Volume 87  ·   Issue 34  ·  2007

Professor Robert Steinbrook recently put forward some questions worth thinking about the formulation of clinical guidelines abroad [1]. In recent years, the guidelines in our country have more and more influenced the work of clinicians, and they should be referred to or borrowed from in almost every day work.

Objective to investigate the effects of tanshinone ⅡA, salvia miltiorrhiza, salvia polyphenolate and protocatechualdehyde on the proliferation of fibroblasts and the synthesis of procollagen type Ⅰ and Ⅲ in systemic scleroderma (SSc). Methods The effects of tanshinone ⅡA, salvia miltiorrhiza, salvia polyphenolate and protocatechualdehyde on the proliferation of fibroblasts were determined by microenzyme reaction colorimetry. The effects of tanshinone ⅡA, salvia miltiorrhiza, salvia polyphenolate and protocatechualdehyde on the mRNA synthesis of procollagen type Ⅰ and Ⅲ in fibroblasts were determined by reverse transcription-polymerase chain reaction. Results Tanshinone ⅡA, salvia miltiorrhiza, salvia polyphenolate and protocatechualdehyde all had inhibitory effects on the proliferation of fibroblasts in scleroderma (all P<0.01), and tanshinone ⅡA had the strongest effect. All four components had inhibitory effects on the expression of collagen type Ⅰ and Ⅲ mRNA in fibroblasts (all P<0.01), and tanshinone ⅡA was the strongest, which could reduce the mRNA of procollagen type Ⅰ Conclusion Tanshinone ⅡA has the strongest inhibitory effect on the proliferation of systemic scleroderma fibroblasts and the expression of type Ⅰ and Ⅲ procollagen mRNA, and has a strong anti-fibrotic effect.

Objective to investigate the detection and related characteristics of male genitourinary tumors in Beijing Hospital in recent 10 years.Methods the data of inpatients in urology department from January 1, 1995 to December 31, 2004 were investigated, and the newly diagnosed tumors were analyzed.Results the three most common malignant malignancies of male genitourinary system were bladder transitional epithelial cell carcinoma, prostate adenocarcinoma and renal clear cell carcinoma, with 387 cases, 271 cases and 250 cases detected respectively.The number of detected cases of these tumors showed a trend of increasing year by year.Among them, the early detection rate of prostate adenocarcinoma and renal clear cell carcinoma increased with the passage of time.In terms of the reasons for seeking medical treatment, prostate cancer changed the most, and the proportion of prostate cancer detected due to abnormal routine physical examination increased significantly in recent years.Conclusion The progress of various screening and diagnosis methods in recent 10 years has made the early detection of male genitourinary tumors changed significantly.

Objective to study the difference of gene expression profiles between patients with pulmonary embolism and normal controls, and to investigate the molecular mechanism of pulmonary embolism. Methods Fresh blood samples from 9 patients with pulmonary embolism and 33 normal controls were collected, total blood RNA was isolated, 9 cRNA probes labeled with Cy3 and 1 standard control probe labeled with Cy5 were synthesized, and hybridized with 44K human whole genome (Oligo) microarray, and verified by real-time fluorescence PCR. Results 434 genes were identified from 9 microarrays, including 36 genes involved in blood coagulation, 20 immune inflammation, 29 metabolism, 26 differentiation and apoptosis, 25 cell growth maintenance, 22 cell signal transmission, 14 cytoskeleton and motility, 15 ion channel and trafficking, 14 transcription, 6 RNA/DNA binding and others.

