The antifungal antibiotic produced by Streptomyces luteogriseus H-103 was purified by means of macroporous adsorbent resin, and the crystal of the antibiotic with high purity was got. In this paper, the methods of purification by adsorbing of microporous adsorbent resin and detection by reversed high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) were established. The result appeared that resin X-5 is the best adsorbent, the eluant is 50% ethanol. The antibiotic was successfully separated on Agilent~(TM)20RBA?310SB C_(18 )column (150mm?4.6mm i.d,5?m) , using a mixture of acetonitrile (A)-H_(2)O (B) as a mobile phase under gradient elution at a flow of 0.8mL/min at 30℃.0~4.0 min, V(A)∶V(B)=20∶80, 4.0~9.5min, V(A)∶V(B)=45∶55, then V(A)∶V(B)=80∶20. The drift tube temperature and the air carrier gas flow rate of the ELSD were set at 115℃ and 2.3L/min.

Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Nucleosides ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; Pyridazines ; administration & dosage ; pharmacology ; Stomach Neoplasms ; pathology

OBJECTIVETo observe the effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.

METHODThirty-two NOD mice were randomly divided into the model group, the traditional Chinese medicine group (TCM group, orally given 0.4 mL nourishing Yin, strengthening Qi and activating blood decoction as per 100 g x kg(-1) everyday), the hydroxychloroquine group (given 0.4 mL hydroxychloroquine as per 60 mg x kg(-1) everyday), the traditional Chinese medicine and western medicine group (TCM WM group, given nourishing Yin, Strengthening Qi and activating blood decoction 50 g x kg(-1) and hydroxychloroquine 60 mg x kg(-1), 0.4 mL everyday), with eight mice in each group. Eight Balb/C mice were selected as the normal control group (normal group). All of mice were killed after eight weeks, and their submaxillary glands were dissected. The expression levels of Fas/FasL were examined by immunohistochemical method, and the FasL mRNA was detected by RT-PCR.

RESULTThe expression levels of Fas/FasL in salivary glands of the model group were higher than that of other groups (P < 0.05). The expression level of FasL of the normal group was much lower than that in the hydroxychloroquine group (P < 0.05). The relative expression level of Fas mRNA in salivary glands of the model group was higher than that in other groups, but the control group was notably lower than other groups (P < 0.05). The expression level of FasL mRNA in salivary glands of the model group was higher than that in TCM and TCM WM groups (P < 0.05). But the expression level in TCM WM group was notably lower than the hydroxychloroquine group (P < 0.05).

CONCLUSIONThe nourishing Yin, strengthening Qi and activating blood decoction could down-regulate the expression level of Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression, and had a better efficacy after being combined with hydroxychloroquine. The nourishing Yin, strengthening Qi and activating blood decoction might treat the Sjogren's Syndrome by reducing apoptosis which is regulated by Fas/FasL

Animals ; Fas Ligand Protein ; genetics ; Female ; Gene Expression Regulation ; Medicine, Chinese Traditional ; methods ; Mice ; Mice, Inbred NOD ; Qi ; RNA, Messenger ; genetics ; metabolism ; Salivary Glands ; metabolism ; Sjogren's Syndrome ; blood ; genetics ; therapy ; Yin-Yang ; fas Receptor ; genetics

How to get functional gene from uncultured-microbiology is the hotspot content of microbial ecology. What the most important is how to obtain the pure and integrated genomic DNA. An efficient, nonselective extraction method to gain chromosomal DNA from eight kinds of bacteria was introduced. Amount DNA released by hot-detergent gave the highest DNA yields from different G + and G- bacteria. Running 20 hours by PFGE mode, the size of total DNA is over 23kb. The pure DNA could be digested by Hind Ⅲ and used in PCR. The total environmental DNA also can be extracted from soil by the same method. As a result it showed a new way for the environmental DNA extraction.

Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.

OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

Objective: To observe the clinical efficacy of sinew-bone balancing manipulation in treating lumbar disc herniation (LDH) and offer clinical evidence to support the concept of paying equal attention to sinew and bone. Methods: Sixty LDH patients were randomized into an observation group and a control group, with 30 cases in each group. The observation group was treated with the sinew-bone balancing manipulation, and the control group received conventional Tuina (Chinese therapeutic massage) manipulation. The clinical efficacy and posterior muscle chain tone effect were compared between the two groups by observing the visual analog scale (VAS) score, Japanese Orthopaedic Association (JOA) score, and posture-associated indicators. Results: The total effective rate was 86.7％ in the observation group, higher than 76.7％ in the control group, but the between-group difference in efficacy was statistically insignificant (P>0.05). After treatment, the VAS and JOA scores, angle-dependent muscle tone indicator of the posterior muscle chain, and lumbar posture symmetry showed significant changes in both groups (P<0.05). The VAS and JOA scores, angle-dependent muscle tone indicator of the posterior muscle chain, and lumbar posture symmetry in the observation group were significantly different from those in the control group after treatment (P<0.05), but the between-group difference in the general posture symmetry was statistically insignificant (P>0.05). Conclusion: Both the sinew-bone balancing and conventional Tuina manipulations can reduce pain in LDH patients, improve lumbar function, and adjust the angle-dependent muscle tone coefficient of the posterior muscle chain and lumbar posture symmetry; except for the general posture symmetry of the posterior muscle chain, the sinew-bone balancing manipulation wins out over the conventional Tuina manipulation.

OBJECTIVETo observe the effect of Yangyin Jiedu Huoxue Recipe (YJHR) on the degree of activity integral of systemic lupus erythematosus (SLEDAI) and plasma osteopontin (OPN), and its effect for hormone withdrawal in patients with systemic lupus erythematosus (SLE).

METHODSSeventy-eight patients were randomly assigned by a randomizing digital table to two groups, 42 patients in the treated group and 36 in the control group, all were treated with conventional Western medical treatment, and YJHR was given additionally to the treated group. Changes of SLEDAI and OPN, as well as the daily dosage of prednisone used were observed after 2 courses of treatment.

RESULTSSLEDAI and OPN were lowered in both groups, but the lowering in the treated group [9.17 +/- 4.12, (117.69 +/- 78.50) microg/L] was more significant than that in the control group [11.60 +/- 4.05, (151.09 +/- 83.90) microg/L, P<0.05]. The two indices were positively correlated. The dosage of prednisone daily used in the treated group (10.03 +/- 4.29) was reduced more than that in the control group (15.14 +/- 6.67, P<0.05). SLEDAI of all the patients before and after treatment had a positive relationship with OPN respectively (r=0.44, 0.39) (P<0.05).

CONCLUSIONSYJHR can suppress the activity of disease and be helpful for early withdrawal of prednisone in treating SLE. Plasma level of OPN is related with the activity of the disease.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lupus Erythematosus, Systemic ; blood ; drug therapy ; Middle Aged ; Osteopontin ; blood ; Phytotherapy ; Prednisone ; therapeutic use ; Young Adult

OBJECTIVETo investigate the immuno-regulatory and therapeutic effects of Chinese herbal medicine for Yiqi Yangyin Quyu (YYQ, supplementing qi, nourishing yin, removing blood stasis) on Sjögren's syndrome (SS).

METHODSSixty-two patients were randomly divided into the treated group (37 cases) treated with Chinese medicine and prednisone, and the control group (25 cases) treated with prednisone alone, and another 20 healthy persons were taken as the normal control group. Curative effects was observed, and immunoglobulin (Ig) and T lymphocyte subsets were detected before and after 3 months' treatment.

RESULTSImmunoglobulin (IgG, IgM and IgA) before treatment in SS patients were higher, while the proportion of natural killer (NK) cell and CD4 were lower as compared with those in the normal control group. After treatment, IgG, IgM and IgA lowered to normal levels (P < 0.05), the proportion of NK cell and CD4 rose in both groups (P < 0.05), but CD3 in the control group was still lower than that in the treated group (P < 0.05). The total curative effective rate in the treated group was 91.9%, better than that in the control group (76.0%, chi-squared = 3.92, P < 0.05).

CONCLUSIONChinese herbal medicine for YYQ could improve clinical curative effective rate and regulate immune function in SS patients.

Adult ; Aged ; Anti-Inflammatory Agents ; therapeutic use ; CD4 Antigens ; immunology ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulins ; immunology ; Killer Cells, Natural ; drug effects ; immunology ; Male ; Middle Aged ; Phytotherapy ; Prednisone ; therapeutic use ; Sjogren's Syndrome ; drug therapy ; immunology ; Yin Deficiency ; drug therapy ; immunology

BACKGROUNDSleep/wake disturbances in patients with amyotrophic lateral sclerosis (ALS) are well-documented, however, no animal or mechanistic studies on these disturbances exist. Orexin is a crucial neurotransmitter in promoting wakefulness in sleep/wake regulation, and may play an important role in sleep disturbances in ALS. In this study, we used SOD1-G93A transgenic mice as an ALS mouse model to investigate the sleep/wake disturbances and their possible mechanisms in ALS.

METHODSElectroencephalogram/electromyogram recordings were performed in SOD1-G93A transgenic mice and their littermate control mice at the ages of 90 and 120 days, and the samples obtained from these groups were subjected to quantitative reverse transcriptase-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay.

RESULTSFor the first time in SOD1-G93A transgenic mice, we observed significantly increased wakefulness, reduced sleep time, and up-regulated orexins (prepro-orexin, orexin A and B) at both 90 and 120 days. Correlation analysis confirmed moderate to high correlations between sleep/wake time (total sleep time, wakefulness time, rapid eye movement [REM] sleep time, non-REM sleep time, and deep sleep time) and increase in orexins (prepro-orexin, orexin A and B).

CONCLUSIONSleep/wake disturbances occur before disease onset in this ALS mouse model. Increased orexins may promote wakefulness and result in these disturbances before and after disease onset, thus making them potential therapeutic targets for amelioration of sleep disturbances in ALS. Further studies are required to elucidate the underlying mechanisms in the future.

Amyotrophic Lateral Sclerosis ; genetics ; metabolism ; Animals ; Female ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Transgenic ; Neuropeptides ; genetics ; metabolism ; Orexins ; Reverse Transcriptase Polymerase Chain Reaction ; Sleep ; physiology ; Superoxide Dismutase ; genetics ; metabolism ; Superoxide Dismutase-1 ; Wakefulness ; physiology