Objective To evaluate the endocrine glands derived VEGF (EG-VEGF) influence on growth, migration and apoptosis of pancreatic cancer cells MiaPaCa Ⅱ. Methods MiaPaCa Ⅱ were treated by 100,200 ng/ml EG-VEGF for 24, 48, 72, 96 h, and MTT assay was used to determine the proliferation; and cell scratch experiment was used to investigate the percentage of cell migration distance, flow cytometry was used to measure the apoptosis of the cancer cells. Results After MiaPaCa Ⅱ cells were treated by 0, 100,200 ng/ml EG-VEGF for 72 h, the proliferation of MiaPaCa Ⅱ was 0. 253 ± 0. 012 , 0. 374 ± 0.013,0. 383 ±0.015, respectively EG-VEGF could significantly promote the proliferation of MiaPaCa Ⅱ ( P ＜ 0. 05 ). After MiaPaCa Ⅱ cells were treated by 0, 100 ng/ml EG-VEGF for 24 h, the percentage of cell migration distance was (27.40 ± 3.45 ) % and ( 13.21 ±4.65 ) % ,respectively with statistical difference ( P ＜ 0.05 ), EG-VEGF could significantly promote the migration ability of MiaPaCa Ⅱ cells and inhibite the apoptosis. Conclusions After EG-VEGF treatment, the growth and migration ability of MiaPaCa Ⅱ cells increases, apoptosis decreases.

Objective To investigate the anti-apoptosis effects of EG-VEGF on pancreatic cancer cell MiaPaCa and its molecular mechanism. Methods The cells were treated with 50, 100, 200 ng/ml EG-VEGF. Flow cytometry was used to determine the apoptosis. The expression of p42/44MAPK, STAT3 protein and the phosphorylation, and anti-apoptosis protein Mcl-1 was evaluated by Western blot. Non-specific G protein-coupled receptor antagonist PTX, Rsa/ERK signal transduction blockade PD98059, JAK/STAT3 signal transduction blockade AG490 were used to treat the cells for 1 h, and the change of Mcl-1 protein was observed. Results After treated with 50 ng/ml EG-VEGF, the apoptosis rate of MiaPaCa was decreased from (28.4 ±4.6)% to (13.21 ±4.65)% (P<0.05) ; the phosphorylation of p42/44MAPK increased by 1.735 ± 0.019 folds; the phosphorylation of STAT3 increased by 21.810 ± 0.052 folds; the expression of Mcl-1 protein increased by 3.460 ±0.002 folds when compared with that of control group,and the difference was statistically significant (P < 0.05 ). But the degree of phosphorylation and the expression of Mcl-1 were not further increased with 100, 200 ng/ml EG VEGF treatment. After PTX pre-treatment, the increase of Mcl-1 protein expression was completely inhibited, and after PD98059, AG490 pre-treatment, the increase of Mcl-1 expression was inhibited to 52% and 68%. Conclusions EG-VEGF can inhibit MiaPaCa cell from apoptosis,and the mechanism may be related with activation of Ras MAPK and JAK STAT3 signal transduction pathway and up-regulation of Mcl-1.

OBJECTIVE:To discuss the influence of drug price reduction and regulation of purchase on the use of anti-infectives in our hospital.METHODS:The use of anti-infectives in5times of drug price reduction was retrospective anal?ysed.RESULTS:The proportion of the sum of money for consumption of anti-infectives in the total sum for drug consumption dropped year by year but the total DDDc of anti-infectives increased year by year.The DDDs and the total DDDc of all kinds of anti-infectives showed different characteristics.CONCLUSION:The macroscopic price reduction for anti-infectives posi?tively affects the rational use of anti-infectives,yet the effect is limited.Microcosmic regulation of purchase offers an indis?pensable effect.

