N6-methyladenosine (m6A) Writer WTAP Potentiates Hepatocellular Carcinoma Immune Evasion and Aerobic Glycolysis.

Hepatocellular carcinoma (HCC) is one of most prevalent malignant tumors with poor prognosis and a high mortality rate. Recent research indicates that N6-methyladenosine (m6A) and tumor immunotherapy are important factors in HCC. More research is still needed to fully understand the profound roles that m6A writer Wilms tumor 1-associated protein (WTAP) and CD8+ T cells play in the antitumor immunity that prevents HCC from progressing. According to the findings of our investigation, WTAP was significantly elevated in HCC cells and was associated with a poor prognosis. Functionally, WTAP accelerated HCC immune evasion and aerobic glycolysis while suppressing the tumor-killing ability of CD8+ T cells. On the other hand, WTAP knockdown had the opposite effect. WTAP targets the m6A site on the 3'-UTR of PD-L1 mRNA, which mechanistically increases the stability of PD-L1 mRNA. These results showed that WTAP inhibited CD8+ T cells' antitumor activity, which in turn deteriorated HCC immune evasion and aerobic glycolysis. In conclusion, our research uncovers a novel mechanism for WTAP on the tumor-killing ability of CD8+ T cells, which helps to overcome HCC immune evasion.

Application of O-RADS US combined with MV-Flow to diagnose ovarian-adnexal tumors.

PURPOSE: This study aimed to explore the application of Ovarian-Adnexal Reporting and Data System Ultrasound (O-RADS US) combined with MV-Flow (Samsung Medison Co., Ltd.) to diagnose ovarian-adnexal masses. METHODS: A total of 112 ovarian-adnexal masses (81 benign and 31 malignant) from 105 consecutive patients were analyzed. The O-RADS US and vascular index from MV-Flow (VIMV) were measured and compared with the reference standard. O-RADS US and MV-Flow were tested for consistency. RESULTS: Receiver operating characteristic curves were drawn for O-RADS US, MV-Flow, and their combination. The combined methods had the largest area under the curve (0.955), followed by O-RADS US (0.929) and MV-Flow (0.923). A mass was considered malignant when the O-RADS US classification was 5 and VIMV was ≥7.15. With this definition, MV-Flow had the highest sensitivity (87.10%), with consistent findings for the combined diagnostic methods and O-RADS US (83.87%). The specificity of the combined diagnostic methods (93.83%) was higher than that of MV-Flow (91.36%). O-RADS US had the lowest specificity (90.12%). The combined diagnostic methods had the highest coincidence rate (91.07%), and MV-Flow (90.18%) had a significantly higher coincidence rate than O-RADS US (88.39%). Both O-RADS US and MV-Flow showed good consistency among different physicians (former kappa, 0.974; latter intraclass correlation coefficient [ICC], 0.986). MV-Flow had a high consistency for the same physician (ICC, 1). CONCLUSION: O-RADS US and MV-Flow exhibited good diagnostic efficacy, and their combined diagnostic efficacy was higher than that of each individually. O-RADS US and MV-Flow can improve the diagnosis of benign and malignant ovarian-adnexal masses.

Association of radiotherapy with thoracic vertebral fractures in esophageal squamous cell carcinoma: A retrospective cohort study.

To investigate the association between radiotherapy (RT) and thoracic vertebral fractures in esophageal squamous cell carcinoma (ESCC) and explore the risk factors of thoracic vertebral fracture in ESCC who underwent RT. This retrospective cohort study including 602 consecutive ESCC patients examined the association between RT and thoracic vertebral fractures using multivariable Cox proportional hazard models and relevant risk factors of thoracic vertebral fractures based on clinical and RT parameters in patients with ESCC. Followed for a median follow-up of 24 months, 54 patients had thoracic vertebral fractures. The multivariable analysis revealed RT as an independent risk factor after adjusting for clinical risk factors. Univariable analyses associated a 5-Gy increase in vertebral dose to single vertebrae and a 1-time increase in RT fraction with higher risk of vertebral fracture. Adding RT factors (vertebral dose and fraction) and mean vertebral hounsfield unit to the Cox models containing conventional clinical risk factors significantly improved the χ2 value for predicting vertebral fractures (all P < .001). This study revealed RT, as well as increased vertebral dose and RT fractions, as a significant, consistent, and strong vertebral fracture predictor in ESCC. Combined vertebral dose, RT fractions, and vertebral hounsfield unit provided optimal risk stratification for ESCC patients.

