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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.
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Humanos , Afibrinogenemia/genética , Codón sin Sentido , Linaje , Fenotipo , Fibrinógeno/genética , GenotipoRESUMEN
Objective:To evaluate the role of miR-20a-5p in M1 microglia aggravating oxygen-glucose deprivation and restoration (OGD/R)-induced injury to neurons and the relationship with mitofusin2 (MFN2).Methods:The well-growing BV2 microglia (M0 type) were polarized into M1 phenotype by lipopolysaccharide (100 ng/ml) and IFN-γ (20 ng/ml) and identified by quantitative real-time polymerase chain reaction and immunofluorescence.The well-growing N2a cells were divided into 6 groups ( n=6 each) by the random number table method: control group (group C), OGD/R group, M0 microglia co-culture group (group M0), M1 microglia co-culture group (group M1), miR-20a-5p inhibitor transfection group (group I) and negative control group (group NC). The cells were routinely cultured in group C, and the cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply to develop the model of OGD/R injury in group OGD/R.The cells were subjected to OGD for 3 h and were co-cultured with M0 microglia for 24 h during restoration of oxygen-glucose supply in group M0.The cells were subjected to OGD for 3 h and were co-cultured with M1 microglia for 24 h during restoration of oxygen-glucose supply in group M1.In group I and group NC, cells were transfected with miR-20a-5p inhibitor and negative control miRNA into M1 microglia, respectively, and N2a cells were subjected to OGD for 3 h and co-cultured with M1 microglia for 24 h during restoration of oxygen-glucose supply.The cell viability was determined by cell counting kit-8 assay, amount of lactate dehydrogenase (LDH) released was determined, the expression of miR-20a-5p and MFN2 mRNA was detected by quantitative real-time polymerase chain reaction, and MFN2 expression was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the amount of LDH released was increased, and the expression of MFN2 protein and mRNA was down-regulated in the other five groups, miR-20a-5p expression was significantly up-regulated in OGD/R, M0 and M1 groups, and miR-20a-5p expression was significantly down-regulated in group I ( P<0.05). There were no significant differences in the cell viability, amount of LDH released, and expression of miR-20a-5p, MFN2 protein and mRNA between group OGD/R and group M0 ( P>0.05). Compared with group OGD/R and group M0, the cell viability was significantly decreased, the amount of LDH released was increased, and the expression of MFN2 protein and mRNA was down-regulated, and miR-20a-5p expression was up-regulated in group M1 ( P<0.05). Compared with group M1, the cell viability was significantly increased, the amount of LDH released was decreased, the expression of MFN2 protein and mRNA was up-regulated, and miR-20a-5p expression was down-regulated in group I ( P<0.05). Conclusions:The mechanism by which M1 microglia aggravates OGD/R-induced damage to N2a cells may be related to the up-regulation of miR-20a-5p expression in M1 microglia and the inhibition of MFN2 expression in N2a cells.
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Objective: To investigate the molecular pathogenesis and clinical features of unrelated 12 patients with inherited coagulation protein C (PC) deficiency in Chinese population. Methods: The PC activity (PC:A) and PC antigen (PC:Ag) were detected by chromogenic substrate and enzyme linked immunosorbent assay, respectively. The nine exons and flanking sequences of the protein C (PROC) gene were amplified by polymerase chain reaction with direct sequencing, and the suspected mutations were validated by reverse sequencing (clone sequencing for deletion mutations) . Results: The PC:A of the 12 probands decreased significantly, ranging from 18% to 55%, and the PC:Ag of the 10 probands decreased significantly. Eleven mutations were found, out of which four mutations [c.383G>A (p.Gly128Asp) , c.997G>A (p.Ala291Thr) , c.1318C>T (p.Arg398Cys) , and c.532G>C (p.Leu278Pro) ] were discovered for the first time. Six mutations were in the serine protease domain, four mutations were located in epidermal growth factor (EGF) -like domains, and one mutation was located in activation peptide. There were two deletion mutations (p.Met364Trp fsX15 and p.Lys192del) , and the rest were missense mutations. Mutations p.Phe181Val and p.Arg189Trp were identified in three unrelated families. All mutations may be inherited, and consanguineous marriages were reported in two families. Among the probands, nine cases had venous thrombosis, two cases had poor pregnancy manifestations, and one case had purpura. Conclusion: Patients with PC deficiency caused by PROC gene defects are prone to venous thrombosis, especially when there are other thrombotic factors present at the same time.
