In this study, the buffering capacity of amphiphilic pH-sensitivity copolymer poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate (PEOZ-CHMC) was evaluated. The ammonium sulfate gradient method was used to prepare doxorubicin hydrochloride (DOX x HCl)-loaded liposomes (DOX-L), and then the post-insertion method was used to prepare PEOZ-CHMC and polyethylene glycol-distearoyl phosphatidyl ethanolamine (PEG-DSPE) modified DOX x HCl-loaded liposomes (PEOZ-DOX-L and PEG-DOX-L). The physico-chemical properties, in vitro drugs release behavior, cellular toxicity and intracellular delivery of liposomes were evaluated, separately. The results showed that PEOZ-CHMC has a satisfactory buffering capacity. The sephadex G-50 column centrifugation method and dynamic light scattering were used to determine the encapsulation efficiency (EE) and particle size of liposomes. The EE and particle size of DOX-L were (97.3 ± 1.4) % and 120 nm, respectively, and the addition of PEOZ-CHMC or PEG-DSPE had no influence on EE and particle size. The zeta potentials of three kinds of liposomes were negative. The release behavior of various DOX liposomes in vitro was investigated by dialysis method. In phosphate buffer solution (PBS) at pH 7.4, DOX x HCl was released from PEOZ-DOX-L in a sustained manner. While in PBS at pH 5.0, the release rate of DOX x HCl from PEOZ-DOX-L increased significantly, which suggested DOX x HCl was released from PEOZ-DOX-L in a pH-dependent manner. The intracellular delivery of liposomes was investigated by confocal laser scanning microscopy (CLSM). The CLSM images indicated that PEOZ-DOX-L showed efficient intracellular trafficking including endosomal escape and release DOX x HCl into nucleus, as well as the DOX-L and PEG-DOX-L had no this effect. The cytotoxicity of liposomes against MCF-7 cells was detected by using MTT assay. The results showed that antiproliferative effects of PEOZ-DOX-L enhanced with pH value decreased, whereas DOX-L and PEG-DOX-L did not have any significant difference in inhibitions at different pH conditions. Therefore, the problems of the inhibition of cellular uptake of liposomes and the failed endosomal escape of pH-sensitive liposomes by PEG chain can be overcome by the pH-sensitive liposomes constructed by PEOZ-CHMC.

Aim To provide references for clinical trials dose and rational drug use by evaluating mitochondrial toxicity of bentysrepinine on HepG2 cells.Methods Mitochondrial toxicity of bentysrepinine on HepG2 cells was cmomprehensively evaluated by measuring proliferation inhibition rate, lactic acid content in culture supernatant, reactive oxygen species(ROS) content, mitochondrial membrane potential (MMP) variation and the activity of mitochondrial respiratory chain complex enzymes Ⅰ to Ⅳ.Results The half inhibitory concentration of bentysrepinine of HepG2 cells was 359 μmol·L-1.Compared with the control group, bentysrepinine could reduce the MMP, raise the level of lactic acid, increase the content of ROS and lower the activity of mitochondrial respiratory chain complex enzymes Ⅰ to Ⅲ with the concentration of 400 μmol·L-1(196 mg·L-1), showing an obvious mitochondrial toxicity.Compared with lamivudine and adefovir dipivoxil, bentysrepinine exerted no influence on indexes above with the same concentration 100 μmol·L-1.Conclusions Bentysrepinine shows an obvious mitochondrial toxicity on HepG2 cells with the concentration of 400 μmol·L-1.This mitochondrial toxicity is not presented with the concentration of 200 μmol·L-1.It shows that the safety range of bentysrepinine about mitochondrial toxicity is relatively wide.The test plays a guiding role in clinical trial dose design as well as clinical treatment.

