Asymmetric Benzylic Allylic Alkylation Reaction of 3-Furfural Derivatives by Dearomatizative Dienamine Activation.

The dearomatizative dienamine-type ortho-quinodimethane species are smoothly generated between 2-alkyl-3-furfurals and chiral secondary amine catalysts, which undergo asymmetric benzylic allylic alkylation reactions with 2-nitroallylic acetates efficiently. A spectrum of densely functionalized 3-furfural derivatives are delivered in moderate to high yields with good to excellent diastereo- and enantioselectivity (up to 98 % yield, >19:1 d.r., >99 % ee). The latent transformations allow the facile production of some enantioenriched architectures, such as 1,1,2,2-tetraarylethanes and triarylmethanes, which are not easily available from other protocols.

[Purification Technology Optimization for Saponins from Ziziphi Spinosae Semen with Macroporous Adsorption Resin by Box-Behnken Design-Response Surface Methodology].

OBJECTIVE: To compare the purification effect of saponins from Ziziphi Spinosae Semen with different types of macroporous adsorption resin, and to optimize its purification technology. METHODS: The type of macroporous resins was optimized by static adsorption method. The optimum technological conditions of saponins from Ziziphi Spinosae Semen was screened by single factor test and Box-Behnken Design-Response Surface Methodology. RESULTS: AB-8 macroporous resin had better purification effect of total saponins than other resins, optimum technological parameters were as follows: column height-diameter ratio was 5: 1, the concentration of sample solution was 2. 52 mg/mL, resin adsorption quantity was 8. 915 mg/g, eluted by 3 BV water, flow rate of adsorption and elution was 2 BV/h, elution solvent was 75% ethanol, elution solvent volume was 5 BV. CONCLUSION: AB-8 macroporous resin has a good purification effect on jujuboside A. The optimized technology is stable and feasible.

Thrombin generation increasing with age and decreasing with use of heparin indicated by calibrated automated thrombogram conducted in Chinese.

OBJECTIVE: Calibrated Automated Thrombogram(CAT) is a test to monitor the generation of thrombin. It can be described by four parameters: lag time, peak thrombin, endogenous thrombin potential (ETP) and time to peak (ttPeak). This study aims to determine the normal ranges of CAT parameters in Chinese, and evaluate whether thrombin generation is correlated with the concentration of heparin/low molecular weight heparin. METHODS: Plasma from 120 healthy subjects were collected to determine the normal rangea of CAT parameters in Chinese. Normal plasma pool (NPP, n=25) spiked with different concentrations of heparin or enoxaparin were used to detecte CAT parameters. The overall and age specific normal ranges of CAT parameters were calculated using descriptive statistics method with mean±2SD. The correlation between CAT parameters and age or concentrations of heparin, enoxaparin were analyzed with linear regression model. RESULTS: The normal ranges for lag time, peak thrombin, ETP, ttPeak in the subjects were 3.648±2.465 min, 367.39±151.93 nmol/L, 2277±1030 nmol/L•min and 6.372±4.280 min respectively. Age was linearly correlated with lag time (r=-0.6583, P<0.0001), peak thrombin (r=0.4863, P<0.0001), ETP (r=0.3608, P<0.0014) and ttPeak (r=-0.6313, P<0.0001). The values of ETP/peak ratio were linearly correlated with concentrations of heparin. CONCLUSION: The normal ranges of four CAT parameters for Chinese were determined. CAT parameters are associated with age. ETP/peak ratio could be used to monitor the process of anticoagulation therapy.

[Genetic analysis and prenatal diagnosis for non-syndromic hearing impairment].