Gonadotropic activity of a second relaxin-type peptide in starfish.

In starfish, a relaxin-like gonad-stimulating peptide (RGP) acts as a gonadotropin that triggers gamete maturation and spawning. In common with other relaxin/insulin superfamily peptides, RGP consists of an A- and a B-chain, with cross-linkages mediated by one intra- and two inter-chain disulfide bonds. In this study, a second relaxin-like peptide (RLP2) was identified in starfish species belonging to the orders Valvatida, Paxillosida, and Forcipulatida. Like RGP, RLP2 precursors comprise a signal peptide and a C-peptide in addition to the A- and B-chains. However, a unique cysteine motif [CC-(3X)-C-(10X)-C] is present in the A-chain of RLP2, which contrasts with the cysteine motif in other members of the relaxin/insulin superfamily [CC-(3X)-C-(8X)-C]. Importantly, in vitro pharmacological tests revealed that Patiria pectinifera RLP2 (Ppe-RLP2) and Asterias rubens RLP2 (Aru-RLP2) trigger shedding of mature eggs from ovaries of P. pectinifera and A. rubens, respectively. Furthermore, the potencies of Ppe-RLP2 and Aru-RLP2 as gonadotropic peptides were similar to those of Ppe-RGP and Aru-RGP, respectively, and the effect of RLP2 exhibited partial species-specificity. These findings indicate that two relaxin-type peptides regulate spawning in starfish and therefore we propose that RGP and RLP2 are renamed RGP1 and RGP2, respectively.

Upregulation of miRNA-10a-5p promotes tumor progression in cervical cancer by suppressing UBE2I signaling.

Cervical cancer (CC) is a common malignant neoplasm in gynecology. There is increasing evidence to suggest that microRNAs (miRNAs) act as crucial regulators of CC. However, whether miR-10a-5p plays a role in CC is under investigation. The aim of this stuy was to assess the miR-10a-5p expression pattern in the development of CC and investigate its downstream target. MiR-10a-5p inhibition decreased CC cell proliferation and impaired CC cell invasion and migration but enhanced apoptosis. UBE2I was a direct target of miR-10a-5p. QRT-PCR results showed a down-regulation of UBE2I in CC cells, opposing miR-10a-5p. Besides, overexpression of miR-10a-5p down-regulated UBE2I. Functional rescue experiments further indicated the miR-10a-5p-UBE2I axis was linked to CC cell growth, apoptosis and metastasis. MiR-10a-5p upregulation promotes cervical cancer development by inhibiting UBE2I. These results also predict that miR-10a-5p may be a potential target for the clinical treatment of CC.IMPACT STATEMENTWhat is already known on this subject? As a widely researched cancer-related miRNA, the overexpression of miR-10a-5p has been verified in various cancers. It has been described in a meta-analysis report that there were 42 miRNAs up-regulated and 21 miRNAs down-regulated in different stages of cervical cancer tissue versus healthy tissue.What do the results of this study add? We verified that miR-10a-5p initiates and promotes tumor cell development by decreasing UBE2I abundance. This miR-10a-5p-mediated post-transcriptional regulation of UBE2I is involved in the tumorigenesis, invasion and migration of human cervical cancer.What are the implications of these findings for clinical practice and/or further research? These findings provide mechanistic insights into how miR-10a-5p regulates cervical cancer hyper-proliferation and metastasis, as well as a new target for clinical treatment. Nevertheless, whether miR-10a-5p/UBE2I axis can be regulated by non-invasive methods need further exploration, which will be the focus of our future research.

Pectolinarigenin Suppresses LPS-Induced Inflammatory Response in Macrophages and Attenuates DSS-Induced Colitis by Modulating the NF-κB/Nrf2 Signaling Pathway.

