Preparation and optimization of dummy molecularly imprinted polymer-based solid-phase extraction system for selective enrichment of p-toluene sulfonate esters genotoxic impurities.

Sulfonate esters, one class of genotoxic impurities (GTIs), have gained significant attention in recent years due to their potential to cause genetic mutations and cancer. In the current study, we employed the dummy template molecular imprinting technology with a dummy template molecule replacing the target molecule to establish a pretreatment method for samples containing p-toluene sulfonate esters. Through computer simulation and ultraviolet-visible spectroscopy analysis, the optimal functional monomer acrylamide and polymerization solvent chloroform were selected. Subsequently, a dummy template molecularly imprinted polymer (DMIP) was prepared by the precipitation polymerization method, and the polymer was characterized in morphology, particle size, and composition. The results of the adsorption and enrichment study demonstrated that the DMIP has high adsorption capability (Q = 7.88 mg/g) and favorable imprinting effects (IF = 1.37); Further, it could simultaneously adsorb three p-toluene sulfonate esters. The optimal adsorption conditions were obtained by conditional optimization of solid-phase extraction (SPE). A pH 7 solution was selected as the loading condition, the methanol/1 % phosphoric acid solution (20:80, v/v) was selected as the washing solution, and acetonitrile containing 10 % acetic acid in 6 mL was selected as the elution solvent. Finally, we determined methyl p-toluene sulfonate alkyl esters, ethyl p-toluene sulfonate alkyl esters, and isopropyl p-toluene sulfonate alkyl esters in tosufloxacin toluene sulfonate and capecitabine at the 10 ppm level (relative to 1 mg/mL active pharmaceutical ingredient (API) samples) by using DMIP-based SPE coupled with HPLC. This approach facilitated the selective enrichment of p-toluene sulfonate esters GTIs from complex API samples.

Factors Associated with Acute Pulmonary Embolism in Patients with Hypoxia After off-Pump Coronary Artery Bypass Grafting: A Case-Control Study.

Purpose: This study aims to explore the factors linked to the occurrence of acute pulmonary thromboembolism (PE) within a cohort of patients exhibiting hypoxic saturation (oxygen saturation levels falling below 93%), subsequent to undergoing off-pump coronary artery bypass grafting (OPCABG). Methods: A retrospective case-control study was conducted. A total of 296 patients met the inclusion and exclusion criteria, divided into PE group (100 cases) and non-PE group (196 cases) according to whether they had PE or not. The preoperative and postoperative information of patients were collected and statistically analyzed. Results: The results from a multivariate logistic regression analysis indicated the following factors were independently linked to PE following OPCABG: history of smoking (OR = 3.019, 95% CI, 1.437-6.634, P = 0.004), preoperative arterial oxygen partial pressure ≤78.9 mmHg (OR = 3.686, 95% CI, 1.708-8.220, P = 0.001), presence of postoperative lower extremity deep venous thrombosis (OR = 4.125, 95% CI, 1.886-9.310, P < 0.001), elevated postoperative D-dimer levels >6.76 mg/l (OR = 8.078, 95% CI, 3.749-18.217, P<0.001), postoperative NT-BNP levels (OR = 1.001, 95% CI: 1.000-1.001, P = 0.011), and elevated postoperative pulmonary arterial pressure >33.0 mmHg (OR = 10.743, 95% CI: 3.422-37.203, P < 0.001). The developed nomogram exhibited a high predictive accuracy with an area under the curve of 0.913 (95% CI: 0.878-0.948). Conclusion: When patients have a history of preoperative smoking, decreased preoperative arterial oxygen pressure, postoperative lower limb DVT, increased postoperative pulmonary artery pressure, and elevated postoperative D-Dimer and NT pro-BNP levels, it is recommended to take perioperative preventive measures, timely diagnostic evaluation, and if necessary, anticoagulant treatment. In addition, the results of this study may improve the diagnostic sensitivity of medical staff for postoperative PE in OPCABG, thereby increasing the detection rate and potentially reducing the need for excessive medical imaging procedures.

A network pharmacology approach to decipher the mechanism of total flavonoids from Dracocephalum Moldavica L. in the treatment of cardiovascular diseases.

