Volume 86  ·   Issue 02  ·  2006

Colorectal cancer is a digestive tract tumor that can be prevented and treated, and it is one of the best models in many tumor studies [1]. It is of great clinical and social significance to strengthen the clinical application research of colorectal cancer, including whether the intestine is prepared before operation, the advantages and disadvantages of laparoscopic application, the follow-up procedure and the necessity of follow-up, etc.

Objective to evaluate the effect of sympathetic nerve stimulation on ventricular repolarization in different regions of the heart after myocardial infarction. Methods Fourteen dogs were randomly divided into myocardial infarction group (7 dogs) and sham operation control group (7 dogs). In the myocardial infarction group, myocardial infarction model of left ventricular anterior wall was caused by surgical ligation of the main anterior descending coronary artery. The effective refractory period (ERP) of the proximal heart base and distal apex adjacent to the infarction area was measured by program 4 weeks after operation. Results In the myocardial infarction group without left stellate ganglion stimulation, there was no significant difference between the ERP value (162 ms ± 9 ms) at the proximal base of the infarction area and the ERP value (161 ms ± 6 ms) at the distal apex of the infarction area (P>0.05), which was similar to that in the sham-operated control group (162 ms ± 10 ms vs 164 ms ± 5 ms, P>0.05); However, after sympathetic nerve stimulation, only the ERP value at the apex of myocardial infarction group showed no obvious change, and the ERP value in other measured areas showed shortening phenomenon. In myocardial infarction group, ventricular repolarization was dispersed between the base of heart (141 ms ± 10 ms) and the apex of heart (157 ms ± 8 ms) (P<0.05). In myocardial infarction group, the sympathetic nerve immunohistochemical staining at the site where the apex electrode was located was negative, and the normal immunohistochemical distribution of nerve fibers was observed at the base of heart and the sham operation control group. Conclusion There is regional denervation in the heart after myocardial infarction; Abnormal distribution of autonomic nerves leads to the occurrence of ventricular repolarization dispersion after nerve stimulation.

Objective to investigate the expression of normal RhD protein in individuals with Del blood group. Methods RT-PCR and cDNA sequence analysis were used to detect RhD mRNA in individuals with different Rh phenotypes of Del blood group (ccdeldeldelee, CcDeldee and CCDelDelee) and analyze the expression of Del protein. Results There were complex RhD mRNA forms in individuals with Del blood group. Six forms of RhD transcripts were detected, including deletion of exons 7 to 9 (Del789), deletion of exons 7 and 9 (Del79), deletion of exons 8 and 9 (Del89), deletion of exon 9 (Del9), deletion of exons 8 and 9 plus 170 bases of intron 7 (Del89+170) (917-1086, Genbank AB035194), deletion of exon 9 plus 170 bases of intron 7 (Del9+170). All of these transcripts were deletion of exon 9, and transcript Del9 was the most similar to normal RhD mRNA. Conclusion There is no normal RhD mRNA in Del blood group.

Objective to analyze the gene of coagulation factor VII (FVII) in a patient with hereditary coagulation factor VII (FVII) deficiency and its family members, and to reveal the molecular mechanism of the pathogenesis of FVII. Methods Genomic DNA of peripheral blood of proband and his family members was extracted, and 8 exons and flanking sequences of FVII gene were amplified by polymerase chain reaction (PCR), and the abnormalities of FVII gene were detected by nucleotide sequence analysis; The PCR products of proband mutant sequences, family members and 100 normal human corresponding sequences were digested with restriction enzyme ECO91I to further identify the gene mutation sites and exclude gene polymorphism; The molecular structure and pathology of the gene mutation were analyzed by using protein molecular model simulation software.Results A → G heterozygous mutation occurred in nucleotide 10833 of exon 8 of proband FVII gene, resulting in Met306Val; C→A heterozygous mutation occurred in nucleotide 9643 of exon 7 of proband, resulting in Thr181Asn. The former was inherited from the mother and the latter was inherited from the father, and the proband's sister was heterozygous for A10833G (Met306Val). The spatial configuration simulation analysis of protein found that Met306Val mutation was located on the surface of FVII molecule, which produced steric hindrance and affected the structure and function of the protein. Conclusion The double heterozygous mutation of Met306Val and Thr181Asn in FVII gene is the molecular mechanism leading to this hereditary coagulation factor VII deficiency; Met306Val mutation is a new type of mutation that has not been reported in the world. It is suggested that the spatial configuration of FVII protein can be changed by the mutation of Met306Val.

