Objective To observe the feasibility of 5.0T MR for cardiac imaging.Methods Three patients with heart diseases and 17 healthy volunteers were prospectively enrolled.Cardiac MR(CMR)cine sequence and black blood sequence imaging were performed using 5.0T and 3.0T MR scanner,respectively.The image quality and artifacts degrees were compared between 5.0T and 3.0T CMR images,and the consistency of left ventricular parameters obtained using 5.0T and 3.0T scanners was analyzed.Results No significant difference of image quality nor artifacts degrees was found between 5.0T and 3.0T CMR images(all P＞0.05).The left ventricular end diastolic volume(EDV),end systolic volume(ESV),ejection fraction(EF),stroke volume(SV)and end diastolic mass(EDM)derived from cine images acquired at different fields were in a good agreement(all ICC＞0.75,all P＜0.001).Conclusion 5.0T MR could be used for cardiac imaging,with image quality of cine and black blood sequences comparable to that of 3.0T MR.

Based on the principle of magnetic anastomosis technique, the design of magnetic anastomosis system for endoscopic tissue clamping is proposed. The system includes a semi-ring magnet, a special structure transparent cap and a detachable push rod. With the help of the existing digestive endoscopy and endoscopic tissue gripper, the endoscopic close clamping and anastomosis of the bleeding or perforated tissue can be completed. After the anastomosis, the magnet falls off and is discharged through the digestive tract. Animal experiments showed that the system was easy to use, the fistula was clamped firmly, the magnet was discharged for 7~21 days, and there was no magnet retention and digestive tract obstruction. Further safety verification, optimization of endoscopic operation, the system can be used in clinical trial.
Anastomosis, Surgical ; Animals ; Constriction ; Endoscopy, Gastrointestinal ; Magnetics ; Magnets

Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.

Objective To analyze the 10-year survival outcome and failure patterns for patients with nasopharyngeal carcinoma (NPC) after intensity-modulated radiotherapy (IMRT),aiming to provide reference for optimized treatment for NPC.Methods Clinical data of 866 patients with NPC receiving IMRT from January 2001 to December 2008 were retrospectively analyzed.Survival analysis was performed using the Kaplan-Meier estimator.Univariate analysis was carried out by log-rank test and multivariate analysis was performed using Cox proportional hazards model.Results The median follow-up time was 132 months.The 10-year local recurrence-free survival (LRFS),distant metastasis-free survival (DMFS),progression-free survival (PFS) and disease specific survival (DSS) were 92.0％,83.4％,75.7％ and 78.6％,respectively.A total of 210 patients died including 124 patients (59.0％) from distant metastasis,which was the primary cause of death,and 47 (22.3％) from local regional recurrence.Independent negative factors of DSS included age＞50 years (P=0.00),LDH ≥ 245 IU/L (P=0.00),Hb＜ 120 g/L (P=0.01),T2-T4 staging (P=0.00),N1-N3 staging (P=0.00) and GTV-nx＞20 cm3(P=0.00).The 10-year LRFS,DMFS and DSS of stage Ⅱ NPC patients did not significantly differ after IMRT alone and chemoradiotherapy (P=0.83,0.22,0.23).For patients with stage Ⅲ NPC,the 10-year LRFS and DSS in the chemoradiotherapy arm were significantly higher than those in the IMRT alone (P=0.01,0.01),whereas no statistical significance was observed in the DMFS between two groups (P=0.14).The overall survival of stage Ⅳa+Ⅳb NPC patients is relatively poor.Conclusions IMRT can improve the long-term survival of NPC patients.Distant metastasis is the primary failure pattern.Patients with stage Ⅰ-Ⅱ NPC can obtain satisfactory survival outcomes after IMRT alone.The addition of chemotherapy can further enhance the LRFS and DSS of stage Ⅲ NPC patients.However,the optimal therapeutic strategy remains to be urgently investigated for stage a+ Ⅳb NPC patients.

OBJECTIVE: To establish a cell lines for quality control of prenatal genetic diagnosis. METHODS: The recombined SV40LTag-pcDNA3.1(-) vector was constructed and transfected by lipidosome into human amniotic fluid cells with common aneuploidy. Positive clones were screened by G418, and the immortality of transfected cell line was identified. RESULTS: Cell line with karyotype of 46, XY, t(8;19)(q24.3;q13.1) from primary amniotic fluid cells was established. Karyotype analytical results indicated that the cell line at its 15th generation maintained the same karyotype of its primary cell. CONCLUSIONS Gene can lead to immortality of amniotic fluid cells, which contributes to preparing cell lines for internal and external quality control in prenatal genetic diagnosis.
Antigens, Polyomavirus Transforming ; genetics ; Cell Line ; Female ; Genetic Vectors ; Humans ; Karyotype ; Pregnancy ; Prenatal Diagnosis ; methods ; Quality Control ; Recombinant Proteins ; genetics ; Transfection

Objective To explore the value of serum microRNA-155-5p and -133a-3p (miR-155-5p and miR-133a-3p) expression for the diagnosis and prognosis evaluation of sepsis. Methods A prospective observational study was conducted. 105 sepsis patients admitted to emergency intensive care unit (EICU) of the First Affiliated Hospital of Zhengzhou University from January 2015 to January 2016 were enrolled. They were divided into three groups according to the severity: 35 patients with sepsis, 35 with severe sepsis, and 35 with septic shock. At the same time, 35 healthy persons were selected as the control group. According to the prognosis, the patients were divided into improved group (n = 70) and in-hospital death group (n = 35). The clinical data of all the subjects were collected. The mRNA expressions of miR-155-5p and miR-133a-3p were determined by reverse transcription-polymerase chain reaction (RT-PCR). The receiver-operating characteristic curve (ROC) was plotted to evaluate their clinical value for the diagnosis and prognosis of sepsis. The binary logistic regression was used to analyze the risk factors affecting the prognosis of sepsis patients. Results ① The mRNA expressions of serum miR-155-5p and miR-133a-3p were gradually increased with the aggravation of sepsis. The mRNA expression of miR-155-5p (2-ΔCt) in sepsis, severe sepsis, sepsis shock groups was 1.89±0.48, 2.21±0.41, 2.79±0.73 (F = 23.737, P = 0.000), and the mRNA expression of miR-133a-3p (2-ΔCt) was 1.38±0.31, 1.74±0.65, 2.08±0.47, respectively (F = 27.710, P = 0.000). It was shown by ROC curve analysis that the area under the ROC curve (AUC) of serum miR-155-5p and miR-133a-3p for the diagnosis of sepsis was 0.855 [95% confidence interval (95%CI) = 0.761-0.949] and 0.769 (95%CI = 0.666-0.872) respectively. The cut-off value of miR-155-5p for the diagnosis of sepsis was 1.64, the sensitivity was 85.3%, and specificity was 80.6%. While the cut-off value of miR-133a-3p was 0.82, the sensitivity and specificity were 97.9% and 54.8% respectively. ② Compared with improved group, the patients of in-hospital death group were more serious, and procalcitonin (PCT), C-reactive protein (CRP), D-dimer, lactic acid (Lac), sequential organ failure assessment (SOFA) score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, and the mRNA expressions of miR-155-5p and miR-133a-3p were significantly increased (all P < 0.05). While there was no statistically significant difference in gender, age, white blood cells (WBC), serum creatinine (SCr) between the two groups (all P > 0.05). It was shown by binary logistic regression analysis that Lac [odds ratio (OR) = 0.514, 95%CI = 0.260-0.893, P = 0.024], sepsis severity (OR = 0.039, 95%CI = 0.023-2.955, P = 0.016), SOFA score (OR = 0.668, 95%CI = 0.474-0.825, P = 0.001), serum miR-155-5p expression (OR = 0.117, 95%CI = 0.020-0.530, P = 0.007) were the risk factors affecting the prognosis of patients with sepsis. Conclusions The expression of serum miR-155-5p and miR-133a-3p may be used as specific indicators for the diagnosis of sepsis. And the expression of miR-155-5p can be used as independent impact factor for the estimation of sepsis prognosis.

Objective To construct the prokaryontic expression vector of the gene fragment which encodes the transpeptidase domain of penicillin binding protein 2a(PBP2a) of methicillin‐resistant Staphylococcus aureus(MRSA) ,and to express ,purify and i‐dentify the objective protein .Methods Strains of MRSA were isolated and identified from clinical samples ,according to the se‐quence of mecA gene recorded in GenBank ,the primers of mecA fragment which encoded the transpeptidase domain of PBP2a was designed .The gene fragment from MRSA was amplified by using polymerase chain reaction(PCR) and cloned into pET28a(+ ) plasmid .After being identified by enzyme digestion and sequencing ,the recombinant plasmid was transformed into the strain of Escherichia coli BL21(DE3)plysS .The expression of transpeptidase domain of PBP2a was induced by 0 .7 mmol/L IPTG ,the ex‐pressed products were purified by using Ni afinity chromatography ,then were analyzed by using Western blot .Results The recom‐binant expression vector was digested by BamHⅠ and EcoRⅠ ,and the products were at the expected size .The result of sequencing showed two bases undergoing mutation ,while there were no frameshift mutations .The expressed protein was identified by using SDS‐PAGE and Western blot ,a new protein band was visible at the relative molecular mass of 38 × 103 .Conclusion The corre‐sponding prokaryotic expression vector is successfully constructed ,and the transpeptidase domain of PBP2a is successfully ex‐pressed and purified .

Objective To preliminarily evaluate the performance of glucose detention reagents with three kinds of different chro-mogens and to investigate their anti-interference performance according to NCCLS document.Methods According to the protocol EP10-A2 provided by the National Committee for Clinical Laboratory Standards(NCCLS),the samples with low,middle and high level of glucose were detected by the glucose reagent kits with 3 kinds of different chromogens.The bias,total imprecision,inter-cepts,slope rates,nonlinearities,carryover rate and drifts were calculated.The interference evaluation test was performed according to the document EP7-A2.Results The bias and total imprecision of three kinds of reagent kits were all within allowed ranges.No statistically significant differences were showed in intercepts,slope rates,nonlinearities,carryover rate and drifts.1450 turbidity chyle,5 g/L hematoglobin and 0.03 g/L vitamin C did not interfere with the assay of three kinds of glucose reagent kits with differ-ent chromogens.342 μmol/L free bilirubin,342 μmol/L conjugated bilirubin did not interfere with the detection of reagent with MAOS.Conclusion The glucose detention reagents with three different chromogens have good accuracy and precision,and various performance indexes all conform to the clinical application requirements,reagent with chromogen MAOS is better than other chro-mogenic reagents in the anti-interference performance.

Objective To improve EP9-A2 for Bias estimation among multiple quantitative detection systems within full range of AMR.Methods 40 patients specimens were determined twice for serum total cholesterol by four detection systems(A,B,C and D).With system A served as comparative method,Bias between A and the other three was evaluated according to CLSI EP9-A2 separately.Furthermore,DD(distance from deviation to tolerable error)and its average confidence intervals between every two sys-tem were calculated and compared with zero.The confidence interval of greater than zero was served as criteria for accepting bias between every two system.Results Bias between A and the other three meet the analytical quality specification according to EP9-A2,although that of D system was positive,and those of B and C system were negative.DD between every two system obeyed nor-mality distribution.All biases between every two system wes acceptable except that between B and D,causing of their interval con-taining zero.After correcting of results from system D,Biases between every two system were all acceptable.Plots of confidence in-terval could provide a full range bias assessment within AMR.Conclusion Comparability and Bias estimation in full range of AMR for results between every two system among 3 or more systems could be evaluated by confidence intervals.

Myosin heavy chain 9 (MYH9)disorders are a group of ihherited thrombocytopenias resulted from the mutation of MYH9 gene,including May-Hegglin anomaly,Epstein syndrome,Fechtner syndrome and Sebastian syndrome.MYH9 disorders are very often misdiagnosed as idiopathic thrombocytopenic purpura (ITP).For better understanding of MYH9 of clinical and laboratory and getting enough attention in clinical practice,this review will focus on the pathogenesis,clinical manifestations,laboratory examination and differential diagnosis.