Optimized serological isolation of lung-cancer-associated antigens from a yeast surface-expressed cDNA library

The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.     

Percutaneous Radio-Frequency Rhizotomy in the Treatment of Trigeminal Neuralgia

A percutaneous technique of selective partial trigeminal root coagulation was evaluated in the treatment of 38 patients suffering from trigeminal neuralgia, 1 patient with pain secondary to oral carcinoma and 1 patient with atypical facial pain. The pain of trigeminal neuralgia was relieved in 94.7 percent of patients. Pain was relieved in the patient with oral carcinoma, but not in the patient with atypical facial pain. There was no mortality and no permanent morbidity outside of the trigeminal nerve lesion. The procedure requires only a brief hospital stay without the time, expense and hazards of open cranial surgical procedures.     

The best strategy of blood transfusion in patients with Rhnull syndrome

Rh_null syndrome is a very rare disease. Patients with this syndrome present with negative serological Rh typing of E, e, C, c, and D antigens. Only one study has previously discussed Rh_null syndrome in Chinese individuals. We experienced two patients with Rh_null syndrome in China, Rh genotypes being CcDEe in the first patient and CCDee in the second patient. The first patient was a pregnant woman (gravida 2, para 1) with a negative red blood cell (RBC) antibody screen test. The second patient was a middle-aged man, transfused with ccdee, ccdEe, and ccdee RBC products, the pre-transfusion specimen was negative and post-transfusion specimen was anti-c,e, respectively. The hemoglobin level continued to increase in the second patient after being transfused with ccdEe RBC products. In the first patient, the result of the antibody screen test was still negative after artificial abortion. In patients with Rh_null syndrome, RBC products that have the same Rh genotype as the patient can be safely transfused.     

Events upstream of mitochondrial protein import limit the oxidative capacity of fibroblasts in multiple mitochondrial disease

To investigate whether protein import is defective in mitochondrial disease, we compared the rate of import and the expression of protein import machinery components in skin fibroblasts from control subjects and a patient with multiple mitochondrial disease (MMD). The patient exhibited a 35% decrease in cytochrome c oxidase activity and a 59% decrease in cellular oxygen consumption compared to control. Western blot analyses revealed that patient levels of MDH, mtHSP70, HSP60, and Tom20 protein were 57%, 20%, 75% and 100% of control cells, respectively. MDH and Tom20 mRNA levels were not different from control levels, whereas mtHSP70 mRNA were 50% greater than control. Radiolabeled MDH was imported into mitochondria with equal efficiency between patient (44% of total synthesized) and control (43%) cells, although the total MDH synthesized in patient cells was reduced by about 40%. The unaffected levels of mRNA and post-translational import into mitochondria, combined with reduced protein levels of MDH, mtHSP70, and HSP60 suggest a translational defect in this patient with MMD. This was verified by the 50% reduction in overall cellular protein synthesis in the patient compared to control. Further, the similar import rates between patient and control cells suggest an important role for Tom20, but a lesser role for mtHSP70 in regulating protein import into mitochondria.     

Regional mapping of clotting factors VII and X to 13q34. Expression of factor VII through chromosome 8

Summary Blood clotting factors were investigated in a patient with trisomy 8 mosaicism, a patient with an r(13), and a patient with distal trisomy 13q. Results are compatible with the assignment of the structural genes of factors VII and X to 13q34 and the existence of a regulatory mechanism associated with chromosome 8 controlling the expression of factor VII alone.     

Refractoriness at peripheral and pituitary receptors in general and pituitary types of thyroid hormone resistance.

Patients with the general type (patient #1 and #2) and the selective pituitary type (#3) of thyroid hormone refractoriness (TR) were studied to clarify defects at peripheral and pituitary receptors. Products of T3 and TSH (n = 63) were calculated when T3 was above the normal limit (T3 > 1.8 ng/ml, 2.8 nmol/l) as one of the indices of pituitary resistance. Means of T3 (ng/ml) x TSH (mU/l) of patient #1 (mean; 40.8), #2 (15.0) and #3 (8.6) were significantly greater than patients with Graves' disease (2.1), suggesting pituitary refractoriness in the 3 patients. The products of patient #1 and #2 were also significantly larger than patient #3, demonstrating that the pituitary insensitivity in the latter (#3) was less than the former patients. Means of serum cholesterol in patients #1 and #2 were higher than patient #3 and patients with Graves' disease. Products of T3 (ng/ml) and cholesterol (mg/ml) (n = 28) in the patient #1 (541.9) and #2 (461.0) were significantly greater than the patient #3 (292.8) and the patients with Graves' disease (275.3). The results demonstrate generalized refractoriness in the patient #1 and #2 and selective pituitary resistance in the patient #3. It is suggested that our patient with the pituitary type (#3) had less severely affected receptors at the pituitary than our two patients with the general type. These results are consistent with the previous hypothesis that the pituitary type of TR is a partial form of this disease.     

肥胖患者HFA小鼠模型的建立

目的研究肥胖患者的肠道菌群在无菌小鼠体内的定植规律。方法选取20只无菌KM小鼠,接种肥胖患者的粪便,构建菌群人源化(HFA)动物模型,利用变性梯度凝胶电泳技术(DGGE)评价患者肠道菌群在无菌小鼠体内的定植规律。结果 HFA小鼠菌群平均丰富度(richness,S)为12.04±3.68,肥胖患者的条带S为24,为HFA小鼠S的2倍;肥胖患者Shannon指数(H')为3.02,HFA小鼠平均H'为2.46±0.33;HFA小鼠与人肠道菌群的总相似度为26%;大部分雌性HFA小鼠与雄性HFA小鼠在聚类分析图上分离,且雄性HFA小鼠与患者更为相似。结论HFA小鼠体内能部分模拟肥胖患者的微生物区系,且与患者性别相同的小鼠模拟得更好。本实验建立的HFA模型为肥胖与肠道菌群关系的进一步研究提供新的选择。     

Determination of antithrombocyte antibodies in the blood serum of patients with idiopathic thrombocytopenic purpura by an immunoenzyme method

Enzyme-linked immunosorbent assay (ELISA) was developed for determination of serum antiplatelet antibodies. Platelets obtained from healthy donors of blood group 0(1) were washed off plasma and sedimented on the bottom of microtest wells. After washing off unattached platelets and blocking of plastic with albumin platelets were incubated with sera under investigation and binding of serum antibodies was detected using antihuman immunoglobulin antibodies conjugated with peroxidase. Ten patients with idiopathic thrombocytopenic purpura (ITP). 1 patient with systemic lupus erythematosus. 1 patient with red blood cell aplasia and 9 healthy donors (negative control) were studied by ELISA. Serum antibodies which effectively bound to platelets were detected in 5 patients with ITP, in patient with lupus erythematosus and in patient with red blood cell aplasia.     

Control of blood-transmitted infections in dentistry

As knowledge has accumulated on the blood-transmitted pathogenic agents, the contact with biological fluids (blood, plasma, saliva, etc.) from apparently healthy individuals has started to be regarded as a real professional risk for dentists. Theoretically, exposure to a contaminated biological specimen may have as a consequence transmission of infection from patient to dentist, from dentist to patient and from patient to patient via inadequately decontaminated and sterilized dental equipment. The present study is concerned with the analysis of the specific conditions that favor the occurrence of the epidemic process, the estimation of the risk degree of transmission of infections caused by hepatitis B, C viruses as well as of HIV infection in Romania. The data for the study were collected using two processes. First a self reporting survey and secondly an experimental procedure were performed. The testing of dentists' knowledge of blood transmissible diseases and infection control in their offices were performed using a questionnaire with 129 questions. The professional incidents/accidents representing a potential risk were counted using a questionnaire (with 37 questions). Serological markers were tested with ELISA kits. The monitoring of sterilization was accomplished with a questionnaire and biological tests. Many conclusions result from the study. There is an extremely reduced probability and infection transmission from the dentist to the patient. The transmission of infection from the patient to the dentist represents a low risk (for all that, the risk should not be minimized). The rigorous control and observation of infection prevention measures in dental offices is necessary to stop the infection transmission from patient to patient. The dentists' postgraduate training in infection control measures should be completed with knowledge regarding the blood transmissible infections epidemiology. Learning more about the epidemiological process enables the dentists to avoid wrong attitudes and behaviors.     

A Case Study of a Patient with Gliosarcoma with an Extended Survival and Spinal Cord Metastases

Gliosarcoma is a rare brain tumor consisting of both glial and mesenchymal components. Metastatic gliosarcoma is rare; however, here we report a 31-year-old Chinese woman with cranial gliosarcoma metastatic to the liver, lymph nodes and the spinal cord. Initially, the patient presented with dizziness, headache and vomiting and after surgery and histological examination, was diagnosed with cranial gliosarcoma. The patient was treated with surgical resection followed by chemotherapy and radiotherapy. Three years after completing treatment, the patient again presented with similar symptoms with the addition of a seizure. Test revealed recurrence of the gliosarcoma, and the same treatment was prescribed. Three years after treatment completion, the patient again presented with dizziness and headache. Masses at the right temple and in the right side of the neck were found. Tumors were surgically removed from the brain, skull, scalp and neck, the latter three diagnosed as metastatic gliosarcomas. The patient received both chemotherapy and radiotherapy following resection. One month after treatment, bone scans revealed possible metastasis in the right skull, lumbar and left ileum, soft neck tissue, lungs, collarbone, humeri, vertebrae, liver and abdominal lymph nodes. No further therapy was recommended due to the poor condition of the patient. The patient died 5 months later.     

Axillary lymphadenopathy: a complication of BCG immunotherapy for melanoma

We present a patient with a Clark level IV melanoma who received BCG immunotherapy by scarification for 12 months after wide excision. Six months after cessation of treatment, the patient developed regional lymphadenopathy that histology showed to be reactive epithelioid granuloma. Regional immunotherapy thus represents a potentially confusing factor in the post-operative assessment and management of the patient with melanoma.     

Hemorrhagic Shock As the Initial Manifestation of Pheochromocytoma: Report of A Sequential Management Strategy

ObjectiveTo describe a patient presenting with hemorrhagic shock attributable to bleeding pheochromocytomas and the sequential management strategy used for treating this patient.MethodsWe summarize the clinical presentation,diagnostic work-up, surgical management, and pathologicfeatures of our patient and review the pertinent literature.ResultsA 38-year-old man with multiple endocrineneoplasia type 2A and bilateral pheochromocytomas presented initially with nearly fatal retroperitoneal and intraperitoneal hemorrhage rather than the characteristic hypertensive paroxysms. After lifesaving operative interventionand a 5-month period of rehabilitation and convalescence,the patient underwent bilateral retroperitoneoscopic adrenalectomy as definitive treatment. Thus, the abdomen thathad been operated on multiple times because of hemorrhage was left undisturbed, and the patient had a successfulrecovery.ConclusionNear-fatalintraperitonealhemorrhageis a very rare initial manifestation of pheochromocytoma.Our current patient with bilateral pheochromocytomaspresented in this dramatic manner. This case shows that a sequential management strategy of damage-control surgical treatment followed by future resection of the tumorsafter appropriate a-adrenergic blockade is a safe and effective therapeutic option. (Endocr Pract. 2012;18:e81-e84)     

Continuity of care in general practice: effect on patient satisfaction.

OBJECTIVE--To evaluate the influence of continuity of care on patient satisfaction with consultations. DESIGN--Direct and episodic specific evaluation of patient satisfaction with recent consultation. SETTING AND SUBJECTS--A representative sample of 3918 Norwegian primary care patients were asked to evaluate their consultations by filling in a questionnaire. The response rate was 78%. MAIN OUTCOME MEASURES--The patient''s overall satisfaction with the consultation was rated on a six point scale. Continuity of care was recorded as the duration and intensity of the present patient-doctor relationship and as patients'' perception of the present doctor being their personal doctor or not. RESULTS--The multivariate analysis indicated that an overall personal patient-doctor relationship increased the odds of the patient being satisfied with the consultation sevenfold (95% confidence interval 4.9 to 9.9) as compared with consultations where no such relationships existed. The duration of the patient-doctor relationship had a weak but significant association with patient satisfaction, while the intensity of contacts showed no such association. CONCLUSION--Personal, continuous care is linked with patient satisfaction. If patient satisfaction is accepted as an integral part of quality health care, reinforcing personal care may be one way of increasing this quality.     

Application of orthogonal light scattering for routine screening of lymphocyte samples

Orthogonal and forward light-scattering properties of lymphocytes were measured from patients with different lymphocytic diseases in order to determine the potential value of light scattering as a screening device. Monitoring of orthogonal light scattering of lymphocytes of a B-cell chronic lymphocytic leukemia patient during splenic irradiation (SI) revealed the selective decrease of malignant cells and the fact that the major part of the residual lymphocytes were cytotoxic lymphocytes. By combining forward and orthogonal light scattering it was shown that lymphocytes from a patient with T gamma lymphocytosis were abnormal. Orthogonal light scattering also showed an increase in cytotoxic lymphocytes in a patient with mononucleosis infectiosa and in a splenectomized patient. Orthogonal light scattering of lymphocyte subpopulations showed that the leu8+ population of a patient with mononucleosis infectiosa was bidisperse. For elderly donors the occurrence of CD3+, CD4+, CD8+, and HNK-1+ lymphocytes with a large orthogonal light scattering varied considerably. The CD8+ lymphocytes of these donors consisted mainly of cytotoxic lymphocytes. These results show that determination of light-scattering properties of lymphocytes may yield important diagnostic information and can indicate when further investigation of the lymphocytes by means of immunofluorescence is necessary.     

PEX13 is mutated in complementation group 13 of the peroxisome-biogenesis disorders.

The peroxisome-biogenesis disorders (PBDs) are a genetically and phenotypically diverse group of diseases caused by defects in peroxisome assembly. One of the milder clinical variants within the PBDs is neonatal adrenoleukodystrophy (NALD), a disease that is usually associated with partial defects in the import of peroxisomal matrix proteins that carry the type 1 or type 2 peroxisomal targeting signals. Here, we characterize the sole representative of complementation group 13 of the PBDs, a patient with NALD (patient PBD222). Skin fibroblasts from patient PBD222 display defects in the import of multiple peroxisomal matrix proteins. However, residual matrix-protein import can be detected in cells from patient PBD222, consistent with the relatively mild phenotypes of the patient. PEX13 encodes a peroxisomal membrane protein with a cytoplasmically exposed SH3 domain, and we find that expression of human PEX13 restores peroxisomal matrix-protein import in cells from patient PBD222. Furthermore, these cells are homozygous for a missense mutation at a conserved position in the PEX13 SH3 domain. This mutation attenuated the activity of human PEX13, and an analogous mutation in yeast PEX13 also reduced its activity. The mutation was absent in >100 control alleles, indicating that it is not a common polymorphism. Previous studies have demonstrated extragenic suppression in the PBDs, but the phenotypes of patient PBD222 cells could not be rescued by expression of any other human PEX genes. Taken together, these results provide strong evidence that mutations in PEX13 are responsible for disease in patient PBD222 and, by extension, in complementation group 13 of the PBDs.     

Improvement of anemia by recombinant erythropoietin in patients with myelodysplastic syndromes and aplastic anemia

Eight patients with myelodysplastic syndromes (MDS) and four patients with aplastic anemia (AA) were treated with recombinant erythropoietin (rEpo) to investigate its effect on the anemia of these patients. rEpo was administered by i.v. injection three times a week for at least four weeks. The doses were 3,000, 6,000, or 12,000 U/day. Despite an elevated "endogenous" Epo level, a greater than 1.5 g/dl increase in hemoglobin (Hb) concentration was observed in one patient with refractory anemia (RA), one patient with refractory anemia with excess of blasts (RAEB), and one patient with AA. A greater than 50% decrease in red cell transfusion requirement was observed in one patient with RA and one patient with AA. One RA patient and one AA patient have received rEpo as maintenance therapy for more than 64 and 100 weeks, respectively. They no longer need red cell transfusions and have had a normal Hb concentration and normal ferrokinetics. No side effect was seen. These results indicate that rEpo may benefit some patients with MDS and AA who are dependent on red cell transfusions while further studies will be necessary to elucidate the mechanism by which rEpo stimulates erythropoiesis and improves anemia in patients with these diseases.     

Immunophenotypic analysis of simultaneous specimens from different sites from the same patient with malignant lymphoma

Immunophenotypic analysis of simultaneous specimens from different sites from the same patient with malignant lymphoma The assumption that immunophenotypic characteristics of different specimens obtained simultaneously from the same patient remain unchanged has rarely been evaluated. Using flow cytometry, we reviewed our experience of 29 patients with non Hodgkin's lymphoma (NHL). From these patients, 60 simultaneous specimens taken from the peripheral blood, bone marrow, effusions, fine needle aspirates from lymph nodes or cerebrospinal fluid were studied. In 26 out of 29 patients, the immunophenotype in the different specimens was identical. In one patient with unclassifiable low-grade B-NHL, immunophenotyping showed additionally a CD38 expression in the effusion which was not seen in the bone marrow. In one patient with mantle cell lymphoma, expression of CD10 in the lymph node was noted which was lacking in the peripheral blood. In the remaining patient with unclassifiable low-grade B-NHL, CD23 expression was noted in the lymph node but not in the peripheral blood. This retrospective study suggests that discordant antigen expression in samples from different body sites within the same patient is a rare event.     

原发性肝癌合并下腔静脉癌栓的手术治疗

目的：探讨原发性肝癌合并下腔静脉癌栓的手术治疗方法。方法：采用肝切除加下腔静脉取栓治疗2例肝癌合并下腔静脉癌栓患者,取栓方法包括经荷栓肝静脉取栓（1例）和下腔静脉切开取栓（1例）,又分在全肝血流阻断下取栓和在萨氏钳局部血管阻断下取栓。结果：2例肝癌及下腔静脉癌栓均得到成功切除,术中无明显并发症发生;术后无死亡;随访中一例存活11月;另1例已生存8个月。结论：肝癌合并下腔静脉癌栓的手术治疗安全可行,其基本术式为肝切除加下腔静脉切开取栓。     

服务蓝图在出入院服务流程优化中的应用及效果分析

目的 对出入院服务流程进行梳理与优化，提高服务效率、改善患者体验。方法 利用服务蓝图确定住院患者的出入院服务体验、内部服务流程、后台支持保障系统等，并对服务效率和患者体验进行评价。结果 在某三甲综合性医院建立出入院服务中心，统筹协调所有出入院相关服务后，患者术前待床时间、办理出入院手续时间、术前检查等待时间和往返距离，均比去年同期有所降低。结论 将服务蓝图应用于医院出入院服务流程的优化，通过资源合理配置、跨部门协作、信息平台支撑、规章制度保障，能够有效地保障患者安全、提高运行效率、提高患者满意度。