Abstract: Objective　To investigate the cause of poisoning leading to multi-organ damage in a patient with an unexplained condition, to confirm the type and exposure amount of the pathogenic factor, to identify the sources of pathogenic factors, and to provide references and bases for the clinical treatment and the prevention of such events. Methods　Starting with the unknown traditional Chinese medicine taken by patients for a long time, the targeted screening strategy was initially used to screen for alkaloid poisoning. Subsequently, a non-targeted screening strategy using Information-dependent Acquisition Mode (IDA) was used to screen the patient's blood, urine, and drug samples. Combining the toxicological effects of suspected compounds with the patient′s clinical manifestations, the main pathogenic factors were determined. Quantitative methods were established according to standard substances to quantify the pathogenic factors in all the samples. An epidemiological investigation was conducted to obtain the patient's medication history, and combined with the examination of the medication's specifications and content, the exposure dose of each pathogenic factor was determined. Finally, a Data data-independent acquisition (DIA) mode termed Sequential Window Acquisition of All Theoretical Fragment-Ion Spectra (SWATH) was used to screen all samples, unbiasedly collecting secondary mass spectrometry information of compounds, thereby verifying and supplementing the confirmed pathogenic factors. Results　Targeted screening ruled out common alkaloid poisoning. Tadalafil was detected in the patient's blood and urine. Tadalafil, sildenafil, chloropretadalafil, acetaminophen, and diclofenac were detected in unknown traditional Chinese medicine. The side effects of these compounds were consistent with the clinical manifestations of the patient. The highest daily average exposure doses of tadalafil, sildenafil, chloropretadalafil, acetaminophen, and diclofenac were 30.5 mg, 15.8 mg, 0.05 mg, 45.6 mg, and 3.2 mg respectively, which were seriously excessive. The SWATH mode also screened out the above five drugs. In addition, palmatine chloride was also detected. Conclusions　Combining the patient's clinical manifestations and epidemiological data, this study integrated targeted screening, untargeted screening, targeted quantitative strategies, data-dependent, and DIA mode to confirm that this case is a drug poisoning event caused by long-term overdose consumption of traditional Chinese medicine adulterated with chemical components. This study provides insights for the diagnosis and investigation of patients with poisoning for unknown causes, offering references for emergency detection and management of related poisoning incidents.

China ; Gastroesophageal Reflux ; therapy ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; methods

Objective:To optimize the formulation of HA-CA liposomes and to study the anti-tumor effect of HA-CA liposomes on uterine cervical carcinoma mice.Methods:The methods of preparing HA-CA liposomes were screened,and the optimal fomulation was selected by the orthogonal design experiment with the the phospholipids/cholesterol ratio,the drug/lipids ratio and the pH value of PBS buffer was 7.4 as entrapment enfficiency was the index.The release of HA-CA liposomes was studyed by dialysis bag method.Uterine cervical carcinoma cells were inoculated subcutaneously into right axillary ofBal b/c mice,after continuous treatment of 14 d,we weighed the tumor and calculated the rate of tumor growth inhibition.Results:HA-CA liposomes were prepared by thin film hydration methods.The optimal parameters were as follows:the phospholipids/cholesterol ratio was 4∶1,the drug/lipids ratio was 1∶ 30,the pH value of PBS buffer was 7.4.The release curve of HA-CA liposome and CA liposome was basically the same,both of which a sustained-release efficacy.The cumulative release of HA-CA liposomes and CA liposomes were 78.39％ and 83.01％ at 48h.The inhibition rate of HA-CA liposomes on U14 cervical cancer mice was 60.39％ and significantly higher than that of positive control group,which was higher than that of CA and CA liposomes.Conclusions:HA-CA liposomes can significantly inhibit the effect of U14 cervical cancer nude mice higher than that of CA and CA liposomes owe to the modification of the active target ligand HA.

Objective: Based on traditional Chinese medicine (TCM) inheritance system and integrated pharmacology platform for data mining of ulcerative colitis prescription, The difference of molecular mechanism between high-frequency herb combination and potential new prescriptions were screened to explore the potential molecular targets and signal pathways. Methods: The prescriptions of ulcerative colitis were collected from “Chinese medicine prescription dictionary”. Association rules and complex system entropy clustering were used to determine the frequency of usage and association rules of each drug, and identify high-frequency herb combinations and new potential prescriptions. By using integration platform, network construction and target prediction, the disease-target network, herb-target network, GO and KEGG pathway of ulcerative colitis related preparations of high frequency as well as potential new pairs were analyzed. The potential molecular mechanism of ulcerative colitis related targets was compared in high-frequency and new prescriptions by analysis of functions and pathways. Results: From 257 prescriptions, the frequency of usage and the association rules of prescription herb were analyzed. A total of six core combinations and three new potential prescriptions were identified, and the disease-target relationship obtained, and molecular mechanism differences between the high-frequency herb combination and the new potential prescription were compared. Conclusion: Combination of TCM inheritance system and integrated pharmacology platform can help to explore the TCM formula and potential molecular mechanism of ulcerative colitis treatment from a macro and micro perspective, and provide the guidance for the further research of classic prescription and its derivatives.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.
Enzyme Inhibitors ; pharmacology ; Free Radical Scavengers ; pharmacology ; High-Throughput Screening Assays ; Superoxides ; Uric Acid ; Xanthine ; Xanthine Oxidase ; antagonists & inhibitors

Lactoferrin (Lf) is one of the food protein belonged to the innate immune system. Apart from its main biological function of binding and transport of iron ions, lactoferrin also has many other functions and properties such as antibacterial, antiviral, antiparasitic, catalytic, anti-cancer, anti-allergic and radioprotecting. Lf is usually used as additives of food and cosmetics. The research of lactoferrin has been increasingly reported, and the application of lactoferrin as a drug carrier has drawn extensive attention over the recent year. In this paper, researches of lactoferrin as drug carriers are classified and summarized in brain targeting, liver tumor targeting, lung tumor targeting and oral delivery systems according to their different characteristics.
Administration, Oral ; Brain ; Drug Carriers ; Humans ; Lactoferrin ; chemistry ; Neoplasms

OBJECTIVETo investigate the effect of astragaloside (Astr), one of the active components of the Chinese medical herb Astragulus membranaceus, on cardiac fibrosis in chronic myocarditis and its relevant mechanisms.

METHODSEighty mice were randomized into 3 groups, the control group (n=20), the model group (n=30) and the Astr group (n=30). Mice in the model group and the Astr group were monthly intraperitoneally inoculated with CVB3, but to the control group equal amount of culture fluid was given instead. Mice in the control and the model group were fed with drinking water while those in the Astr group with drinking water containing Astr-sodium carboxymethycellulose at a concentration of 300 mg/L. All the survived mice were sacrificed 3 months later. Heart tissue of mice was stained by picrosirius red for calculating collagen volume fraction (CVF) with an automatic image analysis system. Expressions of transforming growth factor beta1 (TGF-beta1), platelet derived growth factor (PDGF), matrix metalloproteinase 1 (MMP-1), tissue inhibitor of metalloproteinase 1 (TIMP-1), MMP-13 and MMP-14 in heart tissue were detected by Western blot analysis.

RESULTSAs compared with the model group, in the Astr group, the mortality and CVF were significantly lower (53.3% vs. 23.3%, chi2 = 4.23, P < 0.05), and (17.4 +/- 1.2% vs. 8.6 +/- 0.9%, chi2 = 5.38, P < 0.05), respectively. As compared with the control group, Western blot analysis showed that expression of TGF-beta1 was decreased, MMP-1 and TIMP-1 were down-regulated, while expressions of MMP-13 and MMP-14 were up-regulated after Astr treatment.

CONCLUSIONAstr could lower the mortality and alleviate the myocardial fibrosis of mice with chronic myocarditis. Its antifibrotic effect might be realized by way of inhibiting TGF-beta1 expression and up-regulating the expressions of MMP-13 and MMP-14 in the heart tissues.

Animals ; Blotting, Western ; Chronic Disease ; Coxsackievirus Infections ; complications ; Drugs, Chinese Herbal ; therapeutic use ; Endomyocardial Fibrosis ; etiology ; metabolism ; prevention & control ; Male ; Matrix Metalloproteinase 13 ; metabolism ; Matrix Metalloproteinase 14 ; metabolism ; Mice ; Mice, Inbred BALB C ; Myocarditis ; complications ; drug therapy ; virology ; Random Allocation ; Saponins ; therapeutic use ; Transforming Growth Factor beta ; metabolism ; Triterpenes ; therapeutic use

BACKGROUND:Under the in vitro conditions of cell harvesting, culture, and transplantation, whether bone marrow stromal cells (BMSCs) can be effectively applied in local gene therapy remains unclear.OBJECTIVE: To construct a recombinant eukaryotic expression plasmid carrying human bone morphogenetic protein-7 (hBMP-7) gene, and to expect to enhance osteoinductive properties of rabbit BMSCs transfected.DESIGN, TIME AND SETTING: A cell-genomics in vitro observation was performed at the Laboratory of Scientific Research, Second Affiliated Hospital of Harbin Medical University between July 2006 and July 2007.MATERIALS: Human healthy fresh placental tissue was provided by the Department of Gynaecology and Obstetrics, Second Affiliated Hospital of Harbin Medical University. Written informed consent was obtained from the women. One healthy male New Zealand rabbit was provided by the Laboratory Animal Center, Harbin Medical University.METHODS: hBMP-7 gene was cloned from human placental tissue to construct a recombinant eukaryotic expression plasmid carrying hBMP-7 gene by conjugating with eukaryotic expression vector pcDNA3.1. BMSCs were isolated from rabbit bone marrow and cultured in vitro. Then they were divided into 3 groups: pcDNA3.1-hBMP-7-transfected, pcDNA3.1 -transfected, and untransfected. 5×106 BMSCs were inoculated into a 60 mm3 flask containing antibiotic-free medium 1 day prior to transfection.MAIN OUTCOME MEASURES: RT-PCR and immunohistochemistry were employed to detect hBMP-7 expression in BMSCs, alkaline phosphatase activity, hydroxypreline content, and osteocalcin production in each group. RESULTS: After 72-hour transfection, a 1.3 kb fragment was seen in the pcDNA3.1-hBMP-7-transfected group, showing brown granules in the endochylema, but not seen in the pcDNA3.14ransfected and untransfected groups. ALP activity in the pcDNA3.1-hBMP-7-transfected group significantly increased at 2 days after transfection, peeked at 8 days, and still increased at 10 days. At each time point, alkaline phosphatase activity, hydroxyproline content, and osteocalcin production were significantly higher in the pcDNA3.1-hBMP-7-transfected group than in the pcDNA3.1 -transfected and untransfected groups (P<0.05 or P<0.01).CONCLUSION: Recombinant eukaryotic expression vector pcDNA3.1- BMP-7 was constructed successfully. Results indicated that hBMP-7 was expressed in BMSCs sufficiently and was involved in inducing differentiation of BMSCs into osteoblasts. The method would provide substantial basement for hBMP-7 gene therapy.

Objective To investigate the prognostic factors of cervical high-grade squamous intraepithelial lesions treated by cold knife conization with negative margin.Methods Two hundred and sixty-six women with cervical high-grade squamous intraepithelial lesions treated by cold-knife conization with negative margins at Beijing Hospital between Jan 1999 and Jan 2004 were analyzed retrospectively.All patients were followed up with cytology,high-risk human papillomavirus(HPV)test and eolposcopy if necessary.Results The cervical CIN recurrence rate was 8.6% with no incidence of invasive cervical cancer after a median follow-up of 46 months.The recurrence was related to the grade of lesions and gland involvement pathologically.One of 20(5.0%)cases with cervical intraepithelial neoplasia(CIN)Ⅱ,9 of 164(5.5%)cases with CIN Ⅲ(excluding carcinoma in situ,CIS)and 13 of 82(15.8%)cases with CIS recurred(P

AIM:To make compound demethycantharidin(including Radix astragali and Rhizoma bletillae) into stomach remaining-floating sustained-release preparation. METHODS:Radix astragal and Rhizoma betillae pulverized to superfine powders were combined with hydrophilic gel to manufacture stomach remaining-floating sustained-release tablet(floating tablet). RESULTS:The tablet started to drift in 3 min in the artificial gastric juice and float lasted for more than 8 h. By isotopic tracering,the floating tablets given on an empty stomach were retarded for 2-3 h and on large meal for 4-5 h in combination with ordinary tablet only retarded for 1 h. A pharmacopoeia dissolution method was set up to determine the release. 50% of tablets were disintegration in 5 h,70% in 8 h. The t 1/2 was 5.1 h(2-4 h for ordinary tablets),and the MRT was 8.3 h(3.3 h for ordinary tablets),and the remaining-floating sustained-release tablets showed a relative bioavailability of 230.71%. CONCLUSION:Pretreatment of raw herbs from compound demethylcantharidin and selection of the excipients will help to produce the herb-containing floating tablet.