A simple approach to the determination of three curcuminoids with similar chemical structures in different substrates.

The determination of curcuminoids in mixtures is more difficult due to their similar chemical structures as well as serious interferences, thus the complex pretreatments of samples and the optimization of experimental conditions are often required. Here, owing to the mathematical separation of chemical signals by Tchebichef image moments, a simple and effective approach to the simultaneous quantitative analysis was proposed, and applied to the determination of the three curcuminoids in turmeric and curry based on their raw fluorescence 3D spectra. For the established linear models, the leave-one-out correlation coefficients (R loo-cv) were more than 0.9816 within the linear ranges, and the predictive correlation coefficients (R p) for the external independent samples were more than 0.9897. The intra- and inter-day precision (less than 6.82%, RSD), average spiked recovery (89.9% ~ 100.8%), LOD (less than 0.07 µg/mL) and LOQ (less than 0.23 µg/mL) suggest that the proposed approach is accurate and reliable. Compared with N-PLS and MCR-ALS methods, our method can obtain more satisfactory results. This study provides a convenient pathway for the rapid analysis of multi-target components with similar chemical structures in mixture of different substrates.

Novel Approach to the Analysis of Chemical Third-Order Data.

For the more complex samples, chemical higher-order data can be collected from various information sources, which become the necessary foundation of accurate analysis. In this article, the Tchebichef cubic moment (TCM) was developed for the analysis of chemical third-order data for the first time. Then, the proposed TCM approach was applied to the fluorescence excitation-emission time data for the analysis of adrenaline and noradrenaline in urinary samples (Data I) and the data fusion of the excitation-emission matrix (EEM), NMR, and liquid chromatography-mass spectrometry (LC-MS) spectra for the determination of the five target components (Data II). For Data I, all of the cross-validation correlation coefficients (Rcv2) of the obtained linear models on the calibration set were more than 0.9937 and the prediction root-mean-square errors (RMSEp) of the external independent test samples were less than 0.0250 µM. For Data II, all of the Rcv2 were higher than 0.9846 and RMSEp were less than 0.2267 µM. Compared with several conventional methods, the proposed method was more convenient and accurate. This study provides another effective approach to the analysis of complex samples based on their chemical third-order data.

Simultaneous quantification of multiple endogenous biothiols in cancer cells based on a multi-signal fluorescent probe.

Although a large number of fluorescent probes have been developed, the simultaneous quantitative analysis of intracellular thiols is still difficult due to the spectral overlap and the complexity of the intracellular environment. In this study, a multi-signal fluorescent probe was employed for the simultaneous quantification of intracellular glutathione (GSH), cysteine (Cys) and homocysteine (Hcy). As the feature variables of the target components, the Tchebichef image moments (TMs) calculated from the grayscale images of the 3D fluorescence spectra were used to establish the quantitative linear models by stepwise regression. The intra-day and inter-day precisions of the proposed method were less than 5.6% and 8.7%, respectively. The recoveries ranged from 97.0% to 105.9%. In addition, the proposed approach was applied to the simultaneous quantitative determination of Cys, GSH and Hcy in the MCF-10A cell (a type of normal cell) and MDA-MB-231 cancer cell. The obtained results indicated that the concentrations of the three thiols in the cancer cell were higher than those in the normal cell. This study not only provides an effective approach for the quantification of multi-target bio-molecules in complicated intracellular environments, but also further extends the applications of multi-signal fluorescent probes, which will promote the design of new multi-signal fluorescent probes.

Applications of Discrete Shmaliy Moments on the Quantitative Analysis of Multitarget Compounds Based on the Chemical Spectra.

To extract the features in first-order or second-order signals, the two kinds of discrete Shmaliy moment (DSM) methods were proposed and applied to the quantitative analysis of multitarget compounds in complexes based on the UV-vis and high-performance liquid chromatography with pulsed amperometric detector (HPLC-PAD) spectra of samples for the first time. All the statistical parameters demonstrated that the obtained models were accurate and the established analytical methods were reliable, even in the presence of a different degree of overlapping signals as well as various interferences. Compared with Tchebichef moment (TM) and other classical methods such as multivariate curve resolution-alternating least-squares (MCR-ALS), partial least-squares (PLS) regression, and N-way partial least-squares (N-PLS), the proposed methods are more convenient and efficient, which not only provides another suitable tool for the quantitative analysis of multitarget components in complex samples but also extends the application of moment invariants in chemical signal analyses.

