Volume 120  ·   Issue 22  ·  2007

The practice of modern anaesthesiology has extended into perioperative medicine. Due to their expertise in analgesic drug pharmacology and peripheral nerve blocking, anaesthesiologists have pioneered in the management of acute postoperative pain. Effective postoperative analgesia reduces the incidence of postoperative chronic pain, improves the functioning of organs following surgery and shortens the hospital stay.1,2 Although a variety of analgesic techniques and preventative approaches are at the disposal of modem aneasthesiologists, including patient controlled epidural analgesia (PCEA), patient controlled intravenous analgesia, multimodal analgesia and pre-empty analgesia.Despite this array of strategies, these predominantly opioid based techniques are still limited by side-effects such as vomiting, nausea, itching and urinary retention.To optimize further the management of acute postoperative pain, basic mechanisms of postoperative pain must be explored and new treatments must continue to be developed。

In the recent editorial comment, Duboule1 emphasized that "animal development is, in fact, nothing but time".That a circadian timing system is apparently universal in biology is the evidence for the important physiological role that rhythmicity plays。

Abstract：advanced glycation end products; comet assayBackground The accumulation of free radicals and advanced glycation end products (AGEs) in cell plays a very important role in replicative senescence. Aminoguanidine (AG) has potential antioxidant effects and decreases AGE levels. This study aimed to investigate its effect on replicative senescence in vitro.Methods The effects of aminoguanidine on morphology, replicative lifespan, cell growth and proliferation, AGEs, DNA damage, DNA repair ability and telomere length were observed in human fetal lung diploid fibroblasts (2BS).Results Aminoguanidine maintained the non-senescent phenotype of 2BS cells even at late population doubling (PD) and increased cumulative population doublings by at least 17-21 PDs. Aminoguanidine also improved the potentials of growth and proliferation of 2BS cells as detected by the MTT assay. The AGE levels of late PD cells grown from early PD in DMEM containing aminiguanidine decreased significantly compared with those of late PD control cells and were similar to those of young control cells. In addition, the cells pretreated with aminoguanidine had a significant reduction in DNA strand breaks when they were exposed to 200 μmol/L H2O2 for 5 minutes which indicated that the compound had a strong potential to protect genomic DNA against oxidative stress. And most of the cells exposed to 100 μmol/L H2O2 had much shorter comet tails and smaller tail areas after incubation with aminoguanidine-supplemented DMEM, which indicated that the compound strongly improved the DNA repair abilities of 2BS cells. Moreover, PD55 cells grown from PD28 in 2 mmol/L or 4 mmol/L aminoguanidine retain telomere lengths of 7.94 kb or 8.12 kb, which was 0.83 kb or 1.11kb longer than that of the control cells.Conclusion Aminoguanidine delays replicative senescence of 2BS cells and the senescence-delaying effect of aminoguanidine appear to be due to its many biological properties including its potential for proliferation improvement, its inhibitory effect of AGE formation, antioxidant effect, improvement of DNA repair ability and the slowdown of telomere shortening。

Abstract：Background Most cardiac regenerative approaches can restore injured heart muscles. In this study, we investigated if fibrin sealant could help neonatal cardiomyocytes restore myocardial function in a rat model of myocardial infarction.Methods The left anterior descending artery in adult female Sprague-Dawley (SD) rats was ligated to make a myocardial infarction model. Neonatal ventricular cardiomyocytes from one-day male SD rats were isolated, labeled and cultured. The cells were injected into the infarcted area three weeks later. The animals were randomized into four recipient groups: (1) cardiomyocytes plus fibrin sealant (group CF, n=10); (2) cardiomyocytes alone (group C, n=10); (3)fibrin sealant recipients alone (group F, n=10); (4) control group (n=10). Four weeks after transplantation,echocardiography and Langerdoff model were used to assess heart function. Immunohistochemical staining and polymerase chain reaction (PCR) were performed to track the implanted cardiomyocytes and detect the sex-determining region Y gene on Y chromosome.Results Echocardiography showed the fraction shortening (FS) in groups CF, C, F and control group was (27.80±6.32)%, (22.29±4.54)%, (19.24±6.29)% and (20.36±3.29)% respectively with statistically significant differences in group CF compared with the other groups (P＜0.05). The Langendoff model revealed that the left ventricular development of peak pressure (LVDPmax, mmHg) in groups CF, C, F and control group was 104.81±17.05, 80.97±21.60, 72.07±26.17 and 71.42±17.55 respectively with statistically significant differences in group CF compared with the other groups (P＜0.05). Pathological examination and PCR indicated that transplanted cardiomyocytes in group CF survived better than those in the other groups.Conclusion Transplanted neonatal cardiomyocytes plus fibrin sealant can survive in myocardial infarctioned area and improve heart function greatly in rat models。

