[Correlation of vasogenic white matter lesions and retinal vascular network parameters].

Objective: To investigate the correlation of vasogenic white matter lesions with retinal vascular network parameters using fully automatic retinal image analysis of fundus photographs. Methods: A total of 106 patients with cerebral small vessel disease who were hospitalized in Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University during March and October 2015, and were able to undertake cerebral MRI and fundus photography in a sitting position were included. They were divided into two groups (mild or moderate-severe) according to the Fazekas scores of periventricular white matter lesions and deep white matter lesions shown by MRI. The clinical data and retinal vascular network parameters were compared between mild and moderate-severe groups. Results: According to the severity of periventricular white matter lesions, Logistic regression analysis showed that after adjusting baseline information, decreased asymmetry index of artery (OR=1.71, 95%CI 1.02-2.88, P<0.05)was associated with periventricular white matter lesions. As for deep white matter lesions, Logistic regression analysis showed that after adjusting baseline information, decreased central retinal artery equivalent(OR=5.19, 95%CI 1.06-25.44, P<0.05), decreased asymmetry index of artery (OR=2.96, 95%CI 1.42-6.17, P<0.05), decreased asymmetry index of venule (OR=2.99, 95%CI 1.48-6.02, P<0.05) and increased central retinal vein equivalent (OR=0.14, 95%CI 0.03-0.67, P<0.05) were associated with deep white matter lesions. Conclusions: White matter lesions of different places could be contributed to different pathological process. Therefore, the early diagnosis and observation of them are applicable to different retinal vascular network parameters.

The role of CD4+ lymphocytes in the susceptibility of mice to stress-induced reactivation of experimental colitis.

Idiopathic inflammatory bowel disease is a chronic relapsing condition. The role of stress in causing relapses of inflammatory bowel disease remains controversial. We now show that colitis induced in mice by dinitrobenzenesulfonic acid (DNBS) resolves by 6 weeks, but can subsequently be reactivated by stress plus a sub-threshold dose of DNBS, but not by DNBS alone. Stress reduced colonic mucin and increased colon permeability. Susceptibility to reactivation by stress required CD4+ lymphocytes and could be adoptively transferred. We conclude that stress reactivates experimental colitis by facilitating entry of luminal contents that activate previously sensitized CD4 cells in the colon.

Phenotypic expression of non-Hodgkin's lymphomas in China.

Monoclonal antibodies were used to label malignant lymphomas obtained from 57 patients. On the basis of morphologic criteria, 18 lymphomas were the B-cell type, 10 were the T-cell type, and 6 were histiocytic; for 23 the type could not be determined. After monoclonal antibody labeling, 18 lymphomas of B-cell lineage were confirmed, 16 of the T-cell type were demonstrated, 6 were true histiocytic, and 17 were the null cell (non-T, non-B) type. Of the 16 lymphomas of T-cell lineage, 6 were lymphoblastic and 10 were the peripheral type. The percentages of cell types in the non-Hodgkin's lymphomas were as follows: B-cell, 31.5%; T-cell, 28%; null cell, 29%; and histiocytic, 10%. Of the 16 lymphomas of T-cell origin, 15 belonged to helper T-cell subsets (Leu1+, Leu4+, and Leu3a+), and the la marker was positive in all 16. Of the 18 B-cell lymphomas, 14 were kappa-positive and 4 were lambda-positive. Eleven were both B1- and kappa-positive, and 1 was kappa-positive but B1-negative. In the 4 cases that were lambda-positive, 2 were both lambda- and B1-positive. The results indicate that Leu4, Leu2a, Leu3a, and B1 are the most important markers to differentiate T-cell and B-cell lymphomas for pathologic classification. The findings also show a higher percentage of T-cell neoplasm in China as compared to that in Western countries.

Putative full-length clones of the genomic DNA segments of subterranean clover stunt virus and identification of the segment coding for the viral coat protein.

