Sleep deprivation drives brain-wide changes in cholinergic presynapse abundance in Drosophila melanogaster.

Sleep is an evolutionarily conserved state that supports brain functions, including synaptic plasticity, in species across the animal kingdom. Here, we examine the neuroanatomical and cell-type distribution of presynaptic scaling in the fly brain after sleep loss. We previously found that sleep loss drives accumulation of the active zone scaffolding protein Bruchpilot (BRP) within cholinergic Kenyon cells of the Drosophila melanogaster mushroom body (MB), but not in other classes of MB neurons. To test whether similar cell type-specific trends in plasticity occur broadly across the brain, we used a flp-based genetic reporter to label presynaptic BRP in cholinergic, dopaminergic, GABAergic, or glutamatergic neurons. We then collected whole-brain confocal image stacks of BRP intensity to systematically quantify BRP, a marker of presynapse abundance, across 37 neuropil regions of the central fly brain. Our results indicate that sleep loss, either by overnight (12-h) mechanical stimulation or chronic sleep disruption in insomniac mutants, broadly elevates cholinergic synapse abundance across the brain, while synapse abundance in neurons that produce other neurotransmitters undergoes weaker, if any, changes. Extending sleep deprivation to 24 h drives brain-wide upscaling in glutamatergic, but not other, synapses. Finally, overnight male-male social pairings induce increased BRP in excitatory synapses despite male-female pairings eliciting more waking activity, suggesting experience-specific plasticity. Within neurotransmitter class and waking context, BRP changes are similar across the 37 neuropil domains, indicating that similar synaptic scaling rules may apply across the brain during acute sleep loss and that sleep need may broadly alter excitatory-inhibitory balance in the central brain.

[Diagnostic value of high frequency oscillation in localization of type â ¡ focal cortical dysplasia epilepsy].

Retrospective analysis was conducted on 9 patients with type Ⅱ focal cortical dysplasia (FCD) who underwent stereo-electroencephalography (SEEG) implantation in the Department of Neurosurgery of the First Affiliated Hospital of Fujian Medical University from November 2020 to February 2023. The onset area, onset time, and frequency of high-frequency oscillations (HFO) were analyzed and the correlation of HFOs with interictal, preictal, and ictal periods. SEEG recordings of 80-500 Hz HFOs were observed in both interictal and ictal periods in 9 patients, with 6 patients exhibiting fast ripples (FR) in the range of 250-500 Hz. Surgical resection of the seizure onset area and FR-generating electrodes was performed, and postoperative follow-up for over 2 years indicated Engel I in 5 cases. 6 patients showed continuous discharge during the preictal period, and the distribution index of continuous discharge was positively correlated with seizure frequency. HFOs in the range of 80-500 Hz were present in all four seizure onset patterns during the ictal period. The onset area and FR-emitting electrode were surgically removed in 6 patients with continuous discharge and overlapping HFOs during the preictal period, with 5 cases of Engel I. Type Ⅱ FCD discharges exhibited complexity, high discharge indices, and a close association with HFOs. Compared with the spike wave, the electrode range of HF is more limited, and the incidence of HF before attack is significantly increased, which is closely correlated with the onset area. The simultaneous occurrence of HFO and the spike waves has higher diagnostic value than the individual occurrence, effectively enhancing surgical efficacy.

Efficacy and safety of bicyclol for treating patients with antituberculosis drug-induced liver injury.