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Humanos , Mutación , Mutación Missense , Linaje , Fenotipo , Proteína C/genética , Deficiencia de Proteína C/genéticaRESUMEN
Objective:To evaluate the relationship between preoperative cerebrospinal fluid/serum albumin ratio (Q-alb) and postoperative delirium (POD) in patients undergoing neuraxial anesthesia.Methods:The patients, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, underwent total knee/hip replacement under combined spinal-epidural block in our hospital from January 2018 to December 2020, were collected.After admission to the operating room, venous blood and cerebrospinal fluid samples were collected for determination of cerebrospinal fluid albumin, β-amyloid (Aβ) 1-42, Aβ 1-40, total tau protein (t-Tau), phosphorylated tau protein (p-Tau) and serum albumin levels (by enzyme-linked immunosorbent assay) and for calculation of Q-alb.When Q-alb was more than 10.2, the patient was considered to have blood-brain barrier disruption.Mini-Mental State Examination scale was used to evaluate the cognitive level on 1 day before surgery. The development of POD was evaluated using Confusion Assessment Method Chinese Reversion and Memorial Delirium Assessment Scale at 1-7 days after surgery.The patients were divided into POD group (P group) and non-POD (NP group) according to whether POD occurred.The receiver operating characteristic (ROC) curve was used to analyze the accuracy of Q-alb in predicting POD. Results:There were 49 cases in each group.Compared with group NP, concentrations of Aβ 1-42 and Aβ 1-40 were significantly decreased, concentrations of t-Tau and p-Tau albumin were increased, the ratio of Q-alb and blood-brain barrier disruption was increased in group P ( P<0.05). Before and after adjusting for confounding factors, Q-alb, cerebrospinal fluid Aβ 1-42, Aβ 1-40, t-Tau and p-Tau levels were risk factors for POD ( P<0.05). There was a positive linear regression relationship between Q-alb and levels of t-Tau and p-Tauin cerebrospinal fluid (t-Tau: β=0.587, P<0.001; p-Tau: β=0.427, P<0.001), and there was a negative linear regression relationship between Q-alb and levels of Aβ 1-42 and Aβ 1-40 in cerebrospinal fluid (Aβ 1-42: β=-0.762, P<0.001; Aβ 1-40: β=-0.531, P<0.001). There was no linear regression relationship between Q-alb and level of p-Tau in group P ( P=0.121). There was no linear regression relationship between Q-alb and level of Aβ 1-40 in group NP ( P=0.467). The results of ROC curve analysis showed that the area under the curve for Q-alb in predicting POD (95% confidence interval) was 0.827 (0.738-0.896). Conclusion:Preoperative higher Q-alb is the risk factor for POD in patients undergoing neuraxial anesthesia, and is more accurate in predicting POD.
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Objective:To evaluate the changes in proteome in hippocampus and bioinformatics analysis in mice with perioperative neurocognitive disorders (PND).Methods:Clean-grade healthy male C57BL/6 mice, aged 15 months, weighing 30-35 g, were divided into 2 groups ( n=9 each) using a random number table method: control group (group C) and group PND.The model of PND was established by performing open tibial fracture with intramedullary fixation under isoflurane anesthesia in anesthetized mice.The Morris water maze test, open field test and fear conditioning test were performed at 1 day before operation and at 1, 3 and 7 days after operation.At 1, 3 and 7 days after operation, 3 mice with worst cognitive performance in each cognitive function assessments were sacrificed in group P, and three mice were randomly sacrificed in group C. The hippocampal tissues were then obtained, the expression of differentially expressed proteins was identified by high-performance liquid chromatography-mass spectrometry, and Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to analyze the differentially expressed proteins. Results:Compared with group C, the escape latency at different time points was significantly prolonged, and the percentage of time spend on target quadrant and the percentage of freezing time in fear conditioning test were decreased in group P ( P<0.05). There were 21 differentially expressed proteins, of which 12 proteins showed up-regulated expression and 9 proteins showed down-regulated expression.The GO functional analysis showed that the differentially expressed proteins were involved in the process such as the metabolism, signal transmission, regulation of biological processes, formed cell components such as synapses and organelles, and were related to molecular function such as binding and transportation.KEGG signaling pathway analysis showed that there were also differences in MAPK signaling pathway, ErbB signaling pathway, AMPK signaling pathway and the transport of SNARE protein in vesicle and etc. Conclusion:There are 21 differentially expressed proteins in the hippocampus of PND mice, and these proteins are involved in the pathophysiological process probably related to PND such as neuroinflammatory responses, abnormal synaptic structure, mitochondrial dysfunction and decreased autophagy.