AIM OF THE STUDY: Cardiovascular disease (CVD) seriously endangers human health and is characterized by high mortality and disability. The effectiveness of Dracocephalum moldavica L. in the treatment of CVD has been proven by clinical practice. However, the mechanism by which DML can treat CVD has not been systematically determined. MATERIALS AND METHODS: The active compounds in DML were screened by literature mining and pharmacokinetic analysis. Cytoscape software was used to construct the target-disease interaction network of DML in the treatment of CVD. Gene ontology and signalling pathway enrichment analyses were performed. The key target pathway network of DML compounds was constructed and verified by pharmacological experiments in vitro. A hydrogen glucose deprivation/reoxygenation model was established in H9c2 cells using hypoxia and glucose deprivation for 9 h combined with reoxygenation for 2 h. The model simulated myocardial ischaemic reperfusion injury to investigate the effects of total flavonoids of Cymbidium on cell viability, myocardial injury markers, oxidative stress levels, and reactive oxygen radical levels. Western blot analysis was used to examine NOX-4, Bcl-2/Bax, and PGC-1α protein expression. RESULTS: Twenty-seven active components were screened, and 59 potential drug targets for the treatment of CVD were obtained. Through the compound-target interaction network and the target-disease interaction network, the key targets and key signalling pathways, such as NOX-4, Bcl-2/Bax and PGC-1α, were obtained. TFDM significantly decreased LDH and MDA levels and the production of ROS and increased SOD activity levels in the context of OGD/R injury. Further studies indicated that NOX-4 and Bax protein levels and the p-P38 MAPK/P38 MAPK andp-Erk1/2/Erk1/2 ratios were suppressed by TFDM. The protein expression of Bcl-2 and PGC-1α was increased by TFDM. CONCLUSIONS: Our results showed that DML had multicomponent, multitarget and multichannel characteristics in the treatment of CVD. The mechanism may be associated with the following signalling pathways: 1) the NOX-4/ROS/p38 MAPK signalling pathway, which inhibits inflammation and reactive oxygen species (ROS) production, and 2) the Bcl-2/Bax and AMPK/SIRT1/PGC-1α signalling pathways, which inhibit apoptosis.

A network pharmacology approach to decipher the total flavonoid extract of Dracocephalum Moldavica L. in the treatment of cerebral ischemia- reperfusion injury.

BACKGROUND AND OBJECTIVE: Cerebral ischemia-reperfusion injury (CIRI) is a major injury that seriously endangers human health and is characterized by high mortality and high disability. The total flavonoid extract of Dracocephalum moldavica L.(TFDM) in the treatment of CIRI has been proved by clinical practice. But the mechanism for the treatment of CIRI by TFDM has not been systematically revealed. STUDY DESIGN AND METHODS: The active compounds contained in TFDM were screened by literature mining and pharmacokinetic parameters, and the targets related to CIRI were collected by searching Drugbank, Genecards and OMIM databases. Cytoscape software was used to construct the protein interaction network of TFDM for the prevention and treatment of CIRI. Geneontology and signal pathway enrichment were analyzed. The key target pathway network of TFDM compounds was constructed and verified by pharmacological experiment in vitro. RESULTS: 21 active components were screened, 158 potential drug targets for the prevention and treatment of CIRI were obtained, 53 main targets were further screened in the protein-protein interaction network, and 106 signal pathways, 76 biological processes, 26 cell components and 50 molecular functions were enriched (P<0.05). Through the compound-target-pathway network, the key compounds that play a role in the prevention and treatment of CIRI, such as acacetin, apigenin and other flavonoids, as well as the corresponding key targets and key signal pathways, such as AKT1, SRC and EGFR were obtained. TFDM significantly decreased LDH, MDA levels and increased the NO activity levels in CIRI. Further studies have shown that TFDM increases the number of SRC proteins, and TFDM also increases p-AKT/ AKT. Molecular docking results showed that acacetin-7-O (- 6''-acetyl) -glucopyranoside, acacetin7-O-ß-D-glucopyranoside, apigenin-7-O-ß-D-galactoside respectively had good affinity for SRC protein. Acacetin-7-O (- 6''-acetyl) -glucopyranoside,acacetin-7-O-ß-D-glucuronide, acacetin7-O-ß-D-glucopyranoside had good affinity for AKT1 protein, respectively. CONCLUSION: Our research showed that TFDM had the characteristics of multi-component, multi-target and multi-channel in the treatment of CIRI. The potential mechanism may be associated with the following signaling pathways:1) the signaling pathways of VEGF/SRC, which promote angiogenesis, 2) the signaling pathways of PI3K/AKT, which inhibit apoptosis, and 3) acacetin-7-O (- 6''-acetyl) -glucopyranoside is expected to be used as a candidate monomer component for natural drugs for further development.

