DNA damages in hepatocytes are amended by an inflammation-driven rescue repair mechanism in chronic hepatitis B.

BACKGROUND: Our previous study has shown that intrahepatic necroinflammation favors the eliminations of HBV integration and clonal hepatocytes. Here, the effect of inflammation on host DNA damage eliminations in liver biopsy tissues from patients with chronic hepatitis B (CHB) was further investigated. METHODS: DNA damage markers, histone γ-H2AX and phosphorylated heterochromatin protein 1γ (p-HP1γ), and senescent marker p21 were detected using immunohistochemical and immunofluorescent assays in liver biopsy samples from 69 CHB patients and 12 liver cirrhosis (LC) patients. Twenty paired hepatocellular carcinoma (HCC) surgical samples were used as controls. RESULTS: Both γ-H2AX and p-HP1γ were sensitively detected in nuclear and cytoplasmic/nuclear patterns. Nuclear γ-H2AX was superior as a DNA damage marker in hepatocytes. The level of nuclear γ-H2AX in CHB, comparable to those in LC and HCC, was correlated with liver fibrosis and coexisted with the senescent marker p21. However, hepatocytes carried an alleviated level of DNA damages, which was associated with the level of cytoplasmic γ-H2AX. Cytoplasmic γ-H2AX chiefly occurred in hepatocytes near necroinflammatory foci, was correlated with liver inflammation and usually indicated the decrease or disappearance of nuclear γ-H2AX. The lack of cytoplasmic γ-H2AX together with the high level of nuclear γ-H2AX was associated with the progression from large cell changes/dysplasia to small cell changes/dysplasia. CONCLUSIONS: Hepatocytes in CHB already carry massive DNA damages and undergo cellular senescence. The DNA damages in those senescent hepatocytes are histopathologically demonstrated to be amended by a novel cytoplasmic γ-H2AX-indicated and inflammation-driven rescue repair mechanism, which may be involved in hepatocarcinogenesis if it works improperly.

Hypoxaemia risk in pediatric flexible bronchoscopy for foreign body removal: a retrospective study.

BACKGROUND: Hypoxemia represents the most prevalent adverse event during flexible bronchoscopy procedures aimed at foreign body retrieval in pediatric patients; if not expeditiously managed, it carries the potential for cardiac or respiratory arrest. The specific risk factors contributing to the occurrence of hypoxemia during foreign body FB removal via bronchoscopy have yet to be definitively established. METHODS: This retrospective study included a cohort of 266 pediatric subjects from January 1, 2015, to December 31, 2022, who underwent flexible bronchoscopy for the purpose of FB extraction. In this cohort, the supraglottic airway was used to connect the anesthesia apparatus during the removal procedure. RESULTS: In total, 45 of the pediatric patients (16.9%) experienced episodes of hypoxemia during the FB removal procedure. Multivariate analysis revealed that the following factors were significantly associated with the occurrence of hypoxemia: an operation time exceeding 60 min (odds ratio [OR] 8.55; 95% confidence interval [CI] 3.82-19.13), a maximum diameter exceeding 7 mm (OR 5.03; 95% CI, 2.24-11.29), and the presence of radiological evidence indicating pneumonia (OR 2.69; 95% CI, 1.27-5.69). CONCLUSION: During flexible bronchoscopy procedures aimed at FB removal in pediatric patients, there is an increased susceptibility to hypoxemia. Factors including extended operation duration, larger FB dimensions, and radiographic evidence suggestive of pneumonia significantly contribute to a heightened risk of hypoxemia.

Assessment of the Risk of Malnutrition or Frailty Among Patients Undergoing Liver Transplantation: A Hospital-Based Prospective Study.

