In the present study, terminal-restriction fragment length polymorphism (T-RFLP) technique was applied to assess the diversity and tissue distribution of the fungal endophyte communities of Alpinia officinarum collected from Longtang town in Xuwen county, Guangdong province, China, at which the pharmacological effect of the medicine plant is traditional considered to be the significantly higher than that in any other growth areas in China. A total of 28 distinct Terminal-Restriction Fragment (T-RFs) were detected with HhaI Mono-digestion targeted amplified fungal nuclear ribosomal internal transcribed spacer region sequences (rDNA ITS) from the root, rhizome, stem, and leaf internal tissues of A. officinarum plant, indicating that at least 28 distinct fungal species were able to colonize the internal tissue of the host plant. The rDNA ITS-T-RFLP profiles obtained from different tissues of the host plant were obvious distinct. And the numbers of total T-RFs, and the dominant T-RFs detected from various tissues were significantly different. Based on the obtained T-RFLP profiles, Shannon's diversity index and the Shannon's evenness index were calculated, which were significantly different among tissues (P < 0.05). Furthermore, two types of active chemicals, total volatile oils by water vapor distillation method and galangin by methanol extraction-HPLC method, were examined in the each tissue of the tested plant. Both of tested components were detected in all of the four tissues of the medicine plant with varying contents. And the highest was in rhizome tissue. Correlation analysis revealed there were significant negative correlations between both of the tested active components contents and calculated Shannon's diversity index, as well as the Shannon's evenness index of the fungal endophyte communities of the host plant (P = 0, Pearson correlation coefficient ≤ -0.962), and significant positive correlations between both of the tested active components contents and 325 bp dominant T-RF linkage to Pestalotiopsis (P = 0, Pearson correlation coefficient ≥ 0.975). In conclusion, A. officinarum is colonized by diverse fungal endophytes communities. The diversity of the fungal endophytes was found in the A. officinarum varied with differences of the tissue types of the host plants and was closely correlated with the accumulation of main active components, total volatile oils and galangin contents in the host plant tissue.
Alpinia ; chemistry ; microbiology ; Biodiversity ; China ; DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Drugs, Chinese Herbal ; analysis ; Endophytes ; classification ; genetics ; growth & development ; isolation & purification ; Fungi ; classification ; genetics ; growth & development ; isolation & purification ; Phylogeny ; Plants, Medicinal ; chemistry ; microbiology ; Polymorphism, Restriction Fragment Length

Objective To observe the levels of serum leptin in Gaves disease(GD)and thyroiditis(HT)Datients and to discuss the immunological role of leptin in the pathogenesis of autoimmune thyroid disease(AITD).Methods 102 newly diagnosed female AITD patients were divided into 3 groups:GD hyperthyroid group,HT hypothyroid group and subclinical hypothyroid group.Age,sex and BMI-matched 27 euthyroid,healthy subjects served as controis.The levels of FT3,FT4 and sTSH were determined by immunofluorometrie assay.ELISA kit was aDplied to measure the levels of serum leptin.Results Serum FT3 and FT4[(19.74±15.39),(78.25±58.68)pmol/L]levels of GD hyperthyroid patients were obviously higher than those of the controls[(4.87±0.25),(15.96±3.15)pmol/L,P＜0.01],but serum sTSH and leptin levels[(0.15±0.08)mU/L,(8.73±1.92)μg/L]were obviously lower than those of the controls[(3.81±0.19)mU/L,(12.38±3.51)μg/L,P＜0.01or＜0.05].Serum FT3 and FT4[(3.36±0.26),(6.95±3.29)pmol/L]levels of HT hypothyroid patients were obviously lower than those of the controls(P＜0.05),but serum sTSH and leptin levels[(45.48±35.83)mU/L,(17.17±3.82)μg/L]were obviously higher than those of the controls(P＜0.01 or＜0.05).Serum FT3 and FT4[(4.67±0.60),(14.87±2.14)pmol/L]levels of subclinical hypothyroid patients had not statistical difference comparing with those of the controls(P＞0.05),but serum sTSH and leptin levels[(13.67±8.66)mU/L,(16.25±3.67)μg/L]were obviously higher than those of the controls(P＜0.01 or＜0.05).Conclusions Leptin might have an immuoregulation role in the pathogenesis of AITD.In addition,serum levels of leptin in AITD is also influenced by many other related hormones.

