Background: We found microRNA (miR)-1246 to be significantly differentially expressed between severe active alopecia areata (AA) patients and healthy individuals. Objective: To explore the role and mechanism of miR-1246 in severe AA. Methods: Expression of miR-1246, dual-specific tyrosine phosphorylation-regulated kinase 1A (DYRK1A), and nuclear factor of activated T cells 1c (NFATc1) in peripheral CD4+ T cells and in scalp tissues of patients were detected using RT-qPCR, Western blot, and immunohistochemistry assays. Peripheral CD4+ T cells from the AA patients were transfected with lentiviral vectors overexpressing miR-1246. RT-qPCR and Western blot analysis were used to measure mRNA or protein expression of retinoic-acid-receptor-related orphan nuclear receptor gamma (ROR-γt), interleukin (IL)-17, DYRK1A, NFATc1, and phosphorylated NFATc1. Flow cytometry was used to assay the CD4+ IL-17+ cells proportion. ELISA was used to measure cytokine levels. Results: miR-1246 levels decreased and DYRK1A and NFATc1 mRNA levels significantly increased in the peripheral CD4+ T cells and scalp tissues of severe active AA samples.NFATc1 protein expression was also significantly increased in the peripheral CD4+ T cells but not in the scalp tissues. NFATc1 positive cells were mainly distributed among infiltrating inflammatory cells around hair follicles. In peripheral CD4+ T cells of severe active AA, overexpression of miR-1246 resulted in significant downregulation of DYRK1A, NFATc1, ROR-γt, and IL-17 mRNA and phosphorylated NFATc1 protein, as well as a decrease in the CD4+ IL-17+ cells proportion and the IL-17F level. Conclusion miR-1246 can inhibit NFAT signaling and Th17 cell activation, which may be beneficial in the severe AA treatment.

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation

Adult ; Bone Plates ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Humeral Fractures ; diagnostic imaging ; physiopathology ; surgery ; Male ; Middle Aged ; Radiography

OBJECTIVETo prove analgesia and sedative effect of adjuvant anesthesia with magnetic auricular point-sticking on abdominal gynecological operation and its effect on postoperative recovery of body function.

METHODSNinety-two patients with abdominal gynecological operation were randomly divided into 3 groups. The auricular point-sticking group (APS group, n=31) was pasted and pressed by plasters with magnetic beads at bilateral Shenmen, Pizhixia (subcortex), Zigong (uterus) and Penqiang (pelvic cavity), etc. the night before operation. The placebo group (n=31) was pasted by plasters without magnetic beads. The blank group (n=30) was given no intervention. The mental and gastrointestinal functional changes before and 3 days after the operation were observed.

RESULTSAs compared with those in the control group and the blank group, the postoperative score of Self rating Anxiety Scale (SAS) was less (25.5 +/- 0.81 vs. 28.9 +/- 3.19, 28.3 +/- 2.36, both P < 0.01), with lower-dose of Innovar [(2.5 + 1.1) mL vs. (3.4 + 1.8) mL, (3.2 + 1.6) mL, both P < 0.05], earlier exsufflation after the operation [(34.2 + 12.1) h vs. (46.3 + 10.9) h, (43.2 + 14.8) h, both P < 0.01] and higher level serum of beta-endorphin before and after the operation in the APS group (all P < 0.05).

CONCLUSIONThe magnetic auricular point-sticking has sedative, analgesic and function-regulating effects on the abdominal gynecological operation.

Acupuncture Analgesia ; Acupuncture, Ear ; Adjuvants, Anesthesia ; administration & dosage ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; physiopathology ; therapy ; Postoperative Period ; Recovery of Function ; Young Adult

AIMTo investigate the influence of cytochrom P450 CYP2C9 polymorphism on the pharmacokinetics of tolbutamide.

METHODSAn oligonucleotide microarray was designed and fabricated to genotype the CYP2C9 accurately and quickly. 137 healthy volunteers were genotyped with the array to investigate the frequency of CYP2C9 functional SNPs. Moreover, 1 homozygous mutant, 9 heterozygous and 10 wild-genotypes subjects in the assay were selected randomly and sequenced directly. After orally taking tolbutamide, blood samples and urine samples were collected, and their pharmacokinetics was studied with HPLC.

RESULTSCYP2C9 *1/*3 were found in 9 of 137 volunteers, CYP2C9 *3/*3 in only one, others were all CYP2C9 *1/*1 wild types. CYP2C9 *2, CYP2C9 *4 and CYP2C9 *5 alleles were not detected. Direct sequencing of the purified PCR products of the heterozygotes, mutant homozygotes and ten wild type individuals gave a corresponding result to that genotyped by microarray. Pharmacokinetic outcome showed that the individuals with CYP2C9 *1/*3 or CYP2C9 *3/*3 had slower metabolic elimination of tolbutamide than those with CYP2C9 *1/*1.

CONCLUSIONCYP2C9 genetic polymorphism has a significant influence on the pharmacokinetics of tolbutamide. Pharmacogenomic study will be helpful in guiding rational and individualized medication. Key words: tolbutamide; cytochrom P450 CYP2C9; allele; single nucleotide polymorphism; genotyping

Aryl Hydrocarbon Hydroxylases ; genetics ; Cytochrome P-450 CYP2C9 ; Genotype ; Heterozygote ; Homozygote ; Humans ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Random Allocation ; Tolbutamide ; pharmacokinetics

This paper put forward a more accurate identification method for identification of Chinese materia medica (CMM), the systematic identification of Chinese materia medica (SICMM) , which might solve difficulties in CMM identification used the ordinary traditional ways. Concepts, mechanisms and methods of SICMM were systematically introduced and possibility was proved by experiments. The establishment of SICMM will solve problems in identification of Chinese materia medica not only in phenotypic characters like the mnorphous, microstructure, chemical constituents, but also further discovery evolution and classification of species, subspecies and population in medical plants. The establishment of SICMM will improve the development of identification of CMM and create a more extensive study space.
Drugs, Chinese Herbal ; Materia Medica

OBJECTIVETo investigate the regulatory effects of Shenfu Injection (SFI, ) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure (CHF).

METHODSForty-five healthy Wistar rats were randomized into three groups: sham, heart failure (model) and SFI group. The CHF was induced by left coronary artery ligation. Seven days after the surgical operation, animals in the sham group and the model group received saline (6.2 mL/kg/d), while animals in the SFI group received SFI (6.2 mL/kg d) intraperitoneally. Four weeks later, cardiac hemodynamic parameters were measured via the carotid route. The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS).

RESULTSRecording of hemodynamic parameters showed that left ventricular systolic pressure (LVSP), maximum rate of left ventricular pressure (+dp/dtmax) rise, and maximum rate of left ventricular pressure (-dp/dtmax) decrease, while the left ventricular end diastolic pressure (LVEDP) rose in the model group compared to those in the sham group (P <0.05). The results of the MALDI-TOF MS indicated that haptoglobin (HP), pentraxin 3 (PTX3) and alpha-1-antitrypsin were up-regulated, while serum albumin and 40S ribosomal protein were down-regulated in the model group (P <0.05). Compared with the model group, LVSP, +dp/dtmax and -dp/dtmax were higher, while LVEDP was lower in the SFI group (P<0.05). Expression levels of HP and PTX3 were lower than in the model group (P<0.05).

CONCLUSIONSFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF. The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure, and they could also be the serum protein targets of SFI.

Animals ; Blood Proteins ; metabolism ; C-Reactive Protein ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins ; metabolism ; Heart Failure ; blood ; complications ; drug therapy ; physiopathology ; Heart Function Tests ; Hemodynamics ; Hydrogen-Ion Concentration ; Imaging, Three-Dimensional ; Inflammation ; complications ; drug therapy ; Male ; Myocardial Ischemia ; blood ; complications ; drug therapy ; physiopathology ; Phytotherapy ; Proteome ; metabolism ; Rats, Wistar ; Serum Amyloid P-Component ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the activity of bilateral posterior cricoarytenoid muscle satellite cell after denervation or reinnervation with ansa cervicalis.

METHODSTwenty four dogs were randomly divided into 3 groups. The bilateral laryngeal recurrent nerves were cut in group one in all dogs. The bilateral laryngeal recurrent nerves were anastomosed with ansa cervicalis after incision in group two in all dogs. The dogs in group three were used as control. Nine weeks after surgery, the electromyography was used to test the regeneration of the nerve. The posterior cricoarytenoid muscles biopsy were collected. The expression of mRNA of Myogenin, Myf5, and Pax7 was assayed by realtime RT-PCR after total RNA isolation.

RESULTSTwo dogs died after surgery in incision and anastomose group. The electromyography suggested that the RLN of all dogs had denervated in the incision group and had reinnervated in the anastomose group after 9 weeks. Myogenin mRNA from RLN incision dogs PCA muscles had greater expression versus controls (Z = 1.42, P < 0.01) or anastomosed dogs (Z = 1.38, P < 0.01). Myf5 mRNA expression from RLN incision dogs PCA muscles had significant increase versus control dogs (Z = 1.66, P < 0.01) or anastomosed dogs (Z = 1.69, P < 0.01). Pax7 mRNA expression from RNL incision dogs had significant increase compared with control (Z = 1.66, P < 0.01) or anastomosed animals (Z = 1.42, P < 0.05). There was no significant difference in Myogenin (Z = 1.34, P > 0.05), Myf5 (Z = 0.54, P > 0.05) and Pax (Z = 0.54, P > 0.05) mRNA expression between controls and anastomosed animals.

CONCLUSIONSThe bilateral denervation of RLN cause significantly increasing in dog PCA muscle satellite cell proliferation and differentiation. The bilateral reinnervation of RLN cause PCA muscle satellite cell come back nonproliferative, quiescent state in dog.

Animals ; Cell Differentiation ; Cell Proliferation ; Dogs ; Laryngeal Muscles ; innervation ; Muscle Denervation ; Neck Muscles ; innervation ; Recurrent Laryngeal Nerve ; surgery ; Satellite Cells, Perineuronal ; cytology ; metabolism

Apoptosis is an important feature of atherosclerotic plaques, and it has both beneficial and detrimental effects depending on the cell type and plaque stage.Since apoptotic cells in advanced plaques launch proatherogenic inflammatory responses,efferocytosis is available to withstand atherosclerosis progression. Several efferocytosis sys-tems,composed of different phagocytic receptors,apoptotic ligands,and bridging molecules,can be identified.Im-paired efferocytosis pathway causes an overdue clearance of apoptotic cells and promotes secondary necrosis and the formation of necrotic core which leads to the instability and even rupture of plaques.

Purpose To discuss the clinical,histopathological characteristics,diagnosis,differential diagnosis and prognosis of hydroa vacciniforme-like lymphoproliferative disorder in children.Methods 6 cases of hydroa vacciniforme-like lymphoproliferative disorder were analyzed by molecular,histopathological and immunohistochemical testing.Clinical and follow-up information was obtained.The published relevant literatures were reviewed.Results 4 cases were boys,2 case were girls.All the patients presented with erythema and blisters with fever for 1 month to 4 years.Histopathologic examination showed an mild or moderate atypical lymphocytic infiltrate with angiotropism and angiocentricity,and scattered or dense lymphoid infiltration throughout the dermis and subcutaneous tissue.Blisters or necrosis could be seen in the epidermis.The atypical lymphocytes were positive for CD2,CD3,CDS,CD7,CD43,TIA-1,CD4 or CD8,and negative for CD20,Pax-5.Only one case showed positive stain for CD56.The average positive rate of Ki67 in tumor cells was 42.3％.Tumor cells positive for EBV encoded RNA (EBER) were detected in cutaneous infiltrates in 5 cases.Gene rearrangement of TCR was detected in 2 cases.5 patients were available for follow-up examination and 1 patient was dead.Conclusion Hydroa vacciniforme-like lymphoproliferative disorder is a rare disease mainly occuring in children.Chronic active EBV infection has been associated with this disease.It may be a spectrum in terms of its clinical course,and may be benign,borderline and malignant.Pathological diagnosis should be closely combined with clinical data.