Streptomyces sp. A048 was cultured in a complete medium to the last stage of log phase,the hyphae were washed and collected by centrifugation. Then the hyphae were inoculated in liquid medium for chitinase production using two-step fermentation. Activity of chitinase produced by two-step fermentation was 1.1 times higher than that from one-step fermentation,and ferment cycle was for 54 hours,which was 66 hours shorter than that of one-step fermentation. The hyphae and the powder of chitin were co-immobilizated and cultured in liquid medium for 36 hours,activity of chitinase was 1.8 times higher than that from one-step fermentation,and ferment cycle was 54h shorter than that of one-step fermentation. By adding 0.4% cellulose to two-step fermentation,activity of chitinase was 18.52 U/mL that was 4 times higher than that from the control and 10 times higher than that from one-step fermentation. Two step fermentation with chitin and cellulose may be the optimal fermentation process to produce Chitinase from Streptomyces sp. A048.

Syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid are three main bioactive ingredients in herbs of Saussurea involucrata with various pharmacological properties, while their contents are very low. In this study, the biosynthesis of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid in the cell suspension cultures of S. involucrata were regulated by feeding carbon sources and precursors, which resulted in a great increase of the contents and yields of the above three bioactive ingredients. After 16 days of fermentation, the yields of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid reached 339.0, 225.3, 512.7 mg x L(-1), respectively. Meanwhile, their contents increased up to 67.9, 1.9, 10.6 times of wild medicinal material, respectively. The results provided a solid basis for further studies on application of cell suspension cultures of S. involucrata for large-scale production of bioactive compounds syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid.
Cell Culture Techniques ; Cells, Cultured ; Chlorogenic Acid ; analysis ; metabolism ; Cinnamates ; analysis ; metabolism ; Glucosides ; analysis ; biosynthesis ; Phenylpropionates ; analysis ; Saussurea ; chemistry ; growth & development ; metabolism

From original concept and literature of acupoint, the concept and clinical significance of ashi method is discussed, which clarifies that the essence of ashi method is to locate the acupoints by patients' sensitivity on force. The clinical application of heat-sensitive moxibustion has illustrated that positioning method of this therapy is based on the appearance of heat-sensitive moxibustion sensation. Although both types are based on patients' feeling, positioning method of heat-sensitive moxibustion stands on a new angle and uses a new method to locate acupoint. Therefore, it is believed that the positioning method of heat-sensitive moxibustion is the inheritance and development of ashi method.
Acupuncture Points ; China ; History, Ancient ; Humans ; Medicine in Literature ; Moxibustion ; history ; methods ; Sensation

OBJECTIVETo observe clinical therapeutic effects of acupuncture for pattern of liver-stagnation and spleen-deficiency in diarrhea-predominant irritable bowel syndrome (D-IBS) and its impact on cell factors.

METHODSForty cases were selected and divided into an acupuncture group (21 cases) in which acupuncture was applied and a medicine group (19 cases) in which oral administration of dicetel and bifidobacterium lactobacillus triple viable capsules were applied. The symptom scores, level of Th1-type cytokine (IFN-gamma, IL-2) and Th2-type cytokine (IL-4, IL-10) and ratio of IFN-gamma to IL-4 were compared in two groups before and after treatment to analyze acupuncture effect.

RESULTSThe clinical symptoms were improved after one-week treatment in the acupuncture group (P<0.05), which had faster onset than the medicine group (P<0.05). The total effective rate was 90.48% (19/21) in the acupuncture group, which was superior to 78.95% (15/19) in the medicine group (P<0.05). Compared with medicine treatment, imbalanced condition of Th1/Th2 was turning towards the direction of Th2 after acupuncture, indicating a tendency to recover the balance.

CONCLUSIONThe clinical efficacy of acupuncture for D-IBS has close relationship with effectively improving balance of Th1/Th2 in patients with liver-stagnation and spleen-deficiency.

Acupuncture Therapy ; Adult ; Aged ; Cytokines ; immunology ; Female ; Humans ; Irritable Bowel Syndrome ; immunology ; physiopathology ; therapy ; Liver ; physiopathology ; Male ; Middle Aged ; Spleen ; physiopathology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Young Adult

OBJECTIVESTo assess bone marrow morphologic changes in Philadelphia-chromosome positive chronic myeloid leukemia (Ph(+)-CML) patients treated with Imatinib, and to evaluate the correlation of the morphologic changes with hematological or cytogenetic responses.

