Body fat of 12 male adults were measured by water displacement me-thod(density method) at every morning for 5 successive days. The standard deviation of single observation was 0.29kg calculated by mean residual lung volume method. It was significantly lower than the value (0.5kg) calculated by the ordinary method (p

OBJECTIVE To explore the antitumor effects of combined tanshinoneⅠ(TanⅠ),metformin (Met) and aspirin (Asp) on malignant melanoma in mice and the possible mechanisms. METHODS C57BL/6 mice were injected with 0.1 mL B16F10 cells(2.8×109L-1)to establish the subcutaneous trans-plantation tumor model at the right forelimbs axillary.Then,the mice were divided into 8 groups according to body mass,including model group, TanⅠgroup(20 mg·kg-1,ip),Asp group(210 mg·kg-1,orally in drinking water), Met group (70 mg·kg-1, orally in drinking water), Asp+Met group, TanⅠ+Asp group, TanⅠ+Met group and TanⅠ+Asp+Met group,10 mice in each group.Each mouse drank about 7 mL of water every day for a total of 18 d.The mouse body mass was measured every other day and the tumor diameter was calculated every day. The mice were sacrificed after treatment, the tumor mass was measured and the tumor inhibitory rates were counted. The histopathological changes of the liver and spleen were observed with HE staining. The percentage of lymphocytes in the tumor tissue such as CD8+T,CD4+T and Treg cells was detected by flow cytometry.Inflammatory factors such as interleukin-6 (IL-6),IL-1β and tumor necrosis factor-α (TNF-α) were detected by ELISA. RESULTS The body mass (including tumor mass)of mice in different groups increased during the experiment,but that of TanⅠ+Asp+Met group increased more slower than in model group(P<0.01).At the end of the experiment,no lesions were seen in any liver or spleen tissue by pathological observation,and the number of survivors was 8/10(model group),8/10(TanⅠgroup),7/10(Asp group),7/10(Met group),8/10(TanⅠ+Asp group), 8/10 (TanⅠ+Met group), 7/10 (Asp+Met group) and 5/10 (TanⅠ+Asp+ Met group), respectively. Compared with model group,there were no obvious changes in tumor volume or tumor mass in TanⅠ, Asp and Met groups and other two-two joint groups,but the tumor volume and tumor mass in TanⅠ+Asp+ Met group were significantly decreased (P<0.01, P<0.05), and the tumor inhibitory rate in this group was 46.2%.Compared with the model group,the percentage of CD8+T cells increased(P<0.05) in TanⅠ+Asp+Met group,but there were no significant changes in other groups.The contents of IL-6, IL-1β and TNF-α in tumor tissue of TanⅠ+Met group were much higher than in model group(P<0.01, P<0.05,P<0.05)and the content of IL-6 increased in TanⅠ+Asp+Met group(P<0.01).CONCLUSION Combination of TanⅠ,Asp and Met can effectively inhibit the growth of melanoma in mice,which may be related to the increasing percentage of CD8+T lymphocytes and IL-6 in tumor tissue.However there are possibly some side effects.

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

Objective To discuss the clinical efficiency of endoscopic submucosal dissection (ESD) in treatment of early gastric cancer (EGC) and precancerous lesions. Method Clinical data of 106 patients with EGC or precancerous lesions who received the treatment of ESD from June 2012 to June 2015 was collected. Then analyzing the treatment effect, complications, postoperative pathology and long-term efficacy of ESD. Results The overall en bloc resection rate was 100.0％, the mean operation time was (61.8 ± 17.3) min and the mean diameter of the lesions was (2.7 ± 1.3) cm. No endoscopic massive haemorrhage occurred; The incidence of perforation and postoperative delayed bleeding was 6.6％ and 5.7％ respectively, which were cured by endoscopic treatment and there was no surgical treatment. Postoperative pathological results showed high differentiated adenocarcinoma in 23 cases, moderately differentiated adenocarcinoma in 29 cases, poorly differentiated adenocarcinoma in 19 cases, signet ring cell carcinoma in 3 cases and high grade intraepithelial neoplasia in 32 cases. Among them, 7 cases with basal tumor invasion, and there were no margin positive cases. So the R0 resection rate was 93.4％ and the R1 removal rate was 6.6％. The 7 cases with R1 resection reached R0 resection after second endoscopic treatment. 5 cases recurred within 1 years after the operation, and the recurrence rate was 4.7％. Up to December 2016, 3 patients died, the median follow-up period was 34 months and the 3 year survival rate was 97.9％. Conclusion ESD is safe and feasible in the treatment of EGC and precancerous lesions with the advantages of less trauma, faster recovery, less complications and reliable curative effect. Its clinical efficiency is similar to surgery.