Pectolinarigenin (PEC), a natural flavonoid present in cirsium chanroenicum and citrus fruits, has possess the distinct pharmacological activities. However, its molecular mechanisms and pharmacological effects on intestinal illness have not been elucidated. In the present study, we investigated the potential beneficial effects of pectolinarigenin (PEC) on lipopolysaccharide (LPS)-induced macrophage cells and the dextran sulfate sodium (DSS)-induced colitis model. Our findings showed that PEC pretreatment inhibits the LPS-induced nuclear factor-kappa B (NF-κB) activation by interfering with the degradation of IκB-α. Further, increased Nrf2 protein expression was reported on PEC treated RAW 264.7 and THP1 cell lines. In addition, we revealed that PEC mediated the NF-κB/Nrf2 pathway regulation, which in turn inhibits the synthesis of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), interleukin-1beta (IL-1ß), and tumor necrosis factor-alpha (TNF-α) on RAW 264.7 and THP1 cells. Furthermore, PEC dose-dependently reduced the DSS-induced inflammation in the colon by regulating NF-κB/Nrf2 signaling pathway and enhancing the myeloperoxidase (MPO) activity and redox regulators such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and lipid peroxidation byproduct malondialdehyde (MDA) in DSS-induced inflamed colon. Similarly, we reported the minimal pathological damages in the PEC-treated mice colon, as well as increase goblet cell population and mucin-2 production. In conclusion, our findings demonstrate that PEC reduces the DSS-induced colitis in mice by regulating the NF-κB/Nrf2 pathway. Thus, PEC might be a promising therapeutic agent for the treatment of inflammatory bowel disease.

Diagnostic Value of Two-Dimensional Transvaginal Ultrasound Combined with Contrast-Enhanced Ultrasound in Ovarian Cancer.

Objective: Explore the value of two-dimensional transvaginal ultrasound combined with contrast-enhanced ultrasound in the differential diagnosis of ovarian cancer, so as to provide the basis for clinical diagnosis and treatment of ovarian cancer. Methods: A total of 100 suspected ovarian cancer patients who were admitted to our hospital from January 2019 to December 2021 were selected as the research subjects, including 62 ovarian cancer patients (ovarian cancer group) and 38 ovarian benign tumor patients (benign group). Two-dimensional vaginal ultrasound and contrast-enhanced ultrasound were performed in both groups. The differences in PI, RI, EDV, PSV, and VM parameters of the two groups as well as those of patients with ovarian cancer of different grades were compared. Record the contrast-enhanced ultrasound parameters such as AT, TTP and IMAX, and determine the diagnostic value. Results: The PI and RI of the ovarian cancer group were lower than those of the benign ovarian tumor group, and the EDV, PSV and VM of the ovarian cancer group were higher than those of the benign ovarian tumor group (p < 0.05). The PI and RI of the patients in stage I-II of the ovarian cancer group were higher than those in stage III-IV, and the EDV, PSV and VM were lower than those in the patients in stage III-IV, with statistical significance (p < 0.05). The results of contrast-enhanced ultrasound showed that the AT and TTP values in the ovarian cancer group were significantly shorter than those in the benign group, and the peak intensity was significantly higher than that in the benign group, and the differences were statistically significant (p < 0.05). The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of two-dimensional ultrasound combined with contrast-enhanced ultrasound in the diagnosis of ovarian cancer were high, 95.16%(59/62), 86.84%(33/38), 92.19%(59/64), 91.67%(33/36) and 92.00%(92/100), respectively. Conclusion: Contrast-enhanced ultrasound to some extent makes up for the deficiencies of conventional ultrasound, is helpful to detect early ovarian cancer, and can be used for the differential diagnosis of small ovarian tumors with difficult two-dimensional ultrasound diagnosis. Two-dimensional ultrasound combined with contrast-enhanced ultrasound can effectively improve the detection rate and differential diagnosis value of ovarian cancer, which is of great significance in the early diagnosis and differentiation of ovarian cancer.

