Volume 87  ·   Issue 08  ·  2007

In recent years, clinical workers at home and abroad have found that most leukemias have some kind of chromosome translocation, which can produce new fusion genes and encode fusion proteins, and these markers can be used to diagnose different types of leukemia.

China is a country with high incidence of infectious diseases, with a large number of infectious diseases, and complex and diverse types of pathogenic microorganisms. Difficult and uncultivatable microorganisms are important pathogens of infection. It will be a strong guarantee to reduce missed diagnosis of pathogenic microorganisms to pay attention to the application of cutting-edge technology in clinical microbiological examination and improve the diagnosis rate of difficult pathogens.

In recent years, the term Fast-track surgery appears frequently in the international surgical literature search data, and there are many articles on this topic, which involve the surgery in various regions, and the effect is very good, which obviously speeds up the recovery of patients and shortens the hospitalization days after surgery.

In the spring of 2003, at the beginning of the epidemic of severe acute respiratory syndrome (SARS), a large number of first-line medical staff were infected by rescuing SARS patients. Before March 24, 2003, the number of medical staff cases accounted for 27.43% of all cases.

Leukemia is a clonal disease caused by acquired hematopoietic stem cell mutation. Chromosome analysis can reveal genetic changes as malignant clonal markers, so it has been paid attention to. In recent years, with the deepening of observation and technological progress, important progress has been made in the chromosome research of leukemia. Banding karyotype analysis has become one of the conventional methods of leukemia diagnosis. A new molecular-cytogenetic technology-fluorescence in situ hybridization (FISH) is being widely used as an important supplement to the former. The new discoveries in leukemia chromosome research and the new progress in detection technology are introduced as follows.

Objective to investigate the relationship between the differential protein expression of leukemia cells before induction therapy and the prognosis of patients with acute myeloid leukemia (AML-M2a) with significant time difference in the first sustained complete remission (CCR1) by proteomic technique. Methods the extracted bone marrow mononuclear cells of 17 patients with AML-M2a before the first induction remission were grouped according to the CDR1 time after regular chemotherapy. Group A (11 cases) with more than 12 months, group B (6 cases) with less than 6 months, and group B (3 cases) with bone marrow mononuclear cells at the time of relapse were group C. The proteins of leukemia cells in groups A, B and C3 were separated by two-dimensional electrophoresis technique, and some of the differential proteins were identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS) Six differential proteins were identified by mass spectrometry, namely, specific molecular chaperone B of tubulin, myeloperoxidase, 2CH domain of transgelin, glutathione S-transferase, zinc ester protein and glyceraldehyde 3-phosphate dehydrogenase; Three differential proteins were identified by mass spectrometry in group C, namely NAD (P) H, HES1 and hypothetical protein.Conclusion the differential protein expression level of leukemic cells before induction remission therapy in AML-M2a patients is related to their prognosis, and the protein of bone marrow cells changes when leukemia relapses.

Objective to isolate the differentially expressed genes in the bone marrow of patients with familial acute myeloid leukemia, and to explore the molecular mechanism of the occurrence and development of acute leukemia at the gene level. Methods the cDNA subtraction library of familial acute myeloid leukemia was constructed by Super SMART and suppressive subtractive hybridization (SSH) techniques. Screening and identification of differentially expressed fragments by differential screening technique; The sequencing results were analyzed by BLAST homology. Results the subtracted cDNA library of familial acute myeloid leukemia was successfully constructed. Seventeen gene fragments were cloned, screened and identified, homologous to known genes, and 11 were unknown gene fragments. Conclusion SSH technique is an effective method for screening differentially expressed genes; obtaining a plurality of known genes associated with cell proliferation and differentiation; Eleven new gene EST fragments were obtained and included in GenBank.