Antimicrobial peptides, a cluster of small peptides secreted by the majority of creatures, have been demonstrated with activity against a wide range of microorganisms including bacteria, protozoa, yeast, fungi, viruses and even tumor cells. These peptides have some features such as broad spectrum , high effi-cacy and stability, little drug resistance. A lack of new antibiotics combined with emerging multi-drug resis-tance issues demands that new antimicrobial strategies be explored for treating these infections. It has been proposed that the antimicrobial peptides might form the foundation for a new class of clinically useful an-timicrobials. We review the advantages of these molecules in construction features and bioactivity, with the focus on the mechanism and clinical applications.

Objective To explore the effect of iodine on the pathogenesis of postpartum thymiditis.Methods Forty-four female C57BL/6J mice,8-week old,fed by low iodine dietary(the concentration of iodine≤35 μg/kg),were randomly divided into 4 groups:non-pregnancy experimental autoimmune thymiditis(non-pregnancy EAT)group with 8 mice,EAT of mice was induced by immunization with pig's thyroglobulin(Tg)in the presence of complete Freund's adjuvant.Six mice in non-pregnancy EAT group survived at the end of experiment;normal iodine-PPT(NI-PPT)group,10-fold high iodine-PPT(10HI-PPT)group and 50-fold high iodine-PPT(50HI-PPT)group with 12 mice in each group.The last 3 groups mice,who received the same immunization schedule as the above,were mated with adult male mice followed by induction of EAT.In the end,7,6 and 6 mice were noticed to be pregnant in each group.All animals were killed 4 weeks after postpartum.Histological severity of thyroid specimens was evaluated.The serum level of thyroglobulin antibody(Tg-Ab),thyroid pomxidase antibody(TPO-Ab),TT3 and TT4 were measured by radioimmunoassay(RIA).The expression level of IFN-γ/IL-4 mRNA in spleenwere assayed by RT-PCR.Results Pathological examination showed the infiltration of inflammatory cells.epithelial cell applanation,follicle atrophy or destruction.The severity of inflammation in non-pregnancy EAT bgroup.NI-PPT group and 10HI-PPT group was less serious than that in the 50HI-PPT group,the difference has bstatistical significance(P＜0.05).The level of TPO-Ab in non-pregnancy EAT group,NI-PPI-group,10HI-PPTgroup and 50HI-PPT group wag(14.32±8.85)%,(64.45±10.52)%,(38.46±5.57)%and(90.09±9.98)%.respectively the difference being statistically significant between any two groups(P＜0.05).There was no statisticaldifference(F＝0.484,P＞0.05)of Tg-Ab among non-pregnancy EAT group[(33.74±3.71)%],NI-PPT group [(29.65±2.06)%],10HI-PPT group[(37.21±3.87)%]and 50HI-PPT group[(33.87±4.17)%].There was no statistical difference(F＝1.596,P＞0.05)of TT3 among non-pregnancy EAT group (2.47±0.69)%,NI-PPT group(1.57±0.25)%,10HI-PPT group[(1.60±0.28)%]and 50HI-PPT group[(1.82±0.75)%].The level of TT4 in 50HI-PPT group[(66.68±5.47)%]was lower than that in non-pregnancy group,NI-PPT group and 10HI-PPTgroup[(99.87±5.97)%,(89.13±7.64)%and(91.05±5.82)%],the difference being statistically significant(P＜0.05).The expression level of IFN-γ mRNA was increasing,being 1.02±0.10,1.37±0.10,1.39±0.12 and 1.68±0.06 in non-pregnancy EAT group,NI-PPT group,10HI-PPT group and 50HI-PPT group.The difference had a statistical significance between any two groups except for NI-PPT group and 10HI-PPT group(P＜0.05).The expression level of IL-4 mRNA in 10HI-PPT group(0.49±0.04)and 50HI-PPT group(0.53±0.06)were all higher than non-pregnancy EAT group(0.24±0.05)and NI-PPT group(0.35±O.05),the differences being statistically significant (P＜0.05).Conclusions Adequate iodine supplementation during pregnancy and postpartum period is necessary,but iodine excess could induce postpartum thyroiditis.So iodine supplementation during pregnancy and postpartum should be adequate and reasonable.

AIMTo study the effect of taurine (Tau) on rabbit cardiomyocyte apoptosis during ischemia/reperfusion (I/R) injury.

METHODSRabbit heart I/R injury was induced by occluding the left anterior descending coronary artery for 45 min and reperfusion for 180 min. taurine (200 mg/kg) was intravenously injected 5 min before heart ischemia. Cardiomyocyte apoptosis was measured by using terminal deoxynucleotidyl transferase--mediated dUTP nick end labeling method (TUNEL), flow cytometry (FCM) and DNA agarose gel electrophoresis.

RESULTSDNA ladder pattern of DNA in myocardium was revealed by agarose gel electrophoresis in I/R group while was not found in Tau + I/R group. Apoptotic cardiomyocytes were sparse within ischemic myocardium at risk in Tau + I/ R group as compared with that in I/R group (TUNEL stain). Apoptosis rate in ischemic myocardium from I/R and Tau + I/R groups detected by flow cytometry was 17.66% +/- 1.54% and 4.86% +/- 1.23%, respectively. Fas and Bax protein expressions in ischemic myocardium of I/R group were higher than that in nonischemic myocardium group (P < 0.01), Bcl-2/Bax ratio in I/R group was lower than that in nonischemic myocardium (P < 0.01); while in Tau + I/R group, Fas and Bax protein expressions were lower than that in I/R group (P < 0.01), the Bcl-2/Bax ratio was higher than that in I/R group (P < 0.01).

CONCLUSIONTaurine reduced apoptosis of myocytes in I/R rabbit heart; its mechanism may involve Fas, Bax and Bcl-2 proteins expression.

Animals ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Male ; Myocardial Ischemia ; metabolism ; pathology ; Myocytes, Cardiac ; cytology ; drug effects ; Rabbits ; Reperfusion Injury ; metabolism ; pathology ; Taurine ; pharmacology

Objective To explore the changes in neuroglobin(NGB)expression in rat cerebral cortex induced by acute and chronic hypoxia at high altitude.Methods Seventy SD rats were randomly divided into normal control and experimental groups,and in the latter group,the rats kept in a high-altitude research base in Kekexili(4600 m),while the control rats were kept in a facility at the altitude of 2295 m.The rats in the experimental group were divided into 6 groups with the exposure time of 12,24,48,72 h,1 week and 1 month.An oximeter was used to measure the SaO2 level.Semi-quantitative PCR and Western blotting were performed to detect the expression levels of NGB mRNA and protein in the cortical neurons of the rats after the exposure.Results After explosure of the rats to hypoxia at high altitude for 12h,the SaO2 was lowered to(70.70±2.83)%and increased gradually as exposure time prolonged,but remained lower than that in the control group throughout the exposure.RT-PCR showed a rapid increase of NGB mRNA expression after 24-h exposure to hypoxia,followed by gradual decrease till recovery of the normal level at 1 week;the expression slowly increased after 1 week and maintained a high level till 1 months.showing significant difference from that in the control group(P＜0.05).Western blotting showed an identical pattem of NGG protein expression alterations during the experiment.Conclusion NGB expressions in the cerebral cortex increase significantly after acute and chronic hypoxia at an altitude of 4600 m to enhance the tolerance to hypobaric hypoxia,suggesting the possible role of NGB as an important endogenous mechanism for protecting the neural tissues against hypoxic injuries.

OBJECTIVETo investigate the significance of clinicopathological stage of chronic idiopathic myelofibrosis (CIMF) in WHO classification of 2001.

METHODSHistopathological analysis of bone marrow biopsy plastic-embedded sections stained with H-G-E and Gomori's stains and clinical features of 113 cases previously diagnosed as primary myelofibrosis (PMF) and 48 cases MPD-U (total of 161 cases which including male 79 and female 82) were studied retrospectively.

RESULTSThere was no significant differences on the clinical features among the cellular phase, collagen fiber phase, sclerotic phase and osteomyelosclerosis of 113 previously diagnosed patients. According to WHO classification 2001 of CIMF, previously diagnosis in 48 cases with MPD-U was WHO pre-CIMF, and in 113 cases with PMF was WHO CIMF-Fs. There were significant differences between of WHO pre-CIMF and WHO CIMF-Fs about clinicopathological features except age. The percentage of immature granulocytes, normoblasts, lymphocytes in peripheral blood, the size of hepatosplenomegaly, and the percent age of tear drop-like red blood cells in pre-CIMF were significantly lower than those in CIMF-Fs (P < 0.05). However, the number of hemoglobin and platelets in patients with pre-CIMF were significantly higher than that with CIMF-Fs (P < 0.01).

CONCLUSIONpre-CIMF and CIMF-Fs in clinical and histopathological features were different development stage of CIMF, while osteomyelosclerosis is a variant of CIMF, but not an independent disease.

Adult ; Aged ; Biopsy ; Bone Marrow ; pathology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Primary Myelofibrosis ; classification ; pathology ; Thrombopoiesis

OBJECTIVETo assess protein and mRNA expression levels of heparanase and basic fibroblast growth factor (bFGF) genes in human non-small cell lung cancer (NSCLC) and their roles in tumor invasion, metastasis and prognosis.

