Objective To investigate the anti-H5N1 activities and chemical constituents from stems and leaves of Buddleja lindleyana Fort. Methods Constituents were separated through AB-8 macroporous resin, chromatography of silica gel, Sephadex LH-20 and recrystallization. Structures of the compounds were identified by analysis of physicochemical properties and spectral data. Results Ten compounds were isolated and identified as linarin(l), rutin(2), luteolin(3), quercetin(4), apigenin(5), hesperetin (6), salidroside (7), oleanolic acid (8), ß-sitosterol (9), and daucosterol (l0), respectively. Conclusion Compounds 2-7 were obtained from this plant for the first time.

Objective: To establish an extraction process optimization method based on fingerprint combined with principal component analysis, which was finally applied to Compound Huzhang ethanol extraction process optimization. Methods: Taking Compound Huzhang prescription as model drug, different ethanol extraction condition fingerprint was established by HPLC, harvesting the areas of the common peaks, the total factor scores were calculated by PCA. Arranged experiments with U9(93×31) uniform design method, choosing the total factor scores as index, the influence of extration times (X1), alcohol consumption(X2), alcohol concentration(X3), and time of extracting(X4) on the yield of extract was investigated, then the technological parameters of optimum ethanol extraction condition were selected by multi-nonlinear mathematic models. Results: Multi-nonlinear mathematic models described the relationship between response indexes and factor variables with a regression coefficient of 0.997. The optimized conditions for Compound Huzhang Prescription were extracted in 90% ethanol six times as much as it for three times, 1h once. Conclusion: It is proved that the extraction method is suitable and feasible that could provide a reference for compound Chinese medicine extraction process optimization.

Objective: To observe the feasibility of acellular cartilage extracellular matrix (ACECM) oriented scaffold combined with chondrocytes to construct tissue engineered cartilage.

Objective: To analyze the dynamic changes of inflammatory factors in rats model induced with IBS-D. Methods: Thirty Wistar rats were randomly divided into the blank group, the IBS-D group, and the TAK-242 group. All rats received acute and chronic stress method, followed by no further treatment for the blank group. To induce IBS-D, before performing acute and chronic stress, the IBS-D group received 3 mg/kg saline. To explore the potential mechanism of TLR4 in IBS-D, TAK-242, an antagonist of TLR4 was given to 3 mg/kg TAK-242 group after they received acute and chronic stress. Fecal traits were evaluated by Bristol classification at 0, 7, 14, 21, 28 days, and the levels of MyD88, IL-1β, IL-6 were quantified by Elisa assays; and the levels of TLR4, NF-κB were detected by Western blot. Results: In the blank group, there was no significant change in the scores of Bristol stool and expression levels of MyD88, IL-1β, IL-6, TLR4 and NF-κB at the 7th, 14th, 21st and 28th day. Compared with the blank group, in the IBS-D rats and TAK-242 rats, the scores of Bristolian stools (P<0.05), the levels of MyD88 and serum inflammatory factors (P<0.05), TLR4, NF-κB were all increased (P<0.05). However, the scores of Bristolian stools and the expression levels of MyD88, IL-1β, IL-6, TLR4, and NF-κB in the TAK-242 group were lower than those in the IBS-D group, suggesting therapeutic effects of TAK-242 in IBS-D. Conclusions: IBS-D may increase inflammatory factors through activating the TLR4/MyD88/NF-κB signaling pathway, resulting in disease progression.

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) and "Feishu" (BL 13) on pulmonary function, inflammatory reaction and expression of macrophage migration inhibitory factor (MIF) and its receptor complex CD 74-CD 44, etc. in rats with chronic obstructive pulmonary disease (COPD), so as to explore its mechanism underlying improvement of COPD. METHODS: Thirty male SD rats were randomly divided into normal, model and EA groups (n＝10 in each group). The COPD model was established by intratracheal infusion of Lipopolysaccharide (LPS, 1 mg/mL) and forced smoke-inhaling. EA was applied to bilateral ST 36 and BL 13 for 30 min, once daily for 7 days. The rat's lung function (forced vital capacity [FVC], forced expiratory capacity ratio ([FEV 0.1/FVC] and [FEV 0.3/FVC]) was detected under anesthesia. Pathological changes of the lung tissue were detected by H．E. staining, and the contents of MIF, tumor necrosis factor-α (TNF-α), interleukin-1 β (IL-1 β) and IL-8 in serum, bronchoalveolar lavage fluid (BALF) and lung tissue were assayed by ELISA. The immunoactivity of CD 74 and CD 44 was detected by immunohistochemistry, and the expression levels of MIF, CD 74, CD 44 and p 38 MAPK mRNAs and proteins were examined by quantitative RT-PCR and Western blot, respectively. RESULTS: Compared with the normal group, the FVC, FEV 0.1, FEV 0.3, FEV 0.1/FVC and FEV 0.3/FVC levels were significantly decreased in the model group (P<0.01). After EA treatment, the FVC, FEV 0.1, FEV 0.3, FEV 0.1/FVC and FEV 0.3/FVC were significantly increased (P<0.01, P<0.05), suggesting an improvement of the pulmonary function after EA. H．E. staining showed that the severity of modeling induced alveolar expansion and inflammatory cell infiltration in the lung tissue was relatively milder in the EA group relevant to the model group. The contents of MIF, TNF-α， IL-1 β and IL-8 in the serum, BALF and lung tissues were significantly higher in the model group than in the normal group (P<0.01), and significantly down-regulated in the EA group relevant to the model group (P<0.01). The expression levels of MIF, CD 74, CD 44 and p 38 MAPK mRNAs and proteins and the immunoactivity levels of CD 74, CD 44 in the lung tissue were obviously higher in the model group than those in the normal group (P<0.01), and considerably lower in the EA group than those in the model group (P<0.01). There was a positive correlation between p 38 MAPK and MIF in mRNA and protein expression levels (P<0.01). CONCLUSION: EA intervention can improve the pulmonary function in COPD rats, which may be related to its effects in inhibiting inflammatory reaction, and MIF/CD 74-CD 44/p 38 MAPK signaling pathway.

Objective: To investigate the impact of electrical stimulation numbness to central nervous system in normal people. Methods: Electric stimulator was used to establish low-frequency, high-frequency numbness models with 22 healthy subjects. Scores for numbness and emotional valence for different electrical stimulations were recorded, and data of functional MRI (fMRI) in task state were also collected. The differences of numbness and emotional valence score of subjects under low frequency and high frequency electrical stimulation were compared, respectively. Then brain regions with significant differences in brain activation intensity under different frequencies of electrical stimulation were obtained, and the differences of activation effect values of activated brain regions under different electrical stimulations intensity were compared, respectively. The relationship of numbness and emotional valence scores under high frequency electrical stimulation were analyzed. Results: There were statistical differences of numbness (t=13.18) and valence score (t=10.77) under different electrical stimulations (both P<0.05). There was negative correlation between numbness and valence scores under high-frequency electrical stimulation (r=-0.53, P=0.01). The brain areas with significant differences of activation intensity under different electrical stimulation included left parietal operculum, left middle cingulate gyrus, left temporal gyrus, right parietal operculum, left postcentral gyrus, right central operculum, right posterior insula and left thalamus (all P<0.05). Conclusion: The activated brain regions under different electrical stimulations caused numbness include left parietal operculum, left middle cingulate gyrus, left temporal gyrus, right parietal operculum, left postcentral gyrus, right central operculum, right posterior insula and left thalamus in normal people. Block-designed BOLD-fMRI can be used to detect activity of brain areas associated with numbness.

OBJECTIVE: To provide reference for patients with type I incisions about rational use of antibiotics.METHODS: Between Oct.2007 and Mar.2008,a total of 1 024 medical records of patients with type I incisions from orthopaedics,common surgery,department of gynecology in 12 hospitals of Xining area were sampled for analysis of prophylactic use of antibiotics.RESULTS: 100.0% of the type I incision patients received prophylactic antibiotics;28.4% used antibiotics without indication;16.9% used antibiotics at 0.5～2 h before operation;33.4% used antibiotics at more than 2 h before operation;and 49.7% received antibiotics postoperatively rather than preoperatively.The irrational and nonstandard use of antibiotics manifested as improper in the choice of drug variety,nonstandard in drug combination,irrational in dosage and administration,prolonged use of antibiotics,lacking basis for the change of drug variety etc.CONCLUSION: The prophylactic use of antibiotics in patients with type I incisions from 12 hospitals of Xining area is far from perfect,therefore,it is urgent to strengthen the standard management on the use of antibiotics.

Aim To investigate the effects of DNMT3A regulating Drp1 mediated mitochondrial fission on the proliferation and migration of active hepatic stellate cells. Methods HSC-T6 cells were activated by 5 μg·L-1 TGF-β1 for 24 h, and DNMT3A lentivirus infection model was established to silence DNMT3A. The experiment was divided into control group, TGF-β1 group, TGF-β1+LV5-NC group and TGF-β1+ LV5-DNMT3A group. The effects of DNMT3A on related mRNA and protein expression were detected by RT-qPCR and Western blot. The cell proliferation was detected by CCK-8. The effect of DNMT3A on the migration ability of HSCs cells was observed by Wound healing assay and Transwell migration assay. Results Lentivirus infection successfully constructed a DNMT3A silencing model. Compared with the control group, the level of DNMT3A significantly increased, the mRNA and protein levels of the fibrosis markers collagen and α-SMA in the TGF-β1 group significantly increased, and the mRNA and protein levels of the mitochondrial fission marker Drp1 significantly increased; At the same time, the proliferation and migration ability of HSCs cells was significantly improved. Compared with the NC group, the DNMT3A level of the DNMT3A silenced group was significantly reduced, the expressions of collagen I, α-SMA and Drp1 were significantly inhibited, and the proliferation and migration capabilities of HSCs were also significantly inhibited. Conclusions Silencing DNMT3A inhibits the level of Drp1 and inhibits the proliferation and migration of HSCs at the same time. It is suggested suggest that DNMT3A-mediated low level DNA methylation modification may inhibit the occurrence of mitochondrial fission by inhibiting the level of Drp1, thereby inhibiting the activation of HSCs and affecting the occurrence and development of liver fibrosis. ,,,,.

OBJECTIVETo explore the feasibility of amplifying the leukemia tumor associated antigens-specific cytotoxic T lymphocytes (TAA-CTL) ex vivo and to evaluate the cytotoxicity of TAA-CTL.

METHODSThe peripheral blood mononuclear cells were enriched by density gradient centrifugtion; TAA-CTL were generated by stimulation of PBMNC with peptide-pulsed DC at an effector-to-target ratio of 10:1; immunophenotype of TAA-CTL was analyzed by flow cytometry; cytotoxicity assay was used to evaluate the cytotoxic activity of TAA-CTL against peptide-pulsed autologous target cells (PHA-Blasts).

RESULTSTAA-CTL expanded from volunteer showed a mean expansion of 3.81±1.61, the phenotyping of the TAA-CTL was predominantly CD3+ (97.22±0.71)% with varying content of CD4+ (41.47±27.08)% and CD8+ (56.40±11.15)% T cells, it also contained few nature killer cells (0.50±0.31)% and rare residual B cells (0.14±0.20)%; the subpopulations of TAA-CTL and CTL were not statisticaly significantly different in the proportion (P>0.05); the detection of intracellular cytokines after stimulation with peptide showed that the secretion rates of IFN-γ and TNF-α in CD8+ TAA-CTL were (27.67±2.21)% and (34.2±0.71)%, while the secretion rates were (21.6±2.55)% and (9.97±3.44)% in CD4+ TAA-CTL. Compared with the CD8+ TAA-CTL group, the secretion rates of IFN-γ and TNF-α were (1.36±0.04)% and (5.58±0.03)% in CD8+ CTL, the rates of IFN-γ and TNF-α were (0.91±0.06)% and (1.60±0.07)% in CD4+ CTL. The secretions of IFN-γ and TNF-α in CTL were both significantly lower than those in TAA-CTL (P<0.01); the specific killing efficiency of the TAA-CTL against TAA-pulesd target cells were (77.00±1.00)%, (67.40±3.60)%, (60.55±2.45)% and (26.85±5.25)%, when the effecto-target ratios were 40:1, 20:1, 10:1 and 5:1, and there was negligible lysis of TAA-CTL for PHA-blast (P<0.01).

CONCLUSIONTAA-CTL can be successfully induced and generated ex vivo from the healthy volunteer peripheral blood, and the TAA-CTL possess a specific killing activity.

Objective : To evaluate the anti⁃inflammatory effect of tissue inhibitor of metalloprotease (TIMP1) on a rat model of irreversible pulpitis. Methods : An irreversible pulpitis model was established by pulp exposure on maxillary first molars of 8 ⁃10 weeks old male rats , and pulpotomy was performed at 24 h , 36 h and 48 h after pulp exposure , with TIMP⁃1 covering the pulp section (TIMP⁃1 group) and saline as control (NS group) . Coronal obturation were performed with glass⁃ionomer cement. Histological and immunohistochemical analysis were performed at 24 and 48 h after administration. Results : Quantitative histopathological analysis of HE⁃stained sections showed that the inflammation score (inflammatory cellular response of the root pulp , morphological changes in root pulp tissue and extent of root pulp necrosis) of the pulp were higher in the TIMP⁃1 group than those in the NS group (P < 0. 001 or P < 0. 01) . Immunofluorescence staining (CD68⁃labeled) sections showed that the intensity of CD68 immunofluorescence was higher in the TIMP⁃1 group than that in the NS group (P < 0. 001) . Conclusion TIMP⁃1 had no anti⁃inflammatory effect in a rat model of irreducible pulpitis and increased the degree of inflammation in the root pulp.