Impact of drug-linker on method selection for analytical characterization and purification of antibody-drug conjugates.

In addition to traditional characterisation methods of hydrophobic interaction (HIC) and reverse phase (RP) chromatography, an anion exchange chromatography (AIEX) was developed to analyse and purify antibody drug conjugates (ADCs). Since different drug antibody ratio (DAR) species may impact biological activity, therapeutic index, PK parameters or even potential immunogenicity, homogenous ADC DAR demands have been significantly increasing. To accelerate linker designs, drug screening and ADC DAR purification for in vitro and in vivo studies, we built the analytical toolbox including HIC, RP, AIEX, icIEF, SEC, and MS for downstream ADC DAR purification using HIC and AIEX. The established analytical methods can quickly assess the quality of ADC DAR profiles and provide important information to select the proper ADC DAR purification method. Since drug-linker structures can significantly affect ADC physicochemical properties, and highly impact on selections of analytical methods, we applied both HIC and AIEX characterisation and purification platforms to achieve ADC DAR homogenous. Our experiments also implied that unlike HIC, AIEX could be used to separate DAR4 positional isomers.

Risk factors for portal vein system thrombosis after partial splenic embolisation in cirrhotic patients with hypersplenism.

AIM: To determine risk factors for portal venous system thrombosis (PVST) after partial splenic artery embolisation (PSAE) in cirrhotic patients with hypersplenism. MATERIALS AND METHODS: Between March 2014 and February 2022, 428 cirrhotic patients with hypersplenism underwent partial splenic artery embolisation and from these patients 208 were enrolled and 220 were excluded. Medical records of enrolled patients were collected. Computed tomography (CT) images were reviewed by two blinded, independent radiologists. Statistical analyses were performed by using SPSS. RESULTS: Progressive PVST was observed in 18.75% (39/208) of cirrhotic patients after PSAE. No significant differences in peripheral blood counts, liver function biomarkers, and renal function were observed between the patients with progressive PVST and the patients without progressive PVST. The imaging data showed significant differences in PVST, the diameters of the portal, splenic, and superior mesenteric veins between the progressive PVST group and non-progressive PVST group. Univariate and multivariate analysis demonstrated portal vein thrombosis, spleen infarction percentage, and the diameter of the splenic vein were independent risk factors for progressive PVST. Seventeen of 173 (9.83%) patients showed new PVST; the growth of PVST was observed in 62.86% (22/35) of the patients with pre-existing PVST. Spleen infarction percentage and the diameter of the splenic vein were independent risk factors for new PVST after PSAE. CONCLUSION: The present study demonstrated portal vein thrombosis, spleen infarction percentage, and the diameter of the splenic vein were independent risk factors for PVST after PSAE in cirrhotic patients with hypersplenism.

[Genetic variations in four geographical isolates of Gohieria fusca based on cytochrome b and internal transcribed spacer genes].