Objective to determine the frequencies of intrahepatic infiltrating lymphocytes (LILs) and immunoactive cells in peripheral blood of patients with chronic severe hepatitis B (chronic severe hepatitis B), and to compare and analyze their similarities and differences and their roles in the pathogenesis of chronic severe hepatitis B. Methods LILs from 11 patients with chronic severe hepatitis B who underwent liver transplantation were extracted and isolated by grinding with ground glass and natural sedimentation of hepatocytes. The frequencies of LILs and immunoactive cells in peripheral blood of patients with chronic severe hepatitis B were measured by flow cytometry in parallel and compared with those of normal subjects. Results (1) The frequencies of CD4+ T lymphocytes and B cells in LILs of patients with chronic severe hepatitis B were 17%±6% and 3.0%±1.0%, respectively, which were significantly lower than those in peripheral blood, which were 32%±8% and 21.4%±12.2%, respectively (all P<0.01), while the frequencies of CD8+ T lymphocytes, NK and NKT cells were 38%±13%, 34%±18% and 10%±4%, respectively, which were significantly higher than those in peripheral blood (2) Compared with the normal control, CD3+ and CD4+ T lymphocytes in LILs were significantly higher (P<0.05 or P<0.01), while NK cells and NKT cells were lower; (3) Compared with normal controls, the ratio of LILs to peripheral blood lymphocytes was higher in CD3+, CD4+, CD8+ T lymphocytes (CD4+ T lymphocytes P<0.01), and lower in B cells and NKT cells (P<0.01 or P<0.05). Conclusion A large number of immunoactive cells infiltrate in the liver, especially CD4+, CD8+ T lymphocytes and NK cells may play an important role in the pathogenesis of chronic severe liver.

Objective to evaluate the clinical efficacy and side effects of simultaneous accelerated intensity modulated radiotherapy (SMART) in newly treated nasopharyngeal carcinoma. Methods 110 cases of newly treated nasopharyngeal carcinoma were treated with SMART radiotherapy. The tumor and enlarged lymph nodes (GTV) were given 2.5 Gy/time for 28 times, with a total dose of 70Gy. Subclinical focuses and preventive irradiation areas received the conventional dose of 2.0 Gy/dose for 28 times, with a total dose of 56 Gy. Among them, 36 patients received GTV increased to 80 Gy. Results The median follow-up time was 24 months (7-44 months). The overall survival rate of 1, 2 and 3 years was 97.02%, 88.72% and 78.27%, respectively. The survival rate of 1 to 3 years without local recurrence was 97.94%. The survival rate of 1, 2 and 3 years without local recurrence and without distant metastasis was 95.21%, 89.83%, 76.10% and 95.38%, 85.71% and 79.67%, respectively. According to Fuzhou stage, the 3-year overall survival rate of stage Ⅰ to Ⅱ was 100%, stage Ⅲ was 74.33%, and stage IVa was 62.96%. Except for mucositis, the other acute toxic and side effects were well tolerated, and there was no grade 4 adverse reaction. The main toxic reaction in the later stage was dry mouth, and most patients showed grade 0 to 1 injury. There was no increase in toxic and side effects in patients with GTV reaching 80Gy. Conclusion SMART technology is effective for early

Objective to express human tyrosinase epitope peptide and to explore its application in the detection of autoantibodies in patients with vitiligo. Methods the target gene was synthesized and cloned into prokaryotic expression vector pGEX-4T-2, transformed into E. coli BL21 strain, and the protein expression was induced by isopropyl-β-D-thiogalactoside (IPTG). The protein expression was identified by sodium lauryl sulfate-polyacrylamide gel electrophoresis and Western blot. The protein was used as antigen to detect the serum IgG antibody in 100 patients with advanced vitiligo and 30 normal subjects. Results the recombinant expression vector was successfully constructed and the recombinant protein was successfully expressed. The expression of the protein was analyzed by gel analysis system, and it was found that the expression of the target protein accounted for 70% of the total protein in bacteria. After the fusion protein was purified by glutathione S-transferase kit, the protein purity was more than 90%. The serum binding reaction to the target protein was positive in 64 cases of 100 patients with vitiligo, and the serum binding reaction was negative in 30 normal subjects It is antigenic in the serum of patients with vitiligo.

Objective to compare the clinicopathological characteristics and prognosis of breast cancer at different stages in the past 20 years from 1981 to 2000, and to understand the trend of its incidence and prognosis. Methods 1678 cases of breast cancer were randomly selected from 11 400 cases admitted to the Affiliated Cancer Hospital of Tianjin Medical University in the past 20 years from 1981 to 2000, and their clinicopathological and prognostic data were retrospectively analyzed and compared. Results Compared with the 1990s and 1980s, the number of breast cancer cases admitted increased by 237%, with an average annual increase of 11.8%; There was no significant change in the onset age of patients, the median age was 46~47 years old; The detection rate of early stage breast cancer (stage 0~ Ⅰ) increased from 12% in 1981-1985 to 15.5% in 1996-2000, while the proportion of stage Ⅲ breast cancer decreased significantly from 24.9% to 16.5%, and the proportion of carcinoma in situ increased from 3.7% to 4.4%; At the same time, the rate of axillary lymph node metastasis and the chance of local recurrence and distant metastasis decreased from 51.8% and 27.8% to 46.3% and 13.5%, respectively; Survival analysis showed that the 5-year and 10-year survival rates increased from 76% and 63% in 1980s to 86% and 74% in 1990s, respectively. Conclusion In recent 20 years, the incidence of breast cancer showed an obvious upward trend, the detection rate of early breast cancer increased, the rate of axillary lymph node metastasis and the probability of local recurrence and distant metastasis decreased, the prognosis of patients improved, and the 5-year and 10-year survival rates increased.

Objective to establish a real-time quantitative PCR method for the detection of JAK2V617F mutation in patients with myeloproliferative diseases (MPD). Methods TaqMan-MGB probe was used to detect JAK2V617F mutation in patients with MPD. Some specimens were verified by allele-specific qualitative PCR and sequencing simultaneously. Results 374 bone marrow or peripheral blood specimens from patients and controls were used for the analysis of JAK2V617F mutation. Only 1 of 65 cases of acute myeloid leukemia was positive, while the detection rate in bone marrow cells of 38 cases of chronic myeloid leukemia, 30 cases of acute lymphocytic leukemia, 8 cases of chronic lymphocytic leukemia, 7 cases of non-Hodgkin's lymphoma and 16 cases of normal donors was 0. Conclusion JAK2V617F mutation is widespread in MPD patients in China. Real-time quantitative PCR with TaqMan-MGB probe can detect JAK2V617F mutation rapidly and accurately.

Objective to investigate whether angiogenesis mimicry (VM) exists in malignant tumors of digestive tract with bidirectional differentiation. Methods the angiogenesis pattern of malignant tumors of digestive tract with bidirectional differentiation was studied by immunohistochemistry and PAS staining. 111 cases of malignant tumors of digestive tract with bidirectional differentiation were collected, including 80 cases of malignant gastrointestinal stromal tumors; 18 cases of malignant melanoma; 13 cases of carcinosarcoma were stained with vascular endothelial growth factor (VEGF), CD31 and PAS, the expression of VEGF was calculated by stereological grid counting method, and the duct area surrounded by CD31 and PAS positive patterns, namely, microvessel density (MVD) and VM density (VMD), were calculated under microscopic image analyzer, respectively. VM was studied according to the expression levels of the three, and the relationship between the three and the malignant degree of tumor was compared and analyzed. Results VM was formed in malignant tumors with bidirectional differentiation in digestive tract, and red blood cells were present in it. The expression of VEGF (89 ± 20) and MVD (47 ± 12) in tumors containing VM were lower than those without VM (126 ± 18, 78 ± 13, all P<0.05); With the increase of malignancy, the proportion of VM tumors gradually increased, and the expression of VEGF, MVD and VMD in VM tumors gradually increased. The above indexes were significantly different between low-grade malignancy (45±19, 15±8, 38±25) and high-grade malignancy (128±42, 81±17, 122±39) (all P<0.05). Conclusion the malignant cells with bidirectional differentiation in digestive tract have plasticity, and VM exists.

Objective to investigate the relationship between the functional status of peripheral blood mononuclear/macrophage marker antigen CD14 and its early activating molecule CD69 in HIV/AIDS patients in China, and to investigate the relationship between the functional status of peripheral blood mononuclear/macrophage and the disease progression of HIV infected patients. Methods Anticoagulated whole blood of 57 HIV/AIDS patients without antiviral treatment and 32 healthy people was collected. The expression of CD4+ T lymphocytes, CD14+ cells and CD69 antigen was determined by flow cytometry, and the plasma HIV-1 load was detected by real-time fluorescence quantitative RT-PCR. Results (1) The particle size SSC value of CD14+ cells in peripheral blood of HIV/AIDS patients was 76±16, and the expression rate of CD69 molecule in CD14+ cells was 27%±4%, which was significantly higher than that of the control group (P<0.05). In AIDS group, the expression rate of CD14+ cells was higher than that of the chronic HIV infection (HIV) group and the long-term non-progression (LTNP) group, and the expression rate of LTNP in HIV group was higher than that of the control group (P<0.05). There was no significant difference in the proportion of CD14+ cells and CD14 antigen level in peripheral blood compared with the control group. (2) The expression rate of CD69 molecule in peripheral blood CD14+ cells in HIV/AIDS patients was negatively correlated with the absolute value of CD4+ T lymphocytes (r = -0.872, P<0.01), and positively correlated with HIV-1 RNA viral load (r =0.697, P<0.01). Conclusion The activation and phagocytosis of peripheral blood mononuclear/macrophages in HIV/AIDS patients in some areas of China were significantly higher than those in healthy controls, and they were closely related to the disease progression.

Objective to investigate the effects of aldosterone and its receptor blocking on the production of plasminogen activator inhibitor 1 (PAI-1) in mesangial cells. Methods the changes of PAI-1 mRNA in mesangial cells were observed by RT-PCR after aldosterone alone was stimulated for 24h and after blocking with spironolactone. Western blot was used to detect PAI-1 in cell supernatant, and ELISA was used to determine the content of transforming growth factor β 1 (TGF-β 1) in cell supernatant; The level of reactive oxygen species (ROS) in mesangial cells was detected by confocal laser. The effects of different concentrations and different action times of aldosterone on PAI-1 mRNA in mesangial cells were observed. Results Aldosterone significantly increased the expression of PAI-1 mRNA and protein in mesangial cells, as well as the expression of TGF-β 1 and the level of ROS in cells. Aldosterone increased the expression of PAI-1 mRNA in a dose-dependent manner. The expression of PAI-1 mRNA increased significantly after 100 nmol/L aldosterone stimulated mesangial cells for 4 h. After adding spironolactone, the increased expression of PAI-1, TGF-β 1 and intracellular ROS returned to normal levels. Conclusion Aldosterone can independently promote the increase of PAI-1 secretion in rat mesangial cells, and at the same time, aldosterone increased the expression of TGF-β 1 and intracellular ROS levels, which were mediated by mineralocorticoid receptor.

Objective to study the biocompatibility of extracellular matrix (ECM) of allogeneic fascia lata and to explore the development of a new material for repairing renal injury. Methods Dog fascia lata was treated with acellular fluid, DNase and RNase to prepare allogeneic fascia lata ECM. The extract of fascia lata ECM was prepared and tested for acute systemic toxicity in Kunming mice; The cytotoxicity of mouse fibroblasts was detected by tetramethylazolazole salt (MTT) colorimetric assay. The left kidney of dogs was made into a renal injury model and repaired with allogeneic fascia lata ECM. The repaired local samples were examined by light microscope and scanning electron microscope at different periods after operation. Results The mice had no abnormal reaction 4 hours after injection of fascia lata ECM extract. The body weight increased from day 1 to day 3, and there was no death or poisoning reaction, which was not significantly different from the negative control saline group. The relative proliferation rate (RGR) of the mice was 112% (day 2), 96% (day 4) and 97% (day 7) by MTT assay, and the toxicity assessment was 0, 1 and 1. The material had no obvious cytotoxicity. There was no obvious inflammatory cell infiltration and no obvious adhesion to the surrounding tissues in the local area repaired by ECM material at all periods after operation. Conclusion Allogeneic fascia lata ECM has good biocompatibility and is expected to be an ideal repair material for renal injury.

Objective to investigate the tolerance of PC12 cells to ischemia induced by ginkgolide (Gin) pretreatment and its related mechanism.Methods Rat adrenal pheochromocytoma PC12 cells were randomly divided into control group,ischemic 9 h group,ischemic preconditioning (IP) 1.5 h + ischemic 9 h group,and ginkgolide preconditioning + ischemic 9 h group.The effects of ginkgolide preconditioning on PC12 cell ischemia model were compared by thiazole blue colorimetry,cell morphology,Western blot and gel electrophoresis migration experiment.Results In the PC12 cell ischemia model, 9 h ischemia significantly reduced the viability of PC12 cells, and the cell survival rate was (49.3±2.8) %, while 24 h ginkgolide preconditioning significantly improved the survival rate of 9 h ischemic cells (65.9±2.8) % (P<0.01); The cell morphology showed that the cell damage caused by ischemia was significantly reduced after ginkgolide pretreatment for 9 h. More cells retained processes, the network of processes still existed, and the cell body was intact. Compared with the control group, ginkgolide pretreatment could significantly increase the expression of hypoxia-inducible factor-1 α (HIF-1 α) protein (P<0.01); The expression of HIF-1 induced by ginkgolide pretreatment has biological activity, it can bind to DNA and activate the expression of the downstream gene erythropoietin (EPO) protein (P<0.01). Conclusion Ginkgolide pretreatment can induce the tolerance of PC12 cells to ischemia, and its induction of tolerance may be related to the stable expression of HIF-1 α in PC12 cells induced by ginkgolide pretreatment, the increase of the binding activity of HIF-1 to DNA and the increase of the expression of the downstream gene EPO protein.

Objective to identify the specific binding protein of β 2 glycoprotein Ⅰ (β 2gpⅠ) on the cell membrane of hepatocellular carcinoma cell line SMMC-7721 and its function. Methods A self-made β 2gpⅠ affinity column was used to amplify annexin Ⅱ from hepatocellular cell line SMMC-7721 by reverse transcription PCR. The competitive binding of annexin Ⅱ and β 2gpⅠ-GFP to SMMC-7721 was detected by flow cytometry. The results showed that the cDNA fragment of β 2gpⅠ specific binding protein was about 1.1kb, which was highly homologous to human annexin Ⅱ (98%). Annexin Ⅱ was expressed in SMMC-7721, and β 2gpⅠ-GFP and SMMC-7721 in 1 μ L and 4 μ L of annexin Ⅱ could be combined with SMMC-7721 respectively

The concept of metabolomics was first put forward by Nicholson et al. in 1999. Its significance is to study the dynamic changes of metabolites caused by pathophysiological stimulation and some genetic modification of organisms, so as to obtain information about the changes of metabolites in organisms with time and biochemical processes [1].

Total cholesterol, low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) have been confirmed as independent predictors of cardiovascular disease, but the importance of triglycerides is still controversial.

Many studies have shown that organic foods contain anti-tumor substances. A systematic review has found that eating more fruits and vegetables can reduce the risk of breast cancer, while eating more fat can increase the risk of breast cancer.

In recent ten years, there has been a lot of concern about statistical errors or defects in published medical papers in China [1]. Although the classification of errors or defects is different, many researchers have mentioned the problems of statistical expression or reporting [2-3].