Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 86
Filtrar
Adicionar filtros








Intervalo de ano
1.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-776756

RESUMO

OBJECTIVE@#To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis.@*METHODS@#The FⅦ antigen (FⅦ:Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ:C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank.@*RESULTS@#The propositus had prolonged PT (36.3 s), with FⅦ:C and FⅦ:Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ:C (86%-120%). The FⅦ:Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein.@*CONCLUSION@#The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.


Assuntos
Fator VII , Genética , Deficiência do Fator VII , Genética , Feminino , Testes Genéticos , Heterozigoto , Humanos , Masculino , Mutação , Linhagem
2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-776737

RESUMO

OBJECTIVE@#To analyze the phenotype and F5 gene variant in a pedigree affected with hereditary coagulation factor V (FV) deficiency.@*METHODS@#All of the exons, flanking sequences, and 5' and 3' untranslated regions of the F5 gene were subjected to PCR and direct sequencing. Suspected variant sites were confirmed by clone sequencing. Influence of the variants was predicted by using software including ClustalX and Mutation Taster.@*RESULTS@#The prothrombin time (PT) and activated partial thromboplastin time (APTT) of the proband were prolonged to 20.3 s and 59.2 s, respectively, while FV activity (FV:C) and FV antigen (FV:Ag) were reduced by 13% and 17%, respectively. The FV:C and FV:Ag of his father, sister and daughter were decreased to 35%, 37%, 29% and 42%, 46%, 35%, respectively. The proband was found to carry a heterozygous c.2851delT variant in exon 13 of the F5 gene, which caused a frameshift and resulted in a truncated protein (p.Ser923LeufsX8). In addition, a heterozygous c.1538G to A (p.Arg485Lys) variant was found in exon 10. The father, sister and daughter of the proband all carried the p.Ser923LeufsX8 variant, while his mother and son carried the heterozygous p.Arg485Lys polymorphism. His younger brother and wife were of the wild type. Bioinformatic analysis showed that p.Ser923 was highly conserved across various species. Mutation Taster scored 1.00 for the p.Ser923LeufsX8 variant, and the result has predicted a corresponding disease.@*CONCLUSION@#A heterozygous deletional mutation c.2851delT in exon 13 of the F5 gene and a heterozygous c.1538G to A polymorphism harbored by the proband may be associated with the decreased FV level in this pedigree.


Assuntos
Fator V , Genética , Deficiência do Fator V , Genética , Feminino , Deleção de Genes , Testes Genéticos , Heterozigoto , Humanos , Masculino , Mutação , Linhagem , Fenótipo
3.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-800863

RESUMO

Objective@#To analyze the phenotype and F5 gene variant in a pedigree affected with hereditary coagulation factor Ⅴ (FⅤ) deficiency.@*Methods@#All of the exons, flanking sequences, and 5′ and 3′ untranslated regions of the F5 gene were subjected to PCR and direct sequencing. Suspected variant sites were confirmed by clone sequencing. Influence of the variants was predicted by using software including ClustalX and Mutation Taster.@*Results@#The prothrombin time (PT) and activated partial thromboplastin time (APTT) of the proband were prolonged to 20.3 s and 59.2 s, respectively, while FⅤ activity (FⅤ∶C) and FⅤ antigen (FⅤ∶Ag) were reduced by 13% and 17%, respectively. The FⅤ∶C and FⅤ∶Ag of his father, sister and daughter were decreased to 35%, 37%, 29% and 42%, 46%, 35%, respectively. The proband was found to carry a heterozygous c. 2851delT variant in exon 13 of the F5 gene, which caused a frameshift and resulted in a truncated protein (p.Ser923LeufsX8). In addition, a heterozygous c. 1538G>A (p.Arg485Lys) variant was found in exon 10. The father, sister and daughter of the proband all carried the p. Ser923LeufsX8 variant, while his mother and son carried the heterozygous p. Arg485Lys polymorphism. His younger brother and wife were of the wild type. Bioinformatic analysis showed that p. Ser923 was highly conserved across various species. Mutation Taster scored 1.00 for the p. Ser923LeufsX8 variant, and the result has predicted a corresponding disease.@*Conclusion@#A heterozygous deletional mutation c. 2851delT in exon 13 of the F5 gene and a heterozygous c. 1538G>A polymorphism harbored by the proband may be associated with the decreased FⅤ level in this pedigree.