Objective To study the distribution of antinuclear antibodies ( ANAs) in a healthy population and the significance of using ANAs screening test in medical examination .Methods The ANAs were measured by indirect immunofluorescence assay ( IIF) .The Western blot assay was used to detect fif-teen specific antibodies against auto-antigens .Results 3519 out of all 25 110 subjects showed ANAs titers>1∶100 , and among them male and female subjects were respectively accounted for 1143 and 2376 .1489 out of all subjects had ANAs titers >1∶320 , and among them male and female subjects were respectively accounted for 406 and 1083 .The positive rates of ANAs at different titers showed significant differences be -tween male and female subjects .Among subjects with ANAs titers >1∶320 , the number of male subjects showed a steady increase with the age , while the percentage of female subjects reached to two peaks during the periods of puberty and menopause .The fifteen specific antibodies were detected in 659 out of 1489 sub-jects with ANAs titers>1∶320 and anti-Ro-52 (14.2%) accounted for the majority , followed by anti-M2 (12.7%) and anti-SSA (9.6%).Conclusion ANAs can be detected among healthy population of all ages, but their distribution varied with gender and age .ANAs screening test is necessary for medical exami-nation of healthy population , especially for female during period of puberty or menopause .The population with positive ANAs should be followed-up closely and educated for the prevention of autoimmune diseases .

Objective To develope a new Real-time quantitative PCR assay using SYBR green as fluorescence reporter, which is rapid, specific, sensitive, cheap and accurate for the detection of mycoplasma pneumoniae(MP), and evaluated its clinical application value.Methods The sequence of the 23S rRNA gene in MP type strain FH was selected as amplified regions, and specific primers were designed.Then the related plasmids were extracted as standards,and the absolute quantitative standard curve was established.The sensitivity ,specificity of the fluorescence quantitative PCR assay was compared with the nest-PCR and kit;To calculate correlation coefficient, coincidence rate and kappa coefficient, clinical samples were detected using above-mentioned methods and cultivation,respectively.Results The detection sensitivity of the new real-time PCR and nest-PCR was 10 copies of FH DNA,while the kit 100 copies.In the specificity tests,the MP sample was positive,while mycoplasma hominis and other four bacteria were all negative.We applied this real-time PCR assay ,nest-PCR, kit and cultivation to 182 clinical specimens, and the detection rates were 55.49％, 52.75％, 47.25％ and 39.01％ ,respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and nest-PCR were 89.6％ ,0.790, respectively;while those of the new method and cultivation were 83.5 ％ ,0.678, respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and the kit were 89.6％ ,0.792,respectively;and the correlation coefficient of these two methods was 0.923,P ＜ 0.001.Conclusion Compared with other methods, the new real-time PCR assay could be used to detect mycoplasma pneumoniae quickly and economically, with high sensitivity and specificity ,revealing great utility value on varied instrumentation platforms.

Objective:To observe the clinical effect of deep insertion at Tianshu(ST 25)for colonic slow transit constipation(STC).Methods:120 cases of STC patients were randomly divided,60 cases in a deep insertion group,30 cases in an electroacupuncture group and 30 cases in a medication group by 2:1:1 ratio.The deep insertion group was treated with deep insertion at Tianshu(ST 25).The electroacupuncture group was treated with routine insertion at Tianshu(ST 25).The medication group was treated with oral administration of Lactulose oral liquid.The first voluntary defecation time,and constipation scores before the treatment,four weeks after the treatment and relevant scores of clinical symptoms were assessed in the three groups of the patients.Results:The scores of the clinical symptoms in improvement of constipation were better in the deep insertion group than in the electroacupuncture group and medication group,with differences in statistical significance(P<0.01).The unsuccessful numbers in the improvement of defecation and abdominal pain were also better in the deep insertion group than in the other two groups,and better in instant effect in the deep insertion.Conclusion:The improvement of STC clinical symptoms was better by deep insertion at Tianshu(ST 25)than by medication and routine acupuncture method at Tianshu(ST 25).

Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.