METHODSOne hundred and seventeen patients with Ph(+) CML: 54 in chronic phase but failed to interferon-alpha treatment, 41 in accelerated phase, 22 in blastic phase received oral administration of Imatinib 400 or 600 mg once daily for more than 18 months.

RESULTSAll of the patients responded to the treatment, including complete hematological response, bone marrow response and return to chronic phase, bone marrow cellularity and myeloblast count reduced significantly to non-CML picture. Myeloid/erythroid ratio and megkaryocyte count were decreased significantly in most patients in chronic and accelerated phases (P < 0.05). Bone marrow hypoplasia or aplasia was associated with lower cytogenetic response rates in patients in chronic phase (58.8% vs 86.5%, P = 0.035), lower complete hematological response in patients in accelerated phase (26.3% vs 75.0%, P = 0.004), and 6-month overall survival in patients in blastic phase (77.8% vs 16.7%, P = 0.009). Patients in advanced stage obtained non-CML marrow picture in 1 month of treatment had better prognosis. 18-month disease progression rates were lower (25% vs 75%, P = 0.028) and overall survival rates higher (75.0% vs 11.8%, P = 0.004) in patients obtained non-CML picture marrows than in those with CML marrows picture in accelerated phase. Hematological response rate and overall survival of more than 6 months were higher in patients with non-CML marrows picture than those with CML marrows picture (100.0% vs 40.0%, P = 0.017 and 83.3% vs 26.7%, P = 0.046 respectively) in blastic phase.

CONCLUSIONSNormal marrow appearance can be sustained under continuous treatment of Imatinib in CML patients who achieved hematological responses.

Adolescent ; Adult ; Aged ; Benzamides ; Bone Marrow Cells ; cytology ; drug effects ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; pathology ; Male ; Middle Aged ; Piperazines ; administration & dosage ; pharmacology ; Pyrimidines ; administration & dosage ; pharmacology ; Treatment Outcome ; Young Adult

OBJECTIVETo identify and analyse the different species, same species in different regions and confusion species.

METHODNear-infrared diffuse reflectance spectrometry was used.

RESULTClustering analysis showed that clustering relations were far among different Gryllotalpa species and close among the same species from different regions, and there were close relations among the same species from near regions and between Teleogryllus emmus and G. orientalis.

CONCLUSIONNear-infrared diffuse reflectance spectrometry method can be used in classification and identification of Gryllotalpa.

Animals ; Cluster Analysis ; Drug Contamination ; Gryllidae ; chemistry ; classification ; Materia Medica ; chemistry ; classification ; Pharmacognosy ; Species Specificity ; Spectroscopy, Near-Infrared

Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.

OBJECTIVETo investigate the role of p38 mitogen-activated protein kinase (MAPK) signal transduction pathway in the Kupffer cells production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in severely burns rats.

METHODSMale health adult Sprague-Dawley rats were randomized into four groups: sham burn rats given vehicle, sham burn rats given the p38 MAP kinase inhibitor SB203580, rats given a 30% total body surface area (TBSA) full-thickness burn and fluid resuscitation plus vehicle, and burn rats given injury and fluid resuscitation plus SB203580. Rats from each group were killed at 24 h after burn or sham burn and Kupffer cells (KCs) were isolated. After 18 h incubation, KCs next were stimulated with 50 ng/ml of LPS for 18 h. After stimulation, supernatants were removed for analysis of TNF-alpha and IL-1beta levels by ELISA. The TNF-alpha and IL-1beta mRNA expressions (by quantitative real-time RT-PCR) and the activities of p38 MAPK and JNK (by Western blot analysis) in KCs were examined.

