Lactobacillus paracasei PS23 reduced early-life stress abnormalities in maternal separation mouse model.

Maternal separation (MS) has been developed as a model for inducing stress and depression in studies using rodents. The concept of the gut-brain axis suggests that gut health is essential for brain health. Here, we present the effects of administration of a probiotic, Lactobacillus paracasei PS23 (PS23), to MS mice against psychological traits including anxiety and depression. The administration of live and heat-killed PS23 cells showed positive behavioural effects on MS animals, where exploratory tendencies and mobility were increased in behavioural tests, indicating reduced anxiety and depression compared to the negative control mice (P<0.05). Mice administered with both live and heat-killed PS23 cells also showed lower serum corticosterone levels accompanied by higher serum anti-inflammatory interleukin 10 (IL-10) levels, compared to MS separated mice (P<0.05), indicating a stress-elicited response affiliated with increased immunomodulatory properties. Assessment of neurotransmitters in the brain hippocampal region revealed that PS23 affected the concentrations of dopaminergic metabolites differently than the control, suggesting that PS23 may have improved MS-induced stress levels via neurotransmitter pathways, such as dopamine or other mechanisms not addressed in the current study. Our study illustrates the potential of a probiotic in reversing abnormalities induced by early life stress and could be an alternative for brain health along the gut-brain axis.

A thin-layer chromatography-bioautographic method for detecting dipeptidyl peptidase IV inhibitors in plants.

A thin-layer chromatography (TLC)-bioautographic method was developed with the aim to detect dipeptidyl peptidase IV (DPP IV) inhibitors from plant extracts. The basic principle of the method is that the enzyme (DPP IV) hydrolyzes substrate (Gly-Pro-p-nitroaniline) into p-nitroaniline (pNA), which diazotizes with sodium nitrite, and then reacts with N-(1-naphthyl) ethylenediamine dihydrochloride in turn to form a rose-red azo dye which provides a rose-red background on the TLC plates. The DPP IV inhibitors showed white spots on the background as they blocked enzymolysis of the substrate to produce pNA. The method was validated with respect to selectivity, sensitivity, linearity, precision, recovery, and stability after optimizing key parameters including plate type, time and temperature of incubation, concentration of substrate, enzyme and derivatization reagents, and absorption wavelength. The results showed good lineary within amounts over 0.01-0.1µg range for the positive control, diprotin A, with the coefficient of determination (r(2))=0.9668. The limits of detection (LOD) and quantification (LOQ) were 5 and 10ng, respectively. The recoveries ranged from 98.9% to 107.5%. The averages of the intra- and inter-plate reproducibility were in the range of 4.1-9.7% and 7.6-14.7%, respectively. Among the nine methanolic extracts of medicinal herbs screened for DPP IV inhibitors by the newly developed method, Peganum nigellastrum Bunge was found to have one white active spot, which was then isolated and identified as harmine. By spectrophotometric method, harmine hydrochloride was found to have DPP-IV inhibitory activity of 32.4% at 10mM comparing to that of 54.8% at 50µM for diprotin A.

Norisoboldine attenuates inflammatory pain via the adenosine A1 receptor.

BACKGROUND: Norisoboldine (NOR) is a benzylisoquinoline alkaloid isolated from Radix Linderae, a traditional Chinese medicine. Our previous studies have demonstrated that it produces anti-inflammatory and anti-rheumatoid arthritis effects. METHODS: The present study was undertaken to explore the analgesic effects of NOR and its potential mechanism in the formalin test and the acetic acid writhing test. RESULTS: Oral administration of NOR dose dependently attenuated the formalin-induced pain responses in the second phase, and reduced formalin-induced paw oedema. It also diminished acetic acid-induced writhing responses but had no effect on acute thermal pain in the hotplate test. The mechanistic studies suggested that the adenosine system, but not the opioid receptor system, is involved in NOR-induced antinociception. Naloxone, a non-selective opioid receptor antagonist, had no effect on NOR-induced analgesic action. However, caffeine (a non-selective adenosine receptor antagonist) completely reversed the analgesic effect of NOR in formalin-induced nociceptive responses in the second phase, and 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX, a selective adenosine A1 receptor antagonist) completely inhibited NOR-induced analgesia in both formalin-induced nociceptive responses and acetic acid-induced writhing responses. In addition, NOR reduced formalin-induced activation of extracellular signal-regulated kinase and calcium/calmodulin-dependent protein kinase II in the spinal cord, which is also blocked by DPCPX. Furthermore, NOR decreased forskolin-evoked cyclic adenosine monophosphate levels in mouse spinal cord neuronal cultures through the adenosine A1 receptor. CONCLUSION: Our data demonstrate that NOR produces the analgesic effect in inflammatory pain by a mechanism related to the adenosine system.

