Objective:To investigate impacts of usnic acid(UA)on malignant behavior of gastric cancer cells by regulating the chemokine(C-C motif)ligand 2(CCL2)-CCL2 receptor(CCR2)signal axis.Methods:SGC-7901 cells,a well growing human gastric cancer cell line,were treated with different concentrations of UA,which were grouped into low concentration(UA-L)group(62.5 μmol/L UA),medium concentration(UA-M)group(125 μmol/L UA)and high concentration(UA-H)group(250 μmol/L UA);meantime,the cells were transfected with CCL2 overexpression vector(pc DNA3.1 CCL2),empty vector(pc DNA3.1),silenced CCL2(si CCL2)and negative control(si control),and SGC-7901 cells were treated with 250 μmol/L UA,labeled as UA-H+pc DNA3.1 CCL2 group,UA-H+pc DNA3.1 group,UA-H+si control group and UA-H+si CCL2 group,another untreated SGC-7901 cells were taken as the control group.Flow cytometry,MTT and qRT-PCR were applied to detect cell apoptosis,proliferation,and expres-sion levels of CCL2 and CCR2 mRNA;Western blot was applied to detect expression levels of PD-L1,apoptotic protein(Bax),proli-ferative protein(CyclinD1,CCL2,CCR2)and immune escape related protein(B7H1);after co-culturing with CD8+T cells isolated and cultured in vitro,ELISA was applied to detect levels of IL-4,IFN-γ and IL-10 in the supernatant.Gastric cancer cells in each group were co-cultured with activated peripheral blood mononuclear cell(PBMC)1∶1 for 72 hours,and the sensitivity of gastric can-cer cells in each group to T-cell-mediated killing was compared.Results:Compared with control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-L group,UA-M group and UA-H group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels,Bax protein expression were obviously increased(P<0.05);compared with UA-H+pc DNA3.1 group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+pc DNA3.1 CCL2 group were obviously increased,while apoptosis rate,IL-4 and IFN-γ levels,and Bax protein ex-pression were obviously reduced(P<0.05);compared with UA-H+si control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+si CCL2 group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels and Bax protein expression were obviously increased(P<0.05).Conclusion:UA can inhibit gastric cancer cells proliferation,immune escape,and induce apoptosis,which may be related to the inhibition of the CCL2-CCR2 signaling axis.

The comparison between traditional Chinese medicine Jinzhen oral liquid (JZOL) and Western medicine in treating children with acute bronchitis (AB) showed encouraging outcomes. This trial evaluated the efficacy and safety of the JZOL for improving cough and expectoration in children with AB. 480 children were randomly assigned to take JZOL or ambroxol hydrochloride and clenbuterol hydrochloride oral solution for 7 days. The primary outcome was time-to-cough resolution. The median time-to-cough resolution in both groups was 5.0 days and the antitussive onset median time was only 1 day. This randomized controlled trial showed that JZOL was not inferior to cough suppressant and phlegm resolving western medicine in treating cough and sputum and could comprehensively treat respiratory and systemic discomfort symptoms. Combined with clinical trials, the mechanism of JZOL against AB was uncovered by network target analysis, it was found that the pathways in TRP channels like IL-1β/IL1R/TRPV1/TRPA1, NGF/TrkA/TRPV1/TRPA1, and PGE2/EP/PKA/TRPV1/TRPA1 might play important roles. Animal experiments further confirmed that inflammation and the immune regulatory effect of JZOL in the treatment of AB were of vital importance and TRP channels were the key mechanism of action.

