This study aimed to investigate the immune adjuvant effect and mechanism induced by chitosan nanoparticles carrying pJME/GM-CSF. In this study, plasmid DNA (pJME/GM-CSF) was encapsulated in chitosan to prepare chitosan-pJME/GM-CSF nanoparticles using a complex coacervation process. Immunohistochemistry was used to detect the type of infiltrating cells at the site of intramuscular injection. The phenotype and functional changes of splenic DCs were measured by flow cytometry after different immunogens were injected intramuscularly. The killing activity of CTLs was assessed using the lactate dehydrogenase (LDH) release assay. The preparation of chitosan-pJME/GM-CSF nanoparticles matched the expected theoretical results. Our results also found that, after pJME/GM-CSF injection, the incoming cells were a mixture of macrophages, neutrophils, and immature DCs. Meanwhile, pJME/GM-CSF increased the expression of MHC class II molecules on splenic DCs, and enhanced their Ag capture and presentation functions. Cell-mediated immunity was induced by the vaccine. Furthermore, chitosan-pJME/GM-CSF nanoparticles outperformed the administration of standard pJME/GM-CSF in terms of DC recruitment, antigen processing and presentation, and vaccine enhancement. These findings reveal that chitosan could be used as delivery vector for DNA vaccine intramuscular immunizations, and enhance pJME/GM-CSF-induced cellular immune responses.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Chitosan ; administration & dosage ; immunology ; Dendritic Cells ; immunology ; virology ; Encephalitis Virus, Japanese ; genetics ; immunology ; Encephalitis, Japanese ; immunology ; prevention & control ; virology ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; administration & dosage ; genetics ; immunology ; Humans ; Immunity, Cellular ; Japanese Encephalitis Vaccines ; administration & dosage ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Nanoparticles ; administration & dosage ; Spleen ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; virology ; Vaccines, DNA ; administration & dosage ; genetics ; immunology

Panax notoginseng (PN) is one of the commonly used clinical medicines for cardiovascular diseases and possesses a variety of pharmacological effects. P. notoginseng saponins (PNS) are the most important bioactive components in PN. The purpose of this study was to explain the mechanism of PNS on molecular network level. 18 targets of the main medicinal ingredients of PNS were gained by virtual screening based on pharmacophores and data mining. A protein interaction network of PNS was constructed with 189 nodes and 721 interactions. By a graph theoretic clustering algorithm Molecular Complex Detection (MCODE), 14 modules were detected. Gene ontology (GO) enrichment analysis of the modules demonstrated that the roles of PNS played in cardiovascular disease related to multiple biological processes, which could represent the characteristics of traditional Chinese medicine (TCM) as a whole to regulate the disease. The results showed that the blood circulation and hemostasis efficacy of PN related with the biological processes such as positive regulation of cAMP metabolic and biosynthetic process, platelet activation and regulation of blood vessel size, regulation of T cell proliferation and differentiation and so on. Therefore, the module-based network analysis will be an effective method for better understanding TCM.
Drugs, Chinese Herbal ; chemistry ; Humans ; Panax notoginseng ; chemistry ; Protein Interaction Maps ; drug effects ; Proteins ; chemistry ; Saponins ; chemistry

Heat sensitive protein medicines are increasingly exhibiting their critical importance on treatment of various diseases at present. But their popularization and application meet a great challenge because of their heat instability. In the present study, insulin was taken as a heat sensitive protein medicine and amino acid as bio-protective agent in order to investigate if these amino acids can protect the insulin from losing its bioactivity due to desiccation. The experiment was performed by using replica exchange molecular simulation (REMD) method and Gromacs software with Gromos96 (53a6) force field. The REMD results indicated that these amino acids could protect the bioactive structure of insulin during desiccation. The configurations of the protected insulin were preserved very well. Those results proved that amino acid is a kind of good bioactive protective agent for the heat sensitive protein medicines.
Amino Acids ; chemistry ; Desiccation ; Drug Stability ; Insulin ; chemistry ; Molecular Dynamics Simulation ; Software

Objective:The response surface methodology (RSM) and Box-Behnken design were adopted to optimize the preparation for citrus oils β-cyclodextrin microspheres inclusion compound. And the physical characterization and heat stability were evaluated.Methods:The best preparation technology included inverse emulsion polymerization and saturated water solution method, with volatile oil weight ratio and microspheres, microspheres and water feeding ratio, inclusion temperature as impact factors, inclusion rate as the response value, establish regression model,. We explored the orange peel naphtha beta ring paste by microscopy, infrared spectroscopy (IR), differential scanning calorimetry (DSC) and heat stability test.Results:The best preparation technology included the essential oil with beta ring paste microspheres ( V: m), the ratio of 1:10, beta ring paste small ball and the ratio of water (m: V) for 1:15, and inclusion temperature for 41 ℃. The average encapsulation efficiency and the average rate of yield under optimized conditions were 62.21% and 85.24%, respectively. The physical characterization and thermal stability tests demonstrated that the β-cyclodextrin microsphere inclusion complex of volatile oil from Citrus was successfully prepared and the physical properties were stable. Conclusion:The preparation method of citrus oils β-cyclodextrin microspheres inclusion compound by using the response surface methodology.

