Mixed-Linkage Donor-Acceptor Covalent Organic Framework as a Turn-On Fluorescent Sensor for Aliphatic Amines.

Amine determination is crucial to our daily life, including the prevention of pollution, the treatment of certain disorders, and the evaluation of food quality. Herein, a mixed-linkage donor-acceptor covalent organic framework (named DSE-COF) was first constructed by the polymerization between 2,4-dihydroxybenzene-1,3,5-tricarbaldehyde (DTA) and 4,4'-(benzo[c][1,2,5]selenadiazole-4,7-diyl)dianiline (SEZ). DSE-COF displayed superior turn-on fluorescent responses to primary, secondary, and tertiary aliphatic amines, such as cadaverine, isopropylamine, sec-butylamine, cyclohexylamine, hexamethylenediamine, di-n-butylamine, and triethylamine in absolute acetonitrile than other organic species. Further experiments and theoretical calculations demonstrated that the combination of intramolecular charge transfer (ICT) and photoinduced electron transfer (PET) effects between the DSE-COF and aliphatic amines resulted in enhanced fluorescence. Credibly, DSE-COF can quantitatively detect cadaverine content in actual pork samples with satisfactory results. In addition, DSE-COF-based test papers could rapidly monitor cadaverine from real pork samples, manifesting the potential application of COFs in food quality inspection.

Chiral Ionic Covalent Organic Framework as an Enantioselective Fluorescent Sensor for Phenylalaninol Determination.

Phenylalaninol (PAL) is a significant chemical intermediate widely utilized in drug development and chiral synthesis, for instance, as a reactant for bicyclic lactams and oxazoloisoindolinones. Since the absolute stereochemical configuration significantly impacts biological action, it is crucial to evaluate the concentration and enantiomeric content of PAL in a quick and convenient manner. Herein, an effective PAL enantiomer recognition method was reported based on a chiral ionic covalent organic framework (COF) fluorescent sensor, which was fabricated via one-step postquaternization modification of an achiral COF by (1R, 2S, 5R)-2-isopropyl-5-methylcyclohexyl-carbonochloridate (L-MTE). The formed chiral L-TB-COF can be applied as a chiral fluorescent sensor to recognize the stereochemical configuration of PAL, which displayed a turn-on fluorescent response for R-PAL over that of S-PAL with an enantioselectivity factor of 16.96. Nonetheless, the single L-MTE molecule had no chiral recognition ability for PAL. Moreover, the ee value of PAL can be identified by L-TB-COF. Furthermore, density functional theory (DFT) calculations demonstrated that the chiral selectivity came from the stronger binding affinity between L-TB-COF and R-PAL in comparison to that with S-PAL. L-TB-COF is the first chiral ionic COF employed to identify chiral isomers by fluorescence. The current work expands the range of applications for ionic COFs and offers fresh suggestions for creating novel chiral fluorescent sensors.

Thiophene-Containing Covalent Organic Frameworks for Overall Photocatalytic H2 O2 Synthesis in Water and Seawater.

H2 O2 is a significant chemical widely utilized in the environmental and industrial fields, with growing global demand. Without sacrificial agents, simultaneous photocatalyzed H2 O2 synthesis through the oxygen reduction reaction (ORR) and water oxidation reaction (WOR) dual channels from seawater is green and sustainable but still challenging. Herein, two novel thiophene-containing covalent organic frameworks (TD-COF and TT-COF) were first constructed and served as catalysts for H2 O2 synthesis via indirect 2e- ORR and direct 2e- WOR channels. The photocatalytic H2 O2 production performance can be regulated by adjusting the N-heterocycle modules (pyridine and triazine) in COFs. Notably, with no sacrificial agents, just using air and water as raw materials, TD-COF exhibited high H2 O2 production yields of 4060 µmol h-1 g-1 and 3364 µmol h-1 g-1 in deionized water and natural seawater, respectively. Further computational mechanism studies revealed that the thiophene was the primary photoreduction unit for ORR, while the benzene ring (linked to the thiophene by the imine bond) was the central photooxidation unit for WOR. The current work exploits thiophene-containing COFs for overall photocatalytic H2 O2 synthesis via ORR and WOR dual channels and provides fresh insight into creating innovative catalysts for photocatalyzing H2 O2 synthesis.

Fluorescence/Colorimetry/Smartphone Triple-Mode Sensing of Dopamine by a COF-Based Peroxidase-Mimic Platform.