RESULTSEighteen hours after 50 ng/ml LPS stimulation, KCs from burn rats released significantly higher levels of TNF-alpha and IL-1beta than did shams. The mRNA levels of TNF-alpha and IL-1beta in KCs increased significantly postburn. Western blot analysis suggested that expression of phosphorylated p38 MAPK and JNK were increased in KCs harvested from burn group after stimulation with LPS compared with those from sham group. In vivo administration of SB203580 markedly suppressed both the release of TNF-alpha and IL-1beta and the mRNA expressions of TNF-alpha and IL-1beta in KCs from both sham and burn rats. p38 MAPK activity in KCs was abolished by administration with SB203580, whereas JNK was not.

CONCLUSIONSp38 MAPK signal transduction pathway mediates KCs production of proinflammatory cytokines TNF-alpha and IL-1beta in severely burned rats.

Animals ; Blotting, Western ; Burns ; enzymology ; physiopathology ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Interleukin-1 ; genetics ; metabolism ; Kupffer Cells ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology

OBJECTIVETo study the effect of multiglycosides of Tripterygium wilfordii (MTW) for treatment of proteinuria in kidney transplant recipients.

METHODForty-five kidney transplant recipients with proternuria were randomized into 3 groups (n=15) and received full daily dose (1 mg/kg) MTW, half dose (0.5 mg/kg) MTW or no MTW (control) in addition to immunosuppressant therapy. The 24-hour urinary protein (24 h Upro), blood urea nitrogen (BUN), serum creatinine (Scr), dose of ciclosporin and the adverse effects of MTW were recorded.

RESULTSMTW at both the full dose and half dose significantly reduced the 24 h Upro as compared to exclusive immunosuppressant therapy (P<0.05). The therapeutic dose of ciclosporin in patients with full and half dose of MTW was significantly lower than that in the control group (P<0.05), and the patients receiving full dose MTW showed greater adverse effects than those having half dose MTW (P<0.05).

CONCLUSIONSMTW can significantly ameliorate proteinuria, reduce the therapeutic dose of ciclosporin and protect the renal function in kidney transplant recipients. While producing similar therapeutic effect to routine full dose, long-term use of half dose MTW may reduce the adverse effect associated with MTW.

Adult ; Aged ; Female ; Glycosides ; therapeutic use ; Graft Survival ; immunology ; Humans ; Kidney Transplantation ; adverse effects ; immunology ; Male ; Middle Aged ; Proteinuria ; drug therapy ; etiology ; Tripterygium ; chemistry ; Young Adult

OBJECTIVETo investigate the effect of fosinopril (Fos) on regulating klotho gene expression and elucidate the mechanism of Fos regulating the Angiotensin II (AngII) -induced down-expression of klotho gene.

METHODSCulture cells, NRK-52E, were incubated with media either AngII or Fos or both of all. Experimental groups incubated with Fos (10(-5) mol/L) were divided according to variant points of time for 0 (control), 3, 6, 12, 24 h. Different concentration of Fos was selected to incubated with culture cells for 0 (control), 10(-9) 10(-8), 10(-7), 10(-6), 10(-5) mol/L at the optimal time point (24 h). Five groups, which were A: control; B: AngII (10(-7) mol/L); C: Fos(10(-5) mol/L); D: AngII (10(-7) mol/L) + Fos(10(-5) mol/L) and E: Cells pretreated with Fos(10(-5) mol/L)12 h incubated with AngII (10(-7) mol/L) were divided to observe the effect of Fos on expression of klotho induced by AngII. RT-PCR and immunohistochemistry (IHC) were applied to evaluate the klotho mRNA and protein expression, respectively.

RESULTSFos up-regulated klotho mRNA in time-dependent manner, and independent of dose-dependent manner; AngII obviously decreased the levels of kloltho mRNA and protein expression in NRK-52E as compared to the control (P < 0.05), the down-regulating effect was reversed by incubating both with AngII and Fos (P < 0.05), and Fos could inhibit the down-regulated expression of klotho gene induced by Ang II in NRK-52E.

CONCLUSIONFosinopril up-regulates klotho mRNA in time-dependent manner, and inhibits the down-regulated expression of klotho gene induced by Ang II.

Angiotensin II ; pharmacology ; Animals ; Cells, Cultured ; Down-Regulation ; Epithelial Cells ; drug effects ; metabolism ; Fosinopril ; pharmacology ; Gene Expression ; drug effects ; Glucuronidase ; genetics ; metabolism ; Kidney Tubules ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Rats