METHODSA total of 115 paraffin-embedded and 45 fresh-frozen tissue specimens of NSCLC were studied by immunohistochemistry, Western Blot and in situ hybridization to evaluate the protein and mRNA expression status of heparanase and bFGF genes. The data was analyzed by SPSS statistical software.

RESULTSBoth human heparanase and bFGF were highly expressed in NSCLC cells, in contrast to none or a low expression in normal lung tissue. Expression of heparanase also showed a significantly higher than that in the normal tissue by Western blot (P = 0.041). Immunohistochemistry showed that heparanase expression was both cytoplasmic and membranous. The agreement between heparanase and bFGF was significant. A significant correlation was found between the expression of either protein and TNM stage, vascular invasion, lymphatic metastasis and microvascular density (MVD). Co-expression of the two proteins demonstrated an even higher correlation with the tumor stage and MVD. In addition, expression of bFGF correlated with tumor cell differentiation. Data of a multivariate analysis indicated that tumor cell differentiation, vascular invasion, lymphatic metastasis and expression of bFGF were identified as significant prognostic parameters.

CONCLUSIONSBoth heparanase and bFGF may play important roles in tumor angiogenesis, metastasis, and prognosis of NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; enzymology ; metabolism ; pathology ; Cell Differentiation ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Glucuronidase ; metabolism ; Humans ; Lung Neoplasms ; enzymology ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; Prognosis ; Survival Rate

OBJECTIVETo explore treatments of severe post-kidney-transplant lung infection by integrative Chinese and Western medicine (ICWM), in order to elevate the curing rate as well as to lower the death rate.

METHODSBased on conventional ways of Western medical treatments of 18 cases of severe post-kidney-transplant lung infection, such as putting the patients in single individual ward, antibiotics to prevent infection, respiratory machines, blood filtration, nutritional support, steroids, and maintaining electrolytes balance, we applied integrated Chinese medicinal treatments, like altering conventional prescription "pneumonia III", and conducted clinical observation of effectiveness, and indexes including white blood cell (WBC), neutrophilic granulocyte, blood urea nitrogen (BUN), blood creatinine (Cr), etc.

RESULTSOf the 18 cases studied, 7 were already cured, 8 proved the treatment effective, 3 died. All clinical indexes had statistically significant changes compared with those of before treatment (P < 0.01).

CONCLUSIONICWM can increase curing rate and lower death rate.

Adult ; Aged ; Anti-Bacterial Agents ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunocompromised Host ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; Lung Diseases ; drug therapy ; immunology ; Male ; Middle Aged ; Opportunistic Infections ; drug therapy ; Respiratory Tract Infections ; drug therapy ; immunology

OBJECTIVETo explore the response of rat bone marrow mesenchymal stem cells (MSCs and calvarial osteoblasts to mechanical strain and the consequent changes of cytoskeleton F-actin.

METHODSBone marrow MSCs and calvarial osteoblasts were isolated from SD rats and cultured in vitro. Mechanical stretch was performed on passage 3 cells at 2 000 microepsilon for 0, 2, 6 and 12 hours using four-point bending system. The response of cells and the distribution of F-actin were observed using fluorescent staining under laser scanning confocal microscope and the morphological parameters were quantified using image analysis software Laserpix.

RESULTSUnder mechanical stretch, the fluorescent staining decreased obviously at both MSCs and osteoblasts, and F-actin filaments were rearranged and became tenuous, thinner, and abnormally distributed. The outline of nucleus became unclear and apoptotic changes were observed at some of both cells. Cellular size decreased more significantly in MSCs than in osteoblasts. Quantity analysis showed that total area of cells, total fluorescent density and green fluorescent density (F-actin) were all significantly decreased in MSCs (P < 0.05 or P < 0.01), and total fluorescent density, green fluorescent density and red fluorescent density (nuclei) did also in osteoblasts (P < 0.05 or P < 0.01).

CONCLUSIONMechanical stretch caused extensive response on both MSCs and osteoblasts which led to the rearrangement of F-actin filament and apoptosis in some of these cells. MSCs were more sensitive to mechanical strain than osteoblasts.

Actin Cytoskeleton ; metabolism ; Actins ; metabolism ; Animals ; Bone Marrow Cells ; Cells, Cultured ; Cytoskeleton ; Mesenchymal Stromal Cells ; Microtubules ; Osteoblasts ; Rats ; Stress, Mechanical