Curcumin Inhibits Lipopolysaccharide-Induced Mucin 5AC Hypersecretion and Airway Inflammation via Nuclear Factor Erythroid 2-Related Factor 2 / 中华医学杂志(英文版)

<p><b>Background</b>Excess mucus production is an important pathophysiological feature of chronic inflammatory airway diseases. Effective therapies are currently lacking. The aim of the study was to evaluate the effects of curcumin (CUR) on lipopolysaccharide (LPS)-induced mucus secretion and inflammation, and explored the underlying mechanism in vivo and in vitro.</p><p><b>Methods</b>For the in vitro study, human bronchial epithelial (NCI-H292) cells were pretreated with CUR or vehicle for 30 min, and then exposed to LPS for 24 h. Next, nuclear factor erythroid 2-related factor 2 (Nrf2) was knocked down with Nrf2 small interfering RNA (siRNA) to confirm the specific role of Nrf2 in mucin regulation of CUR in NCI-H292 cells. In vivo, C57BL/6 mice were randomly assigned to three groups (n = 7 for each group): control group, LPS group, and LPS + CUR group. Mice in LPS and LPS + CUR group were injected with saline or CUR (50 mg/kg) intraperitoneally 2 h before intratracheal instillation with LPS (100 μg/ml) for 7 days. Cell lysate and lung tissue were obtained to measured Mucin 5AC (MUC5AC) and Nrf2 mRNA and protein expression by a real-time polymerase chain reaction and Western blotting. Bronchoalveolar lavage fluid (BALF) was collected to enumerate total cells and neutrophils. Histopathological changes of the lung were observed. Data were analyzed by one-way analysis of variance. Student's t-test was used when two groups were compared.</p><p><b>Results</b>CUR significantly decreased the expression of MUC5AC mRNA and protein in NCI-H292 cells exposed to LPS. This effect was dose dependent (2.424 ± 0.318 vs. 7.169 ± 1.785, t = 4.534, and 1.060 ± 0.197 vs. 2.340 ± 0.209, t = 7.716; both P < 0.05, respectively) and accompanied by increased mRNA and protein expression of Nrf2 (1.952 ± 0.340 vs. 1.142 ± 0.176, t = -3.661, and 2.010 ± 0.209 vs. 1.089 ± 0.132, t = -6.453; both P < 0.05, respectively). Furthermore, knockdown of Nrf2 with siRNA increased MUC5AC mRNA expression by 47.7%, compared with levels observed in the siRNA-negative group (6.845 ± 1.478 vs. 3.391 ± 0.517, t = -3.821, P < 0.05). Knockdown of Nrf2 with siRNA also markedly increased MUC5AC protein expression in NCI-H292 cells. CUR also significantly decreased LPS-induced mRNA and protein expression of MUC5AC in mouse lung (1.672 ± 0.721 vs. 5.961 ± 2.452, t = 2.906, and 0.480 ± 0.191 vs. 2.290 ± 0.834, t = 3.665, respectively; both P < 0.05). Alcian blue/periodic acid-Schiff staining also showed that CUR suppressed mucin production. Compared with the LPS group, the numbers of inflammatory cells (247 ± 30 vs. 334 ± 24, t = 3.901, P < 0.05) and neutrophils (185 ± 22 vs. 246 ± 20, t = 3.566, P < 0.05) in BALF decreased in the LPS + CUR group, as well as reduced inflammatory cell infiltration in lung tissue.</p><p><b>Conclusion</b>CUR inhibits LPS-induced airway mucus hypersecretion and inflammation through activation of Nrf2 possibly.</p>

Use of subsequent PET/CT in diffuse large B-cell lymphoma patients in complete remission following primary therapy / 癌症