Therapeutic effect of norisoboldine, an alkaloid isolated from Radix Linderae, on collagen-induced arthritis in mice.

The alkaloid fraction of Radix Linderae, the main active component of this herb drug, has been proven to exhibit anti-inflammatory, analgesic and antimicrobial activities. The present study was undertaken to investigate the therapeutic potential of norisoboldine, the major isoquinoline alkaloid present in Radix Linderae, in collagen II -induced arthritis (CIA) of mice as well as the possible mechanisms. CIA was induced in mice by immunization with chicken type II collagen (II). After boosted on day 21, mice were treated with norisoboldine (10, 20, 40 mg/kg) for twenty consecutive days. The clinical scores, body weight changes and joint histopathology were evaluated. Norisoboldine treatment significantly alleviated the severity of the disease, based on the reduced clinical scores and elevated the lowered body weights of model mice. Meanwhile, this alkaloid dose-dependently reduced the infiltration of inflammatory cells, synovial hyperplasia and protected joint from destruction. Additionally, the serum level of anti-CII IgG and the CII-stimulated lymphocyte proliferation were remarkably decreased in the groups administered with norisoboldine. An assessment of Th1 function using the delayed-type hypersensitivity model confirmed that norisoboldine also significantly suppressed the enhanced T cell responses in vivo. These findings suggest that norisoboldine might be a potential therapeutic agent for rheumatoid arthritis, and it functions through protecting joint destruction as well as regulating the abnormal immune responses.

A new phenolic glycoside from the roots of Paeonia veitchii.

From the roots of Paeonia veitchii Lynch., a new phenolic glycoside, 2-O-[alpha-L-arabinopyranosyl-(1 --> 6)-beta-D-glucopyranoside]-benzaldehyde (1) was isolated together with seven known phenolic compounds. Their structures were elucidated by spectroscopic method.

Mycobacterium avium complex-associated hemophagocytic syndrome in systemic lupus erythematosus patient: report of one case.

Hemophagocytic syndrome (HPS) in systemic lupus erythematosus(SLE) patients has not commonly been reported. In this case study, we report the first case of Mycobacterium avium complex (MAC)-associated hemophagocytic syndrome in a patient with systemic lupus erythematosus (SLE). This SLE patient, a 15-year-old girl, had been on a high dose of prednisolone (> 0.5mg/kg/day) for more than 3 years. She presented with a spiking fever, hepatosplenomegaly, pancytopenia, hyperferritinemia and adult respiratory distress syndrome. Bone marrow examination revealed hemophagocytosis as well as non-caseating granulomatosis. There was no indication of SLE fare-up. She responded poorly to initial treatment with methyl-prednisolone, intravenous immumoglobulin, etoposide, and drugs for Mycobacterium tuberculosis including rifampin, ethambutol, isoniazid and pyramide. However, gastric lavage culture revealed MAC. Following treatment with clarithromycin, ciprofloxacin and amikacin, her condition gradually improved and she was discharged 3 months after admission. In SLE patients with pancytopenia and hyperferritinemia, MAC-associated HPS should be considered in the differential diagnosis.

Salvage of completely degloved finger with a posterior interosseous free flap.

The use of a free posterior interosseous skin flap should be considered in a single digit degloving injury, especially when replantation of the avulsed skin or the use of skin from the second toe, transferred as a composite-tissue flap, is not feasible. The flap is thin and pliable. It allows early mobilisation with good recovery of joint motion and attains protective sensation of the finger.

[Studies on constituents of the leaves of Lindera aggregata (Sims) Kosterm].

OBJECTIVE: To study the antibacterial and anti-inflammatory constituents of the leaves of Lindera aggrega. METHOD: Compounds were isolated by colum chromatography, and the structures were identified by spectroscopic methods. RESULT: Six compounds were isolated and identified as mixture of 6-Acetyllindenanolide B-1 and B-2(I), dehydrolindestrenolide (II), hydroxylinderstrenolide (III), linderalactone (IV), kameofero (V), beta-sitosterol (VI). CONCLUSION: These compounds were obtained from the leaves of Lindera aggregata for the first time.

Barrett's esophagus in a child with de Lange syndrome: report of one case.

