Volume 86  ·   Issue 32  ·  2006

human leucocyte antigen (HLA) system has been discovered, understood and applied for nearly half a century, and the continuous and in-depth reveal of this system has greatly promoted and improved the backward situation of basic immunology, organ transplantation, bone marrow bank/cord blood bank, human genetics, forensic medicine, tumor immunology, tumor vaccinology, genetic susceptibility to autoimmune diseases and infectious diseases [1].

myeloproliferative disorders (MPD) are diseases with abnormal proliferation of hematopoietic cells or connective tissue cells. Although this disease has been discovered for nearly 100 years, the etiology is still unclear. There is no clinical diagnostic method that can be called gold standard, and there is no recognized optimal treatment.

In recent decades, although chemotherapy has made great progress in the treatment of leukemia, about 70% of acute myeloid leukemia (AML) and 80% of acute lymphocytic leukemia (ALL) can achieve complete remission (CR), there are still 20% ~30% of initially treated cases who cannot achieve CR (refractory leukemia), and a considerable number of patients who achieve CR eventually relapse.

1. Unhealthy living habits and behaviors are seriously harmful to life and health. The Shanghai hepatitis A pandemic, which occurred in 1988 and infected more than 300,000 people, was caused by eating raw or half-raw cockles contaminated with hepatitis A virus; Infectious atypical pneumonia (SARS), which occurred in southern China in winter and spring from 2002 to 2003, spread rapidly to 26 provinces, autonomous regions, municipalities directly under the central government and 29 countries and regions in China, and shocked the world, is closely related to slaughter and culinary civet cat; In recent years, most cases of human pneumonic plague occurred in China were caused by capturing and stripping the fur of marmots without protection measures; The only cause of human infection with type Ⅱ Streptococcus suis occurred in Sichuan and other provinces of China in 2005 was slaughter, treatment of diseased and dead pigs. 　　

Objective to investigate the inhibitory effect of simvastatin on endothelial cytotoxicity and inflammation induced by homocysteine (HCY). Methods Human umbilical vein endothelial cells were treated with different concentrations of HCY (0.1, 0.25, 0.5, 1 mmol/L) for 24h, and the cell viability was detected by tetrazolium blue. Human umbilical vein endothelial cells were pretreated with simvastatin (1, 10, 20 μ mol/L) for 1 h, and then co-incubated with HCY (0.25 mmol/L). The cell viability was detected by MTT, and the expression of related inflammatory factor proteins at different time points was analyzed by Western blot and ELISA. Results After HCY treatment, the survival rate of human umbilical vein endothelial cells was significantly reduced; After pretreatment with simvastatin (1, 10, 20 μ mol/L), the activity of endothelial cells were 47%±4%, 68%±6%, 89%±6%, respectively, which were significantly higher than those of HCY (0.25 mmol/L) alone (22%±3%, all P<0.01). The expressions of TNF-α, IL-6, MCP-1 and ICAM-1 in endothelial cells induced by HCY were 0.23±0.05, 0.14±0.03, 0.13±0.04, 0.21±0.07, respectively, and the differences were not statistically significant compared with those at 0 h. Conclusion Simvastatin has a significant inhibitory effect on homocysteine-mediated endothelial cell injury and inflammatory response.

Objective to investigate the effects of autologous blood washing and retransfusion on erythrocyte immunity and renal function during open heart surgery with cardiopulmonary bypass. Methods Thirty-two patients with heart valve replacement were divided into the experimental group with autologous blood washing and retransfusion and the control group with remaining blood and reserved erythrocyte retransfusion (16 cases each). During cardiopulmonary bypass, blood loss was recovered, washed and retransfused with a blood processor. Peripheral venous blood anticoagulated with heparin was collected before operation, 12 h, 24 h, 72 h and 7 d after operation, respectively. The erythrocyte immune functions, namely, erythrocyte C3b receptor rosette rate (C3bRR), erythrocyte immune complex rosette rate (RBCICR), plasma free hemoglobin content (FHB), urine protein and creatinine clearance rate (Ccr), were compared between the two groups. Results (1) RBG-C3bRR (14.3%±4.7%, 15.9%±3.6%, 16.6%±2.8%, 19.9%±4.1%) and RBC-ICR (8.7%±1.9%, 9.2%±2.0%, 9.5%±2.6%, 12.0%±2.0%) in the experimental group were significantly higher than those in the control group at 12, 24, 72 hours and 7 days after operation (all P<0.05). (2) FHB (0.41g/L±0.13g/L, 0.03g/L±0.02g/L) in the experimental group was significantly lower than that in the control group (1.02g/L±0.23g/L, 0.54g/L±0.09g/L) at 12 hours and 24 hours after operation (P<0.05)

Objective to investigate the protective effect of intensity-modulated radiotherapy on parotid function in patients with nasopharyngeal carcinoma and the relationship between the change of parotid function and dose-volume. Methods From August 2002 to December 2004, 48 patients with nasopharyngeal carcinoma who received intensity-modulated radiotherapy were treated with 99mTc clearance rate before, at the end of and 3 months after treatment, respectively, to determine their secretion index (EI) and uptake index (UI), and analyzed with the dose-volume histogram (DVH) of parotid gland. Results The mean dose of parotid gland on the healthy side and the affected side were (22.8±4.5) Gy and (31.9±4.1) Gy, respectively. The symptoms of dry mouth were mild in the whole group. The EI value of parotid gland on the healthy side was 0.35±0.25 before radiotherapy, 0.31±0.24 at the end of radiotherapy, and 0.33±0.22 at 3 months after radiotherapy. The UI value of parotid gland on the affected side was 7.12±3.56, 5.81±2.25 and 5.72±2.81 at 3 time points, respectively, with no statistical significance. The EI and UI of parotid gland on the affected side were 0.36±0.27 and 8.02±3.89 before radiotherapy, and 0.21±0.16 and 4.87±2.45 at the end of radiotherapy, respectively. There was a significant difference in the decrease of EI between V25 (parotid gland volume with exposure dose<25 Gy) ≥50% and V25<50% (P<0.01), but no significant difference in the decrease of UI between the two cases (P>0.05). Conclusion The parotid gland dose below 26 Gy has a threshold effect on the functional protection of parotid gland, and there are dose and volume thresholds in the functional protection of parotid gland.

Objective to observe whether transfection of hypoxia-inducible factor 1 α (HIF-1 α) can enhance the invasion ability of human prostate cancer cells in vitro, and to explore its molecular mechanism.Methods the recombinant eukaryotic expression vector pCDNA3.1 (-) /HIF-1 α was packaged with Lipofectamine2000 and transfected into human prostate cancer cells LNCaP, and the resistant clones stably expressing HIF-1 α were screened at 600 μ g/ml G418. The overexpression of HIF-1 α was identified by immunofluorescence and Western blot, and the expression of invasion-related proteins E-cadherin, vimentin, matrix metalloproteinase-2 (MMP-2), cathepsin D (cathepsin D) and urokinase-type plasminogen activator receptor (uPAR) was detected by Western blot. The invasion ability of the cells was verified by Transwell. Results Compared with untransfected LNCaP, the HIF-1 α protein band appeared in transfected cells LNCaP/HIF-1 α, and the expression of E-cadherin was deleted, while the expression of vimentin, MMP

Objective to establish a rapid and accurate molecular biological method for the detection of Yersinia enterocolitica in diarrhoeal feces, to replace the cumbersome and time-consuming method of culture and identification. Methods the specific primers and probes designed for the yst gene on the chromosome of pathogenic Yersinia enterocolitica were used to detect pathogenic (7 serotypes) and non-pathogenic (5 species) Yersinia enterocolitica, other species of Yersinia (8 species) and other intestinal bacteria (8 species) by real-time quantitative PCR. In addition, 200 cases of diarrhoeal feces identified by culture were detected by real-time quantitative PCR, and the agreement between the two methods was compared. Results the primers and probes were 100% specific for pathogenic Yersinia enterocolitica, and there was no cross-reaction with other bacteria. The lowest detection concentrations in pure culture and feces were 102 colony forming units (CFU) /ml and 103 CFU/g, respectively. Among 200 samples, the same 18 positive samples were detected by culture identification method and real-time quantitative PCR, and the agreement degree was 100%. Conclusion Real-time quantitative PCR reaction is highly specific for pathogenic Yersinia enterocolitica and has good agreement with culture identification method, is convenient and quick, and can be applied to clinical detection.

Objective to investigate the regulatory effect of D-α-tocopherol on the expression of hexokinase (HK) and its isoenzymes in human peritoneal mesothelium cells (HPMC) induced by high glucose. Methods HPMC were isolated and cultured by trypsin digestion, and the cultured cells were identified by immunocytochemical staining and scanning electron microscopy.The third generation HPMC was divided into normal control group (0.1% glucose, equivalent to 5.5 mmol/L), high glucose group (0.5%, 1.0%, 1.5%, 2.5%, 4.25% glucose) and D-α-tocopherol intervention group.After 24 h culture, the activities of HK and its isoenzymes were detected by glucose 6-phosphate dehydrogenase coupled colorimetry and temperature sensitivity test; Intracellular localization of HK protein was detected by immunocytochemical staining; The expression of HK protein and mRNA were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR), respectively. The glucose consumption in culture medium was detected by hexokinase method as the index of net glucose utilization of cells. Results Keratin and vimentin were positively expressed in the cytoplasm of HPMC, and there were abundant microvilli on the cell surface of HPMC. The total activity of HK was up-regulated in the 1. 5%, 2.5% and 4.25% glucose group, and the relative activity of HK was 115.4%, 129.1% and 155.2%, respectively, compared with the normal control group (20.6±2.5) U/g protein, and the main increase was HK Ⅱ (P<0.05). The total activity of HK was down-regulated in the 50 mmol/ld-α-tocopherol intervention group, which was 82.1% (P =0.001), and HK Ⅱ was mainly inhibited; After D-α-tocopherol intervention, the net utilization of glucose by HPMC decreased from (25.3±3.9) mmol/L to (17.3±2.1) mmol/L (P =0.018). With the increase of glucose concentration (glucose ≥1.5%), HKⅡ accumulated at the perinuclear area and was thoroughly stained, appearing as bead chain; Conclusion D-α-tocopherol inhibits the upregulation of HKⅡ activity, protein and mRNA expression by high glucose, and reduces the net utilization of glucose by HPMC. It may become one of the means to increase peritoneal dialysis ultrafiltration.

Objective to observe the curative effect of different reconstruction methods on ureteral mucosal exfoliation and explore a new method for the treatment of ureteral mucosal exfoliation. Methods Twenty adult breed dogs were randomly divided into four groups to make the ureteral mucosal exfoliation model. The mucosal defect was treated with bladder mucosa or peritoneal implantation and ureteral mucosal replacement in situ, respectively, while the control group was only placed with stent tube. Intravenous pyelography (IVP) and pathological examination were performed 10 weeks after operation. Results the control group had ureteral lumen atresia or severe stenosis. There was no obvious stenosis and dilation of ureteral lumen in the ureteral mucosal replacement in situ group and peritoneal replacement group, and the new mucosa in the reconstructed segment grew well, close to the normal ureteral structure. In the bladder mucosal group, one died, two reconstructed segments had mild upper ureteral dilation, and two ureteral lumens had no obvious stenosis. Conclusion Ureteral mucosal replacement in situ or peritoneal reconstruction with stent tube adequate drainage is ideal for the treatment of ureteral mucosal exfoliation. Bladder mucosa can be used as an alternative repair material for reconstruction of ureteral mucosa after exfoliation.

Objective to study the effects of leptin on gastric motility during interdigestive phase (MMC) and gastric motility during digestive phase in rats, and to observe the changes of serum leptin concentration in different time courses between fasting MMC and postprandial gastric motility and the relationship with motilin changes. Methods Fifty Wister rats were randomly divided into five groups for chronic experiment. Stress sensors were implanted in gastric body, gastric antrum and pyloric serosa to record gastric motility in conscious rats. Chronic venous cannula was built into external jugular vein for injection of drugs and blood samples. Serum leptin and motilin were determined by radioimmunoassay, and standard meal was used for eating meals. Results Typical interdigestive MMC movements occurred in fasting gastric motility, which were divided into Ⅰ, Ⅱ, Ⅲ and Ⅳ phases, and phase Ⅲ could be transmitted to pylorus. After eating, gastric motility showed continuous and regular peristaltic contraction during digestive phase. The serum leptin concentrations in fasting MMC were 5.6 ± 1.0 μ g/L in phase Ⅰ, 11.4 ± 2.1 μ g/L in phase Ⅱ and 2.1 ± 0.7 μ g/L in phase Ⅲ, which showed that the leptin concentration in phase Ⅱ of MMC was the highest (P<0.01 In contrast to leptin, the concentration of motilin was the highest in MMCⅢ phase, but the concentration was lower after prandial. Intravenous perfusion of leptin (2.5 μ g · kg-1.10 min) can change fasting gastric motility to a type of gastric relaxation, gastric antrum and pyloric contraction after prandial. Anti-leptin serum, atropine and cholecystokinin (CCK) -A receptor blocker valeramic acid can block the effect of leptin on gastric motility. Conclusion Leptin is a hormone that initiates fasting gastric MMCⅡ phase activity and postprandial digestive motility, and its changes can lead to delay of gastric emptying related to satiety. The effect of leptin on gastric motility is mediated by cholinergic nerve and CCK-A receptor.

Objective to study the typing of human leukocyte antigen (HLA) class Ⅰ and Ⅱ by using gene chip technique. Methods According to the common HLA-Ⅰ and Ⅱ gene loci in northern Chinese Han population and the practical needs of typing resolution in clinical application, a specific oligonucleotide medium resolution typing probe was designed and made into a genotyping chip. The polymorphic regions of HLA-Ⅰ and Ⅱ antigens were amplified by fluorescence-labeled primers with suitable PCR method, and the products were hybridized with the probes on the chip. The hybridization results were determined by fluorescence scanning and the positive probes were analyzed by specific software, so as to determine the genotypes of the samples. A total of 30 clinical samples were detected. Results All samples were successfully genotyped for HLA-Ⅰ and Ⅱ antigens. The resolution probes could distinguish 598 and 511 class Ⅱ antigen alleles, 57 and 30 specific for Ⅰ and Ⅱ antigens. Conclusion Gene chip is feasible for HLA-Ⅰ and Ⅱ antigen typing.

Objective to investigate the possible gene rearrangement of mixed lineage leukemia (MLL) in patients with acute myeloid leukemia (AML) and to explore its clinical significance. Methods Fluorescence in situ hybridization (FISH) was used to analyze 58 patients with AML (47 adult AML and 11 children AML) using multiple site-specific DNA probes (full chromosome staining, special locus, two-color or multi-color translocation fusion probes) on the basis of routine cytogenetic analysis. Results MLL gene rearrangement occurred in 6 cases, and MLL gene translocation, replication and loss occurred respectively. The positive rate accounted for 10.3%. Among them, 4 cases were adult AML and 2 cases were children AML. Besides MLL gene rearrangement, 5 patients with AML had complicated karyotype changes, namely: 47-49, XX, der (1) t (1; 17) (p6.1; q23), +4, +10, der (11) t (11; 17) (q23; q23), -17, -18, +20, +21?. ish +21 (wl 46, XX, del (5) (q13q33), r (11) (p15q25), + r (11) (p15q25). ishr (11) (wcp11+, MLL +), + r (11) (wcp11+, MLL +); 46, XY, del (11) (q23) [2] /46, idem, add (16) (p13.1) [8] /46, XY [10]. ishadd (16) (wcp16+), rea (11) (wcp11+); 55, XY, + markers.ish 11q23 (ML1x3), +21 (wcp21+); 46, XY, add (11) (q23) [6] /46, idem, t (15; 17) (q22; q21) [12] /46, XY [2]. ish dup (11) (MLL + +), t (15; 17) (PML +, RARa +; RARa-) [24]. Conclusion MLL gene rearrangement is often associated with acute myeloid leukemia, and the positive rate in this experimental group is over 10%. Because patients with MLL gene rearrangement are insensitive to conventional chemotherapy and have poor prognosis, MLL gene rearrangement should be examined routinely in patients with newly treated acute leukemia.

Objective to investigate the effect of Imatinib mesylate (STI571) on the development of dendritic cells (DC) in chronic myeloid leukemia (CML) and to explore its possible mechanism.Methods CML bone marrow mononuclear cells were isolated and cultured with recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) and recombinant human interleukin-4 (rhIL-4) in the culture system (control group) and STI571 in the experimental group. On the 8th day, recombinant human tumor necrosis factor d (rhTNF-α) was added to further stimulate the maturation. The cell morphology, phenotype and mixed lymphocyte response were observed after culture, and the concentration of vascular endothelial growth factor (VEGF) and the activity of nuclear factor κ B (NF-κ B) in the culture supernatant were detected. Results Both groups showed typical dendritic cell morphology. The expression of CD80, CD86, CD83 and human leukocyte antigen DR (HLA-DR) in the experimental group and the ability to stimulate lymphocyte proliferation were significantly higher than those in the experimental group The concentration of VEGF in the experimental group was significantly lower than that in the control group (P<0.05), while the activity of NF-κ B was significantly higher than that in the control group (P<0.05). Conclusion STI571 can promote the activation of CML bone marrow-derived DC, which may be related to the relief of the inhibition of CML-derived VEGF on DCNF-κ B activity.

Objective to investigate the mechanism of berberine, a novel p210 bcr/abl inhibitor, inducing apoptosis in human chronic myeloid leukemia cells. Methods Ph + human chronic myeloid leukemia cell line K562 expressing endogenous p210 bcr/abl protein was cultured, and the cells were interfered with berberine at the specified time and dose. The percentage of apoptotic cells was quantitatively analyzed by Annexin-V-Fluos/propidiumidide (PI) kit and flow cytometry; cytoperm/cytofix and aspartate-specific cysteine protease-3-McAb-PE were used to quantify the percentage of cells containing activated aspartate-specific Caspase-3; The phosphorylation of p210bcr/abl protein was quantitatively analyzed by immunoprecipitation (c-abl antibody) and Western blot (p-Tyr (pY99) antibody); The total amount of p210 bcr/abl protein was directly detected by Western blot (c-abl antibody); The results showed that low dose of berberamine could significantly inhibit the phosphorylation of p210bcr/abl in leukemia cells after treatment with berberamine at 8 μ g/ml for 48 h, the phosphorylation of p210bcr/abl protein in leukemia cells was only 8.41% of that in the control group, but the total amount of p210bcr/abl protein was not changed. Berberamine could also directly down-regulate the level of p210bcr/abl chaperone Hsp90 protein: the level of Hsp90 in leukemia cells treated with berberamine at 8 μ g/ml for 24 h was only 18.37% of that in the control group, and it had no significant effect on the level of Hsp70 protein which could induce apoptosis resistance in leukemia cells. Conclusion (2) Different from geldemycin (GA), the known Hsp90 inhibitor, berberamine can directly down-regulate the level of Hsp90 protein, but has little effect on the expression of Hsp70 protein, which is related to the apoptosis resistance of tumor cells. This suggests that berberamine may be a novel protein chaperone Hsp90 inhibitor, which is worthy of further study.

A 47-year-old female was admitted to Zhejiang Quhua Hospital due to abdominal mass found during routine physical examination 2 days ago. The patient had no abdominal pain, diarrhea, nausea and vomiting since her illness. No fear of cold and fever, no cough and shortness of breath, no chest tightness and palpitations.

The release of cytochrome C from mitochondria is a key step in apoptosis. In the presence of adenosine deoxytriphosphate (dATP), cytochrome C released into the cytoplasm can bind to apoptosis-related factor 1 (Apaf-1) to form multimers, and promote the binding of caspase 9 to form apoptotic bodies. caspase-9 is activated, and activated caspase-9 can activate other caspases such as caspase-3, thus inducing apoptosis.

Drinking alcohol has always been considered as a bad lifestyle that harms health, and adults are often advised to abstain from drinking in hospitals to reduce the risk of coronary heart disease. However, in recent years, some foreign studies have shown that drinking alcohol may reduce the incidence of coronary heart disease, but this statement has been controversial. Recently,<The British Medical Journal>June published the results of a large retrospective study on the relationship between drinking patterns and coronary heart disease (BMJ, 2006, 332:1244-1248).

A 10-month-old male was seen for fever and cough for 2 days. The child was the fourth child of his parents, delivered naturally at full term, with a history of asphyxia at birth, unknown APPAR score, birth weight of about 4 kg, length of 45cm, unable to raise his head or sit alone after birth to the doctor, unable to speak, and poor intellectual development. He was easy to cry, hoarse and weak in crying, and had difficulty in feeding. Vaccination was carried out on time. Physical examination: poor development, moderate nutrition, clear mind, mental tiredness, pale complexion, special face, poor response to the outside world, and pale skin all over the body.

The advent of new molecular targeting drugs has brought new opportunities to patients with advanced breast cancer, which have been widely used in clinical practice and have achieved certain curative effects.

acute monocyte leukemia (M5) has its unique karyotype characteristics and clinical manifestations [1]. Hematology researchers have been trying to find new methods to treat M5.