Interim 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (I-PET/CT) is a powerful tool for monitoring the response to therapy in diffuse large B-cell lymphoma (DLBCL). This retrospective study aimed to determine when and how to use I-PET/CT in DLBCL. A total of 197 patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) were enrolled between October 2005 and July 2011; PET/CT was performed at the time of diagnosis (PET/CT0), after 2 and 4 cycles of chemotherapy (PET/CT2 and PET/CT4, respectively), and at the end of treatment (F-PET/CT). According to the International Harmonization Project for Response Criteria in Lymphoma, 110 patients had negative PET/CT2 scans, and 87 had positive PET/CT2 scans. The PET/CT2-negative patients had significantly higher 3-year progression-free survival rate (75.8% vs. 38.2%) and 3-year overall survival rate (93.5% vs. 55.6%) than PET/CT2-positive patients. All PET/CT2-negative patients remained negative at PET/CT4, but 3 were positive at F-PET/CT. Among the 87 PET/CT2-positive patients, 57 remained positive at F-PET/CT, and 32 progressed during chemotherapy (15 at PET/CT4 and 17 at F-PET/CT). Comparing PET/CT4 with PET/CT0, 7 patients exhibited progression, and 8 achieved partial remission. Comparing F-PET/CT with PET/CT0, 10 patients exhibited progression, and 7 achieved partial remission. In conclusion, our results indicate that I-PET/CT should be performed after 2 rather than 4 cycles of immunochemotherapy in DLBCL patients. There is a limited role for subsequent PET/CT in the detection of relapse in PET/CT2-negative patients, but repeat PET/CT is required if the PET/CT2 findings are positive.

Qualitative visual trichotomous assessment improves the value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in predicting the prognosis of diffuse large B-cell lymphoma / 癌症

<p><b>INTRODUCTION</b>Fluorine-18 fluorodeoxyglucose (18 F-FDG) positron emission tomography/computed tomography (PET/CT) is a powerful tool for monitoring the response of diffuse large B-cell lymphoma (DLBCL) to therapy, but the criteria to interpret PET/CT results remain under debate. We investigated the value of post-treatment PET/CT in predicting the prognosis of DLBCL patients when interpreted according to qualitative visual trichotomous assessment (QVTA) criteria compared with the Deauville criteria.</p><p><b>METHODS</b>In this retrospective study, final PET/CT scans of DLBCL patients treated with rituximab-based regimens between October 2005 and November 2010 were interpreted using the Deauville and QVTA criteria. Survival curves were estimated using Kaplan-Meier analysis and compared using the log-rank test.</p><p><b>RESULTS</b>A total of 253 patients were enrolled. The interpretation according to the Deauville criteria revealed that 181 patients had negative PET/CT scan results and 72 had positive results. The 3 year overall survival (OS) rate was significantly higher in patients with negative scan results than in those with positive results (91.6% vs. 57.5%, P<0.001). The 72 patients with positive scan results according to the Deauville criteria were divided into two groups by the interpretation according to the QVTA criteria: 29 had indeterminate results, and 43 had positive results. The 3 year OS rate was significantly higher in patients with indeterminate scan results than in those with positive results (91.2% vs. 33.5%, P<0.001) but was similar between patients with negative and indeterminate scan results (91.6% vs. 91.2%, P=0.921).</p><p><b>CONCLUSIONS</b>Compared with the Deauville criteria, using the QVTA criteria for interpreting post-treatment PET/CT scans of DLBCL patients is likely to reduce the number of false positive results. The QVTA criteria are feasible for therapeutic outcome evaluation and can be used to guide risk-adapted therapy.</p>

Clinical impact of 18F-FDG PET/CT on suspected cervical cancer recurrence / 中华核医学杂志

Objective To evaluate the clinical impact of 18F-FDG PET/CT on patients with suspected cervical cancer recurrence. Methods Fifty-one cervical cancer patients, clinically suspected to have tumor recurrence during follow-up, underwent 18F-FDG PET/CT examination. 18 F-FDG PET/CT results were compared with those of conventional images, as referred to histopathology or clinical follow-up. Impacts of 18F-FDG PET/CT were evaluated based on documented changes of clinical management. Results In total, 43 patients were found to have positive lesions by 18F-FDG PET/CT, in which 40 were true recurrence,but 2 were pelvic abscess and 1 was radiation enterocolitis. Other 8 patients were found negative by 18F-FDG PET/CT and confirmed by pathology or follow-up. In patient-based analyses, the sensitivity, specificity, and accuracy of 18F-FDG PET/CT for the detection of tumor recurrence were 100% (40/40), 72. 73% (8/11),and 94.12% (48/51) respectively. In 7 patients, the clinical management was changed due to 18F-FDG PET/CT findings. Conclusion 18F-FDG PET/CT is an efficient tool for determining the recurrence of cervical cancer and instructing the clinical management.