Objective to establish a fluorescence real-time quantitative polymerase chain reaction (real-time PCR) method for the simultaneous quantitative detection of fusion genes formed by chromosome translocation in multiple leukemias, and to understand the feasibility of application of array PCR. Methods 82 primers were combined, 66 PCR parallel tubes were set up, and 125 spliced bodies formed by 37 common fusion genes in leukemia and the activation of 4 common proto-oncogenes in leukemia were quantitatively detected simultaneously. real-time quantitative PCR was performed by Eva Green fluorescent dye method, ABL gene was used as internal reference, and comparative Ct method was used for relative quantitative detection. A total of 14 fusion gene types and all 4 proto-oncogene activation were detected in 31 patients; A patient with 5 positive fusion genes and two activated proto-oncogenes was detected; Comparison with the results of multiplex nested PCR showed that the sensitivity of this protocol was slightly lower than that of nested PCR, but the difference was not statistically significant (P =0.009); The expressions of BCR/ABL fusion gene and WTl and EVIl proto-oncogene in 6 patients with CML were decreased to varying degrees after treatment, which was consistent with the clinical treatment. The results of gene quantitative analysis indicate that this scheme can quantitatively analyze the activation of fusion genes and oncogenes in common leukemia, and it has application value in the diagnosis and efficacy monitoring of leukemia. Conclusion the simultaneous quantitative detection of multiple leukemia fusion genes by PCR array is suitable for screening fusion genes in newly diagnosed patients with leukemia and detection of minimal residual disease (MRD), and it is more useful for detecting the variation of fusion genes in cases with multiple fusion genes and in the course of treatment.

Objective to compare the differential protein expression profiles of retinoic acid resistant cells and sensitive cells, and to analyze and screen the retinoic acid resistance-related proteins. Methods the whole proteins of retinoic acid resistant cell line MR2 and retinoic acid sensitive cell line NB4 were isolated by two-dimensional gel electrophoresis. The differentially expressed protein spots were screened by PDQuest v7.1 analysis software, and the peptide mass fingerprints of the differential proteins were obtained by mass spectrometry analysis. The differential proteins were identified by querying the SWISS-PROT protein database by Mascot software, and the candidate retinoic acid resistance-related proteins were obtained. The differential proteins were verified by Western blot assay and semi-quantitative RT-PCR at protein level and mRNA level. Results the two-dimensional electrophoresis patterns of APL cells with high resolution and good repeatability were obtained. The average protein spots of retinoic acid resistant cell line MR2 and retinoic acid sensitive cell line NB4 at pH 4-7 showed 890 ± 45 and 912 ± 56 respectively. 57 differential protein spots were analyzed between the two cells, and 23 protein spots were found in RA resistant cell line MR2 34 protein spots were down-regulated. 25 differential protein spots were identified by mass spectrometry, and 17 were successfully identified, corresponding to 13 different molecules. One of them was a new protein with unknown function, FLJ00279, and the rest belonged to oncoprotein (DJ-1), transcription-related protein (MYC promoter binding protein 1), molecular chaperones [heat stress protein (HSP) HSP70, HSP60 and protein disulfide bond isomerase], metabolic proteins (inhibin, triose phosphate isomerase 1 and calreticulin), signal transduction (Rho GDP dissociation inhibitor beta) and cytoskeletal proteins (ACTG1 protein, β-5-tubulin inhibitor and keratin 10). The oncoproteins DJ-1, calreticulin, HSP60 and ACTG1 were highly expressed in RA drug-resistant cell line MR2, The results of Western blot tests were consistent with the results of two-dimensional electrophoresis and showed that DJ-1 and calreticulin were differentially expressed on multiple isomers. However, semi-quantitative RT-PCR showed that the difference of protein expression between HSP70 and HSP60 was not correlated with the changes of mRNA levels. The results of the two experiments suggested that the modification or cleavage of the post-translational protein mediated the difference of protein expression. Conclusion The new protein FLJ00279 and the known functions of retinoin resistance-related proteins such as DJ-1, calreticulin, HSP70, HSP60, inhibin and MYC promoter binding protein 1 were screened by two-dimensional gel electrophoresis and mass spectrometry. Although their direct relationship with retinoin resistance needs to be further confirmed, it provides new clues for understanding the mechanism of retinoin resistance from a multi-factor perspective.

