Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Purpura, Thrombotic Thrombocytopenic ; etiology ; Transplantation, Autologous

OBJECTIVETo reveal the quality variation of polysaccharides, triterpenoids and proteins in spores and fruiting bodies of Ganoderma lucidum from producing areas, different varieties, harvesting parts and periods, and wall-breaking treatments.

METHODSpores and fruiting bodies from varieties of Longzhi No. 1 and Hunong No. 1 were collected as test samples, together with wall-broken spores sold in domestic main producing areas. The anthrone-sulfuric acid colorimetric method was used to determine the content of total polysaccharides. The vanillin-glacial acetic acid-perchloric acid colorimetric method was used to determine the content of total triterpenoids. The Lowry method was used to determine the content of total proteins.

RESULTThe content ranges of total polysaccharides, total triterpenoids, and total proteins from 6 domestic main producing areas were 0.40% - 2.25%, 1.36%-3.15% and 0.74% -1.91% respectively. The content ranges of total polysaccharides, triterpenoids, and proteins in the fruiting bodies from 2 varieties cultured in Zhejiang were 0.25% -1.42%, 0.44% -1.42% and 1.82% -3.67% respectively. In addition, the ranges of samples from wall-unbroken spores were 0.41% - 0.91%, 0.09% - 0.12%, 0.78% - 0.90% respectively and wall-broken spores are 1.03% - 2.25%, 1.89% - 3.15%, 0.96% - 1.04% respectively.

CONCLUSIONThere are significant differences in the contents of main chemical ingredients of wall-broken G. lucidum spores saled in the markets. The samples from Zhejiang contain high content of total polysaccharides and triterpenoids, and samples from Fujian contains more proteins. Between the 2 major varieties cultured in Zhejiang, Longzhi No. 1 contains higher content of triterpenoids, but Hunong No. 1 has more polysaccharides. Contents of triterpenoids and polysaccharides from wall-broken spores are much higher than those of fruiting bodies. The stipes from fruiting bodies contains more polysaccharides than those of the pileus, while the triterpenoids contents are higher in the pileus than stipes. The pileus and stipes collected in the second year contain higher content of polysaccharides than the first year's samples, but the contents of triterpenoids are lower. Wall-breaking treatment would significantly improve the extraction and dissolution rate of total triterpenoids and polysaccharides.

Fungal Proteins ; analysis ; Polysaccharides ; analysis ; Reishi ; chemistry ; Spores, Fungal ; chemistry ; Triterpenes ; analysis

AIM:To observe the situation of rat retinal tissue DNA damage at early diabetic period, discuss the role of the blood glucose fluctuations, and provide a new method for studying the pathogenesis of diabetic retinopathy ( DR) .
METHODS: SD rats were randomly divided into four groups:normal control group (NC), normal fluctuation group ( NF ) , diabetes group ( DM ) and diabetes fluctuation group ( DF ) . Diabetic models were established through intraperitoneal injection of STZ. A certain amount of glucose was injected in the rats of group NF and DF in an intraperitoneal mode three times a day after the model was established, thereby causing blood glucose fluctuations. Rats were killed and the retinal tissues were taken in the 8th week. Single cell gel electrophoresis ( SCGE ) technique was adopted for detecting DNA injury extent in the retina tissue.
RESULTS:Groups NF and DF showed significant and regular fluctuations. The curve of blood glucose fluctuations was relatively stable. All values of MBG, SDBG, LAGE and M were significantly increased compared with group NC. Group DF was increased more significantly. It was statistically significant (P<0. 01). SCGE showed that there were DNA damages in different levels in the cells of group NF, DM and DF. Indicators of cells such as TL, TDNA %, TM, OTM were higher than that in group NC. It was statistically significant ( P<0-01). The comparison difference between two groups was also significant (P<0. 01).
CONCLUSION: Rat retinal tissues have DNA injury during early diabetic period. DNA injury is gradually aggravated with blood glucose fluctuation. It indicates that high blood glucose and blood glucose fluctuation are involved in the mechanism of cell DNA injury, and they may be one of DR early event, have played a certain role in the incidence of DR.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

OBJECTIVETo compare the effect of citric acid stimulation on salivary alpha-amylase (sAA), total protein (TP), salivary flow rate, and pH value between Pi deficiency (PD) children and healthy children, thereby providing evidence for Pi controlling saliva theory.

METHODSTwenty PD children were recruited, and 29 healthy children were also recruited at the same time. Saliva samples from all subjects were collected before and after citric acid stimulation. The sAA activity and amount, TP contents, salivary flow rate, and pH value were determined and compared.

RESULTS(1) Citric acid stimulation was able to significantly increase salivary flow rate, pH value, sAA activities, sAA specific activity and sAA amount (including glycosylated and non-glycosylated sAA amount) in healthy children (P<0.05), while it could markedly increase salivary flow rate, pH value, and glycosylated sAA levels in PD children (P<0.05); (2) Although there was no statistical difference in determined salivary indices between the two groups (P>0.05), salivary indices except salivary flow rate and glycosylated sAA levels decreased more in PD children. There was statistical difference in sAA activity ratio, sAA specific activity ratio, and the ratio of glycosylated sAA levels between PD children and healthy children (P<0.05).

CONCLUSIONPD children had decreased response to citric acid stimulation.

Child ; Citric Acid ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Saliva ; Salivary alpha-Amylases ; metabolism ; alpha-Amylases

Objective To investigate the effect of interferon alpha ( IFN-α) on apoptosis of vascular smooth muscle cells ( VSMCs) in rats and the related mechanism. Methods The cells were divided into three group:group A, group B and group C.Group A was transfected with nonspecific siRNA, group B was intervened with IFN-α and transfected with nonspecific siRNA, and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).P204, RAS protein levels, and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC/PI method. Results As compared with group A, the expression of P204 mRNA and protein in group B was up-regulated (P<0.05), the cell apoptosis was increased (P<0.05), in the process of the above, the expression of RAS protein was decreased ( P<0.05) and the phosphorylation levels of RAF and ERK were dropped (P<0.05).In group C, the expression levels of P204 mRNA and protein were down-regulated (P<0.05), and cell apoptosis was decreased ( P<0.05) , the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased ( P<0.05) . Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.

AIM: To evaluate the effects of the extract of Buddleja officinalis eye drops in basic tears secretory volume, tear film stability, expression of androgen receptors(AR) in castrated rats with dry eye, and to investigate the therapeutic effects of extract of Buddleja officinalis on dry eye caused by gonadal hormones level imbalance. METHODS:A total of 45 Wistar masculinity rats were divided at random into 9 groups, including normal group(A1,A2 and A3), model group(B1,B2 and B3), therapy group with extract of Buddleja officinalis eye drops(C1,C2 and C3). The "1" stood for being fed for 1 month, and "2" for 2 months, and "3" for 3 months. The dry eye model was established with orchiectomy on group B,C. Group C was treated with Buddleja officinalis extract eye drops for one month. All rats were checked with Schirmer Ⅰ test (SⅠt) and tear film break-up time (BUT). Expression of AR was analyzed by flow cytometer(FCM). RESULTS:The SⅠt value of group C was significantly higher than that of group B (P<0.01) and the BUT value of group C was significantly longer than that of group B (P<0.01), which indicated the eye drop could significantly keep basic tears secretory volume and tear film stability. And the expression of AR of group C was much higher than that of group B,which showed that available composition of the eye drops maybe display androgen-like activity.CONCLUSION:The main components of extract of Buddleja officinalis is the flavonoids which could significantly inhibit happening of dry eye of rat after androgen level lowered. Its mechanism is like androgen's and it could display androgen-like activity to keep basic tears secretory volume and tear film stability.

Arachidonic acid is an essential fatty acid in human nutrition and a biogenetic precursor of the biologically active prostaglandins and leukotrienes. ?~5 desaturase catalyzes the ?~5 dehydrogenation of di-homo-?-linolenic acid to form arachidonic acid in biosynthetic pathway of arachidonic acid. Using real time PCR technology,the transcriptional expression levels of gene encoding ?5 desaturase in three Mortierella alpina strains M10,M6 and M23,and in different growth phase of high arachidonic acid yielding strain M6,were determined. Results showed that there was a distinct corelationship between mRNA transcript level of ?~5 desaturase gene and biosynthesis of arachidonic acid. Results indicated that ?~5 desaturase plays an important role in arachidonic acid biosynthesis.

To compare the microcalorimetric fingerprint profiles of Lonicerae japonicae Flos (Lj.F) and Lonicerae Flos (L.F), microcalormietry was applied to find the heat change regularity of Bacillus shigae (B. shigae) metabolism affected by Lj.F and L.F (we choose Lonicera macranthoides Hand.-Mazz in this paper) with different concentrations. The thermogenic curves and thermodynamics parameters were investigated as evaluation index, and then the date of experiment was studied by similarity analysis. All the results indicated that the Lj.F and Lonicera macranthoides Hand.-Mazz (L.m.H-M) significantly impacted the microbial growth and had good similarity in its inhibitory activities. The combination approach of chemical analysis with bioassay was developed and employed to ensure the safety and efficacy of Chinese herbal medicines.
Anti-Bacterial Agents ; isolation & purification ; pharmacology ; Bacillus ; drug effects ; growth & development ; Calorimetry ; methods ; Chemical Safety ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Flowers ; chemistry ; Lonicera ; chemistry ; classification ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Thermodynamics ; Thermogenesis

OBJECTIVETo explore the effect of vinyl chloride on reproductive and endocrine system of male rats.

METHODSMale SD rats were administered with vinyl chloride at dose of (0, 10, 100, 1000 mg/kg) for 14 and 28 days, respectively. The levels of testosterone (T), inhibin B, luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol (E2) were measured in serum and testis homogenates. Histopathological examinations were performed for testis with electron microscopy.

RESULTSCompared with the control group after 14-day exposure, T and E2 serum levels of 1000, 100, 10 mg/kg groups decreased, InhB and LH levels of three dose groups increased. LH serum levels of 100 mg/kg increased significantly statistically compared with control group (P < 0.05). After 28-day exposure, T serum levels of 100, 1000 mg/kg groups were (10.90 +/- 1.56), (8.52 +/- 2.85) ng/ml respectively (P < 0.05), InhB serum levels of 100, 1000 mg/kg groups were (31.40 +/- 6.21), (28.39 +/- 5.67) pg/ml respectively. Both of T and InhB serum levels of 100, 1000 mg/kg groups decreased significantly (P < 0.05). Serum FSH levels of 10, 100, 1000 mg/kg groups decreased significantly compared with control group (P < 0.05). Compared with groups of 14-day exposure, serum InhB and LH levels of 10, 100, 1000 mg/kg groups decreased significantly statistically after 28 days. T and InhB testis levels of 100, 1000 mg/kg groups were 8.05 +/- 2.19),(6.75 +/- 1.94) ng/mg pro and (39.32 +/- 5.55), (35.53 +/- 8.71) pg/mg pro respectively, which decreased significantly compared with control group (P < 0.05). Leydig cell and Sertoli cell were damaged according to histopathological examinations.

CONCLUSIONVinyl chloride has adverse effects on reproductive and endocrine system of male rats and may change their serum and testis homogenate levels of hormones.

Animals ; Follicle Stimulating Hormone ; blood ; Leydig Cells ; ultrastructure ; Male ; Rats ; Sertoli Cells ; ultrastructure ; Testis ; drug effects ; metabolism ; Testosterone ; blood ; Vinyl Chloride ; toxicity