Objective: We aimed to explore the status of nutritional and frailty in patients undergoing liver transplantation and the associated influencing factors. Methods: We conducted a follow-up analysis of 44 patients who underwent liver transplantation between 2021 and 2022. We followed up and recorded the nutritional status and risk of weakness at different time-points (days 1, 2, 3, 6, 9, and 12) postoperatively. Patient information regarding demographics, physical examination, medical history, and perioperative blood tests were collected. Binary logistic regression was applied to identify risk factors for weakness after liver transplantation. Results: The cohort comprised 44 liver transplant recipients, with a mean age of 47.66 years (standard deviation=9.49 years). Initial analysis revealed that, compared to the group without nutritional risks, the group with nutritional risks displayed elevated age and preoperative blood ammonia levels one week post-surgery. Moreover, this group had reduced levels of albumin and total bile acid preoperatively. Patients with preoperative nutritional risks were also prone to similar risks 2 weeks postoperatively. Further, a correlation was observed between preoperative pulmonary infections and increased frailty risk 6 days postoperatively. At both 9 and 12 days postoperatively, patients with frailty risk exhibited higher preoperative white blood cell counts and ammonia levels than those without. Multivariable analysis, controlling for confounding factors, indicated a significant association between preoperative nutritional status and nutritional risk 2 weeks postoperatively, as well as a link between preoperative white blood cell count and frailty risk at 12 days postoperatively. Conclusion: There was a significant correlation between preoperative nutritional status and nutritional risk 2 weeks after liver transplantation, and preoperative white blood cell count was an independent risk factor for weakness 12 days postoperatively. Preoperative nutritional management for patients could potentially mitigate the likelihood of adverse clinical outcomes.

Novel Bipolar High-Voltage Pulsed Radiofrequency Targeting the Cervical Sympathetic Chain for Treating Acute Herpetic Neuralgia.

INTRODUCTION: Postherpetic neuralgia (PHN) in the oral, maxillofacial, neck, and upper limb regions is a refractory neuropathic pain and severely affects the quality of life of patients. Because of the absence of ideal treatments for this condition, relieving pain in the acute stage and preventing the occurrence of PHN are of great clinical significance. However, the optimal intervention for this acute herpetic neuralgia remains obscure. OBJECTIVES: This study aimed to investigate whether bipolar high-voltage pulsed radiofrequency (PRF) targeting the cervical sympathetic chain could effectively treat acute herpetic neuralgia in the oral, maxillofacial, neck, and upper limb regions and reduce the incidence of PHN. MATERIALS AND METHODS: A total of 60 patients with acute herpetic neuralgia in the oral, maxillofacial, neck, and upper limb regions were enrolled. The radiofrequency group (n = 30) received bipolar high-voltage PRF under ultrasound guidance at the level of the transverse processes of C6 and C7 to modulate the cervical sympathetic chain. In the sham group (n = 30), the electrodes were simply placed at the same position as in the radiofrequency group, but no radiofrequency energy was applied. The same treatment was repeated in each group after 72 hours. If patients were reported to have a visual analog scale (VAS) score ≥4, they would receive oral tramadol and gabapentin as rescue analgesics. The VAS score, Pittsburgh Sleep Quality Index (PSQI), 36-Item Short Form Health Survey (SF-36) score, use of tramadol and gabapentin, incidence of PHN, and adverse reactions were recorded to assess the effect and safety of therapy during three months of follow-up. RESULTS: Decreased VAS scores, PSQI scores, and improved SF-36 scores were detected in the two groups at different time points after treatment (all p < 0.05). The VAS scores, PSQI scores, use of tramadol and gabapentin, and incidence of PHN were significantly lower, whereas the SF-36 scores were significantly higher in the radiofrequency group than in the Sham group (all p < 0.05). No serious adverseness related to the treatment was detected in either group. CONCLUSIONS: Bipolar high-voltage PRF treatment targeting the cervical sympathetic chain could effectively relieve acute herpetic neuralgia in the oral, maxillofacial, neck, and upper limb regions and reduce PHN incidence. The efficacy and safety of this novel treatment make it worthy of recommendation for clinical application.

Development and clinical application of a rapid and visual loop-mediated isothermal amplification test for tetM gene in Clostridioides difficile strains cultured from feces.