In this study, the buffering capacity of amphiphilic pH-sensitivity copolymer poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate (PEOZ-CHMC) was evaluated. The ammonium sulfate gradient method was used to prepare doxorubicin hydrochloride (DOX x HCl)-loaded liposomes (DOX-L), and then the post-insertion method was used to prepare PEOZ-CHMC and polyethylene glycol-distearoyl phosphatidyl ethanolamine (PEG-DSPE) modified DOX x HCl-loaded liposomes (PEOZ-DOX-L and PEG-DOX-L). The physico-chemical properties, in vitro drugs release behavior, cellular toxicity and intracellular delivery of liposomes were evaluated, separately. The results showed that PEOZ-CHMC has a satisfactory buffering capacity. The sephadex G-50 column centrifugation method and dynamic light scattering were used to determine the encapsulation efficiency (EE) and particle size of liposomes. The EE and particle size of DOX-L were (97.3 ± 1.4) % and 120 nm, respectively, and the addition of PEOZ-CHMC or PEG-DSPE had no influence on EE and particle size. The zeta potentials of three kinds of liposomes were negative. The release behavior of various DOX liposomes in vitro was investigated by dialysis method. In phosphate buffer solution (PBS) at pH 7.4, DOX x HCl was released from PEOZ-DOX-L in a sustained manner. While in PBS at pH 5.0, the release rate of DOX x HCl from PEOZ-DOX-L increased significantly, which suggested DOX x HCl was released from PEOZ-DOX-L in a pH-dependent manner. The intracellular delivery of liposomes was investigated by confocal laser scanning microscopy (CLSM). The CLSM images indicated that PEOZ-DOX-L showed efficient intracellular trafficking including endosomal escape and release DOX x HCl into nucleus, as well as the DOX-L and PEG-DOX-L had no this effect. The cytotoxicity of liposomes against MCF-7 cells was detected by using MTT assay. The results showed that antiproliferative effects of PEOZ-DOX-L enhanced with pH value decreased, whereas DOX-L and PEG-DOX-L did not have any significant difference in inhibitions at different pH conditions. Therefore, the problems of the inhibition of cellular uptake of liposomes and the failed endosomal escape of pH-sensitive liposomes by PEG chain can be overcome by the pH-sensitive liposomes constructed by PEOZ-CHMC.
Cell Nucleus ; Doxorubicin ; analogs & derivatives ; chemistry ; Endosomes ; Formates ; chemistry ; Humans ; Liposomes ; chemistry ; MCF-7 Cells ; Microscopy, Confocal ; Particle Size ; Phosphatidylethanolamines ; Polyamines ; chemistry ; Polyethylene Glycols ; chemistry

Objective To explore the risk factors and clinicopathologic characteristics of severe hand,foot and mouth disease (HFMD),and to provide a scientific basis for clinical interventions.Methods The clinical data of 2606 hospitalized children with HFMD in Ningbo No.2 Hospital were analyzed retrospectively from January 2010 to December 2012.Clinical characteristics were compared between two groups (regular case group and severe case group),and risk factors of severe HFMD were analyzed by Logistic regression test.The clinicopathologic features were analyzed from an autopsy case.Results Among total 2606 cases,619 cases were severe HFMD (23.7％).Logistic regression test showed that EV71 infection,poor spirit,blood glucose ＞ 9 mmol/L,high fever (temperature ≥ 39.1 ℃) lasted more than three days,younger than 3 years and abnormal neural reflex were the independent risk factors of HFMD seriousness.The results of the autopsy showed that the central nervous system,lung and intestinal were most severely.Conclusion Attention should be paid to observe the condition changes in HFMD patients with EV71 infection,younger than 3 years,nervous system symptoms,blood glucose ＞ 9 mmol/L and high fever.Targeted clinical intervention is particularly important for improving cure rate,and the protection of intestinal function can't be ignored.

OBJECTIVETo establish a highly sensitive and specific assay to detect dengue virus (DENV) envelope protein domain III (EDIII) IgG antibody, and to explore its value in the diagnosis and seroepidemiological survey of dengue.

METHODSThe DENV EDIII IgG antibody capture ELISA was developed using the recombinant full-length DENV EDIII, which was prepared by Pichia yeast expression system as the capture antigen. The serum samples were collected from the same group of 35 DENV-1 patients of primary infection during disease period in 2006 and their follow-up phase in 2010; and the sensitivity of the assay was compared to that of the commercial Panbio DENV IgG ELISA.

RESULTSThe sensitivity of DENV EDIII IgG ELISA in detecting the serum samples from disease period and follow-up phase was 87% (20/23) and 94% (33/35), respectively; whereas the sensitivity of Panbio DENV IgG ELISA was 71% (25/35) and 0, respectively. The sensitivity of DENV EDIII IgG ELISA in detecting the serum samples from both periods was similar, without statistical significance (χ(2) = 0.946, P = 0.331). For serum samples from disease period, the sensitivity of DENV EDIII IgG ELISA was comparable with that of Panbio DENV IgG ELISA (χ(2) = 1.924, P = 0.165). However, DENV EDIII IgG ELISA demonstrated a significantly higher sensitivity than Panbio DENV IgG ELISA in detecting the serum samples from follow-up phase (χ(2) = 62.432, P = 0.000).

CONCLUSIONDENV EDIII IgG capture ELISA is highly sensitive in detecting IgG in the serum samples from either disease period or follow-up phase. This method might be a promising alternative for diagnosis and seroepidemiologic survey of dengue.

Antibodies, Viral ; blood ; Dengue ; diagnosis ; immunology ; virology ; Dengue Virus ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin G ; blood ; Protein Structure, Tertiary ; Sensitivity and Specificity ; Seroepidemiologic Studies ; Viral Envelope Proteins ; immunology

Objective: To investigate the relationship between P27，P53 and PCNA expression in human gastric carcinoma tissues and clinicopathological parameters. Methods: The expression of P27，P53 and PCNA in 62 human gastric carcinoma tissues was examined with immunohistochemistry SP method. Results: Positive rates of P27，P53 and PCNA expression were 37.1％, 40.4%，83.9％. P27 expression was related with Bormann type, infiltrative depth, lymph node and distant metastasis and clinical stage. P53 expression was related with sex of patients, distant metastasis and clinical stage. PCNA expression was related with age of patients and infiltrative depth of tumor. P27 positive expression group was higher than negative group as to 5-year survival. P27 expression was in reverse relation with PCNA expression. Conclusion: The expression of P27, P53 and PCNA may be regarded as an important marker in judging malignant degree of gastric carcinoma,distant metastasis and prognosis.

Objective:To identify age and gender standardized body mass index among children and adolescents and explore their associations with high blood pressure (HBP) in late adolescence.Methods:The current study was based on the Health Promotion Program for Children and Adolescents, school-based surveillance successively conducted from 2012 to 2019 in Suzhou, China. A total of 11 812 children and adolescents aged 16-18 years, who had ≥4 examination records during 2012-2018 and were also involved in a surveillance program in 2019, were included. Latent class growth mixture modeling was used to identify the BMI-Z trajectories in different genders, and multivariate logistic regression was used to analyze the associations between different BMI-Z trajectories and risk of HBP in late adolescence.Results:Six distinct BMI-Z trajectories were determined for both genders:thin, slightly thin,standard, declining, overweight, and obese. Compared with the regular group, the obesity group had 94.0% ( OR=1.94, 95% CI: 1.43-2.63) and 107.0% ( OR=2.07, 95% CI: 1.33-3.22) increased risk of developing HBP in late adolescence in boys and girls, respectively. However, a neutral association was found between the descending group and HBP in late adolescence. Conclusions:Persistent obesity in children may increase the risk of HBP in their late adolescence. If an obese child restores normal weight before late adolescence, the risk of HBP may reduce.

Objective To investigate the palliative care cognition of medical workers and provide evidence for medical worker' s vocational education and treatment of patients with end-stage.Methods 280medical workers from seven different hospitals in five provinces were surveyed with self-designed questionnaire of their palliative care cognition from January to March 2011.Results 280 questionnaires were handed out and 269 were recovered,with the recovery rate of 96.1％.The palliative care cognition of medical workers was affected by work type,age,working years,and working departments.Doctors and medical workers with age between 36 and 45 years old and working years more than 10 years had higher palliative care cognition,and the scores were respectively (25.81 ± 5.51),(26.88 ± 4.40) and (26.23 ± 4.92).Nurses,medical workers with age under 25 years old,working years less than 5 years and in surgical department had lower palliative care cognition,and the scores were respectively (22.25 ± 6.31),(23.68 ± 6.56),(23.57 ± 5.78),(22.55 ± 6.49).The differences were statistically significant (F =6.989,3.961,5.877,2.677,respectively; P ＜0.01).Conclusions Education of palliative care for medical workers,especially for nurses,is needed to provide high quality medical service for patients.

OBJECTIVETo explore the comprehensive program of integrated Chinese and western medicine in the treatment of cognitive impairment in earthquake brain injury.

METHODSThe multi-central randomized controlled trial was adopted. The qualified subjects were randomized into an acupuncture + rehabilitation group (38 cases) and a rehabilitation group (35 cases). In the acupuncture + rehabilitation group, acupuncture, hyperbaric oxygen (HBO) and cognitive rehabilitation training were combined as the comprehensive program of integrated Chinese and western medicine in the treatment. In the rehabilitation group, HBO and cognitive rehabilitation training were adopted. The efficacy and safety were assessed.

RESULTS(1) After treatment of 2 months, the intelligent state, cognitive function and activity of daily life of patients were improved in the both groups (all P < 0.01). (2) After treatment of 2 months, the score of MMSE and the score of activity of daily life were (24.11 +/- 4.08) and (75.45 +/- 13.95) in the acupuncture + rehabilitation group, which were more significant as compared with (17.05 +/- 43.84), (66.06 +/- 12.75) in the rehabilitation group, respectively (both P < 0.01). In 6-month follow-up visit after treatment, the cognitive function and activity of daily life were improved continuously in the acupuncture + rehabilitation group, which was more significant as compared with the rehabilitation group (P < 0.01, P < 0.05).

CONCLUSIONThe integrated Chinese and western medicine of acupuncture, HBO and cognitive rehabilitation training is safe and effective in the treatment of cognitive impairment in earthquake brain injury. The therapeutic effect is more advantageous as compared with the simple rehabilitation program of western medicine.

Activities of Daily Living ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Brain Injuries ; psychology ; rehabilitation ; therapy ; Cognition ; Earthquakes ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo determine whether U50488H, a selective agonist of kappa-opioid receptor, could induce biphasic (early and late) cardioprotection against myocardial ischemia/reperfusion injury and to explore the underlying mechanisms.

METHODSIsolated perfused rat hearts were subjected to 30 min of ischemia followed by 120 min reperfusion and the cardiac function was evaluated.

RESULTLeft ventricular end-diastolic pressure (LVEDP), left ventricular developed pressure (LVDP) and maximal velocity of contraction and relaxation (+/-dP/dtmax) were improved when U50488H was administered 1 or 24 h before ischemia (P<0.05). Myocardial infarct size, activities of creatine kinase (CK) and lactate dehydrogenase (LDH) in the coronary effluent were lower in the U50488H pretreatment group than those in the control group. Administration of a selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib abolished the late phase of cardioprotection produced by administration of U50488H 24 h before ischemia. Activities of CK and LDH in the coronary effluent were higher in U50488H and celecoxib co-pretreatment group than those in U50488H group. However, administration of celecoxib did not block the early phase of cardioprotection by 1 h treatment of U50488H before ischemia.

CONCLUSIONThe late (but not the early) phase of cardioprotection induced by kappa-opioid receptor agonist might be mediated by COX-2.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer ; pharmacology ; Animals ; Cardiotonic Agents ; pharmacology ; Creatine Kinase ; metabolism ; Cyclooxygenase 2 ; physiology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Male ; Myocardial Infarction ; enzymology ; pathology ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; agonists