Objective To investigate the antimicrobial susceptibility of spectinomycin?minocycline?azithromycin and sparfloxacin to mycoplasma(Uu and Mh)and therapeutic effect of spectinomycin to my-coplasma infection in genitourinary tract.Methods①The susceptibility test:each of the4drugs was divided into two concentrations.One was at1?g/mL(sensitive concentration)and the other was at4?g/mL(resistant concentration).If mycoplasma does not grow in both concentrations,it means the drug tested is sensitive.If it grows in both concentrations,the drug tested is resistant.If mycoplasma grows in lower concentration and does not in higher concentration,it means moderate sensitive.②Treatment regimen:Spectinomycin was injected,2g/d IM,for7-10days as a course of treatmeant.Patients were followed-up7days later and2~4weeks after treatment.Results①Among1658specimens,519were found Uu positive,and61Mh positive.The resis-tance rates of Uu to4different drugs were:7.7%for minocycline,21.4%for sparfloxacin,13.9%for azithromycin and7.3%for spectinomycin.Whereas,those of Mh were:18.0%,45.9%,54.1%,and29.5%re-spectively.②The clinical effect of spectinomycin was:out of43treated patients,37(86.0%)cured,4(9.3%)markedly improved,2(4.7%)failed.Total effective rate was95.3%and so was the elimination rate of my-coplasma.Conclusion The resistant rate of mycoplasma to spectinomycin is lower than that to minocycline?azithromycin and sparfloxacin,and the former is widely used in the treatment of mycoplasma(especially Uu)infection,with a satisfactory clinical effect.

Objective To developed A laboratory diagnosis of Moraxella catarrhalis by an laboratories diagnostic method real-time fluorescence quantitative PCR assay. Methods The specific primers and probes were designed based on the sequence of outer membrane protein CopB(copB)gene in Moraxella catarrhalis,and the Taqman probe RT-PCR method was developed to detect the Moraxella catarrhalis.The standard plasmids ex-tracted from the Moraxella catarrhalis standard strains were used to constitute the standard samples,and compared with these standard samples,the sensitivity of the fluorescence quantitative PCR assay was tested by the estab-lished standard curves.The specificity of the fluorescence quantitative PCR assay was tested by the DNA samples of other bacterias in the laboratory.Meanwhile,321 throat swab samples from inpatient and outpatient child pa-tients,with asthma infection were collected as clinical samples to validate the fluorescence quantitative PCR as-say.Results The standard curve was drawn in the real-time PCR by the Taqman fluorescence reporter.During the sensitivity tests,the newly-developed real-time fluorescence PCR could detect at least 10 copies of Moraxella catarrhalis,and could successfully distinguish several DNAs of the pathogens.On the basis of the validation result of the 321 throat swab samples,there are 25 Moraxella catarrhalis with 7.79 % positive rate.Conclusion The fluorescence quantitative PCR assay is of great sensitivity and specificity,and it can be widely used for the detec-tion of Moraxella catarrhalis.

Objective:To investigate drug resistance gene in Mycoplasma pneumoniae(MP) and the distribution of 13 respiratory pathogens in bronchoalveolar lavage fluid(BALF) of children with Mycoplasma pneumoniae pneumonia(MPP).Methods:A total of 100 BALF of children with MPP in Peking University Third Hospital and Peking University First Hospital from January 2018 to January 2019 were collected.Fluorogenic quantitative PCR was used to detect nucleic acid and it′s drug resistance gene of MP and multiple PCR method was adopted to detect influenza A virus, influenza A virus-H 1N 1, influenza A virus-H 3N 2, influenza B, human parainfluenza virus, adenovirus, human bocavirus, human rhinovirus, Chlamydia pneumoniae, human metapneumovirus, MP, human coronavirus, and respi-ratory syncytial virus gene, and the results were compared by using Chi square test. Results:In 100 BALF samples, MP and drug resistance gene were detected by fluorogenic quantitative PCR.Totally, 83 cases (83.00%) were MP positive and 78 cases (93.98%) were drug resistant.All of them had the point mutations A2063G in V region of 23S rRNA domain.A total of 13 kinds of respiratory pathogens were detected by multiplex PCR method, and 89 cases (89.00%) were positive.Totally, 79 cases (79.00%) were MP positive, of which 74 cases (74.00%) detected only MP, and 5 cases (5.00%) detected MP combined with other pathogens.Other pathogens were detected in 10 cases (10.00%). The virus detection rate of 0-4 years old group was higher than that of >4-6 years old group ( P=0.042) and >6 years old group ( P=0.002), and the differences were statistically significant. Conclusions:MP can be detected in most BALF samples of MPP children, the drug resistance phenomenon is serious, and the main point mutation is A2063G.There were other respiratory pathogens and 2 or 3 pathogens were detected in a small number of BALF samples.