San Pian decoction can treat nitroglycerin-induced migraine in rats by inhibiting the PI3K/AKT and MAPK signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: San Pian decoction (SPD), a traditional Chinese medicine preparation composed of eight herbs, has been reported to alleviate migraine. However, its active ingredients and the potential mechanism of action remains unclear. The purpose of this study was to comprehensively analyze SPD for the treatment of chronic migraine based on pharmacological direction and to identify the active ingredients and pharmacological mechanism of SPD in the treatment of migraine. MATERIALS AND METHODS: The active components in SPD were identified by AB SCIEX quadrupole time-of-flight mass spectrometer, and the prediction targets and pharmacological networks related to migraine were constructed. The mechanism of SPD in treating migraine was studied through network pharmacology, which was further verified using pharmacological experiments. RESULTS: A total of 489 targets of 26 compounds were identified. Based on Venn analysis, we found 117 intersection targets between SPD and migraine, that is, these targets were related to the treatment of migraine. Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the treatment of migraine using SPD was related to the PI3K/AKT and MAPK signaling pathways. The effect of SPD on migraine was verified by measuring the levels of the inflammatory factors, nitric oxide (NO), interleukin (IL-6), endothelin (ET),5-hydroxytryptamine(5-HT), indoleamine 2,3-dioxygenas (IDO), tumor necrosis factor (TNF-α) and calcitonin gene-related peptide (CGRP). Lastly, real-time polymerase chain reaction and western blotting were used to verify gene and protein expression in the PI3K/AKT and MAPK signaling pathways. Expression of the genes P38, JNK, ERK, PI3K and AKT, and the protein expression of p-P38, p-JNK, p-ERK, p-AKT and p-PI3K were significantly downregulated. Our findings indicated that SPD could prevent inflammation by regulating the inflammatory cytokines and key genes and proteins in the PI3K/AKT and MAPK signaling pathways to treat migraine. CONCLUSION: Our findings reveal that SPD could treat nitroglycerin-induced migraine by regulating p-AKT, p-pI3k, p-p38, p-ERK, p-JNK, IL-6, and TNF-α inflammatory factors in the PI3K/AKT and MAPK signaling pathways.

Metabolomic biomarkers for hepatocellular carcinoma: A systematic review.

BACKGROUND: Hepatocellular carcinoma (HCC) is a highly malignant cancer which lack of effective diagnosis and prognosis biomarkers, therefore surging studies focused on the metabolite candidates for HCC. The current study was designed to systematically review the metabolic studies for HCC, summarize the current available evidence and provide implication for further studies within this area. By systematically screening Pubmed and Embase, and eligibility assessment, we eventually included 55 pieces of studies. After summarized their characteristics, we reviewed them by 3 parts, regarding to the different biofluid they carried out the experiments. By collecting the candidates from all the included studies, we carried out pathway enrichment to see the representative of the reported candidates, as expected the pathway consistent with the current knowledge of HCC. Next, we conduct quality assessment on the included studies. Only 36% of the current evidence grouped as high quality, indicating the quality of metabolic studies needs further improvement.

Individually designed fully covered self-expandable metal stents for pediatric refractory benign esophageal strictures.

To share our institutional experience of placing individually designed fully covered self-expandable metal stents (FCSEMSs) for the treatment of refractory benign esophageal strictures (RBESs) in pediatric patients. A 10-year retrospective study between May 2009 and July 2020 that includes 14 children with RBESs who were treated with individually designed FCSEMSs. Patients were followed-up regularly after stent placement to observe the improvement of vomiting and dysphagia, changes in stenosis diameter and complications. A total of 20 stents were successfully placed in 14 patients. During a follow-up period ranging from 5 to 83 months, except for one 4-year-old child who could not endure chest pain, the remaining 13 patients all benefited from stenting. Their Ogilvie & Atkinson scores improved from grade III-IV to grade 0-I, and the diameters of the stenosis' were enlarged from 2-5 mm to 9-14 mm. Two patients developed restenosis and granulation tissue hyperplasia was found in 2 patients and stent migration and malapposition in 2 patients with esophageal perforations that required further endoscopic intervention. The use of FCSEMS for RBES is safe and effective in selected pediatric patients. Rationally designed stents and timely management of postoperative complications are critical to ensure the success of this new method.

Bulleyaconitine A Effectively Relieves Allergic Lung Inflammation in a Murine Asthmatic Model.

