Objective To construct luciferase reporter plasmids of truncated fragments of different lengths of human guanylate binding protein 5(GBP5)gene promoter and analyze the transcriptional activity of each fragment to determine the core regulatory region.Methods GBP5promoter sequence was amplified by PCR,truncated into five fragments of different lengths and connected to pGL3-basic plasmid.The constructed recombinant plasmids pGL3-GBP5-11/21/31/41/51were transfected into 293FT cells and detected for luciferase activity.The binding sites of transcription factors in GBP5promoter region were predicted by JASPAR software,and Yin-Yang transcription factor 1(YY1)targeting the core regulatory region was selected and verified for the transcriptional regulatory activity.The CDS sequence of YY1 was amplified by PCR to construct the overexpression plasmid pIRES2-EGFP-YY1,which was then co-transfected to 293FT cells with plasmids pGL3-GBP5-21(-1 623 ～ +47 bp)and internal reference plasmid pRL-CMV,and detected for luciferase activity to analyze the regulation of transcription factor YY1 on GBP5 promoter activity.Results Colony PCR and double enzyme digestion identification proved that the plasmid of human GBP5 promoter reporter gene was correctly constructed;JASPAR software predicted that there were multiple transcription factor binding sites such as STAT1,YY1 and Foxp3 in GBP5promoter region.Double luciferase activity assay showed that pGL3-GBP5-21(-1 623 ～ +47 bp)showed the highest promoter activity,while the promoter activity of pGL3-GBP5-41(-520 ～ +47 bp)decreased significantly,suggesting that the core region of GBP5 promoter was located at upstream-1 623 ～-520 bp of 5 'UTR;Overexpression of YY1 significantly activated the GBP5 promoter activity and regulated the expression of GBP5.Conclusion The core regulatory region of human GBP5 promoter was located in upstream-1 623 ～-520 bp of the 5 'UTR,with a binding site of transcription factor YY1 existing in this region.Meanwhile,overexpression of YY1 significantly effected the activity of GBP5 promoter.

OBJECTIVETo investigate extracellular splitting pattern of mitochondria and the depressant effects of CsA on the process and explore the mechanism of post-traumatic SIRS and its therapeutic strategy.

METHODSTen male SD rats with 60 to 70 days age and 240 to 280 g weight were used for mitochondrial isolation. Freshly isolated mitochondria were randomly divided into two groups, which were cultured in blood plasma with or without CsA respectively for 8 h. COX and MDH were assayed by ELISA every 30 min. Meanwhile, Rat macrophage cell line NR8383 were treated as follows, control (group A): cultivation with normal medium; NR8383+CsA co-culture group (group B): culture medium was supplemented with CsA of 10 mmol/L; NR8383+intact mitochondria co-culture group (group C): culture medium was supplemented with intact mitochondria (mtDNA=5 g/ml); NR8383+intact mitochondria+CsA co-culture group (group D): culture medium was supplemented with intact mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L; NR8383+disrupted mitochondria co-culture group (group E): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml); NR8383+disrupted mitochondria+CsA co-culture group (group F): culture medium was supplemented with disrupted mitochondria (mtDNA=5 μg/ml)and CsA of 10 mmol/L. TNF-α and IL-6 concentrations in supernatant were assessed at 1, 3, 5 h after culture.

RESULTSIn the mitochondria plasma cultures, MDH and COX levels were increased with the time and peaked at about 3 h and 3.5 h; CsA can delay the appearance of peak to 4.5 h. Among different treated groups,there was no significant difference in TNF-α and IL-6 between group A and group B; there was significant difference in TNF-α and IL-6 other groups. After 1 h culture, compared with group C, no significant difference of TNF-α and IL-6 was observed in group D, while TNF-α and IL-6 were significant higher in group E; after 3 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while TNF-α and IL-6 were significantly higher in group E; after 5 h culture, compared with group C, TNF-α and IL-6 were significantly lower in group D, while no significant difference of TNF-α and IL-6 were observed in group E. At each time point, there was no significant difference in TNF-α and IL-6 between group F and group E.

