Volume 86  ·   Issue 19  ·  2006

Objective to investigate the therapeutic effect and mechanism of brain tissue specific angiogenesis inhibitory factor 1 (BAI1) on glioblastoma. Methods BAI1-adenovirus vector was constructed by COS-TPC method. The successful construction of virus recombinant and transfection of tumor cells were verified by RT-PCR. Glioblastoma cell lines U87MG were inoculated into the brain of nude mice by stereotactic method. After tumor formation, the recombinant virus AdeBAI1 (AdeBAI1 group) or AdeLacZ (Mock) (AdeMock group) were injected into the tumor. Six nude mice in both groups were injected into the tumor. The survival time of nude mice was observed. Three glioblastoma cell lines SW1783, U87MG and U373MG were transfected with AdeBAI1 or AdeMock. The cells were collected 48 h later, and the viable cells were counted by MTF method. Total RNA was extracted with Trizol reagent. The mRNA expression of BAI1 and other angiogenesis-related factors was studied by RT-PCR. Results the mRNA expression of BAI1 was found only in AdeBAI1 transfected cells. The results of intracranial glioblastoma treatment showed that the mean production period was (26.0±4.6) d in AdeBAI1 group and (17.3±2.3) d in AdeBAI1 group, P<0.05. The cell count after transfection was (2.12±0.18) ×105 in AdeBAI1 group and (4.23±0.18) ×105 in AdeBAI1 group, P<0.05. This suggests that the proliferation of glioblastoma cells can be inhibited by BAI1. After AdeBAI1 transfection, the expression of angiogenesis-related factors Angiostatin and VEGF decreased, while the expression of VEGF-B and TSP1 increased. Conclusion Intratumoral injection of AdeBAI1 can inhibit the growth of glioblastoma. The increased expression of BAI1 can affect the expression of other angiogenesis-related factors. The increased expression of BAI1 can inhibit the proliferation of tumor cells. The antitumor effect of BAI1 may come from the inhibition of angiogenesis

Objective to investigate the relationship between single nucleotide polymorphism (SNP) of megsin gene and IgA nephropathy. Methods All coding regions, some regulatory regions and exon-intron junction regions of megsin gene were screened by direct sequencing. Eleven single nucleotide polymorphisms were found. Eight SNPs (frequency>0.05) were selected. 210 Chinese Han patients with sporadic primary IgA nephropathy were selected, and 103 healthy volunteers were used as the control group. The genotyping of IgA nephropathy was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing of PCR products. Results 267G/A of 5' untranslated region in exon 1 was significantly significant in the pathogenesis of IgA nephropathy: the frequency of AG + AA genotype in IgA nephropathy group was 29.0% (65/210), significantly higher than 16.5% (17/103), P<0.05; the frequency of allele A was 14.8% (62/210), significantly higher than 8.7% (18/103), P<0.05. AG + AA genotype and GG The 95% confidence interval is 1.15~3.74. The linkage disequilibrium between SNPs within the gene is very common. Conclusion The 267G/A polymorphism of megsin gene is associated with the susceptibility of IgA nephropathy, suggesting that AG and AA genotypes are the risk factors of IgA nephropathy.

Objective to observe the effect of overexpression of hyperpolarization-activated cyclic nucleotide-gated cation channel gene subtype (HCN) 4 on the electrophysiological characteristics of neonatal rat cardiomyocytes. Methods Twenty neonatal rat primary cardiomyocytes were isolated by enzymatic hydrolysis and cultured, and the cardiomyocytes were infected with recombinant adenovirus AdHCN4 containing HCN4 gene.The uninfected cells were used as the control group.The expression of HCN4 gene at mRNA and protein levels in cardiomyocytes were detected by RT-PCR and cell immunofluorescence method.The action potential and pacing current If of cardiomyocytes transfected with HCN4 gene were measured by whole cell patch clamp method.Results The expression levels of HCN4 mRNA and protein in cardiomyocytes of AdHCN4 infected group were significantly higher than those of control group; The frequency of spontaneous pulsation of myocardial cells in the infected group was (92.5±7.4) beats/min, which was significantly higher than that in the control group (68.9±6.2) beats/min (P<0.05); The action potential showed obvious 4-phase automatic depolarization, and its maximum diastolic potential (52.8±4.2) mV was significantly higher than that of control cells (-62.1±2.6) mV (P<0.05); There was no statistically significant difference between 50% and 90% time of action potential repolarization. The current density of pacing current If was (115.7±7.8) pA/pF in infected group cells, which was also significantly higher than that of control group cells (7.2±0.6) pA/pF (P<0.05). Conclusion Overexpression of pacing channel gene HCN4 can significantly enhance the autonomy of neonatal rat cardiomyocytes, thus increasing the frequency of spontaneous pulsation of cardiomyocytes, which suggests that HCN4 gene may be the target gene for the treatment of bradyarrhythmia.