Objective to compare the efficacy and adverse reactions of amiodarone and sotalol on atrial fibrillation (AF). Methods 102 patients with atrial fibrillation were randomly divided into amiodarone group (51 cases) and sotalol group (51 cases). The amiodarone group was given amiodarone 600 mg/d, 7 days, 400 mg/d for 7 days, then reduced to 200 mg/d, and maintained at 200 mg/d after converting to sinus rhythm. The sotalol group was given sotalol 40~80 mg/d, 7 days, increased to 160 mg/d in the second week, and maintained at 40~80 mg/d after converting to sinus rhythm. The patients in both groups were stopped after 3 weeks of converting sinus rhythm. Follow-up for 12~24 months, and the efficacy was judged by echocardiography, electrocardiogram and 24-h dynamic electrocardiogram before and after treatment. Results (1) 40 cases of atrial fibrillation successfully cardioverted in amiodarone group, with an effective rate of 78.4%; 36 cases of atrial fibrillation in the sotalol group were successfully cardioverted, with an effective rate of 70.6%. (2) 34 cases in the amiodarone group and 10 cases in the sotalol group were converted to sinus rhythm within 1 week. (3) After 12 months follow-up, 27 cases (67.5%) in the amiodarone group and 15 cases (41.7%) in the sotalol group still maintained sinus rhythm; After 24 months, 12 patients (44.4%) in the amiodarone group maintained sinus rhythm, and 4 patients (26.7%) in the sotalol group. (4) After 6-12 months follow-up, 10 patients in the sotalol group (80 mg/d) stopped taking the drug due to atrioventricular block and severe bradycardia, but no serious arrhythmia occurred in the amiodarone group. (5) The duration of atrial fibrillation>12 months is a predictive factor for the difficulty of maintaining sinus rhythm.

Objective to investigate the clinical value of contrast-enhanced ultrasound-guided percutaneous puncture biopsy of liver tumors. Methods 211 cases of liver space-occupying lesions of unknown nature were randomly divided into two groups in the Department of Ultrasound, Peking University School of Clinical Oncology. Among them, 96 cases were treated with conventional ultrasound-guided puncture biopsy after 149 foci were treated with contrast-enhanced ultrasound agent SonoVue (CEUS group), and 115 cases with 153 foci were treated with ultrasound-guided puncture biopsy without contrast-enhanced ultrasound (US group). There was no significant difference in the size distribution of lesions between the two groups (P>0.05). Among them, 67 cases (24.8%) of micromalignant tumors ≤2.0 cm, 12.9% in CEUS group and 11.9% in US group, respectively. The results of pathological diagnosis of biopsy were combined with surgery, enhanced CT, magnetic resonance imaging (MRI), angiography, alpha-fetoprotein (AFP) examination or clinical follow-up for 3~6 months, and the results of puncture biopsy were analyzed and compared between the two groups. Results The number of satisfactory specimens obtained after ≤2 punctures in CEUS group was significantly more than that in US group (P =0.0007). The successful rate of 149 lesions in CEUS group was 98.7% (n =147), and the confirmed rate was 96.0% (n =143). The diagnosis rate of small malignant tumors ≤2.0 cm was significantly higher in the CEUS group (97.4%) than in the US group (80.6%, P =0.0473). In addition, 15 (13.4%) of 112 malignant tumors in the US group were negative by puncture biopsy, and 14 were confirmed as malignant by contrast-enhanced ultrasound guided biopsy, of which 7 were ≤2.0 cm. There was no complication in the US group except 1 case of pneumothorax. Conclusion Contrast-enhanced ultrasound guided puncture biopsy is helpful to judge malignant lesions and small tumors, and confirm necrosis and liquefaction areas, which makes the location and sampling more accurate than conventional ultrasound, thus greatly improving the success rate of percutaneous puncture biopsy and the diagnosis rate of malignant lesions, and significantly reducing the false negative rate of puncture biopsy, which has important clinical application value.

Objective to compare and analyze the effects of cyclosporine and tacrolimus (FK506) on the replication of hepatitis B virus (HBV) in vitro. Methods HBV DNA was permanently transfected into human hepatoma cell line HepG2.2.15 as an in vitro experimental model. After the safe concentrations of cyclosporine and FK506 were determined by tetramethylazolazole blue (MTT) assay, different safe concentrations of cyclosporine (0.6~20.0 μ g/ml) or FK506 (5~100 ng/ml) were applied to the cell line, and the corresponding concentration of fresh drug-containing medium was changed every 24 hours. After the drug was applied for 4 days, the culture supernatant and cultured cells were collected and tested. The levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the supernatant were detected by enzyme-linked immunosorbent assay, and the intracellular HBV DNA levels were semi-quantitatively analyzed by slit blot hybridization, so as to comprehensively evaluate the effects of cyclosporine and FK506 on the replication of HBV virus Within the safe concentration range, with the increase of cyclosporine concentration, the inhibition rate of HBsAg and HBeAg increased gradually, and the intracellular HBV DNA replication level decreased; When 1.3, 2.5 and 5.0 μ g/ml cyclosporine were treated for 4 days, the inhibitory rates of HBsAg were 16.5%±9.4%, 21.5%±8.9% and 33.1%±5.3%, respectively, and the inhibitory rates of HBeAg were 7.8%±2.2%, 11.0%±2.3% and 20.8%±1.5%, respectively. The expression of HBV DNA was only 56.1%±16.5%, 41.7%±10.9% and 39.5%±9.5% of that of the control group. Within the safe concentration range, FK506 had no obvious inhibitory effect on the expression and secretion of HBsAg and HBeAg, and intracellular HBV DNA replication in HepG2.2.15 cells. However, FK506 does not have this effect.

Objective to construct a shuttle plasmid for the fusion gene of herpes simplex virus thymidine kinase (HSV-tk) and enhanced green fluorescent protein (EGFP) driven by human insulin-like growth factor Ⅱ gene (IGF-Ⅱ) P3 promoter, and to observe its expression and effect in hepatoma cell line HepG2 and cervical cancer cell line HeLa cells.Methods The shuttle plasmid vector for the fusion expression of EGFP and HSV-tk driven by IGF-ⅡP3 promoter was constructed by gene recombination technique.The shuttle plasmid vector was transfected into HepG2 and HeLa by liposome transfection technique.After 48 h, the fluorescence expression was observed under fluorescence microscope.The mRNA expression of HSV-tk and EGFP was detected by reverse transcription polymerase chain reaction (RT-PCR), and the cytotoxicity of ganciclovir at different concentrations on HepG2 and HeLa cells after transfection with the fusion protein vector was detected by tetramethylazo blue assay.Results The recombinant plasmid was successfully constructed by enzyme digestion and sequencing analysis. Among the cells transfected with this shuttle plasmid, only HepG2 cells detected green fluorescence; RT-PCR detected that the mRNA of HSV-tk and EGFP were expressed only in HepG2 cells; Gancyclovir (GCV) had a selective cytotoxic effect on HepG2 cells transfected with this plasmid vector, and the inhibition rates of GCV at 3 concentrations of 1, 10, and 100 μ g/ml for 72 h (all experiments were repeated three times) were 19.8%±1.3%, 36.2%±2.0% and 48.7%±1.9%, respectively, compared with those of HepG2 cells transfected with this plasmid group (24.1%±1.9%, 45.1%±1.7%, 69.4%±3.6%) and HeLa cells transfected with this plasmid group (25.1%±1.6%, 49.3%±1.1%, 72.2%±2.9%), the differences were statistically significant (all P<0.01). Conclusion The shuttle plasmid vector driven by IGF-ⅡP3 promoter carrying the fusion gene of HSV-HSV-tk and EGFP was successfully constructed, and it can selectively kill the human hepatoma cell line HepG2 after transfection, which lays a foundation for further targeted adenovirus gene therapy of hepatoma.

Objective to confirm the differentiation and proliferation of cardiomyocytes in adult rats with heart failure, and to investigate the correlation between the degree of cardiomyocyte proliferation and cardiac function.Methods Adult SD male rats were divided into two groups: myocardial infarction group (14 rats, including 8 with compensated cardiac function and 6 with decompensated cardiac function) and normal control group (8 rats). Myocardial infarction was formed by ligation of the anterior descending branch of left coronary artery, and the rat model of heart failure was established.After 1 month, hemodynamics was measurement.Myocardial tissues were stained with propidium iodide and α-myoganglion actin antibody immunofluorescence to observe the mitotic phase of cardiomyocytes, and immunohistochemical staining with proliferating cell nuclear antigen (PCNA) antibody was performed to observe the positive rate of PCNA. Results (1) The mitotic phase of cardiomyocytes was observed in left ventricular myocardial tissues of adult rats. (2) The positive rate of PCNA in the cardiac function compensated group after myocardial infarction was significantly higher than that in the control group (7.2%±1.4%vs 2.2%±0.8%,P<0.01), but there was no significant difference compared with that in the cardiac function decompensated group (3.0%±1.3%vs 2.2% The positive rate of PCNA and the correlation coefficient of ±LVdp/dtmax in the myocardial infarction group were 0.80 (P<0.01) and-0.76 (P<0.01), respectively. Conclusion (1) There is cardiomyocyte proliferation in the heart of adult heart failure rats; (2) The proliferation degree of myocardial cells in rats with myocardial infarction was positively correlated with systolic function and negatively correlated with diastolic function.

Objective to investigate the effects of p53 inhibitor PFT-α (PFT-α) on the proliferation and cycle of colonic epithelial cells and the mechanism of cycle regulation. To observe the effects of PFT-α on the proliferation of colonic epithelial cells injured by thermochemotherapy. Methods the effects of different concentrations of PFT-α on the proliferation of colonic epithelial cells were observed by tetramethylazolazole blue (MTT) assay. After adding different concentrations of PFT-α to primary cultured colonic epithelial cells for 30 min, annexin V-fluorescein isothiocyanate (FITC) /propidium iodide (PI) staining, cell apoptosis was detected by flow cytometry, and cell cycle was detected by PI staining. The expression of cyclin B1 and Cdc2 (Tyr15) in colonic epithelial cells was detected by Western blot, and the expression of cyclin B1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Results PFT-α had no effect on the proliferation of colonic epithelial cells when it was lower than 50 μ mol/L, but had an inhibitory effect on the proliferation of colonic epithelial cells when it was higher than 60 μ mol/L Cell cycle analysis by flow cytometry showed that G2/M of colonic epithelial cells was prolonged after PFT-α, and the expression of cyclinB1 and Cdc2 (Tyr15) protein increased gradually with the increase of PFT-α dose.Conclusion PFT-α may promote the expression of cyclinB1 protein and mRNA, increase the phosphorylation level of Cdc2 (Tyr15), decrease the activity of cyclinB1/Cdc2, and reduce the damage of colonic epithelial cells by thermochemotherapy.

Objective to investigate the effect of preoperative hepatic artery combined with regional arterial infusion chemotherapy (PHRAIC) on preventing liver metastasis after radical resection of colorectal cancer. Methods 222 patients with colorectal cancer were randomly divided into PHRAIC group (110 cases) and control group (112 cases). There were no significant differences in sex, age, tumor location, tumor size, stage, tumor differentiation degree, postoperative chemotherapy and follow-up time between the two groups. The hepatic artery combined with regional arterial infusion chemotherapy (FUDR) + mitomycin (MMC) + oxaliplatin) was treated 7 days before operation in PHRAIC group, while the control group was treated directly with surgery. Both groups received routine systemic intravenous chemotherapy after operation. The liver metastasis rate, tumor-free survival rate, overall survival rate and median survival rate were observed 3 years after follow-up. Results There were no significant differences in postoperative complications between PHRAIC group and control group. The 3-year liver metastasis rate, 3-year tumor-free survival rate, overall survival rate and median survival rate of PHRAIC group and control group were 12.7% and 28.3%, respectively (P =0.01) Conclusion Preoperative hepatic artery combined with regional arterial perfusion chemotherapy is safe and effective, and can significantly reduce the incidence of postoperative liver metastasis and prolong the survival of patients with stage Ⅲ colorectal cancer.

Objective to investigate the regulatory effects of nuclear factor E2p45-related factor 2 (Nrf2) gene on the expression of uridine diphosphate glucuronyltransferase (UGT) 1A and its isoenzymes UGT1A8 and UGT1A10 in colon cancer cells induced by epigallocatechin gallate (EGCG). Methods EGCG was treated with Caco-2 and HT-29 colon cancer cells for 12 h, and the changes of gene expression were detected by reverse transcription polymerase chain reaction (RT-PCR). The changes of Nrf2 protein localization were observed by immunocytochemistry and laser confocal microscopy. In addition, the expression of Nrf2 gene was inhibited by RNA interference technique, and the effects of EGCG on gene expression were observed before and after blockade. Results (1) There were differences in the expression levels of UGT1A, 1A8 and 1A10 in Caco-2 and HT-29 colon cancer cell lines, colon cancer and adjacent tissues. (2) Nrf2, UGT1 were observed after EGCG was treated with Caco-2 and HT-29 cells (4) pSilence-Nrf2-B plasmid could significantly inhibit the expression of Nrf2 gene in Caco-2 cells and HT-29 cells with inhibition rates of 81.46%±1.68% and 84.72%±2.08%, respectively (the experiment was repeated three times); Caco-2-siNrf2 and HT-29-siNrf2 cell lines with low expression of Nrf2 were stably screened and established. Not only the expression level of basal UGT1A enzyme was reduced, but also the effect of enzyme-induced expression disappeared after EGCG. Conclusion Nrf2 gene participates in the process of EGCG-induced UGT1A and its isoenzyme expression and plays a key role in it, which provides a new argument for further demonstrating the role of EGCG in chemoprophylaxis of colon cancer and its possible mechanism.

Objective to investigate the relationship between cell signaling and transcription activator 6 (STAT) signaling pathway and apoptosis of colon cancer cells. Methods Four STAT6-specific shRNA plasmid vectors were constructed and transiently transfected with cationic liposomes in human colon cancer cell line HT-29 to block the STAT6 signaling pathway. The expression of STAT6 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) and the expression of phosphorylated STAT6 protein (pSTAT6) was detected by flow cytometry to screen effective small hairpin RNA (shRNA). The changes of apoptosis were analyzed by flow cytometry; The expression of Bcl-2 and Bax gene was observed by RT-PCR and Western blot. Results Two shRNA vectors specific for STAT6 were successfully constructed and screened. The apoptosis rate of STAT6 gene-silenced colon cancer cells increased significantly, and the early apoptosis rate was 0.4% in blank control group, 13.0% in pshRNA-STAT6-1 group and 8.8% in pshRNA-STAT6-4 group, respectively (P<0.05); The expression of Bcl-2 gene was significantly reduced, but the expression of Bax protein was significantly increased in colon cancer cells with Stat6 signaling pathway blocked (P<0.01). Conclusion Stat6 signaling pathway can inhibit apoptosis of colon cancer cells, probably by regulating the expression of Bcl-2 and Bax gene.

A 37-year-old male was found to have a mass in the anterior chest for more than 30 years, which increased slowly and gradually, and now was about the size of a goose egg.

Complications such as infection, excessive keratosis of nail bed and new nail deformity are often associated with peeling and loss of nail, exposed nail bed, and sharp local pain. When repairing nail bed injury, the shedded nail plate is often replanted, but many clinical patients have lost nail plate and cannot be replanted. From February 2002 to December 2004, on the basis of measuring the width and thickness of healthy adults' nail (toe), the Department of Orthopaedics, Zhaoyuan Hospital of Traditional Chinese Medicine, Shandong Province, treated the root of the orthopaedic feather shaft of domestic geese to make a replacement nail plate, and glued it to the nail bed with medical glue to treat the nail bed injury, with satisfactory curative effect. The report is as follows.

Thymoma is the most common tumor of anterior mediastinum, accounting for 83% ~90% of anterior mediastinal tumors and 20% of mediastinal tumors. Thymoma complicated with simple erythrocyte aplasia anemia (PRCA) is a relatively rare disease. The data of 3 patients with thymoma complicated with PRCA who underwent surgical treatment in the Department of Oncology of the Second Affiliated Hospital of Zhejiang University School of Medicine from 2000 to 2005 were analyzed and summarized.

A 40-year-old male with a long history of hypertension suffered from severe pain in the chest and back in 2001, but the symptoms disappeared after symptomatic treatment.

Scientists in the United States have recently said that the gene of the 1918 "Spanish flu" virus reveals the evolutionary trend of avian influenza virus. The information gained by scientists will help people predict the evolutionary direction of H5N1 avian influenza and know when this virus can cause a major human epidemic in time.

According to statistics, among 1415 known human pathogens, 61% belong to animal origin; Among the pathogens of new infectious diseases, 75% are zoonotic (can be transmitted from animals to humans).

A 50-year-old male was admitted to hospital for 8 months due to anorexia, fatigue, ascites and abdominal distension.Physical examination showed abdominal distension, multiple thoracic and abdominal varices, blood flow from bottom to up.The liver was 3 cm below the costal arch, 6 cm below the xiphoid process, and 3 cm below the costal arch of the spleen.

The first Beijing International Digestive Diseases Summit Forum, co-sponsored by the Digestive Diseases Research Center of Peking University School of Medicine, China Medical Forum Newspaper, Massachusetts Medical Association and New England Journal of Medicine, was held in Beijing from November 4 to 6, 2005.

At the winter meeting of the Society of Urologic Oncology (SUO) held in Bethesda, Maryland, USA from December 2 to 3, 2005, four scholars exchanged their research results on new methods of imaging diagnosis of urinary tract tumors.