Objective: To analyze the incidence and mutation characteristics of FLT3 gene mutation and clinical efficacy of tyrosine kinase inhibitor (TKI) in patients with mixed phenotype acute leukemia (MPAL). Methods: A total of 48 patients with MPAL who were admitted to Hebei Yanda Lu Daopei Hospital from June 2015 to February 2018 were retrospectively analyzed. The common mutated 58 genes in hematologic malignancies were detected by using amplicon-targeted next generation sequencing, of which internal tandem duplication (ITD) and point mutation occurred in the hotspot region of exon 14, 15 and 20 in FLT3 gene. Multiplex polymerase chain reaction (PCR) analysis was used to detect 35 gene fusions in hematological neoplams. Results: There were 7 cases of FLT3 mutation in 48 MPAL patients, which were all ITD mutations. The median length of the inserts of FLT3-ITD was 48 bp, and one MPAL patient carried 2 multiple length inserts simultaneously, and the median variant allele frequency (VAF) was 40.5% (7.9%-84.7%). There were no statistically significant differences in clinical and genetic characteristics between FLT3 mutation-positive and FLT3 mutation-negative MPAL patients (both P > 0.05). Among 7 FLT3 mutation-positive MPAL patients, 4 cases were often accompanied with RUNX1 mutation. A total of 4 MPAL patients with FLT3-ITD-positive received sorafenib or sunitinib combined chemotherapy, and 3 of them achieved complete remission. Conclusions ITD mutation is the main part in the FLT3 mutation of MPAL patients. FLT3-ITD-positive MPAL patients are often accompanied with RUNX1 mutation, which may benefit from targeted therapy with FLT3 kinase inhibitor.

Objective:To provide reference for clinical case diagnosis and treatment applications using descriptive analysis of patient-specific IgM and IgG test result of novel coronavirus pneumonia (COVID-19).Methods:Chemical luminescence was used to detect the levels of 423 confirmed or suspected cases IgM and IgG antibody, and the test result of patients with different clinical characteristics were compared and analyzed.Results:The positive rates of confirmed cases of COVID-19 were 80.4% (314/388) and 98.2% (381/388), for IgM and IgG respectively, and the positive rates of COVID-19 suspected cases specific IgM and IgG were 0.0% (0/24) and 45.8% (11/24), respectively. In patients at 6 weeks or more after the onset of the disease, the positive rate of IgG antibody was 100%. The level of IgG titer was generally higher in cases of 5~8 weeks after onset (mean: 112.70 AU/ml) than in cases of 1-4 weeks after onset (mean: 85.01 AU/ml) (U=8 531, P<0.0001). The level of IgG titer was higher in severe type cases than that of patients with ordinary type illness, with an average of 137.61 AU/ml. Conclusions:Specific IgM and IgG antibodies have strong application value in COVID-19 case diagnosis, and it is recommended to strengthen the tracking and monitoring of IgM and IgG titer levels in patients.

Objective To investigate epidemiologic features of genital Chlamydia trachomatis (Ct) infection in China,and to provide scientific evidence for developing control strategies.Methods A descriptive epidemiological study was conducted to analyze personal,spacial and temporal distribution of genital Ct infection cases reported from 105 national sexually transmitted disease (STD) surveillance sites in China between 2008 and 2015.Results The reported incidence rate of genital Ct infection increased from 32.48 per 100 000 in 2008 to 37.18 per 100 000 in 2015,with the average annual rate of increase being 1.95％.There was marked variation in the reported incidence of genital Ct infection (range,＜ 1/100 000-615.99/100 000) among different STD surveillance sites,which was relatively high in the Pearl River Delta region,Yangtze River Delta region,Minjiang River region and some ethnic minority areas in western China,but low in north China and central China.In addition,no cases were reported at a few sites in rural areas.The reported annual incidence rate was higher in females than in males from 2008 to 2015,and the ratio of male to female cases decreased from 0.61:1 in 2008 to 0.46:1 in 2015.Among all age groups,the sexually active population aged 20-44 years showed high reported incidence of genital Ct infection,with the highest reported incidence observed in the group aged 25-29 years (range,116.72/100 000-142.98/100 000);the group aged 15-19 years showed the highest average annual rate of increase (10.06％) in the reported incidence of genital Ct infection.Among all kinds of medical institutions,general hospitals had the largest number of reported cases (range,66.00％-74.22％),followed by gynecology hospitals,Maternal and Child Health (MCH) hospitals and specialized dermatovenereology hospitals.Conclusions Genital Ct infection has been one of important public health issues in China,more attention should be paid to its prevention,and effective preventive measures are needed to be developed according to epidemiologic features.