Simple and accurate monitoring of urinary dopamine (DA) concentration is significant, which is helpful for the assessment or exclusion of catecholamine-producing tumors, such as pheochromocytoma and paraganglioma. Herein, a fluorescence/colorimetry/smartphone triple-mode sensing platform for DA determination was constructed using copper ion (Cu2+)-modified hydrazone-linked covalent organic frameworks (Cu-BTA-COF). Cu-BTA-COF with 21.67 wt % copper content exhibited peroxidase-mimic activity. After adding H2O2 and 1,3-dihydroxynaphthalene, the Cu-BTA-COF platform can sensitively and selectively detect DA in three modes with consistent results. In fluorescence/colorimetry/smartphone modes, the linear ranges of DA were 1-10, 0.2-40, and 1-10 µM, with related detection limits of 7.2, 8.6, and 23 nM, respectively. Moreover, the Cu-BTA-COF platform can be explored for DA determination in human urine samples with satisfactory recoveries (97.6-100.4%) in all the three modes, suggesting the potential practical application of the Cu-BTA-COF platform for DA detection in urine.

Ratiometric Fluorescent pH Sensor Based on a Tunable Multivariate Covalent Organic Framework.

Ratiometric detection of pH is always significant in environmental regulation, medical diagnosis, synthetic chemistry, and beyond. The construction of practical ratiometric pH sensors with reusability is still challenging. Herein, by exploiting a multivariate strategy, we first synthesized and reported a series of novel three-component covalent organic frameworks (COF-COOHX, X = 33, 50, and 67) through Schiff base reaction between 2-hydroxybenzene-1,3,5-tricarbaldehyde (HTA), 4,4'-diamino-3,3'-biphenyldicarboxylic acid (DBA), and 5,5'-diamino-2,2'-bipyridine (BPY) at various molar ratios (X = [DBA]/([BPY] + [DBA]) × 100 = 33, 50, and 67). COF-COOHX (X = 33, 50, and 67) displayed ratiometric pH sensing performance in acidic conditions with selectivity and repeatability. By tuning the molar ratio of DBA and BPY, the fluorescent properties, linear pH responsive ranges, and pKa values of COF-COOHX (X = 33, 50, and 67) can be regulated. Meanwhile, the two-component COF-COOH0 and COF-COOH100 did not exhibit ratiometric pH detection ability. Moreover, the constructed three ratiometric sensors can be applied to detect pH in drug solutions and carbonated drinks with satisfactory results. This work sheds new light on the design and fabrication of innovative ratiometric fluorescent sensors using COFs.

Cooperative Capture of Uranyl Ions by a Carbonyl-Bearing Hierarchical-Porous Cu-Organic Framework.

To efficiently capture the toxic uranyl ions (UO2 2+ ), a new hierarchical micro-macroporous metal-organic framework was prepared under template-free conditions, featuring interconnected multi-nanocages bearing carbonyl groups derived from a semi-rigid ligand. The material exhibits an unusually high UO2 2+ sorption capacity of 562 mg g-1 , which occurs in an intriguing two-steps process, on the macropore-based crystal surface and in the inner nanocages. Notably, the latter is attributed to the cooperative interplay of the shrinkage of the host porous framework induced by uranyl accommodation and the free carbonyl coordination sites, as shown by both single-crystal X-ray diffraction and a red-shift of the infrared [O=UVI =O]2+ antisymmetric vibration band.

Early medical abortion with self-administered low-dose mifepristone in combination with misoprostol.

AIM: The aim of the present study was to investigate the safety and efficacy of low-dose mifepristone combined with self-administered misoprostol for termination of early pregnancy. METHODS: A total of 533 women seeking medical abortion in early pregnancy (≤49 days since the last menstrual period) were divided randomly into hospital- (H-Mis, 250) and self- (S-Mis, 283) administered misoprostol groups. Women in two groups took 100 mg of oral mifepristone in hospital followed by 200 µg of sublingual misoprostol 24 h later in hospital or home. The primary outcome parameter was complete abortion without surgical intervention. Secondary outcomes were uterine bleeding, return of regular menses, side effects and patient acceptability. RESULTS: High rates of complete abortion were observed for both the H-Mis group (243/250; 94.8%) and the S-Mis group (266/283; 94.0%). No significant differences in outcomes (complete abortion/failure rates) or side effects were observed between the two groups. General satisfaction rates were similar for the two groups (H-Mis, 231/250, 92.4%; S-Mis, 263/283, 92.9%; P > 0.05). Higher convenience of administration (H-Mis, 211/250, 84.4%; S-Mis, 270/283, 95.4%; P < 0.05) and privacy protection (H-Mis, 214/250, 85.6%; S-Mis, 267/283, 94.3%; P < 0.05) satisfaction rates were obtained for the S-Mis group than for the H-Mis group. CONCLUSION: Self-administered sublingual misoprostol is as safe and effective as hospital-administered misoprostol following low-dose mifepristone to terminate early pregnancy (≤49 days of amenorrhoea) with fewer side effects.

