Objective To observe the effect of exercise on the expression of TNF-α in the adipose tissue of insulin resistant rats fed a high fat diet. Methods Thirty healthy male rats were randomly divided into a high fat di- et group and a normal chow group. Eighteen weeks later, the high fat group was randomly divided into a resting group fed with the high fat diet only, and an exercise group fed the high fat diet, but receiving swimming training for 6 weeks. Changes in their metabolism of glucose and lipids were observed, and the insulin sensitivity index was calcu-lated. Meanwhile, the level of TNF-α mRNA in their adipose tissue was detected with a real-time fluorescence quan-titative polymerase chain reaction (PCR), and protein in the adipose tissue was measured using Western blotting. Results After 18 weeks of high fat diet feeding, the insulin sensitivity index of the high fat diet group decreased sig-nificantly as compared to the normal chow group, suggesting that insulin resistance had been acquired in the high fat diet group. 24 weeks later, the insulin sensitivity index of the resting group had decreased further, again significantly when compared to the normal chow group. Compared to the resting group, the insulin sensitivity index of exercise group was significantly higher, and the expression of TNF-α mRNA and protein in their adipose tissue was significant- ly increased. Conclusion Insulin resistance can be induced by high fat diet feeding. Exercise can improve insulin resistance by increasing the expression of TNF-α in adipose tissue.

Objective: To investigate the interactions between natural killer (NK) cells and Cryptococcus neoformans(C. neoformans) in patients with cryptococcosis, so as to pave a way for treatment and prevention of cryptococcosis. Methods: The peripheral blood samples of 40 cryptococcosis patients and 40 healthy controls were collected. Expression of CD2, CD3, CD4, CD8, CD28, CD18, CD19, and CD56 in patients peripheral blood mononuclear cells(PBMC) were detected by FACS. Cytotoxic activity of NK cells was analyzed by MTT using K562 cells as target cells and the influence of IFN-α and IL-2 on NK cell activity was also studied. The level of IFN-γ in the culture supernatant was assayed by ELISA and the cytotoxic activity of the supernatant was determined. The transcription levels of perforin, granzyme B and granulysin were examined by quantative real-time PCR. C. neoformans and NK cells were cocultured to investigate the inhibition of NK cells on C. neoformans. Results: Compared with that in healthy controls, CD56+ cells decreased significantly (P<0.01) in the PBMC of patients with cryptococcosis, CD8+,CD19+ and CD18+ cells increased significantly (P<0.05 or P<0.01),and the ratio of CD8+/CD28+ increased and of CD4+/CD8+ declined significantly (P<0.05). However, the levels of CD2+, CD3+ and CD4+ cells had no change. Compared with that in healthy controls, the NK cells cytotoxicity in patients with cryptococcosis decreased significantly(P<0.01), but increased when IFN-α or IL-2 was added (P<0.01). IFN-γ level in the PBMC supernatant of patients with cryptococcosis was much lower than that in healthy controls(P<0.01). Transcription of perforin, granzyme B and granulysin in PBMC of patients with cryptococcosis decreased markedly in contrast to those of healthy controls(P<0.01). The inhibitory effect of NK cells on growth of C. neoformans was significantly lower in cryptococcosis patients than that in healthy controls (P<0.05). Conclusion: The quantity and function of NK cells decrease in patients with cryptococcosis. Cytokines such as IL-2 and IFN-α can upregulate the cytotoxicity of NK cells, which suggests a new way for treatment of cryptococcosis.

Objective: To study the cytotoxic effects of granulysin on Candida albicans. Methods: Candida albicans were cultured with different concen trations of granulysin peptides and the colonies of Candida albicans on Sabouraud dextrose agar were calculated. Broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) of granulysin and fluconazole on Candida albicans. Results: When granulysin peptides 2 and 3 (corresponding to G2 and G3 peptides) were at 20 μg/ml, the average colonies of Candida albicans decreased from 838 and 927 to 203 and 218, respectively; G1, G4 and G5 did not reduce the average colony of Candida albicans even at 40 μg/ml. Three strains of Candida albicans were sensitive to granulysin, with their MICs being 26, 22, 29 μg/ml, and their MICs to fluconazole were 4, 20 and 128 μg/ml. Conclusion: Granulysin has cytotoxic effects on Candida albicans and is one of the natural anti-fungi proteins in human body; it has a promising future for anti-fungi drug development.