Abstract：Background Serum osteoprotegerin (OPG) and matrix metalloproteinase-2 (MMP-2) have been shown to play a role in bone metabolism by degrading the bone matrix. The present study was undertaken to compare OPG and MMP-2 with bone mineral density and three markers (alkaline phosphatase (AKP), calcium and phosphorus) in postmenopausal women in Wuhan.Methods Serum OPG, MMP-2, and AKP of 78 Chinese postmenopausal women aged 48 to 65 were measured using enzyme-linked immunosorbent assay (ELISA). Bone mineral density was measured with dual energy X-ray absorptiometry (DEXA), and serum calcium and phosphorus were measured by auto biochemical analysis.Results Serum OPG and MMP-2 concentrations were significantly higher in postmenopausal women with osteoporosis ((127.6±6.3) ng/L; (1388±121) μg/L)) than those in age-matched normal controls ((72.3±2.4) ng/L; (1126±141) μg/L,P＜0.01). Negative relationships were found between serum OPG, MMP-2 levels and bone mineral density in osteoporotic women. Adjusted by age and body mass index (BMI), the correlation of MMP-2 with bone mineral density of the neck of the femur disappeared. In osteoporotic women, negative correlations between OPG, MMP-2 levels and serum calcium were found (r=-0.216; r=-0.269, P＜0.05), but positive correlations between OPG and serum AKP, serum phosphorus (r=0.235; r=0.124, P＜0.05).Conclusions Significant correlations exist between serum OPG, MMP-2 levels and bone metabolism in high bone turnover of postmenopausal osteoporotic women. The concentrations of serum OPG and MMP-2 increase possibly as a concomitant event in the high bone turnover state, such as postmenopausal osteoporosis. Therefore serum OPG and MMP-2 could be used as indicators for the bone metabolism in postmenopausal osteoporotic women。

Abstract：Background The large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies.Methods scFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC Ⅲ encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated.Results The phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression.Conclusions The reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy。

Abstract：Background Human immunodeficiency virus (HIV-1) is divided into two types, HIV-1 (groups M, N and O) and HIV-2.Heilongjiang Province located in the northeast of China, and the feature of the subtype distribution and sequence characteristics of HIV-1 strains prevalent in Heilongjiang Province is still uncertain. The aim of this study was to investigate the subtype distribution and genetic characteristics of HIV-1 strains in one hospital in Heilongjiang Province.Methods HIV-1 env gene was amplified by nested-PCR from peripheral blood mononuclear cells (PBMCs) obtained from 19 HIV-1 seropositive individuals in Heilongjiang Province. The C2-V3 region was sequenced. Aligned the nucleotide sequence of 19 samples with CLUSTAL W (BioEdit) software, results were acquired and used for phylogenetic tree analysis after artificial adjustment. Reference sequence, downloaded from Los Alamos HIV Sequence Database,was used to identify the subtype of obtained sequence. Genetic distance between sequences was assessed using the software MEGA 3.1 Kimura 2-parameter, and the Phylogenetic tree was reestablished with Neighbor-Joining method.Results Phylogenetic tree analysis showed that 19 Heilongjiang strains clustered closely to subtype B strain from Thailand and were far from other international subtype reference strains. Statistical test showed no significant discrepancy between the genetic distance of interclass and intra-class (P＞0.05). The analysis of V3 loop amino sequence of 19 Heilongjiang B strains revealed that V3 tip motif of 10 samples (52.63%) was GPGQ, and of 4 samples (21.53%) was GPGR.Conclusions The subtype of 19 HIV-1 seropositive individuals in Heilongjiang Province is B', and it is introduced from He'nan Province. V3 tip motifs of the HIV-1 isolates are mainly GPGQ and GPGR。