Tchebichef image moment approach to the prediction of protein secondary structures based on circular dichroism.

Circular dichroism (CD) spectroscopy is a widely used technique for the evaluation of protein secondary structures that has a significant impact for the understanding of molecular biology. However, the quantitative analysis of protein secondary structures based on CD spectra is still a hard work due to the serious overlap of the spectra corresponding to different structural motifs. Here, Tchebichef image moment (TM) approach is introduced for the first time, which can effectively extract the chemical features in CD spectra for the quantitative analysis of protein secondary structures. The proposed approach was applied to analyze reference set and the obtained results were evaluated by the strict statistical parameters such as correlation coefficient, cross-validation correlation coefficient and root mean squared error. Compared with several specialized prediction methods, TM approach provided satisfactory results, especially for turns and unordered structures. Our study indicates that TM approach can be regarded as a feasible tool for the analysis of the secondary structures of proteins based on CD spectra. An available TMs package is provided and can be used directly for secondary structures prediction.

Two microRNA panels to discriminate three subtypes of lung carcinoma in bronchial brushing specimens.

RATIONALE: Effective treatment for lung cancer requires accuracy in subclassification of carcinoma subtypes. OBJECTIVES: To identify microRNAs in bronchial brushing specimens for discriminating small cell lung cancer (SCLC) from non-small cell lung cancer (NSCLC) and for further differentiating squamous cell carcinoma (SQ) from adenocarcinoma (AC). METHODS: Microarrays were used to screen 723 microRNAs in laser-captured, microdissected cancer cells from 82 snap-frozen surgical lung specimens. Quantitative reverse-transcriptase polymerase chain reaction was performed on 153 macrodissected formalin-fixed, paraffin-embedded (FFPE) surgical lung specimens to evaluate seven microRNA candidates discovered from microarrays. Two microRNA panels were constructed on the basis of a training cohort (n = 85) and validated using an independent cohort (n = 68). The microRNA panels were applied as differentiators of SCLC from NSCLC and of SQ from AC in 207 bronchial brushing specimens. MEASUREMENTS AND MAIN RESULTS: Two microRNA panels yielded high diagnostic accuracy in discriminating SCLC from NSCLC (miR-29a and miR-375; area under the curve [AUC], 0.991 and 0.982 for training and validation data set, respectively) and in differentiating SQ from AC (miR-205 and miR-34a; AUC, 0.977 and 0.982 for training and validation data set, respectively) in FFPE surgical lung specimens. Moreover, the microRNA panels accurately differentiated SCLC from NSCLC (AUC, 0.947) and SQ from AC (AUC, 0.962) in bronchial brushing specimens. CONCLUSIONS: We found two microRNA panels that accurately discriminated between the three subtypes of lung carcinoma in bronchial brushing specimens. The identified microRNA panels may have considerable clinical value in differential diagnosis and optimizing treatment strategies based on lung cancer subtypes.

Cholesteryl ester transfer protein TaqIB polymorphism and its association with serum lipid levels and longevity in Chinese Bama Zhuang population.

BACKGROUND: TaqIB polymorphism in the cholesteryl ester transfer protein (CETP) gene has been reported to be associated with serum high-density lipoprotein cholesterol (HDL-C) levels and longevity in several populations, but controversial results also arose probably due to racial/ethnic diversity. Bama is a remote and mountainous county located in the northwest of Guangxi, People's Republic of China, which has been well known for its longevity for centuries. The current study was to investigate the possible association of CETP TaqIB polymorphism with serum lipid levels and longevity in the Bama Zhuang population. METHODS: The CETP TaqIB genotypes were determined by polymerase chain reaction and restriction fragment length polymorphism in 523 long-lived inhabitants (long-lived group, LG; aged 90-107 years) and 498 healthy controls without longevity family history (non-long-lived group, non-LG; aged 40-69 years) residing in Bama County. RESULTS: The levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were higher but TG, HDL-C/LDL-C ratio and the prevalence of dyslipidemia were lower in LG than in non-LG (P < 0.001 for all). There were no differences in the allelic and genotypic frequencies between the two groups (P > 0.05). Serum HDL-C levels and HDL-C/LDL-C ratio in LG were different among the genotypes (P < 0.01 for each), the subjects with B2B2 and B1B2 genotyes had higher HDL-C levels and HDL-C/LDL-C ratio than the subjects with B1B1genotye, whereas the levels of TC and HDL-C in non-LG were different among/between the genotypes (P < 0.01 for each), the B2 allele carriers had lower TC and higher HDL-C levels than the B2 allele noncarriers. Serum TG and HDL-C levels and HDL-C/LDL-C ratio were correlated with genotypes in LG, whereas serum TC and HDL-C levels were associated with genotypes in non-LG (P < 0.05-0.001). CONCLUSIONS: The association of CETP TaqIB polymorphism and serum lipid profiles is different between LG and non-LG in the Chinese Bama Zhuang population. CETP TaqIB polymorphism might be one of the longevity-related genetic factors in this population.

Microsomal triglyceride transfer protein gene -493G/T polymorphism and its association with serum lipid levels in Bama Zhuang long-living families in China.

BACKGROUND: The -493G/T polymorphism in the microsomal triglyceride transfer protein (MTP) gene is associated with lower serum low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG) levels and longevity in several populations, but the results are inconsistent in different racial/ethnic groups. The current study was to investigate the plausible association of MTP -493G/T polymorphism with serum lipid levels and longevity in Zhuang long-lived families residing in Bama area, a famous home of longevity in Guangxi, China. METHODS: The MTP -493G/T was genotyped by PCR-restriction fragment length polymorphism in 391 Bama Zhuang long-lived families (BLF, n = 1467, age 56.60 ± 29.43 years) and four control groups recruited from Bama and out-of-Bama area with or without a familial history of exceptional longevity: Bama non-long-lived families (BNLF, n = 586, age 44.81 ± 26.83 years), Bama non-Zhuang long-lived families (BNZLF, n = 444, age 52.09 ± 31.91 years), Pingguo long-lived families (PLF, n = 658, age 50.83 ± 30.30 years), and Pingguo non-long-lived families (PNLF, n = 539, age 38.74 ± 24.69 years). Correlation analyses between genotypes and serum lipid levels and longevity were then performed. RESULTS: No particularly favorable lipoprotein and clinical phenotypes were seen in BLF as compared to general families in the same area. Instead, the levels of total cholesterol (TC), TG, LDL-C, and the prevalence of dyslipidemia were significantly higher in the three Bama families as compared to the two non-Bama families (P < 0.01 for all). There were no differences in the allelic and genotypic frequencies among the tested cohorts (P > 0.05 for all), but the TT genotype tended to enrich in the three long-lived cohorts from both areas. In addition, the individuals harboring TT genotype exhibited lower LDL-C and TC levels in the overall populations and Bama populations with a region- and sex-specific pattern. Multiple linear regression analyses unraveled that LDL-C levels were correlated with genotypes in Bama combined population, BNLF, and the total population (P < 0.05 for each) but not in Pingguo populations; TC and HDL-C levels were correlated with genotypes in Bama combined population and BLF, respectively (P < 0.05 for each). CONCLUSIONS: MTP -493G/T polymorphism may play an important role in fashioning the serum lipid profiles of Bama populations, despite no direct association between MTP -493G/T and longevity was detected.

Primary pulmonary mucoepidermoid carcinoma: an analysis of 21 cases.

BACKGROUND: The optimal treatment for pulmonary mucoepidermoid carcinoma (MEC), a rare type of tumor, has not been established yet. This study analyzed the survival of pulmonary MEC patients and attempted to find clues for optimal treatment. METHODS: A total of 21 patients with pulmonary MEC from November 2004 to January 2011 were included in the investigation. Immunohistochemistry, epidermal growth factor receptor (EGFR) mutation, and survival were retrospectively studied. RESULTS: Among the 21 pulmonary MEC patients, 17 were diagnosed with low-grade malignancy and 4 with high-grade malignancy through pathological examination. The prognosis was found to be poor in the presence of lymph nodes. The expression rates of EGFR and HER2 were 28.6% and 0%, respectively, which correlated with neither grade nor prognosis. The mutation rate of EGFR was 0. Log-rank test results indicated that age, grade, lymph node metastasis, and tumor-node-metastasis stage were prognostic factors. CONCLUSION: Age, grade, lymph node metastasis and tumor-node-metastasis stage correlate with the survival of pulmonary MEC patients. TRIAL REGISTRATION: This study was approved and registered by the Ethics Committee of Zhongshan Hospital. Written informed consent was obtained from all participants prior to treatment.

