imaging. The KG1a-Luc tumor cells could be detected in the peripheral blood of the mice, with the average percentage of (16.27±6.66)%. The morphology and pathology result showed that KG1a-Luc cells infiltrate was detected in bone marrow, spleens and livers. The survival time of the KG1a-Luc mice was notably shorter as compared with those in the control group, the median survival time was 30.5 days (95%CI: 0.008-0.260). CONCLUSION The acute myeloid leukemia NOD-SCID-IL2rg
Animals ; Disease Models, Animal ; Interleukin Receptor Common gamma Subunit ; Leukemia, Myeloid, Acute ; Luciferases/genetics* ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID

【Objective】To assess the difference in volume load between different blood pressure control levels in outpatients with hypertension. To investigate the effect of clinical factors on volume load in outpatients with hypertension. To analyze the clinical baseline characteristics and use of antihypertensive drugs in outpatients with hypertension. 【Methods】 A total of 514 outpatients with hypertension were included from July to November 2017. Clinical indicators including gender，age，height，weight，years of hypertension，blood pressure levels，treatment options for hypertension and comorbidities of these patients were recorded. The body volume load was evaluated by detecting the ratio of extracellular fluid（ECW）to total body water（TBW）using a Bioimpedance Analyzer. Whole body ECW/TBW≥0.39 was defined as high volume load. The effects of clinical factors on the volume load of hypertensive patients and whether there was a difference in volume load between different antihypertensive therapy were analyzed. 【Results】The blood pressure compliance rate of outpatients with hypertension was 15.37% ，which was still very low. Male patients had lower blood pressure compliance rate than female patients. Blood pressure was more difficult to control in patients with older age ，higher body mass index （BMI），higher waist-to-hip ratio（WHR）or longer duration of hypertension. The higher the blood pressure grading，the higher the proportion of combination medication. Diuretics were still the most widely used antihypertensive drugs. Age ，gender and different hypertension grades were the main factors affecting the volume load of hypertensive patients. Volume load was higher in female，older or higher systolic blood pressure（SBP）patients. Among them，age was the most important factor affecting the volume load of hypertensive patients.【Conclusion】The blood pressure compliance rate of outpatients with hypertension was still low. Effectively reducing the volume load was one of the important means to control blood pressure，which was more important in female，older or higher SBP patients.

PURPOSE: An older female predominance has been reported among chronic cough patients in Western countries, which is considered to be associated with a higher cough sensitivity in females. However, the characteristics of Chinese chronic cough patients remain unclear. This study aimed to explore the age and sex distribution as well as their relationship with cough reflex sensitivity to capsaicin in Chinese chronic cough patients. METHODS: We analyzed the demographic features of 1,882 consecutive chronic cough patients who attended our cough clinic in Guangzhou, China. Cough sensitivity to capsaicin, which was defined as the lowest concentration of capsaicin causing 5 coughs or more (C5), was measured in 539 of the 1,882 patients and 68 healthy volunteers. RESULTS: The mean age of the patients was 43.0 ± 13.7 years and patients aged <50 years accounted for more than two-thirds of the study population. Around 87% of the patients were never-smokers. The proportion of females (51.5%) was almost equal to that of males (48.5%). The pattern of the age and sex distribution was consistently reflected within most common causes of chronic cough, while a female predominance was shown in patients with cough-variant asthma and patients aged ≥50 years. Female patients had higher cough sensitivity to capsaicin than male patients (log C5: 1.58 ± 0.84 vs. 2.04 ± 0.84 μmol/L, P = 0.001), and patients aged ≥50 years had higher cough sensitivity to capsaicin than patients aged <50 years. CONCLUSIONS: In China, patients with chronic cough have a roughly equal sex distribution and a middle-aged predominance, irrespective of a higher cough sensitivity to capsaicin in females and older patients. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02591550
Age Distribution ; Asian Continental Ancestry Group ; Asthma ; Capsaicin ; China ; Cough ; Female ; Healthy Volunteers ; Humans ; Male ; Reflex ; Sex Distribution

Objective To investigate the clinical effect of laparoscopic and open radical resection of hilar cholangiocarcinoma (Bismuth-I). Methods From January 2011 to January 2017, 34 patients (10 patients in the laparoscopic group and 24 patients in the open group) underwent radical resection of hilar cholangiocarcinoma. Their clinical data were analyzed retrospectively. Results All these patients underwent radical resection of hilar cholangiocarcinoma of type Bismuth-I successfully. The volume of blood loss was (179.50 ± 98.05) ml and duration of hospital stay was (11.80 ± 2.49) d of laparoscopic group which was lower than (261.25 ± 97.33) ml, (16.25 ± 3.35) d of open group (P < 0.05). The levels of Tbil of laparoscopic group at days after operation 1, 3, 5 d were (102.20 ± 45.49) mmol/L, (83.57 ± 30.66) mmol/L, (45.09 ± 18.41) mmol/L. The levels of Tbil of open group at days after operation 1, 3, 5 d were (148.17 ± 62.78) mmol/L, (121.60 ± 43.35) mmol/L, (80.59 ± 43.89) mmol/L. The difference was statistically significant (P < 0.05). And the laparoscopic group postoperative number of lymph nodes dissected (9.79 ± 3.05), postoperative complications (1 cases), positive margin (0 cases); and open group(9.30 ± 3.06), 3 cases, had no statistically significant differences were compared (P >0.05). Two groups of patients were followed up for 6~18 months. During the follow-up period, there was no recurrence or metastasis in the two groups. Conclusion Laparoscopic Bismuth-I radical resection of hilar bile duct carcinoma in lymph node dissection, postoperative complications, positive margin rate, prognosis and open type Bismuth-I hilar cholangiocarcinoma radical operation of similar effects, and more minimally invasive advantages. Therefore, laparoscopic radical resection for hilar cholangiocarcinoma of type Bismuth-I is safe and feasible.

[Objective]To explore the correlations between different indices of lipoprotein cholesterol and apolipopro-tein of coronary heart disease(CHD)patients,especially that between low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)and that between apolipoprotein A1(apoA1)and apolipoprotein B100 (apoB100),as well as the correlations between these indices,indices ratios and the severity of coronary artery lesion.[Methods]301 coronary heart disease patients hospitalized to accept percutaneous coronary intervention(PCI)in the Third Affiliated Hospital of Sun Yat-sen University during 2013-2014 were recruited in the study. Fasting serum lipid indices including triglycerides(TG),total cholesterol(TC),LDL-C,HDL-C,apoA1 and apoB100 were examined before surgery.Gensini score was calculated to evaluate the severity of coronary artery lesion. 153 patients whose Gensini score was less than 50 were assigned to Group A,while 148 patients with Gensini score greater than or equal 50 were distributed to Group B.[Results]Positive correlations were found between LDL-C and HDL-C(r=0.161,P=0.005), apoA1 and apoB100(r=0.358,P<0.001),apoB100 and LDL-C(r=0.487,P<0.001),apoA1 and LDL-C(r=0.178, P=0.002)by linear correlation analysis. No significant correlation was found between apoB100 and HDL-C. None of LDL-C,HDL-C,TC was correlated with Gensini score. However,LDL-C/HDL-C ratio was positively correlated with Gensini score(r=0.148,P=0.01). The results showed no significant correlations between apoB100,apoB100/apo A1 ratio and Gensini score but negative correlation between apoA1 and Gensini score(r=-0.129,P=0.025). The positive correlation between HDL-C/LDL-C ratio and Gensini score was still valid after multi-factors adjustment (β=5.071,P=0.018).[Conclusion]Of patients with coronary heart disease,there exist some correlations between LDL-C and HDL-C,apoA1 and apoB100,while the correlation between LDL-C and HDL-C is relatively weak.The LDL-C/HDL-C ratio,weakly positively correlated with the severity of coronary artery lesion,is a risk factor of coronary artery lesion,while the level of apoA1,negatively correlated with the severity of coronary artery lesion,could play a protective role.