Objective To investigate epidemiological characteristics and trends of gonorrhea in China, and to provide a scientific basis for the development of control strategies. Methods An epidemiological study was performed on gonorrhea cases reported from 31 provinces, autonomous regions or municipalities directly under the central government between 2000 and 2014. Results The reported incidence rate of gonorrhea decreased from 22.92 per 100 000 in 2000 to 7.25 per 100 000 in 2014, with the average annual rate of decrease being 7.89%. There was a significant difference in the incidence of gonorrhea between different regions. The regions with the highest incidence rate of gonorrhea were Yangtze River Delta region(Zhejiang, Shanghai, Jiangsu)and Zhujiang River Delta region(Guangdong, Guangxi, Hainan), followed by northwest China (Xinjiang, Ningxia, Inner Mongolia). The incidence of gonorrhea was higher in males than in females, and the average male/female ratio increased from 1.96 ∶ 1 in 2000 to 4.52 ∶ 1 in 2014. The population aged 20 - 44 years showed high incidence of gonorrhea, and the highest incidence rate of gonorrhea was observed in the age group 25 - 29 years in both men and women. The incidence of gonorrhea decreased in all the age groups from 2000 to 2014 except the age group 15 - 19 years with an annual growth rate of 4.18%. Of 20 occupations, peasant workers accounted for the highest proportion (26.00%)of reported gonorrhea cases, and the number of reported gonorrhea cases showed a decreasing trend in all the occupations. Conclusions Gonorrhea remains a major public health issue in China, and effective measures based on epidemiological features are urgently needed to control gonorrhea.

Objective To investigate the trends in and characteristics of syphilis epidemiology in China,and to provide a scientific basis for the development of control strategies.Methods An epidemiological study was performed on syphilis cases reported from 31 provinces,autonomous regions and municipalities between 2000 and 2013.Results The reported syphilis incidence increased yearly from 6.43 per 100 000 person-years in 2000 to 32.86 per 100 000 person-years in 2013,with an average annual growth rate of 13.37％.Syphilis was reported in most (99％) counties/districts in China mainland except for a few remote mountain areas,with a significant difference in the incidence of syphilis between different regions.The regions with a high incidence of syphilis included minority areas in Northwest China (Xinjiang,Qinghai and Ningxia),Minjiang River area,Yangtze River Delta (Zhejiang,Shanghai),Zhujiang River Delta (Guangxi,Guangdong),etc.There had been a rapid increase in the incidence of syphilis in some regions previously reported to have a low incidence (such as Guizhou,Shaanxi,Henan,etc).Among the reported cases,female patients predominated with an average male/female ratio of 0.92 ∶ 1.The incidence of syphilis was highest in the age group of 20-39 years,but increased yearly by more than 30％ in the population aged ＞ 60 years.Of 20 occupations,peasant workers accounted for the highest proportion (31.56％) of syphilis cases,and retired population showed the most rapid increase by about 27.35％ per year.The constituent ratio of primary and secondary syphilis in all the syphilis cases had declined yearly,while that of latent syphilis increased by more than 50％ after 2010.Conclusions Syphilis remains a major public health issue in China,and measures based on the epidemiological features are urgently needed to effectively control epidemic syphilis.

Objective To investigate the influence of follow-up treatment compliance on prognosis after gastroesophageal varices treated under endoscopy.Methods Up to 416 liver cirrhosis patients after gastroesophageal varices treated under endoscopy were follow-up and divided into compliance group and control group according to whether the patients had follow-up treatment compliance condition.The factors caused the difference of follow-up treatment compliance were analyzed.The differences in the follow-up indexes such as rebleeding,mortality,rehospitalization were compared between the two groups.The differences in indexes between two groups were stratifiedly analyzed according to liver function Child-Pugh classification.Chi-square four data table test was for two independent samples comparison.Results The differences of education level,income and the medical condition of residential area determined the difference of follow-up compliance.After gastroesophageal varices patients treated under endoscopy,the total rebleeding rate was 35.1 ％ (146/416) and and the mortality rate was 9.4％ (39/416).The rebleeding rate and mortality rate of the compliance group were significantly lower than those of control group (26.1％ (61/234) vs 46.7％(85/182),x2=19.137,P＜0.01; 6.4％(15/234) vs 13.2％(24/182),x2=5.533,P=0.019).Among the 273 Child-Pugh A level patients,the detection rate of liver cancer (3.7％,10/273),liver transplantation rate (3.7％,10/273),splenectomy plus portal azygous disconnection rate (6.6％,18/ 273) of compliance group were higher than those of control group (x2 =4.086,P =0.043; x2 =4.086,P=0.043; x2 =5.515,P=0.019).Among the 102 Child-Pugh B level patients,there were statistical differences between compliance group and control group in rebleeding rate (x2 =21.297,P＜0.01),motality (x2=3.525,P=0.042),ascites (x2=4.451,P=0.035),life quality (x2 =10.454,P=0.001) and liver function (x2 =8.197,P=0.004).However,there were no statistical differences in all indexes between the two groups of Child-Pugh C level patients (all P＞0.05).Conclusion To improve the follow-up treatment compliance remarkably,decreased the rebleeding rate and mortality,contributed to early detection of liver cancer and early liver transplatation,thus the prognosis was improved consequently.