Effect of Lymph Node Metastasis on Change of Positive Thyroglobulin Antibody in Differentiated Thyroid Carcinoma after Initial Treatment.

Objective To investigate the influence of lymph node metastasis on the change of positive thyroglobulin antibody(TgAb)in differentiated thyroid carcinoma after initial treatment.Methods We retrospectively analyzed the clinical data of 98 differentiated thyroid carcinoma patients with positive TgAb(≥115 IU/ml)before radioiodine(RAI)therapy.All of whom underwent total or near total thyroidectomy,neck lymph node dissection,and subsequent RAI therapy.Patients were divided into negative group(n=83)and non-negative group(n=15)according to the disappearance of positive TgAb or not after a mean follow-up of 21.0 months.Analysis of variance,χ2 test,and Mann-Whitney rank-sum test were applied to compare the basic clinical features including number of metastatic lymph nodes,lymph node metastasis rate and node stage,and dose of RAI ablation.The receiver operating characteristic curves were employed to evaluate the predictive values of TgAb levels(negative or positive)and optimal cut-off points.Multivariate analyses were further performed to explore the independent indicators for persistent positive TgAb. Results Compared with the negative group,the proportions of N1a and N1b in the non-negative group were significantly higher,with no N0 in the non-negative group(Fisher's Exact Test,P=0.032).The median metastatic lymph node rate was also significantly higher in the non-negative group(Mann-Whitney U=-3.498,P=0.000).The cut-off value for metastatic lymph node rate to predicting disappearance of positive TgAb was 24%,and its sensitivity was 71.4%.The multivariate analysis showed that only lymph node stage(OR=3.183,P=0.038)was the independent indicator for persistent positive TgAb. Conclusions Lymph node stage was an independent indicator for the disappearance of positive TgAb.A metastatic lymph node rate of higher than 24% may be predictive for the disappearance of positive TgAb.

Feasibility and effectiveness of unintended pregnancy prevention with low-dose mifepristone combined with misoprostol before expected menstruation.

STUDY QUESTION: What is the efficacy of maintaining or restoring non-pregnant status with low-dose mifepristone combined with misoprostol administered before expected menstruation? SUMMARY ANSWER: Low-dose mifepristone and misoprostol administered at the time of expected menstruation was effective and safe in maintaining or restoring non-pregnant status, with no obvious menstrual disturbance. WHAT IS KNOWN ALREADY: Menstrual regulation involves the medical or mechanical stimulation of uterine sloughing in women with up to 2-3 weeks of menstrual delay. Low-dose mifepristone plus misoprostol is efficacious for termination of ultra-early pregnancy (≤ 35 days of amenorrhoea) with no obvious menstrual disturbance. STUDY DESIGN, SIZE, DURATION: A total of 678 women fulfilled all criteria and were recruited. Seventeen women dropped out after deciding to remain pregnant and 11 others were lost to follow-up. Thus, data from 650 women who completed the procedure were included in analyses. Participants were enrolled at any time during their menstrual cycle and administered medication 1 day before expected menstruation. The end of the study was defined on a per-patient basis as the date of completion of the post-treatment menstrual cycle. The primary outcome was the efficacy of abortion induction (for pregnant women) or menstrual regulation (for non-pregnant women). PARTICIPANTS, SETTING, METHODS: Women with regular menstrual cycles (25-35 days) were voluntarily recruited for this study between February 2012 and December 2014. Serum ß-hCG was measured before mifepristone intake. Mifepristone (50 mg) was administered orally 1 day before expected menstruation and 200 µg misoprostol was administered orally on the day of expected menstruation. Efficacy, disturbance in bleeding patterns in the treatment and post-treatment cycles, satisfaction with the treatment, and subsequent contraception preference were analysed. MAIN RESULTS AND THE ROLE OF CHANCE: Retrospective analysis of serum ß-hCG levels at admission indicated that 23.3% (158/678) of the women were pregnant. The success rate for pregnancy termination was 98.6% (136/138). Two women (1.5%, 2/138) had ongoing pregnancy that was subsequently terminated surgically. The overall bleeding induction rate within 7 days was 98.3% (639/650), with 100% (138/138) in pregnant participants and 97.9% (501/512) in non-pregnant participants. Most pregnant and non-pregnant participants experienced no significant menstrual disturbance during the treatment [96.3% (131/136) versus 97.6% (489/501)] or post-treatment [97.8% (133/136) versus 98.4% (493/501)] menstrual cycle. The general rate of satisfaction with the treatment was 96.7% (618/639). Generally, 36.0% (230/639) of participants preferred to use the regimen as a routine contraception method versus the 64.0% (409/639) who preferred to use it as a remedy for pregnancy prevention after unprotected sex (P < 0.001). LIMITATIONS, REASONS FOR CAUTION: Study participants were recruited from a single region; further studies should include participants from multiple centres in different cities and nations. Given the uncertain efficacy of regimen reuse, the assessment of efficacy was based solely on the first treatment administration. Studies with larger samples and long-term follow-up may provide more data on whether repeated use of this regimen hampers its efficacy. WIDER IMPLICATIONS OF THE FINDINGS: Menstrual regulation with low-dose mifepristone and misoprostol at expected menstruation can be efficacious and highly acceptable to maintain or restore non-pregnant status, which may have potential for routine contraception.