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OBJECTIVE@#To analyze the molecular pathogenesis by analysis of phenotype and gene mutation in families with hereditary coagulation factor V (FⅤ) defect caused by complex heterozygous mutation.@*METHODS@#Plasma pro-thrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), FⅤ procoagulant activity (FⅤ∶C), FⅤ antigen (FⅤ∶Ag), and other related coagulation indexes were detected in the proband and his family members (3 generations 10 people). Using DNA direct sequencing to analyze all exons, flanks, 5' and 3' untranslated regions of F5 genes and the corresponding mutation site regions of family members, the mutation site was confirmed by reverse sequencing.The conservation of mutant amino acids was analyzed by ClustalX-2.1-win software. The PROVEAN and MutationTaster online bioinformatics software were used to predict the effect of mutation on protein function. Protein model and amino acid interaction at mutation sites was analyzed by Swiss-pdbviewer software.@*RESULTS@#The PT and APTT of the proband were significantly prolonged compared with healthy controls (34.2 vs 13.2 s and 119.3 vs 36.0 s), while FⅤ∶C and FⅤ∶Ag extremely reduced (3% and 6%). The PT and APTT of the second-born, the third son, daughter, and grandson of the proband were slightly prolonged, and the FⅤ∶C and FⅤ∶Ag decreased to varying degrees. The related coagulant parameters of other family members were within normal range. Genetic analysis revealed that the proband had a c.911G>A heterozygous missense mutation on the exon 6 lead to p.Gly276Glu, and a c.5343C>G heterozygous missense mutation on the exon 16 lead to p.Ser1781Arg of the proband. The second-born, the third son, and grandson of the proband carry p.Gly276Glu heterozygotes, and the daughter carries p.Ser1781Arg heterozygotes, while the other family members were wild-type. The results of conservative analysis indicated that p.Gly276 and p.Ser1781 were highly conserved in homologous species. The two bioinformatics software predicted the same results, PROVEAN (score -6.214 and -12.79) indicated that the compound heterozygous mutation was a harmful mutation; MutationTaster (score 0.976 and 0.999) suggested that these mutations might cause corresponding disease. p.Gly276Glu protein model analysis showed that, the Glu side chain was prolonged and the molecular weight became larger, which would increase the steric hindrance between it and the surrounding amino acids, affect the normal local folding of the FⅤ protein, and eventually lead to the decrease of protein activity and content. This paper can not provide analysis of the spatial structure of p.Ser1781Arg mutant protein because of the lack of X ray 3 D structure file of FⅤ exon 16.@*CONCLUSION@#The new compound heterozygous mutations (p.Gly276Glu and p.Ser1781Arg) identified in this study are the main reasons for the decrease in the FⅤ level of the family, among which p.Ser1781Arg is rarely reported at home and abroad.
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Humanos , Factor V/genética , Familia , Genotipo , Heterocigoto , Mutación , Linaje , FenotipoRESUMEN
<p><b>OBJECTIVE</b>To explore the effects and action mechanism of electroacupuncture (EA) pretreatment on rats with transient cerebral ischemia/reperfusion.</p><p><b>METHODS</b>A total of 144 healthy SD male rats were randomly divided into a sham operation group (group S), an ischemia/reperfusion group (group I/R) and an EA pretreatment group (group EA), 48 rats in each one. The model of cerebral ischemia/reperfusion was established by using 4-vessel occlusion method in the group I/R; after 5 min of cerebral ischemia, the reperfusion was performed. The group EA was treated with EA at "Dazhui" (GV 14) and "Baihui" (GV 20) 5 days before model establishment, 30 min per time, once a day. In group S, bilateral foramen alares were exposed without burning on the vertebral arteries, and bilateral common carotid arteries were unfolded and not occluded. The rats in the group I/R and group EA were sacrificed 6 h, 12 h, 24 h and 48 h after reperfusion and those in the group S were sacrificed at corresponding time to collect hippocampus example. The Western-blot method was used to measure the expression of glucose-regulated protein 78 (GRP 78), and HE staining method was used to count the number of surviving neurons, and TUNEL method was used to measure the number of apoptotic neurons.</p><p><b>RESULTS</b>Compare with the group S, the number of surviving neurons in hippocampus was reduced at each reperfusion time point and the number of apoptotic neurons was increased (all P<0.05) in the group I/R and the group EA; the expression of GRP 78 at each reperfusion time point in group I/R and group EA was increased (P<0.05). Compared with the group I/R, the number of surviving neurons in hippocampus was increased at each reperfusion time point and the number of apoptotic neurons was reduced in the group EA (P<0.05); the expression of GRP 78 at each reperfusion time point was further increased (P<0.05).</p><p><b>CONCLUSION</b>The electroacupuncture pretreatment has obvious cerebral protection on rats with ischemia/reperfusion, which is related with further increasing the expression of GRP 78 in ischemia area, leading to relieved endoplasmic reticulum stress.</p>