4.
Chinese Journal of Anesthesiology ; (12): 1248-1252, 2019.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-797069

RESUMO

Objective@#To evaluate the effect of penehyclidine hydrochloride on the expression of airway mucin 5AC (MUC5AC) during ventilator-induced lung injury (VILI) and the relationship with Toll-like receptor 4/myeloid differentiation factor 88 (TLR4/MyD88) signaling pathway in rats.@*Methods@#Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-250 g, were divided into 3 groups (n=12 each) using a random number table method: sham operation group (group S), VILI group (group VILI), and penehyclidine hydrochloride group (group P). The rats were tracheotomized in group S. The rats were tracheotomized, connected to a small animal ventilator and mechanically ventilated for 4 h with the tidal volume of 20 ml/kg, respiratory rate 80 breaths/min, inspiratory/expiratory ratio 1∶1, and inspired oxygen fraction ratio 21% in VILI and P groups.At 30 min before mechanical ventilation, penehyclidine hydrochloride 2 mg/kg was injected via the tail vein in group P, and the equal volume of normal saline was given instead in S and VILI groups.At 4 h of mechanical ventilation, the arterial blood samples were taken for measurement of PaO2.The rats were then sacrificed, and broncho-alveolar lavage fluid (BALF) was collected for determination of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) concentrations by enzyme-linked immunosorbent assay.The lung specimens were collected for calculation of the wet/dry weight ratio (W/D ratio), for examination of pathological changes which were scored after haematoxylin and eosin staining (under a light microscope), and for determination of the expression of MUC5AC (by immunohistochemistry), expression of TLR4, MyD88, p38 mitogen-activated protein kinase (p38MAPK) and nuclear factor-kappa B (NF-κB) in lung tissues (by Western blot), and expression of MUC5AC mRNA in lung tissues (by real-time polymerase chain reaction).@*Results@#Compared with group S, PaO2 was significantly decreased, the W/D ratio and lung injury score were increased, the expression of MUC5AC protein and mRNA was up-regulated, the concentrations of IL-1β, IL-6 and TNF-α in BALF were increased, and the expression of TLR4, p38MAPK, MyD88 and NF-κB was up-regulated in VILI and P groups (P<0.01). Compared with group VILI, PaO2 was significantly increased, the W/D ratio and lung injury score were decreased, the expression of MUC5AC protein and mRNA was down-regulated, the concentrations of IL-1β, IL-6 and TNF-α in BALF were decreased, and the expression of TLR4, p38MAPK, MyD88 and NF-κB was down-regulated in group P (P<0.05).@*Conclusion@#Penehyclidine hydrochloride can decrease the expression of airway MUC5AC during VILI, and the mechanism may be related to inhibiting activation of TLR4/MyD88 signaling pathway in rats.

5.
Chinese Journal of Anesthesiology ; (12): 1168-1171, 2019.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-797050

RESUMO

Objective@#To compare the effect of total intravenous anesthesia with propofol and combined intravenous-inhalational anesthesia with sevoflurane on postoperative sleep quality in elderly female patients.@*Methods@#One hundred and twenty patients, aged 65-80 yr, with body mass index of 18.5-25.0 kg/m2, of American Society of Anesthesiologists physical status Ⅰ-Ⅲ, scheduled for elective gynecological laparoscopic operation, were divided into 2 groups (n=60 each) by a random number table method: total intravenous anesthesia with propofol group (group P) and combined intravenous-inhalational anesthesia with sevoflurane group (group S). Anesthesia was induced with intravenous etomidate 0.2-0.3mg/kg, sufentanil 0.2-0.4 μg/kg and cisatracurium 0.10-0.15 mg/kg.Anesthesia was maintained as follows: propofol was given by target-controlled infusion with the target plasma concentration set at 2-4 μg/ml in group P, 1.2-2.3% sevoflurane was inhaled in group S, and sufentanil 0.2-0.3 μg/kg and cisatracurium 0.04-0.06 mg/kg were intermittently injected in two groups.The patients were followed up at 7-8 a. m.on the day of hospitalization, the day of operation and 1, 3, 7 and 30 days after operation.The Pittsburgh Sleep Quality Index (PSQI) was recorded, PSQI > 7 points was considered as sleep disorders, and the occurrence of sleep disorders was recorded.The morning urine was collected at the time points mentioned above, and the concentrations of melatonin sulfate and free cortisol were determined by enzyme-linked immunosorbent assay.@*Results@#Compared with group S, the PSQI was significantly decreased on 3 and 7 days after operation, the incidence of sleep disorder was decreased on 7 days after operation, the concentrations of melatonin sulfate in urine were increased (P<0.05), and no significant change was found in free cortisol in urine in group P (P>0.05).@*Conclusion@#The effect of total intravenous anesthesia with propofol on postoperative sleep quality is less than that of combined intravenous-inhalational anesthesia with sevoflurane in elderly female patients.