BACKGROUND: Bicyclol was used for treating idiosyncratic acute drug-induced liver injury (DILI) in a phase II trial. This study was aimed at evaluating the efficacy and safety of bicyclol 25 and 50 mg thrice a day (TID) for treating acute DILI caused by anti-TB drugs in the light of the trial results.METHODS: We analysed clinical data of patients with TB drug-induced DILI in the trial database. The primary endpoint was reduction in serum alanine aminotransferase (ALT) levels after 4 weeks of treatment compared to baseline.RESULTS: Overall, 148 patients were included, with respectively 48, 52 and 48 patients included in the control (456 mg polyene phosphatidylcholine TID), high-dose (50 mg bicyclol TID) and low-dose (25 mg bicyclol TID) groups. ALT levels decreased by respectively â-"149.0 (IQR â-"299.3 to â-"98.3 (), â-"225.5 (IQR â-"309.3 to â-"181.8 ) and â-"242.5 (IQR â-"364.8 to â-"153.8) U/L in the control, high-dose and low-dose groups (P < 0.001). The ALT normalisation rates at weeks 1, 2, 4, 6 and 8 were higher in the high- and low-dose groups, while adverse events and serious adverse events were similar across groups.CONCLUSIONS: Bicyclol (25 and 50 mg TID) is effective and safe in treating anti-TB DILI, and bicyclol 50 mg TID showed higher efficacy.

Molecular docking and network pharmacological analysis of Scutellaria baicalensis against renal cell carcinoma.

OBJECTIVE: This paper employs network pharmacology and molecular docking to analyze the active components, targets, and molecular mechanisms of Scutellaria baicalensis in treating renal cell carcinoma (RCC). MATERIALS AND METHODS: The potential active target genes and components of Scutellaria baicalensis are obtained by searching the TCMSP database, and RCC targets are obtained using OMIM, Genecards, and Drugbank databases. The interaction of target proteins is analyzed thanks to STRING, and the component target disease network diagram is constructed through Cytoscape 3.8.2 software. Besides, KEGG, and GO enrichment analysis is performed using the Bioconductor bioinformatics R software package. AutoDock Vina 1.1.2, PyMol 2.5 and Maestro 12.9 software are used for molecular docking. RESULTS: According to the results, Scutellaria baicalensis, which has 36 active ingredients, 500 drug targets, and 85 drug-disease common targets in the treatment of RCC, relies mainly on active ingredients, including wogonin, baicalein, acacetin, oroxylin A, moslosooflavone, salvigenin, and neobaicalein. In addition, the core components within Scutellaria baicalensis that contribute to the treatment of renal cancer are TP53, CCND1, STAT3, CASP3, JUN, VEGFA, AKT1, and EGFR; while the main molecular mechanisms that helps relieve RCC include PI3K-Akt, Ras, MAPK, p53, VEGF, and JAK-STAT signaling pathway. Molecular docking suggested that wogonin had a good binding affinity with core proteins CASP3, CCND1, JUN, STAT3, TP53, and VEGFA. CONCLUSIONS: This study confirms that Scutellaria baicalensis can treat RCC in a multi-component, multi-target, and multi-way manner.

A versatile control program for positioning and shooting targets in laser-plasma experiments.

We introduce a LabVIEW-based control program that significantly improves the efficiency and flexibility in positioning and shooting solid targets in laser-plasma experiments. The hardware driven by this program incorporates a target positioning subsystem and an imaging subsystem, which enables us to install up to 400 targets for one experimental campaign and precisely adjust them in six freedom degrees. The overall architecture and the working modes of the control program are demonstrated in detail. In addition, we characterized the distributions of target positions of every target holder and simultaneously saved the target images, resulting in a large dataset that can be used to train machine learning models and develop image recognition algorithms. This versatile control system has become an indispensable platform when preparing and conducting laser-plasma experiments.

[Taxonomic rank of human parasites].

Biological category is effective to indicate the evolution of organism populations between past and present. Conventional taxonomy of human parasites mainly depends on important morphological features, which suffers from a problem of categorizing related-genera species with similar morphological characteristics. With recent advances in molecular biological technologies, the effective applications of mitochondrial and ribosomal biomarkers and sequencing greatly improve the development of the taxonomic rank of human parasites. Worldwide, the classification of human parasites have been continuously revised and improved. Hereby, we re-categorize parasitic Protozoa, Trematoda, Cestoda and Nematoda, so as to provide insights into the researches on molecular systematics and genetic evolution of human parasites.