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Animales , Humanos , Masculino , Ratas , Isquemia Encefálica , Genética , Metabolismo , Cirugía General , Terapéutica , Electroacupuntura , Proteínas de Choque Térmico , Genética , Metabolismo , Hipocampo , Metabolismo , Ratas Sprague-Dawley , ReperfusiónRESUMEN
<p><b>OBJECTIVE</b>To screen potential mutation and explore the underlying mechanism for a consanguineous pedigree featuring hereditary coagulation factor Ⅴ (FⅤ) deficiency.</p><p><b>METHODS</b>Clinical diagnosis was validated by coagulant parameter assays of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), FⅤ procoagulant activity (FⅤ:C) and FⅤ antigen (FⅤ:Ag). Potential mutations of the F5 gene in the proband and his family members were analyzed by direct DNA sequencing of PCR products of all exons, exon-intron boundaries and 3', 5' untranslated regions. Suspected mutation was confirmed by reverse sequencing.</p><p><b>RESULTS</b>The PT and APTT in the proband were significantly prolonged, which measured 23.5 s (reference range 11.8-14.8 s) and 50.5 s (reference range 27.0-41.0 s), respectively. FⅤ activity and FⅤ antigen of the proband were significantly reduced to 8% and <1%, respectively. PT and APTT in the younger sister of the proband were also significantly prolonged (24.1 s and 62.4 s, respectively). Her FⅤ activity and FⅤ antigen were also significantly decreased (7% and <1%, respectively). PT and APTT of other family members were within the normal range. The homozygous missence mutation causing T→C transition at position 29170 in exon 5 of F5 gene has resulted in a Phe190Ser substitution in the proband. His younger sister was also homozygous for Phe190Ser. Heterozygosity for Phe190Ser was confirmed in his elder brother, elder sister, two daughters and niece, and their FⅤ activity were slightly decreased (57%, 73%, 72%, 66% and 75%, respectively). A normal wild type was observed in two younger brothers of the proband, and their FⅤ activity and FⅤ antigen were in the normal range.</p><p><b>CONCLUSION</b>Homozygous missence mutation of Phe190Ser has been found in above family featuring hereditary FⅤ deficiency. The homozygous missence mutation was inherited from the parents by consanguineous marriage. Phe190Ser probably underlies may underlie the pathogenesis of hereditary FⅤ deficiency in this pedigree.</p>
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Consanguinidad , Factor V , Genética , Deficiencia del Factor V , Sangre , Genética , Mutación Missense , Tiempo de Tromboplastina Parcial , Linaje , Tiempo de Protrombina , Análisis de Secuencia de ADNRESUMEN
<p><b>OBJECTIVE</b>To identify the genotype and pathogenesis in four Chinese pedigrees with Factor Ⅻ deficiency.</p><p><b>METHODS</b>Activated partial thromboplastin time (APTT), FⅫ procoagulant activity (FⅫ∶C), FⅫ antigen(FⅫ∶Ag)and other coagulant parameters were detected. The FⅫ deficiency Pedigree members,all exons,boundary introns including the splice junctions of the FⅫ gene were amplified with Polymerase chain reaction (PCR). Expression plasmids were constructed by mutagenesis based on the wild-type and transfected into COS7 cells. FⅫ∶C and FⅫ∶Ag of the expression levels were tested in the supernatant and cell lysate.</p><p><b>RESULTS</b>The four probands presented prolonged APTT with all the values of FⅫ∶C and FⅫ∶Ag were low to 2% and 1%, respectively. There were common 46C/T polymorphism in the promoter regions of FⅫ gene in four pedigrees. Proband A was heterozygous for two mutations, g.5741-5742delCA (His101Gln) and g.7142insertC (Lys346Gln). Proband B was a heterozygous deletion mutation g.6800-6808del9bp. The results of the transfection revealed that FⅫ∶Ag in cell lysates and conditioned media protein FⅫ6800-6808del9bp were 85.6% and 51.9%. The FⅫ∶C in the conditioned media was 56.4%. Proband C was a heterozygous mutation g.8699G>A(Gly542Ser). Proband D was a homozygous mutation 8699G>A, whose parents with consanguineous marriage.</p><p><b>CONCLUSION</b>Four mutations, g.5741-5742delCA, g.7142insertC, g.6800-6808del9bp and g.8699G>A with 46C/T polymorphism in the promoter regions of FⅫ gene, were identified in the four Factor Ⅻ deficiency pedigrees. The two mutations g.5741-5742delCA and g.6800-6808del9bp were first found in China. FⅫ 6800-6808del9bp expressed in vitro suggested that almost normal proteinum synthesis but defect proteinum secretion.</p>
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Adulto , Anciano , Femenino , Humanos , Lactante , Masculino , Análisis Mutacional de ADN , Factor XII , Genética , Deficiencia del Factor XII , Genética , Mutación , Linaje , Polimorfismo GenéticoRESUMEN