Hypertrophic cardiomyopathy(HCM)is the second commonest form of heart muscle disease affecting children and adolescents and is a leading cause of sudden death in young athletes. The aetiology of HCM is heterogeneous in the paediatric population,and includes inborn errors of metabolism,neuromuscular disorders and malformation syndromes.However,most cases of apparently idiopathic HCM in childhood are caused by mutations in cardiac sarcomere protein genes. Hypertrophic obstructive cardiomyopathy(HOCM) is not uncommon in children who don't respond to drug treatment;for such children,surgical treatment is the only solution.Previous routine surgical methods include modified Konno operation,modified extended Morrow operation and interventional alcohol ablation or radiofrequency ablation for pediatric patients. At present,the reports and large-scale clinical experience are rare.

OBJECTIVETo investigate the correlation between different Chinese medicine (CM) syndromes and variations in microcirculation in septic shock patients.

METHODSseventy Septic shock patients were divided into four groups: heat damaging qi-yin group (HDQY, 23 cases); yin exhaustion and yang collapse group (YEYC, 26 cases); excessive heat in Fu organ group (EHFO, 10 cases); and heat damaging nutrient-blood group (HDNB, 11 cases). Sublingual microcirculation parameters were observed by sidestream dark-field (SDF) imaging and scored by Acute Physiology and Chronic Health Evaluation II (APACHE II) and the Sequential Organ Failure Assessment (SOFA), and parameters of microcirculation perfusion variations and prognoses were analyzed.

RESULTSCompared with those with qi-yin heat damage, perfused vessel density (PVD) in other groups decreased dramatically (P<0.05), and APACHE II scores increased significantly (P<0.05). In addition, the recovery time was prolonged substantially (P<0.05), and the mixed venous oxygen saturation (SVO2) decreased (P<0.05). Blood lactic acid increased significantly (P<0.05), and the mixed SVO decreased (P<0.05), in the YEYC group. Compared with the thermal injury camp blood group, sublingual microcirculation parameter variations showed no obvious difference in the YEYC and EHFO groups (P>0.05). There were significant positive correlations between CM syndromes and APACHE II scoring in different groups (r=0.512, P<0.05). There were negative correlations between PVD and APACHE II scoring (r=-0.378, P=0.043), the proportion of perfused vessels (PPV) and APACHE II scoring (r=-0.472, P=0.008), as well as between the microvascular flow index (MFI) and APACHE II scoring (r=-0.424, P=0.023) in different patients.

CONCLUSIONSublingual microcirculation may serve as a clinical diagnostic parameter of the patient condition, as well as being a prognostic indicator.

Aged ; Dopamine ; therapeutic use ; Dose-Response Relationship, Drug ; Female ; Hemodynamics ; Humans ; Male ; Medicine, Chinese Traditional ; Microcirculation ; physiology ; Middle Aged ; Mouth Floor ; blood supply ; physiopathology ; Perfusion ; Shock, Septic ; blood ; drug therapy ; mortality ; physiopathology ; Syndrome