[Clinical epidemiological survey of primary liver cancer in Yunnan province from 2005 to 2014].

Objective: To investigate the clinical characteristics and changing trends of primary liver cancer in Yunnan province from 2005 to 2014, in order to provide theoretical basis for the prevention and treatment of liver cancer in this region. Methods: A retrospective survey was used to select inpatient cases of liver cancer who were initially diagnosed and treated in our hospital from 2005 to 2014 with simple random sampling. Patients socio-demographic and clinicopathological characteristics were extracted by a unified and standardized questionnaire, and the data were statistically analyzed. Results: A total of 1000 cases with liver cancer were included, aged (53.2±11.2) years, with a male-to-female ratio of 5.99/1.00. There was no significant change in the gender and age composition ratio of patients in the past 10 years. The proportion of patients with lower education level (primary or junior high school) were increased from 21.8% to 23.4%, and the proportion of patients with relatively higher education level were decreased from 58% to 38.2% (P<0.001). Smokers and non-smokers patients were decreased and increased from 58.8% to 44.4%, and 41.2% to 55.6% (P<0.001). The proportion of drinker patients were decreased from 46.4% to 35.2%. The proportion of patients with advanced liver cancer (stage C and D) were increased, while the proportion of patients with stage A and B showed a downward trend (P<0.001). The proportion of HBsAg-positive patients showed an upward trend, that is, rising from 69% in 2005 to 82% in 2014 (P=0.043). The proportion of HBeAg-positive patients showed a steady trend (P=0.008). The use rate of ultrasound examination in patients with liver cancer were decreased from 91.0% to 58.0% (P=0.001), while the use rate of computed tomography (CT), MRI, and PET/CT examinations were increased from 81.0% to 84.0% (P=0.05), 0 to 22% (P<0.001), and 0 to 3% (P=0.026) between 2005 to 2014. The proportion of surgical patients were increased (P=0.005), but the proportion of interventional patients did not change significantly (P=0.590). Surgery and interventional therapy were the most common treatment methods, and the proportion of patients treated with surgery over the past 10 years showed an upward trend (P=0.005), while the proportion of interventional therapy remained at a high level with no significant change (P=0.590). Conclusion: In Yunnan province, the incidence of liver cancer increases with age, and the proportion of male with liver cancer is almost six times that of women. Moreover, the low positive rate of alpha-fetoprotein levels and advanced clinical stage in this region are presently the main challenges against the liver cancer prevention and treatment. The application scope of CT, magnetic resonance imaging, PET-CT and other examination methods has gradually expanded, but the treatment methods are still mainly surgery and interventional therapy.

[Diagnostic efficacy of indirect haemagglutination assay for detection of Schistosoma japonicum infections among boatmen and fishermen in the Dongting Lake region].

OBJECTIVE: To evaluate the diagnostic efficacy of indirect haemagglutination assay (IHA) for detection of Schistosoma japonicum infections among boatmen and fishermen in Dongting Lake region, so as to provide insights into improving the schistosomiasis surveillance program among boatmen and fishermen. METHODS: The boatmen and fishermen were detected for S. japonicum infections using IHA and Kato-Katz technique or miracidium hatching test nylon gauze simultaneously at schistosomiasis testing sites in the anchor sites for boatmen and fishermen in the Dongting Lake region during the period from 2014 to 2016, and using IHA for serological screening followed by parasitological testing of seropositives during the period from 2017 to 2019. The sensitivity and specificity of IHA were evaluated for detection of S. japonicum infections among boatmen and fishermen, with the 2014-2016 parasitological testing results as a gold standard. In addition, the seroprevalence of S. japonicum infections was compared among boatmen and fishermen with different characteristics and among years. RESULTS: A total of 306 schistosomiasis testing sites were assigned for boatmen and fishermen, and a total of 143 360 person-time boatmen and fishermen were tested for S. japonicum infections in the Dongting Lake region from 2014 to 2019. The sensitivity and specificity of IHA were 69.9%, 97.3% and 96.1% (χ2 = 74.6, P < 0.05), and 70.9%, 74.5% and 71.9% for detection of S. japonicum infections from 2014 to 2016 (χ2 = 29.4, P < 0.05), respectively. The seroprevalence of S. japonicum infections reduced from 30.3% in 2014 to 1.8% in 2019 among boatmen and fishermen, appearing an overall tendency towards a decline (Z = 1 552.4, P < 0.05). In addition, male, individuals at ages of 45 to 60 years, full-time boatmen and fishermen were more likely to be seropositive for S. japonicum infections (all P values < 0.05). CONCLUSIONS: The seroprevalence of S. japonicum infections appeared a tendency towards a decline among boatmen and fishermen in the Dongting Lake region year by year from 2014 to 2019. IHA presented a high efficacy for screening of S. japonicum infections among boatmen and fishermen in the Dongting Lake region.

