Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

OBJECTIVETo investigate the roles of nitric oxide (NO) and eNOS in the pathogenesis of vasovagal syncope (VVS).

METHODSFourteen children with VVS (group A), 10 children with syncope other than vasovagal (group B) and 20 healthy volunteers (group C) were enrolled. Plasma NO levels in groups A and B were determined before and at the termination of the head-up tilt table test (HUT). The G894T polymorphism within the eNOS gene was determined in the three groups.

RESULTSPlasma NO levels in group A increased significantly when syncope attacked from 76.7+/-9.6 micromol/L (before HUT) to 90.0+/-11.4 micromol/L (P<0.05). After the syncope attack was improved, plasma NO level in group A was significantly reduced. There were no statistical differences in plasma NO levels before and after the HUT in group B. Determining the G894T polymorphism within the eNOS gene showed that group A was associated with a higher incidence of the GT gene type as compared to groups B and C (42.9% vs 10%; P<0.05).

CONCLUSIONSPlasma NO may be involved in the pathogenesis of VVS. The increased plasma NO level may be associated with the G894T polymorphism of the eNOS gene.

Child ; Humans ; Nitric Oxide ; blood ; physiology ; Nitric Oxide Synthase Type III ; genetics ; physiology ; Polymorphism, Genetic ; Syncope, Vasovagal ; etiology

The high molecular weight genomic DNA of yeast was extracted using three methods.Products were separated on agarose gel electrophoresis,quantified by spectrophotometer ND-1000 and restricted by EcoRⅠand MesⅠ.The result was shown that the genomic DNA extracted by modified benzyl chloride method was the best.The products of wild isolates supported it,too.This method was suitable for restriction of genomic DNA from yeast.

Objectives To investigate the expression and diagnostic value of 14-3-3 protein in brains of patients with sporadic Creutzfeldt-Jakob disease(sCJD).Methods 14-3-3 protein was immunohistochemically analyzed in tissue from the frontal lobe of 5 patients with sCJD and 4 non-CJD eases Using 14-3-3 ?and ?antibodies with reference to the results of KB,GFAP and PrP detection.Results The expressions of 14-3-3 protein in five brains of sCJD were more obviously,mostly in gray matters and astrocytes in three cases.The concentration was related to PrP deposition type,but not related to prion protein genotype.Except few expression of 14-3-3 protein in neurous of two cases of acute contusion,there were no expression in the other two cases in control group.Conclusions The expression of 14-3-3 protein in brain is useful to pathological diagnosis of CJD.

Limonin existed in citrus fruits has been shown to have anti-bacterial, anti-viral, anti-feedant, anti-nociceptive, anti-inflammatory activities and anti-carcinogenic activities. But the clinical use is limited by its low bioavailability. The aim of this study is to observe the absorption and secretion transport mechanisms of limonin in intestine which can pave the way for the further study and clinical use. The transport characteristics and mechanisms of limonin in rat were studied by in situ intestine perfusion and in vitro Caco-2 cells method. The intestinal absorption of limonin was probably via a facilitated diffusion pathway which was poor and without segment-selection. Verapamil and ketoconazole improved the absorption remarkably according to the result of in vitro Caco-2 cells study; however, probenecid had no significant effect on the absorption. The P-gp efflux and CYP3A4 metabolism were involved in the poor intestinal absorption and low bioavailability of limonin. The exploration of the intestinal absorption mechanism is crucial to the design of dosage form and clinical use of limonin.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Biological Availability ; Biological Transport ; drug effects ; Caco-2 Cells ; Cytochrome P-450 CYP3A ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Intestinal Absorption ; drug effects ; Ketoconazole ; pharmacology ; Limonins ; administration & dosage ; pharmacokinetics ; Male ; Perfusion ; Probenecid ; pharmacology ; Rats ; Verapamil ; pharmacology

Objective To explore a new 18F multifunctional synthesis module with two vessels to synthesize various kinds of 18F labeled radiopharmaceuticals for clinical utilities. Methods The module with two vessels contained five systems, capture of F ion, the first reaction vessel, the second reaction vessel, high performance liquid chromatography (HPLC) purification, and solid extraction. The two vessels were made of transparent glass and were used to prepare complex products by nucleophilic reaction or hydrolyzation. Results The complex compounds of 18F-ethyl-choline, 18F-N-succinimidyl-4-18 F-fluorobenzoate (SFB), and common compounds of 18F-fluorodeoxyglucose (FDG), 18F-3'-deoxy-3'-18 F-fluorothymidine (FLT) were synthesized by the new module. The uncorrected synthesis yield (EOS) of 18F-SFB, 18F-ethylcholine, and 18F-FDG were (28.2±1.9)% (n=5), (22.5±3.8)%(n=6), and (58.2±5.4)% (n=32), respectively. The corrected synthesis yield of 18F-FLT was (30.1 ±6.2)% ( n = 10). In addition,11C-N-methyl-N-( 1-methylpropyl )-1-( 2-corophenyl )-isoquinoline-3 -carboxamide ( PK11195 ) was also prepared by the module with the yield of (31.2 ± 2.5) % ( n = 3 ). Conclusions The new module with two vessels has been used for synthesis of many 18F compounds and it may have the potential to be used for other more 18F radiopharmaceuticals.

Background Posterior capsular opacification(PCO) is common complication after extrecapsular extract of cataract.Matrix metalloproteinases-3 (MMP-3) can degrade all the extracellular matrix except polyose.The gene therapy of PCO upon MMP-3 is the researching hot topic.Fibronectin ( FN ) is a degrade gelatin,so its expression can reflect the effect of MMP-3 on LECs indirectly. Objective The aim of this study was to construct MMP-3 eukaryotic recombination plasmid and transfect to lens epithelium cells(LECs) for the observation of MMP3 expression,and to explore the feasibility of gene therapy for after cataract. Methods Six fresh lenses were obtained from pigs.LECs were cultured using explant method.The eukaryotic expression vector pEGFP-N1-MMP-3 was reconstructed with MMP-3 and pEGFP-N1 plasmids.The accuracy of MMP-3 gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting pEGFP-N1-MMP-3 into LECs of pig,the expression of MMP-3 protein in the cells was indirectly observed by green fluorescent protein.The expression of FN in LECs was detected using Western blot. Results The result of double enzyme digestion was consistent with the base number of pEGFP-N1 plasmids and target fragment.By enlacing the result of DNA sequencing analysis with software,the resemblance of the DNA sequence of MMP-3 from recombination plasmid pEGFP-N1-MMP-3 and that of homo MMP-3 was 99.6％,indicating that the target fragment was inserted to pEGFP-N1 plasmids successfully.Green fluorescence for GFP was seen in the LECs in pEGFP-N1-MMP-3 transfected group,but absent response for GFP was in empty vector group.Western blot revealed that the relative expression level of FN in LECs was 0.666±0.008 in pEGFP-N1-MMP-3 trasfected group and 0.326 ±0.071 in empty vector group,with a significant difference between these two groups(P=0.000). Conclusions Eukaryotic recombination plasmid pEGFP-N1-MMP-3 is successfully constructed,and MMP-3 can be expressed in LECs after transfected.These results lay a foundation for the further research of MMP-3 gene therapy for PCO.

Background Leber hereditary optic neuropathy (LHON)is a mitochondrial DNA (mtDNA)hereditary disease,so it is significant to understand the influence of DNA mutation on the occurrence of LHON.Objective This survey was to evaluate the role of mtDNA mutation in the development of LHON.Methods This survey study was approved by the Ethic Committee of Wuhan General Hospital of Guangzhou Military Command and written informed consent was obtained from each subject before the relative medial examination.Seventy-two matrilineal relatives from a family with LHON were collected for a pedigree analysis and mutation screening.Regular eye examination was performed on 11 patients,13 mutant gene carriers and 49 individuals with normal phenotype,and the degree of visual damage was graded as follows: ＞0.3 was normal,0.1-0.3 was mild damage,＜0.05-0.1 was moderate damage,＜0.02-0.05 was severe damage and ＜0.01 was very severe damage.Clinical characteristics of LHON was evaluated.The periphery blood sample of 2-4 ml was collected from individuals to separate the mononuclear cells,and the mtDNA was extracted by modified high salt method.MtDNA was amplified by PCR and the mutation loci was sequenced.Results PCR amplification product sequencing of mutant gene showed that both G11778A and T14502C mutations were detected in 24 of 72 matrilineal relatives,but only 11 of 24 carriers developed LHON.No abnormal clinical findings were seen in the 13 carriers,showing a less 50％ penetrance in this family.There was no G11778A or/and T14502C mutation in the normal phenotype individuals of this family.The onset age for vision impairment in 11 affected matrilineal relatives varied from 8 to 50 years old,with the mean age of 24.36 years old,showing a significantly lower age than that of the 13 carriers (5-72 years old,mean 40.38 years old) (t =2.102,P=0.049).Conclusions This study suggests that the Gl1778A and T14502C mutation in mitochondrial DNA is one of causes in the development of LHON.The primary G11778A mutation together with T14502C mutation in mtDNA is a factor for the occurrence of LHON,hut it is not sufficient to the development of LHON.An effective “second hit” process will play an inducing role for LHON.

Objective:To compare a novel kind of anatomical locking plate versus proximal humeral internal locking system (PHILOS) in the treatment of proximal humerus fractures.Methods:A retrospective study was performed in the 35 patients with proximal humerus fracture who had been operatively treated at Department of Orthopedics, The Third Hospital of Peking University from January 2020 to June 2021. They were 13 males and 22 females, aged from 24 to 83 years (average, 56.7 years). Of them, 14 were fixated by the novel anatomical locking plate and 21 by PHILOS. The 2 groups were compared in terms of gender, age, fracture type, operation time, intraoperative blood loss, hospital stay, University of California (UCLA) shoulder scores and Disability of Arm Shoulder and Hand (DASH) scores for the upper limb dysfunction at one month after operation and at the last follow-up, and postoperative complications.Results:There was no significant difference in the preoperative general data between the novel plate group and the PHILOS group, showing comparability between them ( P>0.05). All the patients were followed up for 6 to 21 months (mean, 11.3 months). In the novel anatomical plate group, the operation time was (83.9±29.2) min, the intraoperative blood loss was (36.4±27.1) mL, the hospital stay was (2.3±1.1) d, the UCLA score and DASH score at one month after operation were (20.1±4.7) points and (55.5±19.1) points, the UCLA score and DASH score at the last follow-up were (28.5±4.6) points and (25.1±24.4) points, respectively; 2 patients developed complications after operation. In the PHILOS group, the operation time was (85.0±38.8) min, the intraoperative blood loss was (62.9±46.8) mL, the hospital stay was (2.4±0.9) d, the UCLA score and DASH score at one month after operation were (21.0±3.8) points and (49.6±23.7) points, and the UCLA score and DASH score at the last follow-up were (28.0±5.1) points and (19.1±17.3) points, respectively; 3 patients developed complications after operation. There was no significant difference in all the above items between the 2 groups (all P>0.05). Conclusion:In the treatment of proximal humerus fractures, the novel anatomical locking plate is a feasible internal fixator because it is comparable to PHILOS in fine efficacy, good fracture healing, and satisfactory functional recovery of the shoulder joint.

OBJECTIVETo investigate the expression and clinical significance of mismatch repair genes hMLH1 and hMSH2 in sporadic colorectal carcinoma tissues.

METHODSThe expression of hMLH1 and hMSH2 proteins was detected in the 63 sporadic colorectal carcinoma samples by immunohistochemical staining, including tumor tissue, adjacent tissue at 3 cm from the carcinoma, and normal tissue at 10 cm away from the tumor.

RESULTSThe positive rate of hMLH1 protein expression in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues was 95.2%, 85.7% and 81.0%, respectively. The positive rate of hMLH1 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.05). The positive rate of hMSH2 protein in the 63 normal colorectal tissues, adjacent tissues and sporadic colorectal carcinoma tissues were 76.2%, 66.7% and 52.4%, respectively. The positive rate of hMSH2 protein expression was significantly lower in the tumor than in normal colorectal tissues (P < 0.01). The positive rate of hMLH1 protein expression was significantly higher in the tumor tissue of patients aged younger than 60 years (100%) than that in patients ≥ 60 years (75.0%, P < 0.05). The positive rate of hMLH1 protein expression in the tumor tissue accompanied by lymphatic metastasis was 50.0%, significantly lower than that (93.3%) in tumors without lymphatic metastasis (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissue of patients aged younger than 60 years was 80.0%, significantly higher than that (43.8%) in the cases ≥ 60 years (P < 0.05). The positive rate of hMSH2 protein expression in the tumor tissues with invasion reaching to the intestinal serosa (61.5%) was significantly higher than that (37.5%) in the tumors invading to submucosa or muscular layer (P < 0.05). There was a positive correlation between the expressions of hMLH1 and hMSH2 proteins in the sporadic colorectal carcinomas.

CONCLUSIONThere is a certain loss of expression of hMLH1 and hMSH2 proteins in sporadic colorectal carcinoma, and is correlated with the age of patients, lymphatic metastasis and different depth of cancer invasion. HMLH1 and hMSH2 may be used as a useful laboratory marker in clinical judgement of occurrence and development of sporadic colorectal carcinoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; metabolism ; Neoplasm Invasiveness ; Nuclear Proteins ; metabolism ; Rectal Neoplasms ; metabolism ; pathology