<p><b>OBJECTIVE</b>To observe the influence of auricular point sticking on incidence of nausea and vomiting and analgesia effect after gynecological laparoscopy, and provide evidence for clinical application of auricular point sticking.</p><p><b>METHODS</b>One hundred and twenty cases of selective gynecological laparoscopy under general anesthesia were randomly divided into an auricular point sticking group and a placebo group, 60 cases in each group. In the auricular point sticking group, the auricular point sticking with vaccaria seeds was applied at Shenmen (TF 4), Wei (CO 4) and Jiaogan (AH 6a) before the operation and 1, 5, 9, 23 h after the operation, which were pressed 5 min each point each time. The two ears were proceeded at the same time. In the placebo group, the same point selection, sticking paste was used as the auricular point sticking group, but no sticking or pressing with vaccaria seeds was adopted. The incidence of nausea and vomiting, the usage rate of tropisetron and morphine within 24 hours of the operation, as well as the score of visual analogue scale (VAS) and other adverse reactions at 2, 6, 10, 24 h after the operation were observed respectively.</p><p><b>RESULTS</b>Compared with the placebo group, the incidence of nausea and vomiting [31.7% (19/60), 16.7% (10/60) vs 58.3% (35/60), 35.0% (21/60)], the usage rate of tropisetron [21.7% (13/60) vs 48.3% (29/60)] and morphine [18.3% (11/60) vs 38.3% (23/60)], the VAS scores at all different time points in the auricular point sticking group were all decreased (all P < 0.05), and no adverse reaction was observed.</p><p><b>CONCLUSION</b>The auricular point sticking could significantly decrease the incidence of nausea and vomiting in patients of gynecological laparoscopy and has positive analgesic effect.</p>
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Adulto , Femenino , Humanos , Persona de Mediana Edad , Adulto Joven , Analgesia por Acupuntura , Puntos de Acupuntura , Acupuntura Auricular , Enfermedades de los Genitales Femeninos , Cirugía General , Ginecología , Laparoscopía , Náusea , Terapéutica , Vómitos , TerapéuticaRESUMEN
<p><b>OBJECTIVE</b>To analyze genetic mutation and explore its molecular pathogenesis for an hereditary protein C (PC) deficient consanguineous pedigree.</p><p><b>METHODS</b>The pedigree included three generations and contained eight members. PC activity (PC:A), PC antigen (PC:Ag) and other coagulant parameters were detected for all family members. Protein C gene (PROC) include all the exons and intron exon boundaries were amplified by PCR for the proband, then analyzed by direct sequencing. Mutation sites were detected for the other family members.</p><p><b>RESULTS</b>The PC:A and PC:Ag in the proband plasma were 20% (normal range 70% -140%) and 13.2% (normal range 70%-130%). A homozygous missense mutation g.6128T>G in exon 7 resulting in Phe139Val was identified in the proband. The PC:A and PC:Ag in her younger brother were 31% and 18.90%, Phe139Val homozygous was also found. The left family members were heterozygous for Phe139Val.</p><p><b>CONCLUSION</b>Phe139Val homozygous missense mutation in exon 7 of PROC caused serious hereditary protein C deficiency. We speculated that homozygous mutation might be resulted from this consanguineous marriage.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Consanguinidad , Homocigoto , Mutación , Linaje , Proteína C , Genética , Deficiencia de Proteína C , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To investigate potential mutations and clinical features of 9 unrelated patients with inherited coagulation factor VII (FVII) deficiency.</p><p><b>METHODS</b>Clinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, FVII activity and specific antigens. All exons, exon-intron boundaries, and 5' and 3' untranslated regions of F7 genes were amplified with PCR. Potential mutations were detected by direct sequencing of purified PCR products. Suspected mutations were confirmed by sequencing of the opposite strand.</p><p><b>RESULTS</b>All probands have featured prolonged prothrombin time, with FVII activity ranging between 2.0% to 6.0%. The titers of FVII antigen were significantly reduced in 7 probands. Eight mutations, including 6 missense mutations, 1 deletion and 1 insertion, were identified, among which 3 (Gln100Leu, Ser269Pro and g.11520_11521insT) were not described previously. Six mutations have located in the protease domain. All mutations were inherited, and consanguineous marriages were reported in 5 families. Mutations g.27_28delCT, Cys329Gly, Arg304Trp and His348Gln have been identified in unrelated families. There was a lack of correlation between the mutations and their clinical features. Two individuals with homozygous His348Gln mutations and 1 individual with homozygous Arg304Trp mutation were only mildly affected or asymptomatic. Two patients, who have respectively carried homozygous and heterozygous deletions of g.27_28delCT, were moderately affected and asymptomatic. In 4 patients carrying double heterozygous mutations, 1 (Ser269Pro and Cys329Gly) was asymptomatic, 2 (Arg304Trp and Cys329Gly, Arg277Cys and g.11520_11521insT, respectively) had a mild bleeding tendency, whilst 1 (Gln100Leu and His348Gln) has a moderate bleeding diathesis.</p><p><b>CONCLUSION</b>There seem to be hotspots of F7 gene mutations in ethnic Han Chinese populations. And there is a lack of correlation between particular types of mutations and clinical phenotypes.</p>