Identification of SCAR markers for genetic authentication of Dendrobium nobile Lindl.

Dendrobium nobile Lindl. is an orcid plant with important medicinal values. This is a colourful houseplant, and also a popular herb in traditional Chinese medicine (TCM). The variants of this plant from different geographic regions might be high, and in this study, we aimed to develop specific sequence characterized amplified region (SCAR) markers for the identification of specific variant of this plant. Different cultivars of D. nobile were collected from nine different places of China, and one cultivar from Myanmar. DNA materials were extracted from the plant samples, random amplified polymorphic DNA (RAPD) were developed, cloned and sequenced for the development of SCAR markers. We have developed four SCAR markers, which are specific to the cultivar from Luzhou China, and clearly distinguishable (genetically) from other cultivars. These SCAR markers are deposited in GenBank (accession number MZ417502, MZ484089, MZ417504 and MZ417505). Four SCAR markers for D. nobile are effective molecular technique to genetically identify the different cultivars or species, and this method is applicable for genetic characterization and identification of other plant species too.

[Comparative study on fracture toughness of digital light processing three-dimensional printing zirconia and milled zirconia].

Objective: To investigate the influence of digital light processing (DLP) and computer numerical controlmilling (CNC) on the mechanical behavior of zirconia. Methods: Prepared DLP samples (experimental group, n=52) and CNC samples (control group, n=52) with 12 samples in each group were randomly selected using random number table to measure density, grain size and crystal phase composition. According to the different methods fracture toughness test, the samples were divided into indentation method group (IM) and single-edge-V-notch-beam group (SEVNB), with 30 DLP and 30 CNC samples in IM group, 10 DLP and 10 CNC samples in SEVNB group. The IM group was tested under three different loads (49.03 N, 98.07 N, 196.10 N), there were 10 samples for each load and each sample was tested at 15 points, and the load with the ratio of crack length to indentation diagonal length greater than 2.5 was selected as the indentation load to calculate its IM fracture toughness. At the same time, the SEVNB group was tested with four point bending test to record the maximum load at the time of fracture and calculate the SEVNB fracture toughness. Finally, the indentation and fracture surface were observed using optical microscope and scanning electron microscope, and the results of DLP group and CNC group were further compared to explore the difference in fracture mechanism. Results: The microstructure of DLP and CNC zirconia was basically the same, the density of DLP group was (6.020±0.021) g/cm3, the grain size was (0.603±0.033) µm; the density of CNC group was (6.038±0.012) g/cm3, the grain size was (0.591±0.033) µm. Both groups were composed of tetragonal zirconia. The load of 196.10 N was chosen as the indentation load for two groups to calculate the IM fracture toughness. In terms of fracture toughness, there was no significant difference between the two groups (P>0.05). Scanning electron microscope images of fracture surface showed the intergranular fracture was the leading fracture mode of two groups. The IM and SEVNB fracture toughness of DLP zirconia were (6.111±0.179) MPa·m1/2 and (7.221±0.809) MPa·m1/2, respectively. The IM and SEVNB fracture toughness of CNC zirconia were (6.126±0.383) MPa·m1/2 and (7.408±0.533) MPa·m1/2, respectively. Conclusions: The microstructure of DLP and CNC zirconia is almost the same, and there is little difference in the fracture toughness of zirconia between two processing technologies.