Efficient production of cordycepin by engineered Yarrowia lipolytica from agro-industrial residues.

Cordycepin, a nucleoside compound with a variety of biological activities, has been extensively applied in the nutraceutical and pharmaceutical industries. The advancement of microbial cell factories using agro-industrial residues provides a sustainable pathway for cordycepin biosynthesis. Herein, the cordycepin production was enhanced by the modification of glycolysis and pentose phosphate pathway in engineered Yarrowia lipolytica. Then, cordycepin production based on economical and renewable substrates (sugarcane molasses, waste spent yeast, and diammonium hydrogen phosphate) was analyzed. Furthermore, the effects of C/N molar ratio and initial pH on cordycepin production were evaluated. Results indicated that the maximum cordycepin productivity of 656.27 mg/L/d (72 h) and cordycepin titer was 2286.04 mg/L (120 h) by engineered Y. lipolytica in the optimized medium, respectively. The cordycepin productivity in the optimized medium was increased by 28.81% compared with the original medium. This research establishes a promising way for efficient cordycepin production from agro-industrial residues.

Study of Dietary Emodin on Immune Defense in Megalobrama amblycephala against Aeromonas hydrophila.

This experiment aimed to investigate the effects of emodin on the total bacterial count and immune response in various tissues of Wuchang bream infected with A. hydrophila. The experimental diets were made by supplementing emodin at 0, 30, 100, and 150 mg kg-1 to basal (control) diet, respectively, and fed to fish with an initial weight of 50.4 ± 2.35 g. All fish were divided into five experimental groups: uninfected fish fed with basal control diet (negative control, NC), infected fish fed with the diet supplemented with 0 (positive control group, PC), 30 (30), 100 (100), and 150 mg/kg (150) of emodin. The fish were reared for 14 days and sampled at different time points. The results showed that the total bacterial count in the kidney, blood, and liver tissues of Wuchang bream infected with A. hydrophila was significantly affected by the supplementation and feeding time of emodin. At the beginning of the experiment, the difference in total bacterial count among the groups was not significant. On day 1, the total bacterial count in all groups was significantly higher (p < 0.05) than that in the negative control group. On day 4, the total bacterial count in all the emodin groups was significantly reduced, and the best bactericidal effect was observed in the 100 mg kg-1 group. In addition, emodin had a significant effect on the immune response of Wuchang bream after infection with A. hydrophila (p < 0.05). Compared with the other groups, the respiratory burst activity, tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1) content, and white blood cell count (WBC) in the 100 and 150 mg kg-1 groups could be restored to normal levels in the shortest time (p < 0.05). Furthermore, this study also measured the complement alternative pathway activity (ACH50), plasma superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content of the fish. The results showed that supplying 100 mg kg-1 emodin to the diet could significantly (p < 0.05) increase the ACH50 activity of the fish. Compared with the positive control (PC) group, the addition of emodin to the diet can inhibit the decrease in SOD activity and the increase in MDA content in the plasma of infected Wuchang bream. In conclusion, supplying 100 mg kg-1 emodin to the diet can enhance the ability of Wuchang bream to resist A. hydrophila infection by reducing the total bacterial count in tissues, increasing the activity of related immune enzymes, and promoting the secretion of cytokines. This provides a theoretical basis for production practice.

High-level de novo biosynthesis of cordycepin by systems metabolic engineering in Yarrowia lipolytica.

Cordycepin is a nucleoside antibiotic with various biological activities, which has wide applications in the area of cosmetic and medicine industries. However, the current production of cordycepin is costly and time-consuming. To construct the promising cell factory for high-level cordycepin production, firstly, the design and construction of cordycepin biosynthetic pathway were performed in Yarrowia lipolytica. Secondly, the adaptivity between cordycepin biosynthetic pathway and Y. lipolytica was enhanced by enzyme fusion and integration site engineering. Then, the production of cordycepin was improved by the enhancement of adenosine supply. Furthermore, through modular engineering, the production of cordycepin was achieved at 3588.59 mg/L from glucose. Finally, 3249.58 mg/L cordycepin with a yield of 76.46 mg/g total sugar was produced by the engineered strain from the mixtures of glucose and molasses. This research is the first report on the de novo high-level production of cordycepin in the engineered Y. lipolytica.

CDK5 Regulatory Subunit-Associated Protein 1-Like 1 Gene Polymorphisms and Gestational Diabetes Mellitus Risk: A Trial Sequential Meta-Analysis of 13,306 Subjects.

Objectives: The CDK5 regulatory subunit-associated protein 1-like 1 (CDKAL1) contributes to islet ß-cell function and insulin secretion by inhibiting the activation of CDK5. The current studies on the relationship between CDKAL1 polymorphisms rs7756992 A>G and rs7754840 C>G and the risk of gestational diabetes mellitus (GDM) have drawn contradictory conclusions. Materials and Methods: A meta-analysis with a fixed- or random-effects model was conducted to estimate the correlation between studied CDKAL1 polymorphisms and GDM risk with the summary odds ratio (OR) and 95% confidence interval (CI). In addition, trial sequential analysis (TSA) and false-positive report probability (FPRP) analysis were performed to confirm the study findings. Results: A total of 13,306 subjects were included in the present study. Meta-analysis results showed that the variant heterozygous and homozygous genotypes of the two polymorphisms were associated with increased GDM risk in comparison with the wild-type AA genotype (AG vs. AA: OR = 1.23, 95% CI = 1.08, 1.41, p = 0.002; GG vs. AA: OR = 1.47, 95% CI = 1.05, 2.05, p = 0.024 for rs7756992; and CG vs. GG: OR = 1.36, 95% CI = 1.13, 1.65, p = 0.002; CC vs. GG: OR = 1.76, 95% CI = 1.37, 2.26, p < 0.001 for rs7754840). The TSA confirmed a significant association between rs7754840 and the susceptibility to GDM because the cumulative Z-curve crossed both the conventional cutoff value and the TSA boundaries under the heterozygote and homozygote models. Conclusions: This study supported the finding that rs7756992 and rs7754840 are associated with susceptibility to GDM. However, further functional studies are warranted to clarify the mechanism.

Iguratimod promotes transformation of mononuclear macrophages in elderly patients with rheumatoid arthritis by nuclear factor-κB pathway.

BACKGROUND: The role of macrophages in rheumatoid arthritis (RA) and its mechanism have attracted much attention in RA pathogenesis. Macrophages accumulate in the synoviums of RA, and the proportion of M1 type pro-inflammatory macrophages is higher than that of M2 type anti-inflammatory macrophages, leading to the secretion of inflammatory molecules and the aggravation of inflammatory reaction, which has made macrophages a potential target of RA drugs. Iguratimod is a kind of cyclo-oxygenase-2 inhibitor that affects macrophage polarity. It is speculated that its anti-inflammatory and anti-rheumatic effects may be related to the regulation of macrophage M1/M2 ratio. AIM: To investigate the effects of Iguratimod on the polarity of mononuclear macrophages in elderly patients with RA. METHODS: Elderly patients with RA and joint effusion were selected, including 10 men and 25 women, with an average age of 66.37 ± 4.42 years. Patients were treated with oral administration of 25 mg Iguratimod (Iremod, State Food and Drug Administration Approval No. H20110084) twice daily for 12 wk. Disease Activity Score 28 and Health Assessment Questionnaire score were collected according to the disease severity before and after treatment. Venous blood and joint effusion fluid were collected, mononuclear macrophages were extracted and expression of cell surface markers CD86, CD64, CD163, and CD206 was analyzed by flow cytometry. The concentration of inflammatory factors interleukin (IL)-6, IL-1ß, transforming growth factor-ß, and IL-4 in the joint effusion fluid was analyzed by enzyme-linked immunosorbent assay. Expression of mononuclear cells inhibitor of nuclear factor-κB (IκB) and phosphorylated IκB in peripheral blood was analyzed by western blotting. RESULTS: Disease Activity Score 28 score and Health Assessment Questionnaire score of patients treated with Iguratimod decreased significantly. The percentage of cell surface markers CD86 and CD64 decreased significantly, and the percentage of CD163 and CD206 increased significantly (P < 0.05). The inflammatory factors IL-6 and IL-1ß decreased significantly, and transforming growth factor-ß and IL-4 increased significantly. Western blot analysis showed that mononuclear cell inhibitor of nuclear factor-κB in peripheral blood was significantly increased after treatment, and its phosphorylation level was significantly decreased (P < 0.05). CONCLUSION: Iguratimod can promote the transformation of mononuclear macrophages from M1 to M2 in elderly patients with RA by inhibiting the nuclear factor-κB pathway, thus improving symptoms of RA.

