Objective To investigate the effects of pretreatment with different concentrations of sufentanil of ketamine on the differentiation of human helper T cells in vitro. Methods Twenty-two healthy volunteers (11 males, 11 females) aged 20-45 yrs were enrolled in this study. In each volunteer 20 ml of blood was taken from peripheral vein and divided into 7 groups: control group (0.9% NaCl), 3 sufentanil groups (0.05, 0.5, 5.0 ng?ml-1) and 3 ketamine groups (100, 500, 2 500 ng?ml-1) .Whole blood and mononuclear cells from peripheral blood (PBMCs) were incubated in the presence of 0.9% NaCl or different concentrations of sufentanil or ketamine for 24 h. Then the stimulants-phorbolmyristate + lonomycin + glgistap (inhibitor of intracellular protein transport) were added to whole blood and phytohemagglutinin (PHA) was add to PBMCs. The whole blood was incubated for another 4h and PBMCs were incubated for another 48 h. Then the T-lymphocytes were collected for determination of intracellular level of IFN-?(as a marker of Th1 cells) and IL-4 (as a marker of Th2 cells) in the whole blood using three-color flow cytometry and the expression of CCR5 + (as a marker of Th1 cells) and CCR3 + (as a marker of Th2 cells) in PBMCs. The Th1/Th2 ratio was calculated. Results In sufentanil 0.5 and 5.0 ng?ml-1 groups the percentage of Th2 cells was significantly increased while the percentage of Th1 cells and Th1/Th2 ratio were significantly decreased. In ketamine 500 and 2 500 ng?ml-1 groups the percentage of both Th1 and Th2 cells were significantly decreased and the Th1/Th2 ratio was significantly increased. Conclusion Sufentanil can encourage helper cells to differentiate into Th2 cells while ketamine inhibit the helper cells to differentiate into Th1 and Th2 cells, especially the Th2 cells in a dose-dependent manner.

Objective: To explore early diagnostic value of combined detection of ischemia-modified albumin and other biochemical markers for acute coronary syndrome (ACS). Methods: A total of 156 patients, who received treatment within 4h because of chest pain onset, were divided into ACS group (n=112) and non-ischemic chest pain group (NICP group, n=44) according to coronary angiography (CAG). Blood sample was taken within 4h and during 4~8h after chest pain to measure cardiac biochemical markers, including ischemia-modified albumin (IMA), LDH, CK, CK-MB and cardiac troponin I (cTnI), comprehensive analysis was performed by detected results of above biochemical markers and CAG. Results: For diagnosis of ACS, within 4h after onset IMA possessed highest sensitivity (87.50%) and highest accuracy (80.77%), cTnI possessed highest specificity (95.45%); In 4～8h IMA still possessed highest sensitivity (91.07%), cTnI still possessed highest specificity（97.73%）, CK（85.90%），and IMA（85.25%）possessed highest accuracy （both CK and IMA had no significant difference）; According to comprehensive analysis, regardless of within 4h or during 4~8h after chest pain, IMA possessed highest sensitivity and highest accuracy, cTnI possessed highest specificity. Within 4h after chest pain, detection of IMA combined above-mentioned other four indexes increased sensitivity (89.28%), specificity （95.45%） and accuracy （91.02%）to highest level. Conclusion: For diagnosis of ACS, IMA possesses highest sensitivity and highest accuracy, cTnI possesses highest specificity; IMA combined above-mentioned other four indexes may increase sensitivity, specificity and accuracy to highest level.

In order to reveal the treatment mechanism of Chinese medicine with the effect of activating blood and resolving putridity, we selected acetyl-11-keto-beta-boswellic acid (AKBA) and arsenic trioxide (ATO), the main monomeric components of frankincense and arsenolite which are two most commonly used Chinese medicine with effect of activating blood and resolving putridity. We combined AKBA and ATO as a compound, and explored its regulatory role in productions and activities of matrix metalloproteinase (MMP)-1, MMP-2 and MMP-9 in human skin fibroblasts (HSFbs) and human acute monocytic leukemia cell line THP-1 in inflammatory state.