OBJECTIVES: In this study, we aimed to develop a rapid and visual loop-mediated isothermal amplification (LAMP) assay targeting the tetM gene in Clostridioides difficile strains cultured from feces. METHODS: Primers were designed to recognize the tetM gene in C. difficile by LAMP, using turbidity and visual detection. The sensitivity and specificity of LAMP primers were determined. In addition, we conducted both LAMP and polymerase chain reaction (PCR) for the tcdA, tcdB, cdtA, cdtB, ermB, and tetM genes in 300 toxigenic C. difficile strains cultured from feces. RESULTS: The target DNA was amplified and visualized within 60 minutes at a temperature of 62°C. A total of 26 bacterial strains were found negative for tetM, which manifested high specificity of the primers. The detection limit of LAMP was 36.1 pg/µl, which was 100-fold more sensitive than PCR. The positive rate of tetM in toxigenic C. difficile strains cultured from feces was 93.3% by both LAMP and PCR. The proportion of toxin types in those C. difficile strains was 95.7% for A+B+CDT-, 4% for A-B+CDT-, and 0.3% for A+B+CDT+, respectively. CONCLUSION: This is the first study examining the tetM gene by LAMP in C. difficile strains cultured from feces. Its high specificity, sensitivity, and visual detection make the new assay a powerful diagnostic tool for rapid testing.

Risk factors associated with reintubations in children undergoing foreign body removal using flexible bronchoscopy: a single-center retrospective cross-sectional study.

BACKGROUND: Reintubation is a severe complication during foreign body (FB) removal that uses flexible bronchoscopy. OBJECTIVE: To investigate the incidence and risk factors for reintubations in children undergoing FB extraction by flexible bronchoscopy in a single center. DESIGN: A retrospective cross-sectional study. SETTING: All children with foreign body aspiration at Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University from January 2015 to December 2020. PATIENTS: Children with FB removal using a flexible bronchoscopy were enrolled in the trial according to the inclusion criteria. MEASUREMENTS: Both multivariable and logistic regression analyses were used to analyze the association between characteristic data and reintubations. The results were presented as odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: In total, 244 patients met with the inclusion criteria and were included in the analysis. Among those participants, 28 children (11.5%) underwent reintubations after FB removal by flexible bronchoscopy. Independent factors associated with reintubations were identified as operative time ≥ 60 min [OR: 3.68, 95% CI (1.64-8.82)] and ASA ≥ III [OR: 5.7, 95% CI (1.23-26.4)]. CONCLUSIONS: Children undergoing FB removal by a flexible bronchoscopy may encounter with a high incidence of postoperative reintubations. Both long operative duration and a severe physical status cause a growing risk of reintubations.

Quinoa Secondary Metabolites and Their Biological Activities or Functions.

Quinoa (Chenopodium quinoa Willd.) was known as the "golden grain" by the native Andean people in South America, and has been a source of valuable food over thousands of years. It can produce a variety of secondary metabolites with broad spectra of bioactivities. At least 193 secondary metabolites from quinoa have been identified in the past 40 years. They mainly include phenolic acids, flavonoids, terpenoids, steroids, and nitrogen-containing compounds. These metabolites exhibit many physiological functions, such as insecticidal, molluscicidal and antimicrobial activities, as well as various kinds of biological activities such as antioxidant, cytotoxic, anti-diabetic and anti-inflammatory properties. This review focuses on our knowledge of the structures, biological activities and functions of quinoa secondary metabolites. Biosynthesis, development and utilization of the secondary metabolites especially from quinoa bran were prospected.

Risk of upgrading from prostate biopsy to radical prostatectomy pathology: Is magnetic resonance imaging-guided biopsy more accurate?

Background: This study compared magnetic resonance imaging-guided biopsy (MRI-GB) and transrectal ultrasound guided biopsy (TRUS-GB) with the final histology of the radical prostatectomy (RP) specimen. Methods: Our subjects were 229 patients with prostate cancer (PCa), proven histopathologically using MRI-GB or TRUS-GB, who underwent RP at our center between December 2015 and December 2016. The main group included 92 patients who underwent MRI-GB and the control group included 137 patients who underwent 12-core TRUS-GB. Histological findings for RP specimens were compared with those from biopsies. We also evaluated predictors of upgraded Gleason score (GS), using uni- and multivariate analyses. Results: Upgraded GS between biopsy and RP specimen occurred to 22.7% (52/229) of the cohort overall. In univariate analysis, prostate-specific antigen density (PSAD) (P<0.001), prostate volume (PV) < 30 ml (P<0.001), biopsy modality (P=0.027), biopsy GS (P=0.032) and measured MRI lymph node metastasis (P=0.018) were prognostic factors. Multivariate logistic regression analysis showed PV < 30 ml (P<0.001) and biopsy modality (P=0.001) were independent predictors of upgraded GS. Conclusions: MRI-GB may enhance the diagnostic accuracy of prostate cancer detection in final histopathology with lower rate of upgraded GS than TRUS-GB. Also, PV < 30 ml and biopsy modality were independent predictors of upgraded GS.