OBJECTIVE: To investigate the genetic diversity and genetic differentiation of different geographical isolates of Gohieria fusca. METHODS: G. fusca isolates were sampled from Wuhu (WH), Bengbu (BB) and Bozhou cities (BZ) of Anhui Province and Jiaxing City of Zhejiang Province (JX). Mitochondrial cytochrome b (Cytb) and ribosomal internal transcribed spacer (ITS) genes were amplified in WH, BB, BZ and JX isolates of G. fusca using PCR assay. The gene sequences were edited and aligned using the software Chromas 2 and DNASTAR 1.00, and the haplotype, haplotype diversity (Hd) and nucleotide polymorphism (Pi) of each isolate were calculated using the software DnaSP 5.10.00. The genetic differentiation among isolates (Fst) and gene flow value (Nm) were estimated using the software MEGA 10.2, and a phylogenetic tree was built. Tests of neutrality and analysis of molecular variance (AMOVA) were performed using the software Arlequin 3.1 and a haplotype network was built based on the Median-Joining network using the software Network 10.2. RESULTS: PCR assay showed that the sizes of the Cytb and ITS genes were 372 bp and 1 301 to 1 320 bp, respectively. All four isolates of G. fusca presented high genetic diversity based on mitochondrial Cytb and ITS genes (Hd = 0.804, Pi = 0.006 91). AMOVA showed genetic differentiation among geographical isolates of G. fusca (Fst = 0.202 40, P < 0.05), and the genetic variation was mainly caused by intra-population variations (79.76%). Gene flow analysis showed a high level of gene flow among G. fusca isolates (Nm > 1). Tests of neutrality based on Cytb gene measured a Tajima's D value of -1.796 31 (P < 0.05) and a Fu's FS value of -3.293 98 (P < 0.05) in WH isolate of G. fusca, indicating population expansion in WH isolate of G. fusca. Haplotype network analysis and phylogenetic analysis revealed no remarkable geographical distribution pattern among different geographical isolates of G. fusca. All four isolates of G. fusca presented high genetic diversity (Hd = 0.985, Pi = 0.011 97). AMOVA showed moderate level of genetic differentiation between four isolates (Fst = 0.104 62, P < 0.05). The tests of neutrality based on ITS genes measured a Tajima's D value of -6.088 20 and a Fu's FS value of -1.935 99 (both P > 0.05) in the whole isolate of G. fusca, indicating no obviously population expansion. CONCLUSIONS: The four geographical isolates of G. fusca have high genetic diversity and remarkable genetic differentiation. Since a high level of gene flow is detected among different geographical isolates of G. fusca, no obvious geographical distribution pattern of G. fusca is found.

[Roles and mechanisms of m6A modification regulating RP11-426A6.5 in laryngeal squamous cell carcinoma].

Objective: To investigate the roles of N6-methyladenosine (m6A) modification in regulating RP11-426A6.5 in the development of laryngeal squamous cell carcinoma (LSCC). Methods: The methylation and expression levels of lncRNAs were identified and important lncRNAs were screened utilizing long non-coding RNA (lncRNA) m6A methylation microarray. Cancer and para cancer tissue samples were taken from 48 LSCC patients hospitalized to the Department of Otolaryngology-Head and Neck Surgery of the Second Affiliated Hospital of Harbin Medical University between January and September 2017. Expression profiling microarray was performed in 3 of 48 LSCC samples, and methylated RNA immunoprecipitation-quantitative PCR (MeRIP-qPCR) and quantitative real-time fluorescent PCR (qRT-PCR) were performed in the remaining 45 LSCC samples to verify the m6A modification and expression levels of RP11-426A6.5. Correlations between RP11-426A6.5 and clinical factors were anlysed. Laryngeal cancer cell line with low expression of RP11-426A6.5 was created in vitro using RNA interference (RNAi) technology. The 5-Ethynyl-2'-deoxyuridine (EdU) cell proliferation experiment, wound healing experiment, and transwell invasion experiment were used respectively to measure the proliferation, migration, and invasion of LSCC cells. The effect of RP11-426A6.5 down-regulation on the growth of transplanted tumors in vivo was verified by nude mice tumorigenesis assay. The Cancer Genome Atlas (TCGA) database and sequence-based RNA adenosine methylation site predictor (SRAMP) website were used to predict the enzymes and corresponding methylation sites. MazF digestion was chosen to validate the binding sites. RNAi technology was used to observe the changes in cell function after interfering with the expression of the corresponding genes of the modified enzymes. MeRIP-qPCR was used to detect the level of RP11-426A6.5 m6A cell line treated with actinomycin D was used to observe the stability of RP11-426A6.5. Results: RP11-426A6.5 methylation and expression levels were significantly higher in LSCC tissues than those in paracancerous tissues (methylation levels: 23.828±4.975 vs 20.280±3.607; expression levels: 1.197±0.314 vs 1.015±0.170, all P values<0.05). RP11-426A6.5 expression levels were closely correlated with T stage (T1-2: 1.081±0.298 vs T3-4: 1.306±0.292, χ2=5.35, P<0.05). The postoperative survival of patients with high RP11-426A6.5 expressions was significantly lower than that of patients with low RP11-426A6.5 expression (P=0.046). Assays in vitro and in vivo showed that the downregulation of RP11-426A6.5 significantly decreased the proliferation, migration, and invasion abilities of LSCC cells and the growth of transplanted tumors. The binding of methyltransferase-like 3 (METTL3), an m6A-modified enzyme, to the corresponding methylation site of RP11-426A6.5 enhanced its stability and mediated its regulation of malignant behaviors of LSCC cells. Conclusions: RP11-426A6.5 can regulate the malignant behaviors of LSCC cells, which is mediated by the m6A modification process involving in the methyltransferase METTL3.

