Analysis of a pedigree affected with hereditary coagulation factor XII deficiency due to a homozygous 252delAsn deletion of F12 gene / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical phenotype and genetic basis of a consanguineous pedigree affected with hereditary coagulation factor XII (FXII) deficiency.@*METHODS@#Following extraction of genomic DNA, all exons and flanking regions of F12 gene were subjected to PCR amplification and Sanger sequencing. ClustalX-2.1-win and MutationTaster software was used to analyze the conservation and impact of the variants on protein function.@*RESULTS@#DNA sequencing showed that the proband carried a homozygous g.6753-6755delACA deletion (p.252delAsn) in exon 9 of the F12 gene, for which her father, mother and brother were heterozygous carriers. The same deletion was not found in her sister.@*CONCLUSION@#The homozygous p.252delAsn deletion probably underlies the hereditary FXII deficiency in this pedigree.

Analysis of a consanguineous pedigree affected with hereditary coagulation factor XII deficiency caused by homozygous Gly341Arg mutation / 中华医学遗传学杂志

OBJECTIVE To analyze the laboratory phenotype and FXII gene mutation in a consanguineous Chinese pedigree affected with factor XII (FXII) deficiency. METHODS Activated partial thromboplastin time (APTT), FXII activity (FXII:C) and FXII antigen (FXII:Ag) of the proband and her family members (10 individuals from 3 generations) were determined. Sanger sequencing was used to detect potential mutation within the 14 exons, their flanking regions and 5',3'-untranslated regions of the FXII gene. Suspected mutations were verified by backward sequencing. Conservation of the amino acids were analyzed with ClustalX-2.1-win. Four online bioinformatics software (PolyPhen-2, PROVEAN, SIFT and MutationTaster) were used to assess the impact of the mutations on the protein function. RESULTS The APTT of the proband and her elder brother have prolonged to 61.6 s and 68.6 s,and their FXII:C and FXII:Ag have decreased to 12%, 10% and 11%, 10%, respectively. The APTT of the paternal grandmother, maternal grandmother, father, mother, elder paternal aunt and elder maternal aunt were all normal, but their FXII:C and FXII:Ag have reduced to half of the normal value. Gene sequencing found that the proband and her elder brother have both carried a homozygous missense c.1078G>A (p.Gly341Arg) mutation in exon 10 of the FXII gene, for which the paternal grandmother, maternal grandmother, father, mother, elder paternal aunt and elder maternal aunt were heterozygous. Bioinformatic analysis suggested that the Gly341 is highly conserved, while p.Gly341Arg is a harmful mutation which may cause disease by affecting the function of FXII protein. CONCLUSION Homozygous p.Gly341Arg mutation, caused by consanguineous marriage, probably underlies the congenital FXII deficiency in this pedigree.

Analysis of the influential factors of late diagnosis among newly identified HIV/AIDS cases in Anhui Province, 2011-2015 / 中华预防医学杂志

Objective@#To analyze the base situation and influential factors of late diagnosis among newly identified HIV/AIDS cases in Anhui Province from 2011 to 2015.@*Methods@#Database information of the newly identified HIV/AIDS cases in Anhui Province from 2011 to 2015 were downloaded from the National HIV/AIDS Comprehensive Information System of China's disease prevention and control information system. To analyze the data including basic information, sample source, route of HIV transmission, population mobility, venereal disease, death and first CD4 count; and the number of 7 073 cases were classified according to late diagnosis and non-late diagnosis criteria. The Chi-square test and logistic regression analysis were used to analyze the influential factors of HIV late diagnosis.@*Results@#A total of 7 073 newly identified HIV/AIDS cases were analyzed, and the mean age was (38.5±15.0) years. The proportion of late diagnosis in all counted cases was 41.7% (2 949/7 073); from 2011 to 2015, the proportions of late diagnosis were 59.7% (485/812), 46.5% (531/1 141), 42.7% (587/1 376), 36.1% (609/1 686), and 35.8% (737/2 058), respectively. Compared with the 0 to 19 years group, the 40 to 59 years group and over 60 years old group have higher risk of late diagnosis (OR=2.68, 95%CI: 1.94-3.71; OR=2.18, 95%CI: 1.53-3.10, respectively). Compared with the high education group, the illiterate and primary school education group have higher risk of late diagnosis (OR=1.74, 95%CI: 1.36-2.22; OR=1.64, 95%CI: 1.34-2.01, respectively). Compared with other sample sources, medical institutions have higher risk of late diagnosis (OR=2.64, 95%CI: 2.28-3.05). Compared with migrant population, the resident population have higher risk of late diagnosis (OR=1.80, 95%CI: 1.53-2.11).@*Conclusion@#The proportion of late diagnosis among newly identified HIV/AIDS cases in Anhui province was relatively high from 2011 to 2015. The main risk factors of late diagnosis included cases reported by medical institutions, resident population, over 40 years old age group and low education level.

