Altitude ; Animals ; Arvicolinae ; Gene Expression ; Hypoxia ; Rats ; Tumor Suppressor Protein p53 ; genetics

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

OBJECTIVETo explore the therapeutic effect of qi benefiting blood activating method (QB-BAM) on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients with blood stasis syndrome (BSS) by observing its effect on plasma fibrinogen (Fg) and D-dimer (D-D) levels.

METHODSSixty AECOPD patients with BSS were randomly assigned to the treated group and the control group, 30 in each group. All patients received conventional therapy for AECOPD. Those in the treated group were additionally injected with Shengmai Injection and Tanshinone IIA Injection. Clinical efficacy and indices including levels of Fg, D-D, PaO2, and PaCO2 were measured and compared before and after treatment.

RESULTSThe effective rate was 93.3% (28/30 cases) in the treated group, higher than that of the control group [73.3% (22/30 cases) , P < 0.05]. There was no significant difference in all indices between the treated group and the control group before treatment (P >0.05). After treatment all indices were significantly improved in the two groups (P < 0.01). But in the treated group levels of Fg and D-D decreased more and levels of PaO2 increased more (P < 0.01). Plasma levels of Fg and D-D levels were negatively correlated with PaO2 (r = -0.493, r = -0.438, P < 0.01) before treatment, and also negatively correlated with PaO2 (r = -0.452, r = -0.325, P < 0.01, P < 0.05) after treatment, but they were not significantly correlated with PaCO2 before and after treatment (P >0.05).

CONCLUSIONSQBBAM could play a therapeutic role in improving prethrombotic states of AECOPD patients with BSS. Plasma levels of Fg and D-D were related to the severity of AECOPD.

Acute Disease ; Drugs, Chinese Herbal ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; Fibrinogen ; Hemostatics ; Humans ; Plasma ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Qi

OBJECTIVETo investigate the effect of the mammalian target of rapamycin (mTOR) inhibitor CCI-779 on the chemosensitivity of androgen-independent prostate cancer cell line PC-3.

METHODSProstate cancer cells PC-3 were cultured and treated with CCI-779, Paclitaxel and combination of the two. Then the inhibitory effects of the three medications on the growth of the PC-3 cells were determined by MTT, and the their cell cycle and apoptosis were detected by flow cytometry.

RESULTSCompared with the control group, the three medications all significantly inhibited the proliferation of the PC-3 cells, and the combined method even enhanced the effect. Flow cytometry showed that CCI-779 and Paclitaxel blocked the cell cycle mainly in the G1/G2 stage, while the combined medication mainly in the G0/G1 stage. Significantly increased apoptosis of the PC-3 cells was observed in the three medication groups as compared with the control group (P < 0.01).

CONCLUSIONCCI-779 can inhibit the proliferation of PC-3 cells and enhance the chemosensitivity of prostate cancer.

Antineoplastic Agents ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drug Therapy, Combination ; Humans ; Male ; Paclitaxel ; pharmacology ; Prostatic Neoplasms ; drug therapy ; Protein Kinase Inhibitors ; pharmacology ; Sirolimus ; analogs & derivatives ; antagonists & inhibitors ; pharmacology

AIMTo separate and purify the anti-myocardial ischemic polysaccharide fraction with a homogenous molecular weight from Ophiopogon japonicus, then study the chemical structure of the parts.

METHODSCrude polysaccharides were prepared by extracting the tube root fraction of Ophiopogon japonicus with water, then precipitation with ethanol. From the crude polysaccharides, the polysaccharide of MDG-1 was separated and purified using ultrafiltration, DEAE Sepharose FF and Sephadex G-25 column chromatography. Its structure was studied by complete hydrolysis, periodate oxidation, Smith degradation, methylation analysis, 1H NMR and 13C NMR analysis etc.

RESULTS AND CONCLUSIONMDG-1 was a water-soluble beta-D-fructosan, containing a backbone composed of Fruf (2 --> 1), and a branch of Fruf (2 --> 6) Fruf (2 --> per average 2. 8 of main chain residues. Mn, Mw and Mp of MDG-1 were 3 400, 4 800 and 5 000, respectively. MDG-1 contains trace of Glc, which maybe connect to its reducing terminal. Molar ratio of Fru and Glc is approximately 35: 1.

Methylation ; Molecular Structure ; Molecular Weight ; Ophiopogon ; chemistry ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; isolation & purification

The loquat is widely cultivated in China, its succulent fruits, leaves and flower are used as a traditional medicine for the treatment of many diseases. The study is aimed to analyse the content of the four triterpene compounds ( ursolic acid, corosolic acid, maslinic acid, oleanolic acid) in different organs, and investigate the dynamic changes in different phenological period. The triterpenic acids content in the samples was measured by HPLC based on the plant phenological observations. The results showed that order of four triterpenic acids content in different organs from high to low was defoliation (23.2 mg x g(-1)) > mature leaves (21.7 mg x g(-1)) > young leaves (17.5 mg x g(-1)) > fruits (7.36 mg x g(-1)) > flowers (6.40 mg x g(-1)). The triterpenic acids were not detected in the seeds. The total amount of the four triterpenic acids in the loquat leaves collected in the different phenological stages of sprout, flower bud, blossom and fruit varied between 17.8 and 26.2 mg x g(-1) (defoliation), 16.5 and 23.5 mg x g(-1) (mature leaves), 14.7 and 21.5 mg x g(-1) (young leaves), respectively. The content increased progressively with the leaf development, maturation and aging. There was a higher level of the dry material and triterpenic acids accumulation in the mature leaves during fruit enlargement. This paper attempts to present the case for medicinal plants of a broad geographical distribution to study on the secondary metabolites and harvesting time.
China ; Chromatography, High Pressure Liquid ; Eriobotrya ; chemistry ; growth & development ; Flowers ; chemistry ; growth & development ; Fruit ; chemistry ; growth & development ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; Seeds ; chemistry ; growth & development ; Triterpenes ; analysis

OBJECTIVETo determine the effects of diazoxide on oxygen free radicals and cell apoptosis in brain tissue after deep hypothermia cerebral ischemia reperfusion injury in young rats.

METHODSFifty-four 3-week-old Sprague-Dawley rats were randomly and equitably divided into sham-operated group, model group and diazoxide group respectively (n = 18). The model of hypothermia cerebral ischemia reperfusion injury was made. After 24 hours of operation, the brains of rats were removed and preserved. The content of superoxide dismutase (SOD) and malonaldehyde (MDA) in brain tissue were detected. Cytosolic C release of cytochrome was confirmed by Western Blot. The protein expression of Caspase-3 was determined by immunohistochemistry.

RESULTSIn the model group, the content of SOD was (198 +/- 41) U/mg, lower than the sham-operated group's (321 +/- 36) U/mg (P < 0.01). The content of MDA was (212 +/- 21) nmol/mg, was higher than the sham-operated group's (100 +/- 23) nmol/mg (P < 0.01), and the expressions of cytochrome C (0.72 +/- 0.09) and Caspase-3 (83 +/- 10) were all significantly higher than those in the sham-operated group (0.17 +/- 0.02 and 115 +/- 9) (P < 0.01). Compared with the model group, the content of SOD in the diazoxide group [(264 +/- 34) U/mg] was markedly increased (P < 0.05). In addition, diazoxide provided significant reductions in the content of MDA [(174 +/- 19) nmol/mg] and the expressions of cytochrome C (0.41 +/- 0.05) and Caspase-3 (99 +/- 11) (P < 0.05).

CONCLUSIONSThe neuroprotective effects of diazoxide against brain injury induced by deep hypothermia cerebral ischemia reperfusion through inhibiting oxygen free radicals and cell apoptosis. Diazoxide may become a new neuroprotective drug after infant complicated congenital cardiac operation.

Animals ; Apoptosis ; drug effects ; Brain ; metabolism ; pathology ; Brain Ischemia ; etiology ; metabolism ; pathology ; Caspase 3 ; metabolism ; Circulatory Arrest, Deep Hypothermia Induced ; adverse effects ; Cytochromes c ; metabolism ; Diazoxide ; pharmacology ; Disease Models, Animal ; Female ; Male ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reperfusion ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the changes of the contralateral testicular histology and germ cell apoptosis after unilateral testicular torsion (UTT) and to determine whether the contralateral testis is injured or not.

