AIM: To investigate the effect of 4.25%peritoneal dialysis solution (PDS) on CD40 expression in rat peritoneal mesothelial cells so as to reveal the potential mechanisms by which CD40-CD40 ligand (CD40L) interaction may be involved in the inflammation of peritoneal membrane. METHODS: Rat peritoneal mesothelial cells (MC) were harvested from the peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with 4.25%PDS or 4.25%PDS+IFN-? was detected by RT-PCR and FACS analyses. After activation of CD40 on MC with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. RESULTS: MC cultured in vitro expressed CD40 constitutively. 4.25%PDS markedly up-regulated the expression of CD40 mRNA and its protein. The expression of CD40 mRNA and its protein following stimulation with 4.25%PDS+IFN-? was significantly higher than 4.25%PDS alone. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with CD40mAb.CONCLUSIONS: MC functionally express CD40. The up-regulated CD40 expression on MC following stimulation with 4.25%PDS may play an important role in local peritoneal defense mechanisms and may be involved in the chronic inflammatory process of the peritoneum.

OBJECTIVETo optimize the extraction procedure of essential oil from H. cordata using the SFE-CO2 and analyze the chemical composition of the essential oil.

METHODThe extraction procedure of essential oil from fresh H. cordata was optimized with the orthogonal experiment. Essential oil of fresh H. cordata was analysed by GC-MS.

RESULTThe optimize preparative procedure was as follow: essential oil of H. cordata was extracted at a temperature of 35 degrees C, pressure of 15,000 kPa for 20 min. 38 chemical components were identified and the relative contents were quantified.

CONCLUSIONThe optimum preparative procedure is reliable and can guarantee the quality of essential oil.

Aldehydes ; analysis ; chemistry ; Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Freeze Drying ; Gas Chromatography-Mass Spectrometry ; methods ; Houttuynia ; chemistry ; Ketones ; analysis ; chemistry ; Oils, Volatile ; analysis ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Pressure ; Temperature

OBJECTIVETo study the chemical constituents of the roots of Euphorbia soongarica (Xinjiang origin).

METHODCompounds were isolated and purified by repeated normal column chromatography, preparative thin layer chromatography and Sephadex LH - 20 chromatography. The chemical structures were elucidated by NMR and MS spectra.

RESULTSTen chemical constituents were isolated from the n-BuOH fraction and identified as ellagic acid (1) , 3, 3'-di-O-methylellagic acid (2) , 3, 3'-di-O-methylellagic acid-4'-O-alpha-D-arabinfuranoside (3), 3, 3'-di-O-methylellagic acid-4'-O-beta-D-xylopyranoside (4), 3, 3'-di-O-methylellagic acid-4-O-beta-D-glucopyranoside (5), 3, 3', 4'-tri-O-methylellagic acid (6), 3-O-methylellagic acid-4'-O-beta-D-xylopyranoside (7), 3, 3', 4-tri-O-methylellagic acid-4'-O-beta-D-glucopyranoside (8), brevifolin (9) and ethyl brevifolin carboxylate (10).

CONCLUSIONAll of above compounds were obtained from this plant for the first time.

Benzopyrans ; chemistry ; isolation & purification ; Ellagic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Euphorbia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

The isolate GN52 of Riemerrella anatipestifer was passaged on the Martin Medium successively according to the optimum condition. The experiments included Gram staining, biochemical test, drug sensitivity test and animal experiments were carried out on the bacteria of 3rd, 11th, 21st, 31st, 41st, 51st and 61st generations. It indicated that the bacterial morphs, biochemical character, drug resistance of the strain had no obvious change, but the virulence showed a trend of reduction.