Allergen micro-array detection of specific IgE-reactivity in Chinese allergy patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Allergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among selected Chinese allergy patients.</p><p><b>METHODS</b>The study was conducted using patient sera from the cities of Guangzhou, Nanjing, Chengdu and Shenyang. In total 100 sera with Dermatophagoides pteronyssinus (Der p) specific IgE-levels higher than 50 kU/L were selected for testing against 103 individual allergens.</p><p><b>RESULTS</b>Among 100 selected patients, 95% showed IgE-reactivity towards house-dust mite allergens Dermatophagoides farinae (Der f) 1, Der f 2 and Der p 2 and 94% were IgE positive against Der p 1, and 60% of sera contained IgE reacting against allergen Euroglyphus maynei (Eur m) 2. IgE against cat allergen, Felisdomesticus (Fel d) 1, was seen in 20%. Only 2% showed specific IgE-reactivity to Der p 10, a panallergen belonging to the tropomyosin family. Serum IgE-reactivity towards other allergens was in general low. IgE-reactivity against pollen allergens showed geographic differences.</p><p><b>CONCLUSIONS</b>This study clearly confirms that group 1 and group 2 are major allergens of house dust mites. These selected house-dust mite allergy patients are close to being mono-sensitized. Der p 10 is not an important allergen for cross-reactivity. Specific IgE-sensitization towards pollen allergens is low in southern China compared to other regions. The prevalence of food and stinging insect allergens known to give rise to IgE-mediated cross-reactivity is 2% or less.</p>

Novel GATA4 mutations identified in patients with congenital atrial septal defects / 中华心血管病杂志

<p><b>OBJECTIVE</b>To identify the genetic defects in patients with congenital atrial septal defects (ASD).</p><p><b>METHODS</b>The clinical data and blood samples from 180 unrelated subjects with congenital ASD were collected and evaluated. Two hundred healthy individuals served as controls. The coding exons and the flanking introns of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with the sequences publicized in GenBank by the aid of programme BLAST to identify the sequence variations. Clustal W software was applied for analysis of the conservation of altered amino acids.</p><p><b>RESULTS</b>Two novel heterozygous missense GATA4 mutations were identified in 2 out of 180 ASD patients. Namely, the triplet substitutions of GTC for GGC at codon 21 and TCG for CCG at codon 87 were detected, predicting the conversions of glycine into valine at amino acid residue 21 (G21V) and proline into serine at amino acid residue 87 (P87S). None of the two mutations were detected in 200 healthy controls. Across-species alignment of GATA4 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, a single nucleotide polymorphism c.99G>T was observed. However, the polymorphic frequency distribution in ASD cases was similar with that in healthy controls (for genotype GT, χ(2) = 0.7556, P = 0.3847; for allele T, χ(2) = 0.7235, P = 0.3950).</p><p><b>CONCLUSIONS</b>Two novel mutations of GATA4 gene are identified in two unrelated ASD patients. This finding provides new insight into the molecular etiology responsible for ASD.</p>