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Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Secuencia de Bases , Trastornos de la Coagulación Sanguínea Heredados , Genética , Factor VII , Genética , Deficiencia del Factor VII , Genética , Heterocigoto , Homocigoto , Datos de Secuencia Molecular , MutaciónRESUMEN
<p><b>OBJECTIVE</b>To analyze genetic mutations and explore its molecular pathogenesis for an hereditary protein C (PC) deficiency pedigree.</p><p><b>METHODS</b>The pedigree has included 15 individuals from 4 generations. Plasma levels of PC activity (PC:A), PC antigen (PC:Ag) and other coagulant parameters were determined for members of the family. The 9 exons and intron-exon boundaries of protein C gene (PROC) of the proband were amplified with PCR and analyzed with direct sequencing. Detected mutations were confirmed with reverse sequencing. Corresponding PCR fragments from the family members were also directly sequenced.</p><p><b>RESULTS</b>Plasma PC:A and PC:Ag for the proband was 26% and 18.60%, respectively, both being lower than normal references. Seven members from the pedigree also had lower PC:A, six had lower PC:Ag. A compound heterozygous missense mutation, including a T to G transition at position 6128 of exon 7, which results in Phe139Val, and a G to C transition at position 8478 in exon 9, which results in Asp255His, were identified in the proband. The paternal grandma, father and two aunts were heterozygous for g.6128 T to G, whilst the mother, the second uncle, sister and son were heterozygous for g.8478 G to C. There were lower PC:A in family members with g.8478 G to C.</p><p><b>CONCLUSION</b>The proband had inherited two independent mutations of the PROC gene including g.6128 T to G in exon 7 and g.8478 G to C in exon 9 from her father and mother, respectively. The resulting compound heterozygous mutation has caused a serious hereditary protein C deficiency.</p>
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Humanos , Mutación , Linaje , Proteína C , Genética , Deficiencia de Proteína C , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To study the efficacy and safety of patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in cesarean section.</p><p><b>METHODS</b>A randomized controlled clinical trail was conducted on 180 singleton primiparas (SAS > 30) undergoing selective cesarean section. They were randomly assigned to three groups, i. e., the patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) group (Group A, 60 cases), the patient-controlled sedation with transcutaneous electrical stimulation of auricular eye point group (Group B, 60 cases), and the control group (Group C, 60 cases). Patients in Group A received patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in the operating room. The strength was controlled by patients themselves. The stimulation lasted for 30 min before the epidural puncture till ending the surgery. Patients in Group B received stimulation of auricular eye point. Patients in Group C received pressurization with the same connected line as Group A, but without electric stimulation. The following indices were observed: (1) the bispectral index (BIS), heart rate (HR), mean arterial pressure (MAP), Ramsay sedation score when the women entered the operating room (T0), 30 min after stimulation (T1), at the time after removing the fetus (T2), and by the end of surgery (T3); (2) the concentrations of plasma angiotensin II (AngII) and cortisone (Cor) at the aforesaid time points; (3) the use rates of oxytocin, atropine, and ephedrine; the hemorrhage amount, and the neonatal Apgar score.</p><p><b>RESULTS</b>Compared with Group A, the BIS, the plasma concentrations of AngII and Cor increased at T1, T2, and T3 (P < 0.05), and the Ramsay sedation score decreased (P < 0.05). The HR and MAP increased at T1 (P < 0.05) in Group B and Group C. Compared with T0, the BIS, HR, MAP, and Ramsay sedation score, the plasma concentrations of AnglI and Cor were lowered in Group A at T1 (P < 0.05). There was no statistical difference in the use rates of oxytocin, atropine, and ephedrine; the hemorrhage amount, and the neonatal Apgar score (P > 0.05).</p><p><b>CONCLUSIONS</b>Patient-controlled sedation with transcutaneous electrical stimulation of auricular Shenmen (TF4) in cesarean section had obvious sedative effects. It had no adverse effects on puerperal or neonates.</p>