Association between functional genetic variants in retinoid X receptor-α/γ and the risk of gestational diabetes mellitus in a southern Chinese population.

To clarify the effect of retinoid X receptor-α/γ (RXR-α/γ) genes functional genetic variants (RXR-α rs4842194 G>A, RXR-γ rs100537 A>G and rs2134095 T>C) on the risk of gestational diabetes mellitus (GDM), a case-control study with 573 GDM patients and 740 pregnant women with normal glucose tolerance was performed in Guangxi area of China. An odds ratio (OR) with its corresponding 95% confidence interval (CI) was used to assess the strengths of the association between genetic variation and GDM. After adjustment of age and pre-BMI, the logistic regression analysis showed that the rs2134095 was significantly associated with GDM risk (CC vs. TT/TC: adjusted OR = 0.71, 95% CI = 0.56-0.90) in all subjects, and this result remained highly significant after Bonferroni's correction for multiple testing (P=0.004). The stratified analysis showed that rs2134095 was significantly associated with the risk of GDM among age > 30 years (adjusted OR = 0.61, 95% CI = 0.39-0.97), BMI > 22 kg/m2 (adjusted OR = 0.46, 95% CI = 0.30-0.70), systolic blood pressure (SBP) > 120 mmHg (adjusted OR = 1.96, 95% CI = 1.14-3.36), glycosylated hemoglobin A1c (HbA1c) < 6.5% (adjusted OR = 1.41, 95% CI = 1.11-1.78), TG ≤ 1.7 mmol/l (adjusted OR = 2.57, 95% CI = 1.45-4.53), TC ≤ 5.18 mmol/l (adjusted OR = 1.58, 95% CI = 1.13-2.22), high-density lipoprotein cholesterol (HDL-c) ≤ 1.5 mmol/l (adjusted OR = 1.70, 95% CI = 1.16-2.49) and low-density lipoprotein cholesterol (LDL-c) > 3.12 mmol/l (adjusted OR = 1.47, 95% CI = 1.08-2.00) subjects, under the recessive genetic model. We also found that rs2134095 interacted with age (Pinteraction=0.039), pre-BMI (Pinteraction=0.040) and TG (Pinteraction=0.025) influencing individual's genetic susceptibility to GDM. The rs2134095 T>C is significantly associated with the risk of GDM by effect of a single locus and/or complex joint gene-gene and gene-environment interactions. Larger sample-size and different population studies are required to confirm the findings.

[Clinicopathological features and prognostic implications of EGFR/c-Met gene copy number and protein expression in non-small cell lung cancer].

OBJECTIVE: To investigate the gene copy number and protein expression of EGFR/c-Met, so as to explore the relationship between them and the clinicopathological features and prognosis in non-small cell lung cancer (NSCLC) patients. METHODS: The expression of EGFR/c-Met protein was detected by immunohistochemical staining in NSCLC tissues of 61 cases. The level of EGFR/c-Met gene copy number was detected by fluorescent RT-PCR. RESULTS: The positive rates of EGFR and c-Met protein expression were 77.5% and 57.4%, respectively. These rates in well/moderately differentiated NSCLC tissues were 61.5% and 38.5%, all significantly lower than the data in poorly differentiated tissues (P < 0.05). The relative EGFR and c-Met gene copy number was 0.22 ± 0.22 and 0.20 ± 0.21 in smokers, respectively, all significantly higher than that in non-smokers (P < 0.05). The gene copy number of EGFR and c-Met in adenocarcinoma was 0.24 ± 0.26 and 0.23 ± 0.25, respectively, all higher than the data in squamous cell carcinoma (P < 0.05). There was a significant correlation in regard to the EGFR and c-Met protein expression, EGFR and c-Met gene copy number, and the protein expression vs. the gene copy number only in EGFR (P < 0.05). But there was no significant correlation between the c-Met protein expression and gene copy number (P = 0.259). There was a statistical significance between the postoperational median survival times (MST) of low EGFR gene copy number (48 months) and the high EGFR gene copy number (36 months) patients (P = 0.039). Similarly, there was also a significant difference between the MST of the low and high c-Met gene copy number patients (44 and 31 months, P = 0.022). CONCLUSION: There is a correlation between the EGFR and c-Met protein expression and the differentiation of NSCLC. The relative gene copy number is correlated with pathologic types and smoking of NSCLC patients, and it can be used in the prediction of prognosis.