Increased risk of diabetes in cancer survivors: a pooled analysis of 13 population-based cohort studies.

BACKGROUND: Diabetes is considered as an established risk factor for cancer development. However, the link between diabetes among cancer survivors remains inconclusive. The hypothesis of this study was to assess the hazard ratio (HR) of incidence of diabetes in cancer survivors compared with the HR in the general population. PATIENTS AND METHODS: A comprehensive literature search was performed in PubMed, Embase, and the Cochrane Library from database inception to 15 December 2020 for population-based cohort studies. Summary effect estimates were combined using random-effects models. We also performed subgroup analyses to test sources of heterogeneity and the stability of the results stratified by various study and participant characteristics. RESULTS: Thirteen population-based cohort studies involving 1 686 595 participants were analyzed. The HR for the development of diabetes in cancer survivors was 1.39 [95% confidence interval (CI) 1.29-1.50; I2 = 82.3%; P < 0.001] compared with that in noncancer controls, among which survivors of hematological, gynecologic, breast, colorectal and urinary tract cancer (all P < 0.05) showed consistent significant results, whereas no significant increased risk was observed for other cancer types. The effects were more prominent in populations of shorter cancer survival duration (<1 year) (HR 2.09, 95% CI 1.32-3.32; P = 0.009). Moreover, cancer survivors with a longer follow-up period (>10 years) had a relatively higher risk of diabetes (HR 1.54, 95% CI 1.34-1.77) than those with a shorter follow-up period. CONCLUSIONS: In this large pooled analysis of population-based cohorts, evidence supports the hypothesis that the risk of developing diabetes is increased in cancer survivors compared with the general population. We should interpret the results with caution for considerable interstudy heterogeneity. However, health policy makers should take this as a challenge for the early prevention and effective intervention of diabetes.

Early outcome and safety of bedaquiline-containing regimens for treatment of MDR- and XDR-TB in China: a multicentre study.

OBJECTIVES: Bedaquiline treatment significantly improves multidrug-resistant tuberculosis (MDR-TB) patient treatment outcomes. However, safety and efficacy data are lacking for bedaquiline used with background regimens to treat Chinese TB patients. Here, we describe our initial clinical experience for bedaquiline treatment of a large multicentre cohort of MDR-TB and extensively drug-resistant tuberculosis (XDR-TB) patients in China. METHODS: Patients (177) received 24-week bedaquiline treatment combined with personalized anti-TB drug background regimens. As primary efficacy endpoints, times to initial sputum culture conversion were measured. RESULTS: Of 177 MDR-TB patients completing the 24-week treatment course, sputum culture conversion occurred for 151/177 (85.3%), while 26 had unfavourable outcomes, including 3/177 (1.7%) deaths and 23/177 (13.0%) non-responders at treatment completion. The median time to sputum culture conversion was 4 (interquartile range 2-8) weeks. Conversion rates were 33/39 (84.6%, 95% confidence interval (CI) 73.3-95.9) for MDR-TB patients, 47/56 (83.9%, 95% CI 74.3-93.6) for pre-XDR-TB patients and 71/82 (86.6%, 95% CI 79.2-94.0) for XDR-TB patients. Multivariate analysis demonstrated that patients with low body mass index (odds ratio 7.356; 95% CI 2.652-20.401) were at significantly high risk of unfavourable outcomes, with serious adverse events noted in 15 (8.5%) patients, including six with corrected QT interval (QTc) prolongation times (>500 ms). CONCLUSION: Bedaquiline, when included in background regimens for treatment of MDR-TB and XDR-TB patients in China, was safe and associated with a high rate of culture conversion.

