Objective To investigate the changes in intrapulmonary shunting during controlled hypotension induced by sodium nitroprusside(SNP)in patients undergoing naso-endoscopic operation.Methods Forty ASAⅠorⅡpatients of both sexes(23 male,17 female)aged 16-50 yrs weighing 50-75 kg undergoing naso-endoscopic operation under general anesthesia with muscle relaxation and mechanical ventilation were studied.Radial artery was cannulated for direct BP monitoring and blood sampling.Right internal jugular vein was cannulated and the catheter was advanced into right ventricle.Blood sample taken from right ventricle was used as mixed venous blood instead of blood from pulmonary artery.ECG,MAP,HR and P_(ET) CO_2 were continuously monitored during operation Cardiac output was monitored with noninvasive cardiac function monitor(NC-COM.)based on impedance principle.SNP infusion was started at the beginning of operation at 1-3?g?kg~(-1)?min~(-1) and was then adjusted.MAP was reduced by 30%-40% and maintained at this level until the end of operation.Blood samples were taken from artery and right ventricle simultaneously before SNP infusion(T_1,baseline)at 30 and 60 min of hypotension(T_2,T_3)and at 20 min after BP returned to the baseline level(T_4)for blood gas analysis.Qs/Qt was calculated.Results Qs/Qt was significantly increased during controlled hypotension at T_2 and T_3 as compared to the baseline value(P＜0.01)and returned to the baseline level at T_4.HR was increased and cardiac output and stroke volume was significantly reduced during hypotension as compared to the baseline value.Conclusion The intrapulmonary shunting is increased and the hemodynamics is depressed during SNP-induced controlled hypotension and they return rapidly to baseline level after SNP is discontinued.No hypoxemia develops during SNP- induced hypotension.

Objective To identify gene mutations in two families with epidermolytic hyperkeratosis (EHK).Methods Clinical data were collected from two families with EHK.Peripheral blood was isolated from the probands and unaffected family members in the families as well as from 50 healthy controls.PCR was performed to amplify the encoding exons and flanking intron regions of KRT1 and KRT10 genes followed by direct DNA sequencing.Results Two mutations in the KRT10 gene,including a heterozygous acceptor splice site mutation in intron 4 (c.1030-2 A＞G) and a heterozygous missense mutation c.467 G＞A,were identified in the probands of both families,but absent in the unaffected family members or healthy controls.ConclusionThe splice site mutation c.1030-2 A＞G and missense mutation c.467 G＞A might be responsible for the phenotype of EHK in the two families.

Objective To observe the effects of standardized three stage rehabilitation treatment on the neu- rological deficit scores (NDS) and ADL performance of ischemic stroke patients.Methods A total of 164 ischemic stroke patients were recruited and randomly divided into a rehabilitation group and a control group.The neurological function and ADL performance of the patients were assessed by using NDS and Modified Barthcl Index (MBI) at the admission,at the end of 1st,3rd and 6th months post stroke.Results No significant differences were found be- tween the rehabilitative and the control groups with regard to NDS and MBI at admission.The NDS demonstrated a decreasing tendency,while the MBI score an increasing tendency in both groups.In the control group,significant difference of NDS was found between admission and the end of 1st month as well as between the end of the 1 st and the 3rd months.In rehabilitation group,significant difference was revealed between all the time points with regard to NDS and MBI scores.At the end of the 1st,3rd and 6th months,the MBI scores of the rehabilitation group were signifi- cantly higher than those of the control group,indicating that the ADL performance of those treated with standardized three-stage rehabilitation protocol was improved quicker than those without the protocol.Conclusion Standardized three-stage rehabilitation treatment could improve the neurological function and ADL performance of the ischemic stroke patients.

OBJECTIVETo construct a cell model of 4.1R gene knockout in murine macrophage cell line RAW264.7 using CRISPR/Cas9 technique.

METHODSThree high?grade small?guide RNAs (sgRNAs) that could specifically identify 4.1R gene were synthesized and inserted into lentiCRISPRv2 plasmid. RAW264.7 cells were infected with sgRNA?Cas9 lentivirus from 293T cells transfected with the recombinant sgRNA?lentiCRISPRv2 plasmid, and the positive cells were screened using puromycin and the monoclonal cells were obtained. The expression of 4.1R protein in the monoclonal cells was measured by Western blotting, and the mutation site was confirmed by sequence analysis. Result A 4.1R gene knockout RAW264.7 cell line was obtained, which showed a 19?bp deletion mutation in the 4.1R gene sequence and obviously enhanced proliferation.

CONCLUSIONWe successfully constructed a 4.1R gene knockout macrophage cell line using CRISPR/Cas9 technique, which may facilitate further investigation of the function of 4.1R in macrophages.

BACKGROUNDHuman piebaldism is a rare autosomal dominant condition characterized by congenital white forelock and depigmented patches of skin, typically on the forehead, anterior trunk and extremities. Mutations in the KIT gene have been proposed to be responsible for the underlying changes in this disorder. The aim of this study was to identify gene mutation in a Chinese family with piebaldism.

METHODSA Chinese family with piebaldism presenting with white forelock and large depigmented skin macules on the abdomen, arms and legs was collected. DNA was isolated from peripheral blood of the family members. The encoding exons with flanking intron regions of the KIT gene were analyzed by polymerase chain reactions (PCR) and direct DNA sequencing. Besides, DNA extracted from 100 ethnically matched population individuals was as controls.

RESULTSA heterozygous missense mutation c.2590T > C was identified in the patients of the family. This mutation converted a serine residue to proline (p.Ser864Pro). The mutation was not found in their unaffected family members or normal controls.

CONCLUSIONA novel missense mutation c.2590 T > C was found and it might play a significant role in the piebaldism phenotype in the family.

Child ; Humans ; Male ; Mutation, Missense ; Piebaldism ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; physiology

Objective:To analyze clinical and genetic characteristics of a family with familial generalized lentiginosis, and to identify the causative gene mutation.Methods:Clinical characteristics and inherited pattern were analyzed in a family with familial generalized lentiginosis. Peripheral blood samples were obtained from the proband, his affected father and healthy mother, and genomic DNA was extracted. PCR was performed to amplify all exons and their flanking sequences of the SASH1 gene, followed by DNA sequencing. The proband′s mother and 100 unrelated healthy controls served as controls to determine the mutation site. Previous literature and gene mutation databases were searched to rule out the possibility that the SASH1 gene mutations were single nucleotide polymorphisms, and to determine whether it was a known mutation.Results:A 4-generation family consisting of 17 members was investigated, and there were 9 patients in the family, including 7 males and 2 females. Patients existed in each generation, and the disease was inherited in an autosomal dominant manner in this family. Gene sequencing revealed a heterozygous duplication mutation c.49_54dupCCCGAG in exon 1 of the SASH1 gene in the proband and his father. This mutation was not found in his mother or healthy controls, and had not been reported in previous literature or gene mutation databases.Conclusion:The heterozygous duplication mutation c.49_54dupCCCGAG in the SASH1 gene is a pathogenic mutation for the clinical manifestations of familial generalized lentiginosis in this family.

BACKGROUNDIt has been found that cardiac protection afforded by ischemic preconditioning (IPC) is significantly reduced in the senescent myocardium. ADAMTS-1 (a disintesrin and metalloprotease with thrombospondin type 1 motifs) has been shown to inhibit angiogenesis in a variety of in vitro and in vivo assays. The aim of this study was to investigate the age-associated differences in ADAMTS-1 protein expression in rat myocardium after ischemic preconditioning.

METHODSSixty-four young (4 months) and old (24 months) male Sprague-Dawley rats were randomly assigned to an IPC group (40 rats) or a sham group (rats). A model of delayed IPC was induced and rats were sacrificed and myocardial samples were harvested from the ischemic-reperfused region for immunohistochemical detection of ADAMTS-1 at serial time points after IPC. A model of myocardial infarction was produced by ligation of the left anterior descending coronary artery in additional sets of young and old rats after sham or IPC procedures, then age-associated myocardial infarction survival after IPC was calculated.

RESULTSADAMTS-1 expression increased significantly in old rats compared to young rats (P < 0.05). The mean densities of ADAMTS-1 protein at 0, 6, 12, and 24 hours in young-IPC group after IPC were 0.05 ± 0.01, 0.13 ± 0.03, 0.16 ± 0.04, and 0.12 ± 0.03 vs. 0.07 ± 0.03, 0.20 ± 0.03, 0.24 ± 0.05, and 0.21 ± 0.04 in old-IPC group. IPC resulted in diminished survival rates (5/35 vs. 6/14, old-IPC group vs. old-sham group, P < 0.05), reduced left ventricular fractional shortening ((13.9 ± 2.8)% vs. (18.3 ± 2.3)%, P < 0.05) and increased the myocardial infarction size ((37.9 ± 3.2)% vs. (32.8 ± 5.1)%, P < 0.05) in the older rats.

CONCLUSIONSCardioprotection with IPC is attenuated in the older heart. ADAMTS-1 expression induced by IPC is greater in old rats. Over-expression of anti-angiogenic factors might be a potential mechanism behind reduced protection after IPC associated with aging.