Histone Demethylase PHF8 Is Required for the Development of the Zebrafish Inner Ear and Posterior Lateral Line.

Histone demethylase PHF8 is crucial for multiple developmental processes, and hence, the awareness of its function in developing auditory organs needs to be increased. Using in situ hybridization (ISH) labeling, the mRNA expression of PHF8 in the zebrafish lateral line system and otic vesicle was monitored. The knockdown of PHF8 by morpholino significantly disrupted the development of the posterior lateral line system, which impacted cell migration and decreased the number of lateral line neuromasts. The knockdown of PHF8 also resulted in severe malformation of the semicircular canal and otoliths in terms of size, quantity, and position during the inner ear development. The loss of function of PHF8 also induced a defective differentiation in sensory hair cells in both lateral line neuromasts and the inner ear. ISH analysis of embryos that lacked PHF8 showed alterations in the expression of many target genes of several signaling pathways concerning cell migration and deposition, including the Wnt and FGF pathways. In summary, the current findings established PHF8 as a novel epigenetic element in developing auditory organs, rendering it a potential candidate for hearing loss therapy.

A population-based study on the prevalence of gestational diabetes mellitus in association with temperature in Taiwan.

BACKGROUND: Previous studies showing seasonal clustering of gestational diabetes mellitus (GDM) were conducted in the temperate or frigid zones and mostly included pregnant Caucasian women. This study aims to investigate the association of ambient temperature with prevalence of GDM in Taiwan, a sub-tropic country. METHODS: This population-based cohort study comprised women (n = 371,131) who gave births between 2013 and 2014; of which, 43,538 (11.7%) were diagnosed with GDM. The mean daily temperature and difference in temperature within a day was calculated over a 35-day period prior to GDM diagnosis or the first day of the 27th gestational week (for non-GDM subjects). Multiple logistic regression models with generalized estimation equation were performed to estimate the adjusted odds ratio (aOR) and 95% confidence interval (CI) of GDM in association with temperature. RESULTS: After controlling for potential confounders, summer and fall were associated with higher risk of GDM diagnosis, with aOR [95% CI] of 1.05 [1.04-1.07] and 1.04 [1.02-1.06] in reference to winter. Additionally, an increase of 1 °C from 14 °C to 27 °C was associated with an aOR of 1.03 [1.02-1.03]. The aOR greatly increased to 1.54 [1.48-1.60] after 28 °C. An increase of 1 °C difference within a day was associated with a reduced aOR at 0.90 [0.87-0.92]. CONCLUSION: A higher prevalence of GDM was associated with a higher daily temperature, but with a smaller difference in temperature within a day.

[Significance of differential diagnosis for sinonasal mass by CT value].