Abstract：Background There has been continuous debate as to whether Y chromosome loss is an age related phenomenon or a cytogenetic marker indicating a malignant change. This study aimed to investigate the frequency of Y chromosome loss in the specific patients in order to determine whether it is an age related phenomena or a cytogenetic marker indicating a malignant change.Methods Five hundred and ninety-two male patients with a median age of 59 years old (22-95 years) were included in this study. These patients were divided into two groups: the study group, including 237 patients who had hematological disorders included myeloproliferative disorder (MPD), myelodysplastic syndrome (MDS), acute myeloid leukemia (AML),chronic myeloid leukemia (CML), multiple myeloma (MM), and lymphoma and the control group including 355 patients with no evidence of hematological disease. Both conventional cytogenetics and fluorescence in situ hybridization using DNA probes specific for the centromere of chromosomes X or Y were performed according to our standard laboratory protocols.Results Twenty-four out of 237 patients with hematological disorders (10.1%) had Y chromosome loss. Of these 24patients, 2 patients had AML (5.0% of all AML patients), 2 patients had CML (5.7% of all CML patients), 2 patients had MPD (8.0% of all MPD patients), 3 patients had MM (10.0% of all MM patients), 5 patients had lymphoma (10.6% of all lymphoma patients) and 10 patients had MDS (16.7% of all MDS patients). Twenty-one out of these 24 patients had a loss of Y chromosome as the sole anomaly and the remaining three had a loss of Y chromosome accompanied with otherstructural changes detected by conventional cytogenetic analysis. Fluorescence in situ hybridization (FISH) analysis confirmed the routine cytogenetic results. All 24 patients had a loss of Y chromosome with a range of 17.5%-98.5% of cells. Two of the patients, one with AML and another with CML, had karyotype and FISH testing done both at the initial diagnosis and during remission. The results showed a loss of Y chromosome at initial diagnosis but a normal 46,XY karyotype during remission. Only 9 out of 355 patients (2.5%) without evidence of hematological disease had Y chromosome loss, among them 7 patients had cardiovascular diseases and 2 patients had kidney diseases. Comparison of the incidence of Y chromosome loss in patients with hematological disorders or without evidence of hematological disease using statistical analysis showed a statistically significance difference (P＜0.05).Conclusions The present study demonstrated that the frequency of Y chromosome loss is significantly higher in patients with hematological disorders than in patients without hematological disorders, which indicates that the loss of Y chromosome is associated with a neoplastic change。

Abstract：Background Previous studies have shown that oncosis in malignant tumors might be related to cellular energy supply.The aim of this study was to detect oncosis in human esophageal squamous cell carcinoma (ESCC), and to investigate its relationship with apoptosis and microvessel density (MVD).Methods ESCC specimens were obtained from 30 patients with ESCC after surgery. Transmission electron microscopy,TUNEL, and immunohistochemistry were used to detect oncosis, apoptosis, and MVD. The relation of oncosis to apoptosis and MVD was analyzed by ANOVA, t test, and q test using SPSS 10.0.Results Transmission electron microscopy revealed both oncosis and apoptosis in the ECSS tissues. About 10% of the TUNEL-positive cells, which were considered apoptotic cells, showed the characteristics of oncosis. In the areas, where oncotic cells accumulated, apoptotic cells were rare; contrarily, where apoptotic cells gathered, oncotic cells were sparse.Compared with the tissues with a high MVD, the number of oncotic cells was increased and that of apoptotic cells was decreased in the tissues with a low MVD.Conclusions Cellular oncosis can be detected in human ESCC tissues. The distribution of oncotic cells presents a close relationship with cellular apoptosis and MVD. Oncosis might be induced by poor blood supply。

Abstract：Background Melilotus suaveolens Ledeb (M. suaveolens Ledeb) has long been used as a folk medicine in inflammation-related therapy. This study was undertaken to determine the anti-inflammatory effect of the plant.Methods Petroleum ether fraction, ethyl acetate fraction, n-butanol fraction, aqueous fraction were obtained from ethanol extract of M. suaveolens Ledeb and evaluated by high performance liquid chromatography (HPLC). While dexamethasone (DM) was used as a positive control, the effects of different solution fractions of ethanol extract on tumor necrosis factor α (TNF-α) mRNA, cyclooxygenase 2 (COX-2) mRNA, COX-2 and nuclear factor κB (NF-κB) of LPS-stimulated RAW 264.7cells were studied by real-time PCR, Western blot analysis and immunocytochemical assay, respectively.Results Coumarin was one of the main ingredients in different solution fractions of ethanol extract except the aqueous fraction with no inflammatory effect. The petroleum ether fraction, ethyl acetate fraction and n-butanol fraction of ethanol extract could inhibit the production of TNF-α mRNA, COX-2 mRNA and NF-κB to some extent.Conclusions Different solution fractions of ethanol extract from M. suaveolens Ledeb had similar anti-inflammatory effect as did dexamethasone except the aqueous fraction. Coumarin was likely to be essential to the anti-inflammatory effect, and other ingredients might attribute to their different anti-inflammatory effects from the HPLC fingerprint。

Abstract：Background Early local platelet activation after coronary intervention identifies patients at increased risk of acute stent thrombosis (AST). However, early changes in platelet activation in coronary circulation following drug-eluting stent (DES) implantation have never been reported.Methods In a prospective study of 26 consecutive elective stable angina patients, platelet activation was analyzed by measuring soluble glycoprotein V (sGPV) and P-selectin (CD62P) before and after implantation of either DES or bare metal stent (BMS). All patients were pretreated with clopidogrel (300 mg loading dose) and aspirin (75 mg orally) the day before the procedure. Blood samples were drawn from the coronary ostium and 10 - 20 mm distal to the lesion site.Results Consistent with the lower baseline clinical risk, the levels of CD62P and sGPV were within normal reference range, both in the coronary ostium and distal to the lesion before percutaneous coronary intervention (PCl) procedure.The levels of CD62P and sGPV did not change significantly (CD62P: (31.1 ± 9.86) ng/ml vs (29.5 ± 9.02) ng/ml, P=0.319and sGPV: (52.4 ± 13.5) ng/ml vs (51.8 ± 11.7) ng/ml, P=0.674, respectively) after stent implantation when compared with baseline. Changes in these platelet activation markers did not differ between stent types.Conclusions Intracoronary local platelet activation does not occur in stable angina patients before and immediately following DES implantation when dual anti-platelet is administered。

Abstract：Background Transmyocardial laser revascularization (TMLR) has been used in the treatment of patients with end-stage coronary artery disease (CAD) since 1990. The aim of this study was to evaluate the long-term effectiveness of TMLR in patients with diffuse CAD.Methods Ninety-four consecutive patients underwent TMLR in one center from July 1997 to December 2000. The follow-up data of these patients were obtained through face-to-face, mail questionnaires, or telephone interviews in July 2004 and December 2004. Four cases failed to respond. Mean follow-up time was (5.5±1.0) years.Results Mean Canadian Cardiovascular Society (CCS) angina scores of TMLR patients were 3.1±0.8 at baseline,1.7±0.9 at 1 year (P＜0.05), 1.7±0.9 at 3 years (P＜0.05), and 1.9±0.9 at 5 years (P＜0.05). At an average of 5 years, 69%of the patients had ≥ 1 angina class reduction, mean NYHA class level (1.9±0.9) ameliorated compared to the baseline (2.5±0.7, P＜0.001), the rate of re-hospitalization was 2.7 times/person. Kaplan-Meier survival rate was 87% at 1 year,69% at 3 years and 64% at 5 years. The causes of death were attributed more to heart failure (58.9%) and myocardial infraction (14.7%) after TMLR. The patients with no angina relief, or who died after TMLR, had a higher percentage of preoperative unstable anginas or prior myocardial infraction compared to the survivors. The assorted shapes of myocardial laser channels were detected in some patients by the color Doppler velocity technique.Conclusions TMLR provided a long-term improvement in the quality of life, including CCS angina class or NYHA heart functional class for about 70% of Chinese patients with severely disabling angina pectoris. The various myocardial laser channels would always be visible after TMLR. 5-years after TMLR as a sole therapy, the survival rate of the patients was 64%。

Atrial fibrillation (AF) is the most common cardiac arrhythmia in man, and epidemiological study has shown the close relationship between the incidence of AF and aging.1 Although the effects of aging on atrial electrophysiology have been investigated in humans and animal models,2-9 the electrophysiological changes that make the atria of aged individuals more susceptible to AF than those of adults remain poorly understood。

The action potential of pacemaker cells in the sino-atrial node forms the automatic rhythm of the heart. The automatic depolarization in phase 4 is the basis of the automaticity in pacemaker cells. Many currents are included in phase 4, such as calcium current,1TTX-sensitive sodium current,2 sustained inward current (Isi),3 decay of delayed rectifier potassium current,4 etc.Funny current (If) has long been recognized as important in this phase and is activated at hyperpolarized potentials during cell diastole and in turn activates other currents to form automatic depolarization.5 It is controlled by voltage and cAMP, an intracellular messenger, so the auto no-mic nerve can modulate this current by increasing or decreasing intracellular cAMP。

Catheter ablation of atrial fibrillation (AF) has been increased dramatically recently.1 However, it is an unpleasant procedure with intolerable pain without sedation. Propofol and fentanyl/midazolam have been widely used in painful clinical examination and cardiovascular procedures with established safety and efficacy.2,3 Propofol, alfentanyl and midazolam were administrated for catheter ablation in some electrophysiological labs for a less painful procedure.4However, there is few published work on the sedation regimen for catheter ablation of AF。

Von Hippel-Lindau (VHL) disease is a rare autosomal dominant disorder caused by germ line mutations of the VHL tumour suppressor gene. It predisposes affected individuals to develop a variety of neoplasms, including haemangioblastomas of the central nervous system,retinal angiomas, renal cell carcinomas (RCCs),pheochromocytomas and cysts of the kidneys and epididymis。

Diffuse panbronchiolitis (DPB) is a relatively recently recognized, idiopathic progressive disease, originally described in Japan in 1969.1 It has distinctive clinicopathological features characterized by chronic pulmonary inflammation in the respiratory bronchioles of the lung and paranasal sinuses.2 It has been reported more extensively among Japanese than other ethnic populations.The first case of DPB in China was reported in 1996.3Since then, an increasing number of cases have been described, which suggest that this disease may not be rare in China. However, clinicians are not familiar with this disease and it may be 4actually under-recognized and under-reported in China.4 Therefore we summarize the cases reported in the mainland of China since 1996,hoping that this would help to increase the awareness of this disease。

Spontaneous regression of pituitary macroadenomas is rare. It is generally thought that absorption of the necrotic tissue after pituitary apoplexy may cause the spontaneous healing of the tumor. However, such cases are often accompanied by hypopituitarism, which sould be treated with hormone replacement therapy. Here we report two cases of spontaneous remission of pituitary macroadenomas in women, who had complete tumor disappearance with preserved pituitary function。

Holmes' tremor has been postulated as a syndrome attributed to those lesions that interrupt the dentatethalamic and the nigrostriatal tracts thus causing both an action and a rest tremor.1 It may arise from various underlying structural disorders including multiple sclerosis, stroke, or tumors. So far, to our knowledge, few studies on Holmes' tremor secondary to cavernoma have been reported.2 Here we report a case of disabling tremor,who harbored a cavernoma in the midbrain。

Paroxysmal nocturnal hemoglobinuria (PNH) is a consequence of nonmalignant clonal expansion of one or several hematopoietic stem cells that have acquired a somatic mutation of phosphatedalinositol glycae-class A (PIGA) gene leading to deficient glycosyl phosphatidylinositol (GPI) synthesis。

Pulmonary vein varix, congenital or acquired dilation of the pulmonary vein, is a rare vascular abnormality.This disease often presents as a pulmonary mass on chest roentgenography, and pulmonary angiography, computed tomography (CT), or magnetic resonance imaging (MRI)is considered necessary for diagnosis.1 The diagnosis of the disease by thoracic exploration is rare because the present case was diagnosed as a pulmonary arteriovenous fistula before operation。