[Staging and histologic grading of gastrointestinal stromal tumors].

OBJECTIVE: To investigate the clinical stage and histological grade of gastrointestinal stromal tumors. METHODS: Twelve clinical and pathological parameters were assessed in 613 patients with follow-up information. These parameters were classified into two gross spread parameters including liver metastasis and peritoneal dissemination, five microscopic spread parameters including lymph node metastasis, vascular, fat, nerve and mucosal infiltration, and five histological parameters including mitotic count ≥ 10 per 50 high-power fields, muscularis propria infiltration, coagulative necrosis, perivascular pattern and severe nuclear atypia. RESULTS: The accumulated 5-year disease-free survival (DFS) and overall survival (OS) of 293 patients without any of these predictive parameters of malignancy were 99.3% and 100.0%, respectively. They were regarded as nonmalignant and further evaluations on the stage and grade of these tumors were not performed. At least one and at most seven predictive parameters of malignancy were identified in 320 patients. For these patients, the accumulated 5-year DFS and OS rates were 43.9% (mean 6.7 years) and 59.7% (mean 9.3 years), respectively. The DFS showed significant difference between patients with and without gross spread (P < 0.01), with and without microscopic spread (P = 0.001). DFS and OS were associated with the number of predictive parameters of malignancy in patients without gross spread (P < 0.01 for both DFS and OS), but not in patients with gross spread (P = 0.882 and 0.441, respectively). CONCLUSIONS: Malignant GIST could be divided into clinical stages I and II based on the absence and presence of gross spread, respectively. The degree of malignancy of patients in clinical stage I could be graded according to the number of predictive parameters of malignancy. Patients in clinical stage II were of the highest degree of malignancy regardless of the number of parameters. The staging and grading of gastrointestinal stromal tumors in this study are strongly associated with prognosis.

[Detection of KRAS gene mutation and its clinical significance in colorectal adenocarcinoma].

OBJECTIVE: To explore the clinical significance of KRAS mutation detection in colorectal adenocarcinoma. METHODS: Paraffin-embedded tissue specimens were obtained from 440 patients with colorectal adenocarcinoma. The genomic DNA was extracted. Mutations of exon 2 of KRAS gene were examined by PCR and direct sequencing. RESULTS: Somatic mutations of KRAS gene were identified in 146 cases, with the mutation rate of 33.2% (146/440). Among these 146 patients, KRAS mutation involved codon 12 in 118 patients, including 35G > A (Gly12Asp, 62 cases), 35G > T (Gly12Val, 35 cases), 34G > T (Gly12Cys, 9 cases), 34G > A (Gly12Ser, 6 cases), 35G > C (Gly12Ala, 5 cases), and 34G > C (Gly12Arg, 1 case); in 27 patients the mutation involved codon 13, including 38G > A (Gly13Asp, 25 cases), 38G > C (Gly13 Val, 1 case) and 37G > T (Gly13 Cys, 1 case); and in one patient, the mutation involved codon 14 with 40G > A (Val14Ile). The status of KRAS or codon 12 mutations in colorectal adenocarcinoma was related to patients' gender (P = 0.021 and P = 0.030, respectively), and this significant correlation to females was conserved in clinical stage III (P = 0.007 and P = 0.003, respectively), but not in stages I, II, and IV. The status of KRAS or codon 12 mutations was also related to tumor stage. Between stage II and stage IV, the mutation rate of KRAS and codon 12 showed significant difference (P = 0.028 and 0.034, respectively). Between stage III and stage IV, only the codon 12 mutation rate showed significant difference (P = 0.011). Codon 13 mutation was not related to tumor stage. CONCLUSION: About one third of patients with colorectal adenocarcinoma have KRAS gene mutation, which might be related to patients' gender; and could be consistently detected by PCR and direct sequencing.

The association between adult asthma and superoxide dismutase and catalase gene activity.

