OBJECTIVETo report novel mutations SEC23B gene in congenital dyserythropoietic anemia (CDA).

METHODSBy direct sequencing method, we sequenced CDAN1 and SEC23B genes in a Chinese CDA II patient, presented with chronic fatigue and dark urine, as well as his family members. Serum hepcidin was assayed by mass spectrometry.

RESULTSWe found a c.71G>A mutation and a c.74C> A mutation in the patient. In addition, a heterozygous c.55A>G mutation of HFE2 gene was found in some family members. The level of serum hepcidin of the patient was below the detection limit (<1 nmol/L).

CONCLUSIONContrary with what have been reported previously in the Europe, especially in the Italy, the gene mutations identified in this case was different and novel. The two novel mutations contribute to the diagnosis of CDAII and are the first report in East Asian CDAII patients.

Adolescent ; Adult ; Anemia, Dyserythropoietic, Congenital ; genetics ; Asian Continental Ancestry Group ; genetics ; GPI-Linked Proteins ; genetics ; Glycoproteins ; genetics ; Hepcidins ; blood ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Vesicular Transport Proteins ; genetics

OBJECTIVETo study the relation between the nitric-oxide synthase (NOS) expression and nitric oxide (NO) content in the skeletal muscles and the injury condition of soft tissue in the 3rd lumbar vertebrae syndrome model rats, and to observe the effect of acupotomology therapy.

METHODSOne hundred and twenty-eight adult SD rats were allocated to 4 groups randomly: normal group, model group, aminoguanidin group and acupotomology treatment group, 32 rats in each group. NOS expression, NO content and injury of the soft tissue in the 3rd lumbar vertebra were observed on the 1st, 3rd, 7th and 14th day after the acupotomology treatment and aminoguanidine intervention.

RESULTS1) Inducible NOS (iNos) activity and NO content in model group was significantly higher (F = 522.860, P < 0.01), in acupotomology group and aminoguanidine group was significantly lower than the model group (FiNOS = 28.894, P < 0.01), and iNOS activity and NO content in all groups was in competence with the condition of soft tissue injuries. 2) Endothelium NOS (eNOS) expression raised in model group and acupotomology group, and achieve peak on the 7th day. There was significant difference between the eNOS expression in acupotomology group and the model group (FeNOS = 3.454, P < 0.05). 3) The expression of neuron NOS (nNOS) in the model group, aminoguanidine group and acupotomology group had no significant (FnNOS = 0.962, P > 0.05).

CONCLUSIONAcupotomology treatment can restrain the development of high content NO, release the inflammatory reaction and injury condition, improve microcirculation, prevent the development of scar tissue of the injured soft tissue, and has significant recovering effectiveness in the soft tissue injured model rats.

Animals ; Disease Models, Animal ; Gene Expression Regulation, Enzymologic ; Guanidines ; therapeutic use ; Lumbar Vertebrae ; drug effects ; metabolism ; pathology ; surgery ; Male ; Muscle, Skeletal ; drug effects ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Time Factors

OBJECTIVETo observe the relationship between protein sythesis and cardiomyocyte viability in neonatal rats.

METHODSThe protein sythesis in neonatal rat cardiomyocytes was measured according to Brandford's method, the absorbance at 490 nm (A(490 nm)) of the cells was measured with MTT assay and the cell viability evaluated by the ratio of A(490 nm) to the total cell number.

RESULTSET-1 increased cardiomyocyte protein synthesis dose-dependently, and this effect was attenuated by the application of lacidipine and tetramethylpyrazines Higher doses of ET-1 resulted in lower A(490 nm)/total cell number ratio, which was further lowered by larcidipine and tetramethylpyrazine.

CONCLUSIONThe status of protein synthesis is not associated with the viability of neonatal rat cardiomyocytes.