Objective To investigate the effects of atorvastatin on C-reactive protein (CRP)induced Toll-Like receptor 4(TLR4)expression on CD14+ monocyte, and the production of proinflammatory cytokines tumor necrosis factor α (TNFα), interleukin-6 (IL-6), matrix metalloproteinases-9 (MMP-9), and to study the anti-inflammatory mechanisms of statins. Methods The monocytes were isolated from blood of healthy volunteers by the Ficoll density gradient and stimulated by CRP with different doses (5, 25, 50, 100 μg/ml)and different exposure time (6, 12, 24, 48 h). Cells were also incubated with atorvastatin of different doses (1.0, 2.5, 5.0, 7.5, 10.0 μmol/L)in the presence of CRP 50 μg/ml. The protein expression of TLR4 was measured by flow cytometry, mRNA expression of TLR4 and of myeloid differentiation protein (MD2)was detected by quantitative PCR. TNFα, IL-6, MMP-9 concentrations in supernatants of cultured medium were measured by ELISA.Results (1)Compared with the un-stimulated control group, enhanced TLR4 protein expression was already detected at a concentration of 5 μg/ml of CRP and increased in a dose-dependent manner (32.22±2.80)%, (49.94±5.58)%, (74.82±3.24)% and (90.82±2.88)% at 5, 25, 50 and 100 μg/ml CRP. (2)TLR4 protein expression on 50 μg/ml CRP stimulated cells also increased in a time-dependent manner (29.80±2.70)%, (47.44±4.41)%, (81.71±2.92)% and (50.57±3.34)% after 6 h, 12 h, 24 h, 48 h.(3)When monocytes were incubated with CRP 50 μg/ml and atorvastatin (1.0, 2.5, 5.0, 7.5, 10.0 μmol/L), protein expression [(68.17±1.71)%, (52.43±1.38)%, (27.72±4.55)%, (17.46±3.20)%, (9.99±2.81)%］and mRNA expression (82.72%, 67.34%, 48.16%, 30.88%, 13.85%)of TLR4 as well as mRNA expression of MD2 (81.78%, 71.04%, 47.85%, 27.06%, 18.30%)were reduced in a dose-dependent manner. (4)Level of TNFα, IL-6 and MMP-9 in supernatants was significantly reduced by atorvastatin (2.5 μmol/L)compared with control group(P<0.01). When monocyte incubated with CRP 50 μg/ml and atorvastatin 10.0 μmol/L, the level of TNFα, IL-6, MMP-9 decreased to (25.8±2.5)pg/ml, (128.2±14.7)pg/ml, (65.2±12.3)ng/ml, respectively.Conclusion CRP increased the protein expression of TLR4 on CD14+ monocyte in a dose-dependent and time-dependent manner. Atorvastatin can inhibit the signal transduction of TLR4 and reduce proinflammatory cytokines release induced by CRP on CD14+ monocyte, and this might be one of the anti-inflammatory mechanisms of atorvastatin.

Objective To identify epitope of neutrophil-activating protein(NAP) of Helicobacter pylori(Hp).Methods Using the mouse monoclonal antibodies against NAP as selective molecular and immunoscrcening phage-display random 7-peptides library.The positive clones were sequenced and analyzed.Phage clones were chosen to immunize mice and to evaluate the potential of phagotopes as effective vaccines.Results One mimotope(FAHLATQ)showed a good match with the NAP at 140-143 AA(AHLA)and the serum of mice induced by the phage clone clearly recognized NAP.Conclusion This study suggests thatthe antigenic epitope could be mapped through screening the phage-display peptide library with monoclonalantibody and a mimotopo of NAP providing an ahernative approach for the diagnosis and development of avaccine for Hp.

Objective To investigate the clinical features of unexpected sudden death (SUD) clustered in families in Yunnan province.Methods This retrospective study analyzed the clinical features of SUD occurred between July to September 2005 in 7 families in Yunnan province.Results All 16 SUD patients shared common clinical features such as fatigue and repeated syncope and one group of SUD patients (n = 8 from 4 families) presented with the gastric intestinal tract manifestations including nausea, vomiting, abdominal pain and diarrhea with suspected dietary history and abnormal laboratory enzyme findings (GOT/ GPT,CK/CKMB,LDH/LDH1 etc.).In SUD patients without gastric intestinal tract manifestations (n = 8 from 3 families), there were no clear symptoms before death and repeated ventricular tachycardia and ventricular fibrillation were recorded in one survivor.There was no clear evidence for the involvements of hereditary and infectious factors for observed SUD.conclusion The reason for the unexpected sudden death clustered in 7 families in Yunnan remains unclear.Repeated syncope and fatigue served as the common clinical features in the presence or absence of gastric intestinal tract manifestations in all SUD cases.Further studies are needed to clarify the pathology and detailed clinical manifestations of SUD occurred in this area.