AIM　To improve the treatment efficacy of anti-tumor drug mitoxantrone, the conjugation of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies were prepared. METHODS Mitoxantrone-loaded nanospheres were prepared with emulsion-heating solidification technique. A heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP), was used as the crosslinker of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies; pharmaceutical properties of immunonanocapsuls were studied; the conjugates of nanospheres and monoclonal antibodies was confirmed with immunological methods such as slide agglutination test, fluorescent immunossay and rosset formation test, fluorescent staining and scanning electron microscope. RESULTS　Mitoxantrone-loaded nanospheres were spherical, with smooth surface and median diameter of 0.665 micron. When stored at 3-5, 20-25 and 37℃, RH 75% for three months, the appearance, morphology, size distribution, drug loading and in vitro release characteristics showed no significant change and the stability was satisfactory. The size analysis demonstrated that there was no obvious increase in the particle size of nanoparticles after conjugation. Immunological tests indicate highly selective binding of antibody-targeted nanospheres to C-erbB-2-overexpressing cells SK-BR-3. CONCLUSION　The conjugation of mitoxantrone-loaded nanospheres and anti-C-erbB-2 monoclonal antibodies can keep the activity of anti-C-erbB-2 and increase the therapeutic efficacy of anti-mammary cancer drugs.

Objective To explore the protective effect of hesperidin pretreatment on concanavalin A (Con A)-induced acute liver injury and the effect on expression of TNF-α and IFN-γ. Methods Seventy-two SPF C57BL/ 6 mice were randomly divided into three groups: normal control group, model control group and hesperidin group. Acute liver injury model was established by injected with Con A. The hesperidin group was treated intragastrically with 1 000 mg·kg-1 hesperidin for 10 days. Model control group was treated intragastrically with the same volume of 0. 5% of sodium carboxymethyl cellulose. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase ( AST) were measured. Pathological changes in hepatic tissue were observed under microscope. The expression of TNF-α and IFN-γ mRNAs in hepatic tissue was measured by reverse transcription polymerase chain reaction ( RT-PCR). The contents of TNF-α and IFN-γ in serum were detected by ELISA. Results Compared with model control group, the contents of ALT and AST in serum were significantly decreased (P<0. 01) in hesperidin group. Pathological changes in hepatic tissue were markedly improved. The expression of TNF-α and IFN-γ in the hepatic tissues and serum were significantly downregulated (P<0. 01). The concentrations of TNF-α and IFN-γ in hesperidin group were (717. 05±205. 22) and(611. 06±92. 82)pg·mL-1 in 2 h,(811. 56±167. 47)and(786. 19±215. 44)pg·mL-1 in 6 h. Compared with model control group, the expressions of TNF-α and IFN-γ in the hesperidin group were significantly downregulated (P<0. 01). But there was no significant difference between hesperidin group and model control group in 6 h after treated with Con A(P>0. 05). Conclusion Hesperidin pretreatment protects mice from Con A-induced acute liver injury possibly by inhibiting the expression of TNF-α and IFN-γ in the liver of mice.

Objective To investigate the correlation between white matter hyperintensities (WMH)and hyperintense vessel sign (HVS) on fluid-attenuated inversion recovery (FLAIR) magnetic resonance imaging(MRI)in old adults and to explore the risk factors and pathogeneses of WMH.Methods We retrospectively collected imaging and clinical data of patients who had received both head and neck CTA and brain MRI within one month at our hospital from 2013 to 2016.The Fazekas visual scale was used to evaluate periventricular white matter hyperintensity(PWMH)and deep white matter hyperintensity(DWMH)in each brain hemisphere.According to the presence or absence of HVS in a cerebra[hemisphere,patients were assigned into an HVS-positive group or an HVS-negative group.Clinical data,PWMH,and DWMH differences were compared between the two groups.Results A total of 271 patients(542 cerebral hemispheres)were included in this study.HVS-positive imaging occurred in 79(14.6％)cerebral hemispheres and negative imaging was observed in 463 (85.4％) cerebral hemispheres.There was a significant difference between the HVS-positive and negative groups in the ipsilateral CIA stenosis(x2 =126.840,P＜0.01).The incidence of ipsilateral severe carotid artery stenosis in the HVS-positive group was 62.0％ (49/79),which was significantly higher than 9.9％ (46/463)in the HVS-negative group.The incidence of moderate-severe DWMH was 65.8％(52/79) in the HVS-positive group,which was higher than 34.8％ (161/463)in the negative group(x2 =34.962,P ＜20.01).Nevertheless,the incidences of moderate-severe PWMH in the two groups were 65.8％ (52/79) and 55.5％ (257/463),respectively,without a significant difference between them (x2 =6.944,P =0.074).After adjusting for age,gender,ipsilateral ICA stenosis,hypertension,diabetes,etc.multivariate analysis suggested that HVS-positive imaging was still an independent risk factor for DWMH(OR =2.653,95％CI:1.489-4.726,P =0.001).Conclusions HVS-positive imaging is an independent risk factor for DWMH in the elderly,but no clear correlation with PWMH is found.It suggests that hypoperfusion is a possible mechanism for the development of DWMH in the elderly.