Monte Carlo implementation of new algorithms for the evaluation of averaged-dose and -track linear energy transfers in 62 MeV clinical proton beams.

We exploited the power of the Geant4 Monte Carlo toolkit to study and validate new approaches for the averaged linear energy transfer (LET) calculation in 62 MeV clinical proton beams. The definitions of the averaged LET dose and LET track were extended, so as to fully account for the contribution of secondary particles generated by target fragmentation, thereby leading to a more general formulation of the LET total. Moreover, in the proposed new strategies for the LET calculation, we minimised the dependencies in respect to the transport parameters adopted during the Monte Carlo simulations (such as the production cut of secondary particles, voxel size and the maximum steplength). The new proposed approach was compared against microdosimetric experimental spectra of clinical proton beams, acquired at the Italian eye proton therapy facility of the Laboratori Nazionali del Sud, Istituto Nazionale di Fisica Nucleare (INFN-LNS, Catania, I) from two different detectors: a mini-tissue equivalent proportional chamber (TEPC), developed at the Legnaro National Laboratories of the National Institute for Nuclear Physics (LNL-INFN) and a silicon-on-insulator (SOI) microdosimeter with 3D sensitive volumes developed by the Centre for Medical Radiation Physics of Wollongong University (CMRP-UoW). A significant increase of the LET in the entrance region of the spread out Bragg peak (SOBP) was observed, when the contribution of the generated secondary particles was included in the calculation. This was consistent with the experimental results obtained.

[Analysis of the clinicopathologic features as well as diagnosis and treatment of 59 patients with Castleman disease].

Objective: To investigate the clinicopathologic features, treatment, and prognosis in patients with Castleman disease (CD) . Methods: We retrospectively analyzed the clinicopathologic data of 59 patients for whom a diagnosis of Castleman disease was confirmed using pathological examination from October 2011 to October 2019 at the Henan Cancer Hospital. The patients were divided into the following two groups as per the following clinical classifications: unicentric CD (UCD, n=47) and multicentric CD (MCD, n=12) . Data on clinical manifestations, laboratory findings, treatment, and prognosis were analyzed. Results: There was no significant difference in the median age and the ratio of male to female between the UCD and MCD. UCD was characterized by asymptomatic enlargement of the single lymph node. The main pathological type was hyaline vascular histopathology (83.0%) . Of these, 44 patients chose surgical resection, and their prognosis was good. Treatment. MCD was characterized by multiple enlarged superficial and/or deep lymph nodes with B symptoms, weakness, and hepatosplenomegaly. Anemia, hypoproteinemia, and globulin level were increased on laboratory examinations. Plasmacyte histopathology was the main pathological type and was present in about 50.0% of the subjects. Only chemotherapy was performed for these MCD patients, followed by chemotherapy or chemotherapy followed by radiotherapy, and the efficient was 58.3% (7/12) . Conclusions: UCD, characterized by asymptomatic enlargement of the single lymph node, shows good postoperative prognosis. MCD has relatively complex clinical manifestations and poor prognosis, and optimal treatment is yet to be established.

[Effect and mechanism of adipocyte co-culture on aquaporin-9 expression in HepG2 cells].

