Analysis of A Pedigree with Hereditary Coagulation Factor Ⅻ Deficiency Caused by Compound Heterozygous Mutations / 中国实验血液学杂志

OBJECTIVE@#To analysis clinical phenotype and potential genetic cause of a family affected with hereditary coagulation factor Ⅻ deficiency.@*METHODS@#The prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), D-Dimer (D-D), coagulation factor Ⅻ activity (FⅫ:C) and coagulation factor Ⅻ antigen (FⅫ:Ag) were determined for phenotype diagnosis of the proband and his family members(3 generations and 5 people). Targeted capture and whole exome sequencing were performed in peripheral blood sample of the proband. Possible disease-causing mutations of F12 gene were obtained and further confirmed by Sanger sequencing. The corresponding mutation sites of the family members were analyzed afterwards. The online bioinformatics software AutoPVS1 and Mutation Taster was used to predict the effects of mutation sites on protein function.@*RESULTS@#The APTT of the proband was significantly prolonged, reaching 180.9s. FⅫ:C and FⅫ:Ag of the proband was significantly reduced to 0.8% and 4.17%, respectively. The results of whole exome sequencing displayed that there were compound heterozygous mutations in F12 gene of the proband, including the c.1261G＞T heterozygous nonsense mutation in exon 11 (causing p.Glu421*) and the c.251dupG heterozygous frameshift mutation in exon 4 (causing p.Trp85Metfs*53). Both mutations are loss of function mutations with very strong pathogenicity, leading to premature termination of the protein. AutoPVS1 and Mutation Taster software predicted both mutations as pathogenic mutations. The results of Sanger sequencing revealed that c.1261G＞T heterozygous mutation of the proband was inherited from his mother, for which his brother and his daughter were c.1261G＞T heterozygous carriers. Genotype-phenotype cosegregation was observed in this family.@*CONCLUSION@#The c.1261G＞T heterozygous nonsense mutation in exon 11 and the c.251dupG heterozygous frameshift mutation in exon 4 of the F12 gene probably account for coagulation factor Ⅻ deficiency in this family. This study reports two novel pathogenic F12 mutations for the first time worldwide.

Feng-liao-chang-wei-kang is synergistic with 5-fluorouracil in inhibiting proliferation of colorectal cancer

Objective: To explore the effect of Feng-liao-chang-wei-kang (FLCWK) on acute and chronic gastroenteritis, synergistic effect on the growth inhibitory effect with 5-fluorouracil (5-FU) on colorectal cancer and its underlying mechanisms. Methods: In the in vitro study, HT-29 cells were divided into 5-FU alone, FLCWK alone and coadministration groups. The MTT assay was used to analyze the proliferation of HT-29 cells at 24 h. Flow cytometry was used to observe the apoptosis, cycle of colorectal cancer HT-29 cells at 24 h. In the in vivo experiment, The subcutaneous transplantation tumor model of colorectal cancer HT-29-Luc was established with nude mice. All mice were randomly divided into 5-FU alone, FLCWK alone and coadministration groups according to body weight. During administration, the Interactive Video Information System small animal live imaging system was used to monitor the growth of subcutaneous transplantation in nude mice. The model of colitis-associated colorectal cancer (CACC) was established with BALB/c mice. BALB/c mice were randomly divided into the normal control group, the model control group, and the FLCWK group. At the end of the administration, the pathological status was detected by HE staining. Cell apoptosis of tumor tissue tumor and colon tissues were observed by TUNEL staining and TUNEL green fluorescence. The protein expression of Caspase 3, p-STAT3, Bcl-2, Bax and P-gp in tumor tissues tumor and colon tissues were tested by using immunohistochemical assay. Results: FLCWK and 5-FU coadministration suppressed HT-29 cell viability and induced S phase arrest and apoptosis compared to treatment with 5-FU alone. Furthermore, compared to treatment with 5-FU alone, coadministration of FLCWK and 5-FU obviously reduced tumor volume and weight and induced apoptosis through decreasing p-STAT3 and P-gp and increasing Caspase 3 protein expression in a murine xenograft tumor model. Moreover, the result revealed decreased number and size of tumors following FLCWK protective administration, downregulated p-STAT3 and Bcl-2 levels and upregulated Bax and Caspase 3 expression in mice with CACC. Conclusions: FLCWK has synergistic effects with 5-FU on colorectal cancer by suppressing the STAT3 pathway and downregulating P-gp expression. Furthermore, FLCWK administration suppresses CACC tumorigenesis by inhibiting the STAT3 pathway.

