Volume 86  ·   Issue 26  ·  2006

In recent years, with the influence of human factors on living environment and the expansion of longevity population, the overall incidence of malignant tumors is increasing year by year, and the cases of cancer of unknown primary (CUP or cancer of unknown origin, CUO) are also common. However, there are few reports on epidemiology, diagnosis and treatment related to CUP, which is a difficult point in clinical diagnosis and treatment. CUP mostly uses empirical treatment, because of the blindness of treatment, poor curative effect, large side effects and unsatisfactory prognosis. Therefore, how to make a clear diagnosis and give active, scientific and reasonable treatment in a short time without delaying the disease condition and saving medical resources has important clinical and social significance.

Objective to observe the incidence, classification and associated malformations of aortic coarctation (CoA), and to explore its diagnosis and treatment. Methods 96 cases of CoA admitted to Pediatric Hospital of Fudan University in the past 15 years were retrospectively analyzed, including the incidence, pathological classification, associated malformations, diagnosis, treatment and prognosis. Results the proportion of male to female in 96 cases was 1.7: 1, 60% of infants under 6 months, and the percentage of congenital heart disease in the same period increased year by year, from 0.5% (5/924) in 1990 to 4.3% (15/350) in 2005. Among them, 93 cases were coarctation at the distal end of the left subclavian artery directly facing the duct arterial or near the duct arterial, 2 cases were coarctation located between the left common carotid artery and the left subclavian artery, and 1 case was located at the opening of the left subclavian artery. CoA alone accounted for only 14%, 49% were associated with ventricular septal defect (ventricular defect) or patent duct arterial, and 23% were associated with ventricular defect and patent duct arterial. Other associated malformations included aortic valve malformation or stenosis, complete atrioventricular septal defect, etc. 89 cases were diagnosed by echocardiography, and 7 cases were missed Two cases died (mortality rate 3.8%), of which 7 cases were staged surgery. They were older infants with ventricular defect and severe pulmonary hypertension or small infants with poor general condition, and there were no deaths. Recent postoperative echocardiography was satisfactory. Conclusion The incidence of CoA in China is not obviously different from that in western countries. Coarctation mainly occurs near the distal ductus arteriosus of left subclavian artery, often accompanied by ventricular septal defect and/or patent ductus arteriosus; Echocardiography is the first choice of examination, but cardiac catheterization is required if necessary; One-stage radical surgery can be performed for infants and young children, but staged surgery can be performed for children with ventricular deficiency and pulmonary hypertension and poor general condition.

Objective to investigate the method and effect of cultured pancreatic endocrine precursor cells. Methods the neonatal rat pancreatic tissue was cultured in medium containing high sugar concentration (33.3 mmol/L) and serum to obtain proliferative cells; Then transferred to lower sugar concentration (13.2 mmol/L) serum-free medium for 7 d; The cultured cells were identified by electron microscopy, dithizone staining, immunocytochemistry, radioimmunoassay (insulin assay) and the effects of cultured cells on blood glucose in vivo were observed by nude mouse diabetic model. Results the cultured cells were positive for pancreatic endocrine precursor cells labeled with neuroghrelin 3; The ratio of positive cells stained with dithizone increased from 10%±3% to 32%±6%, and the insulin secretion capacity increased from (22.7±3.6) μ U/ml to (26.0±2.2) μ U/ml, P<0.05, and the glucose concentration was 20 mmol/L. In vivo experiments showed that the cultured cells could prolong the survival time of nude mice diabetic model (10.0d vs 5.6d, P<0.05). Conclusion The culture medium with specific glucose concentration is beneficial to the culture of pancreatic endocrine precursor cells, and the low glucose medium is beneficial to the differentiation and maturation of endocrine precursor cells.

Objective to evaluate the sensitivity and specificity of myocardial radionuclide imaging (MPI) with redistributed imaging agent 99 Tcm azoethide (N-NOET) in the diagnosis of coronary artery disease. Methods Coronary angiography and 99TcmN-NOET adenosine stress test (MPI) were performed simultaneously in hospitalized patients. The total amount of adenosine was 840 μ g/kg, and the intravenous pumping was performed at uniform rate for 6 min. After 3 min of adenosine pumping, the dose of 99TcmN-NOET was 740MBq (20 mCi) intravenously. Stress myocardial imaging was performed 15 min later, and redistributed myocardial imaging was performed 2 h later. Results Fifty-three patients, aged 35-79 (56±10) years old, 36 males and 17 females were enrolled. There were 22 cases in the group without obvious coronary artery stenosis, 6 cases were positive for MPI, 16 cases were negative for MPI. The sensitivity of MPI in 99TcmN-NOET adenosine stress test for coronary artery stenosis over 50% was 100%, the specificity was 73%, the positive predictive value was 84%, and the negative predictive value was 100%. 9 patients with coronary artery disease underwent revascularization treatment, 6 patients had restenosis in the branch vessel or coronary artery stent, and 5 patients had poor perfusion in the corresponding area; Conclusion the sensitivity and specificity of 99TcmN-NOET MPI in the diagnosis of coronary artery stenosis are high, and the examination can be performed within the same day, which has good clinical application value.