OBJECTIVETo pool the data of studies about anticoagulation in non-massive pulmonary thromboembolism (PTE) and evaluate the efficacy and safety of low molecular weight heparin (LMWH) and unfractionated heparin (UFH) as the initial treatment.

METHODSMEDLINE CD-ROM from January 1966 to August 2003 and CBM CD-ROM from January 1978 to August 2003 were chosen for searching the randomized clinical trials (RCTs) that compared the efficacy or safety of LMWH and UFH in non-massive PTE. A meta-analysis was employed to evaluate the results of these two therapies.

RESULTSFive RCTs including 999 cases were analyzed. Compared with UFH, the combined odds ratio (OR) of LMWH in treating PTE was as follows: (1) The total OR of mortality of PTE patients treated with LMWH was 0.81 (95%CI 0.36-1.81, P > 0.05); (2) The total OR of recurrence of venous thromboembolism (VTE) in PTE patients treated with LMWH was 0.37 (95%CI 0.14-1.00, P=0.05); (3) The total OR of bleeding in LMWH was 0.47 (95%CI 0.16-1.39, P > 0.05);(4) The total OR of heparin-induced thrombocytopenia (HIT) in LMWH was 0.66 (95%CI 0.06-6.92, P > 0.05).

CONCLUSIONSLMWH and UFH can reduce the mortality and recurrence of VTE in patients with PTE in the same degree. The risk of major bleeding was similar in the two treatment groups. Initial subcutaneous therapy with the LMWH appeared to be as effective and safe as intravenous UFH in the initial treatment of PTE.

Adult ; Anticoagulants ; therapeutic use ; Female ; Heparin ; therapeutic use ; Heparin, Low-Molecular-Weight ; therapeutic use ; Humans ; Male ; Odds Ratio ; Pulmonary Embolism ; drug therapy ; mortality ; Randomized Controlled Trials as Topic ; Recurrence ; Thrombolytic Therapy ; Treatment Outcome

BACKGROUNDPulmonary endarterectomy is safe and effective surgical treatment for chronic thromboembolic pulmonary hypertension. This study aimed to evaluate the efficacy of pulmonary endarterectomy in treatment of thromboembolic pulmonary hypertension.

METHODSA retrospective study of 15 patients who underwent pulmonary endarterectomy in Beijing Chaoyang Hospital was performed. Obvious pulmonary hypertension and hypoxemia were observed in all patients. Bilateral pulmonary endarterectomy was performed under cardiopulmonary bypass with profound hypothermic circulatory arrest.

RESULTSTwo patients (2/15) died of residual postoperative pulmonary hypertension and bleeding complication. The other 13 cases had significant decrease in systolic pulmonary artery pressure ((92.8 +/- 27.4) mmHg vs. (49.3 +/- 18.6) mmHg) and pulmonary vascular resistance ((938.7 +/- 464.1) dynesxsxcm(-5) vs. (316.8 +/- 153.3) dynesxsxcm(-5)), great improvement in cardiac index ((2.31 +/- 0.69) Lxmin(-1)xm(-2) vs. (3.85 +/- 1.21) Lxmin(-1)xm(-2)), arterial oxygen saturation (0.67 +/- 0.11 vs. 0.96 +/- 0.22) and mixed venous O(2) saturation (0.52 +/- 0.12 vs. 0.74 +/- 0.16) postoperatively compared to preoperative data. Mid-term follow-up showed that the cardiac function of all cases returned to NYHA class I or II, with great improvement in 6-minute walking distance ((138 +/- 36) m) and quality of life.

CONCLUSIONSBilateral pulmonary endarterectomy using cardiopulmonary bypass with the aid of deep hypothermia and circulatory arrest can effectively reduce pulmonary hypertension and provide good mid-term hemodynamic and symptomatic results with low surgical mortality rate and few complications.

Adolescent ; Adult ; Echocardiography ; Endarterectomy ; adverse effects ; methods ; Female ; Hemodynamics ; Humans ; Hypertension, Pulmonary ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thromboembolism ; surgery ; Young Adult

Haizao Yuhu decoction (HYD) is a formula that has been used for approximately 500 years and famous for its efficiency in treating thyroid-related diseases in traditional Chinese medicine (TCM). HYD was first presented by Chen Shi-gong in a famous surgical monograph named Waike Zhengzong during the Ming Dynasty. We conducted the research to investigate the possible pharmacokinetic profile of different prescriptions of HYD in rats, in order to reveal the interactions of Haizao and Gancao drug pair with other herbs in HYD. Liquiritin, naringin, besperidin, peimine, peiminine liquiritigenin, glycyrrhizic acid, hergapten, nobiletin, osthole, glycyrrhetinic acid in blood samples were determined by UPLC-MS/MS. The result revealed tbat Haizao could enhance the peak concentration of glycyrrhizic acid. The other herbs in HYD may promote'the absorption of flavonoids in Gancao in normal rats, but inhibit the absorption of saponins and accelerate their metabolism. Gancao and Haizao drug pair could enhance the bioavailability of hesperidin, peimine, bergapten, nobiletin and osthole and prolong the elimination of peimine and naringin.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Magnetic Resonance Spectroscopy ; Male ; Mass Spectrometry ; Plasma ; chemistry ; Rats ; Rats, Wistar