Objective: To observe the effect of differentiated mature adipocytes on hepatic steatosis and aquaporin-9 (AQP9) expressions in HepG2 cells and further explore its possible mechanism of action. Methods: Human preadipocytes were cultured and differentiated to full maturity. HepG2 cells were co-cultured with non-differentiated adipocytes and differentiated mature adipocytes for 48 h, and then labeled as control group and experimental group. Oil red O staining and intracellular triglyceride content were performed on co-cultured HepG2 cells and simultaneous changes in phosphatidylinositol 3-kinase (PI3K) - serine/threonine kinase (Akt) signaling pathway, and AQP9 mRNA and protein levels were detected. The experimental group was co-cultured with recombinant human insulin-like growth factor-I (IGF-I), with the addition of 100ng/ml PI3K-Akt pathway agonist, labeled as experimental group + IGF-I group. The activation of PI3K-Akt pathway was verified by Western blotting (WB). The expression of AQP9 was detected by RT-q PCR and WB. The recombinant lentivirus LV-AQP9 or empty-loaded virus LV-PWPI was transfected with HepG2 cells by recombinant lentiviral transfection tecnique, and labeled as HepG2-AQP9 and HepG2-PWPI. The transfection efficiency was assessed by confocal laser scanning microscopy and RT-qPCR and WB detected the change of AQP9 expression level after virus transfection. Afterwards, the stable over-expressed HepG2-AQP9 cells and the empty-loaded HepG2-PWPI cells were co-cultured with differentiated mature adipocytes for 48h, and labeled as HepG2-AQP9 co-culture group, and then intracellular triglyceride content were detected with Oil red O staining. Finally, IGF-I was added to the HepG2-AQP9 co-culture group, which was recorded as HepG2-AQP9 co-culture + IGF-I group. Intracellular triglyceride content was detected with Oil red O staining, and WB verified PI3K-Akt signaling pathway activation and changes in AQP9 mRNA and protein levels. A t-test was used to compare the two independent samples. Results: The intracellular lipid droplets and triglyceride content (0.052 ± 0.005) in the experimental group was increased significantly than the control group (0.033 ± 0.003) (t= 5.225,P= 0.006), suggesting that adipocyte co-culture had induced steatosis in HepG2 cells. RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (3.615 ± 0.330) and protein levels (0.072 ± 0.005) in the experimental group were significantly higher than the control group (t= 13.708, 11.225,P= 0.005, < 0.001). WB results showed that the expression level of phosphorylated Akt (p-Akt) protein (0.116±0.003) in the experimental group was significantly lower than the control group (0.202 ± 0.003) (t= 27.136,P< 0.001). The total Akt protein was constant, and the p-Akt/total Akt (0.182 ± 0.017)was significantly lower than the control group (0.327 ± 0.019) (t= 2.431,P= 0.001), suggesting that adipocyte co-culture had inhibited PI3K- Akt signaling pathway in HepG2 cells and up-regulated the expression level of AQP9. WB results indicated that the expression level of p-Akt protein (0.194 ± 0.021) in the experimental group + IGF-I group was significantly higher than the experimental group (0.132 ± 0.003) (t= 5.082,P= 0.007). The total Akt protein was constant, and the p-Akt/total Akt (0.281 ± 0.009) was significantly higher than the control group (0.184 ± 0.132) (t= 10.311,P< 0.001). Simultaneously, RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (0.327 ± 0.347) and protein levels (0.042 ± 0.004) in the experimental group + IGF-I group were significantly lower than the experimental group (t= 33.573, 5.598,P< 0.001, 0.005), suggesting that adipocyte co-culture had possibility to regulate the expression level of AQP9 through the PI3K-Akt pathway. Confocal laser microscopy analysis showed that the transfection efficiency was more than 90%. RT-q PCR and WB results indicated that the expression levels of AQP9 mRNA and protein levels (0.373 ± 0.221) in HepG2-AQP9 group were significantly higher than HepG2-PWPI group (t=14.953, 28.931,P= 0.002 and 0.000), suggesting that the stable overexpression of AQP9 cell line was successfully constructed. The intracellular lipid droplets and triglyceride content in HepG2-AQP9 co-culture group was significantly increased (t= 5.478, 5.369,P= 0.005) than HepG2-PWPI co-culture group and HepG2-AQP9 co-culture+ IGF-I group, suggesting that the increased expression of AQP9 had promoted HepG2 steatosis in co-cultured adipocytes. WB results showed the expression levels of p-Akt protein (0.168 ± 0.006) and p-Akt/total Akt (0.265±0.009) in HepG2-AQP9 co-culture + IGF-1 group was significantly increased (t= 16.311, 8.769,P< 0.001) than HepG2-AQP9 co-culture group, while the expression levels of AQP9 mRNA (0.327 ± 0.034) and protein (0.375 ± 0.025) was significantly decreased (t= 33.573, 9.146,P< 0.001 and 0.001). Conclusion: Adipocytes co-culture can induce steatosis in HepG2 cells, and may participate in inhibiting PI3K-Akt signaling pathway to upregulate the expression of AQP9 in steatotic HepG2 cells.