AIM: To prepare urokinase-loaded immunoliposomes, with anti-D-dimer mouse monoclonal antibody that can selectively target to thrombotic site and test the thrombolytic effect in the rabbit model of acute pulmonary embolism (PE) in early phase. METHODS: 40 New Zealand white rabbits were randomly divided into five groups: TBS group (TBS buffer, negative control group), urokinase (UK) group (150 000 IU/kg UK, positive control group), Lip group (Lip-UK, containing 50 000 IU/kg UK), Ab group (Ab-Lip-UK, containing 50 000 IU/kg UK) and 2 Ab group (2 Ab-Lip-UK, containing 50 000 IU/kg UK but double DDmAb of Ab group). PE model was established. Five minutes later, five solutions (TBS, UK, Lip-UK, Ab-Lip-UK, 2 Ab-Lip-UK) were transfused through femoral vein respectively. Right ventricular systolic pressure (RVSP) and right ventricular diastolic pressure (RVDP) were estimated within one hour. The pathological changes of lung, heart, liver and kidney were all observed. RESULTS: RVSP in TBS group had no significant changes within one hour after administration. However, RVSP had respective notable decline at 30 min, 40 min, 30 min, 20 min in UK, Lip, Ab and 2 Ab group, respectively. The means of residual emboli were as follows: 4.0±0 in TBS group, 2.4±0.9 in UK group, 3.1±0.6 in Lip group, 2.4±0.9 in Ab group and 1.9±0.6 in 2 Ab group. The lungs in each group showed scattering local congestion, effusion and swelling on the surface. Obvious hemorrhage of heart, kidney and liver was found with HE staining in UK group, no pathologic change was observed in other groups. CONCLUSION: 2 Ab-Lip-UK with early thrombolytic effect and security may be an ideal thrombolytic agent for treating pulmonary embolism.

Objective To explore the influence of home synthesize magnetic iron oxide (called Fe2O3-PLL) labeling on peripheral blood endothelial progenitor cells (EPCs) bionomics to provide experimental foundation for MR imaging ex and in vivo. Methods Fe2O3 was incubated with PLL for 2 hours to obtain a complex of Fe2O3-PLL. Rabbit peripheral blood mononuclear cells were isolated and EPCs were selected by adherence method. Fe2O3-PLL was used to label EPCs. Prussian blue stain and electron microscope was used for showing intracellular iron. MTT assay was assessed to evaluate the difference of growth curve between unlabeled and labeled with 25 mg/L Fe2O3-PLL. Flow cytometry was performed to analyze cell cycle, cell apoptosis and the expression of surface markers of labeled and unlabeled cells. Expressions of Enos, KDR and Vwf at Mrna levels among unlabeled and labeled EPCs were detected by real-time polymerase chain reaction. Calcium ion channel and membrane fluidity were observed and analyzed by laser confocal microscopy. Statistical analyses were used with ANOVA and t test. Results Almost 100% cells were labeled by Fe2O3-PLL, iron-containing vesicles were intracytoplasma. There was no statistical difference in cells growth curve, cell life cycle [(93.74±3.52)% ,(94.57±3.66)% ] and cell apoptosis rate(12. 89±1.81) %, (11.67±1.18) %) between labeling with Fe2O3-PLL at a concentration of 25 mg/L and unlabeled cells (t = 0. 283, P > O. 05 ; t = 0. 977, P > 0. 05). There was also no statistical difference in relative amount of Enos, KDR and Vwf at Mrna levels and the expression of sudace phenotypic markers (CD34, CD106, CD146 and KDR) between two groups (P > 0. 05). In addition,Labeling had little influence on calcium ion channel and didn't significantly alter cell membrane fluidity.Conclusions The rabbit peripberal blood EPCs can be effective labeled with Fe2O3-PLL and without significant influence on cells bionomics at a low concentration of 25 mg/L. Almost every cell can be labeled and the labeled cells can be used further.