BACKGROUND: Adult asthma is caused by interaction effects of multiple genetic and environmental factors. Some studies have suggested that antioxidant enzyme activity and gene polymorphisms may play important roles in the context of asthma. Therefore, our study objectives were to investigate the association between asthma, antioxidant activities and the polymorphisms of manganese superoxide dismutase (Mn-SOD) or catalase (CAT). MATERIALS AND METHODS: A case-control study, for which we recruited 250 asthmatic adults and 250 age- and sex-matched controls. All subjects completed a questionnaire. Waist and hip circumference measurements, a lung function test and DNA genotyping were performed. In total, 50 incident cases and 50 matched controls who were non-smokers or had quit smoking for at least 1 year were selected in order to investigate SOD and CAT activity levels. RESULTS: In our study, we did not find a significant association between Mn-SOD Ala16Val, CAT C-262T and asthma. The level of SOD activity in new-onset asthma patients was significantly lower than in control subjects (p < 0.0005). The level of CAT activity in new-onset asthma patients was significantly higher than in control subjects (p < 0.0005). CONCLUSIONS: The levels of SOD and CAT activity were significantly related to adult asthma. SOD and CAT activity may be good tools to differentiate potential asthma sufferers. This would enable us to further investigate the mechanism of defective antioxidant enzymes in the context of asthma pathogenesis.

[Focal nodular hyperplasia of liver: a clinicopathologic study of 238 patients].

OBJECTIVE: To study the clinicopathologic features of focal nodular hyperplasia (FNH) of liver. METHODS: The clinical, radiologic, pathologic findings and follow-up data of 238 cases of FNH were retrospectively analyzed. RESULTS: The patients included 93 females and 145 males. The age of the patients ranged from 11 to 77 years (median = 39.1 years). Amongst the 233 patients who had clinical information available, 188 were asymptomatic, 216 had no history of hepatitis B and/or C infection and 232 had negative serum alpha-fetoprotein level. Amongst the 185 patients who had undergone radiologic examination, 123 (66.5%) were accurately diagnosed as such. Macroscopically, of the 284 lesions from 238 patients, the average diameter was 3.7 cm. Two hundred and fifteen cases (90.3%) were solitary, 172 cases were located in the right lobe and 115(40.5%) had central stellate fibrotic scars or lobulated cut surface. Histologically, 229 lesions belonged to classic type and 9 lesions were of non-classic type. The latter was further classified as the telangiectatic form (6 lesions) and the mixed hyperplastic and adenomatous form (3 lesions). There was no evidence of significant cytologic atypia. Follow-up data were available in 173 patients (72.7%). None of them died of the disease and 2 patients suffered from relapses after 2 and 4 years, respectively. CONCLUSIONS: FNH is a hyperplastic response of normal liver cells to local blood flow anomalies. It has no obvious sex predilection and more than 66% can be diagnosed accurately with radiologic examination. The lesions in the current study show no cytologic atypia.

[Study on clinicopathologic parameters of malignant behavior in gastrointestinal stromal tumors].

OBJECTIVE: To determinate the clinicopathologic parameters in predicting the malignant behavior of gastrointestinal stromal tumor (GIST). METHODS: Eight hundred and forty cases of GIST were retrospectively reviewed. The tumors were classified as malignant if they met any of the following criteria: evidence of gross dissemination (including liver metastasis and/or peritoneal spread), evidence of microscopic dissemination (including lymph node metastasis, infiltration to vessels, fat tissue, nerves and/or mucosal tissue), or disease relapse. The remaining cases were provisionally classified as tumors of uncertain biologic behavior. A number of morphologic parameters were then evaluated under light microscopy and univariate and multivariate analyses were adopted for this study. RESULTS: Histologic findings correlated with evidences of the following morphologic parameters were considered in accord with the criteria of the malignant behavior: mitotic count>or=10 per 50 high-power fields (P<0.01), muscle infiltration (P<0.01), coagulative necrosis (P<0.01), perivascular growth pattern (P=0.005) and remarkable nuclear atypia (P=0.014). Basing on the above criterion, 485 cases were re-classified as "malignant" and 355 cases "non-malignant". Follow-up data showed that the five-year disease-free survival and overall survival in the "non-malignant" group were 99.3% and 100% respectively, in contrast to 43.9% and 59.7% respectively in the "malignant" group (P<0.01). CONCLUSIONS: The set of clinicopathologic parameters is useful in predicting the malignant behavior of GIST and prognosis.

One-pot facile synthesis of CuNCs/RGO nanocomposite for the sensitive detection of heparin in human serum samples.

A simple and facile one-pot approach for the synthesis of copper nanoclusters decorated reduced graphene oxide (CuNCs/RGO) nanocomposite was proposed, in which the CuNCs attached to the surface of the reduced glutathione (GSH) functionalized RGO through ligand exchange via their thiol functionalities. The synthesized nanocomposite was verified by structural characterizations, and the further investigation of density functional theory (DFT) indicated that Cu3R2 cluster (R = C10H16O6N3S) with the lowest energy was the most stable structure in GSH-capped CuNCs. Although the CuNCs/RGO nanocomposite exhibited rather weak fluorescence, with the addition of heparin (Hep), the significant enhancement of fluorescence at 595 nm was achieved, which was developed to detect Hep in human serum samples with high selectivity and sensitivity. The mechanisms of fluorescence quenching of CuNCs/RGO nanocomposite and the sensing of Hep were discussed. The linear range was 0.1-10 µM with the detection limit of 26 nM in buffer solution containing 2% human serum sample, and satisfactory recovery in the range of 96.6%-104% was obtained, suggesting that the proposed method could applied to the detection of Hep in human serum samples.

The activity prediction of indole inhibitors against HCV NS5B polymerase.

Non-structural viral protein 5B (NS5B) is a viral protein in hepatitis C virus. Although various inhibitors against NS5B have been found, the activity prediction of similar untested inhibitors is still highly desirable. In this respect, the Tchebichef moments (TMs) calculated from the images of molecular structures were regarded as the independent variables while the inhibitory activity (pIC50 ) was the dependent variable, and the predictive model was established by means of stepwise regression. The R-squared of leave-one-out cross-validation (Q2 ) for the training set and the R-squared of prediction ( Rp2 ) for external independent test set were 0.919 and 0.927, respectively. The obtained model was also evaluated strictly. Compared with the multivariate curve resolution with alternating least squares (MCR-ALS) and the QSAR approaches derived from the literature, the proposed method is more accurate and reliable. This study not only provides an effective approach to predict the biological activity of RNA replication's inhibitors, but also extends the QSAR modeling technique.

Stage and histological grade of gastrointestinal stromal tumors based on a new approach are strongly associated with clinical behaviors.

Tumor stage and grade for gastrointestinal stromal tumors are poorly defined. To develop a better evaluation system, we assessed 12 clinical and pathological parameters in 613 patients with follow-up information. These parameters were classified into two gross spread parameters including liver metastasis and peritoneal dissemination, five microscopic spread parameters including lymph node metastasis, vascular, fat, nerve and mucosal infiltration, and five histological parameters including mitotic count > or =10 per 50 high-power fields, muscularis propria infiltration, coagulative necrosis, perivascular pattern and severe nuclear atypia. The 5-year disease-free survival and overall survival of 293 patients without any of these predictive parameters of malignancy were 99 and 100%, respectively. They were regarded as nonmalignant and further evaluations on the stage and grade of these tumors were not performed. At least one and at most seven predictive parameters of malignancy were identified in 320 patients. For these patients, the 5-year disease-free survival and overall survival rates were 44% (mean 6.7 years) and 60% (mean 9.3 years), respectively. The disease-free survival showed significant difference between patients with and without gross spread (P<0.0001), with and without microscopic spread (P=0.0009). Disease-free survival and overall survival were associated with the number of predictive parameters of malignancy in patients without gross spread (P<0.0001 for both disease-free survival and overall survival), but not in patients with gross spread (P=0.882 and 0.441, respectively). Malignant gastrointestinal stromal tumors could be divided into clinical stage I and II based on the absence and presence of gross spread, respectively. The degree of malignancy of patients in clinical stage I could be graded according to the number of predictive parameters of malignancy. Patients in clinical stage II were of the highest degree of malignancy regardless of the number of parameters. We found that the clinical stage and grade were strongly associated with prognosis.

[Study on the mechanism of imatinib-induced resistance in gastrointestinal stromal tumors].

