BACKGROUND:As the sequencing technology has been widely used and high-resolution confirmation of organ transplant matching has been gradualy developed, new human leukocyte antigen (HLA) aleles are emerging. However, the gene frequency of some genes cannot be calculated accurately, and there are rare reports. These genes are often ignored, and it is easy to misjudge their genotypes only according to gene frequency. OBJECTIVE:To test and analyze a rare alele, HLA-C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. METHODS: Genomic DNA was extracted automaticaly from the blood sample by using quick DNA purified kit and amplified by HLA-C locus commercial sequence-based typing kit. The purified PCR product was utilized as the DNA template in the sequencing reaction, and six direct sequencing reactions of PCR product covering exons 2, 3 and 4 in both directions were performed using commercial kit. Four direct sequencing reactions of PCR product covering exon 5 in both directions, exon 6 in forward direction and exon 7 in reverse direction were performed using in-house BigDye terminator cycle sequencing reaction kit. Sequencing reaction products purified by ethanol/sodium acetate/ ethylenediaminetetraacetic acid method were sequenced by ABI PrismTM3730 DNA Sequencer. RESULTS AND CONCLUSION:The alele assignment was analyzed with Assign-SBT 3.6+ software, and the sample HLA-C typing result was C*07:04, 08:99. Increasing the sequencing analysis at exons 5, 6 and 7 of HLA-C locus wil help to make clear the ambiguous SBT result and improve the accuracy of HLA-C typing when it is necessary, which shows important significance in clinical tissue matching. Cite this article:Wang DM, Zou HY, Nie DM, Gao SQ, Wang F.Sequencing analysis of a rare human leukocyte antigen, C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):102-106.

Purpose　To inquire into the clinical and pathological features of subcutaneous panniculitis-like T-cell lymphoma. Methods　The histopathology and immunophenotypes of subcutaneous panniculitis-like T-cell lymphoma in 8 cases were observed. Results　Clinically one or more subcutaneous nodules were first found, then the lesion infiltrated up to the epiderm or down to the fascia. Half of the cases were stable for long time without systematic symptoms. The others got high fever with bone marrow, lymphonode involvement, had tumescent liver, and spleen, and decreasing of the counts of thrombocytes or whole blood cells. A few were accompanied with lethal hematophage syndrome. Some rapidly advanced and died less than 3 years. Pathologically, atypical large, midium-size and small lymphocytes with variable ratio rounded the lipocytes like rosettes and were confluent to small patchs or foci or formed a reticular pattern. There were many karyokineses and karyorrhexis made of distinct karyorrhexis bodies. The histocytes reactively proliferated accompanied with phagoerythrocytic phenomena, and polynuclear giant cells and granulomatoid reaction were found. The tumor cells infiltrated in the lipolubules. They expressed T-cell immunophenotype. Conclusions　This tumor is a special type of skin lymphoma involving subcutaneous fat tissue and special clinicophathological features.

Objective:To explore the causes of the maintenance of smoking behaviors in male patient with chronic schizophrenia.Methods:Totally 188 male,smoking patients with chronic schizophrenia,meeting with the criteria of Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition (DSM-IV),above 5-year course of disease,taken a steady dosage of antipsychotics for 3 months,and 61 healthy smoking men were collected according to the voluntary principles.The psychopathologic symptoms and the dependence of cigarette of all patients were as-
sessed with the Positive and Negative Syndrome Scale (PANSS)and the Fagerstrom Test for Nicotine Dependence (FTND),the Russell's Smoking Motivation Questionnaire (SMQ)was used to investigate the causes for maintai-ning behaviors of smoking in patients and healthy controls respectively.Results:The sedative item scores of SMQ were lower in patient group than in the controls [3 (0,9)vs.6 (0,9),P <0.001],and other scores of subscales had no significant difference compared with control.Heavy dependence happened in patient group slightly less than health controls (2.7%vs.8.2%,P <0.05).The total scores of FTND were positively correlated with the doses of antipsychotics (chlorpromazine equivalent)(r =0.22,P <0.01).The scores of total (β=0.22),automatic (β=0.27)and sedative (β=0.37)items in patient group were less than controls (Ps <0.001)by multi-factors step-wise Linear regression analysis.Conclusion:Compared with general smokers,lower need for sedation,less heavy dependence and motivation in smoking patients might be correlated with the strict management for inpatients smok-ing,social-psychological factors and culture background in China.

Objective To further prove the multipotentiality of rabbit bone mesenchymal stem cells (MSCs) by inducing it into cardiomyocytes with 5-azacytidine. Methods MSCs from rabbit were abstracted, isolated, cultured. Then MSCs were induced by 5-azacytidine in vitro for 24 hours. After cultured for 4 weeks, MSCs’ differentiation was studied by immunohistochemistry and electron microscopy technique. Results Some of MSCs induced by 5-azacytidine expressed troponinT . Electron microscopy showed myofilaments. Conclusion MSCs which exist in bone marrow are mutipotential stem cells, and they can be induced into cardiomyocytes by 5-azacytidine in vitro.

AIM: To investigate the expression of heat shock protein 60(HSP60) and Toll-like receptor 4 transduction system in mouse cardiac transplantation. METHODS: The mouse cervical heart transplantation model was established. The animals were divided into control group (the donor and recipient were all C57BL/6 mice) and experimental group (the donor was BALB/c mice and recipient was C57BL/6 mice). The heart and blood were collected for study at 3 d and 7 d. The pathological analysis of the hearts was performed. The levels of cytokines in the serum were determined using ELISA. The expression of HSP60, TLR4, MyD88 and NF-κB in the myocardium was determined by immunohistochemistry and Western blotting. RESULTS: The expression levels of HSP60, TLR4, MyD88 and NF-κB were higher in experimental group than those in control group. Severe rejection was observed in experimental group, whereas no distinct rejection in control group was found. The cytokines (TNF-α, IFN-γ, IL-12) increased significantly in experimental group as compared to those in control group. CONCLUSION: HSP60 increases significantly after heart transplantation, which may activate Toll-like receptor 4 transduction system in a MyD88-dependent pathway and promote allograft rejection. Regulation of HSP60 signal transduction may be a novel way for treating allograft rejection.

BACKGROUND: It was uncertain that the migration, differentiation and survival of transplanted bone marrow mesenchymal stem cells (BMSCs) into myocardium after the acute myocardial infarction. OBJECTIVE: To investigate the migration, differentiation and survival of rabbit transplanted autologous BMSCs in myocardium after the acute myocardial infarction. METHODS: Rabbit BMSCs were isolated and labeled by DAPI in vitro. Rabbit left anterior descending branch was ligated to establish acute myocardial infarction models. Following successful model establishment, 30 New Zealand rabbits were assigned to BMSC and control groups (n = 15). In the BMSC group, autologous BMSCs were infused into the surrounding sites of the infracted region by 4 points 1 hour following coronary artery ligation. In the control group, the same region was injected with an equal volume of saline. Injection volume was 30 μL in each point. Five animals from each group were sacrificed 10 minutes, 3 days and 4 weeks following transplantation. The heart was obtained to undergo frozen sections. The distribution of DAPI-labeled BMSCs was observed using fluorescence microscope. Immunofluorescence method was used to examine the troponin Ⅰ and α-actin. RESULTS AND CONCLUSION: DAPI-labeled BMSCs with blue nuclei were distributed extensively in the myocardium of the cell transplantation group, ovoid in shape and arranged in parallel with the cardiac muscle fibers. Troponin Ⅰ and α-actin were positive immunofluorescently in the cytoplasm of the labeled BMSCs. Results indicated that transplanted BMSCs in the ischemic myocardium could differentiate into myocardial cells under stimulation of local microenvironment.

Objective To explore the clinical value of prospective electrocardiography-triggering high-pitch spiral acquisition by dual-source CT (HP mode ) in children with congenital heart diseases (CHD).MethodsThirty-six patients ( mean age:0.67 years,range:1 month to 2 years and 6 months; 25 males;mean weight:7.7 kg)underwent prospective ECG-triggering high-pitch spiral acquisition.The dose length product (DLP) was recorded to calculate effective dose ( ED ).Two experienced radiologists made diagnosis and assessed the overall image quality by a five-point scale independently.The consistency of their judgment was determined by Kappa statistics.Surgeries were performed in 29 patients,and conventional cardiac angiography(CCA) was performed in 7 patients.The accuracy,sensitivity and specificity of HP mode were calculated based on the surgical or CCA findings.ResultsThe DLP was (5.12 ± 1.64) mGy.cm with a range of 2.4 mGy · cm to 9.2 mGy · cm and ED was (0.125 ± 0.027 ) mSv ranging from 0.078 mSv to 0.179 mSv.The average subjective image quality score was 4.2 ± 0.7.All images were diagnosable.Two radiologists made good agreement ( K =0.774,P ＜ 0.05 ).The accuracy,sensitivity and specificity of HP mode were 98.9％,94.2％ and 99.8％,respectively.ConclusionLow radiation dose with the accurate diagnosis of anomalies makes HP mode a new choice for children with CHD.

Objective: To evaluate effects of color matching of different cavosurface margins on the resin composites in vitro.Methods:Twenty extracted human premolars with an A 2 shade buccal surface were used in this study .Rectangular shaped cavities (3.0 mm depth, 2.0 mm width, 2.0 mm length) were prepared in the center of the buccal surfaces .The gingival and occlusal cavosurface margins were prepared to be either shoulder or bevel;the other cavosurface margins remained vertical .Ten teeth were filled with Clearfil AP-X (AP), the other ten with Clearfil Majesty (MJ) and light cured.The color difference at the cavosurface margin area was measured using a spectrophotometer ( CrystalEye ) and evaluated by 3 observers subjectively .The data were statistically analyzed using repeated measures ANOVA and Chi-square test .Results:When measured by CrystalEye , the color difference between the tooth and resin composite was reduced from the center of restoration to the cavosurface margin area .Both objective and subjective evaluations showed that for AP , the color difference at the cavosurface margin area had no statistical difference among 3 types of the margins; for MJ, the color difference at bevel margin area was significantly smaller than that at the vertical margin area .Conclusion: The resin com-posite restorations produced the color matching at marginal area .The color matching of resin composites with higher diffused light transmission property is more susceptible to the type of cavosurface margins . Preparing bevels may reduce the color difference between the restoration and tooth surface .

Chronic alcohol dependence is an addictive disorder,which is characterized by the excessive craving and uncontrollable drinking.It seriously endangers the patients' psychological and physical health.Recently,MRI has been widely used in the research of substance addiction,especially the alcohol dependence and has made a great progress.It has a great impact on the early prevention,diagnosis,treatment and prognosis of the disease.The progresses of structural and functional MRI in chronic alcohol dependence were reviewed in this article.

Objective To investigate the incidence and risk factors of the complications in perioperative intra-aortic balloon pump (IABP) supported cardiac surgical patients. Methods The clinical data of adult cardiac surgery patients undergoing IABP in Fuwai Hospital from January 2005 to January 2017 were enrolled. The patients were divided into complications group and no complications group. Demographic characteristics, diagnosis, perioperative clinical parameters, IABP related data, and IABP complications (including ischemia, bleeding, vascular injury and mechanical problems) were collected. The incremental risk factors of complications related IABP were analyzed by logistic regression. Results During the 12-year period, 522 patients received IABP support, with 388 male and 134 female; the mean age was (61.79±9.35) years; the complications related to IABP occurred in 25 patients, and overall complication rate was 4.79%; 87 IABP patients were dead in-hospital, the overall mortality was 16.67%, no patient died due to complications. The complications rate was higher in the female patients (40.00% vs. 24.95%), and was more in patients with age ≥ 65 years old (80.00% vs. 38.03%), more with higher body mass index [BMI (kg/m2): 25.45±13.71 vs. 22.95±3.45], diabetes mellitus (44.00% vs. 26.76%), combination treatment with extra-corporeal membranous oxygenation (ECMO: 20.00% vs. 5.03%) and prolonged IABP support time (hours: 134.4±90.3 vs. 109.8±89.1, all P < 0.05). There was no significant difference in the incidence of complications among preoperative IABP support, intra-operative IABP support and post-operative IABP support [3.30% (3/91), 5.46% (10/183), 4.84% (12/248), χ 2 =0.629, P = 0.730]. Bleeding from puncture site occurred in 14 cases (2.68%) without severe bleeding. Limb ischemia occurred in 9 cases (1.72%). One patient (0.19%) was under another surgery because of retroperitoneal hemorrhage caused by vascular injury. One patient (0.19%) was unsuccessful due to a balloon leak. It was shown by logistic regression analysis that presence of age ≥ 65 years [odds ratio (OR) = 2.320, 95% confidence interval (95%CI) = 1.011-1.806, P = 0.047], diabetes mellitus (OR = 2.281, 95%CI = 1.016-5.120, P = 0.026) and combination treatment with ECMO (OR = 4.341, 95%CI = 1.240-15.196, P = 0.040) were found to be the risk factors of complications related to IABP. Conclusions IABP complication rates are generally low. The frequent complications during IABP support is bleeding from site of catheterization and limb ischemia. When patients were treated with IABP, those with older age, diabetes mellitus and combination with ECMO should be monitored closely in order to reduce complications.