Subterranean clover stunt disease is an economically important aphid-borne virus disease affecting certain pasture and grain legumes in Australia. The virus associated with the disease, subterranean clover stunt virus (SCSV), was previously found to be representative of a new type of single-stranded DNA virus. Analysis of the virion DNA and restriction mapping of double-stranded cDNA synthesized from virion DNA suggested that SCSV has a segmented genome composed of 3 or 4 different species of circular ssDNA each of about 850-880 nucleotides. To further investigate the complexity of the SCSV genome, we have isolated the replicative form DNA from infected pea and from it prepared putative full-length clones representing the SCSV genome segments. Analysis of these clones by restriction mapping indicated that clones representing at least 4 distinct genomic segments were obtained. This method is thus suitable for generating an extensive genomic library of novel ssDNA viruses containing multiple genome segments such as SCSV and banana bunchy top virus. The N-terminal amino acid sequence and amino acid composition of the coat protein of SCSV were determined. Comparison of the amino acid sequence with partial DNA sequence data, and the distinctly different restriction maps obtained for the full-length clones suggested that only one of these clones contained the coat protein gene. The results confirmed that SCSV has a functionally divided genome composed of several distinct ssDNA circles each of about 1 kb.

Nicotine and gastric ulcers in stress.

Chronic nicotine treatment worsens stomach mucosal damage by cold (4 degrees C) and restraint (stress): it dose- and time-dependently intensifies stress-evoked gastric glandular ulceration, mast cell degranulation and motility. Nicotine 50 micrograms/ml drinking water, given ad libitum to female Sprague-Dawley rats for 10 days, increases the sensitivity of the isolated stomach strip to acetylcholine-induced contractions; atropine abolishes this action. The isolated anococcygeus muscle from nicotine-treated male rats shows increased sensitivity to noradrenaline-induced contractions, but not to those by acetylcholine. Hexamethonium or atropine pretreatment antagonises stress-induced gastric effects in nicotine-drinking rats. Muscarinic M1- and M2-, but not M3-, receptor block (by pirenzepine, AF-DX 116BS and HHSiD, respectively) inhibits stress ulcer formation in female rats. Although tobacco smoking has been reported to increase free radical formation, mucosal xanthine oxidase which initiates free radical formation is uninfluenced by nicotine; antagonising this enzyme (by allopurinol) or hydroxyl free radical scavenging (by dimethylsulfoxide) does not lessen the effect of nicotine on stress-evoked ulceration. The findings suggest that chronic nicotine treatment produces partial ganglionic blockade of the vagal nerve which leads to muscarinic receptor supersensitivity. This phenomenon contributes significantly to the ulcer-worsening mechanism; muscarinic M1- and M2-receptors appear to be involved. The gastric ulcer-aggravating effect of nicotine in stressed rats appears not to be due to increased free radical formation.

Effects of nicotine on activity and stress-induced gastric ulcers in rats.

Nicotine is known to influence locomotor activity. The alkaloid also intensifies gastric ulcer formation in stressed rats. The effects of nicotine on locomotor activity in relation to gastric lesions induced by restraint at 4 degrees C for 2 h (stress) were, therefore, studied. Ten-day treatment with nicotine 25 or 50 micrograms/ml drinking water potentiated stress-evoked ulceration and mast cell degranulation. These same doses of nicotine increased vertical motor activity; only the higher dose of the alkaloid enhanced horizontal movements. Phenobarbitone (12.5, 25, or 50 mg/kg, SC) dose dependently reduced vertical activity, as well as stress-induced gastric ulceration and mucosal mast cell degranulation. The drug also lessened the potentiating effects of nicotine on motor activity and stress-evoked gastric lesion formation. It is concluded that the ability of chronic nicotine treatment to intensify stress-induced gastric ulceration most likely owes part of its action to a mechanism evoking increased activity, which possibly reflects an influence on the CNS, as well as to enhancement of mast cell degranulation in the stomach glandular mucosa.

Effects of chronic nitric oxide synthase inhibition on TNB-induced colitis in rats.

Nitric oxide (NO) synthesis is increased in ulcerative colitis, but the role of NO in colitis is poorly understood. The present study employed Nw-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, in rats to evaluate the effect of NO on 2,4,6-trinitrobenzenesulphonic acid (TNB)-induced colitis. L-NAME solutions were placed in subcutaneous, osmotic mini-pumps which continuously released L-NAME at 0.042, 0.208, 0.417, or 1.667 mg kg-1 h-1. L-NAME dose-dependently enhanced lesions in TNB-induced colitis. The two higher doses of L-NAME significantly increased colonic mucosal damage, although there was slight, nonsignificant reduced lesion formation with the lowest dose of L-NAME. 0.042 mg kg-1 h-1. A single dose of L-NAME at 100 mg kg-1 subcutaneously injected daily in TNB-treated rats also increased lesions, and these ulcerogenic actions of L-NAME were reversed by L-arginine but not by D-arginine (both at 500 mg kg-1, s.c.). Only the highest dose of L-NAME (mini-pump) significantly depressed myeloperoxidase (MPO) activity. Faecal occult bleeding showed a close relationship with severity of colitis. These findings suggest that there may exist a balance between NO protective and aggressive effects. In TNB-induced colitis, antagonism of endogenous NO generation was intensified, whereas slight inhibition of NO synthesis reduced lesions. Variations in responses, related to timing or dose changes in L-NAME, may reflect the differences in inducible vs constitutive NO synthase isoforms.