Refeeding activates neurons in the dorsomedial hypothalamus to inhibit food intake and promote positive valence.

OBJECTIVE: The regulation of food intake is a major research area in the study of obesity, which plays a key role in the development of metabolic syndrome. Gene targeting studies have clarified the roles of hypothalamic neurons in feeding behavior, but the deletion of a gene has a long-term effect on neurophysiology. Our understanding of short-term changes such as appetite under physiological conditions is therefore still limited. METHODS: Targeted recombination in active populations (TRAP) is a newly developed method for labeling active neurons by using tamoxifen-inducible Cre recombination controlled by the promoter of activity-regulated cytoskeleton-associated protein (Arc/Arg3.1), a member of immediate early genes. Transgenic mice for TRAP were fasted overnight, re-fed with normal diet, and injected with 4-hydroxytamoxifen 1 h after the refeeding to label the active neurons. The role of labeled neurons was examined by expressing excitatory or inhibitory designer receptors exclusively activated by designer drugs (DREADDs). The labeled neurons were extracted and RNA sequencing was performed to identify genes that are specifically expressed in these neurons. RESULTS: Fasting-refeeding activated and labeled neurons in the compact part of the dorsomedial hypothalamus (DMH) that project to the paraventricular hypothalamic nucleus. Chemogenetic activation of the labeled DMH neurons decreased food intake and developed place preference, an indicator of positive valence. Chemogenetic activation or inhibition of these neurons had no influence on the whole-body glucose metabolism. The labeled DMH neurons expressed prodynorphin (pdyn), gastrin-releasing peptide (GRP), cholecystokinin (CCK), and thyrotropin-releasing hormone receptor (Trhr) genes. CONCLUSIONS: We identified a novel cell type of DMH neurons that can inhibit food intake and promote feeding-induced positive valence. Our study provides insight into the role of DMH and its molecular mechanism in the regulation of appetite and emotion.

MiR-20b is implicated in preeclampsia progression via the regulation of myeloid cell leukemin-1.

Preeclampsia (PE) may induce gestational failure, threatening a significant number of pregnant women. Recently, microRNAs (miRNAs) have been reported to participate in PE progression, whereas the precise functions and potential mechanisms of miR-20b in placental trophoblast cells as well as in PE progression remain poorly understood. In the present study, real-time quantitative polymerase chain reaction (RT-qPCR) analysis was used to detect expressions of miR-20b and myeloid cell leukemin- 1(MCL-1) mRNA. Cell viability was investigated by cell counting kit-8(CCK-8) assays. Cell invasion and migration abilities were determined by Transwell assays. Western blot was performed to detect MCL-1 protein expressions. The interaction between miR-20b and MCL-1 was investigated by bioinformatics analysis and luciferase activity assay. The results of the study demonstrated that miR-20b was highly expressed in placental tissues of patients with PE. Moreover, miR-20b overexpression inhibited HTR8/ SVneo cell proliferation, invasion and migration. Furthermore, MCL-1 was targeted by miR-20b, and MCL-1 restoration could partially attenuate the effect of miR-20b on HTR8/SVneo cells. In conclusion, the results indicate that miR-20b may contribute to PE through inhibiting proliferation, invasion and migration of placental trophoblast cells by targeting MCL-1. Therefore, miR-20b may be used as a notable biomarker for the diagnosis, prevention, and treatment of PE. MiR-20b targeting MCL-1 deserves further investigation in order to explore their potential role in PE.

