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

Objective:To investigate the mechanism of endoplasmic reticulum (ER) stress-induced chemoresistance to cisplatin in gastric cancer cells. Methods:ER stress models were established in both BGC823 and SGC7901 gastric cancer cells. The expression of GRP78, an ER stress marker, was examined by Western blot analysis. Moreover, whether ER stress can decrease the sensitivity of gastric cancer cells to cisplatin and activate P38 was explored by flow cytometry and Western blot analysis, respectively. Whether ER stress-induced chemoresistance to cisplatin can be abrogated by blocking P38 activity in gastric cancer was also elucidated using flow cytometry. Results:GRP78 protein expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with tunica-mycin (TM) or thapsigargin (TG) for 8, 16, and 24 h (P<0.05), compared with that in the group treated for 0 h. The apoptotic rates of TM-(or TG)-, cisplatin-, and TM (or TG) plus cisplatin-treated groups significantly increased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with the rate in the control group. The apoptotic rate of TM (or TG) plus cisplatin-treated group signifi-cantly decreased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with that of the cisplatin-treated group. Com-pared with the group treated for 0 h, phospho-P38 expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with TM (or TG) for 8, 16, and 24 h (P<0.05). No difference in P38 protein expression was observed between each group in both BGC823 and SGC7901 gastric cancer cells (P>0.05). Both P38 inhibitors, either SB203580 or PD169316, can inhibit the activation of P38. The inhibition of P38 activity can overcome ER stress-induced chemoresistance to cisplatin in gastric cancer cells (P<0.05). Con-clusion:ER stress can trigger the chemoresistance to cisplatin by activating P38 in gastric cancer cells.

Objective:To analyze the characteristics of health financing at the provincial level according to the total health expenditure since China health system reform began in 2009 and provide evidence for improving health fi-nancing policy. Methods:20 provinces were chosen and vertical and horizontal Comparative approach was used to an-alyze the data. Results:Total health expenditure increased for all regions, of which the biggest rate was Anhui prov-ince, about 82. 97%, while the largest increasing for government health care expenditure was Ningxia province, a-bout 108 . 71%. In 2012 , the provinces with social health expenditure share of total above 40% were allocated in the east region, and the number of provinces with out-of pocket payment share of total above 40% reduces to 5. Conclu-sion:Total health expenditure grew in all regions, but there were differences in the degree that this spending matched the economic level;The financing structure was optimized, but the characteristic of regional financing was different. Some provinces were under huge pressure to reduce out-of pocket payments. Suggestions: Under the premise of im-proving the funding level, financing structure adjustment must be focused, and public funding should play a bigger role and out-of pocket payments should be reduced.

Objective To explore the effects of gross saponins of Tribulus terrestris L.on inflammatory reaction and permeability of blood-brain barrier in rats following cerebral ischemia-reperfusion injury and their potential mechanisms. Methods Sixty SD rats were divided into sham operation group,model control group,gross saponins of Tribulus terrestris L.at low-dose (10 mg?kg-1 )and high-dose groups(30 mg?kg-1 ).Cerebral ischemia -reperfusion model was established with suture emboli method in middle cerebral artery of rats.Neural injury scores,the contents of Evans blue ( EB) and myeloperoxidase( MPO) activities in rat brain were measured 24 hours after the cerebral reperfusion post 2 h ischemia.Content of tumor necrosis factor-α (TNF-α) in rat brain was detected by ELISA; expression levels of matrix metalloproteinase-9(MMP-9) in rat brain was determined by Western blot. Results Compared to the model control group,the neurological deficit scores were significantly decreased(P<0.05),MPO activities and EB contents decreased(P< 0.05 or P< 0.01) in the treatment groups.The expression levels of TNF-α were significantly lower in the treatment groups(0.760±0.110) mg?g-1 and (0.670±0.073) mg?g-1 compared to (0.920±0.128) mg?g-1 in the model control group ( P< 0.05 or P< 0.01). The MMP-9 expression levels were (1.770± 0.181)% and(1.480±0.146)%,significantly lower than(2.200±0.186)% in the model control group(P<0.01). Conclusion Gross saponins of Tribulus terrestris L. exert neuroprotective effects on cerebral ischemia-reperfusion injury in rats through inhibiting the inflammatory reaction and decreasing the permeability of blood-brain barrier,which may be associated with the decrease of the TNF-α content and downregulation of the MMP-9 expression.

