Search

Phytochemical analysis of Chloranthus henryi var. hupehensis roots led to the identification of a new eudesmane sesquiterpenoid dimer, 18 new sesquiterpenoids, and three known sesquiterpenoids. Among the isolates, 1 was a rare sesquiterpenoid dimer that is assembled by a unique oxygen bridge (C 11 -O-C 8' ) of two highly rearranged eudesmane-type sesquiterpenes with the undescribed C 16 carbon framework. (+)-2 and (-)-2 were a pair of new skeleton dinorsesquiterpenoids with a remarkable 6/6/5 tricyclic ring framework including one γ-lactone ring and the bicyclo[3.3.1]nonane core. Their structures were elucidated using spectroscopic data, single-crystal X-ray diffraction analysis, and quantum chemical computations. In the LPS-induced BV-2 microglial cell model, 17 suppressed IL-1β and TNF-α expression with EC 50 values of 6.81 and 2.76 µM, respectively, indicating its excellent efficacy in inhibiting inflammatory factors production in a dose dependent manner and without cytotoxicity. In subsequent mechanism studies, compounds 3, 16, and 17 could reduce IL-1β and TNF-α production by inhibiting IKBα/p65 pathway activation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

In this study, 16 new compounds, six bibenzyls (1-6) and 10 naphthalenes (7-13), including three pairs of naphthalene enantiomers and three known compounds (14-16), were isolated from Dendrobium chrysanthum. Structurally, compounds 1-5 are previously undescribed dimeric bibenzyls, uniquely linked by unusual carbon bonds. The structures of the compounds were determined using spectroscopy and X-ray crystallography. The screening results indicated that 1, 2, and 5 showed remarkable lipid-lowering activities in FFA-induced HepG2 cells, with EC 50 values ranging from 3.13 to 6.57 μM. Moreover, 1, 2, and 5 significantly decreased both the mRNA and protein levels of the target SREBP-1c, and 5 also reduced PPARα mRNA and protein levels. Therefore, 1, 2, and 5 are potential drugs against hepatic steatosis by targeting PPARα or SREBP-1c., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Eleven new amides, four racemic pairs of (±)-chlorahupetamides A, B, D, E (1, 2, 4, 5) and chlorahupetamides C, F, G (3, 6, 7), have been isolated from Chloranthus henryi var. hupehensis. Compounds 1-3 are the first naturally occurring dimers via an unprecedented [2 + 2] cycloaddition derived from two dissimilar cinnamic acid amides, while compounds 4 and 5 represent the first examples of lignanamides in Chloranthus; together with two new hydroxycinnamic acid amide monomers (6-7), these compounds were obtained. Their structures were characterized by nuclear magnetic resonance (NMR), electronic circular dichroism (ECD), and X-ray diffraction analysis. Meanwhile, an LPS-induced BV-2 cell inflammatory model was used to determine the potential anti-inflammatory activity of all the isolated compounds. Intriguingly, compound -1 treatment showed a much greater inhibition of TNF-α expression with an EC 50 value of 1.80 µM, while compound + 1 had more advantages in reducing IL-1β expression with an EC 50 value of 19.93 µM. Moreover, compounds + 1 and -1 could significantly suppress inflammation and inhibit the Akt signaling pathway by decreasing the phosphorylated protein levels of Akt., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

(+)- and (-)-Chlorahupetenes A ( 1a and 1b ), B ( 2a and 2b ), C ( 3a and 3b ), and D ( 4a and 4b ), four unique enantiomeric pairs of eudesmane-type sesquiterpenoid dimers with two new carbon skeletons, were isolated from the aerial parts of Chloranthus henryi var. hupehensis . Compounds 1 and 2 possess an unprecedented 6/6/5/6/6 pentacyclic carbon skeleton with a new dimerization pattern of two eudesmane-type sesquiterpenoids. Compounds 3 and 4 , which are fused with two eudesmane-type sesquiterpenoids via an unprecedented five-membered O -heterocyclic ring, represent a new 6/6/5/5/6/6/5 heptacyclic ring system. The structures of the compounds were determined through spectroscopic data and X-ray crystallography. Compounds 1a - 3b significantly inhibited NO production with IC 50 values ranging from 9.62 to 12.91 μM. Moreover, compounds 1b and 3a suppressed the production of a proinflammatory mediator (TNF-α) and enzyme expression (iNOS) at the mRNA level.

Objective: To analyze the characteristics of vocal fold vibration of normal people and patients with spasmodic dysphonia (SD) using laryngeal high speed videoendoscopy combined with glottal area wave analysis. Methods: This prospective study examined twenty healthy subjects (10 males and 10 females), 12 patients with adductor spasmodic dysphonia(AdSD) (2 males,10 females)as AdSD group and 2 patients with abductor spasmodic dysphonia(AbSD) (2 males) as AbSD group. Twelve of healthy subjects (2 males,10 females) were selected as control group according to AdSD group gender match. All the subjects were recruited from the Department of Voice, Zhongshan Hospital, Xiamen University from October 2019 to December 2020. All subjects underwent laryngeal high speed videoendoscopy and 10 vibration periods were selected from each recording and were used to quantitatively analyze the change of glottal area and vocal fold vibration parameters (Speed Quotient (SQ), Open Quotient (OQ) and Close Quotient (CQ)). Results: 1. There were statistically significant differences in SQ, CQ and OQ between males and females in the healthy subjects ( t =12.28, 5.59, 5.59, P <0.05). The change of the glottal area during each vibration period in healthy subjects was relatively stable(0.19-0.42). 2. The change of the glottal area during each vibration period in AdSD subjects had larger fluctuations, with the glottal area change index fluctuating in the range of 0.31 to 0.62. The SQ value of the AdSD group was significantly lower than that of control group ( t =4.246, P <0.05). There were no significant differences in OQ and CQ between AdSD group and normal group ( t =1.064, 1.332, P >0.05); The SQ value of the AbSD group tended to increase compared to normal group. Conclusions: Laryngeal high speed videoendoscopy combined with glottal area wave analysis has a certain reference value in the studying the vibration characteristics of SD patients. SQ has good specificity.

Interleukin‑6 (IL‑6) is involved in various biological responses, including tumor progression, metastasis and chemoresistance. However, the role and molecular mechanism of IL‑6 in the treatment of sorafenib in liver cancer remain unclear. In the present study, through western blot analysis, Transwell assay, flow cytometric assay, ELISA analysis and immunohistochemistry it was revealed that sorafenib promoted metastasis and induced epithelial‑mesenchymal transition (EMT) in liver cancer cells in vitro and in vivo, and significantly increased IL‑6 expression. Endogenous or exogenous IL‑6 affected metastasis and EMT progression in liver cancer cells through Janus kinase 2/signal transducer and activator of transcription 3 (STAT3) signaling. Knocked out IL‑6 markedly attenuated the pro‑metastasis effect of sorafenib and increased the susceptibility of liver cancer cells to it. In conclusion, the present results indicated that IL‑6/STAT3 signaling may be a novel therapeutic strategy for liver cancer.