Animals ; Animals, Newborn ; Calcium Channel Blockers ; pharmacology ; Cell Survival ; drug effects ; Cells, Cultured ; Dihydropyridines ; pharmacology ; Dose-Response Relationship, Drug ; Endothelin-1 ; pharmacology ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Protein Biosynthesis ; Pyrazines ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective: To investigate the expression of phosphorylated peroxisome proliferators-activated receptor γ (p-PPARγ) in the aging thoracic aorta of spontaneously hypertensive rat (SHR) and the inhibitory effect of rosiglitazone on the phosphorylation of PPARγ. Methods: 16, 32 and 64 week-old Wistar-Kyoto rats (WKY) and SHR were randomly and respectively divided into WKY, SHR and SHR+rosiglitazone group (9 in each group). The rats in SHR+rosiglitazone group were treated with rosiglitazone (5 mg/kg, intragastrically) for 56 d, whereas normal saline was applied in WKY and SHR groups. Systolic blood pressure (SBP) of rats was measured by tail cuff method. Histopathological damage of thoracic aorta was analyzed using Hematoxylin-Eosin (HE) staining. Immunohistochemical staining and western blot were performed to test the level of p-PPARγ protein in the thoracic aorta arising from each group. Results: The SBP in 16, 32 and 64 week-old SHR were significantly higher as compared with those in matched WKY rats (P < 0.05, respectively). HE staining showed increased content of smooth muscle cell, wrinkled lining endothelium and increased thickness of internal elastic lamina in the thoracic aorta of SHR. Immunohistochemical staining and western blot indicated that the levels of p-PPARγ in the thoracic aorta arising from SHR were obviously higher than those in the thoracic aorta arising from WKY rats (P < 0.05, respectively). Importantly, the high SBP, histopathological abnormalities of the thoracic aorta and elevated p-PPARγ expression were prominently abrogated by rosiglitazone treatment in SHR (P < 0.05, respectively). Furthermore, the SBP, histopathological abnormalities of the thoracic aorta and p-PPARγ expression were positively correlated with age in SHR (P < 0.05, respectively). Conclusions: The PPARγ phosphorylation was observed in the thoracic aorta of SHR and its expression was increased by the increase of age. Furthermore, rosiglitazone inhibited the PPARγ phosphorylation and suppressed vascular aging in SHR.

OBJECTIVETo systematically investigate the possible associations between G1165C and A145G polymorphism of β1-adrenoceptor (ADRB1) and resting heart rate (HRrest) in Northern Han Chinese.

METHODSHRrest of 700 healthy Northern Han Chinese were measured in the sitting position.SNPs were genotyped by the TaqMan assay.Genotypes were differentiated by analyzing the fluorescence levels of PCR products using an ABI Prism 7900HT Sequence Detector.

RESULTSHRrest was significantly lower in A145G AA carriers than in AG and GG carriers (all P < 0.01) . Multiple linear regression analysis showed that age, smoking habits, systolic blood pressure, triglyceride, serum creatinine and A145G polymorphism were associated with HRrest (P < 0.01) . A145G was significantly related with HRrest independent of other possible confounding variables, and the partial regression coefficient was 2.148 (P < 0.05) . After adjusting for other confounding factors, significant association between A145G and HRrest was only found in male subjects (P < 0.05) but not in female subjects (P > 0.05) .

CONCLUSIONThe A145G polymorphism of ADRB1 gene is associated with HRrest in Northern male Han Chinese.

Asian Continental Ancestry Group ; genetics ; Female ; Genotype ; Heart Rate ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, beta-1 ; genetics

Objectives: To explore the clinical experience for a bridge therapy of percutaneous balloon aortic valvuloplasty (PBAV) in treating the patients with severe aortic stenosis (AS). Methods: A total of 37 patients with severe AS who were not suitable for surgical valvular replacement received PBAV in our hospital from 2011-03 to 2017-03 were retrospectively studied. The patient's mean age was (74±12) years, their clinical and anatomical features, efficacy and safety of operation were observed and the outcomes were evaluated by follow-up study. Results: Patients presented the high surgical risk and worse cardiac function, 50% of them had bicuspid leaflet morphology with severe calcification [HU850=(856.0±658.2) mm3]. Balloon size was chosen by the intra-operative supra-annular diameters; at 7 days after operation, aortic valve orifice area (AVOA) was increased from (0.37±0.10) cm2to (0.87±1.10) cm2, the mean trans-aortic valve gradient pressure decreased form (55.1±22.9) mmHg to (44.8±17.8) mmHg, P<0.001 and LVEF elevated form(35.8±14.3)% to(41.0±12.2)%,P<0.001.There were 4 patients died in hospital,1 received permanent pacemaker and 1 developed severe aortic valve regurgitation. The patients were followed-up for (16.5±11.1)months after operation, 13/37 (35.1%) patients were in transition to surgical or transcatheter aortic valve replacement (TAVR). Conclusions: PBAV may have good early clinical efficacy in severe AS patients who were not suitable for surgical valvular replacement and TAVR; PBAV could be expected to become a bridge therapy, smaller supra-annular diameter was safe and effective for patients having bicuspid leaflet with severe calcification.

The study was aimed to establish a liquid chromatography-tandem mass spectrometric method for the determination of the duloxetine concentration in rat plasma, and compare the pharmacokinetics in normal and diabetes mellitus rat models. Diazepam was used as an internal standard. The separation was achieved on a Waters Xterra^® RP18 column (100 mm × 4.6 mm, 3.5 μm) with a mobile phase consisting of methanol -0.3% formic acid containing 5 mmol·L^-1 ammonium acetate (75:25) at the flow rate of 0.6 mL·min^-1. Electrospray ionization source was applied and operated in the positive multiple reaction monitoring mode. A good linearity of duloxetine was obtained in the concentration range of 10-5 000 ng·mL^-1. The rat models of diabetes mellitus were established by intraperitoneal injection of streptozotocin. The same dose of duloxetine (40 mg·kg^-1) was given by intragastric administration to the normal and diabetic rats. Blood samples were collected from the orbital venous plexus to determinate duloxetine concentration in the plasma. The pharmacokinetic parameters were calculated by DAS software. Statistical analysis was performed by SPSS software. The major pharma-cokinetic parameters of diabetes group were as follows: C_max was 1 185 ± 190.0 ng·mL^-1; AUC_0-∞ was 8 398 ± 1 835 ng·mL^-1·h; t_max was 1.6 ± 0.4 h; t_1/2z was 3.6 ± 0.9 h. The major pharmacokinetic parameters of normal group were as follows: C_max was 368.1 ± 40.7 ng·mL^-1; AUC_0-∞ was 4145 ± 640.1 ng·mL^-1·h; t_max was 1.6 ± 0.3 h; t_1/2z was 4.1 ± 0.8 h. The results of pharmacokinetic experiments suggest that the exposure amount of duloxetine in diabetic rats is twice higher than that in normal rats.

Many human genetic diseases, including Hutchinson-Gilford progeria syndrome (HGPS), are caused by single point mutations. HGPS is a rare disorder that causes premature aging and is usually caused by a de novo point mutation in the LMNA gene. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR/Cas9 nickase are highly efficient at inducing C to T base conversions in a programmable manner and can be used to generate animal disease models with single amino-acid substitutions. Here, we generated the first HGPS monkey model by delivering a BE mRNA and guide RNA (gRNA) targeting the LMNA gene via microinjection into monkey zygotes. Five out of six newborn monkeys carried the mutation specifically at the target site. HGPS monkeys expressed the toxic form of lamin A, progerin, and recapitulated the typical HGPS phenotypes including growth retardation, bone alterations, and vascular abnormalities. Thus, this monkey model genetically and clinically mimics HGPS in humans, demonstrating that the BE system can efficiently and accurately generate patient-specific disease models in non-human primates.
Animals ; Disease Models, Animal ; Female ; Gene Editing ; Humans ; Lamin Type A/metabolism* ; Macaca fascicularis ; Progeria/pathology*