Impact of Obesity on Long-Term Urinary Incontinence after Radical Prostatectomy: A Meta-Analysis.

Obesity is a known risk factor for prostate cancer progression and may contribute to poor treatment outcomes. However, little is known concerning the relationship between obesity (body mass index [BMI] ⩾ 30) and the urinary incontinence (UI) of patients after radical prostatectomy (RP). The goal of this study was to focus on the prevalence and duration of UI after RP with specific attention to the BMI. Subsequently, trials were identified in a literature search of PubMed, Embase, Cochrane Library, Web of Science, and Google Scholar using appropriate search terms. All comparative studies reporting BMI, study characteristics, and outcome data including the relationship between BMI and urinary incontinence data were included. Finally, four studies comprising 6 trials with 2890 participants were included. The results showed that obesity increased UI risk at 12 months in patients who underwent robotic-assisted laparoscopic radical prostatectomy (RLRP) (odds ratio [OR] 2.43, 95% confidence interval [CI] [1.21, 4.88], P = 0.01). When stratified by the surgical methods, the pooled results showed that obesity increased UI risk at 24 months in patients who underwent RLRP (OR 2.00, 95% CI [1.57, 2.56], P < 0.001). However, in patients who underwent laparoscopic radical prostatectomy (LRP), the pooled results showed that obesity does not increase UI risk at 24 months (OR 1.13, 95% CI [0.74, 1.72], P = 0.58). This is the first study to include obesity as the primary independent variable. Outcomes indicate that obesity (BMI ≥ 30) may increase the UI risk at 12 and 24 months after RLRP. Well-designed randomized controlled trials with strict control of confounders are needed to make results comparable.

Risk factors for bladder cancer recurrence survival in patients with upper-tract urothelial carcinoma.

PURPOSE: The aim of this work was to investigate the predictive factors for bladder cancer recurrence survival (BCRS) in patients with upper-tract urothelial carcinoma (UTUC). METHODS: We selected patients with UTUC who underwent segmental ureterectomy (Su) or nephroureterectomy (Nu) from 2004 to 2013 from the Surveillance, Epidemiology, and End Results (SEER) database. Patients with a history of intravesical therapy for bladder cancer and bladder cancer prior to the diagnosis of UTUC were excluded. We used Kaplan-Meier analysis, log-rank tests, and Cox proportional hazards model to compare overall survival, cancer-specific survival, and BCRS. RESULTS: In a cohort of 1,454 patients, 169 (11.6%) had low-grade tumors and 1,285 (88.4%) had high-grade tumors; 239 (16.4%) underwent Su and 1,215 (83.6%) underwent Nu. We found that T4 grade (hazard ratio [HR] = 6.216; 95% confidence interval [CI], 3.197-12.087) and ureteral tumors (HR = 1.764; 95% CI, 1.173-2.652) were predictors of shorter BCRS, whereas Nu (HR = 0.608; 95% CI, 0.388-0.953) predicted longer BCRS. Five-year BCRS rates were low-grade tumors: 94.1%, high-grade tumors: 85.4% (p = 0.038); plus Su: 82.9%, and Nu: 87.6% (p = 0.016). CONCLUSIONS: Use of Su should be more selective for high-grade tumors, as it correlates with shorter BCRS. Tumors located in the ureter are associated with shorter BCRS than those located in the renal pelvis.

Upregulation of Arp2 expression is associated with the prognosis and prediction of lymph node metastasis in bladder urothelial carcinoma.

