Objectives To detect the clinical significance of high sensitive C-reactive protein (hs-CRP) and immune function in children with Mycoplasma pneumoniae pneumonia (MPP). Methods 103 children with MPP, 47 cases of systemic inflammatory response syndrome (SIRS group), 56 cases of non-systemic inflammatory response syndrome (non-SIRS group) were recruited. 26 healthy children served as the control group. ELISA was used to detect the level of serum hs-CRP, immune in-dexes, IgG, IgA, and IgM, Cellular immune CD3+, CD4+, CD8+, CD4+/CD8+. Results The level of serum hs-CRP、IgG、IgM and CD8+in control group were significantly lower than those in non-SIRS group and SIRS group (P<0.05). The level of IgA、CD3+, CD4+, CD4+/CD8+in control group were significantly higher than those in non-SIRS group and SIRS group (P<0.05). The level of serum hs-CRP、IgG in SIRS group were significantly higher than those in non-SIRS group (P<0.05). The level of IgA、CD3+、CD4+、CD4+/CD8+in SIRS group were significantly lower than those in non-SIRS group. There was no significant difference in non-SIRS group and SIRS group of IgM and CD8+(P>0.05). The level change of serum hs-CRP were positively related with IgG (r=0.66,P=0.001) and were negatively related with IgA、CD4+、CD4+/CD8+(r=0.79, 0.67, 0.82, P all were<0.05) in chil-dren with MPP. Conclusion Children with MPP have Immunity function( including humoral immunity and cellular immunity) disorder which is related to the disease status. The level of hs-CRP could be an anpation index for the severity and immune func-tion of the children with MPP.

Objective To compare the effectiveness of microwave thermosphere ablation( M T A ) and traditional microwave ablation( M WA ) in ex v ivo bovine livers ,and to compare the degree of the heat sink effect in them .Methods T he non‐vessel model and vessel model were established using fresh bovine livers . In non‐vessel model ( n ＝ 48 ) ,the same power‐time settings were used in both M T A group and MWA group ,w hich were 80 W 12 min ,90 W 10 min and 100 W 10 min ,respectively . Long‐axis diameter ( Dl) , short‐axis diameter( Ds1 ,Ds2) ,roundness index ( R) and the time of temperature rising to 60℃ at place of 10 mm from the needle were measured .In vessel model( n ＝144) ,different vessel diameters( 3 mm ,5 mm ,10 mm) and flow rates ( 15 cm/s ,20 cm/s ,30 cm/s) were setted . The maximum radius( Rmax) ,total area of ablation zone( Sz ) ,and the area difference ( Sdiff ) were analyzed ,the temperatures adjacent to the vessel were monitored simultaneously . Results In non‐vessel model ,with the same power and time settings ,the Dl of M T A group was significantly smaller than MWA group ( P ＜ 0 .01 ) , however , there was no significant difference of Ds1 and Ds2 between the two groups( P ＞0 .05) . And M T A group created more spherical ablation zones ,since the R of M T A group were more close to 1 ( P ＜0 .01) . In M T A group ,the time of temperature rising to 60 ℃ at place of 10 mm from the needle was slower than MWA group ( P ＜0 .01) . In vessel model ,the Sdiff of M T A group were hardly affected by the vessel diameters and flow rates ( P ＞0 .05) ,and there was also no statistical significance among different flow rates( P ＞0 .05 ) in M WA group ,but the Sdiff of M WA group was significantly affected by the vessel diameters( P ＜0 .01) . And the Sdiff of M T A group was significantly smaller than M WA group when the vessel diameter was 10 mm ( P ＜0 .05) ,while there was no statistical significance between the two groups w hen the vessel diameter was 3 mm or 5 mm( P ＞0 .05) . Conclusions Compared to M WA ,M T A can produce sizable ,regular and more spherical ablation lesions with the same power and time ,meanwhile ,it is less affected by the heat sink effect .

OBJECTIVETo explore the effect and mechanism of sodium butyrate on expression of human clotting factor VIII in vitro.

METHODSMouse NIH/3T3 cell line was transfected with recombinant plasmid vector pRC/RSV-BDD-hFVIII, which enclosed B-domain deleted (760aa approximately 1 639aa) human factor VIII cDNA (BDD-hFVIII cDNA). Then cells were incubated in Dulbecco's modification of Eagle's medium (DMEM) containing sodium butyrate for 24 hours, hFVIII: C and hFVIII: Ag in the cell culture medium were measured by ELISA assay and one-stage method, respectively. In addition, the effect of sodium butyrate on transcription of cDNA encoding the whole hFVIII, heavy and light chain of hFVIII was also investigated by means of run-on assay.

RESULTSAfter stimulation of sodium butyrate, the levels of hFVIII: C and hFVIII: Ag increased 70% than those of control. Run-on assay showed that sodium butyrate enhanced the transcription of cDNA which encoded heavy chain of hFVIII.

CONCLUSIONSodium butyrate can improve the expression of hFVIII through enhancing the transcription of hFVIII heavy chain encoding cDNA. It demonstrated that sodium butyrate had potential utility in inducing the expression of hFVIII in vitro.

3T3 Cells ; Animals ; Butyrates ; pharmacology ; Factor VIII ; genetics ; Gene Expression Regulation ; drug effects ; Humans ; Mice

OBJECTIVETo investigate the plasma levels of coagulation factor VII (FVII) and polymorphisms of FVII gene in patients with coronary heart disease (CHD), and evaluate the effect of plasma FVII levels and FVII gene polymorphisms on CHD.

METHODSPlasma FVIIa, FVII: Ag and FVIIc were measured and polymorphisms of FVII gene were analyzed in 149 control cases and 60 CHD cases, including 33 acute myocardial infarction (AMI) cases by a combination of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and agarose gel electrophoresis.

RESULTSFVIIa, FVIIc in AMI group were significantly higher than that in control group, but FVII: Ag wasn't. There were no significant difference in plasma FVIIa, FVII: Ag and FVIIc between CHD and control group. The IVS7 genotypic frequency in AMI group was significantly different from that in control group. There was no significant difference in genotypic frequencies and allelic frequencies in other polymphism sites. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote.

CONCLUSIONSIncreased FVII levels, especially FVIIa and FVIIc in plasma, may contribute to coronary artery thrombosis. There was significant difference in IVS7 genotype frequency between control and AMI groups, but the rest weren't significantly different. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote. Polymorphism of -402 G/A may play an indirect role in AMI by regulating plasma FVII levels.

Coronary Disease ; blood ; genetics ; Factor VII ; analysis ; genetics ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

OBJECTIVETo accomplish a kind of therapeutic gene for hemophilia A, and observe the expression of human factor VIII (hF VIII) in vivo.

METHODSHuman clotting factor VIII cDNA with B-domain deleted (Delta760aa approximately 1639aa) was inserted into vector pRC/RSV to form pRC/RSV-hF VIII BD, which conjugated with in vivo liposome transfection reagent (DOTAP-Cholesterol) to accomplish a kind of therapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol. Mice were injected with pRC/RSV-hF VIII BD-DOTAP-Cholesterol i.m. and sacrificed 48 hours, 10 days, 20 days, 30 days, 40 days and 50 days later, respectively. Tissues such as heart, liver, spleen, lung, kidney and muscle were harvested, the distribution and transcription as well as expression of hF VIII BD cDNA were detected by means of PCR, RT-PCR and immunohistochemistry techniques. In addition, the antigen and antibody of hF VIII in plasma were measured.

RESULTSThere was high expression of hF VIII in plasma and tissues at the 48(th) hour after injection. On day 10, antigen level of hF VIII in plasma reached its peak, 17.55 ng/ml, and gradually reduced later. The antibody of hF VIII in plasma emerged on day 10 after injection, and increased and gradually reached 37.06 U/ml on day 50 after injection. PCR, RT-PCR and immunohistochemistry showed that hF VIII BD cDNA and its transcription as well as expression existed in all kinds of tissues, and lasted longer in spleen, lungs and kidneys than in heart, liver and muscle.

CONCLUSIONTherapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol, produced by combination of pRC/RSV-hF VIII BD and DOTAP-Cholesterol liposome can express human F VIII successfully in vivo, which lays an experimental foundation for curing hemophilia A by gene-drug in clinic.

Animals ; DNA, Complementary ; Disease Models, Animal ; Factor VIII ; biosynthesis ; genetics ; therapeutic use ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Hemophilia A ; therapy ; Humans ; Liposomes ; Mice ; Mice, Inbred BALB C ; Tissue Distribution ; Transfection

OBJECTIVETo identify the mutation of coagulation factor VII (F VII) gene in two pedigrees with hereditary F VII deficiency.

METHODSF VII gene mutations were analysed in two propositi and their family members by direct DNA sequencing. Allele specific PCR and PCR combined with restricted enzyme digestion were used to confirm the detected mutations.

RESULTSTwo gene mutations were detected in the propositus of pedigree A: G to C transition at position 6390 resulting in Trp40Cys and G to A at 11496 resulting in Arg353Gln, both are heterozygotes. The heterozygosity for polymorphism Arg353Gln was confirmed with the restriction enzyme Msp I digestion in his mother. In the propositus of pedigree B, there was a T to G transition at position 11482 resulting in His348Gln, heterozygosity of which was confirmed with Nsp I digestion in the propositus and his daughter. G to T transition at position 11514 resulting in Thr359Met was also found in the propositus of pedigree B, and the heterozygosity for Thr359Met was confirmed with allele specific PCR in the propositus and his son.