Objective: To explore the dynamic changes of three-dimensional morphology of laryngeal soft tissue and its clinical value in the unilateral vocal fold paralysis (UVFP) patients through dynamic CT scanning during the process from inspiration to phonation. Methods: From October 2017 to July 2019, a retrospective study was performed in 18 patients with UVFP (10 males and 8 females with the range of age from 29 to 75 years old) and 10 normal subjects (5 males and 5 females with the range of age from 25 to 58 years old) in Department of Voice-Otolaryngology Head and Neck Surgery, Section Two, Zhongshan Hospital Xiamen University. The laryngeal dynamic computed tomography (CT) of cine mode was performed. Ten dynamic sequence images of vocal folds movements were obtained during the process from inspiration to phonation. Based on the dynamic changes of glottic area and the displacement of cricoid cartilage. The above dynamic sequence images were divided into inspiratory phase and phonation phase as well as open phase and closed phase. The soft tissue parameters were measured respectively, including vocal folds length, width, thickness and subglottal convergence angle. Independent-sample t test was used to analyze between UVFP group and control group. Results: During the process from inspiration to phonation, the morphology of vocal folds in control group was relatively stable at inspiratory phase and closed phase in phonation. When open phase and closed phase of phonation were switching, the morphology of vocal folds changed obviously. The length of vocal folds became longer (1.19±0.10) mm, the width became wider (2.19±0.17) mm, the thickness became thinner (2.66±0.56) mm, and the subglottal convergence angle decreased (31.45±4.78)°. Compared with the controll group, in the open phase, the thickness and width of the vocal fold on affected side in the UVFP group were thinner ( t= 10.25, P< 0.001) and wider ( t= 5.25, P< 0.001).While in the closed phase, the subglottal convergence angle was larger ( t= 4.41, P= 0.001).The width of the healthy side vocal fold in the UVFP was wider ( t= 2.54, P= 0.026) than that in the control group. The differences in other parameters were not statistically significant. Conclusions: Dynamic laryngeal CT scanning provides a simple and non-invasive method for the objective and quantitative measurement of the dynamic changes of laryngeal morphology from inspiration to phonation. Compared with the control group, the characteristic dynamic changes among UVFP were observed during this particular process, which included changes of subglottal convergence angle and thickness of vocal muscle due to denervation. In addition, in UVFP group, the width of the vocal fold healthy side in the closed phase may be used to assess its compensatory function.

One new sesqui-neolignan compound, namely, sesqui-illisimonan A (1), one new neolignan, illisimonan A (2), and one new phenylpropanoid compound, illisimoid A (3) were isolated from the fruits of Illicium simonsii Maxim. The structures and absolute configurations of these compounds were determined by extensive spectroscopic, including NMR, circular dichroism and calculated electronic circular dichroism methods. The antioxidant activities of compounds 1-3 were also evaluated. Vitamin E was selected as the positive control (IC 50  = 49.73 ± 0.88 μM). Compounds 1 and 2 exhibited in vitro antioxidant activity with an IC 50 value of 55.76 ± 1.30 and 59.36 ± 0.50 μM, respectively. However, the compound 3 didn't show obvious antioxidant activity.

Objective: To discuss the clinical effect of small dose of botulinum toxin injection in cricothyroid muscle and thyroarytenoid muscle on patients with incomplete bilateral recurrent laryngeal nerve paresis. Methods: Six patients were selected with Ⅰor Ⅱ or Ⅲ degree of dyspnea diagnosed as bilateral recurrent laryngeal nerve injury by laryngeal electromyography, and small dose of botulinum toxin injection was performed in cricothyroid muscle and thyroarytenoid muscle as a treatment. Degree of dyspnea was assessed one month before and after the treatment, and the stroboscopic laryngoscope results, acoustic parameters and CT image of the patients were collected in the 6 patients. The relevant parameters were also collected one month before and after treatment, including the degree of dyspnea, stroboscopic laryngoscope results, acoustic parameters and CT image of the patients. The angle between bilateral vocal cords in stroboscopy at full inspiratory was calculated, acoustic parameters (F0, jitter, shimmer) were analysed, and vocal length, width and the vocal region were measured. Then, the paired t test was performed for statistical analysis between before and after one month injection, the one way analysis of variance was performed among vocal parameters in CT image. Result: Botulinum toxin injection was successfully completed in the 6 patients, followed without any serious complications. The degree of dyspnea was alleviated to some extent after treatment in all 6 patients; the angle between bilateral vocal cords at the end of a deep inspiration was significantly increased ( t =2.44, P <0.05) after the treatment. The changes of F0 and jitter between before and after treatment were not statistically significant ( t =0.72, t =1.42, P >0.05). Shimmer was significantly decreased after treatment ( t =2.61, P <0.05). Vocal fold length, width and vocal region increased with F0, there was a statistically significant difference between different F0 before injection, and there was no statistically significant difference between different F0 after injection. The follow-up time was respectively seven months, 1 year, 1 year, 18 months, 22 months and 2 years respectively. Conclusion: Small dose of botulinum toxin injection in bilateral cricothyroid muscles and thyroarytenoid muscles can relieve the dyspnea caused by bilateral vocal cords paresis to some extent, accompanied without serious complications, despite the sound quality was slightly worse.

Objectives: To evaluate the therapeutic effect on the voice recovery of patients with vocal cord polyps undergoing the microsurgery of preoperative voice therapy., Methods: Twenty-six patients diagnosed with unilateral vocal cord polyp under stroboscope, who needed to undergo vocal cord loss resection under supportive laryngoscope, were randomly divided into control group (non-voice training) and treatment group (voice training), with each group of 13 patients. Patients in control group were just treated with surgical operation. Apart from surgical treatment, patients in treatment group received 6 hours intensive vocal therapy one week before the surgery. The therapy courses consist of the propaganda and education of voice care, postoperative vocal instruction and the patients' self-training under the guidance of voice therapists. The acoustic parameters (irregularity, breathiness, grade, jitter and shimmer) of the same patient were collected 24 to 48 hours before the surgery and 14 days after the surgery with Ling WAVES. The results were analyzed with SPSS 19.0., Results: The differences of all the five preoperative voice parameters between control group and treatment group are not significant; but postoperative breathiness and jitter in treatment group were significantly lower than that in control group, while the differences of irregularity, overall severity and shimmer were not significant between control group and treatment group. In control group, breathiness and jitter were significantly improved after surgery, while the differences of irregularity, breathiness and shimmer were not significant between preoperation and postoperation. In treatment group, all the five voice parameters were significantly improved after surgery. According to the laryngostroboscopic examination, the vocal fold polyps were excised completely in both groups., Conclusions: Preoperative voice therapy contributes to the recovery of voice quality of the patients with vocal cord polyps. Combined intervention of surgery and voice therapy is an effective method to treat the patients with vocal cord polyps., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Purpose: To determine the exact effect of Interleukin-6 (IL-6) on tumor cell proliferation, apoptosis, invasion, and anti-cancer therapy in hepatocellular carcinoma (HCC)., Experimental Design: IL-6 was disrupted by transcription activator-like effector nucleases (TALEN) in HCCLM3 cells, and was used to evaluate the role of IL-6 on tumor cell proliferation, apoptosis, invasion and key signaling pathways involved in sorafenib and/or IFNα therapy., Results: IL-6 has no direct effect on cell proliferation and invasion but promotes cell apoptosis and up-regulate IL-33 and VEGF-A expression. IL-6 could attenuate the anti-proliferation effect by sorafenib and combination therapy but facilitate the pro-apoptosis of the combination therapy and augment the pro-invasive effect induced by single treatment. IL-6 could down-regulate p-STAT3, however up-regulate the p-MEK/p-ERK and NF-kB/iNOS expression, and it also facilitated the promotion on p-JAK2 and p-MEK/p-ERK by either sorafenib or IFN-α. in vivo study, IL-6 significantly promotes tumor growth. The combination treatment showed the highest inhibition on tumor growth which is derived from HCCLM3-IL6(-) cells., Conclusions: IL-6 has no direct effect on cell proliferation and invasion but promotes tumor cell apoptosis in vitro study. Sorafenib and combination therapies are suitable for HCC cells with low or no IL-6 expression confirmed in vivo study., Competing Interests: CONFLICTS OF INTEREST All authors declare no conflicts of interest.