CONCLUSIONMitochondria can split in serum with time, which will further activate macrophages. CsA has depressant effect to mitochondrial splitting on the process and will therefore inhibit the activation of macrophages.

Animals ; Cells, Cultured ; Cyclosporine ; pharmacology ; Interleukin-6 ; secretion ; Male ; Mitochondria ; drug effects ; Prostaglandin-Endoperoxide Synthases ; analysis ; Rats ; Rats, Sprague-Dawley ; Systemic Inflammatory Response Syndrome ; drug therapy ; etiology ; Tumor Necrosis Factor-alpha ; secretion

OBJECTIVETo observe interventional effects of anti-viral therapy and Compound Qin-gre Granule (CQG) on host cellular immune functions of acute virus infection patients.

METHODSThirty acute virus infection patients were recruited to detect peripheral lymphocyte subsets. They were randomly assigned to two groups, the Western medicine treatment group (treated with anti-virus Western medicine) and the integrative medicine treatment group (treated with anti-virus Western medicine plus CQG). T-cell subsets were re-examined 7 days later. Changes between before and after treatment were observed. Effect on host cellular immune functions and efficacy were compared between the Western medicine treatment and the integrative medicine treatment.

RESULTSCompared with the normal control group, the percentage of peripheral T cells increased, and the percentage of B/NK cells decreased in acute virus infection patients (P < 0.01). Meanwhile, in T cell subsets, the percentage of CD8+ T cells and CD8+ CD38+ T cells increased (P < 0.05, P < 0.01); and percentages of CD4+ T cells, CD4+ CD28 + T cells, and CD8+ CD28+ T cells decreased (P < 0.05, P < 0.01). After one-week treatment, percentages of CD4+ T cells, CD4+ CD28+ T cells, and CD8+ CD28+ T cells increased (P < 0.05, P < 0.01), while the percentage of CD8+ CD38+ T cells decreased (P < 0.01). More significantly, these changes were greater in the integrative medicine treatment group than in the Western medicine treatment group (P < 0.05).

CONCLUSIONSDisarranged cellular immune functions existed in acute virus infection patients. CQG could significantly improve viral infection induced immunologic derangement and immunologic injury.

Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lymphocyte Count ; Lymphocyte Subsets ; T-Lymphocyte Subsets ; Virus Diseases ; drug therapy

OBJECTIVETo explore the expression of p-p38 MAPK protein and the number of astrocytes expressing p-p38 MAPK in CA1 hippocampus in rats during the induction of brain ischemic tolerance induced by intermittent hypobaric hypoxia (IH) preconditioning.

METHODSThirty healthy adult male Wistar rats were randomly divided into 6 groups (n = 5 in each group): sham 0 min group, IH + sham 0 min group, sham 7 d group, IH + sham 7 d group, Ischemia (Is) 7 d group, and IH + Is 7 d group. Neuropathological evaluation was performed by thionine staining in CA1 hippocampus in rats. The expression of p-p38 MAPK in CA1 hippocampus was observed by immunohistochemical staining. And the number of astrocytes expressing p-p38 MAPK was observed by immunofluorescent double labeling.

RESULTSThe results showed that IH preconditioning induced brain ischemic tolerance successfully. At the same time, IH preconditioning obviously up-regulated the expression of p-p38 MAPK protein in CA1 hippocampus, and also increased the number of astrocytes expressing p-p38 MAPK.

CONCLUSIONIt might be concluded that IH preconditioning induced brain ischemic tolerance by up-regulating the expression of p-p38 MAPK protein in pyramidal neurones and astrocytes.