CONCLUSIONThree missense mutations were found in two pedigrees with hereditary F VII deficiency. A novel Trp40Cys mutation was reported for the first time.

Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation, Missense ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods

OBJECTIVETo explore gene defect of hereditary coagulation factor XIII deficiency.

METHODSPCR and gene sequencing or ARMS-PCR were used to detect the FXIIIA gene of peripheral white blood cell (PBC) from two Chinese hereditary coagulation factor XIII deficiency family members and 60 normal subjects respectively. The level of FXIIIA gene mRNA was tested by RT-PCR.

RESULTS(1) Nucleotide sequence analysis of the two probands' and their family members' DNA revealed that all of the three patients had homozygous missense mutation in FXIII A subunit gene. Proband 1 had a C to G transition at nucleotide (nt) 1 241 in exon 10 and proband 2 and his sister a C to T transition at nt 232 in exon 3 of FXIII A gene, which resulted in the substitution of Ser413 with Trp and Arg 77 with Cys, respectively. Family study showed that the two mutations were inherited from the parents who were correspondingly heterozygotes at nt 1 241 or nt 232. (2) The two mutations were not found in the normal subjects. (3) The FXIIIA gene mRNA level in the two probands was a little decreasing.

CONCLUSIONIt is the two novel mutations that results in FXIIIA deficiency. The two mutations of FXIIIA gene may affect its function or alter protein folding. The defective FXIII which is unstable and degraded rapidly in cytoplasm may be the main cause of FXIII deficiency.

Blood Coagulation Disorders, Inherited ; genetics ; Child ; Exons ; genetics ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods

Objective: To evaluate the incidence and mortality risk factors of pregnancy-related acute kidney injury (PR-AKI) in intensive care unit (ICU). Methods: A retrospective analysis was conducted. Critically ill pregnancies admitted to ICU of Shandong University Affiliated Provincial Hospital from January 1st, 2012 to December 31st, 2016 were enrolled. Based on the Kidney Disease: Improving Global Outcomes (KDIGO)-acute kidney injury (AKI) criteria, patients were divided into two groups: PR-AKI group and non-PR-AKI group. Clinical characteristics and laboratory data of two groups were compared. Risk factors of incidence and mortality of PR-AKI patients were analyzed, and the receiver operating characteristic (ROC) curve was drawn to evaluate the value of these risk factors in predicting mortality of PR-AKI patients in ICU. Results: ①A total of 219 pregnancies in ICU were included in the analysis, 85 cases (38.8%) were diagnosed with PR-AKI, with 29.4% in AKI stage 1, 27.1% in AKI stage 2 and 43.5% in AKI stage 3. ②Nineteen of 219 critically ill pregnancies died in ICU, the total ICU mortality was 8.7%. The mortality of PR-AKI group was higher than non-PR-AKI group (16.5% vs. 3.7%, P = 0.003). The mortality was worsened with increasing severity of AKI (4.0% for AKI stage 1, 4.3% for AKI stage 2, 32.4% for AKI stage 3). ③Acute fatty liver of pregnancy (AFLP) and lactate (Lac) were the independent risk factors for PR-AKI [AFLP: odds ratio (OR) = 6.081, 95% confidence interval (95%CI) was 1.587-23.308, P = 0.008; Lac: OR = 1.460, 95%CI was 1.078-1.977, P = 0.014]. ④ Age, Lac, acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) and sequential organ failure assessment (SOFA) were the independent risk factors associated with the mortality of PR-AKI patients in ICU (age: OR = 1.130, 95%CI was 1.022-1.249, P = 0.017; Lac: OR = 1.198, 95%CI was 1.009-2.421, P = 0.039; APACHEⅡ: OR = 1.211, 95%CI was 1.102-1.330, P < 0.001; SOFA: OR = 1.411, 95%CI was 1.193-1.669, P < 0.001). ⑤ ROC curve analysis showed that age, Lac, APACHEⅡscore and SOFA score all had good predictive values for in-hospital mortality among PR-AKI patients in ICU, the cut-off value was 29 years old, 3.8 mmol/L, 16 and 8, respectively, and the AUC was 0.751, 0.757, 0.892 and 0.919, respectively (all P < 0.01). Conclusions The incidence and mortality of PR-AKI of critically ill pregnancies in ICU are high. Increased age, Lac, APACHEⅡ score and SOFA score are independent risk factors associated with the mortality of PR-AKI patients in ICU, and have good predictive values for prognosis.