A new neolignan, (-)-(7R,8S,7'E)-3',4-dihydroxy-3-methoxy-8,4'-oxyneoligna-7'-ene-7,9,9'-triol(1), and seven known compounds, 9-(tetrahydropyran-2-yl)-nona-trans,trans-2,8-diene-4,6-diyn-1-ol (2), 9-(tetrahydropyran-2-yl)-trans-non-8-ene-4,6-diyn-1-ol (3), lobetyol (4), lobetyolin (5),dehydrodieoniferyl alcohol (6), 5-hydroxymethylfurfural (7), and 4, 4'-dihydroxy-3, 3'-dimethoxy-trans-stilbene (8), were isolated from the H2O extract of Codonopsis pilosula. The structures of 1-8 were elucidated by spectroscopic methods including NMR, HR-ESI-MS, and CD. In addition, compounds 2 and 3 were isolated from the genus Codonopsis for the first time., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To compare the acoustic signal, mucosal wave and aerodynamic parameter (phonation threshold pressure, PTP) under different sub-glottal pressure (SGP) on the excised canine models with different extent of glottal incompetence. Methods: Perturbation measures and nonlinear dynamic measures were applied to analyze the acoustic signal (jitter, shimmer), mucosal wave [frequency(F), amplitude(A), phase(P)] and PTP from our study including 11 excised canine larynges with different extent of glottal incompetence (0 mm, 1 mm, 2 mm, 3 mm, n =11, respectively) under 1-4 kPa sub-glottal pressure. Results: There were significant differences between different groups in jitter, shimmer, amplitude, frequency and PTP under various SGPs and extent of glottal incompetence (all P <0.05), inversely, there was no significant difference in P between groups ( P >0.05). Jitter and shimmer changed obviously when the SGP increased to 3 kPa in the control group and GI 1 mm group. Jitter and shimmer changed obviously when the SGP increased to 2 kPa in the GI 2 mm and 3 mm groups. The F and A of mucosal wave increased with increasing SGP, decreased with increasing GI, and the P changed irregularly. There was statistically significant difference of PTP between different GI groups. Conclusions: The SGP and the extent of GI had obvious affection on the the acoustic signal, mucosal wave and aerodynamic parameters.

Objective: This paper seeks to identify useful and reliable indicators for predicting the occurrence of difficult laryngeal exposure(DLE) in microlaryngosurgery. Method: Sixty-two patients were given physical examinations,including 4 general parameters:age,sex,BMI,and MMI,and 14 physical measurement parameters(TA,UIA,LIA,IG,LIMD,MA,MCD,MH,MDI,HMD,TMD,SMD,TMA,THUIA).Univariate analysis,stepwise regression analysis and ROC curve analysis were employed to identify parameters with the potential to predict DLE. Result: We found sex( P <0.05) showed significant correlation with the laryngeal exposure score( P <0.05).We also found LIMD,MA,TMA and THUIA to be reliable DLE predictors.The cutoff values for predicting DLE were LIMD>4.53cm,MA>115.5°,TMA>99.2°,and THUIA>152.6°. Conclusion: X-ray measurement predictors of LIMD,MA,TMA,and THUIA before operation are important for the prediction of DLE.Sex is also the reliable DLE predictor.Males were prone to DLE., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: Analysis of the sustained vowels of acoustic parameters in arytenoid dislocation and vocal fold paralysis. To investigate their acoustic characteristics and evaluate the role of this acoustic analysis method in differentiating arytenoid dislocation from vocal fold paralysis. Method: Thirty-three cases with unilateral vocal cord movement disorders were collected.All cases were divided into arytenoid dislocation group and vocal fold paralysis group through the laryngeal electromyography. Each group was further devided into male group and female group. The voice signals of sustained vowel of /a/ were measured using the software MDVP and obtain the acoustic parameters(Jitter, Shimmer, SPI and Fo). The acoustic characteristics between the two groups were observed and compared. Results were analyzed using Rank sum test for group design. Result: There were significant differences in Fo between arytenoid dislocation group and vocal fold paralysis group in both male and female group（ P <0.05）. And mean rank order of Fo in arytenoid dislocation group was greater than vocal fold paralysis group. There were no significant differences in jitter,shimmer and SPI between arytenoid dislocation group and vocal fold paralysis group（ P >0.05）. Conclusion: Mean rank order of Fo in arytenoid dislocation group is greater than vocal fold paralysis group in both the male and the female group. Fo is of value in differentiating arytenoid dislocation from vocal fold paralysis. This provides a theoretical basis for the acoustic analysis method to identify these two diseases., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Background: This study aimed to explore the responses to the interleukin-6 (IL-6)/soluble interleukin-6 receptor (sIL-6R) complex in peritumoral endothelial cells (PECs) and tumor endothelial cells (TECs), as well as determine the signaling pathways in the angiogenesis of hepatocellular carcinoma (HCC)., Methods: The expression of IL-6, IL-6R, gp130, CD68, HIF-1α, and microvessel density (MVD) were assessed with an orthotopic xenograft model in nude mice. ECs were incubated under hypoxic conditions to detect IL-6 and gp130. The proliferation of PECs and TECs in the presence of IL-6 and sIL-6R, as well as the expression of gp130, JAK2/STAT3, PI3K/AKT in endothelial cells were measured., Results: Peritumoral IL-6, IL-6R, gp130, CD68, and HIF-1α expression, as well as MVD, gradually increased during tumor growth. Hypoxia could directly induce IL-6 expression, but not gp130 in PECs. The co-culture of IL-6/sIL-6R induced much higher PEC proliferation and gp130 expression, as well as the elevated phosphorylation of JAK2 and STAT3, however not the phosphorylation of PI3K and AKT., Conclusions: PECs exhibited higher proliferation in response to IL-6/sIL-6R co-treatment compared with TECs in HCC via the up-regulation of gp130 /JAK2/STAT3. PEC and its associated peritumoral angiogenesis microenvironment may be a potential novel target for anti-angiogenic treatment.

Purpose: To determined Neuropilin-1 (NRP-1) and vascular endothelial growth factor receptor 2 (VEGFR-2) expression in the tumoral and peritumoral tissues of 214 treatment-naïve HCC patients and its correlation with overall survival (OS) and time to recurrence (TTR)., Experimental Design: NRP-1 and VEGFR-2 expression were examined by tissue microarray and peritumoral hypoxia by pimonidazole staining and angiogenesis by microvessel density (MVD). OS and TTR were evaluated by Kaplan-Meier analysis and log-rank test., Results: Peritumoral NRP-1 and VEGFR-2 expression were significantly higher than that of the tumoral tissue (p < 0.001 for both), and high peritumoral expression of both factors was negatively associated with tumor size (p < 0.001 for both). Patients with high peritumoral expression of both proteins had the longest median OS (>94.0 months) and TTR (>84.0 months). The multivariate Cox proportional hazards analysis revealed that patients with high peritumoral expression of both NRP-1 and VEGFR-2 were more than 4 times less likely to have recurrence (p = 0.004) and more than 10 times likely to survive (p < 0.001)., Conclusions: Peritumoral NRP-1 and VEGFR-2 expression is associated with prolonged TTR and extended OS of HCC patients and both may be useful as predictors of surgical outcome of HCC patients and explored as potential therapeutic targets.

Background: A better understanding of the molecular mechanisms in liver regeneration holds promise for exploring the new potential therapy for liver failure. The present study was to investigate the role of zinc finger and BTB domain-containing protein 20 (ZBTB20), a potential factor associated with liver regeneration, in a model of 70% hepatectomy in mice., Methods: Parameters for liver proliferation such as liver/body ratio and BrdU positivity were obtained via direct measurement and immunohistochemistry. The levels of zinc fingers and homeoboxes 2 (ZHX2), ZBTB20, alpha-fetoprotein (AFP) and glypican 3 (GPC3) transcripts in the regenerating liver tissue of a 70% hepatectomy rodent model were monitored by real-time PCR analysis at different time points. Knockdown of ZBTB20 was performed to characterize its regulatory function., Results: A negatively regulating relationship between ZHX2, ZBTB20 and AFP, GPC3 was revealed from 24 to 72 hours after 70% hepatectomy. ZBTB20 appears to negatively regulate AFP and GPC3 transcription since the knockdown of ZBTB20 promoted the proliferation of hepatocytes and the expression of AFP and GPC3., Conclusion: In addition to AFP, GPC3 and ZHX2, ZBTB20 is a new regulator in liver regeneration and the decrease of ZBTB20 expression following 70% hepatectomy promotes AFP and GPC3 expression.

Seven new sesquiterpenoid glycosides, consisting of one thapsan-type (1), two capnellane-type (2 and 3), four floridanolide sesquiterpene glycosides (4-7), and one new azulene-type sesquiterpene (8), along with six known sesquiterpenes (9-14) were isolated from the roots of Illicium henryi. The structures of 1-8 were elucidated by extensive spectroscopic analyses and chemical methods. The absolute configurations of 1, 2, and 8 were determined based on Rh2(OCOCF3)4-induced CD, Mo2(OAc)4-induced CD data, and calculated electronic circular dichroism (ECD), respectively. Compound 1 was found to exhibit weak neurotoxicant activity., (Georg Thieme Verlag KG Stuttgart · New York.)