Objective To evaluate postprandial pharmacokinetics and bioequivalence of two preparations of cefdinir capsules in Chinese healthy volunteers. Methods In a two-way cross-over study, 24 healthy male volunteers were divided into two groups randomly and a single dose of cefdinir capsules of test and reference preparation were administered orally, respectively.The concentration in plasma was determined by LC-MS/MS. Pharmacokinetic parameters and bioequivalence were calculated and evaluated by DAS. Results The main pharmacokinetic parameters of test and reference were as follows: AUCt (4.35±1.09) μg??h??mL-1 and (4.12±1.22) μg??h??mL-1, AUC0-∞(4.53±1.12) and (4.53±1.73) μg??h??mL-1, t1/2 (1.74±0.29) h and (2.13±1.65) h, tmax(4.44±0.86) h and (4.54 ±1.16) h, Cmax(900±250) ng??mL-1 and (876±269) ng??mL-1 . Conclusion The test and reference preparation of cefdinir capsules are bioequivalent.

Objectives To explore the role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia-induced apoptosis of alveolar epithelial cells A549. Methods A549 cells were cultured in vitro and divided randomly into control (incubated with 5%CO2), hyperoxia group (exposed to a mixture of 900 ml/L O2 and 50 ml/L CO2 at speed of 3 L/min for 10 mins, then cultured in a closed environment) and LY333531 group (treated with 10μmol/L of PKCβinhibitor LY333531 for 24h then induced with hyperoxia for 10 mins). The cellular morphology was observed under inverted microscope at 12, 24 and 48 h of treatment. The cell apoptosis was detected by lfow cytometry. Expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were detected by immunohistochemistry after 24 h of treatment. Results Comparing to the control group, the cellular morphology of A549 in the hyperoxia group changed to spherical shapes and space between cells increased, the living cell count decreased and suspension cell increased. The living cell count in LY333531 group increased and suspension cell decreased than those in hyperoxia group but not reach the levels of the control group. The apoptosis rate of A549 cells and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 at 24 h were signiifcantly increased in the hyperoxia group than those in the control group, while the apoptosis rate and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were greatly decreased in the LY333531 group than those in the hyperoxia group (all P<0.01). Conclusions The expression of PKCβin A549 cells can be increased by the hyper-oxia induction but reduced by LY333531, and then the expressions of Pin1, p66Shc and p66Shc-Ser36 are reduced. Thus the re-duced apoptosis of A549 cells relieve the cell injury induced by hyperoxia.

Objective To find a way to measure and count plane distribution of cells distributed on single layer and compare differences of undifferentiated mesenchymal cells of periosteum germinal layer from different parts of the body.Methods After counting the number of undifferentiated mesenchymal cells of periosteum germinal layer from different parts of the body microscopically and figuring out the number of cells per area unit in each periosteum specimen,the obtained data were statistically analyzed and the stratum structure of periosteum observed microscopically.Results The homogeneity of variance test showed homoscedasticity,with no statistical significance(P＞0.05).The analysis of variance found homoscedasticity but showed no statistical significance(F=0.253,P＞0.05).The periosteum of patel- la,tibial plateau and costa had two layers,while the periosteum of costal cartilage had three layers. Conclusions There is no conspicuous difference upon proliferation and evoluting activities of periosteum from different parts of body.Therefore,it is unnecessary to choose specific parts for drawing the periote- um in clinical situation.In the meantime,the structure of periosteum from different parts diversifies.

Objective To explore the changes of tumor necrosis factor-?(TNF-?) in serum and cerebrospinal fluid(CSF) of children with acute lymphoblastic leukemia (ALL) and acute myelogenous leukemia(AML) and its clinical significance.Methods TNF-? in serum and CSF were measured by radioimmunoassay and CSF samples were obtained from 31 cases of childhood acute leukemia before treatment, on complete remission(CR), and continuous CR.Results Serum TNF-? was in ALL and AML before treatment [(24.35?4.84) pmol/L and(28.65?5.12) pmol/ L],which were significantly higher than those of healthy controls[(11.2 8? 1.69) pmol/L, P

Objective Acute pulmonary thromboembolism(PTE)patients with right ventricular dysfunction(RVD)may benefit from thrombolytic therapy but may end up with worse prognosis.RVD was assessed in prognosis to which a model on it was constructed to decide the indexes correlated to the best prognosis.Methods This prospective study included 520 consecutive acute PTE patients from 41 hospitals in China between June 2002 and February 2005.All the patients were evaluated by transthoracic echocardiography(TTE),CT pulmonary angiography(CTPA),laboratory tests,and blood gas analysis.Physicians were asked to record all the clinical manifestations.Data from Univariate analysis demonstrated the parameters correlated with an 14-day clinically adverse outcomes.Multiple logistic regression analysis was used to decide the independent predictors and to construct a prognostic model.Results Mcan age of the patients was 57.4±14.1 years and 323 of them(62.1%)were male.The 14-day mortality in normotensive patients with RVD was higher(2.0%vs0.4%,P＜0.01)than those without.ROC curve showed the best cut.ofr values of RVED/LVED and SPAP for 14-day prognosis were 0.67 and 60 mm Hg,respectively.SPAP independently predicted 3-month clinicaI outcomes(P＜0.01).Results from Univariate analysis demonstrated tllat 24 parameters were correlated with an adverse 14-day clinical outcomes.which include palpation,syncope,panic,cyanosis,respiratory rate≥30/min,pulse≥110/min,jugular vein,accentuation of P2,murmurs in tricuspid area,time interval from onset,creatine phosphokinase(CPK),lactate dehydrogenase (LDH).alveolar-arterial PO2 difierence(PA-aO2),white blood cell(WBC)＜4.0×109/L or＞10.0×109/L,platelet.thrombus on TTE,RV/LV＞1.1,TRPG＞30 mm Hg,IVCmin＜8 mm,RVD,RVED/LVED＞0.6,SPAP＞60 mm Hg.RVWM.PTE range larger than two lobes or seven segments on CTPA.Furthermore,a multiple logistic regression model implied 8 predictors including RVD,RVED/LVED＞0.6,SPAP＞60 mm Hg,pulse≥110/min,accentuation of P2,Syncope,CPK,WBC＜4.0×109/L or＞10.0×109/L be independent predictors of an 14-day clinically adverse outcome(P＜0.01).This model seemed to fit well (P＜0.001).Wle chose a cut-off value as P≥0.2 and compared the model to the original derivation samples.Data showed that the sensitivity(true positive rate)was 81.82%,specificity was 92.11%.false positive rate was 18.18%.coincidence was 91.14%.and the concordarlCe rate was 80.96%.Conclusion RVD seemed a nice discriminator for poor prognosis in norrnotensive patients.Early detection of RVD(especially RVED/LVED＞0.67 and/or SPAP＞60 mm Hg)was beneficial for identifying patients at high-risk and the multiple logistic regression model(P＜0.001)could be well fitted.

Objective:To explore the scientificity of the public hospital liability policy governance from the perspective of policy tools,and to provide evidence for the optimization and adjustment of subsequent policies.Methods:Policy texts at the national and regional levels from 2009-2022 were selected,and the coding results were statistically analyzed using Excel 2019 based on Rothwell and Zegveld's classification of policy tools.Results:A total of 128 policy texts were included in the analysis,with 29.96%,34.16%,and 35.88%of the policy tools in the policy instrument dimension being demand-based,supply-based,and environment-based,respectively.The system mechanism dimension focused primarily on compensation mechanisms.Conclusion:The use of the three policy instruments is mainly dominated by environmental policies,with a relative lack of policies from demand side.It is recommended that the use of policies from demand side could be strengthened and the internal structure of policy tools be optimized,to improve the consistency and coupling of policies.