Ferromagnetic frustration in ternary nitride ZnFe3N.

As a new antiperovskite nitride, ZnFe3N was synthesized and characterized by almost completely substituting iron atoms at corner positions of γ'-Fe4N. The magnetic interactions of the system with the space group Pm3[combining macron]m are fully investigated. The critical behavior was investigated based on the measured magnetic data around the ferromagnetic phase transition temperature. In this work, the values of critical exponents (ß, γ and δ) were obtained systematically using the Kouvel-Fisher method in the critical region. The Widom scaling law (δ = 1 + γß-1) and the scaling equation (m = f±(h)) were used to reveal the reliability of these values. The values of the critical exponents (ß = 0.325, γ = 1.228, and δ = 4.778) are different from those predicted by the three-dimensional (3D) Heisenberg model and mean-field model, and are very close to those of the 3D-Ising model. Combined with ESR analysis, the spin clusters induced by changes in chemical bonds are considered to be the cause for the existence of an anisotropic short-range ordered state in this ferromagnetic system.

MiR-1294 acts as a tumor inhibitor in cervical cancer by regulating FLOT1 expression.

Recently, important regulatory mechanisms of microRNAs (miRNAs) have been widely reported in human cancers including cervical cancer. The purpose of this study is to preliminarily clarify the function of miR-1294 in cervical cancer. The expression of miR-1294 or FLOT1 was detected using RT-qPCR or Western blot analysis. MTT, Transwell and luciferase reporter assays were used to explore the functional mechanism of miR-1294. The results showed that miR-1294 expression was decreased in cervical cancer. Functionally, overexpression of miR-1294 restrained the viability and metastasis of cervical cancer cells. MiR-1294 can also block EMT and suppress ß-catenin expression in cervical cancer cells. Additionally, FLOT1 was confirmed to be a direct target of miR-1294. The knockdown of FLOT1 impeded the progression of cervical cancer. More importantly, miR-1294 inhibited the occurrence of cervical cancer by interacting with FLOT1. In conclusion, miR-1294 inhibits cell viability, migration and invasion by suppressing FLOT1 expression.

[Clinicopathological features of myeloid sarcoma and DLBCL in the breast: a comparative study].

Objective: To study the clinicopathological features, diagnosis and differential diagnosis of myeloid sarcoma of the breast. Methods: Ten cases of myeloid sarcoma (MS) and 19 cases of diffuse large B cell lymphoma (DLBCL) of the breast were selected from Peking University People's Hospital from February 2005 to September 2019. The cases were evaluated by microscopy and immunohistochemistry basing on WHO classification (2008 and 2017). Results: For the 10 cases of MS, the mean and median age was 33.8 and 31 years (range 23 to 47 years) respectively. All patients presented with breast masses; six presented with B symptoms (6/10); and LDH level was elevated in four patients. The largest tumor dimension was 1.0 to 5.3 cm (mean 2.7 cm). All 10 patients had history of acute myeloid leukemia (AML), and in one patient, the AML occurred after chemotherapy for hydatidiform mole. One case was classified as M0, four were M2, two were M4 and three were M5. For the AML, all patients received chemotherapy and nine were treated by allogeneic hematopoietic stem cell transplant (allo-HSCT) and the breast masses occurred4 months to 2 years post-transplant. Using Ann Arbor staging, five cases were stage Ⅰ, three were stage Ⅱ, and 2 were stage Ⅳ. The MS was found in the left breast (two cases); right breast (three cases) and both breasts (five cases). Lymphocyte in peripheral blood, B symptom and site of lesion had statistical significance between myeloid sarcoma and DLBCL(P<0.05). The tumor cells were primitive, expressing MPO, CD43, CD117, etc. All ten patients had follow-up information, and the median survival period was 14.4 months (range 1 to 50 months). Seven patients died. The prognosis of patients with MS was worse than DLBCL(P=0.002). Conclusions: The clinical history, pathologic morphology, immunophenotyping and molecular studies are very important for diagnosing MS tumors in the breast, and MS may occur after allo-HSCT for AML. Tumor resection, chemotherapy, radiotherapy and donor lymphocyte infusion are recommended for treatment. The prognosis is poor.