BACKGROUND Bulleyaconitine A (BLA) has been widely used as analgesic against chronic inflammatory pain in China. However, its potential therapeutic role in asthma remains unclear. The purpose of this study was to investigate the effect of BLA on airway inflammation in mice with allergic asthma. MATERIAL AND METHODS Specific-pathogen-free (SPF) female Balb/c mice were randomly divided into the following 6 groups: (1) Control group (NC), (2) Asthma group (AS), (3) BLA-L group, (4) BLA-M group, (5) BLA-H group, and (6) Dexamethasone group. An asthma mouse model was established by administration of ovalbumin (OVA) and mice were sacrificed within 24 h after the last challenge. Enzyme-linked immunosorbent assay (ELISA) method was used to determine the relative expression levels of IgE and IgG in mouse serum. In addition, bronchoalveolar lavage fluid (BALF) was collected and IL-4, TNF-α, and MCP-1 levels were determined by ELISA. Furthermore, eosinophils, lymphocytes, and macrophages in BALF were classified and analyzed, and inflammatory cell infiltration in the airways of mice was determined by hematoxylin-eosin (HE) staining. The expression of NF-κB1 and PKC-δ in mouse lung tissue was determined by Western blot analysis. RESULTS The levels of serum IgE and IgG in BLA- or Dex- treated mice were significantly reduced compared to those in the asthma (AS) group (P<0.01), whereas the levels of cytokines IL-4, TNF-α, and MCP-1 were significantly decreased (P<0.01). HE-staining showed that BLA significantly reduced inflammatory cell infiltration and mucus secretion in lung tissue. Moreover, BLA inhibited the expression of NF-κB1 and PKC-d via the NF-κB signaling pathway in the lung. CONCLUSIONS Our data show that BLA activates PKC-δ/NF-κB to reduce airway inflammation in allergic asthma mice.

The diagnostic value of intracavitary electrocardiogram for verifying tip position of peripherally inserted central catheters in cancer patients: A retrospective multicenter study.

PURPOSE: To evaluate the feasibility and accuracy of intracavitary electrocardiogram for verifying tip position of peripherally inserted central catheters in cancer patients during follow-up period. METHODS: From March 2015 to October 2015, 126 patients involved in eight hospitals who underwent peripherally inserted central catheter placement received intracavitary electrocardiogram and chest X-ray to verify position of the catheter tip during follow-up period. Their intracavitary electrocardiogram was compared with surface electrocardiogram to judge catheter tip landing zone in one of three different anatomical zones. The amplitude of intracavitary electrocardiogram P wave and the ratio of intracavitary electrocardiogram P wave/surface electrocardiogram P wave were measured and showed correlation with catheter tip position confirmed by chest X-ray. Based on chest X-ray principle, all the cases were assigned into three intracavitary electrocardiogram groups to explore the optimal cut-off values for intracavitary electrocardiogram P wave and intracavitary electrocardiogram P wave/surface electrocardiogram P wave by analyzing the receiver operating characteristic. RESULTS: No technique-related complications or adverse events occurred in this study. The matching rate between intracavitary electrocardiogram and chest X-ray method was 93.7%. The optimal cut-off values for intracavitary electrocardiogram P wave were set from 3.15 to 3.75 mV, and intracavitary electrocardiogram P wave/surface electrocardiogram P wave from 1.65 to 3.25. CONCLUSIONS: It is demonstrated in this retrospective multicenter study that the intracavitary electrocardiogram method for verifying tip position of peripherally inserted central catheter during follow-up period is feasible and accurate in all adult patients with cancer.

Superior success rate of intracavitary electrocardiogram guidance for peripherally inserted central catheter placement in patients with cancer: A randomized open-label controlled multicenter study.

BACKGROUND: Intracavitary electrocardiogram (IC ECG) guidance emerges as a new technique for peripherally inserted central catheters (PICCs) placement and demonstrates many potential advantages in recent observational studies. AIMS: To determine whether IC ECG-guided PICCs provide more accurate positioning of catheter tips compared to conventional anatomical landmarks in patients with cancer undergoing chemotherapy. METHODS: In this multicenter, open-label, randomized controlled study (ClinicalTrials.gov number, NCT02409589), a total of 1,007 adult patients were assigned to receive either IC ECG guidance (n = 500) or anatomical landmark guidance (n = 507) for PICC positioning. The confirmative catheter tip positioning x-ray data were centrally interpreted by independent radiologists. All reported analyses in the overall population were performed on an intention-to-treat basis. Analyses of pre-specified subgroups and a selected large subpopulation were conducted to explore consistency and accuracy. RESULTS: In the IC ECG-guided group, the first-attempt success rate was 89.2% (95% confidence interval [CI], 86.5% to 91.9%), which was significantly higher than 77.4% (95% CI, 73.7% to 81.0%) in the anatomical landmark group (P < 0.0001). This trend of superiority of IC ECG guidance was consistently noted in almost all prespecified patient subgroups and two selected large subpopulations, even when using optimal target rates for measurement. In contrast, the superiority nearly disappeared when PICCs were used via the left instead of right arms (interaction P-value = 0.021). No catheter-related adverse events were reported during the PICC intra-procedures in either group. CONCLUSIONS: Our findings indicated that the IC ECG-guided method had a more favorable positioning accuracy versus traditional anatomical landmarks for PICC placement in adult patients with cancer undergoing chemotherapy. Furthermore, there were no significant safety concerns reported for catheterization using the two techniques.

Metabolism and bioactivation of the tricyclic antidepressant amitriptyline in human liver microsomes and human urine.

AIM: Amitriptyline is a widely used tricyclic antidepressant, but the metabolic studies were conducted almost 20 years ago using high-performance liquid chromatography coupled with ultraviolet detector or radiolabeled methods. RESULTS: First, multiple ion monitoring (MIM)- enhanced product ion (EPI) scan was used to obtain the diagnostic ions or neutral losses in human liver microsome incubations with amitriptyline. Subsequently, predicted multiple reaction monitoring (MRM)-EPI scan was used to identify the metabolites in human urine with the diagnostic ions or neutral losses. Finally, product ion filtering and neutral loss filtering were used as the data mining tools to screen metabolites. Consequently, a total of 28 metabolites were identified in human urine after an oral administration using LC-MS/MS. CONCLUSION: An integrated workflow using LC-MS/MS was developed to comprehensively profile the metabolites of amitriptyline in human urine, in which five N-acetyl-l-cysteine conjugates were characterized as tentative biomarkers for idiosyncratic toxicity.

Combination of 2- and 3-dimensional contrast-enhanced transvaginal sonography for diagnosis of small adnexal masses.

OBJECTIVES: The purpose of this study was to evaluate the efficacy of the combination of 2-dimensional (2D) and 3-dimensional (3D) contrast-enhanced sonography in discriminating between benign and malignant small adnexal masses. METHODS: Selected patients were evaluated with both 2D and 3D contrast-enhanced sonography after conventional sonography before undergoing any surgery. Time-intensity curves for 2D contrast-enhanced sonography were constructed by using contrast-enhanced sonographic software. A vascular perfusion characteristic analysis was achieved by 2D and 3D contrast-enhanced sonography. Results were finally verified by surgery. RESULTS: Forty-seven cases of benign and 10 cases of malignant small adnexal masses were discovered. Significant differences in perfusion patterns, time-intensity curve shapes for 2D contrast-enhanced sonography, grayscale contrast-enhanced sonography, and blood flow imaging on 3D contrast-enhanced sonography were observed between benign and malignant masses (P< .05). Two-dimensional contrast-enhanced sonography, 3D contrast-enhanced sonography, parallel combination of 2D and 3D contrast-enhanced sonography, and serial combination of 2D and 3D contrast-enhanced sonography all reached diagnostic sensitivity of 100% for discriminating benign from malignant masses, whereas specificity values were 61.7%, 63.8%, 68.1%, and 57.4%, respectively. Areas under the receiver operating characteristic curves were 0.809, 0.819, 0.840, and 0.787. CONCLUSIONS: Two-dimensional contrast-enhanced sonography is of high value in distinguishing malignant from benign small adnexal masses; 3D contrast-enhanced sonography provides richer and more useful information for evaluation of these masses. Diagnostic sensitivity of 100% can be achieved when using a serial combination of 2D and 3D contrast-enhanced sonography, although specificity needs further improvement.

Hepatitis B virus infection in clustering of infection in families with unfavorable prognoses in northwest China.