OBJECTIVE: To detect genetic mutations underlying non-syndromic hearing impairment (NSHI) and establish a method for prenatal diagnosis. METHODS: Sixty six NSHI patients were included in this study. DNA was extracted from peripheral blood. Genetic mutations were detected by gene chip analysis and direct sequencing of GJB2 gene. For 7 pregnant women at high risk, prenatal genetic diagnosis was provided. RESULTS: Fourteen cases (21.21%) were found to have GJB2 mutations by both methods (homozygous 235delC mutation in 3 cases, homozygous 176del16 mutation in 2 cases, 235delC and 299delAT compound heterozygous mutation in 2 cases, 299delAT and 176del16 compound heterozygous mutation in 1 case, c.339T > G and 313del12bp compound heterozygous mutation 1 case, and 235delC heterozygous mutation in 5 cases). 13 (19.70%) had SLC26A4 mutations (IVS7-2 A >G homozygous mutation in 2 cases, IVS7-2 A > G homozygous mutation in 2 cases, IVS7-2 A > G and 2168A > G compound heterozygous mutation in 3 cases, 2168A>G heterozygous mutation in 3 cases, and IVS7-2 heterozygous mutation in 3 cases); and 3 had mtDNA12S rRNA mutation (1555A > G mutation in 2 cases, 1494C > T mutation in 1 case). Prenatal diagnosis suggested that 3 fetuses have carried a heterozygous mutation. Two fetuses were detected as normal and confirmed to have normal hearing after birth. Two fetuses were found to have carried compound mutations of GJB2. CONCLUSION: Gene chip combined with GJB2 gene analysis is an accurate and effective method for the diagnosis of NSHI. The results can facilitate accurate prenatal diagnosis.

Remote Asymmetric Oxa-Diels-Alder Reaction of 5-Allylic Furfurals via Dearomatizative Tetraenamine Catalysis.

A previously unreported activation mode is developed through the generation of dearomatizative tetraenamine species between 5-allylic furfurals and a bifunctional amine-thiourea catalyst. The very remote ζ,η-alkenes perform as effective HOMO-raised dienophiles in inverse-electron-demand oxa-Diels-Alder cycloadditions with isatin-derived oxadiene substrates, delivering multifunctional spirocyclic oxindoles incorporating a dihydropyran skeleton in moderate to high yields with good to excellent enantio- and diastereoselectivity.

Detection of fetal epigenetic biomarkers through genome-wide DNA methylation study for non-invasive prenatal diagnosis.

The discovery of cell-free DNA fetal (cff DNA) in maternal plasma during pregnancy provides a novel perspective for the development of non­invasive prenatal diagnosis (NIPD). Against the background of maternal DNA, the use of the relatively low concentration of cff DNA is limited in NIPD. Therefore, in order to overcome the complication of the background of maternal DNA and expand the scope of cff DNA application in clinical practice, it is necessary to identify novel universal fetal­specific DNA markers. The GeneChip Human Promoter 1.0R Array set was used in the present study to analyze the methylation status of 12 placental tissue and maternal peripheral blood whole­genome DNA samples. In total, 5 fetus differential hypermethylation regions and 6 fetus differential hypomethylation regions were identified. In order to verify the 11 selected methylation regions and detect the differential CpG sites in these regions, a bisulfate direct sequencing strategy was used. In total, 87 fetal differential methylation CpG sites were identified from 123 CpG sites. The detection of fetal differential methylation DNA regions and CpG sites may be instrumental in the development of efficient NIPD and in the expansion of its application in other disorders.

[The role of Smad4 and MAPK proteins in signal transduction pathway in non-small cell lung cancer].

OBJECTIVE: To investigate the expression of Smad4 in non-small cell lung cancer (NSCLC), its correlation with MAPK (mitogen activated protein kinase) and their clinical significance in NSCLC. METHODS: Western blotting and RT-PCR were employed to test 42 resected lung cancers and normal lung tissues for the expression of Smad4. Imunohistochemistry was used to detect Smad4 and subtribes of MAPK in 71 paraffin samples. RESULTS: The level of protein and mRNA expression of Smad4 in lung cancer tissues were 0.2092 +/- 0.1308 and 0.3986 +/- 0. 1982, respectively, lower than those in normal tissues (0.7852 +/- 0.4386 and 1.1206 +/- 0.6772, P < 0.05). The expression of p38, ERK1 and Smad4 was associated with TNM staging (P = 0.000, 0.000 and 0.005, respectively) and JNK1 with tumor location (P = 0.028) and staging (P = 0.000). There was a correlation between p38 and Smad4 (P = 0.000). The expression of Smad4 (P = 0.0001), p38 (P = 0.0000) and JNK1 (P = 0.0208), tumor differentiation (P = 0.0059) and staging (P = 0.0000) were significantly correlated with prognosis of NSCLC by univariate analysis. Smad4 (P = 0.019), p38 (P = 0.044), tumor differentiation (P = 0.003), and staging (P = 0.020) were correlated with prognosis tested by multivariable analysis. Taking p38 and Smad4 together, we found that the negative expression of p38 and positive expression of Smad4 were associated with a better prognosis of NSCLC (P = 0.000). CONCLUSION: Smad4 could be of importance for the initiation and development of NSCLC. There is a significant correlation between main proteins of TGF-beta/smad4 and those of ras-MAPK signal transduction pathways. The expression of Smad4 is inhibited by p38. Smad4, as well as p38, tumor differentiation and staging can be used as prognostic factors of NSCLC.

[Expression of promyelocytic leukaemia protein in lung carcinomas and clinical significance thereof].

OBJECTIVE: To investigate the expression of promyelocytic leukaemia (PML) protein in lung carcinomas and the clinical significance thereof. METHODS: A tumor tissue microarray with lung samples from 148 patients with lung carcinoma and 5 patients with pulmonary benign tumor, and 7 patients with other benign diseases resected during operation. The cases with at least triplicate 0.6-mm cores per tumor or tissue were analyzed. Immunohistochemistry (SP method) was used to detect the expression of PML protein. The association between PML expression and post-operational survival was evaluated. RESULTS: Four cases of lung carcinoma were excluded because their available cores were less than 3. The remaining 144 lung carcinoma cases included 45 cases with squamous cell carcinoma, 62 with adenocarcinoma, 23 with small cell lung carcinoma (SCLC), 7 with large cell carcinoma, 5 with pleomorphic carcinoma, 1 with carcinoid, and 1 with adenosquamous carcinoma. Forty of the 144 cases were PML protein positive in the nucleus, 26 were PML protein positive in the cytoplasm, and 12 were PML protein positive in both the nucleus and cytoplasm. Among the 5 cases with benign lung tumors, two cases with pulmonary leiomyomas had strong expression of PML. Among the 7 normal pulmonary tissues, one case with bronchial mucosa had a moderate expression of PML on the nuclei in the basilar part, and the 6 cases with normal alveolar epithelium did not express PML. The rates of PML expression on cell nuclei were 31.4% and 8.7% respectively in the non-small cell lung carcinoma (NSCLC) and SCLC samples (chi(2) = 4.968, P = 0.026). Farish, but not strong, expression on cytoplasm was found in 39.1% cases of SCLC and in 14% cases of NSCLC (chi(2) = 6.609, P = 0.010). PML protein was found in 9 patients with SCLC, but absent in the other 14 cases. The 5-year cumulative survival rate of the PML positive SCLC patients was 50%, significantly higher than that of the PML-negative SCLC patients (23%, chi(2) = 3.931, P = 0.047). Lack of PML protein (RR = 0.127, 95% CI = [0.027, 0.592]) and senior pathologic TNM stage (RR = 3.148, 95% CI = [1.029, 9.631]) were two hazardous factors that influenced long-term survival of postoperative SCLC patients. CONCLUSION: As an important suppressor of tumor, PML may be a useful indicator to predict postoperative prognosis in patients with SCLC.

[Apoptosis induced by cantharidin in human pulmonary carcinoma cells A549 and its molecular mechanisms].

