AIM: To observe the role of c-fos in the protection by cholecystokinin-octapeptide (CCK-8) against pulmonary artery smooth muscle cell (PASMC) injury induced by lipopolysaccharide (LPS). METHODS: The ultrastructure of PASMC was observed under transmission electron microscope (TEM). MDA content,LDH release and the rate of trypan blue in PASMC were measured,and immunocytochemistry technique was adopted to observe the c-fos protein expression. RESULTS: The TEM results showed significant PASMC structural injury in LPS group and alleviated structural changes in LPS+CCK-8 group. CCK-8 reduced the increase in the rate of trypan blue uptake,MDA content and LDH release in PASMC induced by LPS. LPS lightly increased c-fos protein expression,which was enhanced by CCK-8. CONCLUSION: CCK-8 attenuated the injury of PASMC induced by LPS,which may be concerned with the increase in c-fos protein expression.

Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.

AlM: To monitor long - term changes of matrix metalloproteinase-2 ( MMP-2 ) in human tears fluid after laser in situ keratomileusis ( LASlK) .
METHODS: Thirty - two myopia cases ( 64 eyes ) underwent uneventful LASlK were enrolled in the study. Tear fluid were collected and MMP-2 expression was analyzed by Western - bolt assay preoperatively and postoperatively on 15d, at 1, 3mo, and 1a.
RESULTS:LASlK increased the concentration of MMP-2 in human tear fluid. At 15d postoperatively, the magnitude of MMP-2 was 1. 4 times that of preoperative, thereafter subsided, but didn't return to preoperative level by 3mo ( P < 0. 05 ). Up to 1a after surgery, the concentration of MMP-2 almost recovered (P>0. 05).
CONCLUSlON: MMP- 2 is significantly expressed in human tear fluid after LASlK, then subsided with time, but didn't return to preoperative level by 3mo and almost recovered up to 1a, indicating wound healing of LASlK would continue up at least 3mo after surgery and almost recovered 1a postoperatively.

To study enhancing effect of IL-3 gene transfected bone marrow stromal cell which can be induced by doxycycline (Dox) to express IL-3 cytokine on the proliferation and differentiation of hematopoietic stem cell, retrovirus vector system contained mIL-3 cDNA was established and bone marrow stromal cell line was transfected, and obtained QXMSC1Tet-on-IL-3, in which expression level of IL-3 can be modulated by Dox. The activities of IL-3 were measured under different Dox concentrations. The numbers of hematopoietic progenitors (CFU-GM, CFU-E, CFU-GEMM and LTC-IC) were measured and the capacity of QXMSC1Tet-on-IL-3 sustaining hematopoietic progenitor cell growth was evaluated. The results showed that IL-3 gene transfected stromal cell line QXMSC1-Tet-on + IL-3 expressed high concentration of IL-3 in vitro under control of Dox. The supernatant of QXMSC1-Tet-on + IL-3 was able to increase the number of CFU-GM, CFU-E and CFU-GEMM. The total numbers of nucleated cells and long-term cultured colonies increased in LTC-IC assay. It is concluded that in the culture of QXMSC1-Tet-on + IL-3 cells, Dox actually enhanced IL-3 expression, and thus augmented the proliferation and differentiation of hematopoietic stem/progenitor cells in vitro.
Animals ; Bone Marrow Cells ; physiology ; Cell Differentiation ; drug effects ; Doxycycline ; pharmacology ; Hematopoiesis ; drug effects ; Hematopoietic Stem Cells ; cytology ; Interleukin-3 ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Stromal Cells ; Transfection

Recent data have revealed that inhibiting autophagy exacerbates lipid accumulation in hepatocytes and nitrite treatment reduces total triglyceride levels in the high-fat diet mice. Therefore, the present study aimed to determine the effects of nitrite on simple hepatic steatosis and the possible role of autophagy. Firstly, steatotic L-02 cells were induced by incubating L-02 cells with 1.2 mmol · L(-1) oleic acid (OA) for 24 h. Secondly, steatotic L-02 cells were treated with 0.2 mmol · L(-1) sodium nitrite (SN) plus 3-methyladenine (3-MA), or chloroquine (CQ) for 24 h, and then lipid accumulation was measured with oil red O staining and triglyceride quantification. The notable steatosis could be observed in L-02 cells following exposure to 1.2 mmol · L(-1) OA for 24 h. Treatment with 0.2 mmol · L(-1) sodium nitrite reduced lipid accumulation in steatotic L-02 cells. 3-MA weakened the ability of sodium nitrite to ameliorate hepatic steatosis. Additionally, the sodium nitrite increased number of LC3-II immunostaining puncta and LC3-II protein expression was confirmed by immunofluorescence or Western blot analysis, and the effects were enhanced by CQ treatment. The number of increased cytoplasm vacuoles and lysosomes increased was confirmed by phase contrast and fluorescence microscope respectively. The increased autolysosome was detected by electron microscopy, this phenomenon could be reversed by CQ treatment. These data demonstrated that sodium nitrite enhanced the autophagic flux and decomposition of triglycerides in steatotic L-02 cells.
Adenine ; analogs & derivatives ; Autophagy ; Blotting, Western ; Cells, Cultured ; Chloroquine ; Cytoplasm ; Fatty Liver ; Hepatocytes ; drug effects ; Humans ; Lipid Metabolism ; drug effects ; Microscopy, Fluorescence ; Microtubule-Associated Proteins ; metabolism ; Oleic Acid ; Sodium Nitrite ; pharmacology ; Triglycerides

AIMTo establish a sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of isomeric impurity in raw drug material of cefepime dihydrochloride.

METHODSThe HPLC separation experiments were performed on a reversed phase C18 column, with acetonitrile-10 mmol x L(-1) ammonium acetate (5:95) as mobile phase (a flow rate of 0.8 mL x min(-1)). The analytes were determined by electrospray ionization tandem mass spectrometry in positive mode. The chromatogram and mass spectra of cefepime dihydrocloride and its isomeric impurity were obtained by LC-MS/MS.

RESULTSThe method can be used for the separation and identification of cefepime dihychrocloride and its isomeric impurity, of which the retention times were 15.28 min and 9.18 min, respectively. Based on the MS/MS spectra of their molecular ions, the different fragmentation pathways of cefepime dihydrocloride and its isomeric impurity were compared and proposed.

CONCLUSIONThe method was rapid, sensitive and specific. It can be used for the identification of the isomeric purity in raw drug material of cefepime dihydrochloride.

Cephalosporins ; chemistry ; isolation & purification ; Chromatography, Liquid ; methods ; Drug Contamination ; Isomerism ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods

Natural hirudin extracted from the secretion of medical leech salivary gland is a single-chain peptide containing 65 aminoacid residues with molecular weight of 7000 D, and exists in three isomers of HV1, HV2 and HV3. Hirudin possesses three disulfide bridges forming the structure of core cyclic peptides, which binds to the catalytic site of thrombin so as to inhibit the catalysis of thrombin. Its c-terminus rich in acidic aminoacid residues possesses hydrophilicity, and is free on the molecular surface, and can bind with fibrin recognition site of hirudin. The minimal segment of 12 - 16 C-terminal acidic residues keeps the minimal activity of anti-thrombosis. Thus, hirudin, as a potent and specific inhibitor of thrombin, can be used to protect from and to treat clinically thrombosis. As it has some disadvantages such as short half-life, bleeding side-effect and mono-function, and so on, hirudin has been fused with some other functional proteins in recent years. The obtained fusion proteins can prolong the half life of hirudin, or relieve it bleeding side effect, or bring new functions, such as thrombolysis, inhibiting the platelet aggregation, targeting specifically. The research progress in hirudin fusion protein was summarized in this review.
Anticoagulants ; pharmacology ; Delayed-Action Preparations ; Drug Delivery Systems ; Glucokinase ; biosynthesis ; genetics ; pharmacology ; Hirudins ; biosynthesis ; genetics ; pharmacology ; Humans ; Platelet Aggregation Inhibitors ; pharmacology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Urokinase-Type Plasminogen Activator ; biosynthesis ; genetics ; pharmacology

BACKGROUNDTraumatic brain injury (TBI) is a major cause of death and disability in children and young adults worldwide. Therefore, we investigated the role of AG490 in regulating brain oedema, expression of CD40 and neurological function after TBI.

METHODSSprague Dawley rats (n = 240) were randomly divided into a sham operation group, TBI+saline group and TBI+AG490 (JAK/STAT inhibitor) group. Members of each group were euthanized at 6, 12, 24 or 72 hours after injury. Neurological severity score (NSS) was used to evaluate the severity of neurological damage. Brain water was quantitated by wet/dry weight method. The expression of CD40 was assessed by flow cytometry.

RESULTSIn both the TBI+saline group and the TBI+AG490 group, the brain water content was elevated after TBI, reached a peak at 24-hour and remained high for the rest of the period investigated; the expression of CD40 reached a peak 24 hours after TBI; the NSS was elevated after TBI and then decreased after 6 hours. Elevations in the level of CD40, degree of brain edema and NSS after TBI were significantly reduced in TBI+AG490 group.

CONCLUSIONInhibition of the JAK/STAT signalling pathway reduces brain oedema, decreases the expression of CD40 and exerts neuroprotective effects after TBI.

Animals ; Brain Edema ; metabolism ; Brain Injuries ; drug therapy ; CD40 Antigens ; analysis ; Flow Cytometry ; Janus Kinases ; metabolism ; Male ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; STAT Transcription Factors ; metabolism ; Tyrphostins ; therapeutic use

To explore the absorption mechanism of paeonol-beta-CD from various intestinal segments and offer biopharmaceutics data for paeonol new dosage form. The absorption kinetics and permeability rate consatants were investigated by the in situ perfusing method in rats. The absorption of the drug conforms to the firt-order kinetics and passive transport mechanism . The results indicate that paeonol-beta-CD absorption mechanism wasn't change.
Acetophenones ; pharmacokinetics ; Animals ; Intestinal Absorption ; physiology ; Intestines ; metabolism ; Kinetics ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To analyze risk factors and prognostic risk factors in aged patients with coronary heart disease (CHD).Methods:Clinical data of 412 aged patients,who were suspected of CHD in our hospital,were retrospec-tively analyzed.According to results of coronary angiography,they were divided into CHD group (n=283 )and non-CHD group (n=129);risk factors and prognostic risk factors for aged CHD patients were analyzed.Results:Compared with non-CHD group,there were significant rise in percentages of >80 years (13.95% vs.28.98%),di-abetes mellitus (DM,37.98% vs.66.78%)and smoking history (32.56% vs.59.72%)in CHD group,levels of to-tal cholesterol [TC,(5.06±1.53)mmol/L vs.(5.41±1.36)mmol/L],triglyceride [TG,(1.53±0.82)mmol/L vs.(2.02±0.95)mmol/L]and low density lipoprotein cholesterol [LDL-C,(2.05±0.86)mmol/L vs.(2.57 ±1.15)mmol/L],and significant reduction in high density lipoprotein cholesterol level [HDL-C, (1.65 ±0.52) mmol/L vs.(0.86±0.38)mmol/L]in CHD group,P <0.05 or <0.01. Multi-factor Logistic regression analysis indicated that age,DM,hypertension history,smoking,high total cholesterol (TC)and hypertriglyceridemia were independent risk factors for CHD (OR= 1.38 ~8.65,P <0.05 or <0.01 );while serum total bilirubin and high density lipoprotein cholesterol were independent protective factors for CHD (OR=0.224,P =0.002);multi-factor Cox model analysis indicated that smoking history,high TC and hypertension were risk factors affecting prognosis of CHD patients (RR= 1.563 ~ 2.315,P < 0.05 all).Conclusion:In clinic,attention should be paid to reduce or eliminaterelated risk factors of aged CHD patients to improve prognosis.