[Objective] To observe NGF on cultured human retinal vascular endothelial cells (HRCEC) proliferation. [Methods] The MTT assay was used to analyze the impact of culture HRCEC on different factors (NGF concentration groups, NGF + K252a concentration groups, bFGF group, bFGF + K252a groups, the normal culture medium groups) in normal and hypoxic condition. [Results] With the increase of NGF concentration (20,50,100 ng/mL), HRCEC significantly increased (normal condition: 0.254±0.033,0.696±0.029, 1.136±0.051; hypoxic condition: 0.422±0.036, 0.798±0.044, 1.376±0.052, P< 0.05). Compared NGF + K252a group with the same concentration of NGF (100 ng/ml) group, HRCEC reduced (P<0.05), with increasing the concentration of K252a (50,100,200 nmol/L), the trend of HRCEC decreasing is become more significant (normal condition:0.864±0.067, 0.496±0.025, 0.202±0.078; hypoxic condition:K252a 1.042±0.047,0.700±0.065, 0.401±0.078, P<0.05). [Conclusion] NGF can promote the proliferation of HRCEC, the effect could be specifically blocked by TrkA inhibitor K252a.

OBJECTIVE:To study the influence of calcineurin gene polymorphism on the efficacy of cyclosporine A (CsA). METHODS:The blood samples of patients treated with CsA were collected. The trough blood concentration of CsA was detected by EMIT. The genotype of PPP3CA and PPP3CB was assayed by RFLP-PCR method. The expression of NFAT-regulated gene IL-2, IFN-γand GM-CSF were measured by RT-qPCR,which were used to define the index of indirect efficacy of CsA. The relationship of gene polymorphism with CsA efficacy was study by relationship analysis and multiple factor regression method,etc. RESULTS:A to-tal of 100 blood samples were collected. There was no significant correlation between the expression of CsA efficacy-related NFAT-reg-ulated gene GM-CSF and trough concentration of CsA(rGM-CSF=-0.04,P=0.238);the expression of IL-2 and IFN-γ were negatively correlated with trough concentration of CsA significantly(rIL-2=-0.384 3,P<0.001;rIFN-γ=-0.335 4,P<0.001). Using the average value of mRNA expression of IL-2 and IFN-γas indirect efficacy index,the polymorphism of PPP3CB rs3763679 site significantly in-fluenced the efficacy of CsA(P<0.05),but PPP3CA rs3804358 had no any effect on it(P>0.05). Stratified analysis showed that among patients with immune disease underwent renal transplantation,efficacy of patients with PPP3CB rs3763679 genovariation(TC+TT) were better than those with wild-type gene (CC)(P<0.05). After the efficacy was normalized by CsA trough concentration, multivariate analysis showed that normalized efficacy of CsA was negatively correlated with gender,PPP3CB rs3763679,lactate dehy-drogenase and creatinine significantly,but positively correlated with PPP3CA rs3804358,leucocyte count,usea nitrogen,glycerin trilaurate,etc. CONCLUSIONS:PPP3CB rs3763679 gene polymorphism influence the efficacy of CsA;among patients with immune disease underwent renal transplantation,efficacy of patients with PPP3CB rs3763679 TT+TC is better than that of CC type. At the same time,gender,PPP3CA rs3804358,leucocyte count,usea nitrogen,glycerin trilaurate and other factors all can influence the nor-malized efficacy of CsA to different extent. Multiple factors should be considered when using CsA.

To get the information about civil attitudes and main affecting factors of cadaver donation in Zhengzhou.Make stochastic visits to part of dwellers in Zhengzhou urban districts and then study the statistical data.The main affecting factors are traditional ideas and unsound legal system.So,in order to have a good effect,we should strengthen the dissemination of cadaver donation,renovate ideas,and strengthen relevant legislation for cadaver donation.

OBJECTIVE To investigate the effect of injection of β2-adrenergic receptor agonist clenbuterol into the infralimbic cortex(IL) on drug-seeking behavior triggered by conditioned cues. METHODS Adult male SD rats were trained to self-administer heroin under a FR1 schedule for consecutive 14 d,followed by 2-h extinction training. Cue-induced heroin seeking was measured for 2 h. Clenbuterol was microinjected bilaterally into the IL(8 ng/side)of rats 15 min prior to reinstatement test. Meanwhile,locomotor activity was detected 15 min after clenbuterol or artifial cerebrospinal fluid(mod?el group) was microinjected bilaterally into IL. Western blotting was used to detect the expression of phosphorylated cyclic AMP response element-binding protein(p-CREB)in the prelimbic cortex(PL), IL,nucleus accumbens core (NACc) and shell (NACsh) of rats immediately after reinstatement test. RESULTS After heroin administration training for 14 consecutive days,these animals exhibited reliable heroin self-administration,indicated by the increase in active nose poke responses and infusions. The rats that had received infusion of clenbuterol into the IL had significantly lower active pokes (8 ± 3)than those in model group(45±10)in cue-induced reinstatement(P<0.01),but there was no significant differ?ence between clenbuterol group and vehicle group in the locomotor activity. The expression of p-CREB in either IL or NACsh was significantly decreased in clenbuterol group compared with model group(P<0.01,P<0.05),but significantly increased in NACc(P<0.01). CONCLUSION Microinjection of clenb?uterol into the IL can attenuate the cue-induced reinstatement of heroin-seeking behavior in rats. The underlying mechanism might be related to the regulation of p-CREB expression in the NACc and NACsh.

OBJECTIVE:To establish the HPLC fingerprints for Huobahuagen tablets intermediate. METHODS:HPLC was per-formed on Inertsil ODS-4 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid (gradient elution) at the flow rate of 0.75 mL/min. The detection wavelength was set at 220 nm,and column temperature was 30 ℃. The sample size was 10 μL. Using wilforgine as reference,HPLC chromatograms of 10 batches of samples were determined. Common peak identification and similarity evaluation were performed by using Similarity Evaluation System for TCM Chromatographic Fingerprint (2004 A edi-tion). RESULTS:There were 25 common peaks in HPLC chromatograms of 10 batches of samples,and similarity degrees were higher than 0.9. After validated,HPLC chromatograms of 10 batches of samples were in good agreement with control fingerprints. CONCLUSIONS:The established fingerprint can provide reference for identification and quality evaluation of Huobahuagen tab-lets intermediate.

Rheumatoid arthritis (RA) is a chronic, inflammatory, autoimmune disorder that principally attacks flexible joints and synovia. The precise pathogenesis of RA remains unclear, and genetic factors probably play an important role in its etiology. In addition to genes from human leukocyte antigen (HLA) region, such as HLA-DRB, genes from non-HLA region, such as TIM-3, PTPN22, TRAF1/C5, STAT4, CCR5, PADI4 and FCGR2A may also contribute to its susceptibility. The advance in molecular genetics research on RA is reviewed here.
Arthritis, Rheumatoid ; genetics ; Exome ; Genetic Predisposition to Disease ; HLA-DRB1 Chains ; genetics ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Membrane Proteins ; genetics ; Protein Tyrosine Phosphatase, Non-Receptor Type 22 ; genetics ; Receptors, IgG ; genetics

OBJECTIVETo assess the association of copy number variations of SMN1, SMN2, NAIP, GTF2H2 and H4F5 genes with clinical classification of spinal muscular atrophy in children, and determine the copy number of the SMN gene among pregnant women. A carrier screening was also performed in Sichuan province.

METHODSThe copy number variations of the above genes among 53 confirmed SMA patients were determined with MLPA technique. The copy number variations were analyzed by the Fisher's exact test. Deletion of exon 7 in the SMN1 gene was screened with denaturing high performance liquid chromatography (DHPLC) for 427 pregnant women.

RESULTSAmong the 53 cases of type I, II, and III SMA patients, the rate of homozygous deletion of both exons 7 and 8 of the SMN1 gene were 100%, 94.44% and 87.50%, respectively, whereas those of homozygous deletion of exon 7 of SMN1 gene were 0, 5.56%, and 12.50%, respectively. The patients with 1, 2, 3, and 4 copies of exon 7 of the SMN2 gene were 11.32%, 67.92%, 13.21% and 7.55%, respectively. The patients with 0, 1, and 2 copies of exon 5 of NAIP gene were 11.32%, 62.26%, and 26.42%, respectively. No deletion was detected in GTF2H2 or H4F5 genes. The heterozygous loss rate of exon 7 in SMN gene in the pregnant women population of Sichuan region was approximately 2.11%.

CONCLUSIONCopy number variations of SMN2 and NAIP genes in patients are related to SMA clinical types (P < 0.05). In contrast, there was no relationship between SMA clinical types and deletion of exons 7 and 8 in the SMN1 gene (P > 0.05). Analysis of copy number change in SMN1 gene can assist SMA carrier screening. However, when the general population without SMA family history is screened for disease-causing genes, it should be noted that the type "2+0" carriers may affect the screening result, and the result should be interpreted with caution.

Adolescent ; Child ; Child, Preschool ; DNA Copy Number Variations ; Female ; Genetic Carrier Screening ; Humans ; Infant ; Male ; Neuronal Apoptosis-Inhibitory Protein ; genetics ; Spinal Muscular Atrophies of Childhood ; genetics ; Survival of Motor Neuron 1 Protein ; genetics

OBJECTIVETo analyze the mutation of COL9A2 gene and investigate the molecular pathogenesis of pathological myopia in a Han Chinese population.

METHODSMutation in the coding region of the COL9A2 gene was screened by Sanger sequencing in 200 subjects with pathological myopia and 200 normal controls. The detected variants were genotyped by SNaPshot method in another 200 myopic cases and 200 normal controls.

RESULTSSanger sequencing has failed to detect the reported D281fs frameshift mutation in the 200 cases. A novel variant, c.143G>C heterozygous missense mutation in exon 2, was identified in a myopic subject, and another novel variant, c.884G>A heterozygous missense mutation in exon 17, was found in another case. Neither was found in normal controls. One SNP (rs2228564) was detected in the coding region of the COL9A2 gene, but there was no significant difference in its allelic frequencies between the two groups (P> 0.05). Genotyping of the remainder 200 cases and 200 controls by SNaPshot method has found a c.143G>C in 1 case and c.884G>A in 2 cases, though no significant difference between the two groups was detected (P> 0.05).

CONCLUSIONThe D281fs frameshift mutation in the COL9A2 gene is not associated with pathological myopia in the studied Han Chinese population. Two novel mutations, c.143G>C in exon 2 and c.884G>A in exon 17 of the COL9A2 gene, may contribute to the development of pathological myopia.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Collagen Type IX ; genetics ; Frameshift Mutation ; Humans ; Myopia, Degenerative ; genetics ; Sequence Analysis, DNA

Objective To prepare an ophthalmic solution of dihydromyricetin(DMY)based on dipotassium glycyr-rhizinate(DG)nanomicelle solubilization(DG-DMY)and evaluate its effect on dry eyes of mice.Methods DG-DMY was prepared using the thin-film hydration method,and its micelle size,potential,encapsulation efficiency and storage sta-bility at room temperature were tested.The ocular safety of DG-DMY was tested on mice.Dry eye models were built in mice,which were divided into normal control group(normal mice without intervention),PBS control group(dry eye mouse models,intervened by PBS),HA treatment group[dry eye mouse models,intervened by 1 g·L-1 hyaluronic acid(HA)]and DG-DMY treatment group(dry eye mouse models,intervened by DG-DMY),with 10 mice in each group.The fluorescein sodium staining of corneal epithelium and surface tear secretion were recorded after 10 days of intervention.Morphological changes in corneal epithelium,corneal stroma and endothelial cells were monitored by hematoxylin & eosin staining.The enzyme-linked immunosorbent assay(ELISA)was adopted to measure the expression levels of interleukin-6(IL-6)and interleukin-1β(IL-1β).Results DG-DMY is a light yellow,transparent solution with a nanomicelle size of(208.8±3.9)nm,polydispersity index of 0.277,Zeta potential of-(17.6±1.42)mV,encapsulation efficiency of 76.72％,and drug loading efficiency of 10.21％.It is stable at room temperature(25℃)and storage temperature(4 ℃).The mouse studies showed that DG-DMY displayed good in vivo tolerance in mice eyes.The therapeutic results showed that mice in the PBS treatment group still had extensive corneal staining,mice in the HA treatment group had reduced corneal staining,and mice in the DG-DMY treatment group had almost no corneal staining.The tear secretion of mice in the normal control group,PBS control group,HA treatment group and DG-DMY treatment group was(5.15±0.47)mm,(2.26±0.41)mm,(4.02±0.53)mm,and(4.11±0.54)mm.The histopathological results showed that the corneal epithelium,loose collagen structure and basal layer were damaged in the PBS control group;the corneal histopathological injury of mice in the HA treatment group and DG-DMY treatment group were mitigated,with normal corneal epithelium,corneal stroma and endothelial tissues.ELISA results showed that the expression level of IL-6 in the normal control group,PBS control group,HA treatment group and DG-DMY treatment group was(22.98±0.69)ng·g-1,(108.1±6.06)ng·g-1,(56.79±4.87)ng·g-1 and(44.01±0.99)ng·g-1,respectively,and the expression level of IL-1β was(27.97±2.74)ng·g-1,(115.70±5.16)ng·g-1,(50.36±1.56)ng·g-1 and(42.21±1.46)ng·g-1,respectively.Compared with the HA treatment group,the expression levels of IL-6 and IL-1β in the cornea of mice in the DG-DMY treatment group were lower,and the differences were statistically significant(both P＜0.05).Conclusion DG-DMY nano-preparation successfully prepared in this study is verified to act on benzalkonium chloride-induced dry eye effectively and control the inflammatory response of dry eye mouse models by inhibiting the expressions of IL-6 and IL-1β with high safety.

The purpose of this research is to investigate the therapeutic effects of losartan potassium on 5/6 nephrectomy rats with chronic heart failure(CHF)and to explore the mechanism. 24 Rats were randomly divided into three groups namely sham group(Sham), pathology group(Nx)and losartan potassium group(Lst), respectively. CHF model in rats were induced by 5/6 nephrectomy. At the 7th week, rats of Lst group were given losartan potassium(50 mg/L)for consecutive 2 weeks. Then all rats were measured for hemodynamic parameters, cardiac index, creatinine, urea nitrogen in serum, and expressions of CD133, VEGFR2, Sox2, cleaved Caspase-3 and Bcl-2 in heart. Compared with Nx group, rats of Lst group improved cardiac and renal functions: decreased LVDP, LVEDP, cardiac index, creatinine, urea nitrogen and increased LVSP. Furthermore, losartan potassium up-regulated gene expression of CD133, VEGFR2, Sox2 and protein level of Bcl-2, and down-regulated cleaved Caspase-3 protein expression. Results suggest that losartan potassium can improve cardiac function of rats with CHF which may be correlated with mobilizing bone marrow stem cells, increasing endothelial progenitor cells(EPCs)level in heart, repairing endothelial function, and inhibiting myocardial apoptosis.