OBJECTIVE: Upregulation of actin-related protein 2/3 complex subunit 2 (Arp2) is observed in various tumors, but its expression pattern has not been revealed in bladder urothelial carcinoma (BUC). The purpose of this study was to investigate the role of Arp2 expression in the pathological features and the prognosis as well as lymph node metastasis of BUC. METHODS: A total of 228 tissue specimens from BUC patients who underwent a radical cystectomy were collected. In addition, 40 lymph node specimens and 40 normal bladder tissue specimens as controls were used. All of the specimens were used to construct a tissue microarray and were subsequently subjected to immunohistochemical staining for Arp2 expression. Logistic and Cox regression analyses and Kaplan-Meier curve analysis were applied to analyze the relation between Arp2 expression and multiple clinical features of patients with BUC. RESULTS: Immunohistochemical staining observation showed that Arp2 was mainly expressed in the cytoplasm and nucleus of positive cells and that Arp2 expression in BUC was significantly higher than that in normal bladder tissues. Arp2 expression in BUC tissues was associated with tumor size, tumor multiplicity, tumor stage, tumor grade, and lymph node metastasis (P < 0.05). Logistic regression analysis demonstrated that Arp2 expression was an independent risk factor for lymph node metastasis of BUC (P < 0.05). Kaplan-Meier curve analysis showed that increased Arp2 expression was associated with a shortened recurrence-free survival (RFS) and overall survival (OS) of BUC patients after radical cystectomy (P < 0.05). CONCLUSION: These findings suggest that Arp2 is significantly upregulated in BUC tissues when compared with normal bladder tissues, and that Arp2 expression is an independent predictor for lymph node metastasis, RFS, and OS.

Structural Diversity and Biological Activities of Fungal Cyclic Peptides, Excluding Cyclodipeptides.

Cyclic peptides are cyclic compounds formed mainly by the amide bonds between either proteinogenic or non-proteinogenic amino acids. This review highlights the occurrence, structures and biological activities of fungal cyclic peptides (excluding cyclodipeptides, and peptides containing ester bonds in the core ring) reported until August 2017. About 293 cyclic peptides belonging to the groups of cyclic tri-, tetra-, penta-, hexa-, hepta-, octa-, nona-, deca-, undeca-, dodeca-, tetradeca-, and octadecapeptides as well as cyclic peptides containing ether bonds in the core ring have been isolated from fungi. They were mainly isolated from the genera Aspergillus, Penicillium, Fusarium, Acremonium and Amanita. Some of them were screened to have antimicrobial, antiviral, cytotoxic, phytotoxic, insecticidal, nematicidal, immunosuppressive and enzyme-inhibitory activities to show their potential applications. Some fungal cyclic peptides such as the echinocandins, pneumocandins and cyclosporin A have been developed as pharmaceuticals.

Clostridium Difficile Infection Worsen Outcome of Hospitalized Patients with Inflammatory Bowel Disease.

The prevalence of Clostridium difficile infection (CDI) in patients suffering from inflammatory bowel disease (IBD) has increased rapidly over the past several decades in North America and Europe. However, the exact global epidemiology remains unclear because of insufficient data from developing countries. A total of 646 hospitalized adult IBD patients were enrolled; and their fresh stool specimens were obtained and used for Clostridium difficile detection. The incidence of CDI in Crohn's disease (CD) patients (12.7%) was significantly lower than that in Ulcerative disease (UC) patients (19.3%). Among the toxin types, A(+)B(+) strain was the most common. Length of stay, hospitalization frequency and bowel surgery rate were significantly higher in the CDI than in the non-CDI group in CD or UC patients. More patients in CDI-CD group were still in active and even clinical moderate or severe CD stage than non-CDI-CD group after 2 years of following-up. Fistula, antibiotics and infliximab usage likely increased the CDI rate in CD patients, Infliximab treatment was considered a risk factor in UC patients. CDI is an exacerbating public health issue that may influence IBD course, increase expenditures, and delay the remission of IBD patients. IBD patients with CDI require urgent attention.

Ecological and morphological characteristics of parasitoids in Phauda flammans (Lepidoptera, Zygaenidae).