6.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-796468

RESUMO

Objective@#To detect potential mutations of the coagulation factor Ⅶ (F7) gene in a pedigree affected with hereditary FⅦ deficiency and explore its molecular pathogenesis.@*Methods@#The FⅦ antigen (FⅦ∶Ag) was analyzed by an enzyme-linked immunosorbent assay (ELISA) method. Prothrombin time (PT), FⅦ activity (FⅦ∶C) and other coagulant parameters were quantified with an one-stage clotting assay. The F7 gene was amplified by PCR and sequenced. Mutational sites were confirmed by reverse sequencing. Impact of amino acid substitution was assessed using SIFT and PolyPhen-2 software. Structure of the mutant protein was analyzed using Swiss-pdb Viewer software based on the three-dimensional structure in the Protein Data Bank.@*Results@#The propositus had prolonged PT (36.3 s), with FⅦ∶C and FⅦ∶Ag significantly reduced to 2% and 44%, respectively. Her father, mother, younger sister and daughter had slightly prolonged PT and reduced FⅦ∶C (86%-120%). The FⅦ∶Ag of her father and younger sister were also reduced. DNA sequencing revealed that the propositus has carried compound heterozygous mutations (Lys341Glu and IVS6-1G>A) of the F7 gene. Her father and younger sister were heterozygous for the IVS6-1G>A mutation, while her mother and daughter were heterozygous for the Lys341Glu mutation. Bioinformatics analysis indicated that Lys341Glu mutation may affect the stability and function of the FⅦ protein.@*Conclusion@#The Lys341Glu and IVS6-1G>A mutations probably underlie the reduced activity of FⅦ in this pedigree.

7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-755645

RESUMO

Objective To evaluate the effects of selective brain hypothermia on the expression of miR-484 during cerebral ischemia-reperfusion ( I/R) in rats. Methods A total of 120 clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 4 groups ( n=30 each) using a random number table method: sham operation group ( group S) , cerebral I/R group ( group I/R) , hypothermia group ( group H) and normothermia group ( group N) . Blood vessels were only separa-ted and ligated without blockade in group S. In the other three groups, cerebral I/R was induced by inser-ting a nylon thread with rounded tip into the right internal carotid artery, and the middle cerebral artery was occluded for 2 h followed by reperfusion. In group H, 4℃ normal saline was infused for 15 min at a rate of 80 ml·kg-1 ·h-1 through the internal carotid artery immediately after removing the nylon thread. In group N, 37 ℃ normal saline was infused in the same way. Neurological deficit scores were evaluated at 6, 24 and 48 h after reperfusion. Animals were then sacrificed, the cerebral cortex of the ischemic penumbra was removed for determination of nerve cell apoptosis ( by TUNEL method) , expression of mitochondrial fission protein 1 ( Fis1) ( by Western blot) and expression of miR-484 ( quantitative real-time polymerase chain re-action) . The apoptotic rate was calculated. Results Compared with group S, the neurological deficit score and apoptotic rate of nerve cells were significantly increased, and the expression of Fis1 was up-regulated at each time point in the other three groups, the expression of miR-484 was significantly down-regulated in I/R and N groups, and the expression of miR-484 was significantly up-regulated in group H ( P<0. 05 or 0. 01) . Compared with group I/R and group N, the neurological deficit score and apoptotic rate of nerve cells were significantly decreased, and the expression of Fis1 was down-regulated, and the expression of miR-484 was up-regulated at each time point in group H ( P<0. 05 or 0. 01) . Conclusion The mechanism by which se-lective cerebral hypothermia attenuates cerebral I/R injury is associated with up-regulating miR-484 expres-sion and thus down-regulating Fis1 expression in rats.