OBJECTIVE: To explore whether CT value is useful in identifying different disease in tumors of rhinosinus parenchyma.
 Methods: The data of preoperation noncontrast CT in 277 patients were retrospectively reviewed. The final diagnosis and classification were based on the result of surgical histopathological examination. The CT value range for different classification was calculated and was compared. All patients were re-diagnosed according to CT value range combined with pathological results by the same doctor team. The diagnosis rates according to CT value range were compared.
 Results: The CT value was (25.3±3.5) Hu in nasal polyp, (7.9±3.5) Hu in serous cyst, (42.2±4.7) Hu in mucocele, (40.7±5.3) Hu in papilloma, (112.3±10.9) Hu in fungus ball, (41.7±4.8) Hu in hemangioma, (51.2±9.9) Hu in malignant melanoma, and (47.1±9.9) Hu in squamous carcinoma. The CT value in nasal polyp is significantly higher than that in serous cyst, which was significantly lower than that in mucocele, papilloma, fungus ball, hemangioma, malignant melanoma and squamous carcinoma (all P<0.05); the CT value in serous cyst was significantly lower than that in other classification diseases (all P<0.05); the CT value in fungus ball was significantly higher than that in other classification diseases (all P<0.05); there was no significant difference in CT value among mucocele, papilloma, hemangioma, malignant melanoma, squamous carcinoma (all Pï¹¥0.05). The diagnosis rate was elevated (from 71.1% to 92.4%) according to CT value range, with significant difference (χ2=42.150, P<0.01).
 Conclusion: CT value in nasal polyp, serous cyst, fungus ball is different from other diseases, and the 3 diseases can be distinguished only by CT value range; the CT value in mucocele, papilloma, hemangioma, malignant melanoma and squamous carcinoma is similar, and their differential diagnosis should combine with imaging data and other clinical characters. The diagnosis rates can be improved when the CT value range is taken into account.

Massively Parallel Sequencing of a Chinese Family with DFNA9 Identified a Novel Missense Mutation in the LCCL Domain of COCH.

DFNA9 is a late-onset, progressive, autosomal dominantly inherited sensorineural hearing loss with vestibular dysfunction, which is caused by mutations in the COCH (coagulation factor C homology) gene. In this study, we investigated a Chinese family segregating autosomal dominant nonsyndromic sensorineural hearing loss. We identified a missense mutation c.T275A p.V92D in the LCCL domain of COCH cosegregating with the disease and absent in 100 normal hearing controls. This mutation leads to substitution of the hydrophobic valine to an acidic amino acid aspartic acid. Our data enriched the mutation spectrum of DFNA9 and implied the importance for mutation screening of COCH in age related hearing loss with vestibular dysfunctions.

[Bacterial culture analysis for patients with chronic rhinosinusitis with or without polyps].

OBJECTIVE: To determine the distribution of bacteria in patients with chronic rhinosinusitis, and to compare the bacteriologic features in middle meatus specimens between patients with nasal polyps (CRSwNP) and patients without nasal polyps (CRSsNP). 
 METHODS: We retrospectively analyzed the positive rate and types of bacterial culture in middle meatus specimens from 40 controls, 65 patients with CRSwNP, and 72 patients of CRSsNP. The specimens from the middle meatus were obtained during endoscopic sinus surgery.
 RESULTS: The positive rates of bacteria for CRSwNP, CRSsNP and the controls 81.9%, 80.0% and 82.5%, respectively, with no significant difference among the 3 groups. The common aerobe bacteria found in the specimens was Coagulase-negative staphylococci, Staphylococcus aureus, Streptococcus and Corynebacterium. The common anaerobe was Fusobacterium. The positive rates for aerobic and anaerobic bacteria showed no significant differences among the 3 groups.
 CONCLUSION: The distribution of bacteria in middle meatus specimens is not significantly different among CRSwNP, CRSsNP and the controls. Therefore, bacterial infection may not play a key role in the pathogenesis of CRS patients with and without nasal polyos.

[Association of vitamin D receptor ApaI and TaqI gene polymorphisms with pigmented pretibial patches in T2DM patients].

OBJECTIVE: To explore the association of vitamin D receptor ApaI and TaqI gene polymorphisms with pigmented pretibial patches (PPPs) in type 2 diabetes mellitus (T2DM) patients. METHODS: A total of 233 patients with T2DM were enrolled. Patients were assigned to PPPs group (n=106) or non-PPPs (n=127) according to the presence of PPPs. Allelic and genotypic comparisons of VDR-TaqI and VDR-ApaI between the different groups were evaluated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The genotype and allele gene frequency were similar at ApaI in two groups (P>0.05). The PPPs group showed a higher frequency of TT allele at TaqI in the T2DM (95%) when compared to that of non-PPPs group (87%), while the frequency of Tt and tt allele were lower in PPPs group than non-PPPs group in T2DM (5% vs 13%, all P<0.05). The frequency of T allele were higher at TaqI in PPPs group (98%) than non-PPPs group (93%) in T2DM, while PPPs had a lower frequency of t allele (2% vs 7%, all P<0.05). CONCLUSION: It seems that gene polymorphisms of VDR-TaqI, not VDR-ApaI, contributes to the risk of PPPs in T2DM patients in the Han nationality in Tianjin area.