Phauda flammans Walker (Lepidoptera, Zygaenidae) is one of the notorious defoliators on Ficus spp. trees. In order to avoid environmental pollution, potential biological control agents for P. flammans need to be investigated instead of chemical control. Four species of insect parasitoids were identified from P. flammans, including three hymenopteran species (i.e., Gotra octocinctus, Apanteles sp. and Eurytoma verticillata) and one dipteran species (i.e., Exorista yunnanica). Parasitoid ratios of G. octocinctus, Apanteles sp., Eu. verticillata and Ex. yunnanica were 7.2%, 4.2%, 1.6% and 0.9%. The four species were all larval endoparasitoids of P. flammans larvae. Time of cocoon (pupa) to adult, life span, major axis of cocoon and body length of females were all longer compared to males for G. octocinctus, Apanteles sp. and Ex. yunnanica. Based on the parasitoid ratios, the most abundant parasitoid species was G. octocinctus.

Infliximab for the treatment of Crohn's disease: efficacy and safety in a Chinese single-center retrospective study.

BACKGROUND AND AIM: Information describing the efficacy and safety of infliximab (IFX) for Crohn's disease (CD) in China is limited. The aim of this study was to report response rates, relapse rates after IFX-induced remission, and adverse events, and identify factors associated with relapse in a cohort of CD patients treated with IFX in a Chinese center. PATIENTS AND METHODS: Clinical data of CD patients treated with two or more infusions of IFX from July 2010 to February 2013 at Nanfang Hospital (Guangzhou, China) were retrieved and analyzed retrospectively. Primary measurements were clinical response rate and relapse rate during 30 weeks of follow-up. Overall adverse events and risk factors related to IFX were evaluated. RESULTS: A total of 70 CD patients were finally included. The clinical response rate was 98.6% at week 2, 97% at week 6, 92.5% at week 14, 84.9% at week 22, and 84.1% at week 30. The remission rate was 55.7% at week 2, 94% at week 6, 92.5% at week 14, 82.7% at week 22, and 77.3% at week 30. Relapse began following the third infusion. The relapse rate was 3.23% at week 6, 6.12% at week 14, 10.53% at week 22, and 8.82% at week 30. Adverse events related to IFX occurred in 32.6% of patients; most were mild and transient. Univariate analysis showed that sex was a significant predictor of clinical relapse. CONCLUSION: IFX is an effective and safe treatment for CD in Chinese patients. Sex may be an independent predictor of relapse.

Rapid detection of ermB gene in Clostridium difficile by loop-mediated isothermal amplification.

Macrolide-lincosamide-streptogramin B resistance in Clostridium difficile is mostly due to the ermB resistance determinant. Here, we describe a sensitive and rapid molecular method to detect ermB in C. difficile to contribute to the wider epidemiological study. Five sets of loop-mediated isothermal amplification (LAMP) primers were designed and optimized for rapid detection of ermB. The specificity and sensitivity of the primers for ermB were detected, and the ermB LAMP assay was compared to conventional PCR with 80 clinical isolates of C. difficile. Real-time monitoring of turbidity and chromogenic reaction were used to determine negative and positive results. A total of 26 pathogenic bacterial strains of different species were found to be negative for ermB, which indicated the high specificity of the primers. ermB was detected in 78.8 % (63/80) of the clinical isolates by both LAMP and conventional PCR. The detection limit of LAMP was 36.1  pg DNA µl- 1 and its sensitivity was 10-fold greater than that of conventional PCR. This study is the first report regarding the development and application of the LAMP assay for detection of the ermB gene in C. difficile strains. The developed LAMP method is sensitive, specific and provides a user-friendly visual approach for the rapid detection of ermB-bearing C. difficile.

Advanced oxidation protein products decrease the expression of calcium transport channels in small intestinal epithelium via the p44/42 MAPK signaling pathway.