Objective To establish a method for detection of aminophylline in blood samples of preterm infants . Methods A molecularly imprinted electrochemical sensing film on the glassy carbon electrode surface was prepared by electropolymerization using aminophylline as the template molecule and pyrrole as the functional monomer in 0.2 mol/L HAc-NaAc buffer solution ( pH 4.0).The surface morphology and properties of molecularly imprinted sensing films were characterized by three dimensional laser scanning microscopy , differential pulse voltammetry ( DPV) and electrochemical impedance spectroscopy ( EIS) while the effects of scanning cycle number and incubation time were investigated by square wave voltammetry(SWV) method in 5 mmol/L K3[Fe(CN)6] -0.1 mol/L KCl solution.Results Under optimized experimental conditions ,the SWV peak current difference was linear to the negative logarithm of aminophylline concentration in the range from 1.0 ×10 -7 to 1.0 ×10 -3mol/L with a detection limit (S/N=3) of 0.5 ×10 -8mol/L.The recovery rate was 92.2% -101.4%.Also, the molecularly imprinted electrochemical sensor for aminophylline had good selectivity , stability and reproducibility .Conclusion The molecularly imprinted electrochemical sensor for aminophylline can be used for rapid and accurate detection of clinical blood concentrations of aminophylline molecules in preterm infants in the future .

ObjectiveTo analyzing the psychological status of hospitalized patients of internal medicine and its impact factors.Methods Hospitalized patients of internal medicine underwent a survey by using Chinese Health Questionnaire (CHQ-12),Social Support Scale and Disease-Cognition Scale.Those with a score of ＞4 received further investigation of Symptom Check-List 90 (SCL-90).Correlation analysis was performed between all factors of SCL-90 and social support or disease-cognition scale score.Results There was no significant difference of psychological status between males and females ( P ＞0.05 ).All SCL90 factors were negatively correlated with social support,of which obsessive,paranoid,and phobic presented stronger negative correlations with social support and objective support (P ＜ 0.05 ).Furthermore,all factors were negatively correlated with disease-cognition scale score.A significantly negative correlation between phobic factor and disease-cognition scale score was identified ( P ＜ 0.05 ).Improvement was found in 26.2％ patients after intervention.Conclusion Patients tend to show unhealthy emotion when they are under the stress of hospitalization.Hospitalization support system may need to be improved and patients' cognition of disease should be increased.

A strain of yeast capable of hydrolyzing ethyl ester of racemic Ketoprofen with high enantioselectivity has been isolated from soil after two-step enrichment. The yeast was identified as Trichosporon brassicae. The process of growth and enzyme production was investigated. The catalytic performance of the resting cell of KET4 in kinetic resolution of Ketoprofen was also investigated. When the conversion of substrate reached 41% , enantiomeric excess of the (S) - Ketoprofen produced was 91 % , indicating a high enantiomeric ratio of 45.

OBJECTIVETo observe clinical curative effect of cake-separated moxibustion on impaired glucose regulation (IGR) and explore its action mechanism.