Analysis of a pedigree affected with congenital hypofibrinogenemia due to heterozygous Ser313Ile mutation of fibrinogen γ chain gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the genetic basis for a Chinese pedigree affected with congenital hypofibrinogenamia.</p><p><b>METHODS</b>Peripheral blood samples were collected from 9 members from the pedigree. Routine coagulation tests including activated partial thromboplastin time (APTT), thrombin time (TT), the prothrombin time (PT) were carried out. The activity of fibrinogen (Fg: C) was measured using Clauss method, and fibrinogen antigen (Fg: Ag) was measured with immunoturbidimetry. All exons and exon-intron boundaries of the fibrinogen Aα, Bβ and γ chain genes were amplified using PCR, which was followed by direct sequencing. Suspected mutation was confirmed by reverse sequencing. The mutant fibrinogen was analyzed with Swiss-PdbViewer.</p><p><b>RESULTS</b>The proband showed prolonged APTT, PT and TT. Her functional fibrinogen (Fg: C) and antigen fibrinogen (Fg: Ag) levels were reduced to 0.69 g/L and 0.72 g/L, respectively. Her mother and grandmother also had a low levels of fibrinogen, which were 0.99 g/L and 0.83 g/L for Fg: C, 1.02 g/L and 0.87 g/L for Fg: Ag, respectively. The results of other members from the pedigree were all within the normal range. Genetic analysis reveled a heterozygous G>T mutation at nucleotide 7590 in exon 8 of γ gene in the proband, which was predicted to be a novel Ser313Ile mutation. The mutation was also found in her mother and grandmother. Model analysis showed that the Ser313Ile mutation disturbed the hydrogen bonds between Ser313, Asn319 and Asp320. Moreover, the mutation also altered the mutual electrostatic force and affected the folding and instability of the mutant fibrinogen.</p><p><b>CONCLUSION</b>The heterozygous Ser313Ile mutation probably underlies the hypofibrinogenemia in this pedigree.</p>

Association of polymorphisms of NAPE-PLD and FAAH genes with schizophrenia in Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of polymorphisms of N-acyl-phosphatidylethanolamine-phospholipase D (DAPE-PLD) and fatty acid amide hydrolase (FAAH) genes, as well as their interaction, with schizophrenia.</p><p><b>METHODS</b>Polymorphisms of NAPE-PLD rs12540583 and FAAH rs324420, rs2295633, and rs6429600 were determined with PCR - restriction fragment length polymorphism assay and Sanger sequencing. The genotypes of 345 subjects of Han Chinese origin diagnosed with schizophrenia and a 403 controls were compared. The results were analyzed with SPSS 17.0, and the interaction of the two genes was analyzed using a multifactor dimensionality reduction (MDR) method.</p><p><b>RESULTS</b>The frequency of NAPE-PLD rs12540583 polymorphism was significantly different between the two groups under both dominant and additive models (χ2=17.18 vs. χ2=18.94, P<0.0125). The frequencies of AC genotype and C allele of the patient group at rs12540583 were higher than those of the controls, and the interaction of NAPE-PLD and FAAH was associated with schizophrenia. A four-loci model (rs12540583, rs324420, rs2295633 and rs6429600) can best model the interaction between NAPE-PLD and FAAH.</p><p><b>CONCLUSION</b>The AC genotype and C allele of NAPE-PLD rs12540583 locus are risk factors for schizophrenia, and the interaction between NAPE-PLD rs12540583 and FAAH rs324420, rs2295633 and rs6429600 is associated with schizophrenia.</p>