A novel GATA4 mutation leading to congenital ventricular septal defect / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the GATA4 gene mutation of congenital ventricular septal defect (VSD) and study the molecular mechanism of a novel mutation.</p><p><b>METHODS</b>The clinical data and blood samples from 185 unrelated subjects with congenital VSD were collected and evaluated together with 200 healthy individuals. The coding exons and the flanking intron regions of the GATA4 gene were amplified by PCR and sequenced using the di-deoxynucleotide chain termination approach. The GATA4 gene was cloned and the corresponding mutant was acquired by site directed mutagenesis. The recombinant plasmid expressing GATA4 and the reporter vector expressing enhanced green fluorescence protein (EGFP) driven by the promoter of atrial natrium peptide (ANP) gene were transfected into HeLa cells with Lipofectamine. The effect of mutated GATA4 gene on the transcriptional activity of encoded transcriptional factor was analyzed by reverse transcription (RT)-PCR.</p><p><b>RESULTS</b>A novel heterozygous missense GATA4 mutation, c.191G>A was identified in 1 VSD patient. The mutation leads to glycine to glutamic acid change at amino acid residue 64 (G64E) in the GATA4 protein. Functional analysis showed that GATA4 G64E mutation decreased the transcriptional activity of GATA4 transcriptional factor.</p><p><b>CONCLUSION</b>A novel heterozygous missense GATA4 mutation, G64E, was identified in 1 VSD patient. The mutation might cause VSD by impairing the transcriptional activity of GATA4 transcriptional factor.</p>

Mutation of NKX2-5 gene in patients with atrial septal defect / 中华儿科杂志

<p><b>OBJECTIVE</b>The purpose of this investigation was to identify the novel genetic mutations in patients with a congenital atrial septal defect (ASD).</p><p><b>METHODS</b>Clinical data and blood specimens from a total of 12 unrelated ASD pedigrees and a cohort of 168 unrelated subjects with sporadic ASD were collected and evaluated in contrast to 200 healthy individuals. The whole exons and partial flanking introns of NKX2-5 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with those publicized in GenBank by the aid of programme BLAST to identify the sequence variations. The software ClustalW was applied for analysis of the conservative of the altered amino acids.</p><p><b>RESULTS</b>A novel heterozygous mutation of NKX2-5 gene, i.e., a substitution of thymine for cytosine at nucleotide 536, predicting the conversion of serine into phenylalanine at amino acid residue 179, was identified initially in a proband. The same missense mutation was thereafter detected in other 3 affected members of the identical family but neither in the healthy members of the kindred nor in the 200 normal controls. A cross-species alignment of the protein sequences encoded by NKX2-5 gene displayed that the mutated amino acid was highly conserved evolutionarily. No mutation of NKX2-5 gene was observed in the other 11 ASD pedigrees or in 168 patients with sporadic ASD. Additionally, a common synonymous single nucleotide polymorphism, a transition of adenine (A) into guanine (G) at nucleotide 63, was found in NKX2-5 gene. However, there were no significant differences in the prevalence of alleles A and G between ASD patients and healthy controls (chi(2) = 2.8641, P = 0.0906).</p><p><b>CONCLUSION</b>A novel mutation of NKX2-5 gene identified in an ASD family suggests that mutated NKX2-5 gene is responsible for familial ASD.</p>

Diagnostic value of FDG-PET in the detection of bone marrow involvement in patients with diffuse large B-cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the diagnostic value of bone marrow (BM) involvement detected by F-18-fluorodeoxyglucose-positron emission tomography/computed tomography ((18)F-FDG PET/CT) in patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL).</p><p><b>METHODS</b>The study population comprised 81 consecutive patients with newly diagnosed DLBCL. All patients had both (18)F-FDG PET/CT and standard BM biopsy at iliac crest. In 9 patients, additional direct BM biopsy at FDG-avid bone lesion was performed.</p><p><b>RESULTS</b>Among all 81 cases, 23 were diagnosed as BM involvement by PET/CT. Of the 23 positive cases 17 were confirmed by biopsy. However, only 11 cases were diagnosed by merely bilateral iliac crest biopsy. In patients in early stage of disease (18)F-FDG PET/CT had the same results as bilateral iliac crest biopsy.</p><p><b>CONCLUSION</b>(18)F-FDG PET/CT is superior to standard BM biopsy in detecting BM involvement in newly diagnosed DLBCL patients. In patients with FDG-avid bone lesions, direct PET/CT-guided bone biopsy seems to be more accurate than standard BM biopsy.</p>

Isolation and Identification of BLN-2 Producing Poly-?-Glutamic Acid and Studies on Its Solid-state Fermentation / 微生物学通报

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.