Identification and comparative profiling of gonadal microRNAs in the adult pigeon (Columba livia).

1. MicroRNAs are small noncoding RNA molecules that play crucial roles in gene expression. However, the comparative profiling of testicular and ovarian microRNAs in birds are rarely reported, particularly in pigeon.2. In this study, Illumina next-generation sequencing technology was used to sequence miRNA libraries of the gonads from six healthy adult utility pigeons. A total of 344 conserved known miRNAs and 32 novel putative miRNAs candidates were detected. Compared with those of ovaries, 130 differentially expressed (DE) miRNAs were identified in the testes. Among them, 70 miRNAs showed down-regulation in the ovaries, while another 60 miRNAs were up-regulated.3. Combining the results of the expression of target gene measurements and pathway enrichment analyses, it was revealed that some DEmiRNAs from the gonad samples involved in sexual differentiation and development (such as cli-miR-210-3p and cli-miR-214-3p) could down-regulate AR (androgen receptor). Cli-miR-181b-5p, cli-miR-9622-3p and cli-miR-145-5p were highly expressed in both the ovaries and testes, which could co-target HOXC9, and were related to regulation of primary metabolic processes. KEGG enrichment analysis showed that DEmiRNAs may play biological and sex-related roles in pigeon gonads.4. The expression profiles of testicular and ovarian miRNA in adult pigeon gonads are presented for the first time, and the findings may contribute to a better understanding of gonadal expression in poultry.

Effect of ZEB2 silencing on cisplatin resistance in gastric cancer.

OBJECTIVE: To investigate the effect of ZEB2 silencing on cisplatin resistance in gastric cancer. MATERIALS AND METHODS: The resulting cell line, SGC7901/DDP, was transfected with ZEB2 siRNA, non-specific siRNA, or vehicle control. The effectiveness of ZEB2 silencing was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot. MTT viability assay was used to determine the cisplatin-sensitivity of cells. Cell apoptosis was measured by flow cytometry. RESULTS: A significant decrease in ZEB2 in mRNA and protein level was seen in cells transfected with ZEB2 siRNA, compared to that in cells transfected with non-specific siRNA or vehicle. Transfection with ZEB2 siRNA in cisplatin-resistant SGC7901/DDP cells resulted in a significant decrease in cell viability in response to the cisplatin treatment, and cell viability decreased with increasing cisplatin concentrations. A higher apoptotic rate was also seen in cells transfected with ZEB2 siRNA under cisplatin treatment. CONCLUSIONS: ZEB2 silencing can effectively make gastric cells sensitive to cisplatin treatment in vitro.

Critical behavior in tetragonal antiperovskite GeNFe3 with a frustrated ferromagnetic state.

Tetragonal GeNFe3 has a second-order ferromagnetic (FM) to paramagnetic transition at 76 K. Our integrated investigations indicate that the ground FM state is frustrated and the tetragonal symmetry is retained below 550 K based on the results of variable temperature X-ray diffraction. Critical behavior was analyzed by a systematic bulk magnetization study. The estimated critical exponents by three different methods (modified Arrott plot, the Kouvel-Fisher method, and critical isotherm analysis) conformably suggest that long-range magnetic coupling described by mean-field (MF) theoretical model is dominant in GeNFe3. The experimental M-T-H data collapse into two independent branches according to the scaling equations m = f±(h) with the renormalized magnetization m = ε-ßM(H, ε) and the magnetic field h = Hε-(ß+γ). The exchange distance is estimated as J(r) ∼ r-4.8 on the basis of the ß and γ values, which lies between the long-range MF model (r-4.5) and the short-range 3D Heisenberg (3DH) model (r-5). Our results indicate that the competition between local magnetic moments of iron 3d electronic state and itinerant covalent interactions of N-Fe bonds should be responsible for critical behavior in this system.