Antiangiogenic therapy, specially sorafenib, has become the standard of care for patients with advanced hepatocellular carcinoma (HCC), however, the improvement in survival time is not satisfactory. Previous studies have found that, in some circumstances, antiangiogenic therapy promoted tumor metastasis and the mechanistic studies were mainly focus on cancer-cell-autonomous manners. In two experimental metastasis models with tail-vein injection with hepatoma cells and an orthotopic HCC mouse model, we found that pretreatment with two vascular endothelial growth factor receptor (VEGFR) inhibitors, sunitinib and sorafenib, facilitated tumor cell survival in blood stream and promoted lung metastasis from tumors that were subsequently incubated after drug discontinuation, indicating that host response joined into the pro-metastatic effects. An antibody microarray identified that interleukin (IL)-12b was decreased in the peripheral blood of the mice treated with the two VEGFR inhibitors. IL-12b suppression in macrophages and dendritic cells from host organs was found to play a crucial role in treatment-induced metastasis. Supplement with recombinant mouse IL-12b or restoration of IL-12b expression in the host by zoledronic acid, which was previously reported to enhance IL-12 expression in vitro and in vivo, alleviated the metastasis-promoting effects of sunitinib and sorafenib. These studies suggest that host response to VEGFR inhibitors facilitates HCC metastasis and restoration of IL-12b expression could translate into clinical benefits.

Objective: To investigate the comprehensive measures for improving radical resection rate and safety of Bismuth-Corlette type III hilar cholangiocarcinoma., Methods: The clinical data of 15 patients with Bismuth-Corlette type III hilar cholangiocarcinoma who performed radical resection from June 2009 to December 2011 was analyzed retrospectively. There were 11 male and 4 female patients, aged from 45 to 74 years (mean 59 years). The preoperative evaluation were conducted by using magnetic resonance cholangiopancreatography (MRCP), dual source spiral CT combined with IQQA-Liver CT Imaging Analysis System providing three-dimensional reconstruction of tumor, bile duct, hepatic artery and portal vein, which could help to chose the appropriate treatment modality. All patients were treated with selective hemi-hepatic vascular control of removal liver, hemi-hepatectomy combined with whole caudate lobe resection and regional lymphadenectomy. The merits of each evaluation methods and measures of surgical treatment were analyzed thoroughly., Results: The preoperative evaluation modalities including the dual source spiral CT combined with IQQA-Liver CT Imaging Analysis System could clearly show the involvement of bile duct, hepatic artery and portal vein invaded by the tumor. The satisfactory postoperative recovery could be achieved by the remnant liver volume of > 40% after the hemi-hepatectomy combined with whole caudate lobe resection through the selective preoperative biliary drainage and hemihepatic vascular control. The mean complication was transient aggravated liver dysfunction. There was no death reported during perioperative period in the group., Conclusions: The preoperative imaging evaluation modalities including MRCP, dual source spiral CT combined with IQQA-Liver CT Imaging Analysis System could do favor for the preoperative evaluation of invasion degree of hilar cholangiocarcinoma and the selection of appropriate surgical treatment. Hemi-hepatectomy combined with whole caudate lobe resection and regional lymphadenectomy could be an alternative management of Bismuth-Corlette type III hilar cholangiocarcinoma.

Background: Peritumoral liver tissue could play a potential role in hepatocellular carcinoma (HCC) progression and patient survival via angiogenesis- and lymphangiogensis-related factors. The prognostic role of these factors in hepatocytes and stromal cells in HCC patients after curative resection remains to be explored., Methods: Tumor tissue and surrounding peritumoral tissue were obtained from 145 resected HCC patients without lymph node metastasis (LNM) and 37 resected HCC patients with LNM. Tissue microarrays were constructed from duplicate cores of tumor tissue and surrounding peritumoral tissue from each resected specimen. Immunohistochemistry and real-time polymerase chain reaction were used to evaluate the expression of vascular endothelial growth factor-A (VEGF-A), VEGF-C, VEGF receptor-1(VEGFR-1), VEGFR-2, and VEGFR-3. Macrophage infiltration was determined by CD68 staining. Correlations between the expression of these factors and overall survival (OS) and time to recurrence (TTR) were studied., Results: The peritumoral expression of VEGF-A, VEGF-C, VEGFR-1, VEGFR-2, and VEGFR-3 were significantly higher than expression of these factors in tumors. VEGFR-1 was mostly located in peritumoral macrophages, while VEGF-C and VEGFR-3 were mostly located in peritumoral hepatocytes. HCC with high peritumoral co-expression of VEGF-C, VEGFR-1, and VEGFR-3 was associated with higher peritumoral distribution of macrophages (0.87%±0.26% versus 0.45%±0.20%), LNM (32.4% versus 12.0%), shorter TTR (10.2 months versus 34.5 months), and poor prognosis (19.4 months versus 49.3 months)., Conclusion: Expression of VEGF-C, VEGFR-1, and VEGFR-3 in peritumoral liver tissue is associated with a unique type of HCC that has a poorer outcome after hepatectomy.

Eleven prenylated C(6)-C(3) compounds, illihenryifunones A, B (1, 2), illihenryifunol A (3), illihenryipyranol A (4), illihenryiones A-G (5-11), and three known prenylated C(6)-C(3) compounds (12-14), were isolated from the roots of Illicium henryi. Structures of 1-11 were elucidated by spectroscopic methods including NMR, HRESIMS, and CD. The absolute configuration of the 11,12-diol moiety in 5 was determined by observing its induced circular dichroism after addition of Mo(2)(OAc)(4) in DMSO. The absolute configuration of C-11 in 4 was determined as S based on the Rh(2)(OCOCF(3))(4)-induced CD data; the absolute configuration of 3 was determined as R by comparison of its experimental and calculated electronic circular dichroism (ECD). The antioxidant activities of compounds 1-14 were also evaluated. Compound 4 exhibited strong antioxidant activity with an IC(50) value of 2.97±1.30 μM, whereas compounds 3 and 8 showed antioxidant activities with IC(50) values of 44.36±0.30 and 48.00±2.01 μM, respectively., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Interferon (IFN)-α is effective in inhibiting tumor growth and metastasis of hepatocellular carcinoma (HCC). However, the biologic mechanisms of IFN-α treatment in lung metastasis are not yet clear., Methods: The effect of IFN-α treatment was studied by using an orthotopic xenograft model and measuring tumor size and lung metastasis. Pretreatment with IFN-α before implantation of tumor was done to explore the effect of IFN-α on lung tissues. Cytokines and macrophages were measured by immunohistochemistry and/or PCR assay, using human origin or mouse origin primers to differentiate the sources. Circulating tumor cells (CTCs) were also assayed by flow cytometry., Results: IFN-α treatment did not decrease the number of CTCs (0.075% ± 0.020% versus 0.063%±0.018%, P = 0.574, IFN-α-treated versus control groups), but did decrease the number and size of lung metastasis (number: 1.75 ± 1.0 versus 28.0 ± 6.3, P = 0.008; size [pixels]: 116.8 ± 72.2 versus 5226.4 ± 1355.7, P = 0.020), and inhibited macrophage infiltration (0.20% ± 0.04% versus 1.36% ± 0.21%, P = 0.0058) and alteration of matrix metalloproteinase (MMP)-9 expression (mean integrated optical density (IOD): 5.1 ± 1.7 versus 21.9 ± 0.4, P<0.000) in the lung, which was independent of the primary tumor., Conclusion: IFN-α inhibited lung metastasis by directly modulating the lung microenvironment.

