Phenotype and genotype analyses of two pedigrees with inherited fibrinogen deficiency / 中华血液学杂志

Objective: To analyze the phenotype and genotype of two pedigrees with inherited fibrinogen (Fg) deficiency caused by two heterozygous mutations. We also preliminarily probed the molecular pathogenesis. Methods: The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and plasma fibrinogen activity (Fg∶C) of all family members (nine people across three generations and three people across two generations) were measured by the clotting method. Fibrinogen antigen (Fg:Ag) was measured by immunoturbidimetry. Direct DNA sequencing was performed to analyze all exons, flanking sequences, and mutated sites of FGA, FGB, and FGG for all members. Thrombin-catalyzed fibrinogen polymerization was performed. ClustalX 2.1 software was used to analyze the conservatism of the mutated sites. MutationTaster, PolyPhen-2, PROVEAN, SIFT, and LRT online bioinformatics software were applied to predict pathogenicity. Swiss PDB Viewer 4.0.1 was used to analyze the changes in protein spatial structure and molecular forces before and after mutation. Results: The Fg∶C of two probands decreased (1.28 g/L and 0.98 g/L, respectively). The Fg∶Ag of proband 1 was in the normal range of 2.20 g/L, while it was decreased to 1.01 g/L in proband 2. Through genetic analysis, we identified a heterozygous missense mutation (c.293C>A; p.BβAla98Asp) in exon 2 of proband 1 and a heterozygous nonsense mutation (c.1418C>G; p.BβSer473*) in exon 8 of proband 2. The conservatism analysis revealed that Ala98 and Ser473 presented different conservative states among homologous species. Online bioinformatics software predicted that p.BβAla98Asp and p.BβSer473* were pathogenic. Protein models demonstrated that the p.BβAla98Asp mutation influenced hydrogen bonds between amino acids, and the p.BβSer473* mutation resulted in protein truncation. Conclusion: The dysfibrinogenemia of proband 1 and the hypofibrinogenemia of proband 2 appeared to be related to the p.BβAla98Asp heterozygous missense mutation and the p.BβSer473* heterozygous nonsense mutation, respectively. This is the first ever report of these mutations.

Multiple myeloma presenting as acute tubulointerstitial nephritis

Background Acute tubulointerstitial nephritis (ATIN) is a very rare paraneoplastic manifestation in patients with multiple myeloma (MM). It is an uncommon pattern of renal disease in such patients. Case presentation We report a case of an 82-year-old male who was admitted with acute kidney injury. Renal biopsy showed typical findings of light chain-associated ATIN with scattered inflammatory cells in the interstitium and associated active tubulitis. No other common manifestations of MM were present at the time of presentation, including hypercalcemia, hyperuricemia, proteinuria, bone pain or lytic bone lesions. Subsequent immunoassays revealed significant serum lambda light chain burden and Bence Jones protein in urine. Immunofluorescence demonstrated linear tubular basement membranes with positive staining for lambda light chain (3+). Electron microscopy (EM) further showed interstitial edema and inflammation. All the aforementioned findings are consistent with ATIN and supported the diagnosis of MM. Conclusions In conclusion, light chain-associated ATIN should be considered in the differential diagnosis of acute interstitial nephritis. Henceforth, serum free light chains as well as serum and urine protein electrophoresis should be included in the workup of such patients.

The Distinct Role of Tcfs and Lef1 in the Self-Renewal or Differentiation of Mouse Embryonic Stem Cells

Background and Objectives@#Tcfs and Lef1 are DNA-binding transcriptional factors in the canonical Wnt signaling pathway. In the absence of β-catenin, Tcfs and Lef1 generally act as transcriptional repressors with co-repressor proteins such as Groucho, CtBP, and HIC-5. However, Tcfs and Lef1 turn into transcriptional activators during the interaction with β-catenin. Therefore, the activity of Tcfs and Lef1 is regulated by β-catenin. However, the intrinsic role of Tcfs and Lef1 has yet to be examined. The purpose of this study was to determine whether Tcfs and Lef1 play differential roles in the regulation of self-renewal and differentiation of mouse ES cells. @*Methods@#and Results: Interestingly, the expression of Tcfs and Lef1 was dynamically altered under various differentiation conditions, such as removal of LIF, EB formation and neuronal differentiation in N2B27 media, suggesting that the function of each Tcf and Lef1 may vary in ES cells. Ectopic expression of Tcf1 or the dominant negative form of Lef1 (Lef1-DN) contributes to ES cells to self-renew in the absence of leukemia inhibitory factor (LIF), whereas ectopic expression of Tcf3, Lef1 or Tcf1-DN did not support ES cells to self-renew. Ectopic expression of either Lef1 or Lef1-DN blocked neuronal differentiation, suggesting that the transient induction of Lef1 was necessary for the initiation and progress of differentiation. ChIP analysis shows that Tcf1 bound to Nanog promoter and ectopic expression of Tcf1 enhanced the transcription of Nanog. @*Conclusions@#The overall data suggest that Tcf1 plays a critical role in the maintenance of stemness whereas Lef1 is involved in the initiation of differentiation.

Comparative transcriptomic analysis of two important life stages of Angiostrongylus cantonensis: fifth-stage larvae and female adults

Abstract The mechanisms involved in the fast growth of Angiostrongylus cantonensis from fifth-stage larvae (L5) to female adults and how L5 breaks through the blood-brain barrier in a permissive host remain unclear. In this work, we compared the transcriptomes of these two life stages to identify the main factors involved in the rapid growth and transition to adulthood. RNA samples from the two stages were sequenced and assembled de novo. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of 1,346 differentially expressed genes between L5 and female adults was then undertaken. Based on a combination of analytical results and developmental characteristics, we suggest that A. cantonensis synthesizes a large amount of cuticle in L5 to allow body dilatation in the rapid growth period. Products that are degraded via the lysosomal pathway may provide sufficient raw materials for cuticle production. In addition, metallopeptidases may play a key role in parasite penetration of the blood-brain barrier during migration from the brain. Overall, these results indicate that the profiles of each transcriptome are tailored to the need for survival in each developmental stage.

Establishment and validation of improved six-year-old pediatric thorax human model / 医用生物力学

Objective To improve the biological fidelity of the thorax flexible body in the original MADYMO child human model,so as to further study pediatric thorax injuries of child occupant.Methods The finite element model of six-year-old pediatric thorax was built by the method of reverse modeling based on CT images.By replacing the thorax model with flexible body in MADYMO six-year-old human model,an improved human model containing biomechanical thorax model was developed.The model was verified by joint validation of two tests,including Irwin and Mertz's method of scaling channel reported in Kroell's adult chest impact experiment and Ouyang's thoracic impact test on pediatric cadavers.Results The response of this established thorax model was in good agreement with scaling channel method and cadaver test data,and the thorax model was much more accurate than the original flexible body model.The resilience of simulation model was consistent with cadaver test.Conclusions The validity of the model is verified,and the results can be further used for occupant injury analysis in vehicle frontal crash.

Establishment and validation of improved six-year-old pediatric thorax human model / 医用生物力学

Objective To improve the biological fidelity of the thorax flexible body in the original MADYMO child human model,so as to further study pediatric thorax injuries of child occupant.Methods The finite element model of six-year-old pediatric thorax was built by the method of reverse modeling based on CT images.By replacing the thorax model with flexible body in MADYMO six-year-old human model,an improved human model containing biomechanical thorax model was developed.The model was verified by joint validation of two tests,including Irwin and Mertz's method of scaling channel reported in Kroell's adult chest impact experiment and Ouyang's thoracic impact test on pediatric cadavers.Results The response of this established thorax model was in good agreement with scaling channel method and cadaver test data,and the thorax model was much more accurate than the original flexible body model.The resilience of simulation model was consistent with cadaver test.Conclusions The validity of the model is verified,and the results can be further used for occupant injury analysis in vehicle frontal crash.

Establishment and validation of improved six-year-old pediatric thorax human model / 医用生物力学

Objective To improve the biological fidelity of the thorax flexible body in the original MADYMO child human model,so as to further study pediatric thorax injuries of child occupant.Methods The finite element model of six-year-old pediatric thorax was built by the method of reverse modeling based on CT images.By replacing the thorax model with flexible body in MADYMO six-year-old human model,an improved human model containing biomechanical thorax model was developed.The model was verified by joint validation of two tests,including Irwin and Mertz's method of scaling channel reported in Kroell's adult chest impact experiment and Ouyang's thoracic impact test on pediatric cadavers.Results The response of this established thorax model was in good agreement with scaling channel method and cadaver test data,and the thorax model was much more accurate than the original flexible body model.The resilience of simulation model was consistent with cadaver test.Conclusions The validity of the model is verified,and the results can be further used for occupant injury analysis in vehicle frontal crash.