Simultaneous Determination of Furan and Vinyl Acetate in Vapor Phase of Mainstream Cigarette Smoke by GC-MS

ABSTRACT A simple and sensitive method for simultaneous determination of furan and vinyl acetate (VA) in vapor phase of mainstream cigarette smoke with cold trap and gas chromatography-mass spectrometry (GC-MS) was developed. A Cambridge filter pad (CFP) was placed in front of the impingers of smoking machine to remove the particle phase from cigarette smoke. Furan and VA in vapor phase of mainstream cigarette smoke were collected in two impingers connected in series by filled with methanol at -78°C. The solutions were added with deuterium-labeled furan-d4 and VA-d6 as internal standards and analyzed by GC-MS. The results showed that the calibration curves for furan and VA were linear (r2 > 0.9995) over the studied concentration range. The intra- and inter-day precision values for furan and VA were <7.07% and <9.62%, respectively. The extraction recoveries of furan and VA were in the range of 94.5-97.7% and 92.3-94.9%, respectively. Moreover, the limits of detection for furan and VA were 0.028 µg mL-1 and 1.3 ng mL-1, respectively. The validated method has been successfully applied to determine the emissions of furan and VA in the vapor phase of mainstream cigarette smoke under International Organization for Standardization (ISO) and Canadian Intense (CI) smoking regimen.

Synthesis and antifungal activities of N-1,3,4-thiadiazol-2-yl-4-oxo-thiochroman-2-yl-formamide derivatives / 药学学报

Thiochromanones and 1,3,4-thiadazoles as heterocyclic compounds have broad biological activities. In order to find novel compounds with antifungal bioactivity, substituted thiophenol and maleic anhydride were used to synthesize the intermediate 4-oxothiochromane-2-carboxylic acid. It was reacted with 2-amino-1,3,4-thiadiazol to get fourteen target compounds containing 1,3,4-thiadazole moiety. The structures of the obtained compounds were confirmed by 1H NMR, 13C NMR and HR-MS. All compounds were investigated for antifungal activity via microdilution broth method. The results showed that the target compounds 3a and 3c to Epidermophyton floccosum and Mucor racemosus exhibited better antifungal activity than the positive control fluconazole, in which the minimum inhibition concentration can reach 8 μg·mL-1 and 16 μg·mL-1. Compound 3e showed significant inhibitory activity to Helminthosporium maydis, Sclerotinia sclerotiorum and Botrytis cinerea compared with that of the positive control carbendazim. Compound 3b exhibited inhibitory activity to Helminthosporium maydis better than the positive control carbendazim.

Correlation between atypical respiratory pathogens infection and vitamin D level in children / 重庆医学

Objective To investigate the correlation between atypical respiratory pathogens infection and serum vitamin D(VitD) level in children.Methods Serum IgM antibody levels of 11 atypical respiratory pathogens were detected in 414 serum samples of child respiratory infection by using the respiratory tract 11-items detection reagent kits (indirect immunofluorescence assay),including respiratory syncytial virus(RSV),adenovirus(ADV),influenza A virus(FluA),influenza B virus(FluB),parainfluenzavirus(PFlu),Mycoplasma pneumonia(MP),Chlalmydia pneumoniae(CP),Coxsackie virus type B(CoxB),Coxsackie virus type A(CoxA) and legionella pneumophilia(LP).At the same time the electrochemiluminescence assay was used to measure serum VitD level.Results Among 414 samples,pathogen IgM was detected out in 214 samples (51.69%),the top three places of detection rates were FluB,FluA and Mp,their positive rates were 32.13%,23.19% and 13.77% respectively;in cases of positive IgM antibody,17.63% of children developed single infection,34.06% of children developed 2 kinds of pathogen or more mixed infection;there was no statistical difference in the VitD levels between the IgM antibody positive group (median 23.60 ng/mL,3.37-71.50 ng/mL) and the IgM antibody negtive group(median 23.95 ng/mL,3.00-81.70 ng/mL).The IgM antibody positive rate,single infection positive rate and mixed infection positive rate had no statistical difference between the VitD reduce group and the VitD normal group(P>0.05).The positive rate of FluB,FluA and MP IgM antibody had no statistical difference between the VitD reduce group and the VitD normal group(P>0.05).Atypical respiratory pathogens had no correlation with VitD(r=0.005,P=0.912).Conclusion Atypical respiratory pathogens infection may had no correlation with the VitD level reduce.