METHODSSixty SD male rats were divided into control group (12 rats) and experimental group(48 rats). The former underwent sham operation of the left testis under general anaesthesia. The latter underwent left testis torsion(720 degrees) for 6 h, and then 4 of them were sacrificed and the other 44 were subdivided into the torsed testis untwisted group (22 rats) and the torsed testis removal group (22 rats), 7-8 rats were sacrificed and both testes (twisted and untwisted) were removed 1 day, 1 week and 4 weeks after surgery. All testes underwent histological and germ cell apoptosis examination.

RESULTSThere were significant histological changes in the contralateral testis, and the germ cell apoptosis was increased greatly in the contralateral testis.

CONCLUSIONSUTT can cause contralateral testicular injury, whose mechanism may be related to reperfusion, and torsed testis removal can prevent or reduce damage to the contralateral testis.

Animals ; Apoptosis ; Germ Cells ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Spermatic Cord Torsion ; pathology ; Testis ; pathology

OBJECTIVETo identify the disease-causing gene in a four-generation Chinese family with 9 members affected with primary congenital lymphoedema (PCL, also known as Milroy disease).

METHODSLinkage analysis was performed with a few microsatellite markers flanking the candidate genetic loci for PCL, including 3 known genes associated with autosomal dominant PCL. For mutation analysis, VEGFR3 gene was sequenced with DNA from the proband. Direct DNA sequencing of exon 25 of the VEGFR3 gene was performed in all family members.

RESULTSThe disease gene in the family was mapped to chromosome 5q35.3 with a maximum Lod score of 2.07. Direct DNA sequencing of VEGFR3 gene revealed a heterozygous C to T transition at nucleotide 3341, resulting in p.Pro1114Leu mutation. The p.Pro1114Leu mutation co-segregated with all affected individuals in the family.

CONCLUSIONThis study identified a C3341T (p.Pro1114Leu) mutation in the VEGFR3 gene in a Chinese family with PCL, provided evidence that VEGFR3 mutation can cause PCL in Chinese.

Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Cataract ; genetics ; Genetic Loci ; Humans ; Lod Score ; Lymphedema ; genetics ; Microsatellite Repeats ; genetics ; Mutation ; Point Mutation ; Vascular Endothelial Growth Factor Receptor-3 ; genetics

To investigate the effects of normal human bone m arrow fibroblastoid stromal cell line (HFCL) on the proliferation of acute myeloid leukemia cell line HL-60 and expression of vascular endothelial growth factor (VEGF), establishing coculture system of leukemia cell line HL-60 and HFCL, growth data was obtained by cell counting. Mitotic index (MI) was observed under Wright-Giemsa staining. Flow cytometry and Western blot were used as assays for cell cycle and expression of proliferating cell nuclear antigen (PCNA) separately. VE GF levels were evaluated by using commercial ELISA kits. The results showed that compared with HL-60 cells without HFCL cells, the proliferation of HL-60 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited. The MI of HL-60 cells without HFCL cells was highest followed by HL-60 cells separated by transwell and HL-60 cells in direct contact with HFCL cells. The expression of PCNA in HL-60 cells with HFCL cells were lower than HL-60 cells without HFCL cells. Meanwhile, the percentage of HL-60 cells in G1 phase cocultured with HFCL cells was higher than that without HFCL cells while the percentage of Sphase cells was lower. The levels of VEGF in HL-60 cells with HFCL cells were lower than that in HL-60 cells alone. In conclusion, the normal bone marrow fibroblastoid stromal cells inhibited the proliferation of HL-60 cells as well as the expression of VEGF.
Cell Cycle ; Cell Division ; Cell Line ; Coculture Techniques ; Fibroblasts ; physiology ; HL-60 Cells ; cytology ; Humans ; Proliferating Cell Nuclear Antigen ; analysis ; Stromal Cells ; physiology ; Vascular Endothelial Growth Factor A ; analysis