The Fe-N-C composite catalyst was prepared by the thermal decomposition of the chelate precursors based on Fe central ions and o-phenylenediamine ligands.It was observed from the scanning electron microscopy that the crumpled carbon micro-and nano-sheets were intertwined to form a free-standing tremella-like 3D structure.The N2 adsorption/desorption experiments revealed that the composite contained ample micro-and meso-pores and had a specific surface area of 290 m2/g.Graphitic C and multi-crystal Fe3C as main components were confirmed by the X-ray diffraction, and N-doping in the general form of graphite N and pyridine N was also verified by X-ray photoelectron spectroscopy.The electrochemical measurement showed that the tremella-like Fe-N-C composite catalyzed oxygen reduction through a four-electron path in an alkaline solution, and its activity was comparable to the commercial Pt/C catalyst.After 2000 cycles, the limited current density of the Fe-N-C catalytic electrode only decreased less than 5%, and the half-wave potential shift negatively 5 mV, which suggested that the Fe-N-C composite catalyst had better catalytic stability than the commercially used Pt/C catalyst.

Objective To investigate the application of venereal disease research laboratory(VDRL) test and toluidine red unheated serum test(TRUST) in syphilis laboratory diagnosis.Methods Serum,plasma and cerebrospinal fluid (CSF) in syphilis patients and healthy controls were measured by VDRL and TRUST.Results The VDRL detection results in serum and CSF sepcimens were generally higher than the TRUST detection results by 1-2 titers,while in plasma specimen,the VDRL detection results were generally lower than the TRUST detection results by 1-2 titers than TRUST when using plasma specimen.In addition,the VDRL detection in normal control serum and plasma specimens all appeared different degrees of false positive,but in the detection of normal control CSF,the results of TRUST and VDRL were consistent.Conclusion TRUST is more suitable for serum and plasma specimens,and CSF is suitable for the CSF specimen,but not suitable for serum and plasma detection.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, which as a group can degrade essentially all extracellular matrix components. The proteolytic property of the MMPs is important during wound healing to remove debris and facilitate cell migration. Targeting towards the decreased MMPs activities is a new treatment strategy for healing chronic wounds. Salvia miltiorrhiza is a popular Chinese herb that could promote chronic ulcers healing for topical use. Salvianolic acid B (Sal B) is the most abundant bioactive component in Salvia miltiorrhiza. The research was designed to explore the inhibitory effects of Sal B on MMP-1, MMP-2 and MMP-9 activities.

AIM: To evaluate the effects of acetyl-11-keto-beta-boswellic acid (AKBA, a main active component from frankincense, one of the traditional Chinese herb for healing wounds) on the activities of matrix metalloproteinase(MMP)-1, MMP-2 and MMP-9.METHODS: Pure human interstitial collagenase (MMP-1) or gelatinase A (MMP-2) was activated by p-aminophenylmercuric acetate (APMA), and was incubated with AKBA for 1 h. The activities of the enzymes were observed by quenched fluorescent substrate. The lysates of rat polymorphonuclear neutrophils [PMNs, rich in gelatinase B (MMP-9)] was incubated with AKBA for 1 h, and activity of MMP-9 was tested by gelatin zymography. Three cell models: activated human dermal fibroblasts by TNF-α, activated THP-1 cells by PMA and fibroblasts-THP-1 co-culture system were established. AKBA was cultured with these cell models for 24 h. The levels of MMP-1, MMP-2 and MMP-9 in the cell culture supernatants were tested by ELISA and activities of MMP-2 and MMP-9 were tested by gelatin zymography assays.RESULTS: AKBA dose-dependently inhibited the activities of human MMP-1 and MMP-2 at the range of 0.1-0.8 mmol/L, with 50% inhibitory concentration (IC50) of 0.18 mmol /L and 0.27 mmol/L, respectively. In the range of 0.05-0.85 mmol/L, AKBA inhibited the MMP-9 activity (P<0.01). Although AKBA promoted fibroblasts to secrete MMP-2, the production of MMP-9 by THP-1 was inhibited. In the cell co-culture system, the inhibitory effects on MMP-1, MMP-2 and MMP-9 productions were also observed.CONCLUSION: AKBA, as a bioactive component of frankincense, has an inhibitory effect on MMPs production and activities, indicating the possible mechanism for healing chronic wounds by frankincense.

Objective To explore the role of MKK34 (a peptide spanning a C-terminal α-helical region in TSLP) on airway inflammation and β-catenin of airway epithelium in a HDM-induced mouse asthma.Methods 32 male BALB/c mice were randomly divided into control,MKK34,asthma and MKK34 + HDM groups.The mice in the asthma group were exposed to HDM for five consecutive days and the MKK34 + HDM group was pretreated with MKK34 1 h prior to the HDM intranasally treated.After 8 weeks' treatment,animal lung function test and pathological staining were performed to evaluate the asthma situation,IL-4,IFN-γin bronchoalveolar lavage fluid and IgE in the serum were detected,immunohistochemistry and western blot were used to assess β-catenin and p-ERK,t-ERK levels.Results Airway reactivity,IL-4 and IgE in the asthma group were significantly higher than that in the control group.Treatment with MKK34 significantly decreased airway hyperresponsiveness,IL-4 and IgE.HE staining demonstrated the chronic bronchitic inflammation in the lungs of asthma group.β-catenin in the control group was distributed evenly at the cytomembrane of epithelial cells.In the asthma group,β-catenin was disordered in epithelial cells and its expression was decreased.Treatment with MKK34 ameliorated the damage of β-catenin and chronic bronchitic inflammation.The protein levels of p-ERK1/2 increased obviously in the asthma group.The pretreated group significantly decreased the expression of p-ERK1/2.Conclusions MKK34 can ameliorate the airway inflammation and the destruction of β-catenin of airway epithelium in a HDM-induced mouse asthma.The ERK pathway may play a role in this process.

Objective To investigate the impact of 1,25(OH)2D3 on histological changes and activation of STAT3 in BLM?induced pulmonary fibrosis mice. Methods 30 male C57BL/6 mice were randomly divided into control group ,BLM group and BLM+VD group. Mice in BLM group and BLM+VD group received intratracheal injection of BLM(3 U/kg). Control group were intratracheally injected equal volume of sterile saline. From the first day after the surgery,mice in BLM+VD group received intraperitoneal injection of VD (5μg/kg·d). After 21 days, H&E and Masson′s trichrome staining were carried out. Aschroft score were used to evaluate histological changes in lungs. IL?6,IL?4 and INF?γin BALF were assessed by Elisa. p?STAT3,α?SMA and Collagen I were detected by western blot (WB) and immunohistochemistry. Results Fibrosis score and level of α?SMA,Collagen I in BLM group were significantly higher than that in control group (P < 0.05). However ,treatment with VD effectively at?tenuated fibrosis (P<0.05). IL?6 and IL?4 increased while INF?γwas decreased in BALF of BLM group (P<0.05). VD could ameliorate these changes. Upregulation and neuclear translocation of p?STAT3 were observed in BLM group,while VD intervention could inhibit phosphorylation of STAT3. Conclusions VD attenuate BLM?induced pulmonary fibrosis and regulate inflammatory cytokines probably by blocking STAT3 activation.

The aim of the present study was to evaluate the effect of storage conditions,pretreatment,temperature,time and excystation solutions on in vitro excystation of Cryptosporidium oocyst.Cryptosporidium andersoni oocyst was used as a model and the results showed that 0.5% sodium hypochlorite could enhance the excystation rates.But there was no significant difference compared with oocysts untreated by sodium hypochlorite(P＞0.05).0.75% synthetic sodium taurocholate and 1% bile solution could urge the excystation of oocysts,which were significantly different compared with the excystation rate of oocysts in 0.25% trypsin solution or in PBS(P<0.05).The excystation rates of oocysts in acidic water (pH =3) were similar with the rates in PBS (pH =7.2) but significantly different from the rates in alkaline water (pH =9) (P<0.01).Additionally,the excystation rate of oocysts in water of 24℃ was significantly lower than in water of 37℃(P<0.01),and the excystation rate of oocysts raised gradually at 37℃ with the passage of time.It's concluded that temperature,acidity and excystation solution were vital factors for the in vitro excystation of Cryptosporidium oocyst.A higher excystation rate could be observed when oocysts were pretreated with 0.5% sodium hypochlorite and treated with 0.75% synthetic sodium taurocholate at 37℃ for 3 hours.