In this study, cepharanthine hydrochloride (CH) was tested for its potential ability to modulate the expression and function of P-glycoprotein (P-gp) in the multidrug-resistant human chronic myelogenous leukemia cell line K562/ADR. Cytotoxicity of adriamycin (ADR) alone or in combination with CH or verapamil (VER) in K562 and K562/ADR cells was determined by MTT assay. Based on flow cytometric technology, the effect of CH or VER on the uptake and efflux of rhodamine123 (Rho123) and the accumulation of ADR in these cells was detected by measuring Rho123 or ADR-associated mean fluorescence intensity (MFI). The effects of CH and VER on P-glycoprotein (P-gp) expression in K562 and K562/ADR cells were also measured using a flow cytometry with PE-conjugated P-glycoprotein antibody. The results show that CH significantly enhanced the sensitivity of K562/ADR cells to ADR, 4 micromol x L(-1) of CH enhanced the sensitivity of K562/ADR cells to ADR by 7.43 folds, the reversal activity was 3.19 times higher than that of verapamil. However, CH had no effect on drug-sensitive K562 cells (P < 0.05). CH increased Rho123 and ADR accumulation in a concentration-dependent manner (2-8 micromol x L(-1)) and inhibited the efflux of Rho123 from these cells, but did not affect the accumulation and efflux of Rho123 from the wild-type drug-sensitive K562 cells. The inhibition effect of CH on P-gp expression in K562/ADR cells is in a time- and concentration-dependent manner. The reversal activity of CH is possibility related to inhibition of P-gp function and expression, which lead to an increased intracellular accumulation of anticancer drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antibiotics, Antineoplastic ; metabolism ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Benzylisoquinolines ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Doxorubicin ; metabolism ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells ; Rhodamine 123 ; metabolism

OBJECTIVETo investigate the viral etiology in hospitalized children with acute lower respiratory tract infections (ALRTI) plus platelet disorders.

METHODSA total of 255 children with ALRTI plus platelet disorders and 442 children with ALRTI and normal platelets, all of whom were hospitalized between March 2010 and February 2011, were included in the study. Their nasopharyngeal aspirate samples were collected, and RT-PCR or PCR was performed to detect 14 viruses.

RESULTSOf 255 ALRTI patients with platelet disorders, thrombocytosis was found in 253 cases (99.2%) and thrombocytopenia in 2 cases (0.8%). Among ALRTI patients with platelet disorders, 173 (67.8%) were infected with at least one virus, with human rhinovirus as the most common one, followed by parainfluenza virus type 3 (PIV3) and respiratory syncytial virus (RSV). The detection rate of PIV3 in the abnormal platelet group was significantly higher than in the normal platelet group (P<0.05). In contrast, the detection rate of influenza virus B (IFVB) in the abonormal platelet group was significantly lower than in the normal platelet group (P<0.05). The age distribution showed significant difference between the abnormal and normal platelet groups (P<0.01). Platelet disorders were mainly found in children under one year of age (P<0.01).

CONCLUSIONSThrombocytosis is often found in children with ALRTI caused by viruses, especially PIV3, but infection with IFVB seldom causes platelet disorders. Hospitalized children with ALRTI under one year tend to develop platelet disorders.

Acute Disease ; Adolescent ; Age Factors ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Infections ; blood ; complications ; virology ; Thrombocytopenia ; etiology ; Thrombocytosis ; etiology

Toxoplasma gondii is a serious foodborne zoonosis,which can not only affect the development of animal husbandry and the safety of meat products,but also cause great harm to public health.Therefore,the prevention of toxoplasmosis is crucial.Toxoplasma gondii vaccine which is regarded as an important measure to prevent toxoplasmosis has significant value to both public health and economics.This paper mainly summarizes advances in the study of Toxoplasma gondii vaccine in order to provide references for the further development of it,so as to control the toxoplasmosis more effectively.

Objective To investigate the effect of Toll-like receptor 5(TLR5) on the biological characteristics of human umbilical cord-derived mesenchymal stem cells (MSCs).Methods The lentiviral vectors expressing TLR5 were constructed and MSCs were infected with related lentiviruses .The overexpression of TLR5 in MSCs was verified by FACS analysis, RT-qPCR and Western blotting before its influence on cell proliferation , surface antigens and cell differentiation was evaluated.The biological function of TLR5 was verified by detecting cytokines IL-6 and IL-8 expression after TLR5 was activated by its agonist CBLB502.Results The results showed that the expression of mRNA and protein of TLR5 was upregulated in MSCs after infection of lentivirus carrying TLR5, but their proliferation , immunophenotypes and differentiation capacity were not influenced . Moreover, IL-6 and IL-8 expression was significantly increased when MSCs-TLR5 were treated with CBLB502 ( P <0.001 ).Conclusion The biological characteristics of MSCs were not affected after infection of lentivirus/TLR5, and the expressed TLR5 was functional in MSCs , which could activate the downstream signaling pathways and then regulate immune response.