OBJECTIVE: To investigate the apoptosis-inducing effect of cantharidin in human lung cancer cells A549 and its molecular mechanisms. METHODS: MTT assay was used to determine A549 cells proliferation. Light and electron microscopy, FACScan, Annexin V-FITC staining and DNA gel electrophoresis were used to detect apoptosis. The expression of bcl-2, Bax and survivin were examined by Western blot. RESULTS: Cantharidin inhibited the proliferation of A549 cells. The cells treated with cantharidin showed a typical apoptotic morphology and hypodiploid peak before G(1) phase. Flow cytometry analysis with annexin quantitatively further confirmed the increase of cell apoptosis. DNA of treated A549 cells depicted a ladder pattern characteristic of apoptosis, indicating the presence of DNA fragmentation. Western blot assay showed that cantharidin increased the level of Bax expression and inhibited the level of bcl-2 and survivin expression. CONCLUSION: Cantharidin can induce A549 cells apoptosis mainly via regulation of Bax, bcl-2 and survivin expression.

Macrospirocyclic oligomer based on triphenylamine and diphenylphosphine oxide as a bipolar host for efficient blue electrophosphorescent organic light-emitting diodes (OLEDs).

A novel bipolar oligomer (TPA-PO)3 was prepared as a host material for efficient blue phosphorescent organic light-emitting diodes (OLEDs). Through the C-9s of the fluorene units, three triphenylamine units attached to diphenylphosphine oxide are connected in series to form a macrocyclic structure. The solution-processed phosphorescent device based on FIrpic and (TPA-PO)3 achieved a maximum current efficiency of 19.4 cd A(-1) and a maximum luminance of 11,500 cd m(-2) with a relatively low efficiency roll-off.

Surgery for young patients with lung cancer.

OBJECTIVES: To investigate the relationship between the clinical features and prognosis in young patients with lung cancer who underwent resection. METHODS: Statistical analysis was employed on sex, age, symptoms, diagnosis, treatment and prognosis, in 92 young cases younger than 40 years old among 930 cases with primary lung cancer who underwent surgery from January 1978 to December 1996. RESULTS: There were 92 young patients with lung cancer, accounting for 9.89% of the total cases. They were 71 male and 21 female patients, with the ratio of 3.38:1. The histological types were 34 squamous cell carcinomas (37%), 30 adenocarcinomas (33%), 26 small cell carcinomas (28%), and two large cell carcinomas (2%). On TNM staging, there were 30 cases in stage I (32.6%), 30 in stage II (32.6%) and 32 in stage III (34.8%). Lobectomy was conducted in 54 patients (59%), pneumonectomy in 36 (39%) and wedge-shaped resection in two cases (2%). The rate of pneumonectomy in young patients was significantly higher than that of 18% in older patients (>40) with lung cancer (P<0.01). 57 patients (62%) received absolutely curative resection; 28 cases (30%), relatively curative resection; seven cases (8%), non-curative resection. The postoperative 5-year-survival was 46% (42/92), in comparison with 34% (288/838) in patients older than 40 receiving operation during the same period, with significant difference between the two groups (P<0.05). The 5-year-survivals in patients with squamous cell carcinoma, adenocarcinoma, small cell carcinoma and large cell carcinoma were 68% (23/34), 30% (9/30), 38% (10/26) and 0 (0/2), respectively. The survival in squamous cell cancer was markedly higher than in adenocarcinoma (P<0.01) and in small cell carcinoma (P<0.05). There was no significant difference between adenocarcinoma and small cell cancer. The 5-year-survivals in stage I, II and III were 63% (19/30), 53% (16/30) and 22% (7/32), respectively. There was no significant difference between stage I and II, while remarkable difference was found between stage I and III (P<0.01), and between stage II and III (P<0.05). The 5-year-survival in patients undergoing absolutely curative resection was 67% (38/57), and 14% (4/28) in patients with relatively curative resection, with significant difference (P<0.01). No patient survived longer than the 5th postoperative year in seven cases receiving non-curative resection. CONCLUSIONS: Young patients with lung cancer were more often seen in male than in female. Squamous cell carcinoma accounted for the most part, no statistical difference, however, compared with adenocarcinoma and small cell carcinoma. The pneumonectomy rate in young patients was remarkably higher than that in patients older than 40. The postoperative 5-year-survival in young patients was considerably higher than in patients older than 40 who underwent surgery during the same period. Favorable prognosis was seen in patients with squamous cell carcinoma and undergoing absolutely curative resection, while worse outcome in stage III cancer.