Objectives: To determine the distribution of macrophages (MΦ) in both xanthogranulomatous cholecystitis (XGC) and gallbladder carcinoma (GBC) and to analyze the association between XGC and GBC., Methods: From January 2009 to June 2011, 110 patients with gallbladder diseases, including 35 with GBC, 45 with XGC and 30 with chronic cholecystitis (CC), were enrolled. Immunohistochemistry stain and real-time polymerase chain reaction using oncogenes (BCL-2, c-Myc) and anti-oncogene genes (p53, p21) were performed, serum expressions of tumor marker (CA19-9, CA724 and CA242) were also conducted. MΦ were used to determine their potential role in the carcinogenesis of GBC., Results: BCL-2 and c-Myc expressions gradually increased among CC, XGC and GBC (P = 0.032 and P = 0.020, respectively); while p53 and p21 were similar in the three groups (P = 0.167 and P = 0.122, respectively). Serum BCL-2 and c-Myc were significantly correlated with their tissue levels; in terms of serum tumor markers, which gradually increased among CC, XGC and GBC, however, CA242 and CA724 were both negative in XGC but positive in GBC. Furthermore, gradually increasing MΦ counts were observed among CC, XGC and GBC groups; c-Myc and CA724 were independent predictors for the differentiation of XGC and GBC., Conclusions: XGC is an uncommon inflammatory condition distinct from CC and may be associated with the precancerous nature of GBC for its upregulated oncogenes and MΦ biology. c-Myc and CA724 were independent predictors for the differentiation of XGC and GBC., (© 2012 The Authors. Journal of Digestive Diseases © 2012 Chinese Medical Association Shanghai Branch, Chinese Society of Gastroenterology, Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine and Wiley Publishing Asia Pty Ltd.)

Background & Aims: Antiangiogenic agents can sometimes promote tumor invasiveness and metastasis, but little is known about the effects of the antiangiogenic drug sorafenib on progression of hepatocellular carcinoma (HCC)., Methods: Sorafenib was administered orally (30 mg · kg(-1) · day(-1)) to mice with orthotopic tumors grown from HCC-LM3, SMMC7721, or HepG2 cells. We analyzed survival times of mice, along with tumor growth, metastasis within liver and to lung, and induction of the epithelial-mesenchymal transition. Polymerase chain reaction arrays were used to determine the effects of sorafenib on gene expression patterns in HCC cells. We analyzed regulation of HIV-1 Tat interactive protein 2 (HTATIP2) by sorafenib and compared levels of this protein in tumor samples from 75 patients with HCC (21 who received sorafenib after resection and 54 who did not)., Results: Sorafenib promoted invasiveness and the metastatic potential of orthotopic tumors grown from SMMC7721 and HCC-LM3 cells but not from HepG2 cells. In gene expression analysis, HTATIP2 was down-regulated by sorafenib. HCC-LM3 cells that expressed small hairpin RNAs against HTATIP2 (knockdown) formed less invasive tumors in mice following administration of sorafenib than HCC-LM3 without HTATIP2 knockdown. Alternatively, HepG2 cells that expressed transgenic HTATIP2 formed more invasive tumors in mice following administration of sorafenib. Sorafenib induced the epithelial-mesenchymal transition in HCC cell lines, which was associated with expression of HTATIP2. Sorafenib regulated expression of HTATIP2 via Jun-activated kinase (JAK) and signal transducer and activator of transcription (STAT)3 signaling. Sorafenib therapy prolonged recurrence-free survival in patients who expressed lower levels of HTATIP2 compared with higher levels., Conclusions: Sorafenib promotes invasiveness and the metastatic potential of orthotopic tumors from HCC cells in mice, down-regulating expression of HTATIP2 via JAK-STAT3 signaling., (Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.)

Objective: To investigate the application and significance of aerodynamic parameters in voice function assessment., Methods: The phonatory aerodynamic system (PAS) was used to collect aerodynamic parameters from subjects with normal voice, vocal fold polyp, vocal fold cyst, and vocal fold immobility. Multivariate statistical analysis was used to compare measurements across groups., Results: Phonation threshold flow (PTF), mean flow rate (MFR), maximum phonation time (MPT), and glottal resistance (GR) in one hundred normal subjects were significantly affected by sex (P < 0.05), while phonation threshold pressure (PTP), subglottal pressure (SGP), and vocal efficiency (VE) were not (P > 0.05). PTP, PTF, MFR, SGP, and MPT were significantly different between normal voice and voice disorders (P < 0.01), and there were no significant differences among the three disorders (P > 0.05). Receiver operating characteristic (ROC) analysis found that PTP, PTF, SGP, MFR, MPT, and VE in one hundred thirteen voice dis orders had similar diagnostic utility (P < 0.01), with PTP exhibiting the highest area under the curve. The aerodynamic parameters of the three degrees of voice dysfunction due to vocal cord polyps were compared and found to have no significant differences (P > 0.05). PTP, PTF, MFR, SGP and MPT in forty one patients with vocal polyps were significantly different after surgical resection of vocal cord polyps (P < 0.01)., Conclusion: The aerodynamic parameters can objectively and effectively evaluate the variations of vocal function, and have good auxiliary diagnostic value.

Background: Postoperative interferon-α(IFN-α) treatment delays hepatocellular carcinoma(HCC) recurrence and prolongs patient survival, and may thus be an effective form of adjuvant therapy. However, clinical observations found that HCC recurs in some patients within 8 months of IFN-α treatment being discontinued. We investigated whether HCC regrowth appears after IFN-α is discontinued, whether re-initiated IFN-α is effective, and the underlying mechanisms of IFN-α treatment., Methods: The human HCC nude mouse model LCI-D20 was used to study the effects of IFN-α treatment, discontinued IFN-α treatment, and re-initiated IFN-α treatment on tumor growth. Tumor weight, microvessel density(MVD), serum vascular endothelial growth factor (VEGF), and tumor cell apoptosis were analyzed. Angiogenesis-related factors were studied using cDNA microarray in different tumor samples and confirmed using reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting assays. Finally, imatinib was added with re-initiated IFN-α treatment to improve efficacy., Results: IFN-α (1.5 × 107 U/kg/day for 20 days) suppressed HCC growth by 60.3% and decreased MVD by 52.2% compared with the control. However, tumor regrowth occurred after IFN-α was discontinued, and re-initiated IFN-α treatment was not effective for inhibiting tumor growth or reducing MVD compared with a saline-treated group. cDNA microarray showed VEGF was down-regulated while platelet-derived growth factor-A (PDGF-A) was up-regulated when IFN-α treatment was re-initiated. These findings were further confirmed with RT-PCR and Western blotting assay. The combination of imatinib with re-initiated IFN-α reduced HCC weight by 30.7% and decreased MVD by 31.1% compared with IFN-α treatment only (P=0.003 and 0.015, respectively)., Conclusion: Tumor regrowth occurred after IFN-α treatment was discontinued. Re-initiated IFN-α treatment was not effective and was associated with up-regulation of PDGF-A, while the VEGF remained suppressed. The combination of a PDGF-receptor inhibitor with IFN-α improved the effect of the re-initiated treatment.

Lycojaponicumins A-C (1-3), three trace alkaloids isolated from Lycopodium japonicum, represent a unique heterocyclic skeleton formed by the new linkage C4-C9. Notably, lycojaponicumins A and B (1 and 2) are the first examples of natural products possessing a 5/5/5/5/6 pentacyclic ring system with a 1-aza-7-oxabicyclo[2.2.1]heptane moiety. These structures were elucidated by spectroscopic methods and X-ray diffraction analysis. A plausible biogenetic pathway was proposed.

This work aimed to overcome chitosan (CS) membrane' drawbacks: mainly stiffness and hydrophobic surface by adding poly(vinyl alcohol) (PVA) and evaluate their biocompatibility. The chemical structure, crystalline and thermal properties were studied by FT-IR, XRD and DSC. The mechanical properties and wettability of CS/PVA membranes were studied by tensile test and static contact angle measurement. In vitro biocompatibility was also evaluated by MTS cytotoxicity assay and SEM examination. The results suggest that adding PVA into CS membrane could greatly improve CS membrane's flexibility and wettability. All the membranes prepared were biocompatible and have potential applications in GTR technology., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: The aim of this study was to evaluate the diagnostic and prognostic role of staging laparoscopy in gallbladder carcinoma (GBC)., Methods: From January 2007 through December 2010, 79 GBC patients without evidence of metastatic disease on preoperative imaging underwent staging laparoscopy. Peritoneal and liver metastases were assessed by a single surgeon in a systematic manner. Resection rate, safety, and survival analysis were compared between the laparoscopy group and no laparoscopy group., Results: Disseminated disease was detected in 27 patients and no further surgery was performed; the overall accuracy for detecting unresectable disease was 67.5% (27/40), with 39 (75%) and 27 (51.9%) receiving resection and curative resection. In 203 GBC patients undergoing laparotomy, 90 (44.3%) and 53 (26.1%) patients received resection and curative resection; therefore, the resection rate and curative resection rate were significantly much higher in the laparoscopy group (p < 0.000)., Conclusions: Staging laparoscopy in GBC is sensitive in detecting disseminated disease and increases the curative resection rate, shortens the recovery time, and has no negative implications on overall survival; therefore, we suggest the routine use of staging laparoscopy in patients with GBC without evidence of disseminated disease on preoperative imaging., (Copyright © 2012 S. Karger AG, Basel.)