Objective to compare the value of 3D-spiral CT angiography (3D-CTA) and 3D-digital subtraction angiography (3D-DSA) in the diagnosis of intracranial aneurysm. Methods 86 patients with spontaneous subarachnoid hemorrhage (SAH) from January 2004 to March 2005 in Tianjin Huanhu Hospital were examined by 3D-CTA and 3D-DSA. The imaging characteristics of CTA and DSA were studied and compared. Results 94 aneurysms were detected in 86 patients, 12 were negative, and 94 aneurysms were detected in 74 patients. The accuracy of 3D-DSA and 3D-CTA was not statistically significant.

Objective to investigate the significance and method of 64-slice spiral CT in the screening of coronary artery disease in healthy persons. Methods 286 healthy persons over 40 years old with at least one risk factor of coronary artery disease were examined by 64-slice spiral CT. The volume data were obtained by Toshiba Aquilion 64-slice spiral CT machine. The data were obtained by TCOT algorithm. The data were examined by maximum density projection (MIP), volume reconstruction (VR), multi-plane reconstruction (MPR), CURVE reconstruction (CURVE) and three-dimensional (3D) processing, and the display of coronary artery disease was observed. Some positive patients were compared with catheter controls. Results 239 vessels were found to be narrowed in different degrees in 286 cases, of which 138 vessels were less than 50%, 82 vessels were 50% ~75%, and 19 vessels were greater than 75%. 28 cases were examined by catheter, and 12 cases were operated by stent. Conclusion 64-slice spiral CT is a rapid, effective, safe, reliable and minimally invasive examination and diagnosis method for early detection of coronary artery disease, especially for those patients with coronary artery disease who appear to be healthy and asymptomatic 64-slice spiral CT can replace coronary angiography and will be the first choice method for screening coronary heart disease.

Objective to explore the application value of trans-annular patch technique with autologous pericardium simultaneously enlarging the pulmonary annulus and valve in the reconstruction of right ventricular outflow tract (RVOT). Methods RVOT reconstruction was performed on 32 patients with congenital heart disease, including 24 cases of Fallot tetralogy, 5 cases of right ventricular double outlet method type 4, and 3 cases of severe pulmonary stenosis. Results 1 case died after operation, 6 cases were cured of other complications after operation. 31 cases were followed up for 4~40 months, and the cardiac function was grade Ⅰ ~ Ⅱ, and the pulmonary valve function was good; There were 17 cases with no or minimal pulmonary valve regurgitation, 12 cases with mild valve regurgitation, 2 cases with moderate valve regurgitation, and no need for reoperation. Conclusion the short-term and medium-term effects of reconstruction of RVOT with autologous pericardial enlargement of pulmonary valve annulus and valve technique are good, but the long-term effects remain to be observed.

Objective to evaluate the effect of surgical treatment of critical acute abdomen in low-weight newborns. Methods The clinical data of 228 neonates with critical acute abdomen weighing less than 2500 g treated by neonatal surgery in Nanjing Children's Hospital affiliated to Nanjing Medical University from January 2000 to January 2006 were retrospectively analyzed, and the relationship between gestational age, birth weight, diagnosis and treatment improvement and prognosis was compared. Results The admission age of the children was 1 h ~7 d, with an average of (4.1±0.7) d; There were 141 premature infants and 87 full-term infants; 26 cases with birth weight ≤2000 g, 202 cases with birth weight between 2000 and 2500 g; There were 83 cases from January 2000 to January 2003 (first 3 years group), 145 cases from January 2003 to January 2006 (second 3 years group), 202 cases were cured, 26 cases died (11.4%). In the premature infants group, ≤2000 g group, the mortality rate in the first 3 years group [14.89% (21/141), 53.84% (14/26), 18.07% (15/83)] was much higher than that in the full-term sample group, 2000~2500 g group and second 3 years group (χ 2=4.455, P<0.05, χ 2=52.324, P<0.01, χ 2=5.745, P<0.05). The surviving infants were followed up for 4~48 months, and their body weight and height reached the normal standards. Conclusion Premature birth and low birth weight are closely related to neonatal acute abdomen death; With the improvement of neonatal surgery, prenatal diagnosis and perioperative management, the success rate of treatment of critical acute abdomen in low birth weight neonates is increasing.

Objective to investigate the expression of type Ⅰ and type Ⅲ procollagen genes and the mRNA of SMAD (SMAD2, SMAD3, SMAD4, SMAD7) in sclerotic skin tissues of progressive systemic sclerosis (PSS), and to analyze their role in the pathogenesis. Methods the expression of type Ⅰ and type Ⅲ procollagen genes and mRNA of different types of SMAD (SMAD2, SMAD3, SMAD4, SMAD7) in sclerotic skin tissues of 11 patients with PSS and 11 normal controls were detected by fluorescence real-time quantitative polymerase chain reaction (P<0.05). The expression of SMAD4 mRNA was positively correlated with the expression of type Ⅰ and type Ⅲ procollagen genes α 1 mRNA (r values were 0.728 and 0.678, respectively, both P<0.05). The expression of SMAD3 mRNA was positively correlated with the expression of type Ⅰ procollagen gene α 1 mRNA (r =0.859, P<0.01).

Objective to investigate the clonality of peripheral papilloma of breast, its cancerous cells and intraductal carcinoma, and to find a more specific and effective auxiliary method for early diagnosis. Methods 26 cases of peripheral papilloma, 25 cases of peripheral papilloma with atypical hyperplasia, 27 cases of intraductal carcinoma and 16 cases of cancerous to intraductal carcinoma were collected as the research objects, and 20 cases of normal breast tissues were used as the control. The clonality analysis technique based on female X chromosome AR gene polymorphism was used to determine the clonality of various samples according to different bands. Results 24 cases (92.3%) of 26 cases of peripheral papilloma and 20 cases of normal breast tissues showed polyclonal hyperplasia; 27 cases of intraductal carcinoma showed monoclonal hyperplasia; Among 25 cases of peripheral papilloma with atypical hyperplasia, 10 cases (40%) showed loss of AR locus length polymorphism, which belonged to monoclonal hyperplasia; Conclusion Normal breast tissue and peripheral papilloma are polyclonal, while some cases of peripheral papilloma with atypical hyperplasia are monoclonal. Peripheral papilloma with atypical hyperplasia belongs to precancerous breast lesions. Clonality analysis can be helpful for screening high risk cases and differential diagnosis.

Objective to construct a green fluorescent protein expression vector pEGFP-C3-CagA containing Helicobacter pylori cytotoxin-related gene A (CagA) gene fragment. Methods the CagA gene fragment was amplified from Helicobacter pylori cell line, recovered, purified, ligated to plasmid pGEM-T, sequenced, digested and ligated with plasmid pEGFP-C3. pEGFP-C3-CagA was transfected into gastric cancer cell line BGC823 by liposome method, and the transfected cells were observed by fluorescence microscope. Results the sequencing and digestion proved that the insertion of CagA into the plasmid was correct, and the vector pEGFP-C3-CagA was constructed. After transfection, green fluorescence was observed in gastric cancer cell line BGC823 by fluorescence microscope. Conclusion the eukaryotic green fluorescent vector expressing CagA was successfully constructed.

Objective to evaluate the safety and efficacy of different doses of controlled release arsenic trioxide (As2O3) eluting stents in the prevention and treatment of restenosis after coronary artery stenting in dogs. Methods Polybutyl methacrylate/nano-silica-polylactic acid/glycolic acid was used as the stent coating, and naked stents, coated stents, large, medium and small doses of As2O3 (3.2, 2.4, 1.6 μ g/mm2) eluting stents were randomly placed into the circumflex or anterior descending coronary artery of dogs (6 rats in each group). The rats were sacrificed 4 weeks after operation, and the histological changes of the vascular segments of the stents were observed. Results The endothelium in each group was intact, and no obvious tissue reaction was observed. The mean neointimal thickness in the large and medium dose groups was significantly smaller than that in the small dose group, coated group and naked stent group [(0.27 ± 0.03) mm, (0.39 ± 0.06) mm, (0.33 ± 0.02) mm, P<0.01]; The area of neointima was significantly smaller than that of the small dose group, the coating group and the bare stent group [(1.93±0.29) mm2, (2.44±0.15) mm2, (2.40±0.32) mm2,P<0.01]; The stenosis rate of lumen area [(19.54±3.59) % vs. (22.18±3.3) %, P>0.05] was significantly lower than that in the low dose group, coating group and bare stent group [(36.22±5.17) %, (50.39±3.03) %, (46.88±5.85) %, P<0.01]; The lumen area (5.14±0.55) mm2 vs. (4.97±0.38) mm2,P>0.05) was significantly larger than that of the low dose group, coated group and bare stent group [(3.75±0.39) mm2, (2.62±0.22) mm2, (3.10±0.66) mm2,P<0.01]. The difference of each index between the low dose group and the coated group and bare stent group was statistically significant (P<0.01). Conclusion As2O3 controlled release eluting stent is safe and feasible, and it is dose-dependent to inhibit intimal hyperplasia after coronary stent surgery in dogs.

Objective to investigate the significance of connective tissue growth factor (CTGF) in the pathogenesis of renal interstitial fibrosis and to investigate the regulatory effect of rapamycin on it. Methods a rat model of unilateral ureteral obstruction (UUO) was established. The expression of CTGF mRNA and protein in the obstructed kidney and sham-operated group was dynamically observed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The expression of CTGF mRNA and protein in the model group and the rapamycin group was significantly increased (P<0.01). Compared with the model group, the expression of CTGF mRNA and protein in the rapamycin group was decreased (P<0.05), the expression of CTGF protein was down-regulated (P<0.05) and (P<0.01), and the expression of fibronectin was also down-regulated (P<0.05). Conclusion the expression of CTGF mRNA and protein in the kidney was significantly increased after ureteral obstruction.

Repair and regeneration are common phenomena in the biological world. When a tissue or organ of an organism is damaged or lost, a new tissue or organ can be formed under certain conditions. The two tissues are almost identical in structure and function. This process is called regeneration.

With the improvement of living standard, obesity has become a social problem. In the past period, the medical community often recommended low-protein, low-fat and high-carbohydrate diets to control body weight. At present, low-carbohydrate diets are still common, and there are some supportive opinions. According to the recent research results, a recommendation on diet and lifestyle has been formulated in the United States, but the long-term safety of such diets is still unclear, that is, whether low-carbohydrate diets may be related to coronary heart disease.

The incidence of ventilator-associated pneumonia (VAP) in critically ill patients undergoing mechanical ventilation is about 20%. Compared with patients without VAP, patients with VAP have a higher mortality rate, stay in ICU for a longer time, and consume more medical resources.

Parkinson's disease is one of the common diseases that cause disability in the elderly, which can lead to a significant decrease in the quality of life. Aiming at the chemopathology of Parkinson's disease and the synthesis and metabolism of dopamine, therapeutic drugs have been developed for all aspects: dopa replacement therapy (levodopa), dopamine agonist (amantadine), catechol-O-methyltransferase inhibitor, etc. However, the efficacy of these drugs can only last for several years at most.

The 14th National Pediatrics Academic Exchange Conference sponsored by the Chinese Medical Association was held in Nanning, Guangxi Zhuang Autonomous Region from November 26 to 30, 2006. The theme of this conference was "Concerning Children's Health and Creating a Harmonious Environment".

A 27-year-old male was admitted to the hospital due to dysuria for 1 month after lithotripsy of right ureteral stones. The patient underwent "holmium laser lithotripsy of right upper ureteral stones under ureteroscopy" in our hospital 1 month ago. During the operation, the urethral mucosa was obviously damaged and strictured, and the old patchy injury around the right ureteral orifice was seen (retrograde intubation angiography before operation), which was considered to be complicated with urethral stricture.