Objective to analyze the correlation between admission time (weekdays OR weekends) and hospital mortality of patients in intensive care unit (ICU). Methods The clinical data of 1840 patients admitted to ICU of Peking Union Medical College Hospital from July 26, 1999 to December 30, 2002 were retrospectively reviewed, and the risk factors of hospital mortality were analyzed by logistic regression method. Results The mortality rate of patients admitted on Friday (OR 1.801, 95% CI 1.101~2.945, P =0.019), Saturday (OR 2.569, 95% CI 1.485~4.445, P =0.001) and Sunday (OR 2.542, 95% CI 1.377~4.691, P =0.003) was significantly higher than that of patients admitted on Thursday (OR 2.542, 95% CI 1.377~4.691, P =0.003). The difference in hospital mortality rate was statistically significant after adjustment by APACHE Ⅱ diagnostic classification weight, but after multivariate adjustment, the mortality rate of patients admitted on Friday (OR 1.191, 95% CI 0.628~2.259, P =0.593), Saturday (OR 1.107, 95% CI 0.528~2.324, P =0.787) and Sunday (OR 0.571, 95% CI 0.240~1.360, P =0.206)

Objective to evaluate the prognostic value of the number of naive cells in different proportions of bone marrow at 48 hours after the end of induction therapy and whether complete remission was achieved at the end of the first course of induction therapy in the treatment of acute myeloid leukemia (AML) in children. Methods 61 newly diagnosed children who entered the AML-XH-99 regimen were examined by bone marrow morphology at 48 hours after the end of the first course of induction therapy and when complete remission was achieved at the end of the first course of induction therapy. The survival rate of each group was compared by Log-rank test; The biological characteristics were compared by χ 2 test or Fisher's exact probability method (two-tailed); The COX hazard ratio model was used to evaluate independent prognostic factors. Results Bone marrow naïve cells ≥0.15 (n =11) at 48 h after the end of induction therapy were compared with<The difference in 5-year event-free survival (EFS) rate of children with 0.15 (n =35) was statistically significant (18%±15% and 49%±11%, P =0.0079); The proportion of naïve cells in bone marrow at 48 hours after the end of induction therapy was significantly correlated with whether the first course of induction therapy reached complete remission (P =0.000 028 8). The presence of morphologically recognizable naïve cells in bone marrow at the end of the first course of induction remission therapy reached complete remission (0.00<naive cells<The 5-year EFS rate of children (n =36) was 53% ± 10%, while the children (n =3) without morphologically recognizable naive cells (naive cells =0.00) were all long-term survival for 5 years. There was a statistically significant difference in 5-year EFS rate between children with complete remission at the end of the first course of induction remission therapy (n =39) and those with incomplete remission at the end of the first course of therapy (n =22) (54% ± 10% and 10% ± 9%, P =0.0002); Multivariate analysis showed that complete remission at the end of the first course of induction therapy and central nervous system leukemia at the onset of the disease had independent prognostic value (risk ratio 4.007, 7.050, 95% confidence interval 1.019-6.613, 0.018-0.547, P value 0.045, 0.008, respectively). Conclusion Whether complete remission at the first course of induction therapy has independent prognostic value; the number of naive bone marrow cells at 48 hours after the end of induction therapy can be used to predict whether children can achieve complete remission at the first course of induction therapy, so as to take timely intervention measures to improve the complete remission rate and long-term EFS rate in children with AML.

Objective to investigate the anti-Staphylococcus aureus effect of transgenic mice. Methods the recombinant plasmid of β-defensin-2 (HBD-2) was constructed by molecular cloning method. The recombinant plasmid was injected into female prokaryotic cells of mice by microinjection technique. The integration of HBD2 gene in mice was detected by polymerase chain reaction (PCR), and the expression and tissue distribution of HBD2 in transgenic mice were detected by immunohistochemistry. The whole body condition and death number of transgenic positive mice were observed by peritoneal challenge with Staphylococcus aureus. Results DNA sequence determination showed that the recombinant plasmid was correctly inserted into HBD-2 cDNA fragment. PCR showed that 7 mice had integrated HBD-2 gene, and immunohistochemistry of PCR-positive mice showed that HBD2 gene was widely expressed in lung, intestine, liver, kidney, testis, cerebellum and other tissues.

Objective to investigate the transdifferentiation of pulmonary artery endothelial cells (PAEC) into smooth muscle-like cells under hypoxic conditions and the role of myocardin in the transdifferentiation process.Methods Primary pulmonary artery endothelial cells were purified and isolated by immunomagnetic bead method (using platelet endothelial cell adhesion molecule 1 as immunosorting marker). psimyocardin RNA expression vector was constructed by RNA interference (RNAi) technique to inhibit the expression of myocardin gene.The cells were divided into normoxic group (containing 21% O2, 5% CO2 and 74% N2), hypoxic group (containing 1% O2, 5% CO2 and 94% N2), normoxic + myocardin inhibitory group and hypoxic + myocardin inhibitory group, and cultured for 1 d, 4 d and 7 d, respectively.The expression of myocardin mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR); The expression of α-smooth muscle (α-SM) actin was detected by immunofluorescence, and the transdifferentiation of smooth muscle-like cells was identified by morphology. Results (1) The positive rate of α-SM-actin in PAEC was significantly lower in the 4 d hypoxia group (0.96‰±0.08‰) than that in the 7 d group (2.07‰±0.06‰, P<0.01), and the positive cells were spindle or polygonal, but there was no positive expression of α-SM-actin in the normoxia group and the 1 d hypoxia group; (2) RT-PCR showed that the expression of myocardin mRNA was significantly lower in the hypoxia 4d group (0.14±0.01) than in the hypoxia 7d group (0.23±0.03, P<0.01), but the expression of myocardin mRNA was not detected in the normoxia group and the hypoxia 1d group. (3) After RNAi silenced the expression of myocardin gene, the mRNA level of myocardin mRNA was significantly lower than that in the hypoxia 4d group (0.03±0.02) and the hypoxia 7d group (0.05±0.01, P<0.01); The transdifferentiation rate of smooth muscle-like cells was also significantly reduced (0.19‰±0.07‰,0.21‰±0.04‰,P<0.01). The above was repeated three times in each group.Conclusion There are a few cells in PAEC that have the potential to transdifferentiate into smooth muscle-like cells, and hypoxia can significantly promote their transdifferentiation, and myocardin plays an important role in this transdifferentiation process.

Objective to study a group of patients with primary liver cancer (HCC) treated with transhepatic arterial chemoembolization (TACE) sequential combined with CT-guided radiofrequency ablation (RFA) and TACE sequential combined with CT-guided RFA and cytokine-induced killer cell (CIK), and to evaluate the efficacy and antiviral effect of the two methods in the treatment of primary liver cancer. Methods 195 patients with primary liver cancer were diagnosed in Guangzhou Sun Yat-sen University Cancer Hospital from July 2002 to December 2005. After comprehensive judgment of enhanced CT scan, PET-CT, digital subtraction angiography (DSA) and/or pathological biopsy combined with clinical examination results, 64 patients met the enrollment criteria. 64 patients with primary liver cancer were divided into groups 6 to 8 weeks after TACE sequential combined with RFA treatment. After clinical evaluation, there was no clear residual lesion, and 33 patients in the combined CIK treatment group were first completed 4 consecutive CIK cells transfusion through vein/hepatic artery, with an interval of 1 to 3 weeks each time; The patients were followed up for more than 1 year and the tumor status was evaluated every 1~2 months. The tumor status of 31 patients in the TACE combined with RFA group was evaluated every 1~2 months after discharge and followed up for more than 1 year. Results the 1-year survival time of both groups was 100%. There were 2 cases of recurrence between June and 1 year, 1 case had intrahepatic recurrence, 1 case had hilar lymph node metastasis, and 29/33 cases had tumor-free survival (90.19%). In the TACE combined with RFA treatment group, 2 cases had recurrence within 6 months, and 6 cases had recurrence between June and 1 year, of which 5 cases had intrahepatic recurrence, 1 case had lung metastasis, and 23/31 cases had tumor-free survival (68.01%). After 6 months follow-up, there was no statistically significant difference in 1-year tumor-free survival between the two groups (P =0.1309). In the combined CIK group, the HBV DNA content was less than 1.0×103 before treatment in 19 cases, and increased to 29 cases after treatment, of which 2 cases had HBV surface antigen negative. The HBV DNA content was between 1.0×103 and 1.0×106 before treatment in 13 cases, and after treatment in 3 cases, and the HBV DNA content was above 1.0×106 before treatment in 1 case, and after treatment in 1.6×104. However, in the combined TACE combined with RFA treatment group, the blood HBV DNA content in 1 case decreased below 1.0×103 from 1.1×105 before treatment. The difference of HBV-DNA content between the two groups It significantly reduces the content of hepatitis B virus in vivo or eliminates hepatitis B virus, and blocks the carcinogenic factors, but it needs further observation to improve the tumor-free survival of primary liver cancer.

Objective to investigate the regularity of radiation-induced Egr-1 gene promoter regulating adenovirus-mediated expression of Smad7 gene in mice lungs, its intracellular localization and its tolerance in mice. Methods the radiosensitive element of Egr-1 gene promoter and Smad7 cDNA were packaged into replication-defective adenovirus to prepare AD. Egr-Smad 7.120 C57BL/6J mice were randomly divided into the experiment, 6 mice in each group. The mice were administered intratracheally, and a single total thoracic radiation was administered 24 h later. (1) 66 mice were divided into 11 groups: saline group, AD group. Egr-Smad7 and unpackaged Egr-Smad7 adenovirus (each divided into 5 different virus titers) groups, the dyspnea, cyanosis and death within 72 h after administration were observed, and the relationship between the titers of recombinant adenovirus and acute tolerance of mice was investigated. (2) 36 mice were divided into 6 groups: blank control (C) group, radiation alone (R) group, AD. Egr-Smad7 non-radiation (RA) group, AD. (3) 168 rats were divided into 28 groups: C group, R group, RA group, RAR group, AV group and AVR group (each group was divided into 1×108, 5×108, 1×109 pfu/0.1 ml groups according to the adenovirus titer, and 6 h and 9 h groups according to the post-irradiation time). (4) 324 rats were divided into 54 groups: C group, R group, RA group, RAR group, AV group and AVR group (each group was divided into 0, 1, 2, 3, 5, 7, 9, 12 and 15 h groups according to the post-irradiation time). (5) 126 rats were divided into 21 groups: C group, R group, RA group, AV group and AVR group The relationship between the expression of Smad7 gene in lung and radiation dose was studied 5 h after irradiation.The expression of Smad7 protein in lung tissue of mice was detected by Western blot method.Results When the recombinant adenovirus titer was 109pfu or below, the mice could safely tolerate.All 6 mice in each group survived, while more than 5 mice in the group of 5×109 pfu or above died. The expression of exogenous Smad7 gene in AVR group was significantly expressed in the cytoplasm of alveolar epithelial cells, vascular endothelial cells and bronchiolar epithelial cells, but not in each control group. Its expression increased with the increase of radiation dose between 0 and 12 Gy and decreased at 15 Gy; Conclusion Mice can safely tolerate the expression of recombinant adenovirus at 109pfu or below, and the expression of exogenous Smad7 gene can be regulated by radiation-induced Egr-1 promoter in the cytoplasm of various cells in lung tissues of mice. The expression of exogenous Smad7 gene increases with the increase of recombinant virus titer, which is related to the dose and the interval time after radiation.

Objective to observe the effects of cimetidine on the growth, apoptosis, adhesion and invasion of human gastric cancer SGC-7901 cells. Methods the effects of different concentrations of cimetidine on the growth of gastric cancer SGC-7901 cells were detected by tetramethylazo salt (MTT) assay. Cell cycle and apoptosis were detected by flow cytometry; The morphological changes of the cells were observed by fluorescence microscope after acridine orange (AO) staining. The ultrastructural changes of cells were observed by transmission electron microscopy. The effect of cimetidine on the adhesion of gastric carcinoma cells to extracellular matrix was detected in matrigel. The effect of cimetidine on the ability of gastric carcinoma cells to invade artificial basement membrane was detected by millicell chamber. Results Cimetidine inhibited the growth of SGC-7901 significantly at the concentration of 0.5~5 mmol/L, and it was time and dose dependent; After 0.5~10 mmol/L cimetidine, the morphological changes of typical apoptosis were observed; The apoptosis peak was observed by flow cytometry, and the number of cells in Go/G1 phase increased significantly; Cimetidine 0.0625~0.25 mmol/L can inhibit the adhesion of gastric cancer cells to extracellular matrix and the invasion of recombinant basement membrane. Conclusion Cimetidine can change the cell cycle distribution, induce apoptosis of SGC-7901 cells, and thus inhibit cell proliferation.

Stem cells have the ability of multidirectional differentiation to produce different tissue cells, which can be induced to differentiate into cardiomyocytes under certain conditions, which is of great significance to the clinical treatment of myocardial infarction and cardiovascular tissue engineering, especially the research of cardiomyocardial tissue engineering.

Published April 2006<A study published in American Journal of Clinical Nutrition (Am J Clin Nutr, 2006, 83:880-860) showed that early age-related macular degeneration (AMD) is associated with a high glycemic index (GI) diet. AMD is one of the main causes of irreversible visual loss in the American population. The incidence rate is estimated to be 1.47% in people ≥40 years old, with 1.75 million patients. The incidence rate increases with age and is expected to increase by 50% in the next 20 years. Age-related macular degeneration (ARM) is an early manifestation of AMD, characterized by retinopathy pigmentosa.

mycoplasma penetrans (Mpe) is a newly discovered pathogenic mycoplasma associated with AIDS in recent years [1]. Tsais in 1986 and Zhang et al. in 1987 [2] reported that mycoplasma penetrans infected mouse embryonic cells at C3H for 6 weeks caused morphological changes of C3H cells without malignant characteristics, but malignant changes appeared at 11 weeks, and the transformation was irreversible after 18 weeks, and tumors formed in nude mice, suggesting that mycoplasma penetrans has carcinogenic potential [2]. In recent years, it has been further confirmed that mycoplasma infection is related to tumorigenesis [3], but there are few reports that mycoplasma penetrans has been isolated from tumor tissues and blood. In order to isolate and detect mycoplasma penetrans from blood and tumor tissues of tumor patients, we randomly selected 107 tumor patients diagnosed by pathological section from April 2004 to May 2005 in the First Affiliated Hospital of Wenzhou Medical College, Zhejiang Province. The tumor tissues and blood of mycoplasma penetrans were isolated and cultured, polymerase chain reaction (PCR), and sequence analysis. The blood and tumor tissue samples confirmed by sequence positive for Mycoplasma penetrans were taken to observe the existence of Mycoplasma penetrans particles and the ultrastructure changes of cells under electron microscope. The results are reported as follows.

Type 1 diabetes is characterized by β cell apoptosis, which is a T cell-mediated organ-specific autoimmune diabetes.To obtain sufficient number of islet cells for islet transplantation by islet stem cell technology and transgenic technology, it is necessary to deeply study the apoptosis mechanism of islet cells. Previous studies have shown that c-fos, transforming growth factor-β (TGF-β) and P21ras are related to apoptosis [1,2]. Therefore, we examined the expression of P21ras, TGF-β 1 and c-fos in human fetal fibroblasts and islet cells in incubation period, exponential growth phase and arrest phase, so as to further understand the apoptosis mechanism of islet cells.

tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor (TNF) family, which can effectively inhibit the formation and growth of many kinds of tumors and induce apoptosis of tumor cells, but has no toxic side effects on most normal cells [1,2]. Besides TRAIL ligand, death receptor antibody can specifically kill tumor cells like TRAIL, and its targeting ability to tumor cells is more superior than TRAIL, which has a good clinical application prospect. Guo et al. [3] reported a new mouse monoclonal antibody (called AD5-10) against the extracellular region of human TRAIL death receptor (DR) 5, which is a functional monoclonal antibody with great application value. The second mitochondria-derived activator of caspase (Smac) is a protein derived from mitochondria, which is released into the cytoplasm during apoptosis. High titers of recombinant viruses rAAV-Smac and rAAV-S3C were obtained after packaging. The anti-tumor activity of these two recombinant viruses and the clinical application prospect of their combination in tumor gene therapy were investigated in vitro and in vivo.

Case 1 male, 36 years old. Clinical diagnosis: coronary heart disease, old inferior myocardial infarction; Hypertension grade 2 (extremely high risk). The admission blood pressure was 150/85 mm Hg (1 mmHg =0.133 kPa), the heart rate was 87 beats/min, the rhythm was uniform. Coronary angiography showed diffuse right middle coronary lesion, 80% ~90% stenosis, 80% stenosis of middle anterior descending artery, distal myocardial bridge, left ventricular systolic ejection fraction (LVEF) was 65%. The patient was treated by surgical bypass. The right internal mammary artery and anterior descending artery were bypassed and the great saphenous vein and right coronary artery were bypassed by non-cardiopulmonary bypass and non-cardiac arrest. The ST segment of leads V1~ V4 of ECG showed an upward elevation of 0.4~0.6 mV, and the blood pressure was 80/60 mm Hg. Emergency coronary angiography assisted by intra-aortic balloon counterpulsation pump (IABP) was performed immediately. The anastomosis of internal mammary artery and anterior descending artery was occluded. The percutaneous transluminal coronary angioplasty (PTCA) with a diameter of 2.5 mm was used to dilate the anastomosis twice under 8~12 atm pressure. After the balloon catheter dilation, the internal mammary artery was recanalized without residual