[Free radial forearm flap for reconstruction of head and neck soft tissue defects after tumor resection].

OBJECTIVE: To explore the clinical effect of the free radial forearm flap on repairing tissue defects and reconstructing functions after tumor resection.
 METHODS: From January, 2003 to December, 2011, 70 patients, including 43 squamous cell carcinomas of tongue, 12 buccal cancers, 5 carcinomas of the soft palate, 4 basal cell carcinomas of external nose, 3 lower lip cancers, 2 upper lip cancers, and 1 posterior wall of hypopharynx carcinoma, with the soft tissue defects in the head and neck underwent reconstructive operations with the free radial forearm flap after the malignant tumor resection. The area of defects ranged from 5 cm × 4 cm to 14 cm × 8 cm with the process of diseases from 4 to 30 months. The technique for grafting the free radial forearm flap and the appearance at sites of the donor and recipient, and the influence on the anatomy and function in both local sites were analyzed.
 RESULTS: In the 70 patients, only 1 case of flap appeared necrosis due to venous reflux obstacle, and the remaining (98.4 ﹪) survived. During the follow-up for 12-36 months, one case of hypopharyngeal carcinoma died from distant metastasis a year later, 2 cases of tongue cancer died of cardiovascular accident. Morphology and function for the sites at donor and recipient were satisfactory.
 CONCLUSION: Free radical forearm flap is a good choice for the repair and functional reconstruction for tissue defects after tumor resection.

[STAT signaling pathway mediates high glucose induced cardiac fibroblasts proliferation and collagen deposition in vitro].

OBJECTIVE: To observe the signal transducers and activator of transcriptions (STATs) protein expression changes and investigate the functional role of STATs pathway in case of high glucose-induced cardiac fibroblasts (CFs) proliferation and collagen deposition in vitro. METHODS: Rat cardiac fibroblasts were isolated from 1- to 3-day-old SD rats, cells from the second to fourth passages were used for the experiment. CFs were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 mmol/L glucose (NG), 5.5 mmol/L glucose plus 19.4 mmol/L mannose (OC) or 25 mmol/L glucose (HG) in the presence of absence of STAT1 inhibitor (fludarabine, FLU) and STAT3 inhibitor (S3I-201). After 24 h and 48 h culture in vitro, the proliferation of CFs was measured by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. After 12 h and 24 h culture in vitro, the production of type I and III collagen was evaluated using real-time quantitative PCR and ELISA. After 0, 30, 60 and 120 min culture in vitro, the phosphorylated expression of STAT1 and STAT3 was analyzed by Western blot. RESULTS: CFs proliferation was significantly enhanced post 24 h and 48 h HG stimulation, and procollagen I and III mRNA expression was significantly upregulated post 12 h and 24 h HG stimulation. Deposition of collagen I and III was also significantly increased post 24 h and 72 h HG stimulation. STAT1 phosphorylation in CFs was increased after 120 min HG stimulation and STAT3 phosphorylation in CFs was increased post 60 min and 120 min HG stimulation. FLU and S3I-201 could inhibit HG-induced CFs proliferation and suppress of which was stimulated by FLU and S3I-201 could both suppress upregulated procollagen I and III mRNA expression and the deposition of collagen types I and III post HG stimulation. STAT1 phosphorylation inhibition resulted in less mRNA downregulation of procollagen type III than that of procollagen type I post 12 h HG stimulation. The STAT3 phosphorylation inhibition resulted in more significantly upregulated procollagen type III mRNA expression than procollagen type I mRNA expression at 12 h post HG stimulation. CONCLUSION: HG could enhance the protein expression of phosphorylated STAT1 and STAT3 in CFs, which are responsible for HG-induced increased CFs proliferation and collagen deposition in vitro.

STAT1/3 and ERK1/2 synergistically regulate cardiac fibrosis induced by high glucose.