Hepatitis B virus (HBV) infection and its associated liver diseases have characteristics of familial clustering in China. However, the reasons for this are not understood fully. To address this issue, the prevalence HBV infection and the characteristics of unfavorable prognoses in clustering of infection in families in northwest China were investigated. Families with clustering of infection and unfavorable prognoses were enrolled, and general information and serum samples were collected. The clinical features and sequelae of HBV infection were compared among the blood relatives (including the first-, second-, and third-degree blood relatives) and spouses using the chi-square test or Fisher's exact test. A total of 102 clusterings of infection families with unfavorable prognoses were interviewed. In the first-, second-, and third-degree blood relatives and spouses, the prevalences of cirrhosis of the liver were 29.2%, 11.9%, and 8.7%, respectively, while those of hepatocellular carcinoma (HCC) were 21.8%, 1.4%, and 4.3%, respectively (P<0.05). The mean ages of the onset of cirrhosis of the liver in the first-, second-, and third-degree blood relatives and spouses were 57 ± 9.91, 47 ± 9.96, 38 ± 10.35, and 57 ± 8.49 years, respectively, while the mean ages of the onset of HCC were 60 ± 7.92, 49 ± 8.57, 41 ± 3.54, and 50 ± 0 years, respectively, (P<0.05). The first-, second-, and third-degree blood relatives from clustering of infection in families with unfavorable prognoses had prevalences of cirrhosis or HCC in descending order of relationship. The findings suggest that genetic factors may be associated with a familial tendency for cirrhosis of the liver and HCC.

Value of three-dimensional contrast-enhanced ultrasound in the diagnosis of small adnexal masses.

The main purpose of this study was to determine whether three-dimensional contrast-enhanced ultrasound (3D-CEUS) can provide useful information to distinguish malignant from benign adnexal masses (≤4 cm). Forty-seven patients with 51 adnexal masses were examined with 3D-CEUS. The sonographic features of masses were analyzed. All diagnoses were confirmed by surgical pathology and long-term follow-up results. The 51 masses included 43 benign and 8 malignant lesions. On 3D-CEUS images, benign lesions appeared as round structures formed by sparse and straight capillary vessels. Malignant lesions showed irregular stereo structures with dense and tortuous vascular distribution. A 3D-CEUS scoring system was established. There were no statistically significant differences in scores at each time point between the 20th and 70th seconds, and the area under the receiver operating characteristic curve for this time period was the largest (0.995). A cut-off score of 8 was established, with scores ≥8 being suggestive of malignancy. The 3D-CEUS scoring system had a high sensitivity (100%) and specificity (98%). 3D-CEUS is likely to be the new tool to distinguish malignant from benign small adnexal masses and diagnose early ovarian cancer.

Involvement of ERK, Bcl-2 family and caspase 3 in recombinant human activin A-induced apoptosis in A549.

BACKGROUND: Activins are members of the transforming growth factor-beta (TGF-beta) superfamily. Previous studies have shown that activin A may have a central role in regulating both apoptosis and proliferation. However, direct studies of recombination human activin A on human NSCLC A549 cells have not yet been reported. The purpose of this study was to investigate whether activin A could induce apoptosis in A549 cells and the possible mechanisms via which it worked. METHODS: Cellular apoptosis induced by activin A was detected by TUNEL assay and the levels of protein expression were detected by western blot. RESULTS: Recombination human activin A induced apoptosis in human NSCLC A549 cells in a concentrate-dependent manner. Activin A-induced A549 apoptosis was accompanied by the up-regulation of Bax, Bad and Bcl-Xs and down-regulation of Bcl-2. Moreover, activin A treatment increased the expression of its typeII receptors, activated ERK and caspase 3 in A549. These results clearly demonstrate that the induction of apoptosis by activin-A involves multiple cellular/molecular pathways and strongly suggest that pro- and anti-apoptotic Bcl-2 family proteins and caspase 3 participate in activin A-induced apoptotic process in A549 cells. On the other hand, activin A treatment had little effect on primary human small airway epithelial cells (SAECs). CONCLUSION: Recombination human activin A induced apoptosis in A549 cells, at least partially, through ERK and mitochondrial pathway. The result that activin A did not affect the normal SAEC revealed activin A might be considered as a potential anticancer agent and worthy of further studies.

Budesonide/formoterol decreases expression of vascular endothelial growth factor (VEGF) and VEGF receptor 1 within airway remodelling in asthma.

