Objective To develop allele specific oligonucleotide(ASO) -PCR assay based on TCR βgene rearrangements and provide a screening method for minimal residual disease (MRD) in adult patients with T-lineage acute lymphoblastic leukemia (T-ALL).Methods DNA samples from newly diagnosed 20 adult T-ALL patients were obtained.The TCR β gene rearrangements were detected by multiplex PCR,which included 38 paired of primers in 3 reaction tubes.Gel electrophoresis and two-color Gene Scanning was also applied for clonality analysis of TCR β followed by sequencing and subsequent blasting for monoclonal PCR products in four patients.ASO primers were designed based on the sequence of junction regions.MRD were detected in the bone marrow by RQ-PCR with ASO upstream primers, consensus Jβprobes and downstream primers.Results The detection rate of the clonal TCR β rearrangements was 85.0% (17/20).At least one complete Vβ-Jβ rearrangement could be detected at the time of diagnosis in 16 out of 17 patients(94.1%, 16/17).Incomplete Dβ-Jβ rearrangement could be detected in 7 patients (41.2% ,7/17).The positivitity rate of Vβ-Jβ to Dβ-Jβ was 2∶1 (94.1% versus 41.2% ).Two-color Gene Scanning analysis showed the Jβ2 family was used more frequently than the Jβ1 family (73% versus 27% ).The slopes of the standard curves ranged from - 3.60 to - 3.27.The correlation coefficients of all four standard curves were more than 0.99.The detection sensitivity of ASO-PCR was 4 × 10 -5 μg/μl.The fluorescence background were detected at a low level.Quantitative MRD values of TCR β rearrangement in sequential BM specimens of 4 adult T-ALL patients were monitored during the treatment, including complete remission after induction and after consolidation therapy. RQ-PCR showed the MRD values of TCR β rearrangement were gradually decreased in response to the treatment.Conclusions The quantification of TCR β rearrangement by ASO-PCR approach is sensitive, specific and reliable for the accurate evaluation of malignant clones.It is suitable for the monitoring of minimal residual disease of adult T-ALL patients.

Objective To investigate the application of colostomy care products in managing the leakage around PTCD drainage tube, and to discuss its clinical effect. Methods A total of 56 patients with malignant obstructive jaundice, who had received PTCD and suffered from postoperative leakage around PTCD drainage tube, were randomly and equally divided into the study group (n=28) and the control group (n=28). The colostomy care powder, the skin protective film and the colostomy bag were used for the patients of the study group, while iodophors, sterile gauze and mupirocin ointment were employed for the patients of the control group. The incidence of irritant dermatitis around PTCD drainage tube, the degree of comfort to the indwelling tube judged by the patient, and the nursing workload for PTCD drainage tube were determined, and the results were compared between the two groups. Results The incidence of irritant dermatitis around PTCD drainage tube in the study group was obviously lower than that in the control group. And the degree of comfort to the indwelling tube judged by the patient in the study group was much higher than that in the control group. The differences between the two groups were statistically significant (P<0.05). Conclusion Combination use of colostomy care powder, skin protective film and colostomy bag can effectively reduce the incidence of irritant dermatitis around PTCD drainage tube and improve the patient’s condition. Therefore, this method is worthy of popularization in clinical practice.

OBJECTIVETo analyze the drug use in our burn ward, and to investigate the methods in controlling the proportion of drug expenses, as well as the rational use of antibiotics.

METHODSA randomized sample of 290 burn-patients with hospital stay more than 5 days in 2005 were enrolled in the study, and information about their drug use, (especially that of antibiotics), the drug expenses and its proportion in the total medical expenses, the total therapeutic expenses, and the healing rate of the patients were statically analyzed.

RESULTSSulfamido was dominant among the topically applied antibiotics, while cephalosporins was dominant among the systemic applied antibiotics. The drug expenses accounted for (11 +/- 5)% in the therapy expenses, while the antibiotics expenses accounted for (5.8 +/- 1.7)% in the therapy expenses, but it accounted for (51 +/- 17)% in the drug expenses. The average expense of every patient was 22026.09 RMB, and the healing rate of the patients was 96.90%.

CONCLUSIONThe proportion of drug expense (especially that of antibiotics), as well as total medical expenses of burn patients can be lowered through a combined therapy.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; economics ; therapeutic use ; Burns ; drug therapy ; economics ; Child ; Child, Preschool ; Drug Costs ; Drug Utilization ; statistics & numerical data ; Female ; Humans ; Infant ; Male ; Middle Aged ; Young Adult

Objective To evaluate the anti-infection effects of biodegradable implant for sustained release of cefuroxime,which can be implanted into the anterior chamber in rabbits. Methods The implants for sustained release of cefuroxime were prepared with appropriate mixtures of cefuroxime axetil(CAE),poly(lactic-glycolic) acid(PLGA) and polyvinyl pyrrolidone by solvent evaporation method.Fifty rabbits were selected and divided into experiment group(n=35) and control group(n=15).The concentrations of cefuroxime in aqueous humor 0.5,1,2,6 and 24 h after subconjunctival injection of 125 mg cefuroxime were measured in control group,and those in aqueous humor and plasma 1,2,3,5,7,14 and 28 d after implantation into anterior chamber were detected in experiment group.Besides,for experiment group,the intraocular pressure before and after the implantation were obtained;the inflammation of anterior chamber was routinely observed by slit lamp;and cornea tissues were harvested for scanning electron microscopy and light microscopy. ResultsThe concentration of cefuroxime in aqueous humor in control group reached the highest at 0.5 h after injection of cefuroxime(47 736.18 ng/mL),while that was extremely low 24 h later(10.92 ng/mL).The concentrations of cefuroxime in aqueous humor were higher than 500 ng/mL within 7 d after implantation,and that was very low at d28(59.20?39.05 ng/mL).And the plasma concentrations of cefuroxime had been at lower levels ever since the implantation(

OBJECTIVE: To analyze the forensic pathological characteristics of sudden death caused by pulmonary thromboembolism and the chronological transformation of thrombus and explore the assessment method of the causal relationship between previous trauma and the following fatal PTE episode. METHODS: All the 23 cases reviewed here were collected from our institute files from the year of 1998 to 2008. RESULTS: Trauma, surgery and braking etc. were all risky factors of PTE. Of these cases, 12 cases were caused by trauma, 21 cases were caused by surgery and 22 cases died in hospitals which were often happened one or two weeks after injury or one week's postoperative time. Of all the cases, 6 cases had single attack of thrombus and the rest 17 cases had the recurrence of thrombus. The number of the leg deep vein to be the embolic source was 16 cases which were often seen in the left leg. CONCLUSION It is important to confirm the embolic source, trauma, surgery and chronological events in determing the sudden death with PTE.
Adolescent ; Adult ; Aged ; Autopsy ; Cause of Death ; Child ; Death, Sudden/etiology* ; Expert Testimony ; Female ; Forensic Pathology ; Humans ; Leg/blood supply* ; Male ; Middle Aged ; Pulmonary Artery/pathology* ; Pulmonary Embolism/pathology* ; Retrospective Studies ; Risk Factors ; Venous Thrombosis/pathology* ; Wounds and Injuries/complications* ; Young Adult

Objective To investigate the frequency and mutational status of JAK2V617F mutation in Chinese patients with chronic myeloproliferative disorders(CMPD) and to study the relative quantification of mutated JAK2 mRNA and the clinical significance. Methods JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation system polymerase chain reaction(ARMS-PCR), the relative quantification of mutated JAK2 mRNA was studied by using capillary electrophoresis. Results A higher prevalence of JAK2V617F in either the heterozygotc or homozyote status in essential thrombocythemia (ET) was observed in elderly patients with ET (P<0.05). The presence of JAK2V617F was found to be significantly correlated with the age at diagnosis (P<0.05); patients with age ≥ 60 years showed significantly higher JAK2 mutated RNA levels than those with age < 60 years (P<0.05); the presence of JAK2V617F in polycythemia vera (PV) and ET was found to be significantly associated with higher hemoglobin level and higher leukocyte count (P< 0.05). In addition, higher leukocyte count was observed in homozygous ET patients than in heterozygous ET patients (P<0.05). The frequency of JAK2V617F mutation and the prevalence of homozygote in PV patients were higher than those in ET patients (P<0.05). The differences of JAK2V617F mRNA levels among PV, ET and chronic idiopathic myelofibrosis (IMF) were not significant. Conclusions ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation owing to its sensitivity and along with capillary electrophoresis, quantitative assay for mutated JAK2 mRNA, diagnosis of CMPD and judgement of prognosis become possible.

This study summarized the experiences of disease-specific performance appraisal at tertiary hospitals in Shanghai,which was launched since year 2013 by Shanghai Hospital Development Center (SHDC). 38 tertiary hospitals in Shanghai were included in the study. A disease-specific performance appraisal system centering on quality and performance, by means of case-mix model, classified surgery management and typical disease screening, and leveraging disease and surgery difficulties analysis, and inter-hospital performance appraisal of typical diseases. This reform has established appraisal criteria of disease difficulty management,coding criteria and data norms,guiding such hospitals to consolidate their functional positioning of focusing on difficult,urgent and complicated cases in the medical service delivery system.All these efforts have paved the way for the reforms to build a hierarchical medical service system, pricing per disease, payment per disease, and consolidate performance appraisal of medical workers.

OBJECTIVETo observe the effect of leukemic cells on blood-brain barrier (BBB) in mice with central nervous system leukemia (CNSL) by establishing mice CNSL model and an in vitro BBB model and explore the mechanism of leukemic cell infiltrating central nervous system (CNS).

METHODSAfter splenectomy, cytoxan intraperitoneal injection, and sublethal irradiation, 10 BALB/c nu/nu mice were transplanted intravenously with 1.2 × 10(7) of SHI-1 human monocytic leukemic cells. Mice were monitored for survival and clinical manifestation of nerve palsy. The leukemic cells engrafted were examined by RT-PCR, histopathology and bone marrow (BM) smears. Immunofluorescence analysis with laser scanning fluorescence confocal microscopy was used to determine the expression of fibrinogen and tight-junction protein ZO-1. An in vitro BBB model composed of human brain microvascular endothelial cells (BMVECs) was developed on a Matrigel-based insert. Different leukemic cell lines were seeded onto the upper compartment of transwell insert. After incubated for 24 h with BMVECs, cells that had migrated into the lower compartment were counted and analyzed.

RESULTS(1) Paralysis with or without sight loss was developed in half the mice 30-35 d after innoculated with SHI-1 cells. Leukemic cells infiltrates were observed in BM and in different part of brain tissues including brain parenchyma. The transcriptions of human MLL/AF6 fusion gene were also detected in BM and brain tissues in paralysis mice. The fibrinogen expression and ZO-1 disruption were detected in the infiltrated tissue. (2) After 24 h incubation with leukemic cells, the BMVECs sheets were disrupted and grew singly and ZO-1 expression was down-regulated markedly. SHI-1 cells showed more injurious to BMVECs and higher invasive rate \[(40.33 ± 1.53)% vs (11.83 ± 1.44)%, P < 0.05\] than HL-60 cells did.

CONCLUSIONOne of the mechanisms of leukemic cells infiltrates CNS in CNSL is injure to the BBB.

Animals ; Blood-Brain Barrier ; physiology ; Central Nervous System ; pathology ; Central Nervous System Neoplasms ; pathology ; HL-60 Cells ; Humans ; Leukemia ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo investigate the influence of tissue inhibitor of metalloproteinase 2 (TIMP-2) on the infiltrative patterns of human monocytic leukemic cell line SHI-1 in nude mice.

METHODS1) 1 x 10(7) TIMP-2 gene transduced SHI-1 (SHI-1-TIMP-2) and SHI-1 transduced MSCV gene (SHI-1-MSCV) cells were inoculated via tail vein into 6-week nude mice, which pretreated by splenectomy, cytoxan intraperitoneal injection, and sublethal irradiation(referred as SCI nude mice). 30 days after inoculation, half of the mice were sacrificed, and the infiltration patterns were investigated by histological exam and human CD45 immunohistochemistry, other mice were observed for survival time. 2) Leukemic cells inoculated subcutaneously into the axillary area of mice without any pre-treatment. On day 23 and 30, mice were sacrificed to measure the volume of neoplasm. TIMP-2 protein expression and the micro vein density were detected by immunohistochemistry.

RESULTSIn SCI nude mice inoculated via caudal vein with SHI-1-TIMP-2 cells, the survival time was shorter and infiltration (including in central nervous system) was higher than that in those inoculated with SHI-1-MSCV cells. However, in inoculated subcutaneously group, the neoplasm though grew rapidly at first, over expression of TIMP-2 limited the tumor growth and angiogenesis.

CONCLUSIONThe functions of TIMP-2 are diversity; the role of TIMP-2 in tumor infiltration and metastasis was worthy of further investigation.

Animals ; Cell Line, Tumor ; DNA, Complementary ; genetics ; Genetic Vectors ; Humans ; Leukemia, Experimental ; genetics ; pathology ; Leukemic Infiltration ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Transfection

OBJECTIVETo explore the optimal methods for the reconstruction and preservation of the glans after partial penis resection in the treatment of early-stage penile cancer.

METHODSBetween January 2012 and June 2015, we treated 6 cases of early- stage penile cancer by partial penis resection, inner thigh skin graft, and glans reconstruction and followed them up for 0.5-3 years.

RESULTSThe length of the penis before and after operation was ([6.5 ± 1.2] vs [4.5 ± 1.8] cm) in the flaccid state and ([12.8 ± 2.3] vs [9.1 ± 2.1] cm) in the erectile state. The sense of the reconstructed glans was completely recovered at 3 months after surgery. The glans skin was pale red and soft, nearly normal at 12 months, with no obvious graft contracture or scar formation. All the patients achieved normal erection and their partners were satisfied with their intercourse. No recurrence or metastasis was observed.

CONCLUSIONThe strategy of partial penis resection, inner thigh skin graft and glans reconstruction, simple, effective, and with few complications, is one of the best treatments of early-stage penile cancer, which not only ensures radical removal of the tumor but also maximally reserves the function of the organ.

Humans ; Male ; Penile Neoplasms ; surgery ; Penis ; surgery ; Reconstructive Surgical Procedures ; Skin Transplantation ; Thigh