[Application of multi-slice spiral CT to assess vascular volume for the diagnosis and treatment of gastric varices].

Objective: To differentiate the inflow and outflow channels of gastric varices in cirrhotic portal hypertension patients using multi-slice spiral CT (MSCT), and to assess the relationship between calculable CT volume of gastric varices and the amount of tissue adhesive. Methods: 97 cases with cirrhotic gastric varices who were admitted from November 2013 to August 2017 were selected. The type and shape of gastric varices were observed before tissue glue injection treatment by MSCT. The correlation between CT volume of gastric varices and the amount of tissue adhesive was evaluated by Spearman rank correlation coefficient and Univariate linear regression analysis. Results: MSCT showed that Le, g type had the highest proportion (54.6%), followed by Le, g, Lg (20.6%). Le, Lg and Lgf type accounted for 17.5%, and 5.2%, respectively, while Lgf+b accounted for 2.1%. On MSCT, varices of the gastric fundus were in the direction from bottom to top, and 75% of the fundus had a large curved side varices combined with gastric and renal shunt. Under the gastroscopy, varices in the small curved side of the gastric fundus from near to far were formless. In addition, varices in the large curved side of the gastric fundus when observed from different angles to the direction of blood flow (reverse gastroscope) were 72.7% (near and far) or 20.5 % (far and near). There was a positive correlation between CT volume (R = 0.97, P < 0.001) and the amount of tissue adhesive (Y(1) = 0.35 + 0.65X1, Univariate linear regression equation; ρ = 0.89, P < 0.001, Spearman correlation analysis). Conclusion: MSCT can recognize the vascular shape and inflow and outflow channels of gastric varices. A positive correlation between CT volume and the amount of tissue adhesive, suggested that the CT volume measurement before treatment could be used as one of the method to predict the amount of tissue adhesive.

Non-invasive prenatal testing for fetal inheritance of maternal ß-thalassaemia mutations using targeted sequencing and relative mutation dosage: a feasibility study.

OBJECTIVE: To evaluate whether targeted sequencing and relative mutation dosage can be used to diagnose correctly inheritance of maternal ß-thalassaemia mutations in cell-free DNA. DESIGN: Feasibility study using samples collected in a prenatal clinic. SETTING: South East Asia. POPULATION: Couples where both partners were known to be carriers of one of four common ß-thalassaemia mutations or an HbE mutation, and therefore at risk of carrying a fetus affected with ß-thalassaemia. METHODS: 49 samples previously identified as having inherited a paternal ß-thalassaemia mutation were amplified using nested polymerase chain reaction (PCR), and then sequencing. Relative mutation dosage was used to classify the fetus as having inherited the wild-type or mutant maternal allele. MAIN OUTCOME MEASURES: Classification of the fetus as 'unaffected' (if the maternal wild-type allele was inherited) or 'affected' with ß-thalassaemia (if the maternal mutant allele was inherited). RESULTS: A classification for inheritance of maternal allele was obtained in 48/49 samples (98.0%). A concordant call was made in 44/48 cases (91.7%): one false-positive and three false-negatives were obtained. Thus, we had an overall sensitivity of 87.5% [95% confidence interval (CI) 67.6-97.3%] and a specificity of 95.8% (95% CI 78.9-99.9%) for inheritance of maternal genotype. CONCLUSIONS: RMD for detection of inheritance of maternal ß-thalassaemia mutations has potential for clinical use. Our sequential approach could be applied to other single-gene disorders. TWEETABLE ABSTRACT: NIPT for ß-thalassaemia achieved using nested-PCR followed by relative mutation dosage.