Objective to investigate the effect of endogenous hydrogen sulfide (H2S) on nitric oxide (NO) /nitric oxide synthase (NOS) system in left-to-right shunt rats. Methods Male SD rats were randomly divided into shunt group, shunt + propargyl glycine (PPG) group, sham operation group and sham operation group + PPG group, with 8 rats in each group.Rats in shunt group and shunt + PPG group were established by abdominal aorta-inferior vena cava puncture to establish a left-to-right shunt animal model. Plasma NO, lung tissue H2S, NO content and NOS activity were measured; The contents of eNOS in lung tissues were determined by Western blot method. Results 4 weeks after shunt operation, there was no significant change in MPAP in shunt group compared with sham operation group; Plasma NO (23.2 μ mol/L ± 3.6 μ mol/L vs 17.9 μ mol/L ± 3.4 μ mol/L, P<0.05), lung tissue H2S (37.6 μ mol/mg ± 2.1 μ mol/mg vs 14.4 μ mol/mg ± 1.8 μ mol/mg, P<0.05), lung tissue NO (38.5 ± 6.5 μ mol/μ g vs 31.8 ± 6.5 μ mol/μ g, P<0.05) were compared between shunt group and sham operation group The activity of NOS in lung tissue (15.1 U/mg ± 2.4 U/mg vs 12.0 U/mg ± 1.4 U/mg, P<0.05) and the content of eNOS in lung tissue (0.3 ± 0.1 vs 0.2 ± 0.1, P<0.05) were significantly increased. 20.55% (19.5 mmHg±1.7 mm Hg vs 16.4 mmHg±1.7 mmHg and 19.5 mmHg±1.7 mmHg vs 15.5 mmHg±1.3 mmHg, P<0.05), H2S content in lung tissue decreased (28.8 μ mol/mg ± 2.2 μ mol/mg vs 37.6 μ mol/mg ± 2.1 μ mol/mg, P<0.05), while plasma NO (27.8 μ mol/L ± 4.8 μ mol/L vs 23.2 μ mol/L ± 3.6 μ mol/L, P<0.05), lung tissue NO (46.0 μ mol/μ g ± 6.0 μ mol/μ g vs 38.5 μ mol/μ g ± 6.5 μ mol/μ g, P<0.05), NOS activity (20.9 U/mg ± 3.9 U/mg vs 15.1 U/mg ± 2.4 U/mg, P<0.05) Conclusion Endogenous H2S may regulate pulmonary artery pressure in left-right shunt rats by inhibiting NO/NOS system.

Objective to construct a small hairpin structure RNA (shRNA) eukaryotic expression vector pEGFP-siRNA/TLR4 targeting Toll-like receptor (TLR) 4 mRNA, and to detect the inhibitory effect of RNA interference (RNAi) induced by this shRNA on the secretion of inflammatory factors stimulated by lipopolysaccharide in RAW264.7 cells, so as to investigate the inhibitory effect of RNAi targeting TLR4 gene on the inflammatory response of RAW264.7 cells. Methods A plasmid pEGFP-H1/siRNA carrying enhanced green fluorescent protein (EGFP) and shRNA cloning site was constructed, and the oligonucleotide fragment targeting TLR4 mRNA was designed by siRNA Wizard (http: / /www.simawizard.com/). The plasmid was cloned into pEGFP-H1/siRNA, and the eukaryotic expression plasmid pEGFP-H1/TLR4-siRNA expressing EGFP and TLR4-shRNA was constructed. The pEGFP-H1/TLR4-siRNA was transfected into cultured mouse macrophage line RAW264.7 by liposome transfection technology, and the cell line was observed RAW264.7 cells were stimulated with lipopolysaccharide, and the levels of inflammatory factors secreted by this cell line were detected by ELISA. Results the plasmids pEGFP-H1/siRNA and pEGFP-H1/TLR4-siRNA were digested by Bbs Ⅰ and MLU Ⅰ respectively. The bands of 4.9 kb and 340 bp appeared in the former, and the bands of 5.0 kb and 220 bp appeared in the latter. The length of the plasmid was consistent with the experimental design, and the sequence was confirmed to be correct. After transfection of pEGFP-H1/TLR4-siRNA into RAW264.7 cells, the expression rate of EGFP was 50% ± 8%. After stimulation with lipopolysaccharide, the levels of TNF-α in the cell culture medium of TLR4-siRNA transfected group (2 h: 825 pg/ml ± 136 pg/ml; 8 h: 2190 pg/ml ± 359 pg/ml) were significantly lower than those of control siRNA transfected group (2 h: 1179 pg/ml ± 240 pg/ml; 8 h: 4720 pg/ml ± 227 pg/ml, both P<0.01) Conclusion the shRNA targeting TLR4 mRNA may significantly inhibit the secretion of inflammatory factors induced by lipopolysaccharide in RAW264.7 cells through RNAi mechanism.