8.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-755635

RESUMO

Objective To evaluate the role of α7 nicotinic acetylcholine receptor (α7nAChR) in postoperative cognitive dysfunction in aged rats with tibial fracture. Methods One hundred and fifty clean-grade healthy male Sprague-Dawley rats, aged 18-22 months, weighing 440-580 g, were divided into 5 groups ( n=30 each ) using a random number table method: control group ( group C ) , sham operation group ( group S) , tibial fracture group ( group T) , normal saline group ( group N) and α7nAChR agonist PUN282987 group (group P). Group C received no treatment. Ten percent chloral hydrate 0. 4 ml/100 g was injected intraperitoneally in group S. Group T underwent tibial fracture. PUN2829872. 4 mg/kg was in-traperitoneally injected at 5 min before tibial fracture in group P . The equal volume of normal saline was giv-en at 5 min before tibial fracture in group N. Morris water maze test was performed at day 7 after surgery. At days 1, 3 and 7 after surgery, the pathological changes of the hippocampal CA3 region were observed by haematoxylin and eosin staining, and the expression of α7nAChR, choline acetyltransferase ( ChAT ) , tumor necrosis factor-α( TNF-α) and interleukin-1β( IL-1β) in the hippocampal CA3 region was measured by Western blot. Results Compared with group C, the postoperative escape latency and swimming dis-tance were significantly prolonged, and the expression of α7nAChR, ChAT, TNF-α and IL-1β was up-regulated at each time point after operation in T, N and P groups ( P<0. 05) , and no significant change was found in the parameters mentioned above in group S ( P>0. 05) . Compared with group T, the postoper-ative escape latency and swimming distance were significantly shortened, and the expression of α7nAChR and ChAT was up-regulated and the expression of TNF-α and IL-1β was down-regulated at each time point after operation in group P ( P<0. 05) , no significant change was found in the parameters mentioned above in group N ( P>0. 05) , and the pathological changes of the hippocampal CA3 region were significantly at-tenuated in group P. Conclusion α7nAChR antagonism is involved in the development of postoperative cognitive dysfunction in aged rats with tibial fracture.

9.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-755546

RESUMO

Objective To evaluate the changes in the expression of hippocampal long non-coding RNAs ( lncRNAs) and bioinformatics analysis in mice with perioperative neurocognitive disorders ( PND) . Methods Thirty clean-grade male C57BL∕6 mice, aged 12-14 weeks, weighing 25-30 g, were divided into 2 groups (n=15 each) according to the random number table method: control group (group C) and PND group. The model of PND was established by performing open tibial fracture with intramedullary fixa-tion under isoflurane anesthesia in anesthetized mice. The Morris water maze test, open field test and fear conditioning test were performed at 1, 3 and 7 days postoperatively. The animals were sacrificed after the end of behavioral testing on 3 days after operation, the hippocampus was obtained, the high-throughput gene sequencing was performed to identify the differentially expressed lncRNAs, and Gene Ontology func-tional analysis and Kyoto Encyclopedia of Genes and Genomes signaling pathway analysis were used to ana-lyze the results. Results Compared with group C, the escape latency was significantly prolonged, and the percentage of time spend in the target quadrant and percentage of freezing time in the fear conditioning test were decreased at different time points after operation in group PND ( P<0. 05) . A total of 121 differential-ly expressed lncRNAs were identified, of which 69 were up-regulated and 52 were down-regulated. The Gene Ontology functional analysis showed that there were differences in various biological processes, such as synaptic transmission, cholinergic neurotransmitters, or adiponectin secretion and regulation. The KEGG signaling pathway analysis showed that there were also differences in cholinergic synapses, MAPK signaling pathway, glucagon signaling pathway, TNF signaling pathway, NOD-like receptors, Toll-like re-ceptors, chemokine signaling pathway and etc. Conclusion There are 121 differentially expressed lncR-NAs in the hippocampus of PND mice, and lncRNAs- and the target gene-related inflammatory responses, synaptic transmission, energy metabolism and etc. may be related to the pathogenesis of PND.

