Background Bone marrow-derived mesenchymal stem cells (MSCs) have been reported to have the ability of migrating after transplantation into rat model of traumatic brain injury. In this study, MSCs'characteristic of tropism for intracranial glioma was explored following being labeled and transplanted into brain and blood of gliomabearing rats.Methods Cellulae medullares were cultured to get pure MSCs. The cell surface antigen and cell cycle of MSCs were detected to confirm their identity by flow cytometry. MSCs were marked with BrdU and then were injected into contralateral brain or collateral internal carotid artery of rats bearing glioma. 2 weeks later, the brains were resected and pathological sections were made. Immunohistochemistry and immunofluorescence technology were performed to detect MSCs.Results MSCs displayed extensive tropism for intracranial glioma. MSCs which were injected in the contralateral brain of the glioma scattered densely in the juncture between brain tissue and tumor and there were a few MSCs in the glioma; MSCs which were injected into the collateral internal carotid artery scattered widespread in the glioma and there were a few MSCs in the juncture area.Conclusions MSCs have the ability to migrating toward glioma and penetrating blood brain barrier. MSCs may be ideal gene therapy vehicles against glioma.

OBJECTIVE To study the distribution of drug-resistant gene of meticillin-resistant coagulase negative Staphylococcus(MRCNS) in children septicemia.METHODS The total MRCNS isolates were 40,and whether in there harbored genes mec,erm and qac was studied.RESULTS Among 40 CNS strains,in there harbored genes mec,erm,and qac were 38(95.0%),30(75.0%) and 18(45.0%),respectively.CONCLUSIONS MRCNS in children septicemia where harbor drug resistance genes is very serious,so we should pay great heed to its effective control.

OBJECTIVE To investigate dynamically antibiotic resistance changing of Escherichia coli in Hubei province and provide the reference for clinical rational application of antibiotics.METHODS The WHONET4-5 software was used to analyze the antibiotic resistant rates of 2901 strains of E.coli isolated from all kinds of clinical specimen of children from 2003 to 2006 in the third grade hospitals of Hubei province.RESULTS Drug-resistance rate of E.coli in children group was lower than that in adult group.Except for 100.0% susceptibility to carbopenems such as imipenem and Meropenem,in the four years,drug-resistance of the other antibiotics showed ascending tendency.Ampicillin showed the highest drug-resistance rate(84%-90%).The lower(3%-25%) were amikacin,cefoxitin,ceftazidime,cefepime.CONCLUSIONS Antibiotic resistance of E.coli in children in Hubei province showed the ascending tendency.We should take effective measurement to control the spread and prevalence of the drug-resistant bacteria.

OBJECTIVE To approach biosafety risks in laboratory department of children′s hospital to formulate the managemen strategy.METHODS The existing biosafety risk in laboratory department of children′s hospital was investigated and analyzed.RESULTS A series of problems such as personnel′s biosafety consciousness,work environment,protection equipment,a medical orderly′s training,etc were existing.CONCLUSIONS Biosafety regulations should be executed strictly in children′s hospital,and biosafety management should be highly paid attention to and consummated.

Objective:To investigate the influence of bundled ligation of the pancreatic stump on postoperative pancreatic fistula (POPF) of distal pancreatectomy (DP).Methods:The retrospective case-control study was conducted. The clinical data of 60 patients with diseases in pancreatic body and tail who underwent DP in the Affiliated Hospital of Inner Mongolia Medical University from January 2011 to August 2018 were collected. There were 24 males and 36 females, aged from 19 to 68 years, with a median age of 45 years. Of the 60 patients, 36 cases undergoing dissection of pancreas with Endo-GIA stapler were allocated into non-bundled group, and 24 cases undergoing bundled ligation of the pancreatic stump with No.10 or No.7 suture at the site over 1 cm of the resection site before dissection of pancreas were allocated into bundled group. Observation indicators: (1) postoperative situations; (2) analysis of risk factors for POPF of DP. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test or ANOVA test. Measurement data with skewed distribution were represented as M (range).Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the chi-square test and multivariate analysis was conducted using the Logistic regression model. Results:(1) Postoperative situations: the amylase concentration, cases with biochemical fistula, cases with grade B pancreatic fistula, cases with complications, time to extubation, duration of postoperative hospital stay, total hospital expenses were (2 629±592)U/L, 14, 5, 7, (11.9±0.7)days, (13.6±0.7)days, (49 430±1 626)yuan in non-bundled group and (683±312)U/L, 3, 1, 2, (9.7±0.6)days, (11.3±0.5)days, (44 767±1 163)yuan in bundled group, respectively. There were significant differences in the amylase concentration, cases with biochemical fistula, time to extubation, duration of hospital stay, total hospital expenses between the two groups ( t=2.528, χ2=1.512, t=2.341, 2.311, 2.111, P<0.05), and there was no significant difference in the cases with grade B pancreatic fistula or cases with complications between the two groups ( χ2=1.512, 1.394, P>0.05). (2) Analysis of risk factors for POPF of DP. Results of univariate analysis showed that tumor diameter and bundled ligation of the pancreatic stump were related factors of patients undergoing pancreatic fistula after DP ( χ2=4.462, 5.061, P<0.05). Results of multivariate analysis showed that bundled ligation of the pancreatic stump was an independent influencing factor of patients undergoing pancreatic fistula after DP ( odds ratio=0.187, 95% confidence interval as 0.037-0.954, P<0.05). Conclusions:Bundled ligation of the pancreatic stump was an independent influencing factor of patients undergoing pancreatic fistula after DP. Bundled ligation of the pancreatic stump can effectively reduce the incidence of POPF, especially biochemical fistula, the time to extubation, duration of postoperative hospital stay and total hospital expenses, and promote patient recovery after DP.

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