A new species of genus shewarella Shewanellade decolorations S12, was isolated from activated sludge of a textile-printing waste-water treatment plant. In the anaerobic condition, S12 could conserve energy for growth by using Fe3 + as the terminal electron acceptor. At the optimal condition of pH8, temperature 30℃, ferric citrate 800mg/L, sodium lactate 2g/ L, yeast extract 0. 5g/ L , the cell growth increased with the raise of the amount of the Fe3+ reduction in 8k The effect of different carbon soucres, nitrogen sources, pH values and growth temperatures on the anaerobic Fe3 + reduction of Shewanella decolorations S12 was investigated. LB was favorable for Fe3 + reduction. Glucose and sodium lactate also were favorable for Fe3+ reduction. The cell growth and Fe3 + reduction increased with the raise of the amount of the yeast extract from 0 to 4g/L The amounts of the sodium lactate of 6g/ L and ferric citrate of 800mg/L were suitable for strain S12 growth and Fe3+ reduction. In the optimum initial pH value range of 6 -8 for Fe3+ reduction, strain S12 growth increased with the raise of the pH val- ue. Strain S12 could growth and reduce Fe3+ at the temperature range of 20 -40℃. The best temperature for strain S12 growth and Fe3 + reduction was 301.

Objective To investigate the relationship between recurrent spontaneous abortion (RSA) and the level of interferon-γ(IFN-γ) and interleukin-4 (IL-4) in peripheral blood and gingival crevicular fluid of patients with chronic periodontitis.Methods Fifty pregnant women with recurrent spontaneous abortion(gestational age: 5-28 weeks) and 40pregnant women (gestational age: 39-41 weeks)were included and their periodontal status was examined.The level of IFN-γ and IL-4 in gingival crevicular fluid and peripheral blood was detected by enzyme-linked immunosorbent assay.Results Nine RSA women and 5 in control group had periodontitis.The prevalence of periodontitis in RSA group was 18％ (9/50) and 13％ (5/40) in the control group(P ＜0.05).The probing depth,attachment loss and bleeding index in RSA group were higher than those in the control group(P ＜0.05).The level of IFN-γwas (52.98 ± 17.56) ng/L in gingival crevicular fluid of RSA group and (25.25 ± 7.93) ng/L in control group(P ＜ 0.01).The level of IL-4 was (15.43 ± 1.77) ng/L in gingival crevicular fluid of RSA group and (19.62 ± 4.04) ng/L in control group (P ＜0.01).The content of IFN-γwas (27.79 ± 3.59) ng/L in peripheral blood of RSA group and (18.39 ±2.65) ng/L in control group(P ＜0.05).The content of IL-4 was (15.88 ± 0.95) ng/L in peripheral blood of RSA group and (22.98 ± 4.30) ng/L in control group (P ＜ 0.001).The content of cytokines in gingival crevicular fluid and peripheral blood in the same group was positively related (r ＞ 0.8,P ＜ 0.001).Conclusions Pregnant women of RSA have a higher chance of getting periodontitis.The change in the level of cytokines in periodontal tissue could result in excursion of maternal intrauterine immune environment to T-helper 1 (Th1).Periodontitis was related to RSA.

OBJECTIVEInterleukin-12 receptor beta1 (IL-12 Rbeta1) deficiency is a rare primary immunodeficiency (PID) characterized by selective susceptibility to weakly virulent organisms, including Mycobacterium bovis, BCG, non-tuberculous environmental mycobacteria and non-typhoidal salmonellosis. The present study was conducted to identify the mutation type and to analyze clinical phenotype.

METHODSBased on the typical clinical manifestations and immunologic tests in this case, a varieties of PIDs were excluded and IL-12Rbeta1 deficiency was suspected. IL-12Rbeta1 chain expressed on Epstein-Barr virus-transformed lymphoblastoid B cell lines were detected by flow cytometric assay. The IL-12Rbeta1 gene sequences of the patient and her parents were analyzed by PCR-directed sequencing. The IL-12Rbeta1 gene sequences of the patient's younger brother also had been analyzed prenatally and after birth.