Analysis of inherited dysfibrinogenemia pedigree associated with a heterozygous and deletion mutantion in the FGG gene / 中华检验医学杂志

Objective To analyze the phenotype and genotype of inherited dysfibrinogenemia pedigree associated with a novel heterozygous and deletion mutation in the FGG gene,and to investigate its molecular mechanism.Methods The clinical data were collected from the proband found at our hospital and her family members in April 2016.The activity plasma fibrinogen(Fg:C), activated partial thromboplastin time(APTT),prothrombin time(PT), thrombin time(TT)were detected by coagulation method and the antigen plasma fibrinogen(Fg:Ag), D-Dimer(D-D), fibrinogen degradation products (FDPs)were analyzed by immunoturbidimetry method.All of the exons and exon-intron boundaries of the genes of FGA, FGB and FGG with the fibrinogen(Fg)were amplified by PCR and followed by direct sequencing.And further verification were performed by cloning sequence and non-denatured polyacrylamide gel electrophoresis and silver staining.The conservatism of mutated gene locus were analyzed by ClustalX-2.1-win.The change of the protein spatial structure and the intermolecular forces with mutation were analyzed by Pymol.Results The Fg:C of the proband was significantly reduced(0.30 g/L)and the Fg:Ag of the proband was normal(2.00 g/L).Their Fg:C were both significantly reduced and the Fg:Ag were both normal(0.42 g/L,2.09 g/L & 0.47 g/L,2.42 g/L, respectively), these were found in her mother and grandma.Genetic analysis revealed a novel heterozygous and deletion mutation with c.944 _c.952 delCCTTTGATG in exon 8 of FGG gene in the proband,predicting a heterozygous 289_291delAla,Phe,Asp mutation.The same mutations were carried by her mother and grandma, but her father and grandpa were normal.Homology analysis indicated that the Ala 289,Phe290 and Asp291 were maintained highly conservative in homogenous species.Protein model analysis found that the original hydrogen bonds were disappeared when the deletion mutation happened with the Ala 289,Phe290 and Asp291.Conclusion The heterozygous and deletion mutation with 289_291delAla,Phe,Asp in the γchain of fibrinogen were identified that could cause the rearrangement of the Fg molecular space structure and the reduction of the structure stability,so the mutation probably underly the dysfibrinogenemia in this pedigree.(Chin J Lab Med,2018, 41:305-311)

Homozygous mutation Gly542Ser in one pedigree with congenital factor Ⅻ deficiency / 中华检验医学杂志