ADAM Proteins ; metabolism ; ADAMTS1 Protein ; Aging ; metabolism ; physiology ; Animals ; Immunohistochemistry ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Infarction ; metabolism ; pathology ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

Objective To observe the effect of ischemia preconditioning (IPC) on the expression of pro-angiogenic VEGF, PDGF and anti-angiogenic ADAMTS-1, and arteriogenesis. Methods Rat heart IPC model was made by 4 circles of occluding the LAD for 6 min followed by 6 min of reperfusion. The expression of VEGF, PDGF-B and ADAMTS-1 in the ischemic area was examined with immunohistochemistry at 6, 12 and 24 h after IPC. IPC plus myocardial infarction model was induced by LAD ligation 24 h after IPC, 14 days later, the anti-SM-α-actin antibody was used to detect the mature neovascularization in the border of the infracted area. Results VEGF, PDGF-B and ADAMTS-1 were significantly upregulated in the ischemic area in IPC group compared with the control group ( P < 0. 05 ).Density of mature arteries was also significantly increased in IPC plus MI group than that in MI group (P <0. 05). Conclusion IPC promoted the formation of mature new arteries which may be modulated by upregulating VEGF, PDGF-B, and ADAMTS-1 expressions.

OBJECTIVETo determine whether serum homocysteine and folate levels are correlated with the occurrence of neonatal asphyxia and to study the effects of gender and gestational age on serum homocysteine and folate levels.

METHODSThirty-five neonates with mild asphyxia (19 males and 16 females) and 40 normal neonates (control group,18 males and 22 females) were enrolled in this study. The asphyxia and the control groups consisted of 10 and 11 cases of preterm infants respectively. Serum homocysteine levels were measured using ELASA. Serum folate levels were measured using radioimmunity assay.

RESULTSSerum homocysteine level (14.66+/-2.61 micromol/L vs 7.55+/-0.50 mumol/L; P<0.05) was significantly higher and serum folate level (2.47+/-0.24 ng/mL vs 3.28+/-0.28 ng/mL; P<0.05) was significantly lower in the asphyxia group than that in the control group. There were no significant differences in serum levels of homocysteine and folate between males and females either in the asphyxia group or the control group. The asphyxiated neonates born at premature showed increased serum homocysteine level compared with the full-term neonates with asphyxia (21.25+/-5.01 micromol/L vs 12.34+/-2. 01 micromol/L; P<0.05).

CONCLUSIONSThe increased serum homocysteine level and decreased serum folate level are correlated with the occurrence of neonatal asphyxia. Serum homocysteine and folate levels are not associated with the gender. A more significantly increased serum homocysteine level may be found in asphyxiated neonates born at premature.

Asphyxia Neonatorum ; blood ; Female ; Folic Acid ; blood ; Homocysteine ; blood ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Sex Characteristics

OBJECTIVETo investigate the effect of Qingre Quyu Granule (清热祛瘀颗粒, QRQYG) on stabilizing vulnerable plaques in apolipoprotein E (ApoE) deficient mice.

METHODSSeventy-two male ApoE deficient mice were given a high-fat diet from 6 weeks of age. At the 16th week, all the mice were randomized into 3 groups: the QRQYG group, the simvastatin group, and the control group. Sixteen weeks after administration of 0.9 g/kg QRQYG, 3 mg/kg simvastatin or 10 mg/kg sodium chloride per day to the respective groups, the animals were euthanized. The pathological morphologic changes in the vulnerable plaques were evaluated, the matrix metalloprotease-9 (MMP-9) expression was measured by immunohistofluorescence, the soluble intercellular adhesion molecule 1 (ICAM-1) was determined by ELISA, the nuclear factor kappaB (NF-κB) subunit p65 was measured by quantitative RT-PCR, and, finally, thrombospondin-1 (TSP-1) was determined by the immunohistochemical method.

RESULTSThe plaque cross-sectional area in the brachiocephalic artery (23.7%, P<0.01), the lipid core of the plaque (43.1%±3.1%), and the number of buried fibrotic caps of the plaque were significantly decreased in the QRQYG group compared to the control group (both P<0.01); furthermore, the thickness of the fibrotic cap of the plaque increased and the intra-plaque hemorrhage of the plaque decreased. The serum soluble ICAM-1 (27.1±5.1 μg/mL), the protein expression of MMP-9 and TSP-1 and the p65 mRNA expression increased in the QRQYG group in comparison with the control group (P<0.05 or P<0.01).

CONCLUSIONQRQYG could stabilize the vulnerable plaque through inhibition of the inflammatory response.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; pathology ; Brachiocephalic Trunk ; drug effects ; enzymology ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Knockout ; NF-kappa B ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Simvastatin ; pharmacology ; Sodium Chloride ; pharmacology ; Thrombospondin 1 ; metabolism