RESULTSAfter inoculating BCG, the patient suffered from multiple BCG infectious lymphadenitis. There was no detectable IL-12Rbeta1 on the Epstein-Barr virus-transformed lymphoblastoid B cell lines from the patient, while only mild expression on the cell line from her mother. Sequencing analysis by using sense and antisense primers separately, a novel IL-12Rbeta1 gene mutation was found in the patient which was homozygous single nucleotide substitution, a nonsense mutation with nucleotide substitution of C to T at position 853 (853C-->T) in exon 9 leading the glutamate at position 285 to the stop codon mutation (Q285X). The parents were carriers of the mutated IL-12Rbeta1 gene. But her younger brother has normal IL-12Rbeta1 gene.

CONCLUSIONThe novel IL-12Rbeta1 gene mutation is responsible for BCG infection in this case and genetic analysis is useful in carrier detection and prenatal diagnosis is feasible when the mother had a baby with identified IL-12Rbeta1 gene mutation before.

BCG Vaccine ; adverse effects ; Base Sequence ; Exons ; Female ; Humans ; Infant ; Molecular Sequence Data ; Mutation ; Mycobacterium bovis ; Phenotype ; Receptors, Interleukin-12 ; deficiency ; genetics ; Severe Combined Immunodeficiency ; complications ; genetics ; Tuberculosis ; complications

OBJECTIVETo investigate diagnostic histopathology and ultrastructure features of primary pulmonary cryptococcosis (PC).

METHODSClinical data and pathologic findings of 27 cases of PC were retrospectively reviewed, light and electron microscopic evaluations and histochemistry stain studies were performed.

RESULTSThe specimens consisted of 2 fine-needle aspiration lung biopsies and 25 cases of open lung biopsies. Cryptococcosis granuloma formation was identifiable by histopathological examination in 25 of 27 cases, with gum-like lesion and fungi in the remaining 2 cases. The detection rates of cryptococcus neoformans (CN) by mucicarmine (MC), periodic acid-Schiff (PAS), alcian blue (AB) and Grocott methenamine-silver (GMS) were 87.0% (20/23), 100% (27/27), 66.7% (18/27), and 100% (23/23) respectively. Under the electron microscope, most CN had a simple structure with a few organelles. The capsule was seen in all organisms. A percentage of the organisms showed nuclei, nucleoli, mitochondria and vacuoles. The detection rate of CN by EM was 91.7% (11/12).

CONCLUSIONSThe clinical manifestation and imaging of PC are nonspecific for PC. Lung biopsy is the major diagnostic modality. The detection rate by electron microscopy was quite high. Therefore, a correct diagnosis of pulmonary cyrptococcosis should rely on the combination of histopathological evaluation, histochemistry staining and/or electron microscopic examination.

Adult ; Aged ; Biopsy, Fine-Needle ; Cryptococcosis ; microbiology ; pathology ; Cryptococcus neoformans ; isolation & purification ; ultrastructure ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Lung ; pathology ; ultrastructure ; Lung Diseases, Fungal ; classification ; microbiology ; pathology ; Male ; Microscopy, Electron ; Middle Aged ; Retrospective Studies

BACKGROUNDEndothelial dysfunction is thought to be critical events in the pathogenesis of Alzheimer's disease (AD). Endothelial progenitor cells (EPCs) have provided insight into maintaining and repairing endothelial function. To study the relation between EPCs and AD, we explored the number of circulating EPCs in patients with AD.

METHODSA total of 104 patients were recruited from both the outpatients and inpatients of the geriatric neurology department at General Hospital, Tianjin Medical University. Consecutive patients with newly diagnosed AD (n = 30), patients with vascular dementia (VaD, n = 34), and healthy elderly control subjects with normal cognition (n = 40) were enrolled after matching for age, gender, body mass index, medical history, current medication and Mini Mental State Examination. Middle cerebral artery flow velocity was examined with transcranial Doppler. Endothelial function was evaluated according to the level of EPCs, and peripheral blood EPCs was counted by flow cytometry.