Research on frequency of application with modern Chinese herbal medicine.

OBJECTIVE: To study the relationship between 500 kinds of commonly used Chinese herbal medicine and the classification of their efficacies in Chinese Materia Medica in relation to the common diseases listed in Internal Medicine. METHODS: Database retrieval frequency of the quantitative statistical method was adopted. First, the 8 980 kinds of Chinese herbal medicine recorded in Chinese Materia Medica were used as the original search objects, and 4 493 kinds which were cited in more than five articles were picked out and then rechecked for further title citations. Second, as judged based on the Criterion, the numbers of articles which included the medicines in the line of standards were examined. As a result, 500 species of Chinese herbal medicine were singled out based on their retrieval frequency and were then used for compilation of the classification statistics according to their efficacy and the common diseases in Internal Medicine. RESULTS: From the classification of Chinese medicines, herbs with wide efficiency and a meek nature had higher frequencies, but those which were not appropriate as decoctions had relatively lower frequencies. However, according to the average frequency, the Chinese herbal medicine for nourishing qi and tonifying blood, at 36,346 times and 34,544 times, respectively, were the most commonly used. Analyzed from the frequency of application of the Chinese medicine in the treatment of common diseases, most of the top 10 kinds of Chinese herbal medicine with the highest frequencies generally coincided with the 500 selected medicines. In addition, the Chinese medicines with clear pharmacological efficiency were easily isolated and purified to be made into injections, although other forms are more commonly used. CONCLUSION: The results of the research objectively reflected the current applications of Chinese herbal medicine, and could be used as references in teaching, research, clinical applications, and in compiling and increasing the drugs in textbooks and Pharmacopoeia.

[Clinicopathologic features of lung pleomorphic (spindle/giant cell) carcinoma--a report of 17 cases].

BACKGROUND & OBJECTIVE: Lung pleomorphic (spindle/giant cell) carcinoma is a rare epithelial malignant tumor. This study was to investigate its clinicopathologic and prognostic characteristics. METHODS: Clinicopathologic records of 17 patients with lung pleomorphic (spindle/giant cell) carcinoma were reviewed and compared with those of the patients with other histopathologic types of lung cancer treated in the same period. All patients underwent surgical resection. Kaplan-Meier method was used for survival analysis. Cox proportional hazards model was used for prognostic analysis. RESULTS: The 17 patients consisted of 15 men and 2 women with median age of 58 (45-78)û 5 at stage I, 3 at stage II, and 9 at stage III by pathologic TNM staging. Of the 17 cases of lung pleomorphic (spindle/giant cell) carcinoma, 2 were lung exclusive spindle cell carcinoma, 5 were lung carcinoma with spindle cells (combined with one kind of epithelial components, such as squamous cell carcinoma in 3 cases, adenocarcinoma in 1 case, and large cell carcinoma in 1 case), 10 were lung carcinoma with giant cell carcinoma (combined with one kind of epithelial components in 5 cases, two kinds in another 5 cases). Four patients at stage I survived free of tumor for more than 5 years. The median survival time was significantly shorter in lung pleomorphic (spindle/giant cell) carcinoma patients than in lung squamous cell carcinoma patients (36 months vs. 61 months, P=0.027), and was also significantly longer in patients with carcinoma containing spindle cells (including spindle cell carcinoma) than in patients with carcinoma containing giant cells (64 months vs. 18 months,P=0.026). Lymph node metastasis and carcinoma containing giant cells were poor prognostic factors of lung pleomorphic (spindle/giant cell) carcinoma. CONCLUSION: Lung carcinoma containing giant cells has multiple cells components, and has worse prognosis than lung carcinoma containing spindle cells and spindle cells carcinoma do.