Objective To analyze the mutations of F12 gene in one pedigree with congenital factor FⅫ(FⅫ)deficiency, and investigate the molecular mechanisms of FⅫ deficiency.Methods Pedigree investigation.In February 2015,a patient with hereditary FⅫdeficiency was admitted to the Third Clinical College of Wenzhou Medical University.Activated partial thromboplastin time(APTT), prothrombin time (PT),FⅫactivity(FⅫ:C),FⅫ antigen(FⅫ:Ag)and other coagulant parameters were tested in the proband and his family members.5′and 3′UTR, all exons and their exon-intron boundaries of F12 gene were analyzed by direct sequencing.The detected mutations were confirmed by reverse sequencing.The conserved amino acids were analyzed by ClustalX-2.1-win software, and four bioinformatics softwares (PolyPhen-2,PROVEAN,SIFT and MutationTaster)were also used to analyze the effect of mutations on protein function.Results The proband and her younger brother showed a markedly prolonged APTT which were 116.4 s and 101.3 s, while her father had slightly prolonged APTT, and other family members were normal.The FⅫ:C and FⅫ:Ag of family members were also decreased(the proband,2.0% and 1.0%;her younger brother,2.0% and 1.0%; her father,18.0% and 13.0%).The phenotype of all members was consistent with cross-reactive material(CRM)negative.Nucleotide sequencing analysis showed that the proband and her younger brother had missense mutations in the F 12 gene, including one homozygous mutation c.1681G>A(p.Gly542Ser)and a commonly reported single nucleotide polymorphism site within the promoter region of the F12 gene(46T/T).Sequencing results from the proband's parents and son demonstrated them as carriers of a heterozygous missense mutation.The proband's husband was normal and with 46C/C in the promoter region.The ClustalX-2.1-win results indicated that the Gly542 was highly conserved among the homologousspecies.The predicting outcomes of the four bioinformatics softwares were the same,the PolyPhen-2(score 1.000)and PROVEAN(score -4.975)both declared p.Gly542Ser was a harmful mutation.The SIFT(score 0.00)and the MutationTaster(score 0.999)manifested the mutation could affect the protein funtion.Conclusions c.1681G>A(p.Gly542Ser)in exon 14 and 46T/T were related with the significant decrease of the FⅫlevel of this pedigree of hereditary FⅫ deficiency.

Lipopolysaccharides-stimulated macrophage products enhance Withaferin A-induced apoptosis via activation of caspases and inhibition of NF-κB pathway in human cancer cells.

Macrophages, as a major cellular component in tumor microenvironment, play an important role in tumor progression. However, their roles in modulation of cytotoxic chemotherapy are still not fully understood. Here, we investigated the influence of Lipoplysaccharides (LPS)-stimulated macrophage products (LSMP) on Withaferin A (WA), a natural compound that derived from the medicinal plant Withania somnifera, as an antitumor agent in human breast cancer cells MDA-MB-231 and prostate cancer cells PC-3. Our results revealed that LSMP may enhance WA-induced apoptosis in both cell lines, the underlying mechanisms of which are closely associated with activation of caspase-8, -9 and -3, cleavage of poly ADP-ribose polymerase (PARP), as well as specifically inhibiting the translocation of nuclear factor-κB (NF-κB) and down-regulation of anti-apoptotic proteins X-linked inhibitor of apoptosis protein (XIAP) and inhibitor of apoptosis protein (cIAP1/2). These findings demonstrate that macrophages in tumor microenvironment can modulate tumor responses to chemotoxic agents, providing an effective strategy that targets macrophages to enhance the antitumor efficacy of cytotoxic chemotherapy.

Effects of Jiuxieling Granules on Expressions of MyD88 and IRAK1 in Ulcerative Colitis Rats with Spleen-Kidney Yang Deficiency / 中国中医药信息杂志

Objective To observe the effects of Jiuxieling Granules on the expressions of MyD88 and IRAK1 in ulcerative colitis model rats with spleen-kidney yang deficiency; To discuss its mechanism of action. Methods Animal models were established by compound methods. 90 Wistar rats were randomly divided into blank group, model group, positive medicine group, and Jiuxieling Granules high-, medium-, and low-dose groups. Each administration group was given relevant medicine for gavage. RT-qPCR, SP immunohistochemistry and Western blot were used to detect mRNA and proteins of MyD88 and IRAK1 in colon tissues. Results Compared with the blank group, mRNA and proteins of MyD88 and IRAK1 in the model group increased (P<0.01). Compared with the model group, mRNA and proteins of MyD88 and IRAK1 in each administration group decreased (P<0.01), especially in Jiuxieling Granules high-dose group. Conclusion Jiuxieling Granules can reduce the expressions of MyD88 and IRAK1, and then influence the transmission of MyD88 signaling pathway and block the release of downstream inflammatory factors to achieve the result of treating ulcerative colitis with spleen-kidney yang deficiency.