[Image features and clinical significance of pneumoconiosis with large shadow].

OBJECTIVE: To investigate the image features, categories, and clinical significances of Pneumoconiosis Ш with large shadow. METHODS: The research is based on the analysis of image features of 168 pneumoconiosis Ш patients who are directly diagnosed by qualified medical professionals in our institution. These image features are large shadows bigger than 2 cm×1 cm which can be observed by high kilovolt chest radiographs, DR chest radiography and chest CT scan. RESULTS: Large shadows bigger than 2×1 cm show noticeable imaging characteristics, like distribution of superior and middle lung regions (95.2%) , irregular shapes (97.6%) , splayed or sausage like changes on both sides perpendicular with ribs (89.2%) , small shadows of pneumoconiosis (98.8%) , and convergence of large shadow to hilum or mediastinal (53.6%). CT scan is significantly better than DR radiography on detection rate of focal emphysema, focal cavities and swollen lymph node of mediastinal. CONCLUSION: Combining the image features of high kilovolt chest radiographs, DR chest radiographs and CT scan are the main methods to identify the shadows, and thus direct diagnose pneumoconiosis Ш; CT scan plays an important role in differential diagnosis.

Preparation and characterization of a possible topological insulator BiYO3: experiment versus theory.

The Bi-Y-O system has been investigated by X-ray powder diffraction, electron diffraction, UV-vis and IR experiments. A metastable cubic high temperature phase of BiYO3 with fluorite-type structure has been structurally characterized for the first time and shows a large band gap of ∼ 5.9 eV. A unified description for the numerous structural variants discovered in the Bi-Y-O system is established within the symmetry breaking approach. This rich structural phenomenon makes the Bi-Y-O system a promising candidate in the search for new topological insulators for applications. On this basis, a long standing controversy on the phase diagram of the Bi-Y-O system has been solved. Our DFT calculations predict a high pressure phase for BiYO3 with perovskite (ABO3) structure and ordering of Bi and Y on the A and B sites, respectively. However, our analysis of the nature of the low energy electronic structure shows that this phase is not a suitable candidate for a topological insulator.

Impact of Comprehensive Rehabilitation Therapy on Trunk Controlling Ability of Children with Cerebral Palsy.

OBJECTIVE: This study aims to investigate the impact of comprehensive treatment on the trunk controlling ability (TCA) of children with cerebral palsy (CPC). METHODS: Twenty children with spastic cerebral palsy had truncal stability parameters in the sitting posture and the symmetry parameters of left-right-forward-backward centre of pressure evaluated by the stationary stability assessment module of the PK 254 P balance training instrument. After a six-month trunk control training with physical therapy technology and balancing instrument, the above indicators were re-evaluated, and the evaluations before and after the treatment were analysed to assess the therapeutic effect. RESULTS: The difference in the truncal stability parameters before and after the treatment was statistically significant (p < 0.05); the difference in the symmetry parameters of forward-backward centre of pressure was significant (p < 0.05). CONCLUSIONS: The balancing instrument training, combined with physical therapy technology training, could effectively improve the TCA of CPC, and the balancing instrument could also provide the objective and quantitative efficacy assessment toward the TCA of CPC.

Complete mitochondrial genome of the Chinese Hwamei Garrulax canorus (Aves: Passeriformes): the first representative of the Leiothrichidae family with a duplicated control region.

The Chinese Hwamei Garrulax canorus, a member of the family Leiothrichidae, is commonly found in central and southern China, northern Indochina, and on Hainan Island. In this study, we sequenced the complete mitochondrial genome of G. canorus. The circular mitochondrial genome is 17,785 bp in length and includes 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and two ribosomal RNA genes. In addition, two copies of highly similar putative control regions were observed in the mitochondrial genome. As found in other vertebrates, most of the genes are coded on the H-strand, except for one protein-coding gene (nad6; NADH dehydrogenase subunit 6) and eight tRNA genes (tRNA(Gln), tRNA(Ala), tRNA(Asn), tRNA(Cys), tRNA(Tyr), tRNA(Ser(UCN)), tRNA(Pro), and tRNA(Glu)). All the protein-coding genes start with ATG, with the exception of cox1 (cytochrome oxidase subunit 1), which starts with GTG. All tRNA genes have the potential to fold into the typical clover-leaf structure. Conserved sequences in three domains were observed in the two putative control regions. These results provide basic information for future phylogenetic analyses among species of the order Passeriformes.

Patient delay in the diagnosis and treatment of tuberculosis in China: findings of case detection projects.

OBJECTIVE: 1) To assess patient delay among new smear-positive pulmonary tuberculosis (PTB) patients in accessing health services in seven FIDELIS (Fund for Innovative DOTS Expansion through Local Initiatives to Stop TB) projects from 2003 to 2008 in China; 2) to compare treatment delay by province; and 3) to assess factors associated with delay. METHOD: Records of new smear-positive PTB patients were reviewed. Data sources were the consultation book, laboratory register, patient record, treatment card and the PWLAHS (people with limited access to health services) evaluation form. Data were collected using a standard questionnaire, cross-checked by staff from the sites and by the International Union Against Tuberculosis and Lung Disease (The Union) and analysed by The Union. RESULTS: Of the 75 401 new smear-positive PTB patients included in the study, 63-89% were PWLAHS. The average gross domestic product of the project sites and at national level were respectively US$557 and US$998. The median patient delay was 93 days (range 68-128). Delays were longer among females, older patients, rural residents and PWLAHS. Delayed access to health services was significantly associated with a greater number of symptoms. CONCLUSION: Patient delay in accessing health care in China was lengthy; TB care and control needs to be improved.

Objectif : Evaluer le délai d'accès aux services des santé de nouveaux patients atteints de tuberculose pulmonaire (TBP) à frottis positif détectés au sein de sept projets FIDELIS (Fund for Innovative DOTS Expansion through Local Initiatives to Stop TB) exécutés en Chine entre 2003 et 2008 ; 2) comparer les retards du traitement par province ; et 3) évaluer les facteurs associés aux retards.Méthode : Les dossiers des patients TPB à frottis positif ont été revus. Les données émanaient du registre des consultations, de celui du laboratoire, des dossiers des patients, des cartes de traitement et des formulaires d'évaluation des personnes ayant un accès limité aux services de santé (PWLAHS). Les données ont été recueillies grâce à un questionnaire standard et vérifiées par le personnel des sites et de l'Union Internationale contre la Tuberculose et les Maladies Respiratoires (L'Union) et analysées par L'Union.Résultats : De 75 401 nouveaux patients atteints de TBP à frottis positif, 63­89% étaient des PWLAHS. Le produit intérieur brut moyen des sites du projet et du niveau national était de 557 et 998 $US respectivement. Le délai médian d'accès aux soins était de 93 jours (extrêmes 68­128). Ce délai était plus long pour les femmes, les patients âgés, les résidents ruraux et ceux estimés être des PWLAHS. Ce délai d'accès aux services était significativement associé à un nombre de symptômes plus élevé.Conclusions : Le délai d'accès aux services de santé en Chine est long et offre des opportunités d'amélioration en matière de lutte contre la TB et de traitement.