Barrett's esophagus, a premalignant condition, is recognized as stratified squamous epithelium of the esophagus substituted by columnar epithelium. The risk factors for development of Barrett's esophagus include frequent gastroesophageal reflux, esophageal stricture, male sex and mental retardation, but there is no report of Barrett's esophagus in children with de Lange syndrome. We report a 7-year-old boy who was diagnosed as de Lange syndrome shortly after birth and had gastroesophageal reflux since early infancy. Upper gastrointestinal endoscopic examination revealed a cauliflower-like mass and a pink-red velvety mass over the lower third of the esophagus. Biopsy showed goblet cells metaplasia, confirming Barrett's esophagus. We suggest surveillance of Barrett's esophagus could be done ahead of schedule in children with long-standing gastroesophageal reflux or with de Lange syndrome.

[Isolation of a gene related to trichosanthin-induced apoptosis (GRETA)].

Disturbance of the apoptosis-related signaling pathway is regarded as one of the critical factors for tumorigenesis. Isolation of the genes involved in the process of apoptosis would be thereby helpful to explore the mechanism of tumor transformation and to develop novel therapeutic approaches. Here we report a gene fragment GRETA, the gene related to trichosanthin-induced apoptosis, isolated from a leukemia cell line U937 undergoing apoptosis induced by a plant protein Trichosanhin (TCS). A 293bp segment of GRETA was revealed to be 78.3% homologous to Bruton's tyrosine kinase at nucleic acid level. And Northern blot analysis showed that three messengers of RNA with the size of about 0.8-kb, 2.0-kb and 7.0-kb, respectively, were detected in TCS-untreated U937 cells when probed with GRETA, but there were only 0.8-kb and 2.0-kb transcripts appeared in apoptotic U937 cells. In addition, the abundance of each transcript changed apparently. The 0.8-kb transcript, for example, was the main band in Northern analysis in apoptotic U937 cells while was only detected marginally in TCS-untreated cells. These data suggested a possible relationship between the alternative splicing patterns of GRETA and the apoptosis.

Cytotoxic cembrenolides and steroids from the formosan soft coral Sarcophyton crassocaule.

Three new cytotoxic cembrenolide diterpenes, sarcocrassolide (1), crassolide (2), and 13-acetoxysarcocrassolide (3); a known cytotoxic cembrenolide, denticulatolide (4); and two cytotoxic steroids, (24S)-24-methylcholestane-3beta,5alpha,6beta-triol (5) and 24zeta-methylcholestane-3beta,5alpha,6beta,25-te traol-25-monoacetate (6), have been isolated from the Formosan soft coral Sarcophyton crassocaule. The structures of compounds 1-6 were determined by 1D and 2D spectral analysis.

Skin injury in the operating room.

Skin injury occurring in the operating room may occur as a result of electrical current, thermal injury, chemical irritation and mechanical stress. Between 1 December 1996 and 28 February 1997, 19 cases of skin injury from a total of 3657 operations were noted in our hospital. These injuries tend to be ignored because of their early spontaneous healing. We suggest that medical staff should pay more attention to this complication and prevent it from occurring.

The comparison of early fluid therapy in extensive flame burns between inhalation and noninhalation injuries.

Over the last half century, advances in treatment have changed the principal cause of death in burn patients from burn shock and wound sepsis to pulmonary sepsis, of which inhalation injury has always played a key role in morbidity and mortality. Even though Navar et al., Am. J. Surg. 1985;150:716-720 have noted that patients with inhalation injury had a mean fluid requirement of 5.8 ml/kg/% burn to achieve resuscitation from early burn shock, while patients without inhalation injury required only a mean fluid of 4.0 ml/kg/% burn, to achieve successful resuscitation in inhalation injured patients with minimum but adequate fluid has always been a challenge. To further define the difference of early fluid therapy between inhalation and noninhalation in extensively burned patients, a retrospective analysis was carried out in the Tri-Service General Hospital. Sixty-two flame burned patients (aged from 16 to 81 years-old with a mean age of 33.2+/-15.1 years: with burn size ranging from 30% to 98% TBSA with a mean burn size of 60.5+/-22.7%; 26 with inhalation injury; noninhalation 36) were reviewed during a 5-year period. The Parkland formula is the initial choice of fluid regimen with 4 ml/kg/% burn and the amount of replacement is monitored by urine output and is titrated to maintain urine output between 0.5 and 1.0 ml/kg/h. The mean amounts of fluid requirements of both inhalation and noninhalation burned patients were 3.1 +/- 1.0 and 2.3+/-0.8 ml/kg/% burn respectively (p < 0.05). Our study showed less fluid requirement for both inhalation and noninhalation injured patients in comparison with the Navar study and Parkland predictions in the first 24 h postburn. Furthermore, the inhalation injured patients definitely required volumes of fluid in excess of those required in noninhalation injured cases.

[Modulation of T-cell-mediated immune responses by trichosanthin via antigen processing and presentation].