10.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-755526

RESUMO

Objective To evaluate the effect of different approaches to thoracolumbar interfascial plane (TLIP) block on the analgesic efficacy after lumbar spinal fusion.Methods Seventy-five patients of both sexes,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 40-70 yr,with body mass index ≤40 kg/m2,undergoing posterior median approach to lumbar spinal fusion,were divided into 3 groups (n =25 each) using a random number table method:patient-controlled intravenous analgesia (PCIA) group (group P),medial approach to TLIP combined with PCIA group (group MP) and lateral approach to TLIP combined with PCIA group (group LP).TLIP block was performed by advancing the block needle in a lateral to medial direction and injecting 0.5% ropivacaine 20 ml locally between the bilateral multifidus and longissimus muscles in group MP.TLIP block was performed by advancing the block needle in a medial to lateral direction and injecting 0.5% ropivacaine 20 ml locally between the bilateral longissimus and iliocostalis muscles in group LP.The identification rate and distinction score between multifidus and iliocostalis muscles obtained from ultrasonic images were recorded before block in MP and LP groups.All the patients received PCIA until 48 h after surgery.PCIA solution contained sufentanil 100 μg and tropisetron 10 mg diluted to 100 ml with 0.9% normal saline.The PCA pump was set up with a 2 ml bolus dose,a 15-min lockout interval and background infusion at a rate of 1 ml/h.When visual analogue scale score at rest >4 after surgery,parecoxib 40 mg was intravenously injected for rescue analgesia.The development of adverse reactions such as nausea and vomiting was recorded within 48 h after surgery.Results The identification rate and distinction score of the iliocostalis was significantly higher than that of the multifidus (P<0.05).Compared with group P,the intraoperative consumption of propofol and remifentanil was significantly decreased,the pressing times of PCA,amount of drugs consumed in PCA and consumption of parecoxib were decreased within 48 h after surgery (P<0.05),and no significant change was found in the incidence of nausea or vomiting in MP and LP groups (P>0.05).There was no significant difference in the parameters mentioned above between group MP and group LP (P>0.05).Conclusion Ultrasound-guided medial and lateral approaches to TLIP block is helpful in achieving low-dose opioid anesthesia mode and more helpful in enhancing the efficacy of PCIA when used for lumbar spinal fusion.

11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-745658

RESUMO

Objective To evaluate the effect of electro-acupuncture preconditioning on the expression of protein kinase R-like endoplasmic reticulum kinase (PERK) in the hippocampus of mice following cerebral ischemia-reperfusion (I/R).Methods A total of 120 healthy male C56BL/6 mice,aged 7 weeks,weighing 20-30 g,were divided into 3 groups (n=40 each) using a random number table method:sham operation group (group S),cerebral I/R group (group I/R) and electro-acupuncture preconditioning group (group EA).Cerebral I/R was induced by clamping bilateral common carotid arteries for 15min followed by restoring cerebral blood flow in anesthetized mice.Dazhui and Baihui acupoints were stimulated with disperse-dense waves of 1 mA for 30 min once a day,lasting for 5 consecutive days,and cerebral I/R model was established at 24 h after the last stimulation in group EA.Animals were sacrificed at 24and 48 h of reperfusion,brains were removed,and hippocampi were isolated for determination of apoptotic neurons in hippocampal CAl region (by TUNEL) and expression of PERK (by Western blot).Results Compared with group S,the neurological deficit score and the number of apoptotic neurons in hippocampal CAl region were significantly increased at each time point of reperfusion in I/R and EA groups (P<0.05).Compared with group I/R,the neurological deficit score and the number of apoptotic neurons in hippocampal CAl region were significantly decreased at each time point of reperfusion in group EA (P<0.05).Conclusion The mechanism by which electro-acupuncture preconditioning mitigates endoplasmic reticulum stress during cerebral I/R may be related to inhibiting PERK activity in mice.

12.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-694917

RESUMO

Objective To evaluate the effect of extremity ischemia preconditioning on lung protection in patients undergoing pulmonary lobectomy,and to further explore its possible mecha-nisms.Methods Forty patients (male 26 and female 14,BMI 20-28 kg/m2,ASA Ⅰ or Ⅱ,aged 45-64 years old)scheduled for elective pulmonary lobectomy via a thoracoscope,were randomly divided into two groups:extremity ischemic preconditioning group (group P)had the left leg roots tied with a tourniquet,lower extremity blood flow aerated blocked for 5 min,then deflated to restore blood flow for 5 min,the cycle was repeated three times;control group (group C)had the left lower extremity tied but not charged for 30 min.All the patients underwent intravenous-inhaled composite anesthesia. Arterial and venous blood samples were taken after admission to the operating room,at 6 h,12 h and 24 h after operation.The alveolar-arterial oxygen pressure difference (PA-aO2),respiratory index(RI) and oxygenation index (OI)were calculated.The level of TLR4 was measured.The pulmonary com-plications within 48 h after operation were recorded.The adverse reactions of the left lower extremity of the group P were recorded.Results Compared with the admission to the operating room,the ex-pression of TLR4 was significantly increased in the two groups at 6 h,12 h and 24 h after operation, but the expression of the group P was significantly lower than that of group C (P<0.05).Compared with the admission to the operating room,PA-aO2and RI were significantly increased,and OI was sig-nificantly decreased in the two groups at 6 h,12 h and 24 h after operation (P<0.05).Compared with group C,PA-aO2and RI were significantly decreased,and OI was significantly increased in group P (P<0.05).There was no statistically significant difference in the incidence of pneumonia and ate-lectasis in the two groups after operation,and no respiratory failure was observed in both groups.The left lower extremity of the group P had no adverse reactions,such as rupture of skin,thromboembo-lism,and nerve injury.Conclusion Extremity ischemic preconditioning has protective effect on pul-monary function in the patients undergoing pulmonary lobectomy,which may be related to down-reg-ulation of TLR4 expression in monocytes of blood and inhibition of the systemic inflammatory re-sponse.

13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-712148

RESUMO

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)

14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-712129

RESUMO

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.

15.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-709842

RESUMO

Objective To evaluate the effect of selective brain hypothermia on small ubiquitin-like modifier (SUMO)ylation of cerebral cortex during cerebral ischemia-reperfusion (I/R) in rats.Methods A total of 120 healthy clean-grade male Sprague-Dawley rats,aged 8 weeks,weighing 200-250 g,were divided into 4 groups (n=30 each) by a random number table method:sham operation group (group S),group I/R,selective brain hypothermia group (group H) and 37 ℃ normal saline group (group N).Rats were anesthetized with 10% chloral hydrate 3 ml/kg.Cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into the right internal carotid artery and advanced cranially until resistance was met.The right middle cerebral artery was occluded for 2 h followed by reperfusion in group I/R.10-13 ℃ normal saline was infused at the rate of 100 ml · kg-1 · h-1 for 20 min starting from the time point immediately after removing the nylon thread in group H.37 ℃ normal saline was infused instead at the same rate for 20 min in N group.The neurological deficit were assessed and scored at 6,24 and 48 h after reperfusion.The cerebral cortex on ischemic side was then removed for determination of cell apoptosis (by TUNEL) and expression of SUMO sepcific proteases 3 (SENP3) and SUMO2/3-conjugated protein (by Western blot).The apoptotic rate was calculated.Results Compared with group S,the neurological deficit score and apoptosis rate were significantly increased,and the expression of SUMO2/3-conjugated protein was up-regulated at each time point after reperfusion in the other three groups (P<0.05),and the expression of SENP3 was significantly up-regulated in I/R and N groups and down-regulated in group H (P<0.01).Compared with group I/R,the neurological deficit score and apoptosis rate were significantly decreased,the expression of SUMO2/3-conjugated protein was up-regulated,and the expression of SENP3 was down-regulated in group H (P<0.05).Conclusion The mechanism by which selective brain hypothermia reduces cerebral I/R injury is related to enhancing SUMOylation of cerebral cortex in rats.