BACKGROUND: Hyperglycaemia promotes the proliferation of cardiac fibroblasts (CFs) and collagen synthesis in CFs. However, the molecular mechanism underlying the effects of HG on proliferation and collagen synthesis of CF, is not completely understood. OBJECTIVES: The objectives of the present study were to determine whether the STAT proteins has a functional role in high glucose-induced proliferation of CFs and collagen synthesis in vitro and whether the STAT signaling pathway and MAPK signaling pathway have synergetical effects on high glucose-mediated cardiac fibroblasts proliferation and collagen synthesis. METHODS: Rat CFs were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 or 25 mmol/L D-glucose, in the presence of absence of STAT1 inhibitor Fludarabine, STAT3 inhibitor S31-201 and ERK1/2 inhibitor PD98059. Proliferation were measured by the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, the production of Type I and III collagen was evaluated using real-time quantitative RT-PCR and ELISA, and the phosphorylation expression of STAT1 and STAT3 were analyzed by Western blot. RESULTS: High glucose treatment promoted the proliferation of cardiac fibroblasts and collagen types I and III synthesis. High glucose treatment induced STAT1 and STAT3 phosphorylation in cardiac fibroblasts, the mode and level of STAT1 and STAT3 phosphorylation were significantly different. Fludarabine and S31-201 could both inhibited high glucose stimulated proliferation of cardiac fibroblasts and collagen types I and III synthesis with different effects. Combination of Fludarabine and PD98059 or combination of S31-201 and PD98059 both exhibited stronger inhibitions on proliferation of cardiac fibroblasts and collagen types I synthesis, but the effects and functional modes are different. CONCLUSION: Both STAT1 and STAT3 mediate the proliferation of cardiac fibroblasts and collagen synthesis induced by high glucose. STAT1 and STAT3 both have synergetic effects with ERK1/2 on regulating proliferation of cardiac fibroblasts and collagen types I synthesis.

Free jejunal graft for reconstruction of defects in the hypopharynx and cervical esophagus following the cancer resections.

OBJECTIVES: The reconstruction of esophagus defects after hypopharyngeal and cervical esophageal carcinoma resection is an ongoing problem. The objective of this article was to investigate the techniques of the free jejunal graft for the reconstruction of hypopharyngeal and cervical esophagus and discuss the outcome related to the procedures. SUBJECTS AND METHODS: From July of 2005 to December 2007, seven patients with hypopharyngeal and cervical esophageal cancer underwent free jejunal graft reconstruction of the hypopharyngeal and cervical esophagus. Their clinical data were retrospectively analyzed. All patients received postoperative radiotherapy and were followed up for 7-24 months. RESULTS: Despite the multistep and time-consuming procedure, free jejunal graft survival was 100%. Operation-induced complications did not occur in six patients. One patient developed pharyngeal fistula. CONCLUSION: The present experience supports the use of free jejunal grafts in reconstruction of the hypopharyngeal and cervical esophagus defects after exenteration of the central compartment of the neck. A high successful rate with low incidence of complications in reconstruction of the hypopharyngeal and cervical esophagus was obtained in this study.

Soluble decoy receptor 3 induces angiogenesis by neutralization of TL1A, a cytokine belonging to tumor necrosis factor superfamily and exhibiting angiostatic action.

TL1A is a member of the tumor necrosis factor superfamily and plays an important role in regulating endothelial cell apoptosis. A previous study shows TL1A is able to interact with death receptor 3 and decoy receptor 3 (DcR3). Here, we demonstrate that DcR3 is able to induce angiogenesis in human umbilical vein endothelial cells (HUVECs). DcR3 promotes HUVEC proliferation and migration and up-regulates matrix metalloproteinase-2 mRNA expression and enzyme activity. Furthermore, DcR3 enhances EC differentiation into cord vascular-like structures in vitro, as well as neovascularization in vivo. The effects of DcR3 on HUVECs are also mimicked by anti-TL1A and antideath receptor 3 antibodies. In contrast, human aortic endothelial cells, which do not express TL1A, are not responsive to DcR3 treatment, including cell proliferation, migration, and angiogenic differentiation. These data demonstrate DcR3 might not only help tumor cells to escape immune surveillance but also induce angiogenesis by blocking TL1A action in endothelial cells. The pathological role of DcR3 in promoting cancer progress raises the possibility to target DcR3 for antiangiogenic therapy in the future.