Effects of Modified Danggui Beimu Kushen Pills on Toll-like Receptor Pathway Markers on MFC Gastric Cancer Mice / 中国中医药信息杂志

Objective To study the effects of modified Danggui Beimu Kushen Pills on the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissues on MFC gastric cancer bearing mice; To discuss relevant mechanism of action. Methods MFC gastric cancer bearing mice were employed to perform anti-tumor experiment in vivo in this study. A total of eligible 48 mice were randomly divided into model group, DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups, modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. The treatment was conducted once a day, and lasted for 14 continuous days. After the last administration of gavage orally treatment, all mice were anaesthetized and killed by cervical dislocation method to obtain tumor tissue completely for further HE staining measure and detection of TLR2, TLR4, TLR6, TRAF6 and MyD88 in tumor tissue with the method of RT-qPCR and immunohistochemistry. Meanwhile, the tumor growth was observed and the general conditions of mice were recorded. Results The model group was rich in tumor cells; the sizes of cells were different; the volume was large; the nucleus was deeply stained and the heterotypic shape was obvious, and the small focal necrosis was seen. The number of tumor cells in each administration group was reduced; the arrangement was loose; the cell volume was significantly reduced, and the nuclear pyknosis was reduced. Cell necrosis significantly increased; the number of interstitial blood vessels decreased; collagen fibers increased, especially in modified Danggui Beimu Kushen Pills high-dose and low-dose combined with DDP groups. Compared with the model group, the expressions of TLR2, TLR4, TLR6, TRAF6 and MyD88 mRNA and protein decreased in each administration group. TLR2, TLR4, TLR6, TRAF6 and MyD88 were lighter and weakly positive expressed in modified Danggui Beimu Kushen Pillshigh-dose and low-dose combined with DDP groups, the protein changes were more obvious Compared with DDP positive control group, modified Danggui Beimu Kushen Pills high-dose and low-dose groups. Conclusion Modified Danggui Beimu Kushen Pills can down-regulate TLR2, TLR4, TLR6, TRAF6 and MyD88 expression of tumor tissue in MFC gastric cancer bearing mice at both mRNA and protein levels to play anti-tumor pharmacology action.

Clinical curative effect of balloon occlusion of distal abdominal aorta in cesarean section for patients with pernicious placenta previa and placenta accreta / 中国介入影像与治疗学

Objective To analyze the clinical efficacy of balloon occlusion of distal abdominal aorta for patients with pernicious placenta previa and placenta accreta.Methods Data of 72 patients with pernicious placenta previa and placenta accreta were retrospectively analyzed.There were 53 cases (occlusion group) reserved balloon occlusion in abdominal aorta before cesarean section,which can temporarily blocked abdominal aortic blood flow during operation.The other 19 cases (non-occlusion group) underwent cesarean section without balloon occlusion of abdominal aorta.The intraoperative,post operative situations and the birth state of newborn of the two groups were compared.Results The bleeding,blood transfusion and hysterectomy rate during the operation in occlusion group were less than those in non-occlusion group (all P＜ 0.05).Differences of the rate of postoperative transferring to intensive care unit (ICU) and the time in ICU were statistically significant between two groups (both P ＜0.05).No statistical difference of operation time,postoperative total hospital stay time and the rate of postoperative infection was found between two groups (both P＞0.05).There was no statistical difference of newborns weight and Apgar scores (5 min and 10 min after birth) between two groups (all P＞0.05).Conclusion The balloon occlusion of distal abdominal aorta in cesarean section for patients with pernicious placenta previa and placenta accreta is safe and feasible,which can effectively reduce the intraoperative bleeding,the blood transfusion and the risk of hysterectomy.