16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-709729

RESUMO

Objective To evaluate the effect of electroacupuncture (EA) preconditioning on inositol-requiring kinase 1 (IRE1)-X-box binding protein 1 (XBP1) signaling pathway in endoplasmic reticulum in the cortex in a rat model of cerebral ischemia-reperfusion (I/R).Methods One hundred and eight pathogen-free healthy male Sprague-Dawley rats,aged 8-12 weeks,weighing 200-250 g,were assigned into 3 groups (n =36 each) using a random number table:sham operation group (group S),group I/R and EA preconditioning group (group EA).Focal cerebral I/R was induced by occlusion of right middle cerebral arteries for 2 h followed by reperfusion in rats anesthetized with chloral hydrate.In group EA,Baihui acupoints were stimulated with an electric stimulator for 30 min once a day for 5 consecutive days starting from 5 days before ischemia,and the model was established at 24 h after the last preconditioning.Rats were sacrificed after neurological deficit was scored at 6,12 and 24 h of reperfusion,brains were removed,and the ischemic area in cerebral cortex was isolated for examination of the cell ultrastructure (with an electronic microscope) and for determination of the expression of IRE1 and XBP1 (by Western blot).Results Compared with group S,the neurological deficit scores were significantly increased,and the expression of IRE1 and XBP1 in the ischemic area was up-regulated at each time point in I/R and EA groups (P<0.01).Compared with group I/R,the neurological deficit scores were significantly decreased,and the expression of IRE1 and XBP1 was up-regulated at each time point in group EA (P<0.05).The cell damage in the ischemic area in cerebral cortex was significantly attenuated in group EA when compared with group I/R.Conclusion The mechanism by which EA preconditioning attenuates cerebral I/R injury is related to activating IRE1-XBP 1 signaling pathway and relieving endoplasmic reticulum stress in rats.

17.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-709691

RESUMO

Objective To evaluate the effect of perioperative continuous femoral nerve block on postoperative cognitive dysfunction(POCD)in elderly patients with femoral neck fractures. Methods Sixty American Society of Anesthesiologists physical statusⅡorⅢpatients of both sexes, aged 65-90 yr, scheduled for femoral head replacement under combined spinal?epidural anesthesia, were divided into 2 groups(n=30 each)using a random number table: patient?controlled femoral nerve block for analgesia (PCNA)group and patient?controlled intravenous analgesia(PCIA)group. When the patients were diag?nosed with femoral neck fractures after admission to hospital in two groups, patients received continuous PCNA under nerve stimulator guidance until 48 h after operation in group PCNA, and fentanyl was continu?ously injected for analgesia until 48 h after operation in group PCIA. Neuraxial anesthesia was performed at L3,4interspace, 2 ml cerebrospinal fluid was extracted after successful subarachnoid puncture to detect the concentrations of Aβ?42 and tau, and the ratio of Aβ?42 to tau was calculated. The patient′s cognitive func?tion was assessed using Mini?Mental State Examination before analgesia and at 7 days after operation, and the occurrence of POCD was recorded. Results The ratio of Aβ?42 to tau in cerebrospinal fluid was signif?icantly lower, and the incidence of POCD was higher at 7 days after operation in group PCIA than in group PCNA(P<0.05). Conclusion Perioperative continuous femoral nerve block can decrease the develop?ment of POCD in elderly patients with femoral neck fractures.