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Adulto , Femenino , Humanos , Embarazo , Puntos de Acupuntura , Analgesia Controlada por el Paciente , Métodos , Cesárea , Métodos , Dimensión del Dolor , Estimulación Eléctrica Transcutánea del Nervio , MétodosRESUMEN
<p><b>OBJECTIVE</b>To analyze genetic mutation and explore its molecular pathogenesis for an hereditary coagulation factor XII(F XII) deficiency in a pedigree featuring consanguineous marriage.</p><p><b>METHODS</b>Activated partial thromboplastin time (APTT), F XII procoagulant activity (F XII:C), F XII antigen (F XII:Ag) and other coagulant parameters were assayed. For the proband and his family members, exons 1-4, introns including the splice junctions of the F XII gene were amplified with polymerase chain reaction (PCR). The PCR product was purified and sequenced. The mutations were confirmed by sequencing the complimentary strand.</p><p><b>RESULTS</b>The proband has featured prolonged APTT at 157.5 s (reference range, 27.0-41.0 s). The APTT of his son has increased slightly at 48.3 s. The remaining members of the family were in normal range. F XII activity and F XII antigen of the proband were significantly decreased (<1%). The F XII activity of his wife, daughter, son and mother was also dropped to about 51%, 21%, 21% and 50%, respectively, and so was the F XII antigen (42%, 32%, 37% and 48%, respectively). Homozygous missense mutation of G→A transition at position 8699 in exon 14 resulting in Gly542Ser was identified in the proband. His mother, son and daughter were heterozygous for Gly542Ser. In the promoter regions of F XII gene, the genotype of the proband and the other members was 46T/T.</p><p><b>CONCLUSION</b>Homozygous missense mutation Gly542Ser was found in a pedigree of hereditary F XII deficiency. The homozygous missense mutation might have resulted from his parents by consanguineous marriage. Gly542Ser and 46T/T have contributed to the pathogenesis of the hereditary factor XII deficiency pedigree.</p>
Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Secuencia de Bases , Pruebas de Coagulación Sanguínea , Consanguinidad , Exones , Factor XII , Genética , Deficiencia del Factor XII , Sangre , Genética , Genotipo , Linaje , Polimorfismo de Nucleótido SimpleRESUMEN
<p><b>OBJECTIVE</b>To perform gene analysis and family survey of a patient with combined inherited FVII and FX deficiency, and to identify the gene mutation of this patient.</p><p><b>METHODS</b>The phenotype diagnosis was validated by coagulant parameter assay on prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FVII and FX activity (FVII:C, FX:C) and FVII and FX antigen (FVII:Ag, FX:Ag). FVII and FX gene mutations were analyzed in the proband and other family members by DNA direct sequencing of all exons, exon-intron boundaries and 5', 3' untranslated sequences. One hundred and six health examination participants were selected as control.</p><p><b>RESULTS</b>The values of PT and APTT of the proband showed significantly prolonged, which were 84.5s and 63.4s, respectively. The levels of FVII:C, FVII:Ag, FX:C and FX:Ag were 6%, 7%, 4% and 30%, respectively. The PT of his father, mother and sister was prolonged slightly while both APTT and FVII:Ag were in the normal range. Two homozygous mutations, g.11267C→T in exon 8 of FVII gene resulting in the substitution of Arg277Cys and g.28139G→T in exon 8 of FX gene leading to the substitution of Val384Phe, were identified in the proband. The proband's parents and sister were heterozygous for Arg277Cys and Val384Phe mutations.</p><p><b>CONCLUSION</b>Homozygous mutation Arg277Cys in FVII gene and Val384Phe in FX gene were the molecular mechanism causing combined inherited FVII and FX deficiency. The Val384Phe substitution was a novel mutation, which may affect the synthesis or secretion of FX protein.</p>
Asunto(s)
Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Secuencia de Bases , Análisis Mutacional de ADN , Factor VII , Genética , Deficiencia del Factor VII , Genética , Deficiencia del Factor X , Genética , Heterocigoto , Mutación , LinajeRESUMEN
<p><b>OBJECTIVE</b>To investigate the gene mutation and the molecular pathogenesis of an inherited coagulation factor VII (F VII) deficiency pedigree with consanguineous marriage.</p><p><b>METHODS</b>The diagnosis was validated by coagulant parameter assay on the prothrombin time (PT), activated partial thromboplastin time, fibrinogen and coagulation factor activity. F VII gene mutations were analyzed in the proband and other family members by direct DNA sequencing of the PCR products of all exons, exon-intron boundaries and 5'and 3' untranslated sequences. The mutations were confirmed by reverse sequencing.</p><p><b>RESULTS</b>The values of PT and F VII activity in the proband were significantly abnormal, they were 30.9 s and 3% respectively. The PT of her daughter, father and mother was slightly extended to 21.2 s, 16.3 s and 16.1 s respectively, and the F VII activity was reduced to 22%, 25% and 35% respectively. The coagulant parameters of her younger brother were within normal range. Homozygous T-->G transition at position 11482 in exon 8 was identified in the proband resulting in His348Gln, and heterozygosity for His348Gln was confirmed in her daughter and her parents, and the normal wild-type was observed in her younger brother.</p><p><b>CONCLUSION</b>Homozygous missense mutation of His348Gln was found in a pedigree of hereditary F VII deficiency. The mutation was inherited from her heterozygote parents.</p>
Asunto(s)
Adolescente , Adulto , Anciano , Femenino , Humanos , Recién Nacido , Masculino , Persona de Mediana Edad , Factor VII , Genética , Deficiencia del Factor VII , Genética , Homocigoto , Mutación Missense , LinajeRESUMEN
The objective of this study was to explore the changes of aggregation function of apheresis platelets and soluble P-selectin (sP-selectin) during storage. 20 samples of apheresis platelets were collected, and the aggregation function were examined by function test and the level of sP-selectin every day in storage of 5 days. The results showed that the aggregation function of platelets declined obviously during storage, there were significant differences between the first-day group and any of the other groups (p < 0.01). The max platelet aggregation rate was < or = 3% in the fourth-day group; sP-selectin level in plasma increased with prolong of storage time; there were significant differences between the first-day group and any of the other groups (p < 0.05). In conclusion, platelets were activated continuously during storage, while its aggregation function declines significantly. The ability of platelet aggregation to response to ADP loses almost completely since the fourth day during platelet storage. It should be paid more attention to the damage of apheresis collected platelets during storage.
Asunto(s)
Adulto , Humanos , Masculino , Plaquetas , Metabolismo , Fisiología , Selectina-P , Sangre , Agregación Plaquetaria , Recuento de Plaquetas , Plaquetoferesis , Métodos , Manejo de EspecímenesRESUMEN
The study was aimed to investigate the changes of blood coagulation factors during hemorrhagic shock in rats and the effects of various of resuscitation fluids on expression of blood coagulation factors in rats with hemorrhagic shock and to clarify its possible mechanism. 50 SD rats were randomly divided into 5 groups: control, sham operation, shock, resuscitation 1 (infusion with Ringer's lactate) and resuscitation 2 (infusion with 6% VOLUVEN), 10 rats per group. The rats in resuscitation 1 and resuscitation 2 groups were subjected to hemorrhagic shock, after hemorrhage shock for 1 hour resuscitation was performed with Ringer's lactate and 6% VOLUVEN. After resuscitation for 2 hours the changes of t-PA, PAI-1, TF were measured. At the same time, the rats in shock and the sham operation groups were blooded out so as to test. The results showed that the levels of plasma t-PA, t-PA/PAI, TF in the shock and resuscitation 1 groups were significantly higher than that in control and sham operation groups (P<0.01). The levels of plasma t-PA, t-PA/PAI in resuscitation 1 group were higher than that in shock group (P<0.01), the levels of plasma t-PA, t-PA/PAI and TF in the resuscitation 2 group were significantly lower than that in shock and resuscitation 1 groups (P<0.01). It is concluded that hemorrhagic shock may trigger the coagulation cascade reaction, results in hyperfunctioning of fiberinolysis and activation of platelets and coagulation system, and so the coagulation factor is greatly consumed. Unbalance of coagulation system plays an important role in the progress of shock. Efficacy of resuscitation with 6% VOLUVEN plus Ringer's lactate may be better than Ringer's lactate alone in regulating blood coagulation after hemorrhagic shock in rats.
Asunto(s)
Animales , Femenino , Masculino , Ratas , Factores de Coagulación Sanguínea , Metabolismo , Fluidoterapia , Inhibidor 1 de Activador Plasminogénico , Sangre , Distribución Aleatoria , Ratas Sprague-Dawley , Resucitación , Choque Hemorrágico , Sangre , Terapéutica , Tromboplastina , Metabolismo , Activador de Tejido Plasminógeno , SangreRESUMEN
<p><b>AIM</b>To study the effect of chronic hypoxic hypercapnia on expression of COX-2 mRNA in pulmonary arterioles.</p><p><b>METHODS</b>SD rats were randomly divided into two groups: control group and hypoxic hypercapnic group. COX-2 mRNA was observed in pulmonary arterioles by the technique of in situ hybridization.</p><p><b>RESULTS</b>mPAP, weight ratio of right ventricle (RV) to left ventricle plus septum (LV + S) and COX-2 mRNA in pulmonary arterioles were much higher in rats of hypoxic hypercapnic group than those of control group. Light microscopy showed that vessel smooth muscle cell hypertrophy and vessel cavity straightness were found in hypoxic hypercapnic group.</p><p><b>CONCLUSION</b>Changes of expressions of COX-2 mRNA may regulate hypoxic hypercapnic pulmonary hypertension.</p>