INTRODUCTION: Angiogenesis and microvascular remodelling may play a vital role in the chronic inflammatory process within asthma. One of the most important factors involved in angiogenesis is vascular endothelial growth factor (VEGF). In this study we hypothesised that an increased expression of VEGF may be involved in airway remodelling in asthma patients. To this end, we compared the histology and expression levels of VEGF and one of its receptors (VEGFR1) in bronchial tissues of patients with asthma compared with control patients. We also investigated the effect of treatment with budesonide/formoterol (Symbicort; AstraZeneca, Lund, Sweden) in the relationship between VEGF and airway remodelling. METHODS: Bronchial tissues were obtained from patients attending the West China Hospital from April to November 2006. Thirteen patients were diagnosed with moderate asthma and 10 others were treated as control. Histological and immunohistochemical comparisons between asthmatic and control patients were made at baseline, and (for asthmatic subjects) following 6 months of treatment with budesonide/formoterol. RESULTS: Compared with control patients, asthmatic patients had significantly decreased respiratory parameters, including forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV(1)) (% predictive). Furthermore, patients with asthma had submucosal gland hyperplasia, increased smooth muscle mass, increased subepithelial fibrosis and neovascularisation. Asthmatic patients also exhibited increased expression of VEGF and VEGFR1 within epithelial cells. The increased expression of VEGF and its receptor correlated well with airway remodelling, airflow obstruction and airway hyper-responsiveness. After treatment with budesonide/formoterol for 6 months, the expression of VEGF and VEGFR1 was decreased, with correlatory decreased airway remodelling in patients with asthma. CONCLUSION: The increased expression of VEGF and VEGFR(1) in asthmatic patients is accompanied by an increased number and size of blood vessels in asthmatic airways, as well as airway remodelling. Budesonide/formoterol therapy for 6 months can decrease the expression of VEGF and VEGFR(1) alongside airway remodelling in asthma. The inhibition of VEGF and its receptor may be a good therapeutic strategy for asthma.

Macrolides attenuate mucus hypersecretion in rat airways through inactivation of NF-kappaB.

BACKGROUND AND OBJECTIVE: To examine the effect of a 14-membered ring macrolide on airway mucus hypersecretion in rats treated with LPS. METHODS: Mucus hypersecretion in rat airways was induced by intratracheal instillation of LPS. Rats treated with or without LPS were administered roxithromycin (1-10 mg/kg), josamycin (10 mg/kg) or amoxicillin (40 mg/kg), orally for 4 days. Expression of Muc5ac, nuclear factor (NF)-kappaB, and the mitogen-activated protein (MAP) kinases p38 and ERK1/2 in bronchial epithelium were detected by RT-PCR, immunohistochemistry or western blotting. Mucins, IL-1beta, IL-8 and tumour necrosis factor (TNF)-alpha in BAL fluid were assayed by enzyme-linked lectin assay and ELISA. RESULTS: LPS significantly induced the expression of Muc5ac mRNA and protein in bronchial epithelium, increased the release of mucins, IL-1beta, IL-8 and TNF-alpha, and increased neutrophil numbers in BAL. Moreover, LPS increased staining for NF-kappaB in the cytoplasm as well as nuclear translocation of NF-kappaB in airway epithelial cells. Upregulated expression of Muc5ac mRNA correlated positively with NF-kappaB activation and the levels of cytokines (P < 0.05). Roxithromycin (5 and 10 mg/kg) significantly attenuated bronchial Muc5ac expression and NF-kappaB nuclear translocation stimulated by LPS, and reduced neutrophil numbers, mucins and inflammatory cytokines in BAL (P < 0.05). However, LPS-stimulated expression of p38 and ERK1/2 in airway epithelium was not affected by roxithromycin. Josamycin and amoxicillin had no effects on Muc5ac expression, NF-kappaB activation or cytokine release. CONCLUSIONS: Roxithromycin inhibits the pulmonary inflammatory response and airway mucus hypersecretion induced by LPS. The inhibitory effect of roxithromycin on airway mucus hypersecretion may be mediated through reduction of NF-kappaB activation, neutrophil infiltration and release of inflammatory cytokines in the lung.