Objective to investigate the necessity of mitochondrial DNA A1555G mutation gene screening in high-risk and specific populations in the prevention of drug-induced deafness.Methods 1836 cases of sporadic non-syndromic deafness from different provinces and cities in China were screened for mitochondrial DNA A1555G mutation gene by self-developed mitochondrial DNA A1555G mutation detection kit.The positive individuals were screened out, and the status of all maternal family members of the positive cases was further understood, detailed family genealogy was drawn, and deafness prevention education was carried out for those without disease in the maternal lineage.Results Among 1836 cases, 63 cases had mitochondrial DNA A1555G mutation, the mutation rate was 3.43%; Among 63 maternal inheritance families, 8 cases were not followed up, and 3 cases were unwilling to provide family data; Among 52 families with complete follow-up data, 737 surviving maternal family members, 201 deafness (including probands) and 536 non-deafness. Conclusion It is an effective measure to prevent drug-induced deafness and reduce the incidence of drug-induced deafness by screening mitochondrial DNA1555G mutation in high-risk groups and specific groups.

Objective to investigate the correlation between single nucleotide deletion/insertion (4G/5G) polymorphism of plasminogen activator inhibitor-1 (PAI-1) gene promoter region-675 and the occurrence of PTE.Methods A case-control study was conducted. The case group consisted of 101 patients with PTE diagnosed by radionuclide pulmonary ventilation/perfusion imaging, spiral CT pulmonary angiography (CTPA) and clinical data. The genome of PAI-1 was extracted by potassium iodide-chloroform-isopropanol method, and the 4G/5G polymorphism in the promoter region of PAI-1 gene was analyzed by polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) and DNA direct sequencing. Results the frequencies of 4G and 5G alleles of PAI-1 gene were 0.495 and 0.505 in the control group, respectively, and the genotype distribution was in accordance with Hardy-Weinberg equilibrium law. In the control group, the frequencies of 4G alleles were 0.733 and 0.495,and the frequencies of 5G alleles were 0.267,0.505, respectively, χ 2=24.060, respectively. The relationship between 4G/5G and PTE was studied by univariate Logistic regression. The OR values of 4G4G +4G5G, 4G4G and 4G5G were 3.794 (1.786-8.060), 5.443 (2.416-12.260) and 2.450 (1.067-5.623), respectively. The P values were 0.001, 0.000 and 0.035, respectively. Conclusion 4G/5G and 4G/4G genotypes in PAI-1 promoter region were related to the occurrence of PTE, and the 4G/5G and 4G/4G genotypes showed higher risk of PTE.

Objective to compare the results of multidimensional pedicle fluoroscopy during operation, explore a simple and easy fluoroscopic angle, and reduce the misplacement rate of pedicle screws during operation. Methods Eight adult fresh L1-5 vertebrae specimens were taken, and three models were randomly established: medial, lateral and centered. The guide needle and pedicle screw were placed respectively, and the lateral position, orthogonal position, axial position of pedicle, axial position of guide needle and axial position of pedicle screw were sequentially fluoroscoped. The position of guide needle and pedicle screw was judged according to the relationship between the position of guide needle and pedicle screw and pedicle projection. The pedicle screw tract was scanned after operation to observe the medial and lateral cortical penetration of pedicle. The direct observation results of laminectomy were used as the gold standard. Results The accuracy of orthogonal fluoroscopy was 34%. The lateral fluoroscopy could not judge the error of medial and lateral deviation. The axial fluoroscopy of pedicle mistakenly judged the center of 4 pedicle screws as medial deviation, with an accuracy of 95%. The accuracy of axial fluoroscopy of guide needle is 100%. Conclusion The axial fluoroscopy of guide needle can accurately judge the position of pedicle screw guide needle, which can improve the accuracy of screw placement, Human factors in reducing pedicle screw loosening.