METHODSSixty cases were randomly divided into a simple lifestyle intervention group (control group) and a cake-separated moxibustion combined with lifestyle intervention group (observation group), 30 cases in each one. The control group was treated with lifestyle intervention. Based on lifestyle intervention, cake-separated moxibustion at Pishu (BL 20), Weishu (BL 21) and Yishu (EX-B 3) was applied to the observation group. Fast plasma glucose (FPG), two hours plasma glucose after oral glucose tolerance test (OGTT2hPG), fasting insulin (FINS), homa insulin resistance index (HOMA-IR), blood lipid, body mass index (BMI) and waist circumference (WC) were observed in the two groups before and after treatment.

RESULTSAfter treatment, the OGTT2hPG and FPG were both decreased significantly (both P<0.05) in the two groups, compared between the two groups, the differences of FPG [(0.41 +/- 0.42) mmol/L vs (0.05 +/- 0.08)mmol/L] and OGTT2hPG [(0.85 +/- 0.53)mmol/L vs (0.17 +/- 0.19)mmol/L] were both statistically significant. There were no significant changes in FINS, HOMA-IR, blood lipid, BMI and WC in the control group before and after treatment (all P>0.05), but FINS, HOMA-IR levels, triglycerides (TG), total cholest-erol (TC), low density lipoprotein (LDL-C), BMI and WC in the observation group were decreased obviously after treatment (all P<0.05), which had statistical differences between the two groups (all P<0.05).

CONCLUSIONThe cake-separated moxibustion combined with lifestyle intervention can obviously control blood glucose levels, improve insulin resistance and blood lipid levels, decrease BMI and WC.

Adult ; Aged ; Female ; Glucose ; metabolism ; Glucose Intolerance ; metabolism ; physiopathology ; therapy ; Humans ; Insulin ; Male ; Middle Aged ; Moxibustion ; Waist Circumference

OBJECTIVETo investigate the effect of hepatitis B virus (HBV) X protein (HBx) on host cell cycle and HBV replication using cultured primary mouse hepatocytes to gain further insights into the mechanism of HBx-mediated modulation of cell cycle.

METHODSPrimary cultured mouse hepatocytes were transfected with HBx-expressing (pHBV) or HBx-selected (pHBV triangle X) plasmids, which were generated with sequences of the HBV ayw subtype 1.2 and included the green fluorescent protein (GFP) reporter gene. The levels of cell cycle proteins (p16, cyclin D1, p21, cyclin E and cyclin A) were measured with Western blotting, and HBV DNA was analyzed by Southern blotting and real-time PCR.

RESULTSThe freshly isolated hepatocytes showed no significant differences in levels of cell cycle proteins. However, at 48 hours post-transfection, the levels of cyclin D1, p21 and cyclin E were significantly higher and the level of p16 was significantly lower in the pHBV-transfected hepatocytes than in the pHBV triangle X-transfected hepatocytes (t = 15.713, 22.897, 14.680, and -19.584, respectively, P less than 0.05). The level of cyclin A was not different between the two groups (t = 0.142, P more than 0.05). At 72 hours post-transfection, the level of HBV DNA was higher in pHBV-transfected hepatocytes (rcDNA: 3288.336+/-448.011; dslDNA: 6458.318+/-182.163; ssDNA: 2760.613+/-393.561) than in pHBV triangle X-transfected hepatocytes (rcDNA: 515.721+/-62.530; dslDNA: 2122.228+/-28.347; ssDNA: 1632.013+/-207.021) and in the blank (untransfected) control group (P less than 0.05). Real-time PCR analysis of HBV DNA copy number per cell confirmed these results, (pHBV-transfected: 987.50+/-47.80 vs. pHBV triangle X-transfected: 303.67+/-33.94; t = 20.203, P less than 0.05).

CONCLUSIONSThe HBx protein can affect the levels of cell cycle proteins, which may induce quiescent hepatocytes to enter the G1 phase of the cell cycle and stay in this phase instead of entering the S phase, thereby promoting HBV intracellular replication.

Animals ; Cell Cycle ; Cell Cycle Proteins ; genetics ; metabolism ; Cell Line ; Hepatocytes ; cytology ; virology ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; Trans-Activators ; genetics ; metabolism ; Transfection

Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism