Objective To determine the effect of nanochemotherapy drug on Survivin and p53 ex-pressed by human biliary traet carcinoma cell line QBC939.Methods Culturing the human biliary tract carcinoma cell line QBC939 in vitro and it was divided into five groups including saline,nanochemotherapy drug,nanopartiele withoul nanochemotherapy drug,5-FU and gemcitabine.Using the methods of MTT and flow cytometry to observe the growth of QBC939 which was dealt with different drugs.In addition,RT-PCR and Western blot were used to delect the expression of mRNA and protein by Survivin or p53.Results The expression of mRNA and protein by Survivin decreased in the following set:saline,nanoparlicle withoul nanochemotherapy drug,5-FU,gemcitabine and nanochemotherapy drug,respeclively.However,the ex-pression of mRNA and protein by p53 were in reverse order.The inhibiting action to QBC939 was obvious in nanochenmtherapy drng and the apoplotic rate was higher than others except for gemcitabine(P＜0.05). Conclusion Nanochemotherapy drug has significant effect on treatment cholangiocarcinoma in vilro,which may attribute to the down regulation of Survivin and up regulation of p53.

Objective To detect plasma amino-terminal pro-brain natriuretic peptide (NT-ProBNP) ,D-Dimer levels in the pa-tients with acute pulmonary embolism (APE) in order to investigate their change characteristics and significance .Methods Among 60 patients with suspected APE ,40 cases diagnosed by CT and meeting the thrombolytic condition were set as the experimental group and other 20 cases of excluded APE by CT as the control group .Plasma NT-ProBNP and D-dimer before treatment in the two groups and after 2-week thrombolytic therapy in the experimental thrombolysis were detected and compared .Results The diagnos-tic sensitivity of NT-ProBNP for early APE was 92 .3% and the specificity was 65% ,while the diagnostic sensitivity of D-dimer for early APE was 100% and the specificity was 70% ;the plasma NT-ProBNP and D-dimer levels before thrombolysis in the experi-mental group were significantly higher than those in the control group with statistical differences (P<0 .01) .Conclusion Plasma NT-ProBNP and D-dimer has important clinical significance for APE and can provide the basis for the early diagnosis and the cura-tive effect observation of the patients with APE .

Objective To evaluate the value of ultrasound (US) in assessment knee joint inflammation in patients with juvenile rheumatoid arthritis(JRA).Methods US scans of the knees obtained in 30 children at clinically active stage; JRA was compared with those obtained in 30 healthy children and 10 JRA patients in clinical remission.Results Changes in synovial membrane thickness and presence of fluid in suprapatellar bursa showed statistically significant differences between JRA patients with active disease and the other subjects.Alterations in contour of the articular cartilage were demonstrated in 10 knees of patients with JRA.Conclusion US is a simple sensitive and reliable methods for the assessment and monitoring of knee joint involvement in JRA.

Species identification of biological samples is widely used in such fields as forensic science and food industry. A variety of accurate and reliable methods have been developed in recent years. The current review shows common target genes and screening criteria suitable for species identification, and described various DNA-based molecular biology methods about species identification. Additionally, it discusses the future development of species identification combined with real-time PCR and sequencing technologies.
Animals ; DNA/genetics* ; Genetic Techniques/trends* ; Humans ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA/trends*

BACKGROUNDObstetric hysterectomy (OH) as a lifesaving measure to manage uncontrolled uterine hemorrhage appears to be increasing recently. The objective of this study was to determine the etiology and changing trends of OH and to identify those at particular risk of OH to enhance the early involvement of multidisciplinary intensive care.

METHODSA retrospective study was carried out in patients who had OH in China-Japan Friendship Hospital from 2004 to 2014. Maternal characteristics, preoperative evaluation, operative reports, and prenatal outcomes were studied in detail.

RESULTSThere were 19 cases of OH among a total of 18,838 deliveries. Comparing the study periods between 2004-2010 and 2011-2014, OH increased from 0.8/1000 (10/12,890) to 1.5/1000 (9/5948). Indications for OH have changed significantly during this study period with uterine atony decreasing from 50.0% (5/10) to 11.1% (1/9) (P < 0.05), and placenta accreta as the indication for OH has increased significantly from 20.0% (2/10) to 77.8% (7/9) (P < 0.05). Ultrasonography and magnetic resonance imaging (MRI) have been used to make an exact antepartum diagnosis of placenta accreta. A multidisciplinary management led to improved outcomes for patients with placenta accreta.

CONCLUSIONAs the multiple cesarean delivery rates have risen, there has been a dramatic increase in OH for placenta accreta. An advance antenatal diagnosis of ultrasonography, and MRI, and a multidisciplinary teamwork can maximize patients' safety and outcome.

Adult ; Female ; Humans ; Hysterectomy ; trends ; Patient Care Team ; Placenta Accreta ; surgery ; Pregnancy ; Retrospective Studies

Objective To establish a HRM assay to screen for KRAS mutations in clinical colorectal cancer patients.Methods The sensitivity of HRM was analyzed by detecting somatic mutations in exon 2,notably codons 12 and 13 of the KRAS gene in the serial plasmid mixture samples which were mixed using the different proportions mutation plasmid and wide type plasmid of KRAS.HRM analysis was performed for KRAS on DNA insolated from a panel of 60 colorectal cancer samples derived from fresh tissues.The results were compared with the direct sequencing data.Results After the PCR amplification,the mutation results could be available by performing HRM analysis in the same tube on a real time PCR machine with HRM capability.HRM detection could identify KRAS mutation in a proportion of 10% of mutation plasmid DNA.All 60 samples identified the KRAS mutation by HRM and sequencing.17 samples were positive(28.3%) by HRM for KRAS exon 2 mutations,and 15 samples were confirmed the presence of codon 12 or 13 mutations(25.0%) and the other 2 samples were wild type by sequencing.The 60 samples detected by HRM were given 100% sensitivity with 96% specificity.Conclusions HRM is a sensitive intube methodology to screen for mutations in clinical samples.HRM will enable high-throughput screening to gene mutations to allow appropriate therapeutic choices for patients and accelerate research aimed at identifying novel mutations in human cancer.

Objective According to international classification criteria (2002) on Sjogren' s syndrome, labial pathology was still considered as a major criterion for diagnosis. Standard labial biopsy was hard to be carried out in China. This study is to evaluate whether the invasive labial biopsy could be replaced by noninvasive detection of serum anti-SSA antibody. Methods 181 Chinese patients with the initial diagnosis of primary Sjogren's syndrome in Peking Union Medical College Hospital (PUMCH) were enrolled in Sjogren's International Collaborative Clinical Alliance (SICCA). All patients received standard labial biopsies (area of salivary gland tissues≥4 mm~2) and focal score (FS) of focal lymphatic sialadenitis were confirmed by pathologists from school of stomatology,University California of San Francisco (UCSF). Anti-SSA antibodies in sera of all patients were detected by double immunodiffusion (DID), Western blot in PUMCH and by enzyme-linked immunosorbent assay (EIJSA) in central laboratory of SICCA. The correlation between labial pathological findings and serum anti-SSA antibody was studied by X~2 test and the concordance was calculated by unweighted Kappa. Results(1)Bivariate analysis revealed strong associations of FS > 1 with the presence of anti-SSA antibody by DID (83.9% vs 42. 0%, P <0. 0001). The accordance between FS and antibody detection by DID was fine with a kappa value of 0. 432. However, there were 16. 1% false-positive antibody reports and 42.0% false-negative antibody reports. (2)FS > 1 was strongly associated with the presence of anti-SSA antibody by Western blot (83.0% vs 51.7%, P < O. 0001). But the accordance between FS and antibody detection by Western blot was only fair with a kappa value of 0. 316. There were 17.0% false-positive antibody reports and 51.7% false-negative antibody reports. (3)FS > 1 was strongly associated with the presence of anti-SSA antibody by ELISA (81.5% vs 38.6%, P <0. 0001). The accordance between FS and antibody detection by EI,ISA was fine with a kappa value of 0.427. There were 18.5% false-positive antibody reports and 38. 6% false-negative antibody reports. Conclusion In Sjogren's syndrome, labial biopsy with FS > 1 finding is strongly associated with anti-SSA antibody. Positive results of anti-SSA antibodies by DID or ELISA may indicate FS > 1, thus labial biopsy could relatively be avoided, negative results may need further standard labial biopsy procedure to confirm the diagnosis of Sjogren's syndrome.

Objective To summarize the characteristics of common carotid arterial wave intensity (WI) in healthy volunteers and offer the normal reference values of WI. Methods Common carotid arteries (CCA) in 525 healthy volunteers were examined using Aloka Prosound α10 equipped with wave intensity software. The volunteers were divided into six group according to age,group A,＜20 years old;group B,20 - 29 years old;group C,30 - 39 years old;group D,40 - 49 years old;group E,50 - 59 years old and group F,≥60 years old. Carotid arterial wave intensity in normal subjects had two positive peaks, the first peak,W1 ,and the second peak, W2. Between the two positive peaks, a negative area (NA), the time interval between the R-wave of ECG and the first peak(R-W1 ) and that between the first and second peaks (W1-W2) were observed. Results ① 1050 common carotid arteries in 525 volunteers were studied. The mean values of W1,W2,NA,R-W1,and W1-W2 were (8332±4796) mmHg · m-1 · s-3,(1853±1143)mmHg · m-1 · s-3,(38 ± 26) mmHg · m-1 · s-2, (102 ± 19)ms and (266 ± 42)ms,respectively. ②These values were no significant between the left and right CCA in same group. W1 and NA were correlated with age ( r = - 0. 204 and r = -0.301, P ＜0. 001). W1 and NA in group A and B were significant increased than those in other groups( P ＜0. 05 or P ＜0. 001 ). ③There were statistical significances in W1 and W1-W2 of CCA between male and female. ④There were no statistical significances in all values in common carotid arteries of both sides (P ＞0. 05). ConclusionsWI technique is useful for evaluating the dynamic behavior of the heart and the vascular system and their interaction. W1 is very sensitive to the changes in the working condition of the cardiovascular system.

Objective:To express fusion protein of mouse thymic stromal lymphopoietin (TSLP) and HIV-1 gp120BAL V1/V2 subdomain in 293F cell.Methods:Full length of the V1V2 sequence from BAL isolate was fused with the C-terminus of mouse thymic stromal lymphopoietin (TSLP) and sub-cloned into pCEP-Pu vector to construct the eukaryotic expression plasmid-pCTV1V2BAL.The recombinant plasmid was confirmed by enzyme digestion and sequencing , then transfected into 293 F cells using PEI as a transfection reagent .The fusion protein was purified by metal chelate affinity chromatography and characterized by SDS -PAGE and Western blot . The epitopes of V1/V2 in fusion protein were identified by ELISA .Results:The SDS-PAGE and Western blot results showed that there were highly heterogeneous glycoprotein bands at the site between 35 kD and 55 kD, which reacted with anti-mTSLP rabbit polyclonal antibody and anti-His tag mouse monoclonal antibody .The ELISA analysis showed that antibodies to V 1/V2BAL existed in the sera of HIV-1 positive patients.Conclusion:The mTSLP-V1/V2 fusion protein was successfully expressed in 293F cells, which may be useful for HIV-1 vaccine research .

OBJECTIVE To investigate the effect of mono-2-ethylhexyl phthalate(MEHP) on proliferation of primary neural stem cells(NSCs)of rats and NE-4C cells of mice and on the migration of NE-4C cells and the mechanism. METHODS NE-4C or NSCs were treated with MEHP 1,10,100 and 1000 μmol · L-1 for 72 h,respectively. The cytotoxicity was estimated with the cell counting kit-8 (CCK-8). Cell proliferation was analyzed by EdU assay. The mRNA expression levels of the glucocorticoid receptor(GR),signal transducer and activator of transcription 3(Stat3)and sex determining region Y (SRY)-box 2(Sox2) were detected by qRT-PCR. The protein expression levels of total GR,GRβ, Sox2,Stat3 and p-Stat3 were measured by Western blotting. RESULTS Cell viability of NE-4C cells and NSCs at MEHP 1000μmol·L-1 was significantly decreased,which was 70.3%and 40.0%of the control group, respectively. EdU assay showed that MEHP 100 μmol · L-1 decreased NE-4C cells and NSCs by 74.8%and 12.0%(P<0.05)compared with control. The effect of MEHP on the cell migration of NE-4C was evidenced by the fact that the migration was obviously reduced to (63.4±2.0)%(P<0.05)after treatment with MEHP 100μmol · L-1 for 72 h. The mRNA expression levels associated with proliferation and migration in NE-4C of GR,Stat3 and Sox2 in MEHP 100 μmol · L-1 group were down-regulated to 49.8%,26.0% and 14.0%of control(P<0.05). At MEHP 100μmol · L-1,mRNA of GR, Stat3 and Sox2 in NSCs declined to 10.0%,14.0% and 15.3% of normal control. Western blotting results revealed that protein expressions of GR,GRβ,Sox2 and p-Stat3 were remarkably inhibited by MEHP 100 μmol · L-1 in that the relative expression of NE-4C was 0.92 ± 0.17,0.87 ± 0.35,0.81 ± 0.22 and 0.62 ± 0.24(P<0.05). The corresponding protein expression in NSCs was 0.82 ± 0.20,0.56 ± 0.12,0.84 ± 0.36 and 0.53 ± 0.20(P<0.05)when the cells were treated with MEHP 100μmol · L-1 for 72 h. CONCLUSION MEHP can inhibit the proliferation and migration of NE-4C cells and NSCs possibly by decreasing Stat3 and Sox2 that are mediated by GRβ.