An analysis of 24 patients with IgA deposition at the BMZ.

A study of 24 patients with IgA deposition at the BMZ of the skin showed that five conditions could be recognized: 1) linear IgA bullous dermatosis in adults (LAD, 7 cases); 2) linear IgA and IgG bullous dermatosis in adults (LAGD, 10 cases); 3) chronic bullous disease of childhood (CBDC, 3 cases); 4) dermatitis herpetiformis (DH, 1 case), and 5) systemic lupus erythematosus (SLE, 3 cases). Histopathologically, 5 of 7 patients with LAD were similar to the DH group, but 7 of 10 patients with LAGD were similar to the BP group. Half the patients with LAD and LAGD had oral lesions, and most of them had excellent responses to dapsone and Tripterygium Wilfordii, but the patients with CBDC did not respond to these treatments. In the patients with LAD and LAGD, the positivity rates of IgA anti-BMZ antibodies examined by indirect immunofluorescence (IIF) on intact skin and NaCl split skin were 41% and 64%, respectively. The heterogeneity of the histopathologic pictures of LAD and LAGD, the incidence of DH, and the value of using NaCl split skin for IIF are discussed.

Expression of proliferating cell nuclear antigen in cutaneous malignant lymphomas and lymphoid infiltrates.

Paraffin-embedded tissue of skin biopsy specimens taken retrospectively from 24 patients with cutaneous malignant lymphomas (CML) and 8 patients with cutaneous lymphoid infiltrates (CLI) and other dermatoses were studied retrospectively with PC10 immunostaining. The results show a statistical significant difference among PC10 indices for cutaneous genuine histiocytic lymphoma (CGHL), cutaneous germinal center cell-derived lymphomas (CGCCL), cutaneous peripheral T-cell lymphomas (CPTL), non-mycosis fungoides (MF) and Sezary's syndrome (SS), and MF when compared with those for CLI. There is a linear relationship between PC10 index and square root of PC10 density, both of which seem to have a parallel relationship to the severity of malignancy in CML. The nuclear volume of the positive tumor cell or lymphocyte with PC10 immunostaining may be also useful in differentiating CML from CLI.

DNA flow cytometry from paraffin-embedded tissue in diagnosis of cutaneous malignant lymphomas and pseudolymphomas.

DNA content and cell cycle distributions in paraffin-embedded blocks of 111 skin biopsy specimens of 70 patients with cutaneous malignant lymphomas (CML) and 41 patients with cutaneous pseudolymphomas (CPL) including chronic actinic dermatitis (CAD) were estimated by DNA flow cytometry. A statistical significant difference between DNA indices (DIs) or proliferative indices (PIs) for CML including mycosis fungoides (MF) II, MF III, Sezary's syndrome (SS), cutaneous peripheral T-cell lymphomas other than MF and SS, cutaneous germinal center cell-derived lymphomas and cutaneous genuine histiocytic lymphoma from CPL. DIs were also helpful in differentiating MF I from CPL. There was a linear relationship between DIs and PIs, both of which had a parallel relationship to the degrees of malignancy and mortality of varieties of CML. The finding of aneuploidy is likely to be useful in differentiating CML from CPL. It is worth noting that DIs, PIs and proportions of aneuploidy in CAD were all higher than those of higher malignancy of CML. These data could not be considered as markers of malignancy.

AGNORS in skin lesions of cutaneous malignant lymphomas and pseudolymphomas.