Recent progress in synthesis and pharmacological effects of ellipticine and its derivatives / 药学学报

Ellipticine is an alkaloid isolated from natural product with cytotoxicity, which has antitumor and anti-aids activity. Since it was first identified in 1959, a great deal of effort has been devoted to the development of various approaches for synthesis of ellipticine. This review provides a summary for synthesis approaches of ellipticine from different starting materials. The antitumor mechanism and structure-activity relationship are also discussed.

Establishment and validation of improved six-year-old pediatric thorax human model / 医用生物力学

Objective To improve the biological fidelity of the thorax flexible body in the original MADYMO child human model, so as to further study pediatric thorax injuries of child occupant. Methods A finite element model of the six-year-old pediatric thorax was built by adopting the method of reverse modeling based on CT images. By replacing the thorax model with flexible body in MADYMO six-year-old human model, an improved human model containing biomechanical thorax model was developed. The model was verified by joint validation of two tests, including Kroell’s adult chest impact experiment combined with Irwin and Mertz’s scaling method, and Jun Ouyang’s thoracic impact test on pediatric cadavers. Results The response of this established thorax model was in good agreement with scaling method and cadaver test data, and the thorax model was much more accurate than the original flexible body model. The resilience of simulation model was consistent with cadaver test. Conclusions The validity of the model is verified, and can be further used in occupant injury analysis in vehicle frontal crash.

Clinical outcomes Tight rope versus traditional screw fixation for the treatment of injury of distal tibiofibular syndesmosis in ankle fracture / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effectiveness for Tight rope fixation and traditional screw fixation in treating injury of distal tibiofibular syndesmosis in ankle fractures.</p><p><b>METHODS</b>A retrospective study was carried out in patients with injury of distal syndesmosis in ankle fractures who received 2 surgical operations(observation group: 33 cases with Tight rope fixation; control group: 35 cases with traditional screw fixation) from May 2014 to February 2016. There were 18 males and 15 females, aged from 20 to 55 years old with an average of(32.4±5.2) years old in observation group; of them, 19 cases were caused by traffic accidents, 10 by sprain, and 4 by falling; according to Lauge-Hansen typing of ankle fractures, all of the 33 cases were pronation-extorsion fracture, 12 cases were III degree and 21 cases were IV degree. There were 19 males and 16 females, aged from 21 to 54 years old with an average of (32.8±5.5) years old in control group; of them, 20 cases were caused by traffic accidents, 11 by sprain, 4 by falling; according to Lauge-Hansen typing of ankle fractures, 1 case was with pronation-outreach, 34 cases with pronation-extorsion, 13 cases were III degree and 21 cases were IV degree. Fixation time and complication were compared between two groups and AOFAS scores were observed in two groups 3 and 6 months after the operation as well as final follow-up.</p><p><b>RESULTS</b>All the patients were followed up from 8 to 24 months with an average of (16.3±3.8) months. Fixation time of observation group and control group were (10.1±2.8) min and (9.5±2.3) min(>0.05) respectively. There were significant difference in complication and AOFAS of 3, 6 months postoperatively between two groups(<0.05). In observation group, 23 case got excellent result, 9 good, 1 fair; and in control group, 18 cases got excellent results, 12 good, 5 fair; there was no significant difference between two groups(>0.05).</p><p><b>CONCLUSIONS</b>Tight rope for the treatment of injury of distal tibiofibular ligament union in ankle fractures has advantages such as easier techniques, earlier weight-bearing exercises without risk of screw breakage, thus is a new choice. However, it is still necessary to further study the angle, direction and tension of button steel plate.</p>

Efficient derivation of functional hepatocytes from mouse induced pluripotent stem cells by a combination of cytokines and sodium butyrate / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hepatocyte transplantation has been proposed as an alternative to whole-organ transplantation to support many forms of hepatic insufficiency. Unfortunately, the lack of donor livers makes it difficult to obtain enough viable human hepatocytes for hepatocyte-based therapies. Therefore, it is urgent to find new ways to provide ample hepatocytes. Induced pluripotent stem (iPS) cells, a breakthrough in stem cell research, may terminate these hinders for cell transplantation. For the promise of iPS cells to be realized in liver diseases, it is necessary to determine if and how efficient they can be differentiated into functional hepatocytes.</p><p><b>METHODS</b>In this study, we directly compared the hepatic-differentiation capacity of mouse iPS cells and embryonic stem (ES) cells with three different induction approaches: conditions via embryonic body (EB) formation plus cytokines, conditions by combination of dimethyl sulfoxide and sodium butyrate and chemically defined, serum free monolayer conditions. Among these three induction conditions, more homogenous populations can be promoted under chemically defined, serum free conditions. The cells generated under these conditions exhibited hepatic functions in vitro, including glycogen storage, indocynine green (ICG) uptake and release as well as urea secretion. Although efficient hepatocytes differentiation from mouse iPS cells were observed, mouse iPS cells showed relatively lower hepatic induction efficiency compared with mouse ES cells.</p><p><b>RESULTS</b>Mouse iPS cells would be efficiently differentiated into functional hepatocytes in vitro, which may be helpful in facilitating the development of hepatocytes for transplantation and for research on drug discovery.</p><p><b>CONCLUSION</b>We demonstrate that mouse iPS cells retain full potential for fetal liver development and describe procedures that facilitates the efficient generation of highly differentiated human hepatocyte-like cells from iPS cells in vitro.</p>

Isolation and characterization of an iridovirus from sick giant salamander (Andrias davidianus) / 病毒学报

A virus was isolated from cultured sick giant salmander (Andrias davidianus ) in a farm, Shanxi Province, China. Skin ulceration and necrosis of the distal limbs are main clinical symptoms. Virus propagated and caused CPE at 10 degrees C to 30 degrees C in BF-2, CO, CHSE, FHM cells. The optimum condition of replication was in BF-2 cells at 25 degrees C. The virus was proved to be senstive to chloroform, heat, pH3 and pH10 treatment. Viral replication was inhibited by 5-Fluoro-2-deoxyuridine (FUDR). These results indicated that the virus possessed an envelope and DNA as the genome. Electron-microscopic observation of thin-section showed numerous hexagonal viral particles measuring 130 nm to 150 nm in diameter orderly arranged in a lattice form in cytoplasm of BF-2 cells. The particles showed typical iridovirus morphology. A 413 bp fragment was amplified from the viral main capsid protein gene by PCR. The fragments was sequenced and analysed. The results showed the isolate shared more than 96% nucleotide identity with some Ranaviruses. We suggested that this virus was named as Andrias davidianus iridovirus (ADIV) tentatively.

Comparison of characteristics of SVCV strains isolated in China and in Europe / 病毒学报

In the nationwide epidemiological investigation, SVCV-741 was for the first time isolated in Beijing region, China in 2003, and designated as SVCV Asian strain. In this paper, we compared SVCV-741 (Asian strains isolated in China) with SVCV-10/3 (Europe reference strain) on their physico-chemical, biological and morphological characteristics. The results indicated that there were no distinct differences between two SVCV strains on phycico-chemical and morphological characteristics. The main existing differences were: (1) The stability of SVCV-741 to temperature in cell culture was higher than that of SVCV-10/3, which might have some evolutionary and biological implication of SVCV; (2) No SVC outbreak ever occurred caused by SVCV-741;Furthermore we found that both SVCV-741 and SVCV-10/3 grew faster and produced higher virus titer in CO cells than other cell lines. It indicated that CO cell lines might be useful tool for SVCV research.

Establishment and assessment of Caco-2 cell in vitro absorption model / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish and assess the Caco-2 cell in vitro absorption model.</p><p><b>METHODS</b>Caco-2 cells were cultured on the millipore filters fixed in Snapwell transport chamber. The cell morphology, transepithelial electrical resistance, mannitol efflux rate and alkaline phosphatase activities were monitored during culture.</p><p><b>RESULTS</b>After 21 days of in vitro culture, formation of tight junction was observed between the cells. The transepithelial electrical resistance reach a relatively stable value of 620-/+47 Omega.cm(2), the mannitol efflux rate was lower than 0.3%.h(-1).cm(-2), and the alkaline phosphatase activity in the apical side was significantly higher than that in the basolateral side.</p><p><b>CONCLUSION</b>The established Caco-2 cell model shows similar morphology to intestinal epithelial cells with formation of polarity, and can be used as an in vitro model for absorption studies.</p>

Epidemiological survey of the association between non alcoholic fatty liver disease and metabolic syndrome in civil servants of Chongqing city / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the association between non alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MS).</p><p><b>METHODS</b>A cross-sectional multiple-stage stratified survey was performed. A total of 2190 civil servants of Chongqing city were invited to participate in the survey covering physical examination, serum biochemistry-profile and ultrasonographic examination of liver.</p><p><b>RESULTS</b>Of 2176 valid questionnaires, altogether 455 cases were diagnosed as NAFLD and 231 individuals were diagnosed as MS. The prevalence of obesity, hyperglycemia, blood lipid disturbance, primary hypertension, NAFLD and MS was 38.3%, 5.5%, 31.7%, 29.9%, 20.9% and 10.6% respectively, which was increased along with aging (chi2 = 31.775, P = 0.000; chi2 = 25.985, P = 0.000; chi2 = 44.818, P = 0.000; chi2 =149.802, P = 0.000; chi2 = 61.302, P = 0.000; chi2 = 43.508, P = 0.000 a partly). The prevalence of obesity, hyperglycemia, dyslipidemia, primary hypertension, metabolic syndrome was significantly higher than those in control group (chi2 = 384.554, P = 0.000; chi2 = 25.597, P = 0.000; chi2 = 370.849, P = 0.000; chi2 = 40.252, P = 0.000; chi2 = 215.077, P = 0.000 separately), and the level of body mass index (BMI), fasting plasma glucose (FPG), triglyceride (TG), systolic blood pressure (SBP), diastolic blood pressure (DBP) in NAFLD group was remarkably higher than those in control group (t = 26.308, P = 0.000; t = 6.055, P = 0.000; t = 15.980, P = 0.000; t = 10.550, P = 0.000; t = 13.628, P = 0.000 respectively), while the level of high-density lipoprotein cholesterol (HDL-C) was on the opposite (t = 20.067, P = 0.000). Compared with the control group, odds risk for NAFLD was 22.82 folds (95% CI: 12.64-41.19) in obesity, 20.97 folds (95% CI: 11.21-39.24) in hyperglycemia, 24.40 folds (95% CI: 13.51-44.07) in dyslipidemia, 15.73 folds (95% CI: 8.66-28.60) in primary hypertension, while the risk for NAFLD was the highest in MS (OR = 31.06, 95% CI: 17.12-56.35). There were simple or multiple metabolic disorders in 455 individuals diagnosed as NAFLD, and 21 case (4.6%) with obesity, hyperglycemia, dyslipidemia and primary hypertension.</p><p><b>CONCLUSION</b>NAFLD is closely related with MS, which may be considered as a feature of MS.</p>

Effects of chromium nanoparticle dosage on growth, body composition, serum hormones and tissue chromium in Sprague-Dawley rats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

This 6-week study was conducted to evaluate the effects of seven different levels of dietary chromium (Cr) (0, 75, 150, 300, 450, 600, and 1 200 ppb Cr) in the form of Cr nanoparticle (CrNano) on growth, body composition, serum hormones and tissue Cr in Sprague-Dawley (SD) rats. Seventy male SD rats (average initial body weight of (83.2+/-4.4) g) were randomly assigned to seven dietary treatments (n=10). At the end of the trial, body composition was assessed via dual energy X-ray absorptiometry (DEXA). All rats were then sacrificed to collect samples of blood, organs and tissues for determination of serum hormones and tissue Cr contents. The results indicated that lean body mass was significantly increased (P<0.05) due to the addition of 300 and 450 ppb Cr from CrNano. Supplementation of 150, 300, 450, and 600 ppb Cr decreased (P<0.05) percent body fat significantly. Average daily gain was increased (P<0.05) by addition of 75, 150, and 300 ppb Cr and feed efficiency was increased (P<0.05) by supplementation of 75, 300, and 450 ppb Cr. Addition of 300 and 450 ppb Cr decreased (P<0.05) the insulin level in serum greatly. Cr contents in liver and kidney were greatly increased (P<0.05) by the addition of Cr as CrNano in the dosage of from 150 ppb to 1 200 ppb. In addition, Supplementation of 300, 450, and 600 ppb Cr significantly increased (P<0.05) Cr content in the hind leg muscle. These results suggest that supplemental CrNano has beneficial effects on growth performance and body composition, and increases tissue Cr concentration in selected muscles.