Objective to analyze the recurrence characteristics of T1G3 transitional cell carcinoma of bladder. Methods Among the recurrent transitional cell carcinoma of bladder in Peking Union Medical College Hospital from 1993 to 2003, 72 cases of T1 stage with the same first treatment method were selected and divided according to pathological grade, 37 cases in G3 group and 35 cases in G1-2 group. The data of first recurrence time, recurrence times, number, morphology, pathological stage of recurrent tumors and treatment methods after recurrence were analyzed respectively. Results The recurrence rates of T1G3 group at the first, second, third and fourth years were 29.7%, 40.5%, 18.9% and 10.8%, respectively, and T1G1-2 group were 14.3%, 34.3%, 31.4% and 20.0%, respectively. The difference between the two groups was statistically significant (P =0.025). 4 cases (10.8%) had single recurrence, 20 cases (54.0%) had more than three recurrences, and 17 cases (48.6%) and 6 cases (17.2%) in T1G1-2 group, respectively. The difference between the two groups was statistically significant (P<0.01). Moreover, the recurrence frequency is high, and most of them develop into invasive carcinoma after recurrence, so we should pay attention to clinical treatment and follow-up after operation.

Objective to investigate the effect and application prospect of diffusion tensor white matter bundle imaging (DTI) as a guide for minimally invasive neurosurgery for brain tumors. Methods 52 patients with brain malignant tumors were scanned. According to the fusion images, 34 patients underwent navigated minimally invasive tumor resection, and 18 patients underwent conventional minimally invasive craniotomy for brain tumor resection. The results, complications and follow-up data were analyzed. Results the secondary surgery rate was 0, the total tumor resection rate was 86.5% (45/52), the morbidity rate was 11.5% (6/52), and 1 case (1.9%) died. Conclusion the application of diffusion tensor white matter bundle imaging as a guide for malignant brain tumor surgery can promote the development of minimally invasive neurosurgery from cortical protection to nucleus protection and nerve bundle protection.

The key of successful brachial plexus block lies in accurate anatomical localization and uniform diffusion of local anesthetics. Recently, it has been reported abroad [1] that using ultrasound technique to guide brachial plexus block can further improve the effect of brachial plexus block. We observed the clinical effect of ultrasound-guided subclavian brachial plexus block with coracoid pathway and compared it with neurostimulator.

Oxaliplatin (oxalate platinum, L-OHP) is the third generation platinum anticancer drug after cisplatin and carboplatin. It has better efficacy, lower toxicity, improved pharmaceutical properties, expanded anticancer spectrum, and has no cross-resistance with cisplatin and carboplatin. The results of phase II clinical studies show that the efficacy of oxaliplatin alone in the treatment of newly treated colorectal cancer can reach 22%, which is close to that of 5-fluorouracil (5-FU) and calcium leucovorin (CF). The efficacy of oxaliplatin combined with 5-FU and CF can reach 30% ~40%, which has become the standard regimen of adjuvant chemotherapy for colorectal cancer and the first-line chemotherapy regimen for recurrent and metastatic colorectal cancer [1,2]. In addition, oxaliplatin has been widely used in the treatment of lung cancer, ovarian cancer and gastric cancer. However, like many other drugs, its toxic and side effects often limit the application of some patients, and the long-term toxic effects are particularly worrying. With the continuous deepening of the application research of oxaliplatin, the research on its neurotoxicity manifestations, mechanism and prevention and treatment methods have begun to take shape. Contributes to the continuity of the drug treatment course, so as to achieve better curative effect.

The use of experimental animal models to realistically simulate three-dimensional deformity of scoliosis IS the basis of the etiological research of idiopathic scoliosis (IS). From blind drug teratogenesis to pineal gland removal of chicken scoliosis animal models, the continuous progress of experimental scoliosis research has promoted the gradual deepening of the understanding of this disease. After more than half a century of exploration, experimental scoliosis animal models have involved many aspects of etiology such as genetics, central nervous system, muscle system, endocrine system and skeletal development. According to the preparation methods of animal models, we make a classification and review of experimental scoliosis.

Implantation of cardioverter-defibrillator (ICD) can terminate fatal ventricular tachycardia and ventricular fibrillation in vivo. There is a hypothesis that if there is no timely discharge of ICD, too long ventricular tachycardia will definitely lead to sudden cardiac death (SCD) in patients with non-ischemic cardiomyopathy. Therefore, in some previous small clinical trials on ICD installation in patients with non-ischemic cardiomyopathy, defibrillator-issued electric shock is considered as an alternative endpoint of sudden cardiac death. Although there is controversy about this, there has been a lack of effective evaluation methods. With the improvement of ICD technology, we can obtain electrocardiogram data at the time of electric shock, so as to judge whether ICD-issued electric shock is appropriate. Ellenbogen et al. of Virginia Medical University in the United States<Defibrillators in Non-Ischemic Cardiomyopathy Treatment Evaluation (DEFINITE) published in Circulation (2006, 113: 776-782) shows that shocks emitted by embedded cardioverter Defibrillators in patients with Non-Ischemic Cardiomyopathy cannot simply be considered sudden cardiac death.

Cerebral stroke occurs in about 1 million people in EU countries every year, of which ischemic stroke accounts for 80%, cerebral hemorrhage accounts for 15%, subarachnoid hemorrhage accounts for 5%. Among them, the incidence of stroke after the first year of transient ischemic attack (TIA) is about 12%. Therefore, anticoagulants and anti-platelet aggregation drugs are used to prevent stroke.

Some patients with atypical amniotic fluid embolism may only have postpartum hemorrhage as the main manifestation.Therefore,not all patients with amniotic fluid embolism show sudden circulatory failure and death.It can be a group of diseases from subclinical manifestations to rapid death.This suggests that some patients with amniotic fluid embolism with postpartum hemorrhage as the main manifestation may be misdiagnosed or undiagnosed,so they cannot be treated in time.This study preliminarily discussed the relationship between STN antigen and postpartum hemorrhage by measuring the concentration of Sialyl Tn (STN) antigen in maternal serum 2 h after deliver,and preliminarily discussed the risk of atypical amniotic fluid embolism or amniotic fluid embolism with postpartum hemorrhage as the main manifestation in postpartum hemorrhage population.It provides a theoretical basis for more effective clinical treatment of some patients with postpartum hemorrhage.

Both cardiopulmonary bypass and myocardial ischemia-reperfusion can lead to the production of a large number of inflammatory factors in the whole body and the local myocardium. At present, aprotinin is added to the prefilled solution to inhibit the inflammatory response during cardiopulmonary bypass and myocardial ischemia-reperfusion, but the study of adding aprotinin directly to the perfusion has not been reported. Therefore, we directly added aprotinin to the perfusion solution to observe the changes of coronary sinus endothelin-1 (ET-1), interleukin-8 (IL-8), troponin Ⅰ (cTnI) and adenosine triphosphate (ATP) levels at various time points before and after arrest, and the effect of aprotinin on the above indexes, so as to explore the myocardial protective effect of perfusion containing aprotinin.

The blood-brain barrier is composed of cerebral microvascular endothelial cells, astrocyte (As) foot processes, basement membrane and perithelial cells, which are the main structural basis of BBB, and As plays an important role in maintaining the integrity of BBB [1]. In this study, human umbilical vein endothelial cells (HUVEC) were co-cultured with mouse As to construct a model of BBB in vitro, and its morphology and barrier function were identified, so As to become a simple and reliable tool for studying the permeability of substances (including drugs) through BBB in vitro.

Intertrochanteric fractures tend to occur in the elderly, and improper treatment often leaves coxa varus, lower limb shortening and external rotation deformity.Studies have shown that the probability of femoral trochanteric fractures in the elderly is significantly higher than that of the femoral neck [1]. At present, there are many treatment methods for intertrochanteric fractures of the femur, and it is more suitable for elderly patients to choose less trauma, strong fixation and early movement.Percutaneous plate internal fixation technology is widely used in long bone fractures and plateau fractures of the extremities, but there are few articles reporting intertrochanteric fractures of the femur. We used closed reduction, modified small incision and DHS minimally invasive implantation to achieve good results, which are reported as follows.

A 41-year-old male was admitted to hospital for 1 month due to chest tightness and palpitations. The orthopedic X-ray of chest showed a localized protrusion of the right heart margin; Cardiac ultrasound showed that there was an elliptical solid hypoechoic mass attached to the atrial wall near the atrial septum of the right atrium, with a size of about 3.5 cm ×3.1 cm ×3.5 cm. The surrounding echo was enhanced and seemed to have a capsule. Color Doppler flow imaging (CDFI) showed that there was no blood flow signal in the mass. The preoperative diagnosis was right atrial tumor. Cardiac tumor resection was performed under general anesthesia and cardiopulmonary bypass.