Analysis of clinical outcomes of simplified treatment of lumbar spondylolisthesis with adjacent segment degeneration by MAST Quadrant / 中国骨伤

<p><b>OBJECTIVE</b>To explore the method and clinical effect of MAST Quadrant for lumbar spondylolisthesis with adjacent segment degeneration.</p><p><b>METHODS</b>From April 2014 to January 2016, 36 cases of lumbar spondylolisthesis with adjacent segment degeneration were treated by MAST Quadrant(target nerve decompression and transforaminal lumbar interbody fusion or articulationes zygapophysiales fusion by unilateral fixation with MAST Quadrant). Twenty-three cases were degenerative lumbar spondylolisthesis and 13 cases were isthmic lumbar spondylolisthesis. According to Meyerding grade of spondylolisthesis, 16 cases were grade I, 17 cases were grade II, and 3 cases were grade III. Visual analogue score (VAS), Oswesty Disability Index (ODI) and JOA score were used to evaluate the clinical outcome.</p><p><b>RESULTS</b>The amount of intraoperative bleeding was 230 to 480 ml with an average of 340 ml and the amount of postoperative blood loss was 15 to 80 ml with an average of 43 ml. Operative time was 176 to 240 min with an average of 193 min; X-ray exposure time was 2 to 6 s with an average of 3.6 s. Two cases were complicated with dural tear without nerve injury during operation. Thirty cases were followed up from 12 to 17 months with an average of 15.2 months. VAS scores for preoperative, 5 days, 3 months after surgery were 7.6±1.7, 1.9±0.4, 0.8±0.4 respectively, and there was significant difference before and after operation(<0.05). The ODI scores for preoperative and 3 months after surgery were 35.9±1.2 and 3.7±0.7 respectively, and there was significant difference before and after operation(<0.05). JOA scores for preoperative, 5 days, 1 months, 3 months after surgery were 13.2±0.4, 24.4±0.4, 27.4±0.1, 27.9±0.5 respectively, and there was significant difference before and after operation(<0.05).</p><p><b>CONCLUSIONS</b>MAST Quadrant can be applied to treat lumbar spondylolisthesis with adjacent segment degeneration, and the minimally invasive sugical technique is a safe and effective method, with the advantage of simple operation, fast recovery.</p>

Synthesis, biological activity and molecular docking research of N-{(4-oxo-thiochroman-3-yl)phenyl-methyl}acetamide derivatives as α-glucosidase inhibitors / 药学学报

In order to develop potent antidiabetic agents that have inhibitory effect to α-glucosidase, twelve β-acetamido ketone derivatives such as N-{[(substituted-4-oxo-thiochroman-3-yl)phenyl]-methyl}acetamide are designed and synthesized through one-pot Dakin-West reaction. Their chemical structures are confirmed by 1H NMR, 13C NMR, IR and HR-MS. In vitro α-glucosidase inhibition assays of compounds 4a-4l were carried out using glucose oxidase method. The result indicated that most of them possess inhibitory activity in vitro. Compound 4k showed the most potent inhibitory activity with 87.3% inhibition of α-glucosidase at the concentration of 5.39 mmol·L-1. The structure-activity relationship of these β-acetamido ketone derivatives was discussed preliminarily. Moreover, the molecular docking method was used to study the interaction mode of compound 4k and α-glucosidase. Our results will be helpful for designing of α-glucosidase inhibitors in the future.