Clinical features and coronary artery lesions in middle-aged and young patients of coronary heart disease complicated with diabetes mellitus / 中国医师进修杂志

Objective To discuss the clinical features and coronary artery lesions in middle-aged and young patients of coronary heart disease complicated with diabetes mellitus. Methods The clinical data of 386 middle-aged and young patients (aged from 28 to 59 years) of coronary heart disease were retrospectively analyzed. Among them, 176 cases were complicated with diabetes mellitus (diabetes group) and 210 cases didn′t have diabetes mellitus (non diabetes group). The general information, echocardiography and coronary angiography results between 2 groups were compared. Results The female ratio, incidences of hypertension and body mass index (BMI) in diabetes group were significantly higher than those in non diabetes group:28.4%(50/176) vs. 6.7%(14/210), 63.6%(112/176) vs. 43.8%(92/210) and (26.5 ± 2.5) kg/m2 vs. (23.8 ± 1.7) kg/m2, and there were statistical differences (P0.05). The total cholesterol (TC) and triglyceride (TG) in diabetes group were significantly higher than those in non diabetes group:(4.96 ± 1.32) mmol/L vs. (4.67 ± 1.12) mmol/L and (2.77 ± 2.01) mmol/L vs. (2.09 ± 1.60) mmol/L, the high-density lipoprotein cholesterol (HDL-C) was significantly lower than that in non diabetes group:(0.92 ± 0.30) mmol/L vs. (1.10 ± 0.32) mmol/L, and there were statistical differences (P0.05). There were no significant differences in the left atrial diameter, right ventricular diameter, left ventricular end diastolic diameter, ejection fraction, aortic root inside diameter and incidence of weakened ventricular wall motion between 2 groups (P>0.05). The incidence of 3 branch coronary artery lesions in diabetes group was significantly higher than that in non diabetes group:59.1%(104/176) vs. 37.6%(79/210), and there was statistical difference (P0.05). Conclusions In middle-aged and young patients of coronary heart disease complicated with diabetes mellitus, the female ratio is higher and hypertension, obesity and hyperlipidemia is more common. The lesions of coronary artery are more serious and diffuse. Three branch coronary artery lesions are the main ones, and the left main branch and the circumflex branch lesion is common.

Phenotypic and genetic analysis of two pedigrees affected with hereditary antithrombin deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the phenotype, genotype and molecular mechanism for two pedigrees affected with hereditary antithrombin (AT) deficiency.</p><p><b>METHODS</b>Clinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, antithrombin activity (AT:A) and specific antigen (AT:Ag), protein C activity, as well as protein S activity. To detect potential mutations in the probands, all exons, exon-intron boundaries and the 3', 5' untranslated regions were amplified by PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and silver staining. The effect of mutations on the AT protein was analyzed with bioinformatics software.</p><p><b>RESULTS</b>The AT:Ag of pedigree 1 was normal, but its AT:A has reduced to 30%. A heterozygous c.235C>T mutation in exon 2 causing p.Arg47Cys, in addition with two single nucleotide polymorphisms (c.981G>A, c.1011G>A) in exon 5 were identified in the patient. His four children, except for the elder daughter, were heterozygous for the mutations. The plasma levels of AT:A and AT:Ag in proband 2 have decreased to 39% and 103 mg/L, respectively. A heterozygous deletion (g.5890-5892delCTT) leading to loss of p.Phe121 was also detected in his father. Bioinformatic analysis suggested that the missense mutation Arg47Cys can affect the functions of AT protein. Meanwhile, lacking of Phe121 will result in loss of hydrogen bonds with Ala124, Lys125 and the cation π interactions with Lys125, Arg47, which may jepordize the stability of the protein.</p><p><b>CONCLUSION</b>The proband 1 had type II AT deficiency, while proband 2 had type I AT deficiency. The p.Arg47Cys and g.5890-5892delCTT mutations of the AT gene are significantly correlated with the levels of AT in the two probands, respectively.</p>

Homozygous missense mutation p.Val298Met of F10 gene causing hereditary coagulation factor X deficiency in a Chinese pedigree / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential mutation underlying coagulation factor X (FX) deficiency in a consanguineous Chinese pedigree.</p><p><b>METHODS</b>Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FX activity (FX:C) and other coagulant parameters were determined with a one-stage clotting assay. The FX antigen (FX:Ag) was determined with an ELISA assay. All coding exons and exon-intron boundaries of the F10 gene were amplified with PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and analyzed with CLC Genomics Workbench 7.5 software.</p><p><b>RESULTS</b>The PT and APTT in the proband were prolonged to 67.2 s and 102.9 s, respectively. Further study showed that her FX:C and FX:Ag were reduced by 1% and 8%, respectively. The PT of her father, mother, and little brother were slightly prolonged to 14.5 s, 14.4 s and 14.4 s, respectively. The FX:C and FX:Ag in her father, mother and little brother were all slightly reduced. Genetic analysis of the proband has revealed a homozygous G>A change at nucleotide 27881 in exon 8 of the F10 gene, which predicted a p.Val298Met substitution. The proband's father, mother, and little brother were all heterozygous for the p.Val298Met mutation. The proband has inherited the homozygous mutation from her parents by consanguineous marriage. Other family members were all normal. Bioinformatics analysis has indicated that this mutation may result in changes in the secondary structure of the FX protein.</p><p><b>CONCLUSION</b>A homozygous mutation g.27881G>A(p.Val298Met) of the F10 gene has been identified, which probably accounts for the low FX concentrations in this pedigree.</p>

Effects of RASI on expression of airway MUC5 AC and related inflammatory factors in asthmatic mice with Yin deficiency syndrome / 中国免疫学杂志

Objective:To study the effects of Radix Angelicae Sinensis ( RASI) on expression of airway MUC5AC and related inflammatory factors in asthmatic mice with Yin deficiency syndrome.Methods:Injecting ovalbumin ( OVA) to sensitize,inhaling OVA to stimulate,using Thyroxin during late stimulation,the asthmatic mouse with Yin deficiency syndrome was established and evaluated through asthmatic behaviors, lung histopathology, active factors ( IL-13, TNF-αand MUC5AC ) in broncho-alveolar lavage fluid (BALF), and MUC5AC expression in lung tissue.Results: 2,4,8 g/kg RASI could reduce asthmatic behaviors score, relieve pathological changes of lung tissue,reduce the contents of IL-13,TNF-αand MUC5AC in BALF,and depress MUC5AC expression in lung tissue ( P<0.05,0.01).In addition,there was a certain synergy between RASI and dexamethasone ( DXM) on depressing the ex-pression of IL-13 and MUC5AC (P<0.05).Conclusion:RASI has certain anti-asthma effect and one of mechanisms is to regulate the MUC5AC expression through inhibit IL-13 and TNF-α.On the expression of IL-13 and MUC5AC,the compatibility of RASI with glu-cocorticoid has some synergy effect.

Establishment and Evaluation of BALB/c Mouse Model of Anaphylactic Asthma with ;Yin-deficiency Syndrome / 中国中医药信息杂志

Objective To establish and evaluate a BALB/c mouse model of anaphylactic asthma with yin-deficiency syndrome. Methods Ovalbumin (OVA) was injected to sensitize and was inhaled to stimulate to replicate the BALB/c mouse model of anaphylactic asthma. Thyroxin was used for gavage during late stimulation to replicate the BALB/c mouse model of asthma with yin-deficiency syndrome. On the basis of monitoring food and water intake and body weight, asthma related indexes, such as asthmatic behaviors, respiratory function, lung histopathology, were detected, and yin-deficiency syndrome related indexes, such as cAMP and cGMP, and pulmonary fluid clearance were examined. Results The BALB/c mouse model of asthma with yin-deficiency syndrome showed obvious asthma symptoms; comprehensive scores of asthmatic behaviors increased significantly;inhaling peak flow, exsufflation peak flow, and tidal volume decreased significantly; lung tissue histopathology showed obvious inflammatory response. Meanwhile, food and water intake of BALB/c mouse model of asthma with yin-deficiency syndrome increased significantly;body weight increased slowly;wet/dry ratio of lung tissue decreased;cAMP and cAMP/cGMP increased, while cGMP decreased, which showed that the mice were in the condition of yin-deficiency and yang-excess. Conclusion Combining thyroxin at the basis of sensitivity induced by OVA is a good pattern to successfully replicate the BALB/c mouse model of anaphylactic asthma with yin-deficiency syndrome.