It has been shown in our previous study that the lymphoproliferation to allo- or soluble antigens could be inhibited when T cells were cocultured with antigen presenting cells (APCs) pulsed with Trichosanthin (Tk), a plant protein purified from a Chinese medicinal herb Trichosanthes kirilowii Maximovich. In this paper, data are presented dealing with the mechanism by which the Tk functioned as a down modulator. APCs of human peripheral blood were first treated with one of inhibitors of antigen processing (chloroquine, leupeptin or cycloheximide), followed by pulsed with Tk, and then added into a T cell culture with stimulators PMA and A 23187. The suppression of lymphoproliferation was observed to be obviously diminished or totally disappeared. In contrast, when CyA was used to replace the Tk for pulse-treatment of APCs that had received the same pretreatment with the inhibitors, no significant change was detected for the suppression, suggesting that the Tk might use a different pathway to induce hyporeactivity and that the pathway was concerned in APC's function of antigen presentation. This view obtained support from our immuno-histochemical examination of the Tk-pulsed APCs. By preparing colloidal gold-labelled Tk (Tk-G particle), we were able to show that the Tk-G, when incubated with APCs and T cells at 37 degrees C for two hours and washed, was visualized under the electronic microscope to be bound to APCs', instead of T cells', membrane surface and internalized cells' into endosomes. And then the Tk-G particles were further identified within lysosomes. In this way, the Tk molecules were subjected to be processed as an external antigen and might be presented to T cells to activate certain T subsets that in turn mediated the immunosuppression. This course was capable of, as expected, being interrupted by antigen processing inhibitor, especially by chloroquine.

[Studies on the analgesic and antiinflammatory action of radix Linderae extract].

Both of aqueous extract and alconol extract of Radix Linderae at a dose 5 g/kg and 10 g/kg could length obviously pain threshold in mice in hot plate test. The samples at a dose 20 g/kg could inhibit significantly the writhing frequency induced by potassium antimony tartate in mice, also antagonize the swelling of ear induced by inflammatory agent and decrease the swelling rate. The component further isolated from the plant could antagonize the swelling of rat toes induced by carrageenin.

Hepatic angiomyolipoma: a report of two cases.

Two cases of rare hepatic angiomyolipoma are reported. Both represented with slight abdominal discomfort. The first patient complained of abdominal fullness, and had a normal physical examination and laboratory data. The second came with epigastralgia. Tenderness over right upper quadrant was noted and positive hepatitis B antigen was found. Ultrasound demonstrated hyperechoic hepatic lesion in both. Although diagnosis of hepatic angiomyolipoma was suspected by the radiological findings of ultrasound, CT, MRI, and angiography, it was confirmed by the histological presence of three mesenchymal components: abundant vessels, mature fat cells and smooth muscle cells. For fear of the coexistence of hepatocellular carcinoma, especially given the high prevalence area of Taiwan, surgical intervention was recommended if liver function permitted. Successful treatment was achieved by hepatic resection in both cases.

Spindle cell lipoma in a 14-month-old girl.

This case report concerns a girl with spindle cell lipoma of the neck. Spindle cell lipoma is a variant of lipomas and was first described by Enzinger and Harvey in 1975. It occurs chiefly in males between 40 and 70 years of age. It is a benign lesion that can be cured by excision, and local recurrence is rare. Spindle cell lipoma is composed of adipocytes and non-fat-storing immature mesenchymal cells. The condition is uncommon in adults and had not been reported to occur in children.

rRNA operon restriction derived taxa for Streptomyces (RiDiTS).

A method for grouping Streptomyces strains by fingerprints of their rRNA operons is described. In polyacrylamide gels, multicopy rRNA operon fragments in Streptomyces genomic MseI fingerprints produced intense bands which are well resolved from the less conspicuous low copy fragments interspersed between them. The high intensity multicopy rRNA bands are easily distinguished from the low intensity bands, eliminating the need for Southern blot hybridization to visualize the rRNA fragments. Direct evidence that the high-intensity bands in these polyacrylamide gels originated from rRNA operons was provided by a 'differential' Southern blot technique. We have used this method to assign 98 strains to 11 rRNA fingerprint type groups. This clustering method may be applicable to any prokaryote with a high G+C content genome.

[Sézary syndrome].

Sézary syndrome is a form of leukemia-lymphoma characterized clinically by erythroderma, pruritus, adenopathy, and circulating atypical cells with cerebriform nuclei. Histologically, atypical lymphocytes in the dermis and Pautrier's microabscesses are often present in skin biopsy specimens. We herein report a typical case of Sézary syndrome showing T-suppressor-cell characteristics, and related literatures are reviewed.