Objective To evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P＜0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P＜0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P＞0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P＞0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P＜0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P＜0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P＞0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.

Background One of the features of the pathogenesis of primary angle-closure glaucoma(PACG) is anterior synechia of peripheral iris.Goniosynechialysis and combination of phacoemulsification and goniosynechialysis have been applied for the treatment of the disease recently,but the selection of operative types has great impact on clinical efficacy.Objective This study was to investigate the long-term efficacy of goniosynechialysis and combination of phacoemulsification and goniosynechialysis for the management of chronic PACG.Methods A non-randomized clinical controlled trial was designed.This clinical trial complied with Declaration of Helsinki and was approved by Medical Ethic Committee of Nanjing Medical University.Written informed consent was obtained from each patient.One hundred and ten eyes of 110 patients with chronic PACG were assigned to the goniosynechialysis group and combined operative group from March,2008 to February,2011 in Wuxi People's Hospital.180° goniosynechialysis was performed on 34 patients in the goniosynechialysis group,and phacoemulsifieation +intraocular lens (IOL) implantation + goniosynechialysis were carried out in 78 patients of the combination operative group.All the patients were followed-up for 2 years.Vision acuity,intraocular pressure (IOP),anterior chamber depth(ACD),unltrasound biomicroscopy and perimetry were recorded and compared between before and after operation.Results No significant difference was found in vision acuity between preoperation and postoperative 2 years in the goniosynechialysis group ([0.65 ± 0.15] vs.[0.45 ± 0.15]) (t =1.57,P＞0.05),but in the combination operative group,the vision acuity was significantly different between the before and after operation ([0.25±0.15] vs.[0.85 ±0.05]) (t =9.12,P＜0.001).The lOPs at 2 years after operation were (14.2±4.1) mmHgand(13.7±4.8) mmHg,respectively in the goniosynechialysis group and combination operative group and were significantly lower than(47.2 ±6.3) mmHg and(46.9±7.0) mmHg before operation(t =4.95,P＜0.001 ;t=5.03,P＜0.001).The ACD values in the goniosynechialysis group and combination operative group were(3.38±0.02)mm and (3.54±0.03) mm 2 years after operation,which were significantly increased in comparison with (1.33 ±0.24)mm and (1.56±0.37) mm before operation(t=7.65,P＜0.001;t=6.76,P＜0.001).Conclusions Both combination of phacomulsification with goniosynechialysis or 180° goniosynechialysis are effective for the treatment of PACG.Suitable operation should be alternated depending on the indicators of PACG patients.

Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.

To establish neonatal rat cardiac fibroblast inflammatory secretion model by using LPS 100 microg x L(-1) combined with ATP 5 mmol x L(-1), in order to study the inhibitory effect of quercetin on the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts, further investigate the effect of quercetin on the protein expression of p-NF-kappaB p65 (S276) and p-Akt (S473) by western blot, and discuss the inhibitory effect of quercetin on the inflammatory secretion of cardiac fibroblasts. According to the findings, quercetin with the concentrations between 51.74 micromol x L(-1) and 827.81 micromol x L(-1) had no significant effect on the activity of cardiac fibroblasts. Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of TNF-alpha and IL-1beta induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.01 or P < 0.05). Quercetin with the concentrations of 82.78, 41.39 micromol x L(-1) could notably inhibit the increase of IL-6 induced LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.05), without any notable effect of quercetin with the concentration of 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20. 70 micromol x L(-1) could notably inhibit the NF-kappaB p65 (S276) activation induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 15 min, with the most significant effect in 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of p-Akt(473) expression induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 240 min (P < 0.05). Therefore, this study believes that quercetin could attenuate the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts by inhibiting the activation of NF-kappaB p65 (S276) and Akt (473).
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Endomyocardial Fibrosis ; drug therapy ; genetics ; immunology ; Female ; Fibroblasts ; drug effects ; immunology ; Heart ; drug effects ; Humans ; Interleukin-6 ; genetics ; immunology ; Male ; Proto-Oncogene Proteins c-akt ; genetics ; immunology ; Quercetin ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; immunology

AIM AND METHODSThe parameters of low frequency stimulation (LFS) were altered systematically (frequencies of 1, 3 or 5 Hz; number of pulses of pulses of 300 or 900; and time lag after high frequency stimulation (HFS) of 20 or 100 min) and examined their effects on depotentiation (DP) of long-term potentiation (LTP) of synaptic transmission in CA1 neurons in hippocampal slices of rat.