Phenotype and genotype analysis of in herited dysplasminogenemia caused by plasminogen Ala601Thr mutation / 中华检验医学杂志

Objective To detectthe phenotype and gene mutations underlying aninherited dysplasminogenemia pedigree and search the virulence gene.Methods The peripheral venous blood samples of the proband and his family members (fourteen subjects of three generations in total) were collected,and their prothrombin time(PT),activated partial thromboplastin time(APTF),thrombin time(TT),fibrinogen (FIB),fibrinogen degradation products (FDP),D-dimmer (D-D)weretested on a STAGO analyzer,the plasminogen activity (PLG:A) and plasminogen antigen (PLG:Ag) were analyzedby thechromogenic substrate assay and rocket immunoelectrophoresis,respectively.All 19 exons,5' and 3' untranslated regions of PLGwere amplified with PCR.Direct DNA sequencing was used to analyze the amplified products,which wereconfirmed by backward sequencing.Three bioinformatics online softwares (SIFT,PolyPhen-2 andMutationTaster) were used to forecast the possible impact of the mutations on the protein function.At last,themodel analysis of mutate site was taken on a Swiss-Pdb Viewer software.Results The PLG:Avalue of theproband and other 6 family members were decreased to the half,while the PLG:Ag was normal.The D-Dand FDP value of the proband,his grandma and father were slightly higher.DNA sequencing has revealedthat the proband and the other 6 members of this family had the same mutation of g.38829G ＞ A in exon 15,leading to the missense mutationp.Ala601Thr.The results of bioinformatics softwares showed that themutation could affect the thePLGfunction.Protein model analysis indicated that the hydrophobic interaction force and hydrogen bond between the amino acids were changed,which might affect the stability of the PLG.In addition,all the members of this family take the heterozygous SNP of g.2501C ＞ A in the 5 ＇UTR.Conclusions The p.Ala601Thr found in the inherited dysplasminogenemia pedigree in the exon 15 was responsible for the reduced PLG:A of the family,the dysplasminogenemia and this mutation were both reported for the first time in China.

Advances in Study on Risk Factors Related to Bile Reflux / 胃肠病学

Bile reflux is not only related to diseases such as gastritis,esophagitis,pharyngitis,chorditis,bronchitis and pneumonia,but also related to residual gastric ulcer,residual gastric cancer,intestinal metaplasia,dysplasia and carcinogenesis. Risk factors related to bile reflux include various operation modes, various anastomosis methods, gallbladder stone, cholecystectomy and various non-operative factors such as age, gender, allergy, mental and psychological factors,congenital factors. This article reviewed the advances in study on risk factors related to bile reflux.

Preparation of a rat model of ulcerative colitis with spleen and kidney Yang deficiency / 中国实验动物学报

Objective To establish a rat model of ulcerative colitis ( UC) with spleen and kidney Yang deficien-cy.Methods The rat model of ulcerative colitis with spleen and kidney Yang deficiency was established by oral adminis-tration of Rhubarb decoction, intramuscular injection of hydrocortisone, and combined with ethanol enema of TNBS (2,4, 6-trinitrobenzene sulfonic acid).Sixty Wistar rats ( body weight 180 ±20 g, male:female=1∶1) were randomly divided into blank control group and groups of UC models with spleen kidney Yang deficiency for 7 days, 14 days and 21days.The serum levels of FT3, FT4, and T of the rats were detected by ELISA assay.Results Compared with the blank control group, the serum levels of FT3, FT4, and T in the groups of UC rat models with spleen kidney Yang deficiency for 7 days, 14 days and 21days were decreased to a different extent (P<0.05), especially, decreased dramatically in the model group for 21 days.Conclusions FT3, FT4, and T are sensitive indexes with spleen and kidney Yang deficiency.The detection of serum levels of FT3, FT4, and T can better verify the spleen and kidney Yang deficiency in the rats, and prove that the spleen and kidney Yang deficiency type UC animal model is successfully prepared.