Background Sustained abnormal tear secretion in dry eye patients may lead to ocular surface and lacrimal glands in the state of long-term inflammatory cell infiltration.Lacrimal gland suffers immune attack by lymphoproliferative,and inflammation interferes normal gland secretion,in which chemokines and their receptors on lymphocytes play a key role.Objective This study was to investigate the inflammation mechanism of delayed allergy induced by Th1 cell in the development of dry eye.Methods Sixty eyes of 30 patients with dry eye and matched 30 healthy volunteers were enrolled in Affiliated First Hospital of Xinjiang Medical University from June to December in 2012.Schirmer Ⅰ test (S Ⅰ t),break up time (BUT) of tear and corneal fluorescence stain (FLS) were performed on the subjects,and conjunctiva epithelial cells were obtained using cytological method of conjunctiva imprinting.Positive cell rates of CC chemokine receptor 5 (CCR5) and CXC chemokine receptor 3 (CXCR3) were detected by flow cytology,and the relative expression levels of regulated upon activation of normal T cells exp ressed and secreted (RANTES),macrophage inflammatory protein (MIP)-lα and MIP-1β,monokine induced by interferon-γ (IFN-γ) (MIG),interferon-γ inducible protein 10 (IP10) and interferon-inducible T-cell α chemoattractant (I-TAC) mRNA were quantitatively assayed by real-time PCR.Differences of the positive rates of CCR5,CXCR3 and lignds were compared between dry eye group and normal control group.Relationship between the positive rates of CCR5,CXCR3 and BUT,S Ⅰ t,FLS scores was analyzed.Results The values of BUT,S Ⅰ t were (2.90±1.37) seconds and (4.00±2.49) mm/5 minutes in the dry eye group,which were significantly lower than (8.56±4.69) seconds and (11.31 ±5.23) mm/5 minutes in the normal control group (t =3.172,2.186,both at P＜0.05).FLS scores,positive rates of CCR5 and CXCR3 were0.90±0.57,(3.38±0.66) ％ and (2.64±0.47)％ in the dry eye group,showing significant elevations in comparison with 0.14±0.06,(2.12±0.21) ％ and (1.12±0.11) ％ in the normal control group (t=2.297,3.151,5.454,all at P＜0.05).In the dry eye group,the masculine rate of CCR5 was negatively correlated with BUT and S Ⅰ t (r=-0.473,-0.385,both at P＜0.05).The masculine rate of CXCR3 was negatively correlated with BUT and S Ⅰ t (r =-0.753,-0.684,both at P＜0.05).No considerable correlations between the positive rate of CCR5 with the positive rate of CXCR3 or FLS scores (r =0.231,0.336,both at P＜0.05).The relative expression levels of RANTES,MIP-1α,MIG and IP10 mRNA in the dry eye group were significantly higher than those in the normal control group (t =3.091,2.894,2.688,2.245,all at P＜0.05),but there were no significant differences in the relative expression levels of MIP-1β and I-TAC mRNA between the two groups (t =0.512,1.979,both at P＞0.05).The positive correlations were seen between the masculine rate of CCR5 with the relative expression of RANTES or MIP-1α mRNA (r=0.473,0.285,both at P＜0.05),but there was no obvious correlation between the masculine rate of CCR5 and the MIP-1 β mRNA expression(r=0.214,P＞0.05).In addition,the masculine rate of CXCR3 was positive correlated with the expressions of MIG and IP10 mRNA (r=0.553,0.314,both at P＜0.05),whereas the masculine rate of CXCR3 was not related to the expression of I-TAC mRNA (r=0.364,P＞0.05).Conclusions Dry eye is probably along with the long-term infiltration of inflammatory cells.The delayed allergy induced by Th1 cells and the nature killed cells is probably the primary cause to xerophthalmia.CCR5,CXCR3 and their ligands might be the regulative targets in the inflammation mechanism of dry eye.

Objective To explore the change of serum level of neuron specific enolase (NSE) in neonates with asphyxia before and after head mild hypothermia.Methods Eighty-two asphyxial neonates were selected,including 39 mild asphyxial neonates and 43 severe asphy-xial neonates,and 29 healthy neonates were selected as control group.Forty-three severe asphyxial neonates were randomly assigned into mild hypothermia treatment group and traditional treatment group.Neonates in traditional treatment group were just given traditional treatment.While neonates in mild hypothermia treatment group received head mild hypothermia therapy and their nasopharyngeal temperature were maintained at(34.0 ? 0.5) ℃ for 72 h.Before treatment and 72 h after treatment,2 mL blood was collected,and the serum NSE was determined by radio immunoassay.Results NSE levels in mild asphyxial neonates group[(34.83?6.17) ?g/L] and severe asphyxial group[(59.58?8.87) ?g/L] were significantly higher than that of control group[(30.57?4.88) ?g/L](t=3.07 P0.05).The level of NSE at 72 h in severe asphyxial neonates with head mild hypothermia therapy[(40.97?6.55) ?g/L] was significantly lower than that of traditional treatment group [(48.15?5.57) ?g/L](t=3.86 P

Objective To investigate the protective effect of Schizandrae Lignanoid (SCL) on myocardial ischemia reperfusion injury in hyperlipidemic rats and its mechanisms and provide theoretical basis and experiment foundation for treatment of myocardial ischemia complicated with hyperlipidemic disease with SCL in the future. Methods After hyperlipidemic rat models were set up by oral administration of high lipid emulsion,the rats were divided into seven groups: control group,sham group,model group,SCL 60,20,5 mg?kg-1 groups and 200 mg?kg-1 CDT group.After reperfusion,the blood samples taken from the ventricles were assayed for blood lipid;MPO activities of ischemic myocardium were measured;Infarct area of left ventricles was measured by Evens blue-TTC staining,and myocardial pathological changes were also observed.Results Compared with model group, the myocardial infarct area/ventricle area ratios and MPO activities in SCL 60,20,5 mg?kg-1 groups decreased (P

Background Age-related macular degeneration (AMD) is a group of vision-threatening eye disorders.Previous researches showed that the activation of Akt/mammalian target of rapamycin (Akt/mTOR) pathway closely related to the mechanism of AMD.A new specific method to inhibit Akt/mTOR pathway will become a breakthrough for the treatment of AMD.Objective This study was to establish a mouse fibroblast cell line (NIH/3T3) which can stably inhibit the expression of mTOR gene and provide a cell model for the study on the function of Akt/mTOR pathway in AMD and observe the influence of mTOR gene knockdown on related proteins.Methods Three short hairpin RNAs (shRNAs) targeting mTOR gene were designed and synthesized based on murine mTOR mRNA sequence.Double-strand shRNA hairpins were separately cloned into PIGZ-green fluorescent protein (GFP) +Puro vectors to produce recombination plasmids.The packaged lentiviral plasmids and RNAi plasmids were co-transfected into NIH/3T3 cells,a mouse fibroblast line.After puromycin selection and culture expansion,0.5 mg/L puromycin was added the culture medium to establish stable cell clones.The expressions of mTOR mRNA and protein in NIH/3T3 cells were detected by real-time PCR and Western blot respectively,and the inhibitory efficiency of interference was analyzed.Results Transfected GFP-labeled NIH/3T3 cells by lentiviral presented the green fluoresccence with the efficiency of infection of 90％ in the third day.Real-time PCR showed a distinct band of mTOR mRNA in 184 bp.The knockdown rate for sh1,sh2 and sh3 were respectively 31.3％,31.8％ and 45.3％ in the lower multipolicity of infection (MOI) group ;while in the higher MOI group,the knockdown rate for sh1,sh2 and sh3 were 47.1％,56.5％ and 71.6％ respectively.Western blot assay exhibited weakened expression band of mTOR protein in NIH/3T3 cell line for sh1,sh2 and sh3 in both lower and higher MOI groups with the weakest expression for sh3.Conclusions A stable mouse fibroblast cell line is established by the inhibition of mTOR with RNA interference technique,which can provide a cell model for studying the function of Akt/mTOR pathway in AMD.

Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.
Absorption ; Astragalus Plant ; chemistry ; Biological Transport ; Caco-2 Cells ; Fluorescein-5-isothiocyanate ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Plant Extracts ; chemistry ; pharmacokinetics ; Polysaccharides ; chemistry ; pharmacokinetics

Objective To investigate the clinical characteristics and mutations of guanosine triphosphate eyclohydrolase (GCH) Ⅰ gene in patients with dopa-responsive dystonia (DRD). Methods Five families with 18 affected family members and 17 patients with sporadic DRD were examined. Patients were allocated into 3 groups according to onset time, either in childhood, or in adolescence or adult. Interview, physical examination, psychologic testings and CT/MR scan were performed. Mutation screening was performed on 26 patients and 1 normal family nember. Thirty-five healthy control subjects were matched for age and sex. Statistical analysis were conducted with the use of SPSS 13.0 computer software. Results(1)Most of patients started with dystonia. The main clinical manifestation was dystonia too. There was no difference among 3 groups.(2) There were significant differences in diurnal fluctuation among 3 groups(15/15,6/6,7/14, χ2=13.125,P=0.001). Diurnal fluctuation negatively correlated with age (r=-0.720, P<0.01).(3)The differences in postural tremor were also found among 3 groups (7/15,5/6,1/14, χ2=8.073, P=0.018). Postural tremor positively correlated with age (r=0.399, P=0.018).(4)There were differences in exaggeration of tendon among three groups(11/15,1/6,4/14, χ2=8.309, P=0.016). Exaggeration of tendon reflexes negatively correlated with age (r=-0.429, P=0.010).(5)The scores of Hamilton Depression Scale and Hamilton Anxiety Scale in patients were higher than those in controls.(6)DNA sequencing revealed a heterozygous A224G missense mutation(Tyr75Cys)located within exon 1 in one autosomal dominant inheritance family. Conclusions The manifestations of DRD varies. The clinical course is closely correlated with age. A missense mutation(A224G)in coding region of the GCH 1 gene probablyleads to the occurrence of DRD.

The effective components are the material basis for efficacy of Traditional Chinese Medicine (TCM), also the core of its quality control. This paper focused on two key scientific issues that how do the effective components form?, How can we improve the effective components?, combining with examples of our group's research, to dis-cuss the research thoughts, technical methods for TCM quality-design. This paper provided a methodological refer-ence for the study of TCM quality-control.