Nucleolar organiser regions (NORs) are loops of DNA situated on the short arms of acrocentric chromosomes 13, 14, 15, 21, and 22. They can be demonstrated in formalin-fixed paraffin-embedded sections by a one step silver technique; the resultant black structures are called AgNORs. The technique was used in 71 patients with cutaneous malignant lymphomas (CML) and 9 cutaneous pseudolymphomas (CPL). AgNORs in 200 nuclei were scored and the means, standard deviation and standard error of the means calculated. Counts were as follows: mycosis fungoides (MF) I (premycotic stage) 1.17 +/- 0.09, SEM = 0.01; MF II (infiltrating stage) 1.17 +/- 0.01, SEM = 0.02; MF III (tumor stage) 3.55 +/- 0.87, SEM = 0.43: cutaneous peripheral T cell lymphomas other than MF and Sezary's syndrome (SS) (CPTL) 2.55 +/- 1.11, SEM = 0.35; SS 1.52; cutaneous B cell lymphomas (CBCL) 2.18 +/- 0.18, SEM = 0.06; CPL 1.17 +/- 0.1, SEM = 0.03. There was a significant difference between counts for CML (MF III, CPTL, CBCL and SS) and CPL. Significant difference was also noted between scores for MF III and CBCL, and especially counts between tumor stage (MF III and CPTL) and pretumor stage (MF I and MF II) of cutaneous peripheral T cell lymphomas. Although the AgNOR technique is in the stage of research, it proves to be helpful in differentiating CML and CPL other than separating MF I and MF II from CPL.

Mucosal nitric oxide is not responsible for the hemodynamic changes induced by nicotine in rat stomachs.

It has been shown that chronic nicotine treatment decreases gastric mucosal blood flow (GMBF). The mechanism for this action is still not defined. In this study, nicotine treatment (5, 25 or 50 µg/ml drinking water) for 10 days dose dependently reduced the GMBF and volume of hemoglobin but increased ethanol-induced gastric damage. These effects were potentiated by N(ω)-nitro-l-arginine methyl ester (l-NAME), a nitric oxide (NO) synthase inhibitor. l-arginine but not the d-analog restored the actions of l-NAME, indicating that the selective action of l-NAME. However, the potentiating actions of l-NAME were significantly attenuated in the nicotine-pretreated rats. When the basal mucosal NO synthase (both iNOS and cNOS) activity and its second messenger cyclic GMP levels were measured, no difference was found between the nicotine and the non-nicotine groups. Furthermore, high dose of l-arginine could not reverse the action of nicotine. These findings suggest that the adverse action of chronic nicotine treatment on GMBF and lesion formation is probably mediated through a NO independent mechanism.

Expression of human hepatitis B virus surface antigen gene in transgenic tobacco.

Expression of Human hepatitis B virus surface antigen (HBsAg) gene in plant was reported for the first time. The recombinant plasmid pRoKII-HBsAg was constructed by inserting HBsAg gene into the downstream of CaMV 35S promoter of binary vector pRoKII and then introduced into Agrobacterium tumefaciens LBA4404. The kanamycin-resistant plants were obtained by Agrobacterium-mediated transformation system. It was shown that HBsAg gene was expressed in transgenic tobacco plants and their progenies by ELISA. The spherical particles of psi 22 nm is the leaf extract of transgenic tobacco were observed by immunosorbent electron microscopy.

[Use of baculovector for the expression of HBsAg gene in insect cells].

Based on the information of molecular biology of Autographa californica Nuclear Polyhedrosis virus (AcNPV), a recombinant transfer plasmid pAcMV was constructed by molecular procedures included using two synthetic localized probes, which provided an inserted position linked with BamHI sequences nearly at polyhedrin initiating ATG codon. Then an expression vector pAcMV-HBsAg was reconstructed, it contained HBsAg gene from subclone pYPSS-1 derived from adwserotype of HBV. The recombinant virus containing HBsAg gene was isolated and purified through 3 cycles plaques and hybridization experiment after cotransfection of Spodoptera frugiperda cells with DNA of pAcMV-HBsAg and AcNPV. The expression of HBsAg gene in S. frugiperda cells infected with recombinant virus AcRV-HBsAg was identified by ELISA as haemagglutination tests. The yield of HBsAg excreted from S. frugiperda cells (an appropriate density usually between 1-2 X 10(6) cells/ml) after 48-72 h infected with AcRV-HBsAg was 4-8 mg/L. HBsAg harvested from the infected culture medium was shown immunoelectromicroscopy to be composed of spherical particles of about 22 nm diameter. Using this purified HBsAg, Bal b/c mice was immunized, the titer of anti-HBsAg serum measured measured by RIA was similar to that of purified HBsAg from human blood. Stable recombinant virus was isolated and could be shown to replicate in corn borer (Ostrinia nubilalis) larvae. All of these results can be expected that this expression vector system will be commercially developed to its fullest potential for diagnosis and vaccine HBsAg.