Nowadays various protein medicines are increasingly playing significant roles in the treatment of many diseases, but the bioactive structures of such kinds of protein medicines are unstable because they are heat sensitive. Therefore, it is very important to explore a protective method and to explain the protective mechanism of protein medicines. In the present research, insulin was chosen as a heat-sensitive protein medicine, and a Group 3 late embryogenesis abundant (LEA) protein was chosen as its bioactive protectant during desiccation. The results of replica exchange molecular dynamics simulation suggest that comparing with insulin without any protection, the bioactive 3D structure and secondary structure of the insulin protected by LEA protein were preserved very well. All analyzing results proved that the LEA protein was a good bioactive protectant for heat sensitive protein medicines.
Animals ; Cold Temperature ; Drug Stability ; Freeze Drying ; Helminth Proteins ; chemistry ; Insulin ; chemistry ; Nematoda ; Plant Proteins ; pharmacology ; Protein Interaction Domains and Motifs ; Protein Structure, Secondary

Objective To investigate the correlation between ITPKB mutation's variant allele frequency (VAF) and prognosis of diffuse large B-cell lymphoma (DLBCL). Methods This study included 155 patients with DLBCL initially diagnosed in the People's Hospital of Xinjiang Uygur Autonomous Region from June 2014 to December 2020. Paraffin-embedded tumor tissue specimens were obtained, and tumor tissue DNA was extracted. A total of 475 hotspot genes including ITPKB were detected by the next generation sequencing to analyze the relationship of the VAF of high-frequency mutant gene with progression-free survival (PFS) and overall survival (OS). Results The mutation frequency of ITPKB was 18.71%. The PFS was significantly shorter in the patients with ITPKB mutations than in those without mutations (37 months vs. 108 months; HR=1.643, 95%CI: 0.920-2.934, P=0.093). The R-language based web tool was used to find the best VAF cutoff to differentiate prognosis. The patients were divided into two groups (VAF High vs. VAF Low+Wt) according to their VAF values. The optimal VAF threshold for ITPKB was 27.48% (HR=3.480, 95%CI: 1.70-7.13, P=0.00027). Multivariate Cox analysis was conducted using clinical indicators such as age, gender, COO classification, IPI, and LDH, and the results showed that PFS was associated with high ITPKB VAF (≥28%) (HR=3.592, 95%CI: 1.738-7.425, P < 0.001) which was an independent adverse predictor of PFS. Conclusion The high load of ITPKB mutation is an independent risk factor for the PFS of patients with DLBCL, and the VAF of ITPKB mutation has a prognostic predictive value for patients with DLBCL.

OBJECTIVETo determine the prevalence of beta-fibrinogen gene -455G/A, -148C/T polymorphisms in Chinese Han population and to investigate whether they were associated with pulmonary thromboembolism (PTE).

METHODSThe subjects consisted of 101 patients with PTE and 101 healthy controls matched with age and sex, from the same geographic area. All patients were diagnosed by high probability of lung ventilation/perfusion scan and/or multi-slice CT pulmonary angiography as well as medical history and clinical manifestations. Genome DNA was extracted from whole blood using KI-phenol-chloroform. Genotypes and allele frequencies of fibrinogen beta gene -455G/A, -148C/T polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Restriction enzyme HaeIII and HindIII digestion were used for detecting -455G/A, -148C/T polymorphisms respectively.

RESULTSRegarding fibrinogen beta gene -455G/A and -148C/, the allele frequencies G and A of fibrinogen beta -455 in the controls were 0.931, 0.069 while C and T of -148 were 0.777, 0.223 respectively, which were in good agreement with Hardy-Weinberg equilibrium. There was significant difference of -455G/A genotype frequencies distribution of AA, GA, GG between cases and in controls respectively, but no significant difference was found in the -148C/T polymorphisms. The frequencies of mutation allele -455A were 0.193, 0.169 in cases and in controls with P < 0.05 but there was no statistically significant difference of -148T allele. The presence of A allele of fibrinogen beta -455 was found to be a greater risk factor in cases than in controls. The odds ratio (OR) of GA and GA + AA were 3.723 (1.786 - 7.759), 3.749 (1.842 - 7.630), respectively. When compared with GG genotype, the P value was 0.0001.

CONCLUSIONThere was a complete linkage disequilibrium between fibrinogen beta -148C/T and -455G/A found. The frequencies of -455A, alleles in PTE disease were apparently higher than that of healthy adults but there was no difference in -148T alleles.

Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; Fibrinogen ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Linkage Disequilibrium ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pulmonary Embolism ; genetics ; Risk Factors