Gallbladder cancer is a rare disease and it is associated with a poor clinical outcome and survival. A standard therapy for it has not been established yet. The aim of this study is to evaluate efficacy and safety of two modified ECF regimens in advanced gallbladder cancer patients. Clinical data of 38 patients with advanced gallbladder cancer treated with modified ECF regimen were reviewed retrospectively. Of them, 21 patients received an epirubicin, cisplatin, and 5-FU/LV combination therapy. Seventeen patients received a chemotherapy of epirubicin, cisplatin, and capecitabine. Partial response was achieved in fourteen (36.84%) patients with a median duration of 5 months (range, 3-13 months), while stable disease was achieved in eight patients (21.05%). The median time to progression was 4.0 months (95% CI, 3.62-4.58 months). And the median overall survival was 9.8 months (95% CI, 7.26-12.34 months). Responders demonstrated better survival than non-responders (median survival time: 16 vs. 6.9 months, P = 0.008). The median survival time for epirubicin-, cisplatin- and capecitabine-treated patients was 9.2 versus 8.9 months for epirubicin-, cisplatin- and 5-FU/LV-treated patients. There was no statistical difference between both treatment groups in terms of survival time (P = 0.769). Regimen-related toxicity resulted in at least one treatment delay or dosage reduction in 63.2 and 34.2% patients, respectively. There were no chemotherapy-related deaths during the study. Modified ECF regimen with epirubicin, cisplatin and 5-FU/LV or substituting capecitabine for 5-FU/LV is still a potentially effective therapeutic chemotherapy for patients with advanced gallbladder cancer, and toxicity was manageable. There was no remarkable difference in efficacy between the two regimens.

Vascular endothelial growth factor-targeted therapy is a promising treatment for hepatocellular carcinoma (HCC), but its clinical benefit is often accompanied by acquired resistance. In animal studies, antiplacental growth factor therapy is effective with less resistance. The role of placental growth factor (PlGF) in the progression of HCC is not clear. In our study, we used immunohistochemistry in tissue microarrays to investigate PlGF expression in tumor and peritumoral liver tissues from 105 patients with HCC. Intratumoral and peritumoral PlGF mRNA expression was analyzed in another cohort of 37 patients. Peritumoral PlGF expression was significantly higher than intratumoral PlGF expression (p < 0.001). Intratumoral PlGF expression was not associated with patients' overall survival (OS) or time to recurrence (TTR). However, peritumoral PlGF expression, which was associated with tumor size, presence of intrahepatic metastasis, TNM stage and Barcelona Clinic Liver Cancer stage, was an independent risk factor for OS (p = 0.026) and TTR (p = 0.041). The prognostic value of peritumoral PlGF expression was further validated in a validation cohort (n = 394). We inferred that the elevation of PlGF in peritumoral liver might be induced by hypoxia. We found that peritumoral PlGF expression was associated with hypoxia-inducible factor-1α (p = 0.017). PlGF expression was elevated in L02, a hepatic cell line, under hypoxic conditions in vitro. These findings indicate that high peritumoral PlGF expression is associated with tumor recurrence and survival after resection of HCC. PlGF could be a target of adjuvant therapy and deserves further investigations., (Copyright © 2010 UICC.)

Purpose: In recent years, anti-angiogenesis drugs have shown promising clinical effects against many tumors, particularly in combination with chemotherapy. Although the combination has become a standard of care for many tumors, the mechanisms of the chemosensitizing activity of anti-angiogenic drugs are not fully understood. Here, we sought to determine if anti-angiogenesis drug bevacizumab could enhance the chemosensitivity of HCC by inhibition of survivin., Methods: After treatment of human umbilical vein endothelial cells (HUVECs) and hepatocellular carcinoma (HCC) cell line PLC/PRF/5 (PLC) with bevacizumab or/and adriamycin, the direct effects were examined by survival assays, and the expression of Akt, Phospho-Akt and survivin were evaluated by western blot. Tumor growth was observed in a human HCC xenograft nude mouse model treated with different drugs, and the expression of PCNA, CD31 and survivin in tumor tissues were evaluated by means of immunohistochemistry., Results: Bevacizumab enhanced the chemosensitivity of HCC by inhibiting the VEGF-PI3 K/Akt-survivin signaling cascade in endothelial cells. The combination of bevacizumab with adriamycin therapy resulted in better outcomes compared with monotherapy in hepatocellular carcinoma xenografts; bevacizumab significantly inhibited tumor angiogenesis and growth. In addition, bevacizumab reduced survivin expression in tumor tissues, including tumor vascular endothelial cells in vivo, although it did not inhibit survivin expression in tumor cells in vitro., Conclusion: These results implicate the bevacizumab-increased efficacy of adriamycin via an inhibition of survivin expression in malignant cells as well as tumor vasculature cells, which provides other insights into the mechanism of enhanced efficacy by combination of VEGF blocker and chemotherapeutic agents.

Background: Antiangiogenesis is a promising therapy for advanced hepatocellular carcinoma (HCC), but the effects are difficult to be evaluated. Pazopanib (GW786034B) is a pan-vascular endothelial growth factor receptor inhibitor, the antitumor effects or antiangiogenic effects haven't been investigated in HCC., Methods: In vitro direct effects of pazopanib on human HCC cell lines and endothelial cells were evaluated. In vivo antitumor effects were evaluated in three xenograft nude mice models. In the subcutaneous HCCLM3 model, intratumoral blood perfusion was detected by contrast-enhanced ultrasonography (CEUS), and serial quantitative parameters were profiled from the time-intensity curves of ultrasonograms., Results: In vitro proliferation of various HCC cell lines were not inhibited by pazopanib. Pazopanib inhibited migration and invasion and induced apoptosis significantly in two HCC cell lines, HCCLM3 and PLC/PRF/5. Proliferation, migration, and tubule formation of human umbilical vein endothelial cells were inhibited by pazopanib in a dose-dependent manner. In vivo tumor growth was significantly inhibited by pazopanib in HCCLM3, HepG2, and PLC/PRF/5 xenograft models. Various intratumoral perfusion parameters changed over time, and the signal intensity was significantly impaired in the treated tumors before the treatment efficacy on tumor size could be observed. Mean transit time of the contrast media in hotspot areas of the tumors was reversely correlated with intratumoral microvessel density., Conclusions: Antitumor effects of pazopanib in HCC xenografts may owe to its antiangiogenic effects, and the in vivo antiangiogenic effects could be evaluated by quantitative CEUS.

Purpose: To elucidate the effect of IFN-α treatment on tumor growth and metastasis of hepatocellular carcinoma (HCC)., Methods: IFN-α administration was conducted in nude mice using an orthotopic implantation model of human HCC, and the key molecular markers in the IFN-α treatment was detected by immunohistochemistry staining and PCR array., Results: Up to 12 weeks of IFN-α treatment significantly suppressed tumor growth of HCC, but relatively increased the number of circulating tumor cells, which might be due to the enhanced tumor hypoxia as well as up-regulation of metastasis-related genes, such as HIF-1α, c-met, u-PA, PDGF-A, and IL-8. However, IFN-α had no direct effect on migration and invasion of HCC cells., Conclusions: IFN-α has janus face of consistently suppressing HCC growth, however, promoting tumor metastasis capacity, which is of clinical indication for the scientific administration of IFN-α and the similar antiangiogenesis drugs for their dual effect on tumor growth and metastasis.

Objective: The aim of this study is to evaluate the usefulness of high-resolution computed tomography (HRCT) densitometry in the diagnosis of otosclerosis and to investigate the relationship between CT densitometry and audiometry., Methods: HRCT findings and audiometry were compared among 34 patients (34 ears, the otosclerosis group) with surgically confirmed otosclerosis between January 2007 and December 2007 and 33 patients (33 opposite normal ears, the control group) with facial paralysis diagnosed at the same period of time. Seven regions of interest (ROI) were set manually around the otic capsule on the axial slice of 0.75-mm-thick CT image. The mean CT values of these seven regions were measured. In each ROI, the mean CT value of the otosclerosis group and that of the control group were compared. Based on the CT findings, the ears with otosclerosis were classified into two groups: Group A showed no pathological CT findings; Group B showed low density around the cochlea. In the otosclerosis group, the relationship between the findings of CT and the results of audiometry was analyzed., Results: The mean CT values in the area posterior to the oval window and anterior to the oval window were significantly lower for the otosclerosis group compared with the control group (the former t=-2.030, p=0.046; the latter Z=-4.979, p<0.01). Group A consisted of 30 patients, 7 of which (23.33%) exhibited conductive hearing loss, and 23 of which (76.67%) exhibited mixed hearing loss; Group B had 4 patients, all with mixed hearing loss. For the otosclerosis group, the mean CT value in the area posterior to the oval window was positively correlated with the mean air conduction threshold (r=0.4273, p=0.0117) and with the mean air-bone gap (r=0.3995, p=0.0192)., Conclusion: Quantitative evaluation of CT with slices less than 1mm in thickness may provide important information for the diagnosis and assessment of otosclerosis which are unattainable through other methods., (Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.)

Objective: To investigate the possible mechanisms by which Somatostatin (SST) enhances the anti-tumor effect of doxorubicin (DOX) on gallbladder cancer cells., Methods: GBC-SD cells were grouped into 4 groups: SST-treated group, DOX-treated group, SST+DOX co-treated group and control group. The concentrations of SST and DOX were 75 µg/ml and 5 µg/ml based on our previous studies. In control group, cells were cultivated with phosphate buffered saline (PBS). In experimental groups, cells were cultivated with medium and the corresponding drugs. After drug treatment, cell viability was examined by MTT assay at 6, 12, 24 and 36 h respectively. Meanwhile, intracellular concentrations of doxorubicin in each group was determined by microspectrofluorimetry; Real-time polymerase chain reaction (RT-PCR) was used to determine the expressions of MDR1 mRNA in the cells at different time points and the expressions of P-gp protein, a product of MDR1 mRNA, were determined by Western blot analysis., Results: SST did not exhibit significant inhibitory effect on the proliferation of GBC-SD cells as compared to that of control group (P>0.05). SST+DOX co-treatment group and DOX showed significantly inhibitory effect on the growth of GBC-SD cells at Hour 12 post-treatment. However no statistical difference was found between SST+DOX and DOX groups. Interestingly, at Hour 24 post-treatment, SST+DOX group showed more robust inhibitory effect on GBC-SD cells as compared to DOX alone group. Moreover, SST could significantly down-regulate the expressions of MDR1 mRNA and P-gp protein. SST could increase intracellular DOX concentration. And the difference of intracellular DOX concentration between SST+DOX group and DOX group at Hour 24 was statistically significant., Conclusions: In our experiment, SST decreases the expression of MDR1 mRNA and P-gp protein so as to reduce the efflux of DOX and elevate DOX concentrations in GBC-SD cells. This eventually leads to enhanced cytotoxic effects of DOX on GBC-SD cells.

Purpose: To investigate the role of macrophages in tumor progression under sorafenib treatment and to explore whether combination of drugs that deplete macrophages improved the antitumor effect of sorafenib., Experimental Design: Tumor growth, lung metastasis, and tumor angiogenesis were observed in HCCLM3-R and SMMC7721, two human hepatocellular carcinoma xenograft nude mouse models, when treated with sorafenib (30 mg/kg daily, n = 6 per group) or a vehicle as control. Macrophage infiltration was measured in the peripheral blood and in sorafenib-treated tumor by immunohistochemistry and flow cytometry with F4/80 antibody and CD11b antibody. The effect of macrophage depletion on tumor angiogenesis and metastasis after sorafenib treatment, using two drug target macrophages, zoledronic acid (ZA) and clodrolip, was measured in the two models of hepatocellular carcinoma., Results: Although sorafenib significantly inhibited tumor growth and lung metastasis, it induced a significant increase in peripheral recruitment and intratumoral infiltration of F4/80- and CD11b-positive cells, which was accompanied with elevation of colony-stimulating factor-1, stromal-derived factor 1alpha, and vascular endothelial growth factor in the tumor and elevation of plasma colony-stimulating factor-1 and mouse vascular endothelial growth factor in peripheral blood, suggesting the role of macrophages in tumor progression under sorafenib treatment. Depletion of macrophages by clodrolip or ZA in combination with sorafenib significantly inhibited tumor progression, tumor angiogenesis, and lung metastasis compared with mice treated with sorafenib alone. ZA was more effective than clodrolip., Conclusions: Macrophages may have an important role in tumor progression under sorafenib treatment. ZA is promising when combined with sorafenib to enhance its antitumor effect.

Background: Macrophage colony-stimulating factor 1 receptor (CSF-1R) expression in hepatocellular carcinoma (HCC) and its prognostic values are unclear. This study evaluated the prognostic values of the intratumoral and peritumoral expression of CSF-1R in HCC patients after curative resection., Methods: Tissue microarrays containing material from cohort 1 (105 patients) and cohort 2 (32 patients) were constructed. Immunohistochemistry was performed and prognostic values of these and other clinicopathological data were evaluated. The CSF-1R mRNA level was assessed by quantitative real-time polymerase chain reaction in cohort 3 (52 patients)., Results: Both the CSF-1R density and its mRNA level were significantly higher in peritumoral liver tissue than in the corresponding tumor tissue. CSF-1R was distributed in a gradient in the long-distance peritumoral tissue microarray, with its density decreasing as the distance from the tumor margin increased. High peritumoral CSF-1R was significantly associated with more intrahepatic metastases and poorer survival. Peritumoral CSF-1R was an independent prognostic factor for both overall survival and time to recurrence and affected the incidence of early recurrence. However, intratumoral CSF-1R did not correlate with any clinicopathological feature. Peritumoral CSF-1R was also associated with both overall survival and time to recurrence in a subgroup with small HCCs (< or =5 cm)., Conclusions: Peritumoral CSF-1R is associated with intrahepatic metastasis, tumor recurrence, and patient survival after hepatectomy, highlighting the critical role of the peritumoral liver milieu in HCC progression. CSF-1R may become a potential therapeutic target for postoperative adjuvant treatment.

Purpose: Increasing evidence indicates that tumor-derived endothelial cells (TEC) possess a distinct and unique phenotype compared with endothelial cells (NEC) from adjacent normal tissue and may be able to acquire resistance to drugs. The aim of this study was to investigate the angiogenesis activity and response to drug treatment of TECs and NECs derived from human hepatocellular carcinoma (HCC)., Experimental Design: TECs or NECs were isolated from HCC or adjacent normal liver tissue using anti-CD105 antibody coupled to magnetic beads. The phenotypic and functional properties of endothelial cells were characterized by testing the expression of CD105, CD31, CD144, vascular endothelial growth factor receptor-1, vascular endothelial growth factor receptor-2, and von Willebrand factor, and the ability of DiI-Ac-LDL-uptake and tube formations. CD105(+) TECs were compared with CD105(+) NECs and human umbilical vein endothelial cells (HUVEC) by examining their ability to proliferate, motility, ability to adhere to tumor cells, response to tumor conditioned medium, and reactions to the chemotherapy drugs Adriamycin and 5-fluorouracil and the antiangiogenic drug sorafenib., Results: Compared with CD105(+) NECs and HUVECs, CD105(+) TECs showed increased apoptosis resistance and motility and proangiogenic properties. Meanwhile, CD105(+) TECs had a greater ability to adhere to tumor cells and survive in the tumor environment. Moreover, CD105(+) TECs acquired more resistance to Adriamycin, 5-fluorouracil, and sorafenib than CD105(+) NECs and HUVECs., Conclusions: TECs possessed enhanced angiogenic activity and resistance to chemotherapeutic drugs and an angiogenesis inhibitor, and may provide a better tool for studying tumor angiogenesis and antiangiogenesis drugs in HCC.

Background: The chemokine and bone marrow-homing receptor CXCR4 is implicated in metastases of various cancers. This study was conducted to analyze the association of CXCR4 expression with hepatocellular carcinoma (HCC) bone metastasis and patient survival., Methods: Tumor tissue from HCC patients with (n = 43) and without (n = 138) bone metastasis was subjected to immunohistochemical staining for CXCR4 using tissue microarrays. Immunoreactivity was evaluated semi-quantitatively. A receiver-operating characteristic-based approach and logistical regression analysis were used to determine the predictive value of clinicopathologic factors, including CXCR4 expression, in bone metastasis. Patient survival was analyzed by Kaplan-Meier curves and log-rank tests., Results: CXCR4 overexpression was detected in 34 of 43 (79.1%) patients with bone metastases and in 57 of 138 (41.3%) without bone metastases. CXCR4 expression correlated with (correlation coefficient: 0.551, P < 0.001) and was predictive of HCC bone metastases (AUC: 0.689; 95%CI: 0.601 - 0.776; P < 0.001). CXCR4 staining intensity correlated with the bone metastasis-free survival (correlation coefficient: -0.359; P = 0.018). CXCR4 overexpression in primary tumors (n = 91) decreased overall median survival (18.0 months vs. 36.0 months, P <0.001). Multivariable analysis identified CXCR4 as a strong, independent risk factor for reduced disease-free survival (relative risk [RR]: 5.440; P = 0.023) and overall survival (RR: 7.082; P = 0.001)., Conclusion: CXCR4 expression in primary HCCs may be an independent risk factor for bone metastasis and may be associated with poor clinical outcome.

Purpose: To examine expression profile and prognostic significance of vascular endothelial growth factor (VEGF) and its receptors in hepatocellular carcinoma (HCC) and peritumoral tissue., Methods: Expression of VEGF-A, VEGF-C, and VEGF receptor 1(VEGFR-1), VEGFR-2, and VEGFR-3 in tumor and peritumoral liver tissue was studied by immunohistochemistry in a tissue microarray from 107 patients with HCC. Unsupervised hierarchical cluster analyses were conducted to identify relevant clusters., Results: Staining of VEGF-A, VEGF-C, VEGFR-2, and VEGFR-3 was mostly found on the tumor cells and peritumoral hepatocytes, but VEGFR-1 was mostly expressed in stromal cells. In most of the cases, the expression of VEGF-A, VEGFR-1, VEGFR-2, and VEGFR-3 in was higher in peritumoral liver tissue, while VEGF-C expression was higher in tumor. Unsupervised hierarchical clustering analysis identified four prognostically different clusters, of which cluster A was classified into the "poor prognosis group," and the other three clusters were classified into the "good prognosis group" (P = 0.047). Further analysis with a set of seven markers reproduced the same four cluster groups with significantly different recurrence free probability (RFP) (P = 0.018), and the low RFP group was associated with more intrahepatic satellite lesions. Multivariate analysis showed that classification defined by seven biomarkers was of prognostic significance (P = 0.000)., Conclusions: Expression of VEGF and its receptors was higher in peritumoral tissue than in tumor in HCC. Seven biomarkers predicted patients' RFP, which consisted of tumoral expression of VEGF-A, VEGFR-1, and VEGF-C as well as peritumoral expression of VEGF-A, VEGFR-1, VEGFR-2, and VEGFR-3.

Objective: To investigate the reliability and validity of the Chinese version of voice handicap index (VHI)., Methods: The cross-cultural adaptation of health-related quality of life (HRQOL) measures was used to evaluate the Chinese version of VHI. Five hundred forty six dysphonic patients and 80 control subjects were included, 30 of patients and 20 of control subjects also had Hong Kong version VHI test simultaneously., Results: The internal consistency reliability of overall VHI scores and three subscale scores of the Chinese version of VHI were 0.8657-0.9517. The reliability coefficients (test to retest, 2-week interval) was 0.992 (P < 0.001). The correlation coefficient of overall VHI scores and three subscale scores and internal subscale scores were 0.643-0.904 (P < 0.01). There were no significant difference between the Chinese version and Hong Kong Chinese version (Z = 0.397, P = 0.691 ) with high dependability (r = 0.995, P < 0.001). The factor analysis of construct validity shows that the eigenvalue of 6 factors is above 1. The cumulative proportion was 77.24%. The loading was higher than 0.4 among every item. VHI total scores were significantly higher in dysphonic patients than in control subjects (Z = 17.69, P = 0.000). This is also true for all VHI subscores in the functional (Z = 14.14, P = 0.000), physical (Z = 17.68, P = 0.000) and emotional domains (Z = 15.50, P = 0.000)., Conclusions: The Chinese version of VHI had a good reliability and validity. It can be used to evaluate dysphonic patients.

Background: Few studies have investigated the pathologic types and prognosis of hepatocellular carcinoma (HCC) with lymph node metastasis (LNM). The purpose was to explore pathologic types and pertinent therapy of HCC with LNM., Methods: An immunohistochemical study for CK19 and OV-6 was performed on tissue microarrays of HCC with LNM (n=47) and those without LNM (n=125). The clinicopathologic factors and patient survival were analyzed., Results: Immunopositivity of CK19 and OV-6 in HCC with LNM were higher than that in 125 HCC without LNM (27.7% vs 5.6%, P=.000; 29.8% vs 12.8%, P=.009); their expressions were significantly correlated in HCC with LNM (correlation coefficient: 0.637, P=.000). The CK19 expression and tumor (T) classification of American Joint Committee on Cancer (AJCC)/International Union Against Cancer (UICC) tumor staging system were 2 independent risk factors for developing LNM (odds ratio [OR], 5.170, 95% confidence interval [CI], 1.840-14.528, P=.002; OR, 1.879, 95% CI, 1.236-2.857, P=.003). The CK19(+) group had shorter median survival (7.7 months vs 21.7 months, P=.013); CK19 expression was the independent prognostic factor for overall survival in HCC with LNM and was correlated with proliferating cell nuclear antigen labeling index and matrix metalloproteinase-9 expression (correlation coefficient: 0.484, P=.001 and 0.459, P=.001, respectively)., Conclusions: CK19 expression and AJCC/UICC T classification were 2 independent risk factors for developing LNM in HCC. CK19 expression was the independent prognostic factor for HCC with LNM. It is of clinical significance for treatment modalities to differentiate HCC with intrahepatic cholangiocarcinoma-like differentiation (CK19[+]) from one with a higher T classification (CK19[-])., (Copyright (c) 2008 American Cancer Society.)

Purpose: To investigate prognostic values of the intratumoral and peritumoral expression of macrophage colony-stimulating factors (M-CSF) in hepatocellular carcinoma (HCC) patients after curative resection., Patients and Methods: Expression of M-CSF and density of macrophages (M Phi) were assessed by immunohistochemistry in tissue microarrays containing paired tumor and peritumoral liver tissue from 105 patients who had undergone hepatectomy for histologically proven HCC. Prognostic value of these and other clinicopathologic factors was evaluated., Results: Neither intratumoral M-CSF nor M Phi density was associated with overall survival (OS) or disease-free survival (DFS). High peritumoral M-CSF and M Phi density, which correlated with large tumor size, presence of intrahepatic metastasis, and high TNM stage, were independent prognostic factors for both OS (P = .001 and P < .001, respectively) and DFS (P = .001 and P = .003, respectively) and affected incidence of early recurrence. In a small HCC subset, peritumoral M-CSF was also correlated with both OS and DFS (P = .038 and P = .001, respectively). The combination of peritumoral M-CSF and M Phi had a better power to predict the patients' death and disease recurrence (P < .001 for both)., Conclusion: High peritumoral M-CSF and M Phi were associated with HCC progression, disease recurrence, and poor survival after hepatectomy, highlighting the importance of peritumoral tissue in the recurrence and metastasis of HCC. M-CSF and M Phi may be targets of postoperative adjuvant therapy.