RESULTSThere were no significant statistical differences of clinical data in AD, VaD and control groups (P > 0.05). The patients with AD showed decreased CD34-positive (CD34(+)) or CD133-positive (CD133(+)) levels compared to the control subjects, but there were no significant statistical differences in patients with AD. The patients with AD had significantly lower CD34(+)CD133(+) EPCs (CD34 and CD133 double positive endothelial progenitor cells) than the control subjects (P < 0.05). In the patients with AD, a lower CD34(+)CD133(+) EPCs count was independently associated with a lower Mini-Mental State Examination score (r = 0.514,P = 0.004). Patients with VaD also showed a significant decrease in CD34(+)CD133(+) EPCs levels, but this was not evidently associated with the Mini-Mental State Examination score. The changes of middle cerebral artery flow velocity were similar between AD and VaD. Middle cerebral artery flow velocity was decreased in the AD and VaD groups and significantly lower than the normal control group (P < 0.01). There was no significant difference of the blood flow velocity between the AD and VaD patients (P > 0.05).

CONCLUSIONSThe results provided evidence that patients with AD have reduced circulating EPCs. Endothelial function is impaired in patients with AD and vascular factors have a role in the pathogenesis of AD. CD34(+)CD133(+) EPCs may be a novel biomarker of AD dementia.

AC133 Antigen ; Aged ; Alzheimer Disease ; metabolism ; pathology ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Dementia, Vascular ; metabolism ; pathology ; Endothelial Cells ; cytology ; metabolism ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Peptides ; metabolism ; Stem Cells ; cytology ; metabolism

OBJECTIVETo investigate the clinicopathological features and the immunohistochemical phenotype of perianal Paget's disease (PPD) associated with internal anorectal adenocarcinoma, with emphasis on the histogenesis of Paget's cells.

METHODSThe clinical and pathologic features of three cases of PPD with rectal adenocarcinoma were investigated. Periodic-acid-Schiff (PAS), alcian-blue and mucicarmine staining with and without diastase digestion were performed. The immunohistochemical study was performed on selected sections by a panel of antibodies including carcinoembryonic antigen (CEA), CK7, CK8, CK10/13, CK20 and gross cystic disease fluid protein 15 (GCDFP15).

RESULTSAll three cases occurred in middle to old age male patients complaining of anal bleeding. Digital physical examination revealed ulcerated or cauliflower-like masses in the anus just distal to the dentate line. Perianal skin erythematous patches were found in two cases, and small discrete granules in one case. Histologically, the anorectal neoplasm was either a moderately or poorly differentiated adenocarcinoma. Two types of Paget's cells were noted, namely the classical type characterized by a polygonal shape with vesicular nuclei and abundant pale cytoplasm, and the signet ring type characterized by eccentrically displaced nucleus. Both the rectal adenocarcinoma cells and Paget's cells showed strong positivity for PAS, AB and mucicarmine, which were resistant to the diastase digestion. Immunohistochemically, they were both positive for CEA, CK7, CK8 and CK20, but negative for CK10/13 and GCDFP15.

CONCLUSIONSThe CK20(+)-GCDFP15(-) type Paget's cells in PPD were derived from the direct intraepithelial Pagetoid spread of anorectal adenocarcinomas. PPD was more frequently associated with internal carcinomas than any other type of extramammary Paget's disease. It is recommended that clinicians should carefully examine the anus or rectum in the presence of PPD to ascertain if it is associated with an internal carcinoma.

Aged ; Aged, 80 and over ; Apolipoproteins ; Apolipoproteins D ; Carrier Proteins ; analysis ; Diagnosis, Differential ; Glycoproteins ; analysis ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; analysis ; Keratin-20 ; Male ; Membrane Transport Proteins ; Middle Aged ; Paget Disease, Extramammary ; chemistry ; diagnosis ; pathology ; Rectal Neoplasms ; chemistry ; diagnosis ; pathology