RESULTSLTP could be induced by HFS (two trains of 100 Hz, 100 pulses, separated by 30 s) and be reversed to produce DP by a train of LFS of 900 pulses at 3 Hz given 20 min after HFS. DP induced by LFS could be blocked by NMDA receptor antagonist AP5 (50 micromol/L). And significantly reduced effect was observed for LFS at 1 Hz or 5 Hz, with smaller numbers of pulses or a longer time lag from LFS to HFS.

CONCLUSIONThe above results indicate that DP induced in CA1 neurons of rat hippocampal slices is strongly dependent on the parameters of LFS, and the process may be mediated through the NMDA receptor.

Animals ; Electric Stimulation ; methods ; Hippocampus ; cytology ; physiology ; In Vitro Techniques ; Long-Term Potentiation ; Male ; Neurons ; physiology ; Rats ; Rats, Sprague-Dawley ; Synapses ; physiology ; Synaptic Transmission ; physiology

Macrophages are involved in many important biological processes and membrane proteins are the key effector molecules for their functions. However, membrane proteins are difficult to analyze by 2-DE based method because of their intrinsic tendency to self-aggregate during the first dimension separation (IEF). To circumvent the obstacle hampering membrane protein analysis, we combined one-dimensional SDS-PAGE with capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this technique, we identified 458 GO annotated membrane proteins with extremely high confidence, including most known markers of peritoneal macrophages (e.g., CD11b, F4/80, CD14, CD18, CD86, CD44, CD16 and Toll-like receptor). Thirteen other CD antigens and 18 Ras-related small GTPase were also identified. In addition to those known macrophage membrane proteins, a significant number of novel proteins have also been identified. This research provides a valuable data set of macrophage membrane proteins, thus allowing for more comprehensive study of membrane proteins and a better understanding of the function mechanisms of macrophages in many biological processes.

Objective： This study was designed to investigate the effectiveness of natural and naphthyl imide-conjugated antisense oligonucleotide on inhibiting mdr1 gene expression and of overcome the problem of multidrug resistance in human epidemic carcinomata anti-adriamycin cells(KB-A-1). Methods： A novel naphthyl imide-conjugated mdr1 antisense oligonucleotide was synthesized by S-alkylation reaction. Compared with the natural one, the effectiveness of naphthyl imide-conjugated mdr1 antisense oligonucleotide on inhibiting the multidrug resistance was detected by MTT colometric assay and ELISA. Results： The abilities of antisense oligonucleotides to inhibit the cell growth of KB-A-1 mainly depended upon their targeted sequences selected. The inhibiting rates to KB-A-1 cells of antisense oligonucleotides (from Oligo Ⅰ to Oligo Ⅳ ) which bind the targeted sequences of the translation initiation site of mdr1 were 13.34％ ,14.32％ ,26 00％ ， and 25.37％ ， respectively. Among four oligonucleotides,the activity of Oligo Ⅲ to inhibit the growth of KB-A-1 cell was the highest (P<0.01). Compared with the unmodified one, the inhibition of antisense oligonucleotides to the growth of KB-A-1 cell was increased by conjugating with naphthyl imide. The ability of naphthyl imide-conjugated antisense oligonucleotide to resist serum-mediated nuclease was also increased as demonstrated by gel shift electrophoresis. The antisense oligonucleotide or its naphythyl imide conjugate could only inhibit the growth of KB-A-1 cells and had no effect on the growth of KB-3-1 cells. So it could be inferred that the inhibition of KB-A-1 cells was due to the repression of gene expression of mdr1 in KB-A-1 cells by antisense oligonucleotide or its naphthyl imide conjugate. ELISA showed that the P-glycoprotein expressions were more strongly inhibited by naphthyl imide-conjugated oligonucleotide than that by unmodified oligonucleotide. Conclusion: Inhibition of human tumor cell growth was due to the inhibition on mdr1 gene expression and the reversal of multidrug resistance by antisense oligonucleotide and its naphthyl imide conjugated. 1, 8-Naphthyl imide could improve the properties of natural oligonucleotide in inhibiting the multdrug resistance and resisting nuclease by covalently linkage at the end of oligonucleotide.

Objective To investigate the clinical efficacy of Zhu's "neck seven needles" in treating cervical spondylotic radiculopathy. Method Ninety-six patients with cervical spondylotic radiculopathy were randomized to two groups. The treatment group received Zhu's "neck seven needles" therapy and the control group, cervical vertebra traction therapy. Both groups were treated once every other day, for a total of 10 days. The symptoms were scored using the Short-form McGill Pain Questionnaire (SF-MPQ-2) before and after treatment. Result The SF-MPQ-2 score decreased significantly in both groups of patients after treatment compared with before. It decreased more in the treatment group than in the control group and the difference was statistically significant (P<0.01). The total efficacy rate was significantly higher in the treatment group than in the control group (P<0.01). Conclusion Zhu's "neck seven needles" is clinically more effective than conventional cervical vertebra traction in treating cervical spondylotic radiculopathy.

One pair of degenerate primer was designed according to conserved motifs of the psaB (A2 subunit of photosystem I) of Chlamydomonas reinhardtii, Chlamydomonas moewusii, Chlorella vulgaris and Mesostigma viride, and a total RNA of Dunaliella salina (D. salina) was extracted with TRIzol reagent. A cDNA fragment, about 1.8kb in length, from green algal D. salina was obtained through RT-PCR method. The resulting PCR product was cloned into T-vector and screened to determine its sequence. Homologous analysis of the deduced amino acid sequence was performed by BLAST and subsequeqtly compared with GenBank data. The obtained cDNA sequence was 1815 bp long, which encodes 605 amino acids (GenBank accession number: AY820754). The sequence shared high homologue with the following psaB: Chlamydomonas reinhardtii 92%, Chlamydomonas moewusii 91%, Chlorella vulgaris 86%, Mesostigma viride 85%, Physcomitrella patens subsp. Patens 85% and Nephroselmis olivacea 84%. It can be concluded that the cloned sequence is psaB cDNA fragment from D. salina.
Algal Proteins ; genetics ; Amino Acid Sequence ; Animals ; Chlamydomonas reinhardtii ; genetics ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Molecular Sequence Data ; Photosystem I Protein Complex ; genetics ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid

OBJECTIVETo optimize the acupuncture treatment programs for facial paralysis.

METHODSSixty-three patients of facial paralysis were randomly divided and treated according to the table of L9 (3)4 in orthogonal test. They were treated with different combined programs of 4 factors and 3 levels, including factor A (acupuncture op portunity), B (acupoints prescription), C (quantity of stimulus) and D (time of electroacupuncture). The change of facial nerve function score was observed to choose the best acupuncture treatment program for facial paralysis from factor A (acupuncture opportunity), B (acupoints prescription), C (quantity of stimulus), D (time of electroacupuncture) and their 3 levels in each factor.

RESULTSB (acupoints prescription) and D (time of electroacupuncture) were significant factors (P < 0.05), and B (acupoints prescription) was the most important influential factor. B3 (alternative use of two groups of acupoints) was the best one among the 3 levels of B (acupoints prescription), and D3 (electroacupuncture in disperse-dense wave for 30 min) was the best one of D (time of electroacupuncture).

CONCLUSIONTwo groups of acupoints alternatively used with electroacupuncture in disperse-dense wave for 30 min is the best treatment program for facial paralysis.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Facial Paralysis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult