Animals ; Cells, Cultured ; Chondrocytes ; Interleukin-1beta ; NF-kappa B ; Osteoarthritis/drug therapy* ; Rats ; Tumor Necrosis Factor-alpha

Objective The aim of this study was to investigate the diagnostic impact of polymerase chain reaction (PCR) assays for fungal pathogens in fluid samples,and to evaluate the feasibility of fast PCR diagnostic method for the invasive fungal infection.Methods The sterility body fluid samples from 60 cases hospitalized immunocompromised patients with clinical underlying diseases and suspect of invasive infections with fungi between January 2008 and December 2011 were processed for microscopy and cultures.Applying fungal ITS4 and ITS86 universal primers to amplify pathogenic fungi genes from the sterility body fluid with method of rapid PCR.The results were compared between the standard and PCR methods.Results Humoral direct clinical specimens by PCR amplification of DNA fragments with the scan results were similar.Positive rate of PCR test with clinical body fluid samples and the traditional fungal cultivation was similar.There was no significant statistical difference [38.3％ (23/60) vs 33.3％ (20/60),P ＞ 0.05].Conclusions PCR test is feasibility with clinical fungal diagnosis from directly humoral specimens.To amplify the clinical sterility body fluid samples with ITS fungal universal primers and PCR method might provide an accurate and rapid approach to detect the pathogenic fungi.Its methods on early diagnosis and prognosis of invasive fungal infections are of guiding significance.

The inflammation factors and roles of them in acute coronary syndrome (ACS) were explored. The similarity between the theory of pathogenic toxin in Chinese Medicine and the inflammation response theory in ACS was discussed. The exploration of new inflammatory factors may be helpful for Chinese Medicine in the research of ACS.
Acute Coronary Syndrome ; complications ; diagnosis ; prevention & control ; therapy ; Humans ; Inflammation ; complications ; etiology ; pathology ; therapy ; Inflammation Mediators ; physiology ; Medicine, Chinese Traditional ; Prognosis ; Toxins, Biological ; adverse effects

Objective: To evaluate the effect of docosahexaenoic acid (DHA) on microglial activation during oxygen-glucose deprivation and restoration (OGD/R) injury. Methods: N9 microglia were inoculated in 96-well culture plates at a density of 10⁴ cells/well for 3-5 days and divided into 3 groups (n=18 each) using a random number table method: control group (group C), group OGD/R and DHA+ OGD/R group. The cells were cultured for 24 h in an incubator at 37 ℃ (95% air-5% CO₂) in group C. In OGD/R and DHA groups, the culture medium was replaced by glucose-free culture medium, the cells were cultured for 12 h in an incubator at 37 ℃ (95% N₂-5% CO₂), and then cells were returned to the normal culture medium and cultured for 24 h in an incubator at 37 ℃ (95% air-5% CO₂) to establish the OGD/R injury model. The cells in group DHA were incubated with 25 μmol/L DHA at 12 h before OGD/R. The cell viability was detected using the methyl thiazolyl tetrazolium assay, the activity of lactate dehydrogenase (LDH) was measured by using chemical colorimetric method, activated microglia were counted by immunofluorescence staining, the expression of microglia activation marker iba-1 was detected by Western blot, and the concentrations of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and IL-4 and IL-10 in culture medium were determined using enzyme-linked immunosorbent assay. Results: Compared with the group C, the cell viability was significantly decreased, the LDH activity was increased, the number of activated microglia was increased, and the expression of iba-1 was up-regulated in OGD/R and DHA+ OGD/R groups, the concentrations of TNF-α and IL-6 were significantly increased in group OGD/R, and the concentrations of TNF-α, IL-6, IL-4 and IL-10 were significantly increased in group DHA+ OGD/R (P<0.05). Compared with group OGD/R, the cell viability was significantly increased, the LDH activity was decreased, the number of activated microglia was decreased, the expression of iba-1 was down-regulated, TNF-α and IL-6 concentrations were decreased, and IL-4 and IL-10 concentrations were increased in group DHA (P<0.05). Conclusion The mechanism by which DHA reduces OGD/R injury may be related to inhibiting activation of microglia and to reducing inflammatory responses.

Objective To explore the relationship between ERG4 gene overexpression and azole resistance in clinical Candida albicans strains. Methods The National Committee for Clinical Laboratory Standards (NCCLS)M27-A2 broth microdilution method was conducted to evaluate antifungal susceptibility of 34 clinical Candida albicans isolates in vitro. Total RNA was extracted from these Candida albicans strains and transcribed into cDNA. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of ERG4 gene. Statistical analysis was carried out by a two-sample t-test. Results The expression level of ERG4 mRNA was significantly higher in fluconazole-resistant than in -sensitive Candida albicans strains (4.20 ± 2.56 vs. 1.72 ± 1.33, t = 3.99, P < 0.05), higher in itraconazole-resistant than in -sensitive Candida albicans strains (3.60 ± 2.47 vs. 1.66 ± 1.61, t = 3.71, P < 0.05), and higher in voriconazole-resistant than in -sensitive Candida albicans strains (3.99 ± 2.72 vs. 2.07 ± 1.58, t = 2.91, P <0.05). Further more, increased ERG4 mRNA expression was also observed in isolates cross-resistant to all the three azole antifungal agents compared with those susceptible to all of them (4.49 ± 2.73 vs. 1.69 ± 1.82, t = 3.81, P < 0.05). Conclusions The overexpression of ERG4 gene may be associated with cross resistance to fluconazole, itraconazole and voriconazole in clinical Candida albicans strains, but its exact role is expected to be investigated through downregulation of the ERG4 gene.

Objective To determine the role of interleukin-8 (IL-8) produced by tumor induced fibroblasts in the development of cutaneous melanoma. Methods B16 melanoma cells induced L929 fibroblasts phenotype was transdifferentiated to myofibroblasts (MF) by co-culture in vitro. MF was monitored by morphology and immunophenotype for a-SMA. The level of IL-8 was detected by ELISA. The effect on B16 cell proliferation rate was estimated using MIT method in vitro. Melanoma implanting model was constructed in C57 mice. Results L929 MF phenotype could be modulated by B16 melanoma cells-derived transforming growth factor-β1 (TGF-β1) and elevated the levels of IL-8. L929 MF did not influence the B16 melanoma cells viability in vitro, but shortened the time of tumor formation and increased the incidence rates of tumors in C57 implanting model mice. Conclusion Fibroblasts can be activated by tumor cells and produce IL-8, which acts as an inflammatory cytokine promoting the development of cutaneous melanoma.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastritis, Atrophic ; diagnosis ; pathology ; Gastroscopy ; methods ; standards ; Humans ; Male ; Middle Aged ; Specimen Handling ; methods ; Tissue Embedding ; methods

Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Child ; Female ; Gene Expression Regulation, Neoplastic ; Hodgkin Disease ; genetics ; metabolism ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Staining and Labeling ; methods ; Young Adult

In order to summarize and perfect the basic skill practicing method in acupuncture teaching and form a characteristic teaching, the research on skill training and technique improving of acupuncture and tuina major was carried out. The undergraduate students in acupuncture and tuina major from College of Acupuncture-Moxibustion and Tuina were randomly divided into an experimental group and a control group. Students from both groups received the basic courses of acupuncture-moxibustion and tuina as requested in the syllabus and the special teaching was added to the experimental group, namely "three practice" (strength practice, coordination practice, targeted practice of acupuncture basic skills) and "three enlightenment" (technique enlightenment, mind and qi enlightenment, efficacy enlightenment). During and after the courses, the teaching effectiveness was evaluated on the basis of needling manipulation. The peacetime score in the experimental group was superior to that in the control group (38.03 +/- 1.14 vs 33.25 +/- 1.31, P < 0.05), the total score in the experimental group was superior to that in the control group (84.03 +/- 6.30 vs 78.05 +/- 6.55, P < 0.05). The special teaching method of "three practice" and "three enlightenment" can improve the effect of skill training teaching method. It highlights the unique feature of central plains by integrating the essence of Shaolin internal qigong and Chen-style Taiji, it can also provide new ideas for the practical teaching of TCM professional courses and the construction of characteristic professionals.
Acupuncture ; education ; manpower ; Acupuncture Therapy ; methods ; Humans ; Students ; Teaching

The present work is to investigate the chemical constitutions of Hyptis rhomboidea and their antifungal activities. The compounds were isolated by Toyopearl HW-40, Sephadex LH-20, MCI-Gel CHP-20, RP-18, PTLC and silica column chromatographic methods and subjected to evaluate some monomers antifungal activity of eight kinds of plant pathogenic bacteria. Eleven compounds were isolated and identified as ethyl caffeate (1), ursolic acid (2), oleanolic acid (3), vanillactic acid (4), methyl rosmarinate (5), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1 --> 6) -beta-D-glucopyranoside (6), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (7), ilexgenin A (8), beta-amyrin (9), kaempferol 3-O-beta-D-glucopyranoside (astrgalin, 10) and cholest-5-ene-3beta, 4beta-diol (11). Compound 1 showed the strongest inhibitory effect on Sclerotinia sclerotiorum with the MIC 16.2 mg x L(-1), and compound 5 showed the strongest inhibitory effect on S. minor and Exserohilum turcicum with MIC 16.2, 8.1 mg x L(-1), respectively. All compounds were isolated from the H. rhomboidea for the first time, and compounds 1 and 5 showed antifungal activity.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fungi ; drug effects ; Hyptis ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization