A sensitive, rapid, simple and economical ultra-performance liquid chromatography–tandem mass spectro-metric method (UPLC–MS/MS) was developed and validated for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma using gliquidone as internal standard (IS). Liquid-liquid extraction method with ethyl acetate was used for sample pre-treatment. The separation was performed on an Xtimate Phenyl column using isocratic mobile phase consisting of A (aqueous phase: 0.15% formic acid and 0.05% ammonium acetate)and B(organic phase:acetonitrile)(A:B=40:60,v/v).The flow rate was 0.25 mL/min and the total run time was 6 min. The multiple reaction monitoring (MRM) transitions, m/z 494.5→394.5 for imatinib, 488.7→401.5 for dasatinib, 530.7→289.5 for nilotinib and 528.5→403.4 for IS, were chosen to achieve high selectivity in the simultaneous analyses. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 2.6–5250.0 ng/mL for imatinib, 2.0–490.0 ng/mL for dasatinib,and 2.4–4700.0 ng/mL for nilotinib.The method showed acceptable results on sensitivity,specificity, recovery, precision, accuracy and stability tests. This UPLC–MS/MS assay was successfully used for human plasma samples analysis and no significant differences were found in imatinib steady-state trough concentra-tions among the SLC22A5?1889T>C or SLCO1B3 699G>A genotypes(P>0.05).This validated method can provide support for clinical therapeutic drug monitoring and pharmacokinetic investigations of these three tyrosine kinase inhibitors(TKIs).

The cDNA fragment of JEV prME gene was cloned into the baculovirus shuttle vector (bacmid) to construct a recombinant baculovirus vector, defined as AcBac-prME. Then the recombinant baculovirus Ac-prME was obtained by transfecting Sf9 cells with AcBac-prME. Western blot analysis and immunofluorescence results indicated that both prM and E proteins were efficiently expressed in Sf9 cells. Electron microscopy suggested that prME was assembled into JEV-VLPs. To further evaluate the potential of JEV-VLPs as vaccine, the mice were immunized with JEV-VLPs and then challenged with lethal JEV. The results of mice survival and pathological changes demonstrated that the JEV-VLPs performed complete protection against JEV-P3 strain and relieved pathological changes in the mice brain significant. This study suggest that JEV-VLPs would be a potential vaccine for Japanese encephalitis virus.
Animals ; Antibodies, Viral ; immunology ; Encephalitis Virus, Japanese ; genetics ; immunology ; Encephalitis, Japanese ; immunology ; prevention & control ; virology ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Sf9 Cells ; Vaccination ; Vaccines, Virus-Like Particle ; administration & dosage ; genetics ; immunology ; Viral Envelope Proteins ; administration & dosage ; genetics ; immunology

Objective:To detect the change of proportions of αβT lymphocytes in mesenteric lymph nodes of miR-7 knockdown (miR-7KD)mice,and preliminarily explore its importance.Methods: The volume,weight index and total cells number of mesenteric lymph node in miR-7KD mice were measured.The pathologic morphology change of mesenteric lymph nodes was observed by HE stai -ning.And the changes on proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice were analyzed by Flow cytometry.Results:Compared with those of WT (wild type) mice,the volume,weight index,and the total cells number of mesenteric lymph node were significantly increased ( P<0.05 ).Moreover ,the pathologic morphology was significantly changed.The proportion and numbers of T lymphocyte were significantly increased;however,the proportion of B lymphocyte were significantly decreased (P<0.05). Notably,the proportion and number of CD4+T cells and CD8+T cells were significantly increased (P<0.05).Meanwhile,CD62L+T cell proportion were vigorously reduced and CD 69+T cell and IFN-γ+T cell proportion were notably up-regulated,which belong to CD4+and CD8+T cell(P<0.05).Conclusion:There was significant influence in the proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice,suggesting that miR-7 might play an important role in the composition and function of lymphocytes in mesenteric lymph nodes.

OBJECTIVE:To observe therapeutic efficacy and safety of cefoperazone sodium and sulbactam sodium combined with Shenmai injection in the treatment of elderly stroke associated pneumonia(SAP). METHODS:A total of 84 SAP patients in Dazhou Municipal Central Hospital during Mar. 2016-Mar. 2017 were divided into control group(42 cases)and observation group (42 cases)according to random number table. Based on routine treatment,control group was additionally given Cefoperazone sodium and sulbactam sodium for injection 3 g added into 5% Glucose injection 100 mL,intravenously,twice a day;suitable antibiotics were selected according to the results of pathogenic examination and drug sensitivity test. Observation group was additionally given Shenmai injection 100 mL added into 5% Glucose injection 250 mL intravenously,once a day,on the basis of control group. Both groups were treated for consecutive 2 weeks. Clinical efficacies of 2 groups were observed,and the levels of T-lymphocyte subsets(CD4+,CD8+,CD4+/CD8+)and inflammatory markers(TNF-α,hs-CRP,PCT),length of hospital stay before and after treatment. The ADR was recorded. RESULTS:The total response rate(95.24%)and marked response rate (47.62%)of observation group were significantly higher than those of control group(83.33%,23.81%);length of hospital stay in observation group [(15.24±3.53)d] was significantly shorter than control group [(18.43±4.21)d],with statistical significance (P<0.05).After treatment,the levels of CD4+and CD4+/CD8+in 2 groups were significantly higher than before treatment,and the observation group was significantly higher than the control group;the levels of CD8 +,TNF-α,hs-CRP and PCT in 2 groups were significantly lower than before treatment;the observation group was significantly lower than the control group,with statistical significance(P<0.05);there was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Based on routine treatment,cefoperazone sodium and sulbactam sodium combined with Shenmai injection treat SAP significantly,can effectively improve immune function, reduce inflammation level and shorten the length of hospital stay without increasing the occurrence of ADR.

Objective:To evaluate the safety and feasibility of robotic-assisted laparoscopic resection of bladder rhabdomyosarcoma.Methods:A retrospective study was performed in 4 children underwent robotic-assisted surgery for bladder rhabdomyosarcoma from July 2018 to September 2019 in our hospital. The perioperative information and short time outcomes were collected. All the four patients were male. The average age of these patients was 68 months (from 11 to 122 months). After cystoscopy biopsy was conducted, preoperative chemotherapy was carried out. Preoperative radiotherapy was also added depends on the tumor response, then robotic-assisted surgery was performed. Radical cystectomy and orthotopic neobladder reconstruction with bilateral ureter reimplanataion were carried out for the bladder was widely occupied case. Bladder preserving surgery was performed for the other three localized cases.Results:All of the four operations have been conducted successfully without conservation to open surgery. The mean operative time was 189(104-316) min while the mean estimated blood loss was 32.5(20-50) ml. The mean tumor diameter was 4.48(2.7-6.0)cm. According to the pathological results, all of the four cases were diagnosed as embryonal rhabdomyosarcoma. One case had positive margin while other three cases had negative margins. No complication more than Clavien-Dindo grade Ⅲ was observed. The mean length of hospital stay was 18(14-24)days and the mean follow-up time was 14.7(7-21) months. Postoperative chemotherapy was carried out according to the protocol of IRS low risk group for all cases (VAC, eight cycles). Postoperative radiotherapy was conducted for the case with positive margin. No recurrence was recorded during the follow-up period.Conclusions:Robotic-assisted laparoscopic technique for the treatment of bladder rhabdomyosarcoma in children is safe and feasible.

ObjectiveThis study aimed to understand the epidemiological characteristics of hand-foot-mouth disease （HFMD） in Minhang District， Shanghai from 2009 to 2020， and provide a scientific basis for the prevention and control of HFMD. MethodsThe case information of HFMD was collected from the National Notifiable Infectious Diseases Reporting System of Chinese Center for Disease Control and Prevention. We used descriptive epidemiological methods to analyze the population characteristics， temporal and spatial distribution of HFMD， the pathogen composition of the case and its changing trend. ResultsFrom 2009 to 2020， a total of 66，198 cases of HFMD were reported in Minhang District， Shanghai， including 377 severe cases （severe case rate 0.57%） and 3 deaths （severs case fatality rate 0.80%）. There were more cases of HFMD in boys than in girls （1.5∶1）. HFMD patients aged under 5 years predominated， accounting for 88.91% of all cases. Majority of the cases （91.42%） were in scattered children （55.80%） and children in kindergartens （35.62%）. The incidence showed a cyclical trend， with low incidence years and high incidence years appearing alternately. The peak period was from April to July， and sometimes there were small peaks during October to December. A total of 12 years time-space scanning analysis revealed 3 clusters. The cluster centers were located in Wujing Town， Huacao Town and Xinzhuang Town， respectively. The proportion of EV71 in common cases was generally decreasing， and reduced to zero in 2019. The proportion of CoxA6 had increased year by year， and reached 75.00% in 2020. CoxA6 became the dominant pathogen in recent years. The number of severe cases had decreased year by year since 2010， and the dominant pathogen was EV71 （90.03% on average） in severe cases. ConclusionThe incidence of HFMD in Minhang District of Shanghai has a downward trend from 2014. The dominant pathogen changes from EV71 to CoxA6， and the dominant pathogen in severe cases is EV71. The discovered temporal and spatial clustering pattern is helpful for in-depth understanding of the distribution and epidemic trend of HFMD in Minhang District， and provides a scientific basis for epidemic prevention and control.