Purpose:To study the effects of hydroxycamptothecin with thermotherapy on anti-angiogenesis in vitro and in vivo. Methods:We chose human microvascular endothelial cell (HMVEC) culture and chick embryo choriallantoic membrane (CAM) model and used MTT and number-calculating methods to observe hydroxycamptothecine on HMVEC’s proliferation , sprouts and CAM blood vessels’ formation.Results:The survival rate of endothelial cells was in the range of (68.2%)-44.7% within the dose of 20-40 ng/ml and 5-80 ng/ml and was negatively correlated with the concentration (correlation coefficient was -0.906,-0.469,P=0.00003,0.0051). Hydroxycamptothecine could significantly suppress the endothelial cells’ proliferation and the sprouts. Hydroxycamptothecine could significantly suppress CAM vessels. The survival rate of HepG II cells is in the range 100%-90% within the dose of 5-80 ng/ml. There was no cytotoxicity.There was a synergestic anti-angiogenetic effect when hydroxycamptothecin (20 ng/ml) was combined with thermotherapy in vitro while there was additive effect when hydroxycamptothecin (40 ng/ml) was combined with thermotherapy in vitro.Conclusions:This experiment shows that small doses of hydroxycamptothecine (20-40 ng/ml) with thermotherapy has anti-angiogenetic synergestic or additive effect on proliferation and migration both in vivo and in vitro.

Objective To evaluate the comparability of the detection results of different blood cells analysis systems in same hos-pital.Methods Referring to the Guideline for Comparability Verification of Quantitative Test Results in Medical Institutions,the comparability validation protocol was established.The EDTA-K2 anticoagulation fresh whole blood samples with proper concentra-tion were detected for 5 parameters of HGB,RBC,HCT,PLT and WBC by 4 systems (Sysmex XT-1800i,Sysmex XT-2000,Sys-mex XT-4000i and Mindray BC-5800).The range was calculated and the detection results consistency analysis was performed.Re-sults The acceptable standard of critical differentials was intended to be HGB3.5%,RBC3%,HCT3%,PLT12.5% and WBC 7.5%.The replication detection was at least 2 times and up to 5 times.The ranges of 3 concentrations after replication detection and sample comparison were 2.87%-6.29%,1.57%-2.99%,1.95%-4.77%,12.81%-25.74% and 6.72%-11.13% respective-ly.The ranges of RBC detection results in 4 systems were smaller than the critical differentials,the validation was passed.The ran-ges of HGB,HCT,PLT and WBC detection results in 4 systems all had the condition of more than the critical differentials,the vali-dation did not passed.After removing the test system with obvious bias,the validation of the detection results by other test systems was passed.Conclusion The RBC detection results by 4 systems have the comparability;the HGB,HCT and PLT detection results by Sysmex XT-1800i,Sysmex XT-2000i and Sysmex XT-4000i have the comparability;the WBC detection results by Sysmex XT-1800i,Sysmex XT-2000i and Mindray BC-5800 have the comparability.

Objective To establish a real-time quantitative reverse-transcription PCR ( qRT-PCR) method for detection of hepatitis E virus ( HEV) of different genotypes based on standard HEV DNA plasmid in order to promote its application in clinical laboratory. Methods Specific primers and probe of HEV were designed based on the conserved open reading frame 3 (ORF3) regions. HEV DNA plasmids were construc-ted and 10-fold serial dilutions of the plasmids were prepared and used as standards to establish one-step qRT-PCR. The established method was compared with HEV antigen, antibody and RT-nPCR assays. Some positive samples were sequenced and analyzed by evolutionary tree. Results The one-step qRT-PCR meth-od for HEV detection in serum or feces samples was successfully establish. It could reach a sensitivity of 25 copies/test and 77. 8% of its results were consistent with those by HEV antigen assay. Nine patients were infected with HEV of genotypes 4a, 4d or 4n as indicated by evolutionary tree. Conclusion The HEV qRT-PCR method based on its standard plasmid is successfully established, which paves the way for commercial-ization of clinical applications.

BACKGROUND:As an important cytokine, interleukin-6 regulates immune responses in inflammation sites and has an autocrine/paracrine activity that stimulates osteoclast formation and bone resorption, which is related to bone remodeling during orthodontic tooth movement.
OBJECTIVE:To investigate the effects of orthodontic force on the expression of interleukin-6 mRNA in the periodontal tissue of rats.
METHODS:In situ hybridization was performed to measure the expression of interleukin-6 mRNA at 1, 3, 5, 7, 10 and 14 days after the application of orthodontic force on the maxil ary first molars of rats.
RESULTS AND CONCLUSION:The expression of interleukin-6 mRNA was observed at a low level in the normal periodontal tissue of rats. After the application of force, the induction of interleukin-6 mRNA was observed to reach a maximum on day 3 and to decline thereafter. The expression of interleukin-6 mRNA can be evoked by orthodontic force but with a certain self-limiting. As a multifunctional cytokine, interleukin-6 plays a very important role in periodontal remodeling during orthodontic tooth movement.

Objective To analyze the precision and accuracy of the Olympus AU5421 automatic biochemical analyzer in order to verify the performance of this detection system declared by the manufacturer.Methods The Precision and Accuracy of User Au-thentication-Guide for Approval Second Edition(CLSI EP15-A2)was used to perform the routine detection on the Olympus AU5421 automatic biochemical analyzer.The systematic precision and accuracy were analyzed.Results Under this experiment condition,the precision and accuracy of the Olympus AU5421 automatic biochemical analyzer was consistent with the performance declared by the manufacturer.Conclusion The Olympus AU5421 automatic biochemical analyzer system has the high precision and good accuracy, and can be better applied in the clinical routine detection.

Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.

Adult ; Female ; Hemoptysis ; etiology ; Humans ; Pulmonary Artery ; abnormalities ; Recurrence ; Tomography, X-Ray Computed

1/2 cycle, three cases

Objective To estimate the consistency between the diagnostic criteria for dermatomyositis (DM) and polymyositis (PM) developed by Bohan and Peter criteria (B/P criteria) and ENMC criteria.Methods The clinical,laboratory and pathological data from 86 patients who were initially diagnosed with idiopathic inflammatory myopathy were collected retrospectively.These patients were diagnosed according to B/P criteria and ENMC criteria,and the similarities and differences between these two criteria were compared.The data were analyzed with Mann Whitney U test and Kappa test by SPSS 13.0 software.Results Thirtyseven DM and 49 PM were diagnosed using B/P criteria.Forty-six DM and 14 PM were diagnosed using ENMC criteria,and 1 was diagnosed as eosinophilic myositis,9 were diagnosed as sporadic inclusion body myositis (sIBM),11 cases were diagnosed as limb-girdle muscular dystrophy type 2B,and the diagnosis of 5 patients could not be clarified.Agreement for DM between these two sets of criteria was very good by Kappa test (κ=0.79),but the corresponding between the two tests for PM was poor (κ=0.26).Conclusion Our study has demonstrated that B/P criteria may cause over-diagnosis and misdiagnosing for PM.ENMC criteria involves immunohistochemical pathology,stratified clinical and pathological exclusion criteria.The diagnostic accuracy of ENMC criteria is much improved.

Background Keratectasia after laser in situ keratomileusis (LASIK) is a rare but severe complication,which threatens the visual acuity and corneal strength.Corneal collagen crosslinking (CXL) is a new therapy that increases the security and decreases the risk of complication.However,the effectiveness and safety of LASIK-CXL is still need to be concerned.Objective This study was to evaluate the safety of LASIK-CXL for myopia and astigmatism with thin cornea.Methods A prospective cohort study was designed.A total of 128 eyes of 64 patients with thin corneal and myopic astigmatism enrolled in Beijing Tongren Eye Center from January 2014 to January 2015.The patients were assigned to LASIK group (74 eyes of 37 patients) and LASIK-CXL group (54 eyes of 27 patients).Refractive surgery was performed by Visumax femtosecond lasrer and VISX S4 excimer laser.Eyes of LASIK-CXL group applied accelerated CXL immediately after LASIK.The follow-up was 6 months.Manifest refraction,uncorrected (UDVA) and corrected distance visual acuity (CDVA),average keratometry values (AveK),anterior segment OCT (AS-OCT),corneal hysteresis (CH) and corneal resistance factor (CRF) were examined before and after operation.This research passed through Ethics Committee of Beijing Tongren Hospital.Results The spherical equivalent (SE) of the LASIK group and LASIK-CXL group were (-6.49 ±2.41)D and (-6.97 ±2.41) D before operation and decreased to (-0.68 ±0.88) D and (-0.75 ±0.94) D 6 months after operation.The UDVA (LogMAR) was 1.18±0.28 and 1.05±0.38 before operation and elevated to-0.06±0.09 and-0.03±0.186 months after operation in the LASIK group and LASIK-CXL group.The preoperative AveK values were (44.37 ±1.46) D and (44.47± 1.50)D in the LASIK group and LASIK-CXL group and reduced to postoperative (39.30±2.06) D and (38.66± 1.80) D.The preoperative SRI of LASIK group and LASIK-CXL group were 0.25 ±0.21 and 0.24±0.22,which increased to 0.29±0.24 and 0.28±0.24.The SAI values were 0.36±0.16 and 0.39±0.15 before operation,which increased to 0.57 ±0.31 and 0.75 ±0.376 months after operation,and the SAI value of the LASIK-CXL group was significantly higher than that of LASIK (F =10.220,P--0.002).CRF values of LASIK and LASIK-CXL were (8.44±1.44)mmHg and (8.63±1.35) mmHg in preoperation,which decreased to (5.74±1.31) mmHg and (6.25± 1.24) mmHg in postoperation.The result of LASIK-CXL was higher than that of LASIK (F=8.650,P =0.040).CH values were 8.78 ± 1.51 and 8.69 ± 1.62 in preoperation,which decreased to (7.23 ± 1.08) mmHg and (6.50±1.32)mmHg.The value of LASIK-CXL was lower than that of LASIK (F =5.860,P =0.017).The mean depth of demarcation line was (228.45±28.24) μm (range 165 to 310 μm) on OCT,which was presented in 45 eyes (81.82％) at 1 month in postoperation.Conclusions Accelerated CXL with FS-LASIK is effective and safe in improving visual acuity in myopic astigmatism patients with thin cornea,which also can increase the rigidity of the cornea.