18.
Chinese Journal of Anesthesiology ; (12): 1146-1149, 2018.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-734641

RESUMO

Objective To investigate the effect of acupuncture on cholinergic anti-inflammatory pathway in the hippocampus of aged rats during global cerebral ischemia-reperfusion ( I∕R ) . Methods Ninety-six clean-grade healthy male Sprague-Dawley rats, aged 17-22 months, weighing 440-580 g, were divided into 3 groups ( n=32 each) using a random number table method: sham operation group ( group S), global cerebral I∕R group (group I∕R) and acupuncture group (group AP). Global cerebral I∕R was induced by 4-vessel occlusion method described by Pulsinelli in group I∕R and group AP. Baihui and Feng-chi were stimulated for 14 consecutive days before ischemia in group AP. Four rats were sacrificed at 1, 3, 5 and 7 days of reperfusion, and brains were removed for determination of neuronal apoptosis by TUNEL. Four rats were sacrificed at 1, 3, 5 and 7 days of reperfusion, and brains were removed for determination of the expression of α7 nicotinic acetylcholine receptor (α7nAChR), choline acetyltransferase (ChAT), tumor necrosis factor-α ( TNF-α) and interleukin-1β ( IL-1β) in the hippocampal CA1 region by Western blot. The apoptosis rate was calculated. Results Compared with group S, the apoptosis rate of hippocam-pal neurons was significantly increased, and the expression of α7nAChR, ChAT, TNF-α and IL-1β was up-regulated at each time point of reperfusion in I∕R and AP groups ( P<0. 05) . Compared with group I∕R, the apoptosis rate of hippocampal neurons was significantly decreased, the expression of α7nAChR and ChAT was up-regulated, and the expression of TNF-α and IL-1β was down-regulated at each time point ofreperfusion in group AP (P<0. 05). Conclusion The mechanism by which acupuncture mitigates global cerebral I∕R injury may be related to activating cholinergic anti-inflammatory pathway in the hippocampus of aged rats.

19.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-734595

RESUMO

Objective To evaluate the efficacy of oxycodone combined with thoracic paravertebral block ( TPVB) for postoperative analgesia in patients undergoing minimally invasive direct coronary artery bypass grafting ( MIDCABG) . Methods Thirty-two American Society of Anesthesiologists physical statusⅡorⅢpatients of both sexes, aged 60-75 yr, weighing 50-85 kg, scheduled for elective MIDCABG un-der general anesthesia, were divided into 2 groups ( n=16 each) using a random number table method:morphine plus TPVB group ( group MT) and oxycodone plus TPVB group ( group OT) . Paravertebral cathe-ter was placed at T4,5 before induction of anesthesia to perform left thoracic paravertebral puncture, patients were tracheally intubated, and 0. 375% ropivacaine 15 ml was injected followed by continuous infusion of 0. 375% ropivacaine 5 ml∕h until 0. 5 h before the end of surgery. Both groups received patient-controlled analgesia ( PCA) after surgery. The PCA solution contained 1 mg∕ml morphine 60 ml in group MT or 1 mg∕ml oxycodone 60 ml in group OT, and the PCA pump was set up to deliver a 1 mg bolus dose with a 10-min lockout interval and background infusion at 1 ml∕h after a loading dose of 2 mg, with the maximum dose of 20 mg every 4 h. Pethidine 50 mg was intravenously injected as a rescue analgesic to maintain visual ana-log scale≤4. The intraoperative consumption of fentanyl, consumption of analgesics for PCA within 48 h after surgery, ratio of total to effective pressing times of PCA, consumption of analgesics for rescue analge-sia, requirement for rescue analgesia, score of satisfactory analgesia, extubation time, duration of inten-sive care unit stay and length of hospital stay were recorded. The development of nausea and vomiting, pru-ritus, respiratory depression, atelectasis and somnolence was recorded within 72 h after surgery. Results Compared with group MT, the intraoperative consumption of fentanyl, consumption of analgesics for PCA, consumption of analgesics for rescue analgesia, requirement for rescue analgesia and ratio of total to effec-tive pressing times of PCA were significantly decreased, the score of satisfactory analgesia was increased, the extubation time and duration of intensive care unit stay were shortened, and the incidence of nausea and vomiting, pruritus, respiratory depression and somnolence was decreased in group OT (P<0. 05). Con-clusion Oxycodone combined with TPVB provides safe and effective efficacy for postoperative analgesia in patients undergoing MIDCABG.

20.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wprim-687983

RESUMO

<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>


Assuntos
Adulto , Afibrinogenemia , Genética , Feminino , Fibrinogênio , Química , Genética , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Linhagem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA