To investigate the effects of particle size, mPEG molecular weight, coating density and zeta potential of monomethoxyl poly(ethylene glycol)-poly(lactic-co-glycolic acid) (mPEG-PLGA) nanoparticles on their transportation across the rat nasal mucosa, mPEG-PLGA-NPs with different mPEG molecular weights (M(r) 1 000, 2 000) and coating density (0, 5%, 10%, 15%) and chitosan coated PLGA-NP, which loaded coumarin-6 as fluorescent marker, were prepared with the nanoprecipitation method and emulsion-solvent evaporation method, and determine their particle size, zeta potential, the efficiency of fluorescent labeling, in vitro leakage rate and the stability with the lysozyme were determined. The effects of physical and chemical properties on the transmucosal transport of the fluorescent nanoparticles were investigated by confocal laser scanning microscopy (CLSM). The result showed that the size of nanoparticles prepared with nanoprecipitation method varied between 120 and 200 nm; the size of nanoparticles prepared with emulsion-solvent evaporation method varied between 420 and 450 nm. Nanoparticles dispersed uniformly; the zeta potential of PLGA-NPs was negative; mPEG-PLGA-NPs was close to neutral; chitosan coated PLGA-NPs was positive; and the efficiency of fluorescent labeling were higher than 80%. In vitro leak was less than 5% within 4 h and nanoparticles were basically stable with lysozyme. The CLSM results show that the transportation efficiency of mPEG-PLGA-NPs with a high PEG coating density and high mPEG molecular weight was significantly higher than that of uncoated PLGA nanoparticles and also that of chitosan coated PLGA-NPs (P < 0.05). The hydrophilcity, zeta potential and particle size of nanoparticles play important roles on the efficiency of mPEG-PLGA nanoparticles to transport across the rat nasal mucosa.

Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .

Objective To confirm untoward reactions in uterine benign disease caused by the application of high intensity fo‐cused ultrasound (HIFU) is lighter .Methods Following‐up 877 cases of patients with uterine benign disease who received HIFU treatment from September 2014 to May 2016 in Zigong Fourth People′s Hospital ,the intra‐and post‐operative untoward reactions were summarized and analyzed .Results We summarized the occurrence of untoward reactions ,treatment measures and prognosis :98% of untoward reactions were SIR A to B level ,and fully recovered after treatment immediately to 1 week without special inter‐vention;2% of untoward reactions were SIR C to D ,also fully recovered after hospital and nursing treatment ;no serious complica‐tions(SIR D and above) occurred .Conclusion HIFU can be used in the treatment of uterine benign diseases in our hospital ,which could provide references for the treatment of uterine benign diseases by using HIFU .

Objective To study the anti-tumor effect and mechanism of bortezomib in primary acute leukemia cells from elderly patients.Methods Primary acute leukemia cells were treated with bortezomib 50-5000 nmol/L for 24-48 h,cell proliferation was analysed by MTT assay; apoptosis of primary acute leukemia cells was observed by fluorescence microscopy and flow cytometry; protein expression of bcl-2 and Bax was detected by Western blot.Results The cell viability was 90 ％ and 70 ％ when leukemia cells were treated with 50 and 5000 nmol/L bortezomib for 24 h,respectively.Meanwhile,cells showed (10.2±2.3) ％ and (13.3±3.3) ％ apoptosis.With prolonged treatment for 48 h,cell viability decreased to 86 ％ and 60 ％,respectively,while the apoptosis rates were increased to(18.4±3.9) ％ and(20.7±3.7) ％.Compared to the control group 0 nmol/L bortezomib,the differences were statistically significant (F =53.76,F =7.74,F =54.49,F =16.94,all P values ＜ 0.05).With the increase of bortezomib concentration,the bcl-2 protein expression was decreased,while Bax was up-regulated.Conclusion Bortezomib can inhibit primary leukemia cells from elderly patients proliferation and induce apoptosis.The mechanism may be associated with the changes in bcl-2 family protein expression.

Objective:To provide some thoughts for pharmaceutical treatment and care for the patients with pulmonary infection af-ter renal transplantation. Methods:Clinical pharmacists participated the whole treatment process of one case of pulmonary infection af-ter renal transplantation. According to the literatures combined with medical history, clinical symptoms and lab results, the drug treat-ment process of the patient was analyzed, and the key points of the optimized pharmaceutical care were summarized. Results: The pharmaceutical care included the dose adjustment of immunosuppressants at the early phase of the disease and after the improvement of clinical symptoms, attention paid to the interactions between multiple anti-infective drugs and immunosuppressive agents, dosage ad-justment based on the renal function of the patient, monitoring adverse drug reactions and drawing up personalized regimen. Conclu-sion:Through comprehensive medication monitoring, clinical pharmacists can help physicians develop timely and effective treatment programs and provide professional and effective pharmaceutical care for patients.

The present study is to establish Caco-2/HT29-MTX co-cultured cells and investigate the transport capability of PLGA nanoparticles with different surface chemical properties across Caco-2/HT29-MTX co-cultured cells. PLGA-NPs, mPEG-PLGA-NPs and chitosan coated PLGA-NPs were prepared by nanoprecipitation method using poly(lactic-co-glycolic acid) as carrier material with surface modified by methoxy poly(ethylene glycol) and chitosan. The particle size and zeta potential of nanoparticles were measured by dynamic light scattering. Coumarin 6 was used as a fluorescent marker in the transport of nanoparticles investigated by confocal laser scanning microscopy. The transport of furanodiene (FDE) loaded nanoparticles was quantitively determined by high performance liquid chromatography. Colchicine and nocodazole were used in the transport study to explore the involved endocytosis mechanisms of nanoparticles. Distribution of the tight junction proteins ZO-1 was also analyzed by immunofluorescence staining. The results showed that the nanoparticles dispersed uniformly. The zeta potential of PLGA-NPs was negative, the mPEG-PLGA-NPs was close to neutral and the CS-PLGA-NPs was positive. The entrapment efficiency of FDE in all nanoparticles was higher than 75%. The transport capability of mPEG-PLGA-NPs across Caco-2/HT29-MTX co-cultured cells was higher than that of PLGA-NPs and CS-PLGA-NPs. Colchicine and nocodazole could significantly decrease the transport amount of nanoparticles. mPEG-PLGA-NPs could obviously reduce the distribution of ZO-1 protein than PLGA-NPs and CS-PLGA-NPs. The transport mechanism of PLGA-NPs and mPEG-PLGA-NPs were indicated to be a combination of endocytosis and paracellular way, while CS-PLGA-NPs mainly relied on the endocytosis way. PEG coating could shield the surface charge and enhance the hydrophilicity of PLGA nanoparticles, which leads mPEG-PLGA-NPs to possess higher anti-adhesion activity. As a result, mPEG-PLGA-NPs could penetrate the mucus layer rapidly and transport across Caco-2/HT29-MTX co-cultured cells.

Objective: To explore the impact of situational sport games on gross movement and social development among children aged 4-5 years, and to provide references for promoting the healthy development of preschool children. Methods: Seventy one children aged 4-5 were randomly selected from one public kindergarten in Beijing during Apr. to Jul. of 2019 and were randomly divided into experimental group ( n =36, 8 week situational sports game intervention) and control group ( n =35, regular learning and playing). The Test of Gross Motor Development (TGMD-3) and social development scale for children aged 3-9 were used to assess gross movement and social development, respectively, which were then compared and analyzed. Results: Before intervention, the total score of gross motor development[intervention group (42.63±7.62), control group (43.00±9.16)] and social development[intervention group (211.88±6.68), control group (212.71±9.80)] showed no significant differences between the two groups( t =-0.18，-0.37， P >0.05). After intervention, increases in gross motor development[intervention group (67.80±9.80), control group (45.68±9.47)] and social development level[intervention group (228.30±6.37), control group (214.71±8.65)] were observed in both groups( t =25.82，3.22；15.90，2.13， P <0.05), with gross movement and social development scores higher in intervention group than that of the control group( t =9.66，7.54， P <0.05). Conclusion 8 week situational sports games can promote gross movement and social development of 4-5 year old children. It is suggested that interventions to promote early childhood development should be aware of multiple dimensions of development.

The lack of effective in vitro infection model for hepatitis E virus (HEV) has greatly hindered the quantitative analysis of neutralizing titers of anti-HEV antibodies and human sera, thus impeding further studies of HEV-stimulated antibody responses and the immunological mechanisms. In order to improve this situation, the infection of HepG2 cells that are inefficient for HEV replication was continuously monitored until the viral load reached the limit of detection on day 13, the results of which confirmed the feasibility of using this cell line to establish the infection model. Then, neutralization assays of five anti-HEV murine monoclonal antibodies and serum samples collected from four HEV vaccine recipients (collected before and after vaccination) were performed by 96 multi-channel parallel infections, nucleic acid extraction, and qPCR. The results showed that the cell model can be applied for quantitative evaluation of the neutralizing capacity of different antibodies and antiserum samples from HEV vaccine recipients. In this study, we have successfully established a high-throughput in vitro HEV replication model, which will prove to be useful for the evaluation of HEV vaccines and studies of HEV epitopes.
Animals ; Antibodies, Viral ; analysis ; immunology ; Hepatitis Antibodies ; analysis ; immunology ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; chemistry ; immunology ; physiology ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Neutralization Tests ; methods ; Virus Replication

Objective:To construct a chemiluminescense immune quantification assay based one paired mAbs against complexed prostate specific antigen ( c-PSA).Methods:Six week-old female BALB/c mice were immunized with the commercial c-PSA antigen.After serum titer reaching a platform stage ,the spleen was immunized and fused with mouse myeloma cell lines ( Sp2/0 ) .The hybridoma were screened by indirect ELISA ,and eight generated antibodies were paired to obtain a quantitative analysis of the chemical luminescence.Results:7D6 specifically recognized c-PSA,while 1A10 recognized total PSA(t-PSA).And the paired antibody 1A10/7D6 were determined to successfully construct a chemiluminescense immune response quantitative detection method through the detection of c-PSA standard and clinical serum samples .had,positive samples have statistically significant difference ( P<0.000 1 ) with negative samples.And the correlation coefficient R 2 was 0.97 between our c-PSA quantitative results and that of the Siemens c-PSA chemiluminescense immunoassay kit .The detection linear range was 0.1-100 ng/ml,and the sensitivity was 0.005 ng/ml.Conclusion:The paired monoclonal antibodies specifically detecting c-PSA were generated and a c-PSA chemiluminescense immunoassay were developed in this study .The detection capability of our method was comparable with that of the international commercial kit .

OBJECTIVETo probe the significance of specific IgG4 in sera of patients with cerebral cysticercosis for diagnosis and therapeutic evaluation.

METHODSSpecific IgG4 in sera of patients with cerebral cysticercosis was assessed using colloidal gold-labeled mouse-anti-human IgG4 McAb as probe. The results were compared with the CT image manifestation.

RESULTSThe specific IgG4 positive rate in sera of patients with cerebral cysticercosis was 97.8%, whereas sera from patients with other kinds of parasitosis or central nerve system disease and the control group were all negative, except for a weak cross-reaction of sera from patients with hepatic echinococoosis. The determination of specific IgG4 in sera of patients with cerebral cysticercosis during different times of treatment showed that along with an increase in treatment time and improvement of clinical symptoms, specific IgG4 level gradually decreased. The positive rate and intensity of specific IgG4 in sera from patients with cerebral cysticercosis were consistent with the number of cysticercus parasites in the brain and pathologic changes, such as survival, disintegration, death and calcification. Survival of cysticercus in the brain was objectively evaluated using this technique.

CONCLUSIONSThe determination of specific IgG4 in sera is a practical method for diagnosis and therapeutic evaluation of cerebral cysticercosis.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Specificity ; Humans ; Immunoglobulin G ; blood ; immunology ; Mice ; Mice, Inbred BALB C ; Neurocysticercosis ; blood ; diagnosis ; therapy ; Predictive Value of Tests ; Tomography, X-Ray Computed