Synthesis, biological activity and molecular docking research of N-{(4-oxo-thiochroman-3-yl)phenyl-methyl}acetamide derivatives as α-glucosidase inhibitors / 药学学报

In order to develop potent antidiabetic agents that have inhibitory effect to a-glucosidase, twelve β-acetamido ketone derivatives such as N-{[(substituted-4-oxo-thiochroman-3-yl)phenyl]-methyl}acetamide are designed and synthesized through one-pot Dakin-West reaction. Their chemical structures are confirmed by 1H NMR, 13C NMR, IR and HR-MS. In vitro α-glucosidase inhibition assays of compounds 4a-41 were carried out using glucose oxidase method. The result indicated that most of them possess inhibitory activity in vitro. Compound 4k showed the most potent inhibitory activity with 87.3% inhibition of α-glucosidase at the concentration of 5.39 mmol x L(-1). The structure-activity relationship of these β-acetamido ketone derivatives was discussed preliminarily. Moreover, the molecular docking method was used to study the interaction mode of compound 4k and α-glucosidase. Our results will be helpful for designing of α-glucosidase inhibitors in the future.

Effect of FTY720 inhibiting corneal neovascularition induced by sphingosine 1-phosphate / 国际眼科杂志(Guoji Yanke Zazhi)

AlM: To explore the inhibiting effect of FTY720 on corneal neovascularization ( CNV) of rat.METHODS: MTT assay and cells scratch were adopted to observe hyperplasia of human umbilical vein endothelial cells ( HUVECs ) and cell migration induced by sphingosine-1-phosphate(S1P) after using FTY720 of different concentration. The effect of FTY720 on CNV induced by S1P in a rat corneal micropocket model was detected. 30SD rats were randomly divided into group A, group B and group C with 10 rats per group. S1P and 0μg, 5μg, and 20μg FTY720 controlled-released particles were implanted into the corneal stroma. The growth of CNV and having pathological examination on 12d after the operation was observed. Findings was analyzed by one-way ANOVA.RESULTS: 10, 102 , 103 , and 104 nmol/L FTY720 and HUVECs co-incubate 72h could inhibit cell proliferation (P < 0. 01 ), 24h after the function of 10, 100nmol/L FTY720, it could inhibit S1P-induced cell migration and the ability of restricting cell proliferation and cell migration was enhanced with increasing concentration of FTY720. On 12d, after rat corneal micropocket controlled-release particles was implanted into groups A, B, C, the CNV area were respectively 10. 05±1. 19, 6. 59±0. 95, 2. 70± 0.68mm2(F=145. 155, P<0. 01), group A and group B was statistically different and this was the same case between group B and group C (P<0. 01).CONCLUSlON:FTY720 can inhibit S1P-induced corneal neovascularization.

Effect of cyclosporine A on the Pl - 3 k pathway in proliferation of rat lens epithelial cells / 国际眼科杂志(Guoji Yanke Zazhi)

AlM:To investigate the effect of cyclosporine A ( CsA) on the phosphatidylinositol-3 kinase ( Pl-3k ) pathway during procession of proliferation in epithelial cells of rabbit lens, and provide treatment strategies for after cataract on the basis of experiment. METHODS: Sixty eyes of 30 healthy white rabbits were operated by lens cortex removal in cataract surgery, and 30 right eyes were divided in treatment group and the other 30 eyes were divided in control group. From the first postoperative day, the control group eyes were dropped with normal saline 6 times each day, and the treatment group eyes were dropped with 1% CsA 6 times each day. Six rabbits were selected randomly and killed on the day before dropping and 1, 2wk, 1 and 2mo of postoperative day respectively. The lens of those killed rabbits were removed by surgery. The strategies of immunohistochemistry and mount in situ hybridization were used to detect the content of proliferating cell nuclear antigen ( PCNA) , gene of phosphate and tension homology deleted on chromsome ten ( PTEN) mRNA, Ser473-R, respectively. RESULTS:The expression of PCNA and Ser473-R were both down-regulate after operation in treatment group and control group, and the PCNA levels were significantly lower among treatment group than those in control group on 1wk (0.690±0.035 vs 0.785±0.015, t=6.099, P CONCLUSlON:1% CsA could inhibit the proliferation of epithelial cells in lens of rabbits with after cataract through preventing Pl-3k pathway.