Advanced oxidation protein products (AOPPs), novel protein markers of oxidative damage, accumulate in the plasma of patients with inflammatory bowel disease (IBD). Osteoporosis, which is closely related to the regulation of intestinal calcium transport channels (CTCs), is a prevalent extraintestinal complication of IBD and is associated with oxidative stress. However, the underlying mechanisms are unknown. The present study aimed to verify whether AOPPs inhibit CTCs in the small intestinal epithelium and to identify the underlying mechanisms that may contribute to IBD-associated osteoporosis. Normal Sprague-Dawley rats were treated with AOPP-modified rat serum albumin. The calcium ion level in serum was not significantly altered, while the duodenal expression of CTCs (e.g. transient receptor potential vanilloid [TRPV6], calbindin-D9k [CaBP-D9k], plasma membrane Ca(2+)-ATPase 1 [PMCA1], and Na(+)/Ca(2+) exchanger 1 [NCX1]) were decreased. In contrast, the levels of the related hormones that regulate calcium absorption including parathyroid hormone (PTH), 25-(OH)D3, and 1,25-(OH)2D3 were increased, although the trend toward an increase in PTH levels was not significant. In order to further investigate the effects of AOPP exposure, we also evaluated the expression of CTCs (including the voltage-dependent L-type calcium channel [CaV1.3], TRPV6, CaBP-D9k, PMCA1, and NCX1) in cultured human colorectal adenocarcinoma cells (Caco-2). The expression levels of total CTC protein and mRNA, except for CaV1.3, were significantly down-regulated in a concentration- and time-dependent manner. Moreover, phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) was observed in vivo and in vitro. The p44/42 inhibitor U0126 reversed the down-regulation of CTCs induced by AOPPs in the Caco-2 monolayer. Our results indicate that AOPPs down-regulate the expression of CTCs through p44/42 MAPK signaling mechanisms in the small intestinal epithelium. These data provide new insights regarding the molecular basis of AOPP-induced reductions in intestinal CTCs, and are relevant to understanding the mechanisms of IBD-associated osteoporosis. Further studies are needed to explore these mechanisms in greater detail.

Divergent evolution of the chloroplast small heat shock protein gene in the genera Rhododendron (Ericaceae) and Machilus (Lauraceae).

BACKGROUND AND AIMS: Evolutionary and ecological roles of the chloroplast small heat shock protein (CPsHSP) have been emphasized based on variations in protein contents; however, DNA sequence variations related to the evolutionary and ecological roles of this gene have not been investigated. In the present study, a basal angiosperm, Machilus, together with the eudicot Rhododendron were used to illustrate the evolutionary dynamics of gene divergence in CPsHSPs. METHODS: Degenerate primers were used to amplify CPsHSP-related sequences from 16 Rhododendron and eight Machilus species that occur in Taiwan. Manual DNA sequence alignment was carried out according to the deduced amino acid sequence alignment performed by CLUSTAL X. A neighbour-joining tree was generated in MEGA using conceptual translated amino acid sequences from consensus sequences of cloned CPsHSP genes from eight Machilus and 16 Rhododendron species as well as amino acid sequences of CPsHSPs from five monocots and seven other eudicots acquired from GenBank. CPsHSP amino acid sequences of Funaria hygrometrica were used as the outgroups. The aligned DNA and amino acid sequences were used to estimate several parameters of sequence divergence using the MEGA program. Separate Bayesian inference of DNA sequences of Rhododendron and Machilus species was analysed and the resulting gene trees were used for detection of putative positively selected amino acid sites by the Codeml program implemented in the PAML package. Mean hydrophobicity profile analysis was performed with representative amino acid sequences for both Rhododendron and Machilus species by the Bioedit program. The computer program SplitTester was used to examine whether CPsHSPs of Rhododendron lineages and duplicate copies of the Machilus CPsHSPs have evolved functional divergence based on the hydrophobicity distance matrix. KEY RESULTS: Only one copy of the CPsHSP was found in Rhododendron. However, a higher evolutionary rate of amino acid substitutions in the Hymenanthes lineage of Rhododendron was inferred. Two positively selected amino acid sites may have resulted in higher hydrophobicity in the region of the alpha-crystallin domain (ACD) of the CPsHSP. By contrast, the basal angiosperm, Machilus, possessed duplicate copies of the CPsHSP, which also differed in their evolutionary rates of amino acid substitutions. However, no apparent relationship of ecological relevance toward the positively selected amino acid sites was found in Machilus. CONCLUSIONS: Divergent evolution was found for both Rhododendron lineages and the paralogues of CPsHSP in Machilus that were directed to the shift in hydrophobicity in the ACD and/or methionine-rich region, which might have played important roles in molecular chaperone activity.