Objetivo: 1) Evaluar el retraso en el acceso a los servicios de salud de los pacientes en quienes se diagnosticó tuberculosis pulmonar (TBP) con baciloscopia positiva en siete centros del Proyecto FIDELIS (Fondo para la Expansión Innovadora de DOTS por conducto de Iniciativas Locales a fin de poner Alto a la Tuberculosis) del 2003 al 2008 en la China; 2) comparar el retraso en el tratamiento entre provincias; y 3) evaluar los factores asociados con estos retrasos.Método: Se analizaron las historias clínicas de los pacientes con TBP y baciloscopia positiva. Las fuentes de los datos fueron los libros de consulta, el registro de laboratorio, las historias clínicas, las tarjetas de tratamiento y el formulario de evaluación de las personas con acceso limitado a los servicios de salud. Se recogieron los datos mediante un cuestionario normalizado y verificado que analizaron profesionales de los centros locales y de la Unión contra la Tuberculosis y las Enfermedades Respiratorias.Resultados: Se incluyeron en el estudio 75 401 casos nuevos de TBP con baciloscopia positiva. El producto interno bruto promedio por habitante a escala de los centros del proyecto era 557 USD y a escala nacional era 998 USD. De los pacientes con TB, entre el 63% y el 89% contaban con un acceso limitado a los servicios de salud. La mediana del retraso de los pacientes en acceder a la atención fue 93 días (extremos 68 a 128 días). Los retrasos fueron más prolongados en las mujeres, los pacientes de mayor edad, los residentes en zonas rurales y las personas que se calificaron con un acceso limitado a los servicios sanitarios. El acceso tardío a los servicios de salud se asoció de manera significativa con una mayor cantidad de síntomas.Conclusión: Se observó un retraso considerable de los pacientes para acceder a la atención de salud; esta situación ofrece oportunidades para mejorar la atención y el control de la TB.

Sorafenib combined with percutaneous radiofrequency ablation for the treatment of medium-sized hepatocellular carcinoma.

OBJECTIVE: Sorafenib, an oral multikinase inhibitor, is the proved therapy method for patients with advanced hepatocellular carcinoma (HCC). Based on heat delivery, Radiofrequency ablation (RFA) has been found to achieve complete neoplasm necrosis. It is the most widely performed percutaneous therapy for HCC. However, Study associated combined Sorafenib with RFA therapy for patients with advanced HCC has never been reported. The aim of present study is to explore the efficacy and safety of sorafenib combined with RFA therapy for the patients with medium-sized HCC. PATIENTS AND METHODS: A total of 62 patients diagnosed as HCC were involved in this study. All patients were randomly assigned to sorafenib and RFA (n=30) or RFA-alone (n=32) treatment groups. Treatment outcomes, including recurrence rates, time to progression (TTP) and adverse reactions induced by sorafenib were observed and recorded to assess the efficacy and safety of the combination method. RESULTS: During the overall follow-up period, the recurrence rate of the combination subgroup was 56.7% (17/30), and that of the RFA-alone subgroup was 87.5% (28/32) (p < 0.01). The median TTP was 17.0 months in the combination therapy vs. 6.1 months in the RFA-alone (p < 0.05). Hand-foot skin reactions were reported by 83.3% (25/30) of patients and 46.7% (14/30) reported diarrhea while the most adverse events (AEs) were mild to moderate in the combination subgroup. CONCLUSIONS: Sorafenib combined with RFA significantly decreased recurrence rates and prolonged the survival time of medium-sized HCC patients. The combination therapy is safer and more effective than the control without unexpected side effects. Furthermore, the earlier application, the better results were.

UPLC-MS/MS determination of phentolamine in human plasma and its application to a pharmacokinetic study.

A sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine phentolamine in human plasma. Sample preparation was accomplished through a simple liquid-liquid extraction with ethyl acetate. Chromatographic separation was carried out on an Acquity UPLC BEH C18 column using an isocratic mobile phase system composed of acetonitrile and 1% formic acid in water (33:67, v/v) at a flow rate of 0.45 mL/min. Mass spectrometric analysis was performed using a QTrap5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the positive ion mode. The MRM transitions of m/z 282.1 → 212.0 and m/z 237.1 → 194.2 were used to quantify for phentolamine and carbamazepine (internal standard, IS), respectively. The linearity of this method was found to be within the concentration range of 0.5-100.0 ng/mL with a lower limit of quantification of 0.5 ng/mL. Only 1.0 min was needed for an analytical run. This fully validated method was successfully applied to the pharmacokinetic study after oral administration of 60 mg phentolamine to 20 Chinese healthy male volunteers.