[Recombinant lentivirus-mediated gene transfer of NT4-p53(N15)-Ant inhibits the growth of hepatocellular carcinoma cells in vitro].

OBJECTIVE: To construct a recombinant lentivirus vector containing fusion gene NT4-p53(N15)-Ant and transfer it into HepG2 cancer cells for gene therapy. METHODS: The gene of p53(N15)-Ant was obtained by T-vector method. After restriction enzyme digestion, the interest gene of p53(N15)-Ant was inserted in pBV220/NT4 vector and fusion gene of NT4-p53(N15)-Ant was subcloned into the plasmid of lentivirus and cotransferred into HEK-293 cells with helper plasmid. The recombinant lentivirus was produced by homologous recombination of the above mentioned two plasmids in HEK-293 cells and its titer was measured by plaque-forming. The expression of LV. NT4-p53-Ant in transfected HepG2 cells was finally confirmed by reverse transcription polymerase chain reaction (RT-PCR) procedure. The effect of LV. NT4-p53(N15)-Ant on HepG2 cells was measured by a colorimetric 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The inhibition effect on HepG2 cells of LV. NT4-p53(N15)-Ant and its potential mechanism was detected by light microscopy, electron microscopy, MTT, LDH-release assay and annexin V-PI double staining. RESULTS: The gene of p53(N15)-Ant was confirmed by restriction enzyme digestion and DNA sequencing. High titer of the recombinant lentivirus was obtained by homologous recombination in HEK-293 cell lines (1 x 10(11) pfu/ml), and the expression of NT4-p53(N15)-Ant gene in HepG2 cells was confirmed by RT-PCR. The viability of HepG2 cells was decreased to 83.4%, 46.9% and 33.9%, at 24 h, 48 h and 72 h, respectively, after infection by LV. NT4-p53(N15)-Ant. Compared with the LV. EGFP control group, there were significant differences (P < 0.01). The LDH level in HepG2 cells infected by LV. NT4-p53(N15)-Ant at 48 h, 72 h and 96 h after infection was 682 IU/L, 815 IU/L and 979 IU/L, respectively, significantly increased than that in the LV. EGFP group (P < 0.01), indicating the cell membrane destruction. CONCLUSION: The recombinant lentivirus vector encoding gene NT4-p53(N15)-Ant is successfully constructed in this experiment by molecular cloning and recombination in vitro techniques, and the results suggested that this fusion gene has an anti-tumor effect, which provides the basis for further research on recombinant adenovirus for cancer gene therapy.

[The clinical application of bi-level positive airway pressure noninvasive ventilator for home mechanical ventilation via tracheostomy in patients with amyotrophic lateral sclerosis].

OBJECTIVE: To study the feasibility of the bi-level positive airway pressure (BiPAP) non-invasive ventilator used in home mechanical ventilation for long-term tracheostomy-mechanical ventilation (TMV) in patients with amyotrophic lateral sclerosis (ALS). METHODS: Sixteen patients (12 men and 4 women, mean age 59 years) with ALS were selected for this study at Respiratory Department of the Shougang Hospital, Peking University from January 2002 to March 2008. After the disease had been controlled by anti-infective therapy and comprehensive treatment, the patients received TMV, through the improved ("Xiang's" connection) non-invasive BiPAP ventilator connected with tracheotomy tube, and on-going home mechanical ventilation (HMV). The blood gas was evaluated during invasive ventilation and non-invasive ventilation before discharge. Family members of the patients were trained for the use of non-invasive ventilators. The use of ventilators and the patients' condition were regularly followed and the survival rate calculated. Statistical analysis was carried out by using one-way ANOVA. RESULTS: There was no statistical difference in the blood gas before the use of non-invasive ventilator, 2 h and 1 d after the use of non-invasive ventilator, and before discharge, PaCO2 [(36+/-10), (42+/-11), (41+/-10), (42+/-11) mm Hg (1 mm Hg=0.133 kPa)], PaO2 [(84+/-11), (81+/-12), (87+/-14), (86+/-12) mm Hg], SaO2 [(96.7+/-1.3)%, (96.5+/-0.8)%, (96.8+/-1.2)%, (96.5+/-1.0)%] respectively, (F=1.21, 0.59, 0.97, 0.41, respectively, all P>0.05). All patients had no complaint of uncomfortable use, no intolerance to ventilators, and no ventilator breakdown. Fifteen patients were alive at the end of the follow-up (July 31, 2008). The mean time of using non-invasive ventilator was 39 months (range 4 to 66 months). CONCLUSION: For ALS patients who need long-term ventilation support, the use of BiPAP non-invasive ventilators is a safe and effective alternative for invasive ventilators.