OBJECTIVE: To investigate the mechanism of imatinib mesylate (IM) induced-resistance in the patients with gastrointestinal stromal tumors (GISTs) and treated with imatinib. METHODS: Eight patients with GIST treated with IM developed secondary IM resistance. A total of 16 tumor samples (pre-IM therapy) and 11 tumor samples (post-IM treatment) were available. Exon 9, 11, 13, and 17 of c-kit gene as well as exon 12 and exon 18 of PDGFRA gene were sequenced. RESULTS: In addition to the changes of baseline genotype, the IM-induced gene changes were concentrated in the kinase domain of c-kit gene in all 8 patients, 2 of them were located in the exon 13 of c-kit gene presenting with V654A, while 6 in exon 17 involving 816 and 820 to 823 codons. CONCLUSION: The mechanism of imatinib mesylate resistance after initial treatment with this agent in gastrointestinal stromal tumors is a novel mutation development in kinase domain of c-kit.

[Clinicopathologic analysis of pulmonary lymphangioleiomyomatosis].

OBJECTIVE: To improve the understanding and diagnosis of pulmonary lymphangioleiomyomatosis (LAM). METHODS: Fifteen cases of LAM of our hospital were presented and 73 cases reported in domestic literature from 1993 to 2008 were reviewed. By means of histological and immunohistochemical(IHC)studies, the clinical and pathological features of LAM were analyzed. RESULTS: All the 88 cases were female, with an average age of onset at (37 +/- 9) years. The main clinical manifestations included dyspnea (83/88, 94%), hemoptysis (48/88, 54%), pneumothorax (41/88, 47%), and chylothorax (28/88, 32%). High resolution computerized tomography (HRCT) showed thin-walled air-filled cysts throughout both lungs. Pathological features showed cystic changes in the lung, and abnormal smooth muscle cells (LAM cells) lined the airways, bronchioles, lymphatics and blood vessels leading to airflow obstruction and replacement of the lung parenchyma by cysts. In the autopsy case, extrapulmonary organs (eg, kidney, lymph nodes and soft tissues) were also involved. Abnormal manifestations in abdomen, including renal mass, retroperitoneal mass and retroperitoneal lymphadenopathy, were detected in 23 cases. CONCLUSIONS: LAM is a multisystem disease. Chest HRCT had confirmative value for diagnosis of LAM. In practice, chest HRCT, as well as other routine abdominal and pelvic imaging examinations, should be performed for child-bearing-age women with progressive dyspnea, hemoptysis, or spontaneous pneumothorax.

[Clinicopathological analysis of primary pulmonary cryptococcosis].

OBJECTIVE: To investigate the clinicopathological and CT features of primary pulmonary cryptococcosis (PC), and its underlying reasons for misdiagnosis. METHODS: The clinical data and pathological features of 52 cases of PC in Zhongshan Hospital from 1998 to 2008 were retrospectively reviewed. RESULTS: The 52 patients consisted of 36 males and 16 females aged from 17 to 80 years, with a median age of 48. The early symptoms were cough, phlegm and chest pain. Thirteen cases had a history of chronic diseases or malignant tumor or liver transplantation, including liver cancer and transplantation (n = 2), diabetes (n = 2), chronic hepatitis (n = 2), pituitary adenoma (n = 1), sarcoidosis (n = 1), diabetes concomitant with liver cancer and tuberculosis (n = 1), with tuberculous pleurisy (n = 1) and with hypertension (n = 1). Pulmonary nodules, either solitary or multiple, were the most common CT findings, present in 37 of the 45 cases. Cavitation was found in 4 cases, lobar consolidations in 3 cases, diffuse mixed pattern in 1 case. The CT diagnosis before surgery included malignant tumor (n = 27), pneumonia (n = 15) or tuberculosis (n = 3). Microscopically, cryptococcosis granuloma formation was found in 49 of the 52 cases, and the other 3 showed fibrosis with fungi. Cryptococcosis neoformans was detected in all the cases by mucicarmine and Grocott histopathological examination. CONCLUSIONS: Chest CT findings of PC in immunocompetent patients show a predominant pattern of localized and mixed lesions. PC does not have any specific clinical manifestations and image findings, and it is difficult to be differentiated from lung cancer, tuberculosis or pneumonia. The correct diagnosis relies on histopathological examination. CT guided percutaneous biopsy is useful in confirming the diagnosis.