Animals ; Astrocytes ; enzymology ; pathology ; Brain Ischemia ; enzymology ; pathology ; Disease Models, Animal ; Hippocampus ; enzymology ; Hypoxia ; Ischemic Preconditioning ; methods ; Male ; Phosphorylation ; Pressure ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; metabolism

The water-soluble polysaccharide PNM isolated from the submerged culture of Phellinus nigiricans by hot water extraction and ethanol precipitation, was purified by freeze thawing, methods of Sevag, and then fractionated by Sepharose CL-6B gel filtration chromatography, giving a polysaccharide fraction named as PNMⅠ. HPLC analysis showed that PNMⅠwas a homogeneous polysaccharide, with a weight-average molecular weight of 29 kDa. GC analysis indicated that PNMⅠis a heteropolysaccharide composed of mannose, galactose and glucose with a ratio of 0.35:0.44:1.00. Pharmaceutical experiments showed both PNM and PNMⅠ have a significant effect on the immunomodulatory activities of murine splenic lymphocytes in vitro. Also PNMⅠ could inhibit the growth of sarcoma 180 solid tumor in vivo.

The effects of four kinds of carbon sources and nine kinds of nitrogen sources on the fungus Phellinus nigricans mycelia were studied.The four kinds of carbon source are sucrose,lactose,glucose,soluble starch.The nine kinds of nitrogen source are corn meal,wheat bran,potato,soybean meal,yeast extract powder,peptone,potassium nitrate,ammonium nitrate,and urea.Polysaccharides were extracted from different generations of mycelia and the contents,monosaccharide components and molecular weight were analyzed.The results were that the optical carbon source and nitrogen source were starch and corn meal.The optical combination was sucrose and corn meal.The properties of polysaccharides from different generations of mycelia were stable.

OBJECTIVE To discuss the clinical traits,pathogenesis and traditional Chinese medicine(TCM) stepwise treatments of the patients who suffered from pulmonary infection after kidney transplantation.METHODS Because of patients after kidney transplantation easily suffered from pulmonary infection,which was characterized by non-typical symptoms,concealment and varity.In the aspects of the pathogenesis of this disease,the principal aspect of which was the weakened body resistance(kidney deficiency)and the secondary incidental was affected pathogenic factor(heat,toxin,stasis brewing in the lungs),viz weaken healthy qi and excessive pathogenic factor.Hence during the clinical treatment we should attach importance to regulate entire faculty condition.Considering the acute attack stage and restoration stage,we would differently inflict therapies of clearing heat,resolving toxin,quickening the blood,supplementing the kidney and nourishing yin,clearing heat and moistening the lung and so on,according to differentiation of symptoms and signs,in which way we could give attention to both the root and tip.RESULTS The TCM stepwise therapy of pulmonary infection after kidney transplantation had the characteristics of high efficacy,few side reactions and stable long-term curative effects.CONCLUSIONS The TCM therapy of pulmonary infection after kidney transplantation has more potentiality and predominance,which deserves further study.

OBJECTIVE To discuss the clinical traits,pathogenesis and TCM stepwise treatments of fungal urinary tract infection.METHODS According to the risk factors and clinical character of fungal urinary tract infection,we clarified the mechanism of the disease.The principal aspect was spleen-kidney vacuity detriment and the secondary incidental was accumulated damp-heat and static blood in the lower burner,viz weaken healthy qi and excessive pathogenic factor.Hence during the clinical treatment we should regulate faculty condition.In the acute infection period we should give priority to dispel evils supplemented by the recovery of right qi.In the convalescence we should pay more attention to support right supplemented by dispelling.RESULTS The most common pathogen of fungal urinary tract infection was Candida albicans.The TCM stepwise treatments of fungal urinary tract infection together with regulatiy entire faculty condition had the characteristics of high efficacy and few side effects.CONCLUSIONS The TCM stepwise treatments of fungal urinary tract infection has more potentiality which deserves further study.

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon, Coronary ; Combined Modality Therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Myocardial Infarction ; drug therapy ; therapy ; Phytotherapy ; Stents