Accumulation of hypoxia-inducible factor-1 alpha protein and its role in the differentiation of myeloid leukemic cells induced by all-trans retinoic acid.

BACKGROUND: The clinical activities of all-trans retinoic acid in the treatment of acute promyelocytic leukemia, a unique subtype of acute myeloid leukemia, have triggered extensive studies aimed at defining the mechanisms by which this compound induces differentiation of leukemic cells. Recent studies show that hypoxia-inducible factor-1 alpha (HIF-1 alpha) contributes to the differentiation of acute myeloid leukemia cells via transcriptional activity-independent mechanisms. We investigated whether all-trans retinoic acid affects HIF-1 alpha protein and whether this has a role in all-trans retinoic acid-induced differentiation. DESIGN AND METHODS: The acute myeloid leukemia cell lines NB4 and U937 were treated with all-trans retinoic acid, and HIF-1 alpha/HIF-1 beta mRNA and proteins were measured respectively by real-time quantitative reverse transcriptase polymerase chain reaction and western blotting. To investigate the role of HIF-1 alpha in all-trans retinoic acid-induced differentiation, NB4 cells, U937 cells, U937 cells in which HIF-1 alpha was induced by the withdrawal of tetracycline and U937 cells with stable expression of specific short hairpin RNA against HIF-1 alpha, Runx1, C/EBP alpha and PU.1, were treated with all-trans retinoic acid and/or the hypoxiamimetic agent cobalt chloride (CoCl(2)). Cellular differentiation was evaluated by morphological criteria and myeloid differentiation antigens. RESULTS: all-trans retinoic acid rapidly increased endogenous and inducible expressed or CoCl(2)-stabilized HIF-1 alpha protein in leukemic cells under normoxia. Importantly, suppression of HIF-1 alpha expression by specific short hairpin RNA partially but significantly inhibited all-trans retinoic acid-induced differentiation of the U937 cell line. Reciprocally, the differentiation induced by all-trans retinoic acid was significantly enhanced by conditional HIF-1 alpha induction and HIF-1 alpha-stabilizing CoCl(2) treatment. Furthermore, knock-down of PU.1, Runx1 and C/EBP alpha, three transcriptional factors crucial for normal hematopoiesis, greatly inhibited the differentiation cooperation of all-trans retinoic acid and HIF-1 alpha induction. CONCLUSIONS: This work provides the first demonstration that HIF-1 alpha, a protein rapidly responsive to all-trans retinoic acid, plays a role in all-trans retinoic acid-induced differentiation of leukemic cells. These observations shed new light on the molecular mechanisms underlying all-trans retinoic acid-induced differentiation of acute myeloid leukemia cells.

Integrated approach to pain management for a patient with multiple bone metastases of uterine cervical cancer.

Background Pain management for multiple bone metastases is complex and often requires multidisciplinary treatment. We herein describe patient-centered multidisciplinary pain management for metastatic cancer. CASE PRESENTATION: A 61-year-old woman with multiple bone metastases of uterine cervical cancer developed intractable low back pain. After external beam radiotherapy failed, we performed lumbar spinal intralesional curettage, pedicle screw fixation, and nerve decompression. However, the neuralgia persisted. We then percutaneously injected epirubicin into the intervertebral foramina under computed tomography guidance for L5 dorsal root ganglion destruction. Osteoplasty was performed under C-arm X-ray guidance; however, the sacrum was mistaken for the ilium, and treatment was ineffective. We administered zoledronic acid and strontium-89. The last resort was outpatient implantation of an epidural bupivacaine-morphine infusion system. A visual analog scale (VAS) was used for pain evaluation. Lumbar spinal intralesional curettage and fixation, epirubicin-induced ganglion destruction, and administration of zoledronic acid and strontium-89 decreased her VAS pain score from 7-8 to 3-4. Radiotherapy and nerve decompression and release were ineffective, as was osteoplasty because of the location error. The epidural infusion system decreased the VAS score from 7-8 to 